In vitro analysis of the susceptibility of HIV-1 subtype A and CRF01_AE integrases to raltegravir

Int J Antimicrob Agents. 2014 Aug;44(2):168-72. doi: 10.1016/j.ijantimicag.2014.04.013. Epub 2014 Jun 2.

Abstract

The antiviral efficacy of raltegravir (RAL) has been proven against human immunodeficiency virus type 1 (HIV-1) subtypes B and C but remained to be determined against other subtypes. Therefore, the enzymatic activities as well as RAL resistance of HIV-1 subtype A and CRF01_AE integrases (INs) were investigated. Previously published subtype A and CRF01_AE IN sequences from RAL-naïve patients were aligned to generate consensus sequences for both IN subtypes. Subtype A and CRF01_AE INs encoded by these consensus sequences as well as the corresponding enzymes harbouring the N155H resistance mutation were expressed and purified. Enzymatic activities of subtype A and CRF01_AE INs were analysed with regard to typical 3'-end processing (3'-P) and strand transfer (ST) activities both in the presence and absence of RAL and were compared with subtype B IN as well as with the corresponding INs harbouring the N155H resistance mutation. Subtypes B, A and CRF01_AE INs showed similar 3'-P and ST activities. In the presence of RAL, the three wild-type INs exhibited ST activity IC50 values (50% inhibitory concentrations) of 86.3 ± 32.5, 158.3 ± 99.0 and 100.0 ± 65.7 nM, respectively. Analysis of 3'-P activity in the presence of RAL revealed IC(50) > 10 μM for all three enzymes. The three INs harbouring the N155H mutation presented in vitro low but similar resistance levels to RAL. In conclusion, INs from HIV-1 subtypes B, A and CRF01_AE showed similar responses to RAL in vitro, suggesting the potency of this antiretroviral drug to treat HIV-1 subtype A- and CRF01_AE-infected patients.

Keywords: CRF01_AE HIV-1; Inhibition; Integrase; Raltegravir; Subtype A HIV-1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-HIV Agents / pharmacology*
  • Cloning, Molecular
  • Gene Expression
  • Genotype
  • HIV Integrase / genetics
  • HIV Integrase / metabolism*
  • HIV-1 / classification
  • HIV-1 / drug effects*
  • HIV-1 / genetics
  • HIV-1 / isolation & purification
  • Humans
  • Inhibitory Concentration 50
  • Microbial Sensitivity Tests
  • Mutant Proteins / genetics
  • Mutant Proteins / metabolism
  • Pyrrolidinones / pharmacology*
  • Raltegravir Potassium

Substances

  • Anti-HIV Agents
  • Mutant Proteins
  • Pyrrolidinones
  • Raltegravir Potassium
  • HIV Integrase
  • p31 integrase protein, Human immunodeficiency virus 1