Differential Effects of Ethanol on Spectral Binding and Inhibition of Cytochrome P450 3A4 with Eight Protease Inhibitors Antiretroviral Drugs

Santosh Kumar, Anil Kumar

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Background: Cytochrome P450 3A4 (CYP3A4) is the most abundant CYP enzyme in the liver, which metabolizes approximately 50% of the marketed drugs including antiretroviral agents. CYP3A4 induction by ethanol and its impact on drug metabolism and toxicity is known. However, CYP3A4-ethanol physical interaction and its impact on drug binding, inhibition, or metabolism is not known, except that we have recently shown that ethanol facilitates the binding of a protease inhibitor (PI), nelfinavir, with CYP3A4. The current study was designed to examine the effect of ethanol on spectral binding and inhibition of CYP3A4 with all currently used PIs that differ in physicochemical properties. Methods: We performed type I and type II spectral binding with CYP3A4 at 0 and 20mM ethanol and varying PIs' concentrations. We also performed CYP3A4 inhibition using 7-benzyloxy-4-trifluoromethylcoumarin substrate and NADPH at varying concentrations of PIs and ethanol. Results:  Atazanavir, lopinavir, saquinavir, and tipranavir showed type I spectral binding, whereas indinavir and ritonavir showed type II. However, amprenavir and darunavir did not show spectral binding with CYP3A4. Ethanol at 20mM decreased the maximum spectral change (δA max) with type I lopinavir and saquinavir, but it did not alter δA max with other PIs. Ethanol did not alter spectral binding affinity (K D) and inhibition constant (IC 50) of type I PIs. However, ethanol significantly decreased the IC 50 of type II PIs, indinavir and ritonavir, and markedly increased the IC 50 of amprenavir and darunavir. Conclusions: Overall, our results suggest that ethanol differentially alters the binding and inhibition of CYP3A4 with the PIs that have different physicochemical properties. This study has clinical relevance because alcohol has been shown to alter the response to antiretroviral drugs, including PIs, in HIV-1-infected individuals.

Original languageEnglish (US)
Pages (from-to)2121-2127
Number of pages7
JournalAlcoholism: Clinical and Experimental Research
Volume35
Issue number12
DOIs
StatePublished - Dec 1 2011
Externally publishedYes

Fingerprint

Cytochrome P-450 CYP3A
Protease Inhibitors
Ethanol
Pharmaceutical Preparations
Saquinavir
Lopinavir
Indinavir
Ritonavir
Metabolism
Nelfinavir
Anti-Retroviral Agents
Enzyme inhibition
Drug-Related Side Effects and Adverse Reactions
NADP
Liver
Toxicity
HIV-1
Alcohols

All Science Journal Classification (ASJC) codes

  • Medicine (miscellaneous)
  • Toxicology
  • Psychiatry and Mental health

Cite this

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title = "Differential Effects of Ethanol on Spectral Binding and Inhibition of Cytochrome P450 3A4 with Eight Protease Inhibitors Antiretroviral Drugs",
abstract = "Background: Cytochrome P450 3A4 (CYP3A4) is the most abundant CYP enzyme in the liver, which metabolizes approximately 50{\%} of the marketed drugs including antiretroviral agents. CYP3A4 induction by ethanol and its impact on drug metabolism and toxicity is known. However, CYP3A4-ethanol physical interaction and its impact on drug binding, inhibition, or metabolism is not known, except that we have recently shown that ethanol facilitates the binding of a protease inhibitor (PI), nelfinavir, with CYP3A4. The current study was designed to examine the effect of ethanol on spectral binding and inhibition of CYP3A4 with all currently used PIs that differ in physicochemical properties. Methods: We performed type I and type II spectral binding with CYP3A4 at 0 and 20mM ethanol and varying PIs' concentrations. We also performed CYP3A4 inhibition using 7-benzyloxy-4-trifluoromethylcoumarin substrate and NADPH at varying concentrations of PIs and ethanol. Results:  Atazanavir, lopinavir, saquinavir, and tipranavir showed type I spectral binding, whereas indinavir and ritonavir showed type II. However, amprenavir and darunavir did not show spectral binding with CYP3A4. Ethanol at 20mM decreased the maximum spectral change (δA max) with type I lopinavir and saquinavir, but it did not alter δA max with other PIs. Ethanol did not alter spectral binding affinity (K D) and inhibition constant (IC 50) of type I PIs. However, ethanol significantly decreased the IC 50 of type II PIs, indinavir and ritonavir, and markedly increased the IC 50 of amprenavir and darunavir. Conclusions: Overall, our results suggest that ethanol differentially alters the binding and inhibition of CYP3A4 with the PIs that have different physicochemical properties. This study has clinical relevance because alcohol has been shown to alter the response to antiretroviral drugs, including PIs, in HIV-1-infected individuals.",
author = "Santosh Kumar and Anil Kumar",
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doi = "10.1111/j.1530-0277.2011.01575.x",
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T1 - Differential Effects of Ethanol on Spectral Binding and Inhibition of Cytochrome P450 3A4 with Eight Protease Inhibitors Antiretroviral Drugs

