A series of 2′,3′-exo-methylene carbocyclic nucleosides was synthesized as potential antiviral agents. These compounds were built on a bicyclo[3.1.0]hexane template that exhibits a rigid pseudoboat conformation and is capable of maintaining an identical conformation in solid state and in solution. The structures of the synthesized compounds were elucidated by NMR and X-ray crystallography. All the compounds were tested as anti-HIV and anti-HSV agents. The chemically synthesized 5′-triphosphate analogue of 7 was evaluated directly as a reverse transcriptase inhibitor.
Bibliographical noteFunding Information:
Purified recombinant HIV-1 reverse transcriptase was obtained from the University of Alabama at Birmingham, Center for AIDS Research, Gene Expression Core Facility (supported in part by the NIH Centers for AIDS research program grant P30 AI27767). For the study of RNA transcription, HIV-1 reverse transcriptase activity was measured in 50-μL reactions containing 50 mM Tris (pH 8.0), 50 mM KCl, 10 mM MgCl 2 , 4 mM β-mercaptoethanol, 3% glycerol, 1 mg/mL bovine serum albumin, 3.33 μg/mL of primed 16S rRNA from Escherichia coli, 10 μM dTTP, 10 μM dGTP, 10 μM dCTP, and 0.25 μM [ 33 P]dATP (the K m concentration). The primer was annealed to the template at a ratio of 3 to 1 as described earlier. 14 After incubation the DNA in each sample was precipitated onto glass fiber filters using a 5% trichloroacetic acid solution containing 10 mM pyrophosphate. These filters were batch washed and counted for radioactivity. 15 Assays were done in duplicate and the IC 50 values were reported.
Copyright 2008 Elsevier B.V., All rights reserved.