DHODH-IN-33 is a selective DHODH inhibitor that exhibits potent activity against A549 cells (IC50 = 5.22 μM) and 5637 cells (IC50 = 3.03 μM). It induces autophagy-dependent ferroptosis, characterized by mitochondrial dysfunction, lipid peroxidation, and accumulation of ROS, without significant toxicity in vivo. The anticancer effect of DHODH-IN-33 is achieved through promoting autophagy-dependent degradation of dihydroorotate dehydrogenase. This compound can be used in studies related to non-small cell lung cancer and bladder cancer.

DHODH-IN-33 (compound 3af) demonstrates potent activity against A549 cells (IC 50 = 5.22 μM) and 5637 cells (IC 50 = 3.03 μM) over 48 hours. However, at concentrations of 5 and 10 μM for 48 hours, it induces A549 cell death without involving apoptotic pathways. When applied at 5 and 10 μM for 24 hours, DHODH-IN-33 links the induction of ferroptosis to the activation of autophagy in A549 cells, primarily through the coordinated degradation of redox-stabilizing enzymes. Moreover, at these concentrations over 48 hours, it promotes autophagy-dependent ferroptosis by initiating autophagy, inhibiting DHODH, and triggering lipid peroxidation, serving as the primary death mechanism in A549 cells. These processes act as upstream switches to drive downstream lipid peroxidation execution events.

DHODH-IN-33 (compound 3af) (10 and 20 mg/kg, intraperitoneal injection every 48 hours for 14 days) exhibits antitumor activity in a subcutaneous A549 xenograft mouse model primarily through an autophagy-dependent ferroptosis pathway, with ferroptosis as the main mechanism and autophagy activation as a crucial prerequisite. It also maintains good tolerability in BALB/c nude mice.