Hepatocellular carcinoma (HCC) is one of most common and deadliest malignancies. Celastrol (Cel), a natural product derived from the Tripterygium wilfordii plant, has been extensively researched for its potential effectiveness in fighting cancer. However, its clinical application has been hindered by the unclear mechanism of action. Here, we used chemical proteomics to identify the direct targets of Cel and enhanced its targetability and anti-tumor capacity by developing a Cel-based liposomes in HCC. We demonstrated that Cel selectively targets the voltage-dependent anion channel 2 (VDAC2). Cel directly binds to the cysteine residues of VDAC2, and induces cytochrome C release via dysregulating VDAC2-mediated mitochondrial permeability transition pore (mPTP) function. We further found that Cel induces ROS-mediated ferroptosis and apoptosis in HCC cells. Moreover, coencapsulation of Cel into alkyl glucoside-modified liposomes (AGCL) improved its antitumor efficacy and minimized its side effects. AGCL has been shown to effectively suppress the proliferation of tumor cells. In a xenograft nude mice experiment, AGCL significantly inhibited tumor growth and promoted apoptosis. Our findings reveal that Cel directly targets VDAC2 to induce mitochondria-dependent cell death, while the Cel liposomes enhance its targetability and reduces side effects. Overall, Cel shows promise as a therapeutic agent for HCC.

肝细胞癌（HCC）是最常见和最致命的恶性肿瘤之一。雷公藤红素 (Cel) 是一种从雷公藤植物中提取的天然产物，因其抗癌的潜在功效而被广泛研究。然而，其作用机制尚不明确，阻碍了其临床应用。在这里，我们利用化学蛋白质组学来识别 Cel 的直接靶点，并通过开发基于 Cel 的肝癌脂质体来增强其靶向性和抗肿瘤能力。我们证明 Cel 选择性地靶向电压依赖性阴离子通道 2 (VDAC2)。 Cel 直接与 VDAC2 的半胱氨酸残基结合，并通过失调 VDAC2 介导的线粒体通透性转换孔 (mPTP) 功能诱导细胞色素 C 释放。我们进一步发现 Cel 诱导 HCC 细胞中 ROS 介导的铁死亡和细胞凋亡。此外，将Cel共封装到烷基糖苷修饰脂质体（AGCL）中可以提高其抗肿瘤功效并最大限度地减少其副作用。 AGCL已被证明可以有效抑制肿瘤细胞的增殖。在异种移植裸鼠实验中，AGCL显着抑制肿瘤生长并促进细胞凋亡。我们的研究结果表明，Cel 直接靶向 VDAC2 以诱导线粒体依赖性细胞死亡，而 Cel 脂质体增强其靶向性并减少副作用。总体而言，Cel 显示出作为 HCC 治疗剂的前景。