Discovery of incednine as a potent modulator of the anti-apoptotic function of Bcl-xL from microbial origin
Discovery of incednine as a potent modulator of the anti-apoptotic function of Bcl-xL from microbial origin
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DOI：
10.1021/ja710124p
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发表时间：
2008-02-13
影响因子：
15
通讯作者：
Imoto, Masaya
中科院分区：
文献类型：
--
作者：
Futamura, Yushi;Sawa, Ryuichi;Imoto, Masaya
抗凋亡癌蛋白Bcl-2和BcI-xL在许多癌症中过表达，并在癌症的发生、进展和对化疗的抵抗中起关键作用。因此，发现这些蛋白质的功能性抑制剂并改善对这些蛋白质的分子机制的理解将有助于新的抗肿瘤疗法。在这里，使用化学遗传学细胞为基础的筛选，我们已经发现了一种化学和生物学上独特的物质，incednine，作为一种新的功能调节Bcl-2/Bcl-xL从链霉菌的发酵液。ML-693- 90 F3。该化合物通过溶剂萃取和使用离心液-液分配色谱法分离为HCl盐。通过光谱分析、X射线晶体学分析和计算研究阐明了其结构。Incednine的分子式为C42 H63 N3 O 8，由一个新的骨架结构组成，在一个24元大环内酰胺核中的烯醇-醚酰胺，以及两个氨基糖。Bcl-xL-过表达的Ms-1细胞表现出对几种抗肿瘤药物的抗性;然而，仅当细胞用incednine处理时才观察到抗肿瘤药物诱导的细胞死亡。Bcl-xL的过表达通过与Bax形成复合物来抑制过表达Bax的HEK 293 T细胞中的细胞死亡，而Bcl-xL在incednine存在下不能抑制细胞死亡，而不影响Bcl-xL和Bax的异源二聚化。这些发现表明，incednine作为一个有效的调节剂的抗凋亡Bcl-2/Bcl-xL不同于其他已知的Bcl-2抑制剂，并可能提供一个化学探针，以研究Bcl-2/Bcl-xL的潜在机制。
Anti-apoptotic oncoproteins Bcl-2 and BcI-xL are overexpressed in many cancers and play a crucial role in cancer initiation, progression, and resistance to chemotherapy. Therefore, the discovery of a functional inhibitor for these proteins and improved understanding of the molecular mechanisms of these proteins will be an aid to novel anti-tumor therapies. Here, using chemical-genetic cell-based screening, we have discovered a chemically and biologically unique substance, incednine, as a novel functional modulator of Bcl-2/Bcl-xL from the fermentation broth of Streptomyces sp. ML-693-90F3. This compound was isolated as a HCI salt by solvent extraction and using centrifugal liquid-liquid partition chromatography. Its structure was elucidated by spectroscopic analysis, X-ray crystallographic analysis, and computational studies. Incednine has a molecular formula, C42H63N3O8, and consists of a novel skeletal structure, enol-ether amide in a 24-membered macrolactam core, with two aminosugars. Bcl-xL-overexpressing Ms-1 cells displayed resistance to several anti-tumor agents; however, anti-tumor agent-induced cell death was observed only when cells were treated with incednine. Overexpression of BcI-xL inhibited cell death in Bax-overexpressing HEK293T cells by forming a complex with Bax, whereas Bcl-xL failed to inhibit cell death in the presence of incednine without affecting the heterodimerization of Bcl-xL and Bax. These findings suggest that incednine serves as a potent modulator of the anti-apoptotic Bcl-2/Bcl-xL distinct from the other known Bcl-2 inhibitors and could provide a chemical probe to study the underlying mechanisms of Bcl-2/Bcl-xL.