A strategy for liver selective NRF2 induction via cytochrome P450-activated prodrugs with low activity in hypoxia

Activation of the transcription factor nuclear factor-erythroid 2-related factor 2 (NRF2) has been shown to be a promising therapeutic approach in the treatment of hepatosteatosis. NRF2 is believed to exert beneficial effects by upregulating cellular oxidative defense mechanisms and inhibiting inflammation. However, a major concern associated with long-term treatment with NRF2 activators are drug side effects, including the promotion of tumorigenesis. Many NRF2 activators function by forming cysteine adducts with KEAP1, which normally mediates the ubiquitination and degradation of NRF2. In this study, we identified NRF2 activator prodrugs of 4-methylcatechol and tert-butylhydroquinone. These prodrugs are converted into their active metabolites in a liver selective, cytochrome P450-dependent manner and function by inhibiting KEAP1, resulting in NRF2 activation. Unexpectedly, we also found that a number of NRF2-activating compounds, including 4-methylcatechol and tert-butylhydroquinone, show a markedly lower activity under hypoxic conditions than normoxia. Our findings suggest that the lower activity of these NRF2 inducers is a consequence of less potent cysteine adduct formation with KEAP1. The lower activity of NRF2 inducing compounds in hypoxia may limit tumor promoting effects of NRF2 induction. Our study provides an important proof of concept that it is possible to selectively activate NRF2 in the liver for the treatment of hepatosteatosis while avoiding tumorigenic effects as well as side effects of NRF2 activation in other tissues.