Regio- and Stereoselective Synthesis of Nitro-fatty Acids as NRF2 Pathway Activators Working under Ambient or Hypoxic Conditions.

Nitro-fatty acids (NO(2)FAs) are endogenously produced electrophiles and NRF2 activators with therapeutic potential. We developed a synthetic protocol combining a Henry reaction and base-promoted β-elimination, yielding ultrapure regio/stereoisomers of nitro-stearic (NO(2)SA), nitro-oleic (NO(2)OA), and conjugated/bis-allylic nitro-linoleic (NO(2)LA) acids. These were tested for NRF2 pathway activation in bone marrow cells under different oxygen conditions. We observed that 9- and 10-NO(2)OA, and 10-NO(2)LA increased NRF2 stabilization under hypoxia, while 9- and 10-NO(2)OA significantly upregulated Hmox1 and Gclm at all oxygen levels. 9- and 10-NO(2)OA enhanced HO-1 and GCLM proteins independently of oxygen, while 10-NO(2)LA was oxygen-dependent, boosting HO-1 under hypoxia and GCLM under ambient conditions. Moreover, 10-NO(2)OA and 10-NO(2)LA induced NRF2 nuclear translocation. In contrast, the saturated 10-NO(2)SA, which has lower electron-acceptor ability, was inactive. In summary, these findings suggest the biological activity of NO(2)FAs is dependent on oxygen level, which could be used in future research of other oxidative stress-dependent pathways.