Biotransformation of efavirenz and proteomic analysis of P450s and UGTs in human, mouse, and macaque brain-derived in vitro systems

ABSTRACT: Antiretroviral drugs such as efavirenz (EFV) are essential to combat HIV infection in the brain, but little is known about how these drugs are metabolized locally. In this study, the cytochrome P450 (P450) and UDP-glucuronosyltransferase (UGT)-dependent metabolism of EFV was probed in brain microsomes from mice, cynomolgus macaques, and humans as well as primary neural cells from C57BL/6N mice. Utilizing ultra-high performance liquid chromatography high resolution mass spectrometry (uHPLC-HRMS), the formation of 8-hydroxyefavirenz (8-OHEFV) from EFV and the glucuronidation of P450-dependent metabolites 8-OHEFV and 8,14-dihydroxyefavirenz (8,14-diOHEFV) was observed in brain microsomes from all three species. The direct glucuronidation of EFV, however, was only detected in the cynomolgus macaque brain microsomes. In brain cell typesprimary neural cells treated with EFV, microglia formed 8-OHEFV only while glucuronidation was detected in cortical neurons and astrocytes treated with P450-dependent metabolites of EFV. Untargeted and targeted proteomics experiments were used to determine which P450s and UGTs were present in the brain microsomes. Eleven P450s and 11 UGTs were detected in the human brain microsomes, whilewith 10 P450s and 15 UGTs were identified in the mouse brain microsomes and 17 and 6 P450s and UGTs, respectively, observed in the macaque brain microsomes. For many of these enzymes, this was the first time they have been noted at the protein level in the brain microsomes. This study indicates the potential for the brain to play a contributing role in the local metabolism of EFV, and the related pharmacological and toxicological consequences.metabolism to contribute to pharmacological and toxicological EFV outcomes in brain.

INSTRUMENT(S): ZenoTOF 7600

ORGANISM(S): 9606

SUBMITTER: Benjamin C Orsburn  

PROVIDER: MSV000090576 | MassIVE | Mon Oct 24 10:11:00 BST 2022

REPOSITORIES: MassIVE