S. N. Mokale et al. / Bioorg. Med. Chem. 20 (2012) 3119–3127
3127
4.3.13. 4-(2,4-Dichlorobenzylidene)-2-phenyl-1-(pyridin-4-yl
1H NMR d: 8.6 (s, 2H), 7.8–7.5 (m, 12H), 4.5 (s, 2H), 3.5 (s, 3H);
MS: m/z 370 (M+1).
methyl)-1H-imidazol-5(4H)-one (4m)
IR (KBr): 3063, 2986, 1754, 1582, 1651, 1446, 865 cmꢀ1
1H NMR: d 8.4 (s, 2H), 7.5–6.8 (m, 11H), 4.7 (s, 2H);
MS: m/z 409(M+1);
;
4.4. In vitro anti-HIV activity
The standard Nevirapine and test compounds (4a–t) were pow-
4.3.14. 4-(4-(Dimethylamino) benzylidene)-2-phenyl-1-(pyridin
dered finely and suspended in DMSO. Finally 20 lg sample was
-4-ylmethyl)-1H-imidazol-5(4H)-one (4n)
used for in vitro assay. The RT inhibition assay was performed by
using an RT assay kit (Roche). The procedure for assaying RT inhi-
bition was performed as described in the kit protocol. Briefly, the
reaction mixture consists of template primer complex, dNTPs and
reverse transcriptase (RT) enzyme in the lysis buffer with or with-
out inhibitors. After 1-h incubation at 37 °C, the reaction mixture
was transferred to streptavidine-coated microtitre plate (MTP).
The biotin-labeled dNTPs that are incorporated in the template
due to activity of RT were bound to streptavidine. The unbound
dNTPs were washed using wash buffer and anti-DIG-POD was
added to the MTP. The DIG-labeled dNTPs incorporated in the tem-
plate were bound to anti-DIG-POD Antibody. The unbound anti-
DIG-POD was washed and the peroxide substrate (ABST) was
added to the MTP. A colored reaction product was produced during
the cleavage of the substrate catalyzed by a peroxide enzyme. The
absorbance of the sample was determined at O.D. Four hundred
and five nanometers using micro titer plate ELISA reader. The %
inhibition was calculated using following formula.
IR (KBr): 3052, 2920, 1725,1606,1648,1449,891 cmꢀ1
;
1H NMR: d 8.6(s, 2H), 7.5–6.8 (m, 12H), 4.5(s, 2H), 3.8 (s, 6H);
MS: m/z 383(M+1);
4.3.15. 4-(2-Hydroxybenzylidene)-2-phenyl-1-(pyridin-4-yl
methyl)-1H-imidazol-5(4H)-one (4o)
IR (KBr): 3563, 3055, 2962, 1746, 1650, 1604, 1443, 878 cmꢀ1
1H NMR: d 8.5 (s, 2H), 8.1–7.8 (m, 12H), 5.5 (s, 1H), 4.8 (s, 2H);
M.S: m/z 3356 (M+1);
;
4.3.16. 4-(Furan-2-ylmethylene)-2-phenyl-1-(pyridin-4-yl
methyl)-1H-imidazol-5(4H)-one (4p)
IR (KBr): 3055, 2962, 1746, 1650, 1604, 1443, 878 cmꢀ1
1H NMR: d 8.5 (s, 2H), 8.1–7.8 (m, 11H), 4.5 (s, 2H);
M.S: m/z 330 (M+1);
;
4.3.17. 4-(4-Hydroxy-3-methoxybenzylidene)-2-phenyl-1-(pyri
din-4-ylmethyl)-1H-imidazol-5(4H)-one (4q)
ꢀ
ꢁ
OD at 405nm with inhibitor
IR (KBr): 3563, 3052, 1734, 1650, 1602, 1443, 1381, 884 cmꢀ1
;
% inhibition ¼ 100 ꢀ
ꢁ 100
OD at 405nm without inhibitor
1H NMR: d 8.5 (s, 2H), 7.8–7.5 (m, 12H), 5.5 (s, 1H), 4.8 (s, 2H),
3.8 (s, 3H);
MS: m/z 386 (M+1);
References and notes
1. Balzarini, J.; Rao, A.; Carbone, A. Antiviral Res. 2004, 63, 79.
2. Rawal, R.; Tripathi, R.; Katti, S. B. Eur. J. Med. Chem. 2008, 43, 2800.
4.3.18. 4-Ethylidene-2-phenyl-1-(pyridin-4-ylmethyl)-1H-imida
zol-5(4H)-one (4r)
3. Loksha, Y. M.; El-Barbary, A. A.; Nielsen, C. Bioorg. Med. Chem. 2005, 13, 4209.
4. Charles, E.; Mowbray, A.; Burt, C. Bioorg. Med. Chem. Lett. 2009, 19, 5599.
5. Gagnon, A.; Jakalian, A.; Guse, I. Bioorg. Med. Chem. Lett. 2009, 19, 1199.
6. Zhan, P.; Liu, X.; Fang, Z. Eur. J. Med. Chem. 2009, 44, 4648.
7. Masuda, N.; Yamamoto, O. Bioorg. Med. Chem. Lett. 2005, 13, 949.
8. Wu, J.; Liu, X. Molecules 2007, 12, 2003.
IR (KBr): 2984, 2877, 1735, 1632, 1585, 1441, 884 cmꢀ1
;
1H NMR: d 8 .7 (s, 2H), 7.8–7.3 (m, 8H), 4.6 (s, 2H), 2.3 (s, 3H);
MS: m/z 278 (M+1);
9. Rawal, R.; Tripathi, R.; Katti, S. B. Bioorg. Med. Chem. 2007, 15, 1725.
10. Friesner, R. A.; Murphy, R. B.; Repasky, M. P.; Frye, L. L.; Greenwood, J. R.;
Halgren, T. A.; Sanschagrin, P. C.; Mainz, D. T. J. Med. Chem. 2006, 49, 6177.
11. Wallace, A. C.; Laskowski, R. A.; Thornton, J. M. Protein Eng. 1995, 8, 127.
12. Pawar, V.; Lokwani, D.; Bhandari, S.; Mitra, D.; Sabde, S.; Bothara, K.;
Madgulkar, A. Bioorg. Med. Chem. 2010, 18, 3198.
4.3.19. 2-Phenyl-1-(pyridin-4-ylmethyl)-4-(3,4,5-trimethoxy
benzylidene)-1H-imidazol-5(4H)-one (4s)
IR (KBr): 3004, 2959, 1715, 1603, 1650, 1443, 888 cmꢀ1
;
1H NMR d: 8.6 (s, 2H), 7.8–7.5 (m, 10H), 4.5 (s, 2H), 3.5 (s, 9H);
MS: m/z 430 (M+1);
13. Furniss, B. S. Handbook of Practical Organic Chemistry, 5th ed.; Pearson
education, 2008. p 1155.
4.3.20. 4-(4-Methoxybenzylidene)-2-phenyl-1-(pyridin-4-yl
methyl)-1H-imidazol-5(4H)-one (4t)
IR (KBr): 3023, 2917, 1730, 1575, 1646, 1447, 863 cmꢀ1
;