Nrf2/HO-1 activator 2 (compound 13m), a difluoro-substituted derivative, is a highly potent activator of Nrf2/HO-1 with substantial neuroprotective and antioxidant properties. It functions by activating the Nrf2/HO-1 pathway through the phosphorylation of ERK1/2, JNK, or Akt in PC12 cells and is useful in investigating Parkinson's disease (PD) [1].

Nrf2/HO-1 activator 2, at concentrations ranging from 0.1-30 μM for 24 hours, offers protective effects to PC12 cells against 6-OHDA- and Rotenone-induced toxicity and cell death. At 1-100 μM for 11 hours, it inhibits reactive oxygen species production and partially reduces lipid peroxidation in rat brain homogenates, decreasing lipid peroxide production by 28.8% at 100 μM. Additionally, it promotes up-regulation of heme oxygenase-1 (HO-1) and Nrf2 expression and activates Nrf2/HO-1 signaling pathways through phosphorylation of ERK1/2, JNK, and Akt in PC12 cells. Cell viability assays demonstrate its neuroprotective effects, rescuing cells from rotenone-induced damage and improving cell viability up to 79.9% at 30 μM. Western blot analysis further confirms dose-dependent upregulation of HO-1 levels and enhanced nuclear translocation of Nrf2, as well as inducing phosphorylation of ERK1/2, JNK, and Akt at 10 μM.