Anticancer And Antibacterial Studies Of New Asymmetric Triazoles Biology Essay

A cardinal intermediate 4,5-diphenyl-4H-1,2,4-triazole-3-thiol ( 1 ) was synthesized from base catalyzed dehydrative cyclization of phenyl thiosemicarbazide. The 1,2,4-triazole-3-thiol ( 1 ) was converted to 3-thio acerb hydrazide ( 3a, B ) via 3-thio ethyl esters ( 2a, B ) intermediates.

Then 3-thiol acid hydrazide ( 3a, B ) were converted to asymmetric bis-1,2,4-triazole ( 5a-l ) via thiosemicarbazide intermediate ( 4a-l ) . The synthesized compounds were characterized by IR and NMR spectral surveies.Synthesized compounds 5a-l were evaluated for in vitro antioxidant activity by DPPH extremist scavenging assay method and rating of in vitro anticancer activity by MTT assay method against three malignant neoplastic disease cell lines, HT29 ( human glandular cancer ) , EAC ( Ehrich Ascites carcinoma ) and MDA-231 ( Human chest malignant neoplastic disease ) . All the synthesized compounds were subjected to in vitro antibacterial activity against Bacillus subtilus ( ATCC 6633 ) , Staphylococcus aureus ( ATCC-25923 ) , Escherichia coli ( ATCC-25922 ) , and Pseudomonas aeruginosa ( ATCC- 27853 ) and their zone of suppression is determined.IntroductionCancer is the 2nd prima cause of decease worldwide, following bosom diseases. World Health Organization has estimated 12 million deceases worldwide due to malignant neoplastic disease in 2030. Although advancement has been steadily made in malignant neoplastic disease research to cut down mortality and better endurance, malignant neoplastic disease still accounts for about 1 in every 4 deceases in the universe ( Jemal A et Al 2009 ) . Triazoles are known to hold a big spectrum of possible anticancer, antimitotic and fungicidal belongingss ( yashwant et al 2010 ) .

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Dimeric parallels of assorted heterocyclic compounds are pulling much attending in the recent yesteryear. Many dimeric compounds designed as bis-DNA intercalators were evaluated as anti-cancer agents. Dimers of more Dimers of more lipotropic compounds have shown potent and wide spectrum activity against human solid tumour cell lines both in civilization and as heterografts in bare mice. Some of the bis-intercalators were found to possess high selective toxicity against human colon carcinoma ( Yong et al. , 2005 ; Denny, 2003 ) .

Asymmetrical Synthesis ( Fucheng Qu et.al 1999 ) , anticancer ( Shivarama Holla et. Al 2001, Spicer et al. , 2000 ; Dabholkar and Ansari, 2008 ) , ( Al-Masoudi et al 2004 ) , ( De-Chang Zhang et Al 2006 ) , ( Krzysztof Sztanke et Al. 2008 ) .

Many of the bis- 1,2,4-triazoles have besides been reported to possess broad spectrum of biological activity ( Holla et al. , 2000 ; Holla et al. , 2002 ; Ghorab et al. , 2000 ; Al-Soud and Al-Masoudi, 2004 ) . The triazole derived functions vorozole, letrozole, and anastrozole are non-steroidal drugs used for the intervention of chest malignant neoplastic disease ( Clemons et al. , 2004 ) .

Based on the above literature, the writers have tried to plan the bis-triazole derived functions by integrating assorted alterations utilizing groups such as permutation of other heterocyclic ring at 5th place through oxyalkyl or alkyl linkages ( Demirbas et al 2004 ) , Doubling the heterocyclic ring every bit good as replacing of Cl by F or by ethoxy, methoxy and by isopropyl groups. On the other manus, the effectivity of commercially available anti-microbials has become undependable, due to the outgrowth of immune micro-organisms like methicillin immune Staphylococcus aureus ( MRSA ) , chloroquineresistant Plasmodium falciparam, multi-drug-resistant Mycobacterium TB, and vancomycin-resistant Enterococcus faecium ( VRE ) ( Rostom et al. , 2009 ) . Hence such type of infections continue to be the impulsive force for the hunt and find of novel, more powerful and selective non-traditional anti-microbial agents with the less likelihood of development of cross-resistance. Based on these studies, attempts are being made in the synthesis and biological activity of symmetric and asymmetric Bi heterocyclic compounds, in peculiar 1,2,4- triazoles. In position of these determination, we propose to synthesise asymmetric bis-1,2,4-triazole and measure for in vitro anticancer and antibacterial activity.Consequences and treatmentChemistrySynthesis of the intermediate and aim compounds was accomplished harmonizing to the stairss depicted in Scheme 1.

A cardinal intermediate 4,5-diphenyl-4H-1,2,4-triazole-3-thiol ( 1 ) was synthesized from base catalyzed dehydrative cyclization of phenyl thiosemicarbazide. The compound 4, 5-diphenyl-4H-1,2,4-triazol-3-ylthio-ethylesters ( 2a, B ) were synthesized from 4,5-diphenyl-4H-1, 2, 4-triazole-3-thiol ( 1 ) by handling it with ethyl chloroacetate/ ethyl chloroformate utilizing dry propanone and K2CO3. Refluxing ( 2a, B ) with surplus of hydrazine hydrate in absolute ethyl alcohol afforded several acid hydrazides ( 3a, B ) in good output. The compound ( 4a-l ) was prepared by handling the acidhydrazides ( 3a, B ) with utilizing six different substituted isothiocyanates viz. phenyl, p-flurophenyl, p-chlorophenyl, p-tolyl, isopropyl and p-ethoxyphenyl isothiocyanate. The mark compounds ( 5a-l ) were synthesized by base catalyzed dehydrative cyclization of ( 4a-l ) . The constructions of all the compounds were elucidated on the footing of elemental analysis, IR, 1H-NMR, 13C-NMR, and mass spectral informations.SCHEME OF SYNTHESISThe IR spectra of compound 1 exhibits the absence of signals at 1660 cm?1 corresponds C=O, therefore corroborating the formation of cyclized compound.

Further, the1H-NMR spectrum shows the complex multiplet for aromatic protons appears between 7.02-7.89ppm. The wide vest at 14.12 ppm is due to NH attached to C=S reveals the farther verification the formation of compound. In the 13C-NMR spectrum of the same compound, the signals at 153.22 and 169.

56 ppm were due to the C3 and C5 of the triazole mediety. The chemical displacements at 130.74, 129.86, 128.74 and 127.15 were assigned to the aromatic C atoms of the phenyl ring.

The mass spectrum shows molecular ion extremum ( M+ ) and basal extremum at 253 m/z.The IR spectrum of compounds 2a exhibits the presence of signals at 1728 cm?1 bespeaking the presence of a C=O, therefore corroborating the formation of ester compound. 1H-NMR spectrum of compound 2a the complex multiplet for aromatic protons appears between 7.33-7.56ppm. The vest at 4.

08ppm integrates for 2H of CH2 group ; where as the quadrate at 4.14ppm integrates for 2H of OCH2 group. The three at 1.20ppm histories for the 3H of CH3 group. In the 13C-NMR spectrum of the same compound, the chemical displacement was observed at 169.50 ppm, this was assigned to the carbonyl C of the ester compound.

The signals at 147.62 and 153.10 ppm were due to the C3 and C5 of the triazole mediety. The chemical displacements at 130.77, 129.91, 128.84 and 127.

50ppm were assigned to the aromatic C atoms of the phenyl ring. The signals at 60.06 and 14.13 ppm were assigned to carbon atom of the -CH2 and -CH3 groups of the diethyl carbamoyl side concatenation. The C atoms of -SCH2 of the side concatenation resonated at 32.50 ppm.The IR spectrum of the 3a exhibits the soaking up set for C=O group ( amide ) at 1660 cm?1 and for NH group at 3320 cm-1 indicates the formation of the compound 3a. The 1H-NMR Spectrum 3a shows the complex multiplet for aromatic protons appears between 7.

