These findings demonstrate that ML210 potentiates Tempol's pro-oxidant pressure by targeting GPX4, selectively amplifying H2O2 accumulation and ER stress engagement without collapsing global redox balance. This study provides mechanistic rationale for redox-proteostasis co-targeting in gastric and colon cancers and establishes a foundation for in vivo validation.

In the CT26 mouse model, ML210-loaded prodrug nanoassemblies demonstrated superior antitumor effects. The strategy-using prodrug as "carriers" for ferroptosis inducers-offers a promising approach for synergistic antitumor therapy.

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SIGNIFICANCE STATEMENT: The implications of this study are that a small molecule of furazan nitrogen oxide of nitric oxide donor coupled with aurovertin B hybrids, 4D, inhibited GPX4 through covalently binding to GPX4 to induce ferroptosis in acute T lymphocytic leukemia cells. This study identified a promising drug candidate for the development of an anti-T-cell acute lymphoblastic leukemia agent in the future.

In vivo pharmacodynamic evaluation demonstrated that Y19 exhibited superior antitumor efficacy compared to RSL3 at an equivalent dose, achieving a tumor growth inhibition (TGI) rate of 72.53 % at 10 mg/kg in the MDA-MB-231 xenograft model, with no significant toxicity observed. This provides structurally diverse tool compounds to reveal the basic biology of GPX4 as well as to explore potential efficacy with the treatment of cancer by inducing ferroptosis.

Notably, ferroptosis inhibitors (ferrostatin-1 and deferoxamine mesylate) could reverse the increase of lipid ROS induced by W25, suggesting that W25 could selectively trigger ferroptosis in HCT-116 cells...Furthermore, MG132 (a proteasome inhibitor) reversed the inhibition of GPX4 protein expression by W25. Correspondingly, the immunoprecipitation assay also showed that W25 promoted the ubiquitination level of GPX4, revealing that W25 induced ubiquitination-dependent proteasomal degradation of GPX4. These results suggest that W25 is a novel ferroptosis inducer against colorectal cancer through the inhibition of GPX4.

Our results indicate that TXNIP is a key player in ferroptotic pathway, as its deletion conferred resistance to classic ferroptosis-inducing agents (erastin, RSL3, and ML210), while TXNIP overexpression increased their susceptibility to ferroptosis. Our findings suggest that TXNIP acts as a positive regulator of ferroptosis by modulating autophagy and iron availability. Targeting TXNIP might hold promise in developing drugs for diseases involving the ferroptotic pathway.

ML210-ansaFc exhibited robust tumor growth suppression in 3D spheroid models, coupled with favorable drug-like properties, highlighting its potential as a therapeutic agent for intractable cancers. This work could pave the way for the development of metallocene-based chemotypes with diverse spatial configurations for the treatment of multiple diseases.

Treatment with Ferrostatin-1 partially rescued the cell viability reduced by DHC. Collectively, these findings suggest that DHC may be a potential agent for inducing ferroptosis in prostate cancer cells by decreasing GPX4 expression.

The combined application of GPx4 and Mitogen-activated protein kinase kinase (MEK) inhibitors, namely ML210 and trametinib, respectively, reduced lethality and tumor-like phenotypes in these flies. Notably, this combination treatment synergistically suppressed the proliferation of human PDAC cells and their corresponding xenografts in mice by inducing ROS accumulation, which triggered ferroptosis. These results suggest that inducing ferroptosis could represent a promising therapeutic strategy for PDAC.

GPX4 suppression, alone or with current TNBC therapies, impacts outcomes in preclinical TNBC models with or without obesity and offers a new, plausible mechanistic target for TNBC treatment.

The oxidization of the cell membrane lipids may suppress EMT, including cell migration. Since EMT is a major malignant phenotype of PDAC, our findings may lead to the advancement of PDAC therapy, especially in the prevention of postoperative recurrence.