Kinases as Therapeutic Targets for Endometriosis

摘要 人类基因组编码538个蛋白激酶，其中大部分在功能上是重要的，并参与 许多疾病的发病机制。这一新的R01是一项基于发现的拨款，专注于分子 子宫内膜异位症中上调的新蛋白激酶的验证。解决新的治疗方案 子宫内膜异位症，我们将使用siRNA敲除来验证与子宫内膜异位症相关的激酶的子集 策略和可用的小分子抑制剂，并随后使用DNA编码的化学技术 (DEC-Tec)，以确定新的治疗分子，以便在体外和体内进一步进行化学验证。我们 确定了数十种在卵巢疾病患者皮损中统计上上调的蛋白激酶 子宫内膜异位症、腹膜子宫内膜异位症和/或深度浸润性子宫内膜异位症与对照组和患者的比较 子宫内膜。R01的假设是，抑制一个或多个这些新的激酶靶点将是 对治疗子宫内膜异位症有效。在这项提案中，我们概述了一种全面的方法来 原代子宫内膜异位间质细胞和器质性培养中的功能特征和靶向蛋白激酶 源自女性。同时，我们将利用最先进的DEC-Tec选择方法，可在 位于贝勒医学院(BCM)的药物发现中心(CDD)，以确定可 然后在没有DNA条形码的情况下重新合成，并由我们的团队进行化学优化。我们目前有 超过60亿个DNA编码分子的筛选集合。首要目标是找出有效的 和选择性的激酶抑制剂，可以为解决未满足的临床问题的新药提供基础 子宫内膜异位症患者的需求。其具体目标是：1)对过表达的激酶进行靶向验证； 2)使用DEC-Tec来鉴定新的有效的激酶的小分子抑制剂并优化选择性， 高亲和力激酶结合剂的活性和稳定性；以及3)评估优化的高亲和力激酶抑制剂 体外和体内。寻找治疗子宫内膜异位症的新的有效疗法需要多方面的方法， 需要加大努力来确定和筛选药物发现的新靶点。我们的方法，即 将用于靶标识别的基因表达数据集与新型DEC-Tec药物发现平台相结合，Will 通过完善的药物发现管道快速推进关键的激酶靶点。我们的多学科 研究小组正在寻求发现新的化学实体来解决子宫内膜异位症的非激素轴，如 在间歇期单独治疗或作为内分泌抑制药的序贯治疗。我们的DEC-Tec Platform将帮助我们找到第一种未探索的激酶的特定抑制剂(一种主要的可用药类别 蛋白质)在子宫内膜异位症中升高，以剖析这些关键的生理途径和病理 研究子宫内膜异位症的潜在机制，并提供新的针对疾病的治疗方案。

ABSTRACT The human genome encodes 538 protein kinases, most of which are functionally important and are involved in the pathogenesis of many diseases. This new R01 is a discovery-based grant focused on the molecular validation of novel kinases that are upregulated in endometriosis. To address new therapeutic options for endometriosis, we will validate a subset of endometriosis-implicated kinases using siRNA knockdown strategies and available small molecule inhibitors and subsequently use DNA-Encoded Chemistry Technology (DEC-Tec) to identify novel therapeutic molecules for further chemical validation in vitro and in vivo. We identified dozens of kinases that are statistically upregulated in the lesions of patients with ovarian endometriosis, peritoneal endometriosis, and/or deep infiltrating endometriosis compared to control and patient endometrium. The hypothesis for this R01 is that inhibition of one or more of these novel kinase targets will be effective in the treatment of endometriosis. In this proposal, we outline a comprehensive approach to functionally characterize and target kinases in primary endometriotic stromal cells and organoid cultures derived from women. In parallel, we will utilize state-of-the-art DEC-Tec selection methods available in the Center for Drug Discovery (CDD) at Baylor College of Medicine (BCM) to identify novel kinase inhibitors that will then be re-synthesized without the DNA barcode and chemically optimized by our team. We