32-7.55ppm. The doublet at 4.28-4.31ppm integrates for 2H of NH2 group ; whereas the vest at 9.32ppm integrates for 1H of NH. The vest at 3.89ppm histories for the 2H of SCH2 group.

The absence of signals for alkyl group and presence of extremums for NH, NH2 group confirms formation of compound 3a. The 13C-NMR spectrum of the compound 3a showed a signal at 170.34 ppm due to the -CONH functional group. The signal at 153.18 and 147.64 ppm was attributed to the C3 and C5 of the triazole mediety. The chemical displacements at 130.

94, 129.71, 128.80, and 127.18 were assigned to the aromatic C atoms of the phenyl ring. The C atom of -SCH2 of the side concatenation resonated at 40.92 ppm.

The IR spectrum of the 4a exhibits signals at 1703 cm-1 which bespeaking the presence of a C=O. The 1H-NMR spectrum of the 4a shows the complex multiplet for aromatic protons appears between 7.02-7.

55ppm. A vest at 4.12ppm histories for the 2H of SCH2 group and three vest extremums of thiosemicarbazide appears between 10.21-11.

20ppm indicates the formation compounds. This farther confirmed the formation of compound 4a. The 13C-NMR spectrum of the compound 4a showed a signal at 170.34 ppm due to the -CONH functional group, it besides shows the signal at 181.53 due to C=S.

The signal at 153.18 and 147.64 ppm was attributed to the C3 and C5 of the triazole mediety. The chemical displacements at 137.14, 130.

73, 129.34, 128.67, and 127.53, 126.45 and 124.82 were assigned to the aromatic C atoms of the phenyl ring.

The C atom of -SCH2 of the side concatenation resonated at 41.52 ppm.The IR spectrum of the 5a exhibits the absence of signals at 1703 cm?1 bespeaking the deficiency of a C=O, therefore corroborating the formation of cyclized compound. The 1H-NMR spectrum of 4a exhibits the complex multiplet for aromatic protons appears between 7.22-7.55ppm. The wide vest at 13.84ppm is due to NH attached to C=S.

The absence of NH vest of the thiosemicarbazide and presence of NH attached to C=S extremum, farther confirms the formation of compound 5a. The 13C-NMR spectrum of the compound 5a showed a signal at 168.92 and 161.32 ppm which was attributed to the C3 and C5 of one of the triazole mediety and other triazole mediety shows signals at 153.76 and 148.

73 ppm. The chemical displacements at 130.28, 129.61, 128.54, and 127.82ppm were assigned to the aromatic C atoms of the phenyl ring.

The C atom of -SCH2 of the side concatenation resonated at 27.92 ppm.Similar account for delegating the C holds good for the remainder of the compounds. In decision, 1H-NMR and 13C-NMR spectral informations were consistent with the proposed constructions.

The mass spectra of all the triazole derived functions were analyzed under ESI conditions. Molecular ions were observed in the signifier of M+ . Most of the compounds yield abundant molecular ions in the signifier of M+ extremums. Similarly, the elemental analyses of all the compounds have been performed and the informations given under the physical informations in Table 1.Table 1 Physical information of synthesized compoundscompoundRoentgenNMol. wtMol. expressionoutputMP ( 0C )Elemental analysis calculated ( found )CHydrogen1

253C14H11N3S75202-20466.38( 66.

21 )4.38( 4.29 )2a

1339C18H17N3O2S64116-11863.

70( 63.62 )5.05( 4.98 )2b

0312C16H14N3O2S65104-10661.52( 61.63 )4.52( 4.

40 )3a

1325C16H15N5OS72184-18659.06( 58.95 )4.65( 4.55 )3b

0311C15H13N5OS65202-20457.

86( 57.72 )4.21( 4.15 )4aPhenyl1460C23H20N6S2O83212-21459.

98( 59.83 )4.38( 4.23 )4b4-fluorophenyl1478C23H19N6S2OF85240-24257.72( 57.55 )4.00( 3.87 )4c4-chlorophenyl1494C23H19N6S2OCl78232-23455.

81( 55.68 )3.87( 3.69 )4d4-methoxyphenyl1490C24H22N6S2O282252-25458.76( 58.65 )4.52( 4.

41 )4e4-ethoxyphenyl1504C25H24N6S2O275262-26457.91( 57.85 )4.32( 4.21 )4fisopropyl1426C20H22N6S2O72236-23859.38( 59.25 )4.

76( 4.78 )4gPhenyl0446C22H18N6S2O78222-22459.19( 59.30 )3.98( 3.

92 )4h4-fluorophenyl0464C22H17N6S2OF73230-23255.09( 55.24 )4.78( 4.65 )4i4-chlorophenyl0481C22H17N6S2OCl78238-24056.73( 56.87 )3.

94( 4.03 )4j4-methoxyphenyl0476C23H20N6S2O270242-24457.47( 57.65 )4.76( 4.

65 )4k4-ethoxyphenyl0490C24H22N6S2O268254-25655.09( 55.24 )4.56( 4.68 )4lisopropyl0412C19H20N6S2O70218-22058.74( 58.83 )4.92( 4.

78 )5aPhenyl1442C23H18N6S278190-19462.42( 62.30 )4.10( 4.02 )5b4-fluorophenyl1460C23H17FN6S271220-22259.98( 59.

80 )3.72( 3.60 )5c4-chlorophenyl1477C23H17ClN6S274226-22860.

73( 60.58 )4.18( 4.04 )5d4-methoxyphenyl1472C24H20N6OS264232-23461.

00( 60.89 )4.27( 4.15 )5e4-ethoxyphenyl1486C25H22N6OS270242-24461.71( 61.55 )4.56( 4.40 )5fisopropyl1408C20H20N6S267216-21858.

80( 58.45 )4.93( 4.

8205gPhenyl0428C22H16N6S272252-25461.66( 61.47 )3.76( 3.

58 )5h4-fluorophenyl0446C22H15FN6S274210-21259.18( 59.04 )3.39( 3.25 )5i4-chlorophenyl0463C22H15ClN6S270228-23060.43( 60.29 )3.

84( 3.70 )5j4-methoxyphenyl0458C23H18N6OS268236-23860.24( 60.12 )3.96( 3.

78 )5k4-ethoxyphenyl0472C24H20N6OS270226-22861.23( 61.11 )4.

27( 4.15 )5lisopropyl0394C19H18N6S268218-22057.84( 57.69 )4.60( 4.43 )BIOLOGICAL ACTIVITYIn vitro antioxidant activityIn the present survey, asymmetric bis-1,2,4-triazoles derived functions were evaluated for their free extremist scavenging activity utilizing the DPPH extremist check method.

Decrease of DPPH groups can be measured at 517nm ( Roopan et al 2008, Yukesk et al 2006 ) . Different derived functions of bis-1,2,4-triazoles reduced DPPH groups significantly. The activity of bis-1,2,4-triazoles derived functions was compared with ascorbic acid as criterion. Compounds 5a, 5b, 5c, 5d, 5g, 5h, 5i and 5k shows % scavenging activity runing from 53-85 % . The tried compounds shown more than 50 % of extremist scavenging activity were selected for in vitro anticancer activity.Table 2 Antioxidant activity informations of the synthesized compounds ( 5a-l )S.No.

