TARGET-GUIDED DEVELOPMENT OF SPECIFIC AKT INHIBITORS

描述（由申请人提供）：我们计划使用原位点击化学法生产高度特异性和有效的Akt激酶抑制剂，这是一种采用靶蛋白从简单前体组装抑制剂的新技术。Akt是一种丝氨酸苏氨酸蛋白激酶，在细胞生长和复制的控制中起核心作用。它在大约50%的非小细胞肺癌中显示出功能的增加。乳腺癌、前列腺癌和胰腺癌也显示出Akt的参与。因此，Akt抑制剂的潜在临床适应症广泛且众多。原位点击化学允许产生对Akt具有无与伦比的特异性和效力的双配体抑制剂。该方法的原理是设计两个配体靶向酶上的两个相邻位点。配体配备有互补的反应基团，并且在两个配体同时结合到它们的靶标时，这些反应基团形成共价连接，导致形成双配体，其与蛋白质的结合口袋进行多重相互作用。具体目标是：（1）设计并产生靶向ATP结合位点并携带用于原位点击化学的额外官能团的化合物文库;在体外筛选这些文库以获得将用作锚分子的ATP竞争性Akt抑制剂。(2)用与Akt结合的锚分子进行原位点击化学并产生二价Akt配体。(3)确定二价Akt抑制剂的效力和特异性，进行基于细胞的测定以干扰Akt诱导的致癌转化。(4)通过原位点击化学研究结构-活性关系，并优化抑制剂的效力，选择性和细胞活性

DESCRIPTION (provided by applicant): We plan to produce highly specific and potent inhibitors of the Akt kinase using in situ click chemistry, a new technique that employs the target protein for assembling inhibitors from simple precursors. Akt is a serinethreonine protein kinase that plays a central role in the control of cell growth and replication. It shows a gain of function in about 50 percent of the non-small cell lung cancers. Cancer of the breast, prostate and pancreas also show involvement of Akt. The potential clinical indications for Akt inhibitors are, therefore, wide and numerous. In situ click chemistry allows the generation of biligand inhibitors of unsurpassed specificity and potency for Akt. The principle of this method is to design two ligands targeted to two neighboring sites on the enzyme. The ligands are equipped with complementary reactive groups, and upon simultaneous binding of the two ligands to their targets, these reactive groups form a covalent connection, causing the formation of a biligand, which engages in multiple interactions with the protein's binding pockets. The specific aims are: (1) Design and produce libraries of compounds that are targeted at the ATP-binding site and that carry additional functional groups for in situ click chemistry; screen these libraries in vitro for ATP-competitive Akt inhibitors that will be used as anchor molecules. (2) Perform in situ click chemistry with anchor molecules bound to Akt and generate bivalent Akt ligands. (3) Determine potency and specificity of bivalent Akt inhibitors, perform cell-based assays for interference with Akt-induced oncogenic transformation. (4) Investigate the structure-activity relationship through in situ click chemistry and optimize inhibitors for potency, selectivity and cell-based activity