Pregnenolone carbonitrile is primarily used in research settings to study the regulation of cytochrome P450 enzymes, particularly CYP2B and CYP3A families. It acts as an inducer of these enzymes, making it valuable for understanding drug metabolism and the mechanisms of enzyme induction. In toxicology studies, it helps investigate the effects of enzyme induction on chemical detoxification processes. Additionally, it has been utilized in studies exploring steroidogenesis and the biosynthesis of steroid hormones, providing insights into endocrine system functions. Its role in modulating enzyme activity also makes it relevant in pharmacological research for evaluating drug interactions and metabolic pathways.