ML-162

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.

ML-162是一种可抑制谷胱甘肽过氧化物酶4（GPX4）的小分子化合物，IC₅₀值为1.42µM^[1]。ML-162能强效抑制硫氧还蛋白还原酶1（TXNRD1），IC₅₀值为19.5µM^[2]。ML-162已被广泛用作模型化合物，用于探索与GPX4活性和催化位点的共价结合机制，并开发相关衍生物^[3]。

在体外，ML-162处理48小时可显著抑制HT1080细胞增殖，IC₅₀值为0.6±0.09μM^[4]。用1μM的ML-162处理HK-2细胞24小时，能显著抑制细胞活力并诱导铁死亡^[5]。使用10μM的ML-162处理Pfa1细胞24小时，可显著引起细胞死亡，并阻断5mM N-乙酰-L-半胱氨酸（NAC）对铁死亡的抑制作用^[6]。

在体内，每日静脉注射20mg/kg剂量的ML-162，连续19天，能显著抑制携带HT1080肿BALB/c裸鼠模型的肿瘤生长，并降低肿瘤组织中GPX4和Bcl-2的水平^[4]。在HepG2移植瘤小鼠模型中，每三天腹腔注射2次1mg/kg的ML-162与10mg/kg的C7（联合治疗），持续21天，可显著抑制肿瘤生长且不影响小鼠体重^[7]。在乳腺癌小鼠模型中，每日腹腔注射40mg/kg剂量的ML-162，连续2周，能显著诱导肿瘤铁死亡，导致肿瘤组织中PTGS2、MDA和4-HNE水平升高^[8]。