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 2012 , Vol 16 , Num 1
Synthesis and evaluation of cytotoxic activities of some 1,4-disubstituted thiosemicarbazides, 2,5-disubstituted-1,3,4-thiadiazoles and 1,2,4-triazole-5-thiones derived from benzilic acid hydrazide
Işıl Çoruh1, Sevim Rollas1, Suna Özbaş Turan2, Jülide Akbuğa2
1Marmara Üniversitesi, Eczacılık Fakültesi, Farmasötik Kimya, İstanbul, Türkiye
22Marmara Üniversitesi, Eczacılık Fakültesi, Farmasötik Biyoteknoloji, Istanbul, Türkiye
DOI : 10.12991/201216421


Bu çalışmada 1-(α,α-difenil-α-hidroksi)asetil-4-sübstitüetiyosemikarbazid (2a-k), [5-(sübstitüeamino) -1,3,4-tiyadiazol- 2-il] (difenil)metanol (3a-b) ve 3-[hidroksi(difenil)metil]-4-(nonsübstitüe/sübstitüe)-2,4-dihidro-5H-1,2,4-triazol-5- tiyon (4a-c) yapısındaki yeni bileşikler sentez edilmiş ve bileşiklerin sitotoksik etkinlikleri araştırılmıştır. Bu amaçla benzilik asit metil esterinin hidrazin hidrat ile etanollü ortamda reaksiyonu ile benzilik asit hidrazidi (1) elde edilmiş, 1'in uygun alkil/aril isotiyosiyanatlara katımı ile 1-(α,α-difenil-α-hidroksi)asetil-4-sübstitüetiyosemikarbazid (2a-k) kazanılmıştır. [5-(sübstitüeamino)-1,3,4-tiyadiazol-2-il](difenil)metanol türevi (3a-b) bileşikler, 2a ve 2c' nin derişik sülfürik asit ile siklizasyonu sonucu elde edilmiştir. 3-[hidroksi(difenil)metil]-4-(nonsübstitüe/sübstitüe)-2,4-dihidro-5H- 1,2,4-triazol-5-tiyon türevi (4a-c) bileşikler ise 2c, 2d ve 2g'nin 2N NaOH ile reaksiyonundan kazanılmıştır. Bileşiklerin yapıları elemental analiz, HPLC, UV, IR, 1H-NMR, 13C-NMR, HSQC ve MS verileri ile doğrulanmıştır. Bileşik 2a, 2c-k, 3b ve 4b HEK293 hücre hattı kullanılarak MTT yöntemiyle sitotoksik etkinlikleri araştırmak üzere seçilmiştir. Özellikle 1-(α,α-difenil-α-hidroksi)asetil-4-siklohekzilmetiltiyosemikarbazid (2e) bileşiğinde 10 mg/ml konsantrasyonda % 50.23 sitotoksik aktivite saptanmıştır.


The use of thiosemicarbazide in organic synthesis has become a classical strategy for the synthesis of several heterocycles. Among the increasing number of heterocyclic sulphur and nitrogen containing compounds, which are being pursued in both industry and academia, 1,3,4-thiadiazole and 1,2,4-triazole derivatives are also interesting targets for drug design. Therefore, there have been intense investigations on 1,4-disubstitutedthiosemicarbazide, 1,3,4-thiadiazole and 1,2,4-triazole- thione compounds.

1,4-Disubstituted-thiosemicarbazide are biologically versatile compounds displaying a variety of biological effects which include anti-inflammatory[1], antimycobacterial[2,3], antimicrobial[4-6], antifungal[7], antibacterial[8,9] and antiviral[3] activities. 2,5-Disubstituted 1,3,4-thiadiazoles, synthesis of which frequently include the reaction of acylthiosemicarbazides with acidic reagents such as concentrated sulfuric acid, possess various biological properties such as anticonvulsant[10], antifungal[11,12], antituberculosis[13-15], antimicrobial (16, 17), anti-inflammatory (18), cytotoxic[19], and antiproliferative[20], antioxidant[21-22] activities. In addition, 1,2,4-triazole-thiones which possess important pharmacological activities such as anticonvulsant[23], anti-inflammatory[24-28], antibacterial[29], cytotoxic[30,31], antimicrobial[32], anticancer (33,34), and antiviral[35] have found wide use in medicinal chemistry as common structures.

