In vitro and in silico hybrid approach to unveil triterpenoids from Helicteres hirsuta leaves as potential compounds for inhibiting Nrf2

Cancer is a leading global health concern, with over 20 million new cases and 9.7 million deaths reported in 2022. Chemotherapy remains a widely used treatment, but drug resistance, which affects up to 90% of treatment outcomes, significantly hampers its effectiveness. The transcription factor Nrf2, which is crucial for cellular defense against oxidative stress, plays a dual role in cancer treatment. Although Nrf2 activation can suppress early carcinogenesis, its overexpression in cancer cells contributes to drug resistance, resulting in poor patient outcomes. Thus, inhibiting Nrf2 has emerged as a promising strategy for overcoming cancer drug resistance. Natural compounds such as luteolin and brusatol have shown potential in inhibiting Nrf2, although with limitations. This study isolates and characterizes seven triterpenoids from the n-hexane sub-fraction of Helicteres hirsuta, a plant traditionally used for medicinal purposes, to evaluate their ability to modulate Nrf2 activity in Huh7 cancer and HaCaT normal cells. Additionally, molecular docking and dynamic simulations were utilized to assess the binding potential of these compounds to the PI3K alpha receptor, which regulates downstream signaling pathways, thereby suppressing Nrf2 activity in cancer cells. Our findings provide insights into new strategies seeking triterpenoids as promising structures to reverse chemoresistance by regulating Nrf2. The results also reveal the potential of 3 beta-O-trans-caffeoylbetulinic acid from H. hirsuta leaves as the unprecedented compound inhibiting Nrf2 activity, with an IC50 of 74.5 mu g mL-1 in Huh7 cancer cells.