Sequence Specific Inhibition of Protein Translation

项目摘要/摘要 药物发现的一个内在挑战是，每个靶点都需要自己的个性化策略。许多 靶标是小分子无法下药的，由于它们起效快，成本低，是 临床治疗和研究工具的药理学基石。一种可推广的解决方案来抑制 因此，任何特定蛋白质与小分子的功能都是生物医学研究的圣杯。一个 可能的解决方案存在于最近描述的一种小分子中，该小分子阻碍了非常有限的 蛋白质靶标的数量。这类分子结合在核糖体出口隧道中，以变构方式改变 肽基转移酶中心和防止伸长，以一种依赖于序列的方式进行 初生链的身份。尽管这种新的作用机制令人兴奋，但电流的脱靶效应 化合物排除了它们作为治疗药物或作为干净的序列特异性化学探针的使用。在这 我们将直接讨论这些化合物能否被开发成有用的研究工具。在……里面 目标1，我们将探索这项技术特异性的内在限制，通过开发一种优化的 已知化合物和进化的目标肽序列之间的化学遗传学配对。在目标2中，我们 将评估是否可以通过开发共价来提高这些化合物的最大功效 可以针对特定的含有半胱氨酸的新生链条的类似物。在目标3中，我们将询问是否 化合物的结构可以被调整，以改变它们所针对的新生链条的序列特异性。 我们将通过一种创新的动力学目标模板化合成策略来实现这一点，该策略将同时探索 结合袋与出口隧道之间可接近的化学空间，同时还产生新生链条 特定的化学探针。我们预计，这些目标中的任何一个目标的进步都将使这项技术能够 进一步发展成为一种强大的工具：能够停止，然后重新启动，任何 感兴趣的目标。

PROJECT SUMMARY/ABSTRACT An inherent challenge in drug discovery is that every target requires its own individualized strategy. Many targets are “undruggable” by small molecules, which, owing to their rapid onset and low cost, are the pharmacologic cornerstone of both clinical therapies and research tools alike. A generalizable solution to inhibit the function of any given protein with a small molecule is therefore a holy grail of biomedical research. A possible solution exists in a recently described small molecule which stalls the translation of a very limited number of protein targets. This class of molecules binds in the ribosome exit tunnel to allosterically alter the peptidyl transferase center and prevent elongation, doing so in a manner that is dependent on the sequence identity of nascent chain. Though this novel mechanism of action is exciting, the off-target effects of the current compounds precludes their use either as therapeutics or as a clean sequence-specific chemical probe. In this proposal, we will directly address whether these compounds can be developed into useful research tools. In Aim 1, we will probe the intrinsic limits of the specificity of this technology, doing so by developing an optimized chemical genetics pair between the known compounds and an evolved target peptide sequence. In Aim 2, we will evaluate whether the maximum efficacy of these compounds can be improved by developing covalent analogs that can target specific cysteine-containing nascent chains. In Aim 3, we will ask whether the structures of the compounds can be tuned to alter the sequence specificity of the nascent chains they target. We will do this via an innovative kinetic target-templated synthetic strategy, which will simultaneously probe the accessible chemical space of the binding pocket with the exit tunnel, while also producing nascent chain specific chemical probes. We anticipate that advances in any one of these Aims would allow this technology to be further developed into a powerful tool: the ability to stop, and then restart, the protein production of any target of interest.