Effect of ethanol on spectral binding, inhibition, and activity of CYP3A4 with an antiretroviral drug nelfinavir

Santosh Kumar, Ravinder Earla, Mengyao Jin, Ashim K. Mitra, Anil Kumar

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Cytochrome P450 3A4 (CYP3A4) is the most abundant CYP enzyme in the liver and metabolizes approximately 50% of the drugs, including antiretrovirals. Although CYP3A4 induction by ethanol and impact of CYP3A4 on drug metabolism and toxicity is known, CYP3A4-ethanol physical interaction and its impact on drug binding, inhibition, or metabolism is not known. Therefore, we studied the effect of ethanol on binding and inhibition of CYP3A4 with a representative protease inhibitor, nelfinavir, followed by the effect of alcohol on nelfinavir metabolism. Our initial results showed that methanol, ethanol, isopropanol, isobutanol, and isoamyl alcohol bind in the active site of CYP3A4 and exhibit type I spectra. Among these alcohol compounds, ethanol showed the lowest KD (5.9±0.34mM), suggesting its strong binding affinity with CYP3A4. Ethanol (20mM) decreased the KD of nelfinavir by >5-fold (0.041±0.007 vs. 0.227±0.038μM). Similarly, 20mM ethanol decreased the IC50 of nelfinavir by >3-fold (2.6±0.5 vs. 8.3±3.1μM). These results suggest that ethanol facilitates binding of nelfinavir with CYP3A4. Furthermore, we performed nelfinavir metabolism using LCMS. Although ethanol did not alter kcat, it decreased the Km of nelfinavir, suggesting a decrease in catalytic efficiency (kcat/Km). This is an important finding because alcoholism is prevalent in HIV-1-infected persons and alcohol is shown to decrease the response to antiretroviral therapy.

Original languageEnglish (US)
Pages (from-to)163-167
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume402
Issue number1
DOIs
StatePublished - Nov 5 2010
Externally publishedYes

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Nelfinavir
Cytochrome P-450 CYP3A
Ethanol
Pharmaceutical Preparations
Metabolism
Alcohols
Enzyme inhibition
Inhibition (Psychology)
2-Propanol
Protease Inhibitors
Drug-Related Side Effects and Adverse Reactions
Liver
Alcoholism
Inhibitory Concentration 50
Methanol
Toxicity
HIV-1
Catalytic Domain
Efficiency

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Effect of ethanol on spectral binding, inhibition, and activity of CYP3A4 with an antiretroviral drug nelfinavir. / Kumar, Santosh; Earla, Ravinder; Jin, Mengyao; Mitra, Ashim K.; Kumar, Anil.

In: Biochemical and Biophysical Research Communications, Vol. 402, No. 1, 05.11.2010, p. 163-167.

Research output: Contribution to journalArticle

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