Compounds% Free Radical Scavenging Activityat 40 µg/ml15a8525b7235c6545d5355e3465f4275g6385h6095i64105j28115k69125l3913Ascorbic acid ( venereal disease )94In vitro anticancer activityThe synthesized compounds 5a, 5b, 5c, 5d, 5g, 5h, 5i and 5k were screened for in vitro anticancer activity concentration against three human malignant neoplastic disease cell lines, severally HT29 ( human glandular cancer ) , EAC ( Ehrich Ascites carcinoma ) and MDA-231 ( Human chest malignant neoplastic disease ) by MTT assay method ( Molinari et al. , 2009, Manjula et al. , 2009 ) . 5-flurouracil was used as standard and DMSO was used as solvent control. The per centum suppression and IC50 of all the tested compounds are given in table 3. Based on the IC50 value it has been observed that compounds shows good to hapless anticancer activity at different concentration. Among the tested compound 5a, 5b and 5c shows good activity against EAC malignant neoplastic disease cell line, where as compound 5a and 5b shows moderate activity against HT-29 and MDA-231 malignant neoplastic disease cell line.

The compound 5a with phenyl group at the 4th place of 1,2,4-triazole ring system shows IC50 value 203.36 µg/ml against the EAC ( ehrich ascities carcinoma ) , which is lesser than the standard drug ( IC50 454.48 µg/ml ) . At the 4th place of 1,2,4-triazole ring with 4-flurophenyl permutation 5b shows IC50 value 181.31 µg/ml, with p-methoxy phenyl 5c shows IC50 value 228.32 µg/ml. It is observed that permutation on the phenyl ring increases the anticancer activity against EAC malignant neoplastic disease cell line, where as the compound 5a with phenyl group at the 4th place of 1,2,4-triazole ring system shows IC50 value 55.49 µg/ml against the HT-29 ( glandular cancer ) and IC50 value 98.

57 µg/ml against the MDA-231 ( breast malignant neoplastic disease ) which are more than the standard drug ( IC50 value 10.04 µg/ml ) . At the 4th place of 1,2,4-triazole ring, with 4-flurophenyl permutation 5b shows IC50 value 46.66 µg/ml and 126 µg/ml against HT-29 and MDA-231 malignant neoplastic disease cell lines. It is observed that permutation on the phenyl ring shows moderate activity against HT-29 and MDA-231 malignant neoplastic disease cell line.Table 3: Anticancer activity informations of synthesized compoundsS.No.

Cpd codificationConc.µg/mlHT-29( glandular cancer )EAC( ehrich ascites carcinoma )MDA-231( chest malignant neoplastic disease )% cytoto-xicityIC50µg/ml% cytoto-xicityIC50µg/ml% cytoto-xicity1Standard ( 5-flurouracil )20090.0010.0424.27454.

4880.1710080.9816.3069.355067.8311.4143.262552.

639.8030.131039.643.3819.

8825a20081.2955.4946.54203.

3670.3510073.6434.1760.745064.2927.

6443.122532.1714.1328.731028.

048.7419.1635b20082.7446.6650.39181.3160.3410071.

1338.2753.425062.7429.1340.122542.3917.1227.

311029.348.3417.6345c20054.23166.5642.13228.

3237.8310040.1732.7430.

795029.3424.7318.392517.1315.

149.591010.327.134,5455d20034.73277.7021.

72460.2315.0410028.6316.7815.245024.389.569.

732511.674.325.41102.372.

364.9365g20027.04281.3219.

12604.6236.7310017.7110.2124.

12509.238.6717.13259.845.5611.

13108.323.89-0.3875h20021.04324.7917.12588.7523.

7310017.7112.6520.

12508.2310.6713.13257.848.5610.13106.

325.893.3885i20016.04389.4314.12679.3920.

7310012.7110.6517.12506.237.

6710.13253.843.567.1310-0.321.894.

3895k20012.04465.7612.73712.6516.261008.219.2911.

18503.346.948.174251.782.545.

2910-1.321.322.89In vitro antibacterial activityThe synthesized compounds were evaluated for their in vitro antibacterial activity against S.aureas, B.

substilus, E.coli, P.aeruginosa by cup home base method ( Rostom et al 2009. , Hugo et al 1997 ) . Ampicillin was used as standard and DMSO was used as solvent control.

The antibacterial activity informations of the synthesized compounds are given in Table 4. The tried compounds show good to chair antibacterial activity at 100 ?g/ml concentration with the zone of suppression in the scope of 05-25 millimeter, the standard drug Principen has a zone of suppression in the scope of 28-32 millimeter for the four bacterial species. Among the tried compounds 5b, 5d show good antibacterial activity with zone of suppression in the scope of 21-26 millimeter at 100mg/ml concentration against the tried beings.

5a, 5c, 5h and 5i show moderate antibacterial activity with zone of suppression in the scope of 16-23 millimeter at 100mg/ml concentration against the tried beings. Rest of the derived functions shows weak antibacterial activity with the zone of suppression in the scope of 05-14 millimeter at 100mg/ml concentration against the tried beings.Table 4: Antibacterial activity informations of the synthesized compounds ( 5a-l )S. No.Cpd.

CodeZone of suppression ( in millimeter )S.aureasB.substilusE.coli15a16181725b25232235c20211745d22211955e12131665f08100975g13111485h21191995i201719105j050609115k121610125l17191513Ampicillin282726DecisionA series 5- [ ( 4,5-diphenyl-4H-1,2,4-triazol-3-ylthio ) methyl ] -4-substituted-4H-1,2,4-triazole-3-thiol ( 5a-f ) , 5- ( 4,5-diphenyl-4H-1,2,4-triazol-3-ylthio ) -4-substituted-4H-1,2,4-triazole-3-thiol ( 5g-l ) were synthesized as per the strategy I.

The synthesized compounds were characterized by IR, 1H-NMR informations and mass spectral informations and consistent with the proposed construction. The in vitro anticancer activity of the synthesized compounds was determined by MTT assay method against the three human malignant neoplastic disease cell line viz. HT-29 ( human glandular cancer ) , EAC ( ehrich ascites carcinoma ) and MDA-231 ( human chest malignant neoplastic disease ) and the synthesized compounds were evaluated for their in vitro antibacterial activity against S.aureas, B.substilus, E.

coli, P.aeruginosa by cup home base method. The antimicrobic and cytotoxic consequences suggest that synthesized compounds 5a and 5b could be considered as possible double antimicrobial-anticancer campaigners that deserve farther probe and derivatization in order to research the range and restriction of their biological activities.ExperimentalMaterials and methodsThe thaw points were determined in unfastened glass capillaries and are uncorrected. IR spectra were recorded on Shimadzu FT-IR 8400-S spectrophotometer by KBr pellet technique. Elemental analyses were performed and found values are within 0.4 % of theoretical values unless otherwise noted.

1H-NMR and 13C-NMR spectra were recorded on AMX-400 NMR spectrophotometer at 400 MHz utilizing DMSO-d6 as the dissolver and tetramethylsilane ( TMS ) as internal criterion. The chemical displacements are expressed in ? ppm. The splitting forms were designated as follows ; s: vest ; vitamin D: doublet ; Q: four ; m: multiplet. LCMS were recorded by utilizing Shimadzu LCMS-2010A instrument by ESI.Synthesis of 4, 5-diphenyl-4H-1,2,4-triazole-3-thiol ( 1 )Phenyl thiosemicarbazide ( 0.

01 mole, 2.71 g ) was added part wise to 15 milliliter of 2N NaOH solution and ensuing solution was refluxed for 6 h. After the completion of reaction, the reaction mixture was allowed to chill and filtered. The filtrate was acidified with 2N HCl. The solid obtained by acidification was filtered, washed with H2O, dried and recrystallized from absolute ethyl alcohol. The physical information is presented in Table 1.IR ( cm-1 ) : 3103 ( NH ) , 3031 ( Ar CH ) and 1546 ( C=N ) ; 1H-NMR ? : 14.12 ( s, 1H, SH ) , 7.

