ML-162 is a small molecule that inhibits Glutathione peroxidase 4 (GPX4), with an IC50 value of 1.42µM[1]. ML-162 potently suppresses thioredoxin reductase 1 (TXNRD1) with an IC50 value of 19.5µM[2]. ML-162 has been extensively used as a model compound to explore the covalent binding of ML-162 to the active and catalytic sites of GPX4 and to develop related derivatives[3].
In vitro, ML-162 treatment for 48 hours significantly inhibited the proliferation of HT1080 cells with an IC50 value of 0.6±0.09μM[4]. Treatment of HK-2 cells with 1μM ML-162 for 24 hours significantly inhibited cell viability and induced ferroptosis[5]. Treatment with 10μM ML-162 for 24h significantly caused the cell death of Pfa1 cells and blocked the inhibition of ferroptosis activity of 5mM N-acetyl-l-cysteine (NAC) [6].
In vivo, ML-162 treatment via intravenous injection at a dose of 20mg/kg/day for 19 days significantly inhibited tumor growth in a xenografted BALB/c nude mice model bearing the HT1080 tumor, and reduced GPX4 and Bcl-2 levels in tumor tissues[4]. Combined treatment with 1mg/kg of ML-162 and 10mg/kg of C7 via intraperitoneal injection twice every three days for 21 days significantly inhibited tumor growth in the mouse of HepG2 xenograft model without affecting the body weight of the mice[7]. Daily intraperitoneal injection of ML-162 at a dose of 40mg/kg/day for 2 weeks significantly induced tumor ferroptosis in a mouse model of breast cancer and resulted in increased levels of PTGS2, MDA, and 4-HNE in tumor tissues[8].
References:
[1] Kunishige R, Noguchi Y, Okamoto N, et al. Protein covariation networks for elucidating ferroptosis inducer mechanisms and potential synergistic drug targets[J]. Communications Biology, 2025, 8(1): 480.
[2] Cheff D M, Huang C, Scholzen K C, et al. The ferroptosis inducing compounds RSL3 and ML162 are not direct inhibitors of GPX4 but of TXNRD1[J]. Redox Biology, 2023, 62: 102703.
[3] Moosmayer D, Hilpmann A, Hoffmann J, et al. Crystal structures of the selenoprotein glutathione peroxidase 4 in its apo form and in complex with the covalently bound inhibitor ML162[J]. Biological Crystallography, 2021, 77(2): 237-248.
[4] Ma F, Li Y, Cai M, et al. ML162 derivatives incorporating a naphthoquinone unit as ferroptosis/apoptosis inducers: design, synthesis, anti-cancer activity, and drug-resistance reversal evaluation[J]. European Journal of Medicinal Chemistry, 2024, 270: 116387.
[5] Homma T, Tada C, Yamauchi M, et al. Identification of a novel tetrahydroxynaphthalene derivative by chemical screening with ferroptosis inhibitory activity and promising therapeutic potential[J]. Free Radical Research, 2025, 59(4): 321-331.
[6] Zheng J, Zhang W, Ito J, et al. N-acetyl-l-cysteine averts ferroptosis by fostering glutathione peroxidase 4[J]. Cell Chemical Biology, 2025, 32(5): 767-775. e5.
[7] Zhu J, Tan Q, Fan S, et al. PROTAC degraders of FSP1 act as potent GPX4 sensitizers to induce ferroptosis for hepatoma treatment[J]. Chinese Chemical Letters, 2025: 111285.
[8] Yang F, Xiao Y, Ding J H, et al. Ferroptosis heterogeneity in triple-negative breast cancer reveals an innovative immunotherapy combination strategy[J]. Cell metabolism, 2023, 35(1): 84-100. e8.