通过分子剂调节14-3-3σ/C-RAF"自动"抑制复合物
Markella Konstantinidou1, Holly R Vickery1, Marloes A M Pennings2
1Department of Pharmaceutical Chemistry and Small Molecule Discovery Center (SMDC), University of California, San Francisco 94143, United States.
bioRxiv : the preprint server for biology
|August 6, 2025
概括
研究人员开发了新的分子粘剂来稳定C-RAF/14-3-3蛋白相互作用,这是MAPK通路障碍的关键目标. 这种方法通过调节蛋白质-蛋白质相互作用,为瘤学和RAS病提供了新的治疗策略.
科学领域:
- 化学生物学 化学生物学
- 药物发现 药物发现 药物发现
- 分子生物学分子生物学
背景情况:
- MAPK信号通路,特别是C-RAF,在瘤学和RAS病症中至关重要,但直接激酶抑制存在局限性.
- 调节蛋白质-蛋白质相互作用 (PPI),比如C-RAF和14-3-3之间的调节,是一个尚未探索的治疗途径.
- C-RAF的14-3-3结合胺259调节其激活,使得这种PPI成为一个关键的监管点.
研究的目的:
- 设计和验证稳定原生C-RAF/14-3-3蛋白质-蛋白质相互作用的分子.
- 探索通过PPI稳定来准本质上混乱的地区的潜力.
- 为研究MAPK路径和开发新疗法提供化学探针.
主要方法:
- 碎片合并方法用于设计针对14-3-3/C-RAF复合物的分子接架.
- 生物物理试验量化复杂稳定性,并评估RAF异型和14-3-3客户端之间的选择性.
- 三元复合物的晶体结构,以证实基于的作用机制.
- 基于细胞的测试以验证在目标上的参与和下游信号效应.
主要成果:
- 合成的分子粘剂使14-3-3/C-RAF复合物稳定到300倍.
- 通过晶体结构证实了基于的机制,并对目标PPI表现出高的选择性.
- 细胞测定显示了目标参与,增加了C-RAF pS259酸化,减少了RAF二分化,减少了ERK酸化.
结论:
- 分子剂有效地稳定了C-RAF/14-3-3相互作用,提供了一种超出直接激酶抑制的新策略.
- 这种方法可以准以前无法访问的蛋白质接口,并提供有价值的化学探针.
- 开发的分子合剂作为在瘤学和RASopathies药物发现的起点.
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