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02442 PMPA- NNRTI COMBINATION STUDIES DEMONSTRATE POTENT SYNERGISM AGAINST HIV-1 INFECTION IN VITRO Schader, Susan* Background: Combination products may maximise microbicide efficacy through: blockade of multiple targets involved in HIV transmission; increased activity through potential synergy; and minimizing potential development of resistance. We have investigated the synergistic potential of PMPA (9-[2-(Phosphonomethoxy)Propyl] Adenine), an acyclic nucleoside phosphonate (NRTI), with non- nucleoside reverse transcriptase inhibitors (NNRTIs) and PRO 2000, a candidate polyanion microbicide. Methods: Synergistic potential was evaluated in cellular and cervical explant studies. Cells or tissue were treated with each drug alone or in a fixed ratio combination for 1 hour. R5 or X4 HIV-1 isolates were then added and incubated for a further 2 hours with subsequent washing to remove free drug and virus. Viral replication was monitored by measurement of supernatant reverse transcriptase activity (RT), p24 release or quantitative proviral PCR. The 50% inhibitory concentration (IC50) for drugs alone and in combination was determined. Analysis of combined effects was accomplished using the median effect principle developed by Chou and Talalay, (Calcusyn, Biosoft). Results: PMPA in combination with UC781 demonstrated anti- HIV- 1 synergism with combination indices <1 in RT based assays, while PMPA in combination with PRO 2000 did not. These results have been extended in a cervical explant model to evaluate the efficacy of single and combination drugs against HIV infection and potential dissemination by migratory cells. Conclusions: PMPA, in combination with candidate NNRTIs demonstrate good anti-HIV-1 activity in vitro and the capability of these drugs to demonstrate synergy is promising. Although PRO 2000 in combination with PMPA might improve the effect of PRO 2000, the combination is not synergistic and would not enhance the efficacy of PMPA. Ms. Susan Schader- Plesman |
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