The present communication deals with the synthesis of 1-(α,α-diphenyl-α-hydroxy) acetyl-4- substitutedthiosemicarbazide (2a-k), [5-(substi- tutedamino) -1,3,4-thiadiazole-2-yl] (diphenyl) methanol (3ab) and 3-[hydroxy (diphenyl) methyl]-4-(nonsusbtituted/ substituted)-2,4-dihydro-5H-1,2,4-triazole-5-thione derivatives (4a-c). Their structures were confirmed by means of UV, IR, 1H-NMR, Mass spectral data and elemental analysis. All the synthesized compounds were screened for their cytotoxic activities by using HEK293 cell line of MTT assay.

All solvents and chemicals used in this study were supplied from Aldrich, Merck and Fluka and used without purification. Melting points (ºC) were measured using Schmelzpunktbestimmer SMP II melting point apparatus, uncorrected. The reactions were monitored on Merck pre-coated aluminium TLC plates 60F–254 and the products were visualized by UV-light using ethyl asetate and ether (50:50, v/v) as solvent system. The UV spectra were recorded on a Schimadzu UV-1601 spectrophotometer. The Infrared spectra were recorded on Schimadzu FTIR-8400S Spectrophotometer and expressed in wave number ν (cm-1). 1H-NMR spectra were recorded on Bruker AVANCE-DPX 400 and Varian Mercury Spectrometer at 400 MHz, using DMSO-d6 as a solvent; tetramethylsilane (TMS) was used as internal standard. All NMR chemical shifts are reported as δ values in parts per million (ppm) and coupling constant (J) are given in hertz (Hz). Mass spectra were obtained by using Agilent 1100 LC-MS and Waters 2695 Alliance Micromass. 1H-NMR and Mass analyses were provided by Faculty of Pharmacy Center Laboratory (Ankara University) and the Scientific and Technical Research Council of Turkey, TÜBİTAK (Ankara).

Preparation of benzilic acid hydrazide (1)
Hydrazine hydrate (0.03 mol, 80%) was added to benzilic acid methyl ester (0.01mol). The mixture was refluxed at 100°C for 30 minutes. After adding ethanol (10.0 mL), the mixture was heated in a water steam bath for two hour. The residue was filtered, washed with water and recrystallized from ethanol. mp: 171–172 °C, (lit. [36] mp. 169-170 °C).

General procedure for the synthesis of 1-(α,α-diphenyl-α- hydroxy)acetyl-4-(substituted)thiosemicarbazide (2a-k)

Equimolar amounts of benzilic acid hydrazide (0.01 mol) and appropriate alkyl/aryl isothiocyanates were refluxed in ethanol (30 mL) for 2-3 hours. The crystalline product was filtered and recrystallized from ethanol to obtain 1,4-disubstitutedthiosemicarbazides.

1-(α,α-diphenyl-α-hydroxy)acetyl-4-(2-chloroethyl)thiosemicarbazide (2a): White crystals, yield 86%, mp 260–262 oC; IR (νmax, cm-1): 3248 (NH), 1707 (C=O), 1244 (C=S). 1H NMR (400 MHz, DMSO-d6) δ (ppm): 3.51 (-CH2Cl, protons are shadowed with solvent), 3.97 (2H, t, NHCH2), 7.08 (1H, s, OH), 7.21–7.63 (10H, m, Ar-H), 10.80 (1H, N4-H), 11.09 (1H, s, N2-H), 12.00 (1H, s, N1-H). Anal. Calcd for C17H18ClN3O2S (363.86): C, 56.12; H, 4.99; N, 11.55; S, 8.81%. Found: C, 56.23; H, 4.85; N, 11.48; S, 8.99%. (API-ES+, m/z, %): 328 [M+-35.5] (100), 310, 282, 210, 209, 208, 178, 150, 143.