02-7.89 ( m, 10H, Ar-CH ) ; 13C-NMR ? : 153.22 ( C3 of triazole ) , 169. 56 ( C5 of triazole ) , 130.74 ( C1 of phenyl ring ) , 129.

86 ( C2 and C6 of phenyl ring ) , 128.74 ( C3 and C5 of phenyl ring ) , 127.15 ( C4 of phenyl ring ) . MS m/z: 253.Synthesis of Ethyl 2- ( 4, 5-diphenyl-4H-1,2,4-triazol-3-ylthio ) ethanoate ( 2a )A mixture of 1 ( 0.01 mole, 2.53 g ) , ethyl chloroacetate ( 0.01 mole,1.

22 milliliter ) and 1 g of anhydrous K carbonate in 50 milliliters dry propanone were refluxed for 8 H on H2O bath. The reaction mixture was cooled and the dissolver removed under decreased force per unit area. The residuary mass was triturated with ice H2O to take K carbonate and extracted with quintessence ( 3 – 50 milliliter ) and the quintessence bed was washed with 10 % Na hydroxide solution ( 3 – 30 milliliter ) followed by H2O ( 3 – 30 milliliter ) and so dried over anhydrous Na sulphate and evaporated to dryness to acquire rough solid. The solid obtained was filtered, dried and recrystallized from absolute ethyl alcohol. The physical information is presented in Table 1.IR ( cm-1 ) : 1728 ( C=O ) and 3031 ( Ar CH ) ; 1H-NMR ? : 7.33-7.56 ( m, 10H, Ar-H ) , 4.

08 ( s, 2H, SCH2 ) , 4.14 ( Q, 2H, CH2 ) , 1.20 ( T, 3H, CH3 ) ; 13C-NMR ? : 169.50 ( C=O ) , 147.62 ( C3 of triazole ) , 153.10 ( C5 of triazole ) , 130.77 ( C1 of phenyl ring ) , 129.91 ( C2 and C6 of phenyl ring ) , 128.

84 ( C3 and C5 of phenyl ring ) , 127.50 ( C4 of phenyl ring ) , 60.06 ( -CH2 ) , 14.

13 ( -CH3 ) , 32.50 ( -SCH2 ) ; MS m/z: 339.Synthesis of O-ethyl S-4,5-diphenyl-4H-1,2,4-triazol-3-yl carbonothioate ( 2b )The experimental process was repeated as with 2b utilizing 1 ( 0.01 mole, 2.

53 g ) , ethyl chloroformate ( 0.01 mole, 1.22 milliliter ) and anhydrous K2CO3 in 50 milliliters dry propanone. The physical information is presented in Table 1.

IR ( cm-1 ) : 1763 ( C=O ) and 3064 ( Ar CH ) ; 1H-NMR ? : 7.39-7.63 ( m, 10H, Ar-H ) , 4.32 ( Q, 2H, CH2 ) , 1.32 ( T, 3H, CH3 ) ; 13C-NMR ? : 168.54 ( C=O ) , 147.

34 ( C3 of triazole ) , 154.56 ( C5 of triazole ) , 130.89 ( C1 of phenyl ring ) , 129.78 ( C2 and C6 of phenyl ring ) , 128.23 ( C3 and C5 of phenyl ring ) , 127.85 ( C4 of phenyl ring ) , 61.

85 ( -CH2 ) , 14.89 ( -CH3 ) ; MS m/z: 312.Synthesis of 2- ( 4,5-diphenyl-4H-1,2,4-triazol-3-ylthio ) acetohydrazide ( 3a )A mixture of 2a ( 0.1 mole, 3.39 g ) and hydrazine hydrate 99 % ( 0.1 mole, 5 milliliter ) in 25 milliliter of ethyl alcohol was reflux for 6 H on H2O bath. The surplus of dissolver was removed and reaction mixture was left nightlong at room temperature and the solid separated was collected by filtration. The solid obtained was filtered, dried and recrystallized from absolute ethyl alcohol.

The physical information is presented in Table 1.IR ( cm-1 ) : 1660 ( C=O, amide ) , 3320 ( NH ) ; 1H-NMR ? : 9.32 ( s, 1H, NH ) , 7.32-7.55 ( m, 10H, Ar-H ) , 4.28-4.

