Covalent Histone Modification by an Electrophilic Derivative of the Anti-HIV Drug Nevirapine

Abstract

Nevirapine (NVP), a non-nucleoside reverse transcriptase inhibitor widely used in combined antiretroviral therapy and to prevent mother-to-child transmission of the human immunodeficiency virus type 1, is associated with several adverse side effects. Using 12-mesyloxy-nevirapine, a model electrophile of the reactive metabolites derived from the NVP Phase I metabolite, 12-hydroxy-NVP, we demonstrate that the nucleophilic core and C-terminal residues of histones are targets for covalent adduct formation. We identified multiple NVP-modification sites at lysine (e.g., H2BK47, H4K32), histidine (e.g., H2BH110, H4H76), and serine (e.g., H2BS33) residues of the four histones using a mass spectrometry-based bottom-up proteomic analysis. In particular, H2BK47, H2BH110, H2AH83, and H4H76 were found to be potential hot spots for NVP incorporation. Notably, a remarkable selectivity to the imidazole ring of histidine was observed, with modification by NVP detected in three out of the 11 histidine residues of histones. This suggests that NVP-modified histidine residues of histones are prospective markers of the drug’s bioactivation and/or toxicity. Importantly, NVP-derived modifications were identified at sites known to determine chromatin structure (e.g., H4H76) or that can undergo multiple types of post-translational modifications (e.g., H2BK47, H4H76). These results open new insights into the molecular mechanisms of drug-induced adverse reactions.

Synthesis and characterization of two known and one new impurities of dolutegravir: In silico evaluation of certain intermediates against SARS CoV-2 O-ribose methyltransferase (OMTase)

“…With over 37 million people currently being affected worldwide [ 1 , 2 , 3 ] by HIV, the disease remained as one of the major health concerns. While a range of treatments have been introduced to control this disease [ 4 , 5 ] however the high cost often poses an economic burden for patients especially in countries belonging to lower-income group. Launched in the market in 2013 under the umbrella of second generation integrase inhibitors [6] the anti-HIV drug dolutegravir (GSK1349572; brand name ‘Tivicay’) was developed by GlaxoSmithKline (GSK) and Shionogi.…”

Bottom-up proteomics analysis for adduction of the broad-spectrum herbicide atrazine to histone

“…Histones affect most DNA-related events, including transcription, replication, and chromosome segregation, and play a key role in controlling DNA expression [ 14 , 17 – 21 ]. Recent evidence has shown that alteration of histones by exogenous toxicants might conceivably disturb their normal function [ 14 , 19 , 22 – 25 ].…”

Asparagine and Glutamine Residues Participate in Protein Covalent Binding by Epoxide Metabolite of 8-Epidiosbulbin E Acetate In Vitro and In Vivo

“…Nucleophilic amino residues, such as lysine and cysteine, of proteins are the major modification targets of electrophilic metabolites and intermediates. , Lysine residues play a key role in maintaining normal physiological functions of many proteins. , The primary amine group of lysine in the protein peptide chain is a nucleophilic group that is easily attacked by electrophiles, in particular, those hard electrophiles, to form modified proteins, and these modifications are often associated with chemical-induced toxicity. , Our previous work demonstrated EEA was metabolized to cis -enedial 3 (Scheme ) which sequentially reacted with lysine residues of proteins to form pyrroline derivative 8 (Scheme ), the structure of which was confirmed by NMR . The EEA-derived protein adductions were verified by rationally designed polyclonal antibodies developed in our laboratory …”