1-(α,α-diphenyl-α-hydroxy)acetyl-4-(4-cyanophenyl)thiosemicarbazide (2b): White crystals, yield 85%, mp 169–173 oC; IR (νmax, cm-1): 3363 and 3300 (NH), 1687 (C=O), 1282 (C=S). 1H NMR (400 MHz, DMSO-d6) δ (ppm): 6.82 (1H, s, OH), 7.24– 7.78 (14H, m, Ar-H), 9.34 (1H, N2-H), 10.12 (1H, s, N1-H), 10.50 (1H, s, N4-H). Anal. Calcd for C22H18N4O2S (402.46): C, 65.65; H, 4.51; N, 13.92; S, 7.97%. Found: C, 65.33; H, 4.34; N, 14.08; S, 8.38%. (API-ES+, m/z, %): 403 [M+], 388, 386, 385 (100), 379, 366, 355, 347, 342, 307, 301, 294, 268, 264, 254, 246.

1-(α,α-diphenyl-α-hydroxy)acetyl-4-benzoylthiosemicarbazide (2c): White crystals, yield 42%, mp 189–190 oC; IR (νmax, cm- 1): 3232 (NH), 1674 (C=O), 1251 (C=S). 1H NMR (400 MHz, DMSO- d6) δ (ppm): 7.28 (1H, s, OH), 7.13–8.04 (15H, m, Ar-H), 10.86 (1H, N4-H), 11.93 (1H, s, N2-H), 13.17 (1H, s, N1-H). Anal. Calcd for C22H19N3O3S (405.46): C, 65.17; H, 4.72; N, 10.36; S, 7.91%. Found: C, 64.91; H, 4.60; N, 10.36; S, 7.16%. (API-ES+, m/z, %): 407 (M++H)+, 389 (100), 388, 338, 276, 247, 229, 199, 157.

1-(α,α-diphenyl-α-hydroxy)acetyl-4-(4-trifluoromethoxyphenyl) thiosemicarbazide (2d): White crystals, yield 48%, mp 145–146 ºC; IR (νmax, cm-1): 3300 and 3142 (NH), 1653 (C=O), 1261 (C=S). 1H NMR (400 MHz, DMSO-d6) δ (ppm): 6.78 (1H, s, OH), 7.65–7.14 (12H, m, Ar-H), 9.28 (1H, N4-H), 9.97 (1H, s, N2-H), 10.50 (1H, s, N1-H). Anal. Calcd for C22H18F- 3N3O3S (461.45): C, 57.26; H, 3.93; N, 9.11; S, 6.95%. Found: C, 57.37; H, 4.00; N, 9.08; S, 6.63%. (API-CI+, m/z, %): 462 [M+], 446, 445, 444 (100), 252, 225, 223, 208, 79.

1-(α,α-diphenyl-α-hydroxy)acetyl-4-cyclohexylmethylthiosemicarbazide (2e): White crystals, yield 94%, mp 194–196 oC; IR (νmax, cm-1): 3162 (NH), 1687 (C=O), 1294 (C=S). 1H NMR (400 MHz, DMSO-d6) δ (ppm): 0.65–1.84 (11H, m,cyclohexyl), 3.28 (2H, m, –CH2), 6.76 (1H, s, OH), 7.09–7.59 (11H, m, Ar-H and N4-H), 9.34 (1H, s, N2-H), 10.26 (1H, s, N1-H). Anal. Calcd for C22H27N3O2S (397.53): C, 66.47; H, 6.85; N, 10.57; S, 8.07%. Found: C, 66.32; H, 6.84; N, 10.51; S, 8.04%. (API-ES+, m/z, %): 398 [M+], 381, 342, 304, 292, 264, 263, 225, 208, 188, 173.

1-(α,α-diphenyl-α-hydroxy)acetyl-4-(3-pyridyl)thiosemicarbazide (2f): White crystals, yield 85%, mp 184–186 ºC; IR (νmax, cm-1): 3182 (NH), 1651 (C=O), 1257 (C=S). 1H NMR (400 MHz, DMSO-d6) δ (ppm): 6.75 (1H, s, OH), 7.17–7.38 ve 7.45–7.60 (10H, m, Ar-H), 7.40 (1H, dd, J=8.07 Hz, 8.08 Hz, pyridine-H5), 7.93 (1H, d, pyridine-H4), 8.36 (1H, d, pyridine-H6), 8.53 (1H, s, pyridine-H2) 9.33 (1H, N4-H), 10.02 (1H, s, N2-H), 10.56 (1H, s, N1-H). Anal. Calcd for C20H18N4O2S (378.44): C, 63.47; H, 4.79; N, 14.80; S, 8.47%. Found: C, 63.26; H, 4.83; N, 14.58; S, 8.15%. (API-CI+, m/z, %): 379 [M+] (100), 243, 226, 225, 197, 139, 138, 137.