31 ( vitamin D, 2H, NH2 ) , 3.89 ( s, 2H, SCH2 ) ; 13C-NMR ? : 170.34 ( -CONH ) , 153.18 ( C3 of triazole ) , 147.64 ( C5 of triazole ) , 130.94 ( C1 of phenyl ring ) , 129.71 ( C2 and C6 of phenyl ring ) , 128.80 ( C3 and C5 of phenyl ring ) , 127.18 ( C4 of phenyl ring ) , 40.92 ( -SCH2 ) ; MS m/z: 325.Synthesis of 2- ( 4,5-diphenyl-4H-1,2,4-triazol-3-ylthio ) hydrazide ( 3b )The experimental process was repeated as with 3b utilizing 2b ( 0.1 mole, 3.39 g ) and hydrazine hydrate ( 0.1mole, 5 milliliter, ) in absolute ethyl alcohol ( 25 milliliter ) . The physical information is presented in Table 1.IR ( cm-1 ) : 1697 ( C=O, amide ) , 3101 ( NH ) ; 1H-NMR ? : 10.02. ( s, 1H, NH ) , 7.34-7.82 ( m, 10H, Ar-H ) , 4.16-4.45 ( vitamin D, 2H, NH2 ) ; 13C-NMR ? : 169.72 ( -CONH ) , 153.178 ( C3 of triazole ) , 147.28 ( C5 of triazole ) , 131.64 ( C1 of phenyl ring ) , 129.59 ( C2 and C6 of phenyl ring ) , 128.67 ( C3 and C5 of phenyl ring ) , 127.89 ( C4 of phenyl ring ) , 40.26 ( -SCH2 ) ; MS m/z: 311.Synthesis of the 1- [ 2- ( 4,5-diphenyl-4H-1,2,4-triazole-3-ylthio ) ethanoyl group ] -4-phenyl thiosemicarbazide ( 4a ) .An equimolar measure of 3a ( 0.01 mole, 3.25 g ) and phenyl isothiocyanate were individually dissolved in minimal measure of absolute ethyl alcohol. The solution of phenyl isothiocyanate was poured into the solution of hydrazide with uninterrupted stirring. The reaction mixture was refluxed for 8 h. The white solid obtained on chilling the reaction mixture to room temperature was filtered, dried and recrystallized from methylene chloride. The physical information is presented in Table 1.IR ( cm-1 ) : 1243 ( C=S ) , 1703 ( C=O ) ; 1H-NMR ? : 11.20 ( vitamin D, 1H, NH ) , 10.80 ( vitamin D, 1H, NH ) , 10.21 ( s, 1H, NH ) , 7.02-7.55 ( m, 15H, Ar-H ) , 4.12 ( s, 2H, SCH2 ) ; 13C-NMR ? : 170.34 ( -CONH ) , 181.53 ( C=S ) , 153.18 ( C3 of triazole ) , 147.64 ( C5 of triazole ) , 137.14 ( C1 of phenyl ring ) , 130.73 ( C2 and C6 of phenyl ring ) , 129.34 ( C3 and C5 of phenyl ring ) , 128.67 ( C4 of phenyl ring ) , 41.52 ( -SCH2 ) ; MS m/z: 460.Synthesis of 1- [ 2- ( 4,5-diphenyl-4H-1,2,4-triazole-3-ylthio ) ethanoyl group ] -4- ( 4-flurophenyl ) thiosemicarbazide ( 4b )The experimental process was repeated as with 3a ( 0.01 mole, 3.25 g ) and 4-flurophenyl isothiocyanate in 25 milliliters absolute ethyl alcohol. The physical information is presented in Table 1.IR ( cm-1 ) : 1256 ( C=S ) , 1675 ( C=O ) ; 1H-NMR ? : 11.12 ( vitamin D, 1H, NH ) , 10.65 ( vitamin D, 1H, NH ) , 10.13 ( s, 1H, NH ) , 7.10-7.63 ( m, 14H, Ar-H ) , 4.21 ( s, 2H, SCH2 ) ; 13C-NMR ? : 170.68 ( -CONH ) , 180.63 ( C=S ) , 153.91 ( C3 of triazole ) , 148.23 ( C5 of triazole ) , 137.23 ( C1 of phenyl ring ) , 130.78 ( C2 and C6 of phenyl ring ) , 128.69 ( C3 and C5 of phenyl ring ) , 128.52 ( C4 of phenyl ring ) , 41.62 ( -SCH2 ) ; MS m/z: 478.Synthesis of 1- [ 2- ( 4,5-diphenyl-4H-1,2,4-triazole-3-ylthio ) ethanoyl group ] -4- ( 4-chlorophenyl ) thiosemicarbazide ( 4c )The experimental process was repeated as with 3a ( 0.01 mole, 3.25 g ) and 4-chlorophenyl isothiocyanate in 25 milliliters absolute ethyl alcohol. The physical information is presented in Table 1.IR ( cm-1 ) : 1248 ( C=S ) , 1692 ( C=O ) ; 1H-NMR ? : 11.63 ( vitamin D, 1H, NH ) , 10.73 ( vitamin D, 1H, NH ) , 10.76 ( s, 1H, NH ) , 7.22-7.86 ( m, 14H, Ar-H ) , 4.14 ( s, 2H, SCH2 ) ; 13C-NMR ? : 170.67 ( -CONH ) , 180.29 ( C=S ) , 152.45 ( C3 of triazole ) , 148.97 ( C5 of triazole ) , 137.38 ( C1 of phenyl ring ) , 131.25 ( C2 and C6 of phenyl ring ) , 128.17 ( C3 and C5 of phenyl ring ) , 127.64 ( C4 of phenyl ring ) , 40.18 ( -SCH2 ) ; MS m/z: 494.Synthesis of 1- [ 2- ( 4,5-diphenyl-4H-1,2,4-triazole-3-ylthio ) ethanoyl group ] -4- ( 4-methoxyphenyl ) thiosemicarbazide ( 4d )The experimental process was repeated as with 3a ( 0.01 mole, 3.25 g ) and 4-methoxyphenyl isothiocyanate in 25 milliliters absolute ethyl alcohol. The physical information is presented in Table 1.IR ( cm-1 ) : 1246 ( C=S ) , 1682 ( C=O ) ; 1H-NMR ? : 11.15 ( vitamin D, 1H, NH ) , 10.45 ( vitamin D, 1H, NH ) , 10.72 ( s, 1H, NH ) , 7.19-7.76 ( m, 14H, Ar-H ) , 4.10 ( s, 2H, SCH2 ) , 3.73 ( s, 3H, OCH3 ) ; 13C-NMR ? : 169.57 ( -CONH ) , 181.32 ( C=S ) , 152.87 ( C3 of triazole ) , 147.67 ( C5 of triazole ) , 137.92 ( C1 of phenyl ring ) , 130.75 ( C2 and C6 of phenyl ring ) , 128.78 ( C3 and C5 of phenyl ring ) , 127.17 ( C4 of phenyl ring ) , 40.78 ( -SCH2 ) , 55.93 ( CH3 ) ; MS m/z: 490.Synthesis of 1- [ 2- ( 4,5-diphenyl-4H-1,2,4-triazole-3-ylthio ) ethanoyl group ] -4- ( 4-ethoxyphenyl ) thiosemicarbazide ( 4e )The experimental process was repeated as with 3a ( 0.01 mole, 3.25 g ) and 4-ethoxyphenyl isothiocyanate in 25 milliliters absolute ethyl alcohol. The physical information is presented in Table 1.IR ( cm-1 ) : 1253 ( C=S ) , 1690 ( C=O ) ; 1H-NMR ? : 11.43 ( vitamin D, 1H, NH ) , 10.78 ( vitamin D, 1H, NH ) , 10.67 ( s, 1H, NH ) , 7.25-7.76 ( m, 14H, Ar-H ) , 4.24 ( s, 2H, SCH2 ) , 3.98 ( Q, 2H, CH2 ) , 1.33 ( T, 3H, CH3 ) ; 13C-NMR ? : 170.27 ( -CONH ) , 180.54 ( C=S ) , 152.12 ( C3 of triazole ) , 148.74 ( C5 of triazole ) , 137.14 ( C1 of phenyl ring ) , 129.29 ( C2 and C6 of phenyl ring ) , 128.61 ( C3 and C5 of phenyl ring ) , 127.56 ( C4 of phenyl ring ) , 41.36 ( -SCH2 ) , 64.72 ( OCH2 ) , 14.8 ( CH3 ) ; MS m/z: 504.Synthesis of 1- [ 2- ( 4,5-diphenyl-4H-1,2,4-triazole-3-ylthio ) ethanoyl group ] -4- ( isopropyl ) thiosemicarbazide ( 4f )The experimental process was repeated as with 3a ( 0.01 mole, 3.25 g ) and isopropyl isothiocyanate in 25 milliliters absolute ethyl alcohol. The physical information is presented in Table 1.IR ( cm-1 ) : 1250 ( C=S ) , 1685 ( C=O ) ; 1H-NMR ? : 11.37 ( vitamin D, 1H, NH ) , 10.21 ( vitamin D, 1H, NH ) , 10.65 ( s, 1H, NH ) , 7.12-7.63 ( m, 10H, Ar-H ) , 4.18 ( s, 2H, SCH2 ) , 3.97 ( m, 1H, CH ) , 1.36 ( vitamin D, 6H, 2CH3 ) ; 13C-NMR ? : 169.94 ( -CONH ) , 181.23 ( C=S ) , 151.34 ( C3 of triazole ) , 148.67 ( C5 of triazole ) , 137.84 ( C1 of phenyl ring ) , 129.78 ( C2 and C6 of phenyl ring ) , 127.45 ( C3 and C5 of phenyl ring ) , 126.32 ( C4 of phenyl ring ) , 40.65 ( -SCH2 ) , 48.23 ( CH ) , 23.54 ( CH3 ) ; MS m/z: 426.Synthesis of 1- [ 2- ( 4,5-diphenyl-4H-1,2,4-triazole-3-ylthio ) formyl ] -4-phenyl thiosemicarbazide ( 4g )The experimental process was repeated as with 3b ( 0.01 mole, 3.25 g ) and phenyl isothiocyanate in 25 milliliters absolute ethyl alcohol. The physical information is presented in Table 1.IR ( cm-1 ) : 1247 ( C=S ) , 1688 ( C=O ) ; 1H-NMR ? : 11.02 ( vitamin D, 1H, NH ) , 10.45 ( vitamin D, 1H, NH ) , 10.86 ( s, 1H, NH ) , 7.24-7.74 ( m, 15H, Ar-H ) ; 13C-NMR ? : 169.78 ( -CONH ) , 180.46 ( C=S ) , 151.78 ( C3 of triazole ) , 148.23 ( C5 of triazole ) , 137.62 ( C1 of phenyl ring ) , 128.28 ( C2 and C6 of phenyl ring ) , 127.71 ( C3 and C5 of phenyl ring ) , 126.34 ( C4 of phenyl ring ) ; MS m/z: 446.Synthesis of 1- [ 2- ( 4,5-diphenyl-4H-1,2,4-triazole-3-ylthio ) formyl ] -4- ( 4-fluorophenyl ) thiosemicarbazide ( 4h )The experimental process was repeated as with 3b ( 0.01 mole, 3.25 g ) and 4-fluorophenyl isothiocyanate in 25 milliliters absolute ethyl alcohol. The physical information is presented in Table 1.IR ( cm-1 ) : 1245 ( C=S ) , 1683 ( C=O ) ; 1H-NMR ? : 11.16 ( vitamin D, 1H, NH ) , 10.54 ( vitamin D, 1H, NH ) , 10.87 ( s, 1H, NH ) , 7.12-7.64 ( m, 14H, Ar-H ) ; 13C-NMR ? : 168.58 ( -CONH ) , 181.49 ( C=S ) , 150.23 ( C3 of triazole ) , 148.67 ( C5 of triazole ) , 137.78 ( C1 of phenyl ring ) , 129.18 ( C2 and C6 of phenyl ring ) , 128.12 ( C3 and C5 of phenyl ring ) , 127.82 ( C4 of phenyl ring ) ; MS m/z: 464.Synthesis of 1- [ 2- ( 4,5-diphenyl-4H-1,2,4-triazole-3-ylthio ) formyl ] -4- ( 4-chlorophenyl ) thiosemicarbazide ( 4i )The experimental process was repeated as with 3b ( 0.01 mole, 3.25 g ) and 4-chlorophenyl isothiocyanate in 25 milliliters absolute ethyl alcohol. The physical information is presented in Table 1.IR ( cm-1 ) : 1254 ( C=S ) , 1702 ( C=O ) ; 1H-NMR ? : 11.23 ( vitamin D, 1H, NH ) , 10.35 ( vitamin D, 1H, NH ) , 10.97 ( s, 1H, NH ) , 7.32-7.81 ( m, 14H, Ar-H ) ; 13C-NMR ? : 168.17 ( -CONH ) , 180.15 ( C=S ) , 151.75 ( C3 of triazole ) , 148.67 ( C5 of triazole ) , 137.43 ( C1 of phenyl ring ) , 130.14 ( C2 and C6 of phenyl ring ) , 129.23 ( C3 and C5 of phenyl ring ) , 128.65 ( C4 of phenyl ring ) ; MS m/z: 481.Synthesis of 1- [ 2- ( 4,5-diphenyl-4H-1,2,4-triazole-3-ylthio ) formyl ] -4- ( 4-methoxyphenyl ) thiosemicarbazide ( 4j )The experimental process was repeated as with 3b ( 0.01 mole, 3.25 g ) and 4-methoxyphenyl isothiocyanate in 25 milliliters absolute ethyl alcohol. The physical information is presented in Table 1.IR ( cm-1 ) : 1252 ( C=S ) , 1689 ( C=O ) ; 1H-NMR ? : 11.15 ( vitamin D, 1H, NH ) , 10.43 ( vitamin D, 1H, NH ) , 10.83 ( s, 1H, NH ) , 7.23-7.78 ( m, 14H, Ar-H ) , 3.79 ( s, 3H, OCH3 ) ; 13C-NMR ? : 169.23 ( -CONH ) , 181.34 ( C=S ) , 152.45 ( C3 of triazole ) , 148.74 ( C5 of triazole ) , 137.12 ( C1 of phenyl ring ) , 131.56 ( C2 and C6 of phenyl ring ) , 129.92 ( C3 and C5 of phenyl ring ) , 128.34 ( C4 of phenyl ring ) , 54.27 ( OCH3 ) ; MS m/z: 476.Synthesis of 1- [ 2- ( 4,5-diphenyl-4H-1,2,4-triazole-3-ylthio ) formyl ] -4- ( 4-ethoxyphenyl ) thiosemicarbazide ( 4k )The experimental process was repeated as with 3b ( 0.01 mole, 3.25 g ) and 4-ethoxyphenyl isothiocyanate in 25 milliliters absolute ethyl alcohol. The physical information is presented in Table 1.IR ( cm-1 ) : 1243 ( C=S ) , 1693 ( C=O ) ; 1H-NMR ? : 11.27 ( vitamin D, 1H, NH ) , 10.56 ( vitamin D, 1H, NH ) , 10.79 ( s, 1H, NH ) , 7.28-7.78 ( m, 14H, Ar-H ) , 3.79 ( Q, 2H, CH2 ) , 1.12 ( T, 3H, CH3 ) ; 13C-NMR ? : 169.83 ( -CONH ) , 180.73 ( C=S ) , 151.76 ( C3 of triazole ) , 148.63 ( C5 of triazole ) , 137.18 ( C1 of phenyl ring ) , 132.38 ( C2 and C6 of phenyl ring ) , 129.72 ( C3 and C5 of phenyl ring ) , 128.67 ( C4 of phenyl ring ) , 65.27 ( OCH2 ) , 15.34 ( CH3 ) ; MS m/z: 490.Synthesis of 1- [ 2- ( 4,5-diphenyl-4H-1,2,4-triazole-3-ylthio ) formyl ] -4-isopropyl thiosemicarbazide ( 4l )The experimental process was repeated as with 3b ( 0.01 mole, 3.25 g ) and isopropyl isothiocyanate in 25 milliliters absolute ethyl alcohol. The physical information is presented in Table 1.IR ( cm-1 ) : 1245 ( C=S ) , 1690 ( C=O ) ; 1H-NMR ? : 11.32 ( vitamin D, 1H, NH ) , 10.23 ( vitamin D, 1H, NH ) , 10.84 ( s, 1H, NH ) , 7.37-7.86 ( m, 10H, Ar-H ) , 4.04 ( m, 1H, CH ) , 1.43 ( vitamin D, 6H, 2CH3 ) ; 13C-NMR ? : 169.74 ( -CONH ) , 181.45 ( C=S ) , 150.67 ( C3 of triazole ) , 148.57 ( C5 of triazole ) , 137.39 ( C1 of phenyl ring ) , 131.63 ( C2 and C6 of phenyl ring ) , 129.75 ( C3 and C5 of phenyl ring ) , 128.73 ( C4 of phenyl ring ) , 48.56 ( CH ) , 23.73 ( CH3 ) ; MS m/z: 412.Synthesis 5- [ ( 4,5-diphenyl-4H-1,2,4-triazol-3-ylthio ) methyl ] -4-phenyl-4H- 1,2,4- triazole-3-thiol ( 5a ) .The compound 4a ( 0.01 mole, 4.60 g ) was added part wise to 15 milliliter of 2N NaOH solution and ensuing solution was refluxed for 6 h. After the completion of reaction, the mixture was allowed to chill and filtered. The filtrate was acidified with 2N HCl. The solid obtained by acidification was filtered, washed with H2O, dried and recrystallized from absolute ethyl alcohol. The physical information is presented in Table 1.IR ( cm-1 ) : 1496 ( C=N ) ; 1H-NMR ? : 13.84 ( s, 1H, SH ) , 7.22-7.55 ( m, 15H, Ar-H ) , 4.27 ( s, 2H, SCH2 ) ; 13C-NMR ? : 168.92 ( C3 of triazole ) , 161.32 ( C5 of triazole ) , 130.28 ( C1 of phenyl ring ) , 129.61 ( C2 and C6 of phenyl ring ) , 128.54 ( C3 and C5 of phenyl ring ) , 127.82 ( C4 of phenyl ring ) , 27.92 ( -SCH2 ) ; MS m/z: 442.Synthesis of 5- [ ( 4,5-diphenyl-4H-1,2,4-triazol-3-ylthio ) methyl ] -4- ( 4-fluorophenyl ) -4H-1,2,4-triazole-3-thiol ( 5b )The experimental process was repeated as with 4b ( 0.01 mole, 4.78 g ) and 2N NaOH and 2N HCl. The physical information is presented in Table 1.IR ( cm-1 ) : 1514 ( C=N ) ; 1H-NMR ? : 13.83 ( s, 1H, SH ) , 7.25-7.55 ( m, 14H, Ar-H ) , 4.16 ( s, 2H, SCH2 ) ; 13C-NMR ? : 167.73 ( C3 of triazole ) , 154.67 ( C5 of triazole ) , 131.43 ( C1 of phenyl ring ) , 130.34 ( C2 and C6 of phenyl ring ) , 129.23 ( C3 and C5 of phenyl ring ) , 128.78 ( C4 of phenyl ring ) , 28.23 ( -SCH2 ) ; MS m/z: 460.Synthesis of 5- [ ( 4,5-diphenyl-4H-1,2,4-triazol-3-ylthio ) methyl ] -4- ( 4-chlorophenyl ) -4H-1,2,4-triazole-3-thiol ( 5c )The experimental process was repeated as with 4c ( 0.01 mole, 4.94 g ) and 2N NaOH and 2N HCl. The physical information is presented in Table 1.IR ( cm-1 ) : 1512 ( C=N ) ; 1H-NMR ? : 13.72 ( s, 1H, SH ) , 7.21-7.54 ( m, 14H, Ar-H ) , 4.25 ( s, 2H, SCH2 ) ; 13C-NMR ? : 167.56 ( C3 of triazole ) , 149.35 ( C5 of triazole ) , 132.24 ( C1 of phenyl ring ) , 130.45 ( C2 and C6 of phenyl ring ) , 129.56 ( C3 and C5 of phenyl ring ) , 128.86 ( C4 of phenyl ring ) , 27.16 ( -SCH2 ) ; MS m/z: 477.Synthesis of 5- [ ( 4,5-diphenyl-4H-1,2,4-triazol-3-ylthio ) methyl ] -4- ( 4-methoxyphenyl ) -4H-1,2,4-triazole-3-thiol ( 5d )The experimental process was repeated as with 4d ( 0.01 mole, 4.90 g ) and 2N NaOH and 2N HCl. The physical information is presented in Table 1.IR ( cm-1 ) : 1496 ( C=N ) ; 1H-NMR ? : 13.67 ( s, 1H, SH ) , 7.31-7.65 ( m, 14H, Ar-H ) , 4.30 ( s, 2H, SCH2 ) , 3.79 ( s, 3H, OCH3 ) ; 13C-NMR ? : 167.56 ( C3 of triazole ) , 149.32 ( C5 of triazole ) , 131.47 ( C1 of phenyl ring ) , 130.87 ( C2 and C6 of phenyl ring ) , 129.27 ( C3 and C5 of phenyl ring ) , 128.74 ( C4 of phenyl ring ) , 27.91 ( -SCH2 ) , 55.92 ( -0CH3 ) ; MS m/z: 472.Synthesis of 5- [ ( 4,5-diphenyl-4H-1,2,4-triazol-3-ylthio ) methyl ] -4- ( 4-ethoxyphenyl ) -4H-1,2,4-triazole-3-thiol ( 5e )The experimental process was repeated as with 4e ( 0.01 mole, 5.04 g ) and 2N NaOH and 2N HCl. The physical information is presented in Table 1.IR ( cm-1 ) : 1493 ( C=N ) ; 1H-NMR ? : 13.73 ( s, 1H, SH ) , 7.26-7.73 ( m, 14H, Ar-H ) , 4.18 ( s, 2H, SCH2 ) , 3.98 ( Q, 2H, OCH2 ) , 1.33 ( T, 3H, CH3 ) ; 13C-NMR ? : 168.12 ( C3 of triazole ) , 149.83 ( C5 of triazole ) , 132.36 ( C1 of phenyl ring ) , 131.34 ( C2 and C6 of phenyl ring ) , 129.28 ( C3 and C5 of phenyl ring ) , 128.47 ( C4 of phenyl ring ) , 27.23 ( -SCH2 ) , 64.75 ( -CH2 ) , 14.82 ( -CH3 ) ; MS m/z: 486.Synthesis of 5- [ ( 4,5-diphenyl-4H-1,2,4-triazol-3-ylthio ) methyl ] -4-isopropyl-4H-1,2,4-triazole-3-thiol ( 5f )The experimental process was repeated as with 4f ( 0.01 mole, 4.26 g ) and 2N NaOH and 2N HCl. The physical information is presented in Table 1.IR ( cm-1 ) : 1489 ( C=N ) ; 1H-NMR ? : 13.53 ( s, 1H, SH ) , 7.28-7.56 ( m, 10H, Ar-H ) , 4.41 ( s, 2H, SCH2 ) , 4.01 ( m, 1H, CH ) , 1.45 ( vitamin D, 6H, 2CH3 ) ; 13C-NMR ? : 168.67 ( C3 of triazole ) , 149.38 ( C5 of triazole ) , 132.63 ( C1 of phenyl ring ) , 131.43 ( C2 and C6 of phenyl ring ) , 129.82 ( C3 and C5 of phenyl ring ) , 128.42 ( C4 of phenyl ring ) , 27.41 ( -SCH2 ) ; MS m/z: 408.Synthesis of 5- ( 4,5-diphenyl-4H-1,2,4-triazol-3-ylthio ) -4-phenyl-4H-1,2,4-triazole-3-thiol ( 5g )The experimental process was repeated as with 4g ( 0.01 mole, 4.46 g ) and 2N NaOH and 2N HCl. The physical information is presented in Table 1.IR ( cm-1 ) : 1545 ( C=N ) ; 1H-NMR ? : 13.72 ( s, 1H, SH ) , 7.25-7.63 ( m, 15H, Ar-H ) ; 13C-NMR ? : 168.34 ( C3 of triazole ) , 160.23 ( C5 of triazole ) , 131.43 ( C1 of phenyl ring ) , 129.16 ( C2 and C6 of phenyl ring ) , 128.73 ( C3 and C5 of phenyl ring ) , 127.28 ( C4 of phenyl ring ) ; MS m/z: 428.Synthesis of 5- ( 4,5-diphenyl-4H-1,2,4-triazol-3-ylthio ) -4- ( 4-fluorophenyl ) -4H-1,2,4-triazole-3-thiol ( 5h )The experimental process was repeated as with 4h ( 0.01 mole, 4.64 g ) and 2N NaOH and 2N HCl. The physical information is presented in Table 1.IR ( cm-1 ) : 1545 ( C=N ) ; 1H-NMR ? : 13.34 ( s, 1H, SH ) , 7.21-7.56 ( m, 14H, Ar-H ) ; 13C-NMR ? : 167.37 ( C3 of triazole ) , 154.76 ( C5 of triazole ) , 131.34 ( C1 of phenyl ring ) , 130.62 ( C2 and C6 of phenyl ring ) , 129.32 ( C3 and C5 of phenyl ring ) , 128.87 ( C4 of phenyl ring ) ; MS m/z: 446.Synthesis of 5- ( 4,5-diphenyl-4H-1,2,4-triazol-3-ylthio ) -4- ( 4-chlorophenyl ) -4H-1,2,4-triazole-3-thiol ( 5i )The experimental process was repeated as with 4i ( 0.01 mole, 4.81 g ) and 2N NaOH and 2N HCl. The physical information is presented in Table 1.IR ( cm-1 ) : 1497 ( C=N ) ; 1H-NMR ? : 14.08 ( s, 1H, SH ) , 7.17-7.49 ( m, 14H, Ar-H ) ; 13C-NMR ? : 167.56 ( C3 of triazole ) , 149.35 ( C5 of triazole ) , 132.24 ( C1 of phenyl ring ) , 130.45 ( C2 and C6 of phenyl ring ) , 129.56 ( C3 and C5 of phenyl ring ) , 128.86 ( C4 of phenyl ring ) ; MS m/z: 463.Synthesis of 5- ( 4,5-diphenyl-4H-1,2,4-triazol-3-ylthio ) -4- ( 4-methoxyphenyl ) -4H-1,2,4-triazole-3-thiol ( 5j )The experimental process was repeated as with 4j ( 0.01 mole, 4.76 g ) and 2N NaOH and 2N HCl. The physical information is presented in Table 1.IR ( cm-1 ) : 1512 ( C=N ) ; 1H-NMR ? : 13.76 ( s, 1H, SH ) , 7.02-7.55 ( m, 14H, Ar-H ) , 3.82 ( s, 3H, OCH3 ) ; 13C-NMR ? : 168.65 ( C3 of triazole ) , 150.23 ( C5 of triazole ) , 131.74 ( C1 of phenyl ring ) , 130.78 ( C2 and C6 of phenyl ring ) , 129.72 ( C3 and C5 of phenyl ring ) , 128.67 ( C4 of phenyl ring ) , 55.29 ( -OCH3 ) ; MS m/z: 458.Synthesis of 5- ( 4,5-diphenyl-4H-1,2,4-triazol-3-ylthio ) -4- ( 4-ethoxyphenyl ) -4H-1,2,4-triazole-3-thiol ( 5k )The experimental process was repeated as with 4k ( 0.01 mole, 4.90 g ) and 2N NaOH and 2N HCl. The physical information is presented in Table 1.IR ( cm-1 ) : 1498 ( C=N ) ; 1H-NMR ? : 14.12 ( s, 1H, SH ) , 7.18-7.63 ( m, 14H, Ar-H ) , 3.67 ( Q, 2H, OCH2 ) , 1.54 ( T, 3H, CH3 ) ; 13C-NMR ? : 168.34 ( C3 of triazole ) , 150.33 ( C5 of triazole ) , 132.63 ( C1 of phenyl ring ) , 131.43 ( C2 and C6 of phenyl ring ) , 129.67 ( C3 and C5 of phenyl ring ) , 128.35 ( C4 of phenyl ring ) , 64.36 ( -CH2 ) , 14.21 ( -CH3 ) ; MS m/z: 472.Synthesis of 5- ( 4,5-diphenyl-4H-1,2,4-triazol-3-ylthio ) -4-isopropyl-4H-1,2,4-triazole-3-thiol ( 5l )The experimental process was repeated as with 4l ( 0.01 mole, 4.12 g ) and 2N NaOH and 2N HCl. The physical information is presented in Table 1.IR ( cm-1 ) : 1509 ( C=N ) ; 1H-NMR ? : 13.72 ( s, 1H, SH ) , 7.28-7.56 ( m, 10H, Ar-H ) , 4.12 ( m, 1H, CH ) , 1.57 ( vitamin D, 6H, 2CH3 ) ; 13C-NMR ? : 168.83 ( C3 of triazole ) , 150.84 ( C5 of triazole ) , 132.74 ( C1 of phenyl ring ) , 131.83 ( C2 and C6 of phenyl ring ) , 129.12 ( C3 and C5 of phenyl ring ) , 128.78 ( C4 of phenyl ring ) , 32.82 ( CH ) , 24.45 ( CH3 ) ; MS m/z: 394.Biological activityIn vitro antioxidant activityIn vitro antioxidant activity was carried out by DPPH extremist scavenging check method ( Roopan et al 2008, Yukesk et al 2006 ) . A stock solution of Ascorbic acid was prepared by fade outing 10mg in 10ml of Methanol from this a consecutive dilution of concentration 20, 40, 60, 80 and 100 µg/ml were prepared. Similarly A stock solution of Synthesized compounds were prepared by fade outing 10mg in 10ml of Methanol from this a consecutive dilutions of concentration 20, 40, 60, 80 and 100 µg/ml were prepared. The check was carried out utilizing UV- spectrophotometer at 517 nanometer. To 1ml of assorted concentrations of synthesised compounds, 1ml DPPH solution ( 40µg/ml ) was added into the trial tubing. The solution was incubated at 37 0C for 30 min and the optical density of each solution was measured at 517 nanometer against the corresponding trial and standard spaces. Experiment was performed in triplicate. The per centum free extremist scavenging activity can be calculated by the expression given below.Control – Trial% Scavenging = — — — — — — — — — — X 100ControlIn vitro cytotoxicity activityIn vitro cytotoxicity activities of the synthesized compounds against three human malignant neoplastic disease cell lines were HT-29 ( glandular cancer ) , EAC ( ehrich ascites carcinoma ) and MDA-231 ( Breast malignant neoplastic disease ) . The cell lines were procured from National Centre for Cell Sciences, Pune, India, were cultured in DMEM medium supplemented with 10 % FBS, 1 % L-glutamine, and 50 µg/ml Garamycin sulphate in a CO2 brooder in a humidified ambiance of 5 % CO2 and 95 % air. The in vitro cytotoxicity was determined utilizing a standard MTT check ( Molinari et al. , 2009, Manjula et al. , 2009, Purohit et al 2010 ) . Briefly, the exponentially turning cells were plated in 96-well home bases ( 104 cells/well in 100 µl of medium ) and incubated for 24 H for fond regard. The trial compounds were prepared prior to the survey by fade outing in 0.1 % DMSO and diluted with medium. The cells were so exposed to different concentration of trial compounds ( 200, 100, 50, 25 and 10 µg/ml ) in a volume of 100 µl/well. The cells in the growing control wells received merely the same volume of medium incorporating 0.1 % DMSO. After 72 H of exposure, the medium was removed and the cell civilizations were incubated with 100 µl of MTT reagent ( 0.1 % ) for 4 H at 370C. The pink colored formazan was dissolved in 100 µl of DMSO and optical density of each well was read in an ELISA micro home base reader at 570 nanometer. The experiment was performed in triplicate and the per centum cytotoxicity was calculated utilizing the undermentioned expression.The drug concentration that causes 50 % cell growing suppression after 72 H of uninterrupted exposure to the trial compounds ( IC50 ) was determined by plotting the graph of concentration of the drug against the per centum cytotoxicity and executing the arrested development analysis. The IC50 values of the trial compounds are shown in Table 3.In vitro antibacterial activitySynthesized compounds were evaluated for in vitro antibacterial activity at 100mg/ml concentration by cup home base method ( Rostom et al 2009. , Hugo et al 1997 ) . The trial beings used for antibacterial activity were Staphylococcus aureus, Bacillus substilus ( gms positive ) and Pseudomonas aeruginosa, Escherichia coli ( grams negative ) .Test compounds and mention criterions were dissolved in DMSO and sterile distilled H2O, severally, for the readying of stock and on the job solutions. Sterile alimentary agar home bases were prepared by pouring the unfertile agar into Petri dishes in sterile conditions. Overnight civilizations of bacteriums beings were adjusted to 106 c.f.u. ml-1 harmonizing to the Mac-Farland turbidness criterions. Each standardised trial being civilization ( 0.5ml ) was spread on to the agar home bases. Needed Numberss of pits were made by utilizing a unfertile bore bit of diameter 6 millimeter on the agar home bases. The solution incorporating 100?g/ml concentrations of the trial compounds and mention criterions were placed on the several pits. Cavity incorporating DMSO was used as solvent control. The home bases were maintained at 4 & A ; deg ; C for 1 H to let the diffusion of the solution into the medium and incubated at 37 & A ; deg ; C for 24h. After the incubation period, the zone of suppression was measured in millimeter.Recognitions Writers are grateful to The Principal, JSS College of Pharmacy, Mysore, India for supplying necessary installations. Writers besides thankful to The Director, NMR research Centre, Indian Institute of Science, Bangalore for spectral informations. Thankss are due to Mr. V.M. Chandrashekhar, Asst. Professor, Department of Pharmacology, HSK college of Pharmacy, Bagalkot, India for transporting out anti-cancer activity.

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