Discovery of Small Molecule Bak Activator for Lung Cancer Therapy.
Discovery of Small Molecule Bak Activator for Lung Cancer Therapy.
复制标题
发现用于肺癌疗法的小分子BAK激活剂。
作者：
Park D;Anisuzzaman ASM;Magis AT;Chen G;Xie M;Zhang G;Behera M;Sica GL;Ramalingam SS;Owonikoko TK;Deng X
基本原理：巴克是一个主要的促凋亡Bcl 2家族成员和凋亡细胞死亡所需的分子。在小细胞肺癌（SCLC）和非小细胞肺癌（NSCLC）细胞系中均观察到高水平的内源性巴克。巴克蛋白在非小细胞肺癌中的表达与患者的预后密切相关，提示巴克蛋白是一个有吸引力的肺癌治疗靶点。BH 3结构域作为死亡结构域起作用，并且是巴克启动凋亡性细胞死亡所必需的。因此，BH 3结构域是发现巴克激动剂的有吸引力的靶标。研究方法：选择巴克的BH 3死亡结构域结合口袋（aa 75 -88）作为对接位点，用于使用UCSF DOCK 6.1程序套件和NCI化学文库（300，000个小分子）数据库筛选小分子巴克活化剂。从NCI获得确定对BH 3结构域具有最高亲和力的前500种化合物，并测试细胞毒性以进行进一步筛选。我们鉴定了小分子巴克激活剂BKA-073作为先导化合物。通过等温滴定量热法（ITC）分析BKA-073与巴克蛋白的结合亲和力。通过与双（马来酰亚胺基）己烷（BMH）交联分析BKA-073介导的经由寡聚化的巴克活化。通过动态BH 3谱（DBP）和凋亡细胞死亡试验评价BKA-073对肺癌细胞的体外敏感性。在动物模型中评估BKA-073单独或与放射疗法或Bcl 2抑制剂组合的效力。结果：BKA-073与巴克在BH 3结构域有高亲和力和选择性结合。BKA-073/巴克结合促进巴克寡聚化和激活其促凋亡功能的线粒体引发。BKA-073在小细胞肺癌（SCLC）和NSCLC异种移植物、患者源性异种移植物和突变型KRAS驱动癌症的基因工程小鼠模型中有效抑制肿瘤生长，无显著正常组织毒性。巴克在放射抗性肺癌细胞中积累，BKA-073逆转放射抗性。BKA-073与Bcl-2抑制剂venetoclax的组合在体内表现出对肺癌的强协同作用。结论：小分子巴克激活剂的开发为肺癌的治疗提供了新的思路。
Rationale: Bak is a major proapoptotic Bcl2 family member and a required molecule for apoptotic cell death. High levels of endogenous Bak were observed in both small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) cell lines. Increased Bak expression was correlated with poor prognosis of NSCLC patients, suggesting that Bak protein is an attractive target for lung cancer therapy. The BH3 domain functions as death domain and is required for Bak to initiate apoptotic cell death. Thus, the BH3 domain is attractive target for discovery of Bak agonist. Methods: The BH3 death domain binding pocket (aa75-88) of Bak was chosen as a docking site for screening of small molecule Bak activators using the UCSF DOCK 6.1 program suite and the NCI chemical library (300,000 small molecules) database. The top 500 compounds determined to have the highest affinity for the BH3 domain were obtained from the NCI and tested for cytotoxicity for further screening. We identified a small molecule Bak activator BKA-073 as the lead compound. The binding affinity of BKA-073 with Bak protein was analyzed by isothermal titration calorimetry (ITC) assay. BKA-073-mediated Bak activation via oligomerization was analyzed by a cross-linking with Bis (maleimido) hexane (BMH). Sensitivity of BKA-073 to lung cancer cells in vitro was evaluated by dynamic BH3 profiling (DBP) and apoptotic cell death assay. The potency of BKA-073 alone or in combination with radiotherapy or Bcl2 inhibitor was evaluated in animal models. Results: We found that BKA-073 binds Bak at BH3 domain with high affinity and selectivity. BKA-073/Bak binding promotes Bak oligomerization and mitochondrial priming that activates its proapoptotic function. BKA-073 potently suppresses tumor growth without significant normal tissue toxicity in small cell lung cancer (SCLC) and NSCLC xenografts, patient-derived xenografts, and genetically engineered mouse models of mutant KRAS-driven cancer. Bak accumulates in radioresistant lung cancer cells and BKA-073 reverses radioresistance. Combination of BKA-073 with Bcl-2 inhibitor venetoclax exhibits strong synergy against lung cancer in vivo. Conclusions: Development of small molecule Bak activator may provide a new class of anticancer agents to treat lung cancer.
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影响因子：
20.4
作者：
Coate, Linda E.;Massey, Christine;Shepherd, Frances A.
通讯作者：
Shepherd, Frances A.
影响因子：
2.1
作者：
Guerra, Veronica A.;DiNardo, Courtney;Konopleva, Marina
通讯作者：
Konopleva, Marina
影响因子：
15.9
作者：
Chen, Guo;Magis, Andrew T.;Deng, Xingming
通讯作者：
Deng, Xingming
DOI：
10.1007/978-1-4939-8861-7_4
发表时间：
2019-01-01
期刊：
BCL-2 FAMILY PROTEINS
影响因子：
--
作者：
Fraser, Cameron;Ryan, Jeremy;Sarosiek, Kristopher
通讯作者：
Sarosiek, Kristopher
影响因子：
64.8
作者：
Ji, Hongbin;Ramsey, Matthew R.;Wong, Kwok-Kin
通讯作者：
Wong, Kwok-Kin