1-(α,α-diphenyl-α-hydroxy)acetyl-4-(3,5-bistrifluoromethylphenyl) thiosemicarbazide (2g): White crystals, yield 29%, mp 166–168 oC; IR (νmax, cm-1): 3252 and 3186 (NH), 1651 (C=O), 1273 (C=S). 1H NMR (400 MHz, DMSO-d6) δ (ppm): 6.79 (1H, s, OH), 7.22–7.75 (10H, m, Ar-H), 7.88 (1H, s, Ar-H4), 8.26 (2H, s, Ar-H2, Ar-H6), 9.64 (1H, s, N2-H), 10.27 (1H, s, N1-H), 10.55 (1H, N4-H). Anal. Calcd for C23H17F6N3O2S (513.45): C, 53.80; H, 3.34; N, 8.18; S, 6.24%. Found: C, 54.44; H, 3.39; N, 8.29; S, 6.24%. (APCI-, m/z, %): 512 (M+-H)+, 511 (100), 286, 252.

1-(α,α-diphenyl-α-hydroxy)acetyl-4-(2-chloro-5-trifluoromethylphenyl) thiosemicarbazide (2h): White crystals, yield 58%, mp 152–154 ºC; IR (νmax, cm-1): 3302 and 3180 (NH), 1693 (C=O), 1261 (C=S). 1H NMR (400 MHz, DMSO-d6) δ (ppm): 6.65 (1H, s, OH), 7.27–7.86 (13H, m, Ar-H), 10.75 (1H, N4-H), 9.06 (1H, s, N2-H), 10.09 (1H, s, N1-H). Anal. Calcd for C22H17ClF3N3O2S (479.90): C, 55.06; H, 3.57; N, 8.76; S, 6.68%. Found: C, 55.21; H, 3.58; N, 8.72; S, 6.82%.

1-(α,α-diphenyl-α-hydroxy)acetyl-4-(2-trifluoromethylphenyl) thiosemicarbazide (2i): White crystals, yield 64%, mp 168–170 ºC; IR (νmax, cm-1): 3271 and 3194 (NH), 1651 (C=O), 1278 (C=S). 1H NMR (400 MHz, DMSO-d6) δ (ppm): 6.64 (1H, s, OH), 7.14–7.82 (14H, m, Ar-H), 9.02 (1H, N4-H), 9.94 (1H, s, N2-H), 10.55 (1H, s, N1-H). Anal. Calcd for C22H18F3N3O2S (445.45): C, 59.32; H, 4.07; N, 9.43; S, 7.20%. Found: C, 58.36; H, 4.51; N, 8.73; S, 6.80%. (API-ES+, m/z, %): 446 [M+] (100), 428, 368, 302, 296, 279, 257, 225, 192.

1-(α,α-diphenyl-α-hydroxy)acetyl-4-(2-methylsulphanylphenyl) thiosemicarbazide (2j): White crystals, yield 63%, mp 164–166 ºC; IR (νmax, cm-1): 3325 and 3261 (NH), 1651 (C=O), 1255 (C=S). 1H NMR (400 MHz, DMSO-d6) δ (ppm): 2.34 (3H, s, S-CH3), 6.72 (1H, s, OH), 7.11–7.60 (14H, m, Ar-H), 9.00 (1H, N4-H), 9.82 (1H, s, N2-H), 10.53 (1H, s, N1-H). Anal. Calcd for C22H21N3O2S2 (423.55): C, 62.39; H, 5.00; N, 9.92; S, 15.14%. Found: C, 62.21; H, 4.93; N, 9.89; S, 14.92%.

