Ferroptosis-IN-22 is a selective inhibitor of ferroptosis, which functions by targeting NCOA4 and disrupting its interaction with ferritin, exhibiting an EC50 of 520 nM and a Kd value of 0.78 μM. It effectively inhibits ferroptosis induced by various agents (RSL3, Erastin, ML210, FIN56) but does not inhibit cell necrosis induced by H2O2 or apoptosis induced by STS. Additionally, Ferroptosis-IN-22 significantly ameliorates Concanavalin A-induced acute liver injury and can be used in the study of ferroptosis-related diseases.

Ferroptosis-IN-22 (Compound 4k) exhibits anti-ferroptosis activity with EC₅₀ values of 0.515 μM (RSL3) and 2.776 μM (Erastin) in HT-1080 cells, 0.4061 μM (RSL3) and 1.308 μM (Erastin) in 786-O cells, and 0.093 μM (RSL3) and 1.013 μM (Erastin) in MDA-MB-231 cells, also inhibiting ferroptosis induced by ML210 and FIN56. At concentrations of 4–8 μM, it significantly reduces lipid peroxidation in HT-1080 and 786-O cells induced by RSL3 or Erastin, without having direct antioxidant activity. Ferroptosis-IN-22 (4 μM, 6 hours) binds directly to the key receptor protein NCOA4 in ferritinophagy, disrupting the interaction between NCOA4 and ferritin (FTH1), thus inhibiting the process in HT-1080 cells independently of direct iron chelation. It also shows low cardiac toxicity and a very low risk of CYP-mediated drug-drug interactions (DDI).

Ferroptosis-IN-22 (Compound 4k) administered intraperitoneally at doses of 5-15 mg/kg twice before ConA injection can alleviate acute liver injury induced by ferroptosis, triggered by concanavalin A (ConA).