Development of anti-and pro-apoptotic drugs via regulation of VDAC, a mitochondrial outermembrane protein

电压依赖性阴离子通道(VDAC)是一种存在于线粒体外膜上的通道蛋白。VDAC通过调节线粒体膜通透性，对细胞的存活和死亡起着至关重要的作用。因此，为了开发抗和促凋亡药物，我计划进行两项研究：(1)VDAC调节药物的鉴定；(2)通过调节糖酵解抑制VDAC-己糖激酶相互作用的药物的开发。(1)VDAC调节药物的鉴定-(A)之前，我发现抗凋亡蛋白Bcl2/Bclxl通过直接相互作用调节VDAC。我搜索了结合部位，发现N-末端20个氨基酸(称为BH4区)是至关重要的。将HIV-Tat蛋白衍生的9个氨基酸进行融合，形成细胞通透型。该TAT-BH4多肽关闭了线粒体VDAC通道，并阻止了培养细胞的各种形式的凋亡。此外，当将这种多肽注射到小鼠体内时，辐射诱导的结肠炎、缺血性心脏病和暴发性肝炎都得到了显著改善。(B)为了确定其他VDAC调节剂，我利用分离的线粒体筛选了各种抗生素和小分子。从抗生素文库中，我发现了几个通过调节VDAC通道而取消抗凋亡作用的分子，从小分子筛选中，我发现了抗凋亡试剂。现在，我在体内测试了这些分子的效果。(2)抑制VDAC-己糖激酶相互作用的药物的开发-尽管我尝试了几次筛选，以确定抑制VDAC-己糖激酶相互作用的药物。但是，我还不能确定。

Voltage-dependent anion channel (VDAC) is a channel protein existed on mitochondrial outer membrane. VDAC plays a crucial role for cell living as well as cell death, via regulating mitochondrial membrane permeability. Therefore, in order to develop the anti-and pro-apoptotic drugs, I planned two studies ; (1)Identification of VDAC-regulating drugs, and (2)Development of drugs that inhibit VDAC-hexokinase interaction, which is crucial for cancer cell viability through regulation of glycolysis.(1)Identification of VDAC-regulating drugs-(A)Previously, I found that anti-apoptotic protein Bcl-2/Bcl-xL regulated VDAC through direct interaction. I searched the binding site and found that N-terminal 20 amino acids (called BH4 region) were crucial. To produce cell-permeable form, 9 amino acids derived from HIV-Tat protein were fused. This Tat-BH4 peptide closed VDAC channel in isolated mitochondria, and prevented various forms of apoptosis in cultured cells. Furthermore, when this peptides were administrated into mice, radiation-induced colitis, ischemic heart disease, and fulminant hepatitis were dramatically improved. (B)To identify other VDAC-regulating agents, I screened the various antibiotics and small molecules using isolated mitochondria. From antibiotics library, I found several molecules which cancelled anti-apoptotic Bcl-2 function via regulating VDAC channel, from small molecules screening, I found anti-apoptotic reagent. Now, I tested the effect of these molecules in vivo.(2)Development of drugs that inhibit VDAC-hexokinase interaction-Although I tried several screening to identify the drugs that inhibit VDAC-hexokinase interaction. But, I could not identified yet.