Integrated In Silico and In Vivo Toxicogenetic Assessment of Piperine and 4,5-Dihydroxypiperine in Drosophila melanogaster

Abstract 4,5-Dihydroxypiperine (dhPIP) is a synthetic derivative of piperine (PIP), a bioactive alkaloid investigated as a phytogenic additive. This study evaluated the in vitro antioxidant capacity, in silico pharmacokinetic, and in vivo toxicogenetics of PIP and dhPIP using standard (ST) and high-bioactivation (HB) crosses of Drosophila melanogaster. DhPIP exhibited higher antioxidant activity than PIP. In silico, high intestinal absorption and low blood–brain barrier permeability was predicted for both, but CYP450 interactions were exclusive to PIP. In vivo, neither compound was genotoxic. Notably, both alkaloids reduced doxorubicin-induced DNA damage. In the ST cross, PIP inhibited 92–98% of genotoxicity, dropping to 51–55% in the HB cross. Conversely, dhPIP reduced damage by approximately 50% in both crosses. These findings demonstrate that PIP and dhPIP lack genotoxic effects, highlighting that while dhPIP offers enhanced antioxidant capacity, its in vivo antigenotoxicity is lower than PIP’s but remains unaffected by CYP450-mediated metabolism.