1-(α,α-diphenyl-α-hydroxy)acetyl-4-(1-adamantyl)thiosemicarbazide (2k): White crystals, yield 83%, mp 160–162 oC; IR (νmax, cm-1): 3313 and 3180 (NH), 1651 (C=O), 1282 (C=S). 1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.06 (3H, t,–CH3 of ethanol), 1,48–2.59 (16H, m, adamantyl-H), 3.40–3.53 (2H, m,–CH2 of ethanol), 4.36 (1H, t, OH of ethanol), 6.12 (1H, N4-H), 6.92 (1H, s, OH), 7.23–7.48 (10H, m, Ar-H), 9.13 (1H, s, N2-H), 10.26 (1H, s, N1-H). 13C NMR (100 MHz, DMSO-d6) δ (ppm): 19.21, 29.64, 36.58, 39.51, 39.72, 39.93, 40.14, 40.35, 40.55, 40.77, 41.54 (adamantyl), 53.68 (solvent), 56.76 (NH-adamantyl carbon), 81.05 (C-OH), 128.10, 128.32 (Ar), 144.21 (C=O), 180.00 (C=S). Anal. Calcd for C25H29N3O2S.C2H5OH (481.65): C, 67.33; H, 7.32; N, 8.72; S, 6.66%. Found: C, 67.14; H, 7.07; N, 8.79; S, 6.51%. (API-CI+, m/z, %): 436 [M+] (100), 418, 226, 225, 243.

General procedure for the synthesis of [5-(substitutedamino)- 1,3,4-thiadiazole-2-yl](diphenyl)methanol (3a-b)
A sample of corresponding 1,4-disubstituted-thiosemicarbazides (0.001 mol) was treated with concentrated sulphuric acid at room temperature with constant stirring for 1-2 h. The reaction mixture was poured into ice water. The product was precipitated, filtered and washed with water to afford 3a-b in quantitative yield.

[5-(ethenylamino)-1,3,4-thiadiazole-2-yl](diphenyl)methanol (3a): White powder, yield 81%, mp 202–203 ºC; IR (νmax, cm- 1): 3257 (NH), 1614 (C=N). 1H NMR (400 MHz, DMSO-d6) δ (ppm): 3.66 (t, 2H, =CH2, J= 6.86 Hz, 13.77 Hz), 4.76 (s, 1H, OH), 6.79 (d, 1H, -CH=, J=7.70 Hz), 6.93–7.42 (m, 10 H, ArH), 7.81 ve 8.18 (2s, 1H, NH). Anal. Calcd for C17H15N3OS (309.38): C, 66.00; H, 4.89; N, 13.58; S, 10.36%. Found: C, 65.12; H, 4.86; N, 13.11; S, 10.13%. (API-ES+, m/z, %): 310 [M+] (100), 256, 208, 192.

[5-(benzoylamino)-1,3,4-thiadiazole-2-yl](diphenyl)methanol (3b): Yellow crystals, yield 87%, mp 206–208 oC; IR (νmax, cm-1): 3250 and 3165 (NH), 1600 (C=N). 1H NMR (400 MHz, DMSO-d6) δ (ppm): 7.04–8.17 (m,16 H, ArH ve OH), 12.44 (s, 1H, -NHCO). Anal. Calcd for C22H17N3O2S (387.45): C, 68.20; H, 4.42; N, 10.85; S, 8.28%. Found: C, 68.20; H, 4.61; N, 10.91; S, 8.90%.

General procedure for the synthesis of 3-[hydroxy(diphenyl) methyl]-4-(nonsusbtituted/substituted)-2,4-dihydro-5H- 1,2,4-triazole-5-thione (4a-c)
Sodium hydroxide (2N, 15-20 mL) was added to corresponding 1,4-disubstituted-thiosemicarbazides and refluxed on water bath for 4h. The reaction mixture was neutralized with hydrochloric acid (10%). The precipitate was filtered, washed with water and crystallized from ethanol.

3-[hydroxy(diphenyl)methyl]-2,4-dihydro-5H-1,2,4-triazole- 5-thione (4a): White crystals, yield 49%, mp 258 ºC; IR (νmax, cm-1): 3107 (NH), 1577, 1487 (C=N, NH), 1260 (C-N), 1180 (C=S), 1H NMR (400 MHz, DMSO-d6) δ (ppm): 6.95 (1H, s, OH), 7.11–7.58 (10H, m, Ar-H), 13.20 (1H, s, NH), 13.40 (1H, s, NH). Anal. Calcd for C15H13N3OS.1/2 H2O (292.35): C, 61.62; H, 4.83; N, 14.37; S, 10.97%. Found: C, 60.97; H, 4.60; N, 13.84; S, 10.64%.

