Effects of Diterpenes Isolated from the Brazilian Marine Alga Dictyota menstrualis on HIV-1 Reverse Transcriptase

Abstract

It has been recently demonstrated that HIV-1 reverse transcriptase is the target of two diterpenes, (6R)-6-hydroxydichotoma-3,14-diene-1,17-dial (compound 1) and (6R)-6-acetoxydichotoma-3,14-diene-1,17-dial (compound 2), that inhibit HIV-1 replication in vitro. In this work, the effects of both diterpenes on the kinetic properties of the recombinant HIV-1 reverse transcriptase (RT) enzyme were evaluated. RNA-dependent DNA-polymerase (RDDP) activity assays demonstrated that both diterpenes behave as non-competitive inhibitors with respect to dTTP and uncompetitive inhibitors with respect to poly(rA)·oligo(dT) template primers. The K _i values obtained for compounds 1 and 2 were 10 and 35 μM, respectively. Neither of these diterpenes affected the DNA-dependent DNA-polymerase (DDDP) activity of the HIV-1 RT. The RDDP activities of AMV-RT and MMLV-RT enzymes were also inhibited by compounds 1 and 2. In contrast to the HIV-1 enzyme, the DDDP activities of AMV-RT and MMLV-RT enzymes were significantly reduced by compound 1. Taken together, our results demonstrate that compound 1 is a more effective inhibitor of the viral reverse transcriptases from HIV-1, AMV and MMLV than compound 2. The kinetic behavior analyses of the HIV-1 RT demonstrate that both diterpenes have similar mechanisms of inhibition of RDDP activity.

Abbreviations

Compound 1:(6R)-6-hydroxydichotoma-3,14-diene-1,17-dial

Compound 2:(6R)-6-acetoxydichotoma-3,14-diene-1,17-dial

HIV-1 RT:human immunodeficiency virus type-1 reversetranscriptase

AMV:avian myeloblastosis virus

MMLV:Moloney murine leukemia virus

Key words

Diterpenes - HIV-1 RT - AMV-RT - MMLV-RT - non-competitive inhibitors - uncompetitive inhibitors

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