Selective inhibitors of deubiquitylases

描述(申请人提供)：在过去的十年中，泛素研究领域有了爆炸性的增长，大约有530个人类基因被预测为编码参与泛素的结合和去结合的酶。其中95个编码脱泛素酶(DUB)，这些酶已经成为许多正在进行的药物发现努力的目标。所有的DUBS都进行酶促去除目标蛋白中的泛素(Ub)，无论该蛋白是正常的细胞蛋白还是Ub本身。解除纠纷需要 在Ub的C-末端和目标蛋白中赖氨酸残基的-NH2-基团之间形成的异肽键的裂解。除了JAMM-DUBS家族外，所有的DUB都属于肽酶C19家族，并且在活性部位有一个激活的半胱氨酸残基。因此，大多数筛选程序对表现出低选择性和低效力的环状活性化合物的命中率很高。最有效的DUB抑制剂通常是通过在Ub的C末端掺入Cys反应部分而产生的，例如Ub-醛、Ub-乙烯基砜、Ub-乙烯基甲酯。根据它们的性质，即依赖于Ub与酶的结合，这些抑制剂在不同的DUB之间表现出很少或没有选择性。Ub研究领域和DUB作为药物发现努力的目标的确认，将因特定DUB的选择性、高效抑制剂的可获得性而显著推进。在这一应用中，我们建议开发此类抑制剂的面板。为了达到高效力，我们将通过使用β-内酰胺支架来利用活性部位半胱氨酸的反应性。活性中心Cys对内酰胺环的攻击导致稳定的四面体中间体的生成。选择性是通过用于装饰环的取代基来获得的。这种方法已经被其他人用来生产许多酶的高选择性抑制剂，例如凝血酶激活因子X(FXA)和激活因子XI(FXIa)、白细胞弹性蛋白酶、类胰蛋白酶和组织蛋白酶。应该注意的是，组织蛋白是半胱氨酸蛋白酶，表明内酰胺类抑制剂将对这一机械类别起作用。虽然我们在这个项目中的主要目标是开发一系列新的研究工具，但我们生产的化合物可以用作药物开发的先导化合物。最后，我们围绕这些化合物开发的SAR可以用于指导围绕其他化合物的药物化学努力。

DESCRIPTION (provided by applicant): In the last decade there has been an explosive growth in the field of ubiquitin research, with approximately 530 human genes predicted to encode enzymes involved in the conjugation and deconjugation of ubiquitin. Of these 95 encode deubiquitylases (DUBs) and these enzymes have become the target of many on-going drug discovery efforts. All DUBs carry-out the enzymatic removal of ubiquitin (Ub) from target proteins, whether that protein is a normal cellular protein or is Ub itself. Deconjugation requires the cleavage of an isopeptide bond formed between the C-terminus of Ub and the ¿-NH2-group of a Lys residue in the target protein. With the exception of the JAMM-family of DUBs, all DUBs fall into the peptidase C19 family and have an activated cysteine residue in the active site. As a consequence, most screening programs have a high hit rate of cys-reactive compounds that show low selectivity and potency. The most potent DUB inhibitors are generally produced by incorporation of a cys-reactivity moiety at the C-terminus of Ub, e.g. Ub-aldehyde, Ub-vinylsulfone, Ub-vinylmethylester. By their very nature, i.e. relying on the binding of Ub to the enzyme, such inhibitors exhibit little or no selectivity between different DUBs. The field of Ub research and the validation of DUBs as targets for drug discovery efforts, would be dramatically advanced by the availability of selective, high potency inhibitors of specific DUBs. In this application, we propose to develop panels of such inhibitors. In order to achieve high potency, we will take advantage of the reactivity of the active site cysteine through the use of a ¿-lactam scaffold. Attack on the lactam ring by the active site Cys leads to the generation of a stable tetrahedral intermediate. Selectivity is obtained by the substituents used to decorate the ring. This approach has been used by others to produce highly selective inhibitors of a number of enzymes, e.g. the blood coagulation enzymes activated factor X (FXa) and activated factor XI (FXIa), leukocyte elastase, tryptase, and cathepsins. It should be noted that the cathepsins are cysteine proteases, demonstrating that lactam-based inhibitors will function against this mechanistic class. Although our primary goal in this project is to generate a new series of research tools, the compounds that we produce could be used as lead compounds for drug development. Finally, the SAR that we develop around these compounds can be used to guide Medicinal Chemistry efforts around other compounds.