AU - Kumar, Santosh

AU - Kumar, Anil

PY - 2011/12/1

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N2 - Background: Cytochrome P450 3A4 (CYP3A4) is the most abundant CYP enzyme in the liver, which metabolizes approximately 50% of the marketed drugs including antiretroviral agents. CYP3A4 induction by ethanol and its impact on drug metabolism and toxicity is known. However, CYP3A4-ethanol physical interaction and its impact on drug binding, inhibition, or metabolism is not known, except that we have recently shown that ethanol facilitates the binding of a protease inhibitor (PI), nelfinavir, with CYP3A4. The current study was designed to examine the effect of ethanol on spectral binding and inhibition of CYP3A4 with all currently used PIs that differ in physicochemical properties. Methods: We performed type I and type II spectral binding with CYP3A4 at 0 and 20mM ethanol and varying PIs' concentrations. We also performed CYP3A4 inhibition using 7-benzyloxy-4-trifluoromethylcoumarin substrate and NADPH at varying concentrations of PIs and ethanol. Results:  Atazanavir, lopinavir, saquinavir, and tipranavir showed type I spectral binding, whereas indinavir and ritonavir showed type II. However, amprenavir and darunavir did not show spectral binding with CYP3A4. Ethanol at 20mM decreased the maximum spectral change (δA max) with type I lopinavir and saquinavir, but it did not alter δA max with other PIs. Ethanol did not alter spectral binding affinity (K D) and inhibition constant (IC 50) of type I PIs. However, ethanol significantly decreased the IC 50 of type II PIs, indinavir and ritonavir, and markedly increased the IC 50 of amprenavir and darunavir. Conclusions: Overall, our results suggest that ethanol differentially alters the binding and inhibition of CYP3A4 with the PIs that have different physicochemical properties. This study has clinical relevance because alcohol has been shown to alter the response to antiretroviral drugs, including PIs, in HIV-1-infected individuals.

AB - Background: Cytochrome P450 3A4 (CYP3A4) is the most abundant CYP enzyme in the liver, which metabolizes approximately 50% of the marketed drugs including antiretroviral agents. CYP3A4 induction by ethanol and its impact on drug metabolism and toxicity is known. However, CYP3A4-ethanol physical interaction and its impact on drug binding, inhibition, or metabolism is not known, except that we have recently shown that ethanol facilitates the binding of a protease inhibitor (PI), nelfinavir, with CYP3A4. The current study was designed to examine the effect of ethanol on spectral binding and inhibition of CYP3A4 with all currently used PIs that differ in physicochemical properties. Methods: We performed type I and type II spectral binding with CYP3A4 at 0 and 20mM ethanol and varying PIs' concentrations. We also performed CYP3A4 inhibition using 7-benzyloxy-4-trifluoromethylcoumarin substrate and NADPH at varying concentrations of PIs and ethanol. Results:  Atazanavir, lopinavir, saquinavir, and tipranavir showed type I spectral binding, whereas indinavir and ritonavir showed type II. However, amprenavir and darunavir did not show spectral binding with CYP3A4. Ethanol at 20mM decreased the maximum spectral change (δA max) with type I lopinavir and saquinavir, but it did not alter δA max with other PIs. Ethanol did not alter spectral binding affinity (K D) and inhibition constant (IC 50) of type I PIs. However, ethanol significantly decreased the IC 50 of type II PIs, indinavir and ritonavir, and markedly increased the IC 50 of amprenavir and darunavir. Conclusions: Overall, our results suggest that ethanol differentially alters the binding and inhibition of CYP3A4 with the PIs that have different physicochemical properties. This study has clinical relevance because alcohol has been shown to alter the response to antiretroviral drugs, including PIs, in HIV-1-infected individuals.

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JF - Alcoholism: Clinical and Experimental Research

SN - 0145-6008

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