In vitro screening and structural characterization of inhibitors of the S100B-p53 interaction.
In vitro screening and structural characterization of inhibitors of the S100B-p53 interaction.
复制标题
DOI：
10.2147/ijhts.s8210
复制
发表时间：
2010-07-07
期刊：
影响因子：
--
通讯作者：
Weber DJ
中科院分区：
文献类型：
--
作者：
Wilder PT;Charpentier TH;Liriano MA;Gianni K;Varney KM;Pozharski E;Coop A;Toth EA;Mackerell AD;Weber DJ
S100B 在许多癌症中高度过度表达，包括恶性黑色素瘤。在此类癌症中，S100B 以钙依赖性方式结合野生型 p53，将其隔离并促进其降解，导致 p53 依赖性肿瘤抑制活性丧失。因此，S100B抑制剂可能能够恢复某些癌症中野生型p53的水平并提供有用的治疗策略。在这方面，开发并优化了自动化、灵敏的荧光偏振竞争测定（FPCA），以快速筛选结合 Ca2+ 负载的 S100B 并抑制 S100B 靶复合物形成的先导化合物。通过筛选 2000 种化合物，鉴定出 26 种假定的 S100B 低分子量抑制剂。这些小分子与 S100B 的结合通过核磁共振波谱证实，并且通过与 S100B 复合的几种化合物的 X 射线晶体结构提供了额外的结构信息。值得注意的是，许多已鉴定的抑制剂通过化学修饰蛋白质中的 Cys84 发挥作用。这些结果验证了使用高通量 FPCA 来促进抑制 S100B 的化合物的鉴定。这些先导化合物将成为未来优化研究的主题，最终目标是开发一种具有治疗活性的药物，用于治疗恶性黑色素瘤和/或 S100B 升高的其他癌症。
S100B is highly over-expressed in many cancers, including malignant melanoma. In such cancers, S100B binds wild-type p53 in a calcium-dependent manner, sequestering it, and promoting its degradation, resulting in the loss of p53-dependent tumor suppression activities. Therefore, S100B inhibitors may be able to restore wild-type p53 levels in certain cancers and provide a useful therapeutic strategy. In this regard, an automated and sensitive fluorescence polarization competition assay (FPCA) was developed and optimized to screen rapidly for lead compounds that bind Ca2+-loaded S100B and inhibit S100B target complex formation. A screen of 2000 compounds led to the identification of 26 putative S100B low molecular weight inhibitors. The binding of these small molecules to S100B was confirmed by nuclear magnetic resonance spectroscopy, and additional structural information was provided by x-ray crystal structures of several compounds in complexes with S100B. Notably, many of the identified inhibitors function by chemically modifying Cys84 in protein. These results validate the use of high-throughput FPCA to facilitate the identification of compounds that inhibit S100B. These lead compounds will be the subject of future optimization studies with the ultimate goal of developing a drug with therapeutic activity for the treatment of malignant melanoma and/or other cancers with elevated S100B.