3-[hydroxy(diphenyl)methyl]-4-[4-(trifluoromethoxy) phenyl]-2,4-dihydro-5H-1,2,4-triazole-5-thione (4b): White crystals, yield 84%, mp 235–237 ºC; IR (νmax, cm-1): 3169 (NH), 1510, 1558 (C=N, NH), 1271 (C-N), 1155 (C=S). 1H NMR (400 MHz, DMSO-d6) δ (ppm): 6.91 (2H, d, Ar-H, J= 8.9 Hz), 7.00 (1H, s, OH), 7.15 (2H, d, Ar-H, J= 8.2 Hz), 7.18–7.35 (10H, m, Ar-H), 13.49 (1H, s, NH). Anal. Calcd for C22H16F3N3O2S.H2O (461.46): C, 57.26; H, 3.93; N, 9.11; S, 6.95%. Found: C, 58.04; H, 4.03; N, 9.05; S, 6.34%. (API-ES+, m/z, %): 444 [M+], 443, 426, 425, 388, 314, 261, 79.

4-[3,5-bis(trifluoromethyl)phenyl)]-3-[hydroxy(dipenyl) methyl]-2,4-dihydro-5H-1,2,4-triazole-5-thione (4c): White crystals, yield 78%, mp 215–216 oC; IR (νmax, cm-1): 3365 (NH), 1556, 1492 (C=N, NH),1276 (C-N), 1120 (C=S). 1H NMR (400 MHz, DMSO-d6) δ (ppm): 7.12 (1H, s, OH), 7.18–7.34 (10H, m, Ar-H), 7.52 (2H, s, triazole ArC2-H, ArC6-H), 8.07 (1H, s, triazole ArC4-H), 14.14 (1H, s, NH). Anal. Calcd for C23H15F6N3OS (495.44): C, 55.76; H, 3.05; N, 8.48; S, 6.47%. Found: C, 55.86; H, 2.86; N, 8.55; S, 6.19%.

Cytotoxic Activity
The synthesized compounds were tested for their cytotoxic activities. Cell viability and cytotoxic activity profile of the compounds were analyzed using the Cell Proliferation Kit I (MTT) [Roche, Germany]. MTT [3-(4,5-dimethylthiazole-2-yl)-2,5- diphenyltetrazolium bromide] is cleaved to formazan crystals by the “succinate-tetrazolium reductase” system which belongs to the mitochondrial respiratory chain and is active only in viable cells[37-38]. HEK293 cell line was used for the determination of cytotoxic activity. The MTT metabolic assay was carried out in 96-well flat-bottom cell culture plates seeded with 5x103 cells/well. HEK293 cells in 100mM containing LGlutamine without antibiotic Eagle’s MEM (Minimum Essential Medium) and RPMI 1640MEM with 10% FBS (Fetal Bovine Serum).

The following day, media was aspirated and the compounds were solved in DMSO and diluted with medium before they were added to the cell cultures at the concentrations of 5.0 μg/ mL and 10.0 μg/mL. Cells were incubated for 48 hrs at 37ºC, 5.0% CO2. After the incubation period add 10 μL of the MTT labeling reagent (final concentration 0.5 mg/mL) to each well. Incubate the microplate for 4-12 hrs in a humidified atmos phere (e.g. 37ºC, 5.0% CO2) and add 100 μL of the solubilization solution into each well. Allow the plate to stand overnight in the incubator in a humidified atmosphere (e.g. 37ºC, 5.0% CO2), the formazan crystals solubilized. Absorbance of formazan product was measured with spectrophotometrically at 550 and 690 nm.


A series of 1,4-disubstituted-thiosemicarbazides, 2,5-disubstituted- 1,3,4-thiadiazoles and 1,2,4-triazole-5-thiones were synthesized and screened for their cytotoxic activities against HEK293 cell line. The cytotoxicity screening indicated that among the tested compounds 1-(α,α-diphenyl-α-hydroxy) acetyl-4-cyclohexylmethylthiosemicarbazide (2e) exhibited cytotoxic activity.

This study was financially supported by the Marmara University Scientific Research Committee (Project No: SAG-CYLP- 090909-0286).


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13) Karakuş S, Rollas S. Synthesis and antituberculosis activity of new N-phenyl-N´-[4-(5-alkyl/arylamino-1,3,4- thiadiazole-2-yl)phenyl]thioureas. Farmaco 2002; 57: 577–81.

14) Oruç EE, Rollas S, Kandemirli F, Shvets N, Dimongo AS. 1,3,4-Thiadiazole derivatives. Synthesis, structure elucidation and strucuture-antituberculosis activity relationship investigation. J Med Chem 2004; 47: 6760–67.

15) Solak N, Rollas S. Synthesis and antituberculosis activity of 2-(aryl/alkylamino)-5-(4-aminophenyl)-1,3,4-thiadiazoles and their schiff bases. ARKIVOC 2006; 12: 173–181.

16) Zamani K, Faghihi K, Mehraniani MS. Synthesis of some new 2,5-disubstituted 1,3,4-thiadiazoles containing isomeric pyridyl as potent antimicrobial agents. Pol J Pharmacol 2003; 55: 1111–17.

17) Güzeldemirci NU, Küçükbasmacı Ö. Synthesis and antimicrobial activity evaluation of new 1,2,4-triazoles and 1,3,4-thiadiazoles bering imidazo[2,1-b]thiazole moiety. Eur J Med Chem 2010; 45: 63–8.

18) Amir M, Shikha K. Synthesis and anti-inflammatory, analgesic, ulcerogenic and lipid peroxidation activities of some new 2-[(2,6-d,chloroanilino)phenyl]acetic acid derivatives. Eur J Med Chem 2004; 39: 535–45.

19) Rollas S, Kokyan Ş, Koçyiğit-Kaymakçıoğlu B, Özbaş- Turan S, Akbuğa J. Synthesis and evaluation of cytotoxic activities of some substituted isoxazolone derivatives. Marmara Pharm J 2011; 15: 94-9.

20) Matysiak J. Evaluation of antiproliferative effect in vitro of some 2-amino-5-(2,4-dihydroxyphenyl)-1,3,4-thiadiazole derivatives. Chem Pharm Bull 2006; 54: 988–91.

21) Kuş C, Ayhan-Kılcıgil G, Özbey S, Kaynak FB, Kaya M, Çoban T, Can-Eke B. Synthesis and antioxidant properties of novel N-methyl-1,3,4-thiadiazol-2-amine and 4-methyl- 2H-1,2,4-triazole-3(4H)-thione derivatives of benzimidazole class. Bioorg Med Chem 2008; 16: 4294–303.

22) Khan I, Ali S, Hameed S, Rama NH, Hussain MT, Wadood A, Uddin R, Ul-Hag Z, Khan A, Ali S, Choudhary I. Synthesis, antioxidant activities and urease inhibition of some new 1,2,4-triazole and 1,3,4-thiadiazole derivatives. Eur J Med Chem 2010; 45: 5200–207.

23) Siddiqui N, Ahsan W. Triazole incorporated thiazoles as a new class of anticonvulsants: Design, synthesis and in vivo screening. Eur J Med Chem. 2010; 45: 1536-43.

24) Küçükgüzel ŞG, Küçükgüzel İ, Tatar E, Rollas S, Şahin F, Güllüce M, De Clercq E, Kabasakal L. Synthesis of some novel heterocyclic compounds derived from diflunisal hydrazide as potential anti-infective and anti-inflammatory agents. Eur J Med Chem 2007; 42: 893–901.

25) Dündar Y, Çakır B, Küpeli E, Şahin MF, Noyanalpan N. Synthesis of some new 1-acylthiosemicarbazides and 1,2,4-triazol-5-thiones, and their analgesic and anti-inflammatory activities. Turk J Chem 2007; 31: 301–13.

26) El-Emam AA, Ibrahim TM. Synthesis and anti-inflammatory and analgesic activity of some 3-(1-adamantyl)- 4-substituted-5-mercapto-1,2,4-triazoles. Arzneim-Forsh/ Drug Res 1991; 41: 1260–64.

27) Labanauskas L, Udrenaite E, Gaidelis P, Brukstus A. Synthesis of 5-(2,3- and 4-methoxyphenyl)-4H-1,2,4- triazole-3-thiol derivatives exhibiting anti-inflammatory activity. Farmaco 2004; 59: 255–9.

28) Kumar H, Javed SA, Khan SA, Amir M. 1,3,4-Oxadiazole/ thiadiazole and 1,2,4-triazole derivatives of biphenyl- 4-yloxy acetic acid: Synthesis and preliminary evaluation of biological properties. Eur J Med Chem 2008; 43: 2688–98.

29) Colanceska-Ragenovic K, Dimova V, Kakurinov V, Molnar DG, Buzarovcka A. Synthesis, antibacterial and antifungal activity of 4-substituted-5-aryl-1,2,4-triazoles. Molecules 2001; 6: 815-24.

30) Karakuş S, Çoruh U, Barlas-Durgun B, Vázquez-López EM., Özbaş-Turan S, Akbuğa J, Rollas S. Synthesis and cytotoxic activity of some 1,2,4-triazoline-3-thione and 2,5-disubstituted- 1,3,4-thiadiazole derivatives. Marmara Pharm J 2010; 14: 84-90.

31) Mavrova TA, Wesselinova D, Tsenov YA, Denkova P. Synthesis, cytotoxicity and effects of some 1,2,4-triazole and 1,3,4-thiadiazole derivatives on immunocompetent cells. Eur J Med Chem 2009; 44: 63–9.

32) Turan-Zitoni G, Kaplancıklı ZA, Yıldız MT, Chevalent P, Kaya D. Synthesis and antimicrobial activity of 4-phenyl/ cyclohexyl-5-(1-phenoxyethyl)-3-[N-(2-thiazolyl) acetamido]thio-4H-1,2,4-triazole derivatives. Eur J Med Chem 2005; 40:607-13.

33) Doğan HN, Duran A, Rollas S. Synthesis and preliminary anticancer activity of new 1H-4,5-dihydro-3-(3-hydroxy- 2-naphthyl)-4-substituted-1,2,4-triazole-3-thiones. Indian J Chem Sec B 2005; 44: 2301-7.

34) Hou YP, Sun J, Pang ZH, Lv PC, Li DD, Zhang HJ, Zheng EX, Zhao J, Zhu HL. Synthesis and antitumor activity of 1,2,4-triazoles having 1,4-benzodioxan fragment as novel class of potent methionine aminopeptidase type II. Bioorg Med Chem 2011; 19: 5948-54.

35) Akhtar T, Hameed S, Al-Masoudi NA, Khan KM. Synthesis and anti-HIV activity of new chiral 1,2,4-triazoles and 1,3,4-thiadiazoles. Heteroatom Chem 2007; 18: 316–22.

36) Abu Safieh KA, Al-Titi AMS, Zahra JA, Ayoub MT. Oxidative cyclization of arylidene carboxyhydrazides: Synthesis of substituted hydroxydiphenylmethyl-1,3,4- oxadiazoles. Jordan J Chem 2007; 2: 211–8.

37) Mossman T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods 1983; 65: 55-63.

38) Beekman AC, Barentsen AR, Woerdenbag HJ, Uden WV, Pras N. Stereochemistry-dependent cytotoxicity of some artemisinin derivatives. J Nat Prod 1997; 60: 325-30.

39) Ilhan E, Ergenç N, Uzun M, Kaya D. Synthese von 6-Benzyliden-2-(α,α-diphenyl-α-hydroxyacetyl)- thiazolo[3,2-b]-s-triazol-5-onen als potentiell biologisch wirksame Stoffe. Arch Pharm 1994; 327: 825–6.

40) Rollas S. Synthesis and spectroscopic data of some 1,3,4-thiadiazoles. J Fac Pharm İstanbul 1982; 18: 3–12.

41) Rollas S. Synthesis and spectrometric analysis of some 1,2,4-triazoline-5-thiones. J Fac Pharm İstanbul 1981; 17: 155–63.

42) Rollas S. Bazı l-aroil-4-alkil/ariltiyosemikarbazidler I. Doğa Bilim Dergisi 1983; Tıp Cilt 7: 65–73.

43) Ergenç N, Ilhan E, Ötük G. Synthese und biologische Wirkung einiger 1,4-disubstituierter Thiosemicarbazide und deren 1,2,4-Triazol-5-thion-Derivate. Pharmazie 1992; 47: 59-60.

44) Kalyoncuoğlu N, Rollas S, Sür-Altıner D, Yeğenoğlu Y, Anğ Ö. 1-[p-(Benzoylamino)benzoyl]-4-substitutedthiosemicarbazides. Synthesis and antibacterial and antifungal activities. Pharmazie 1992; 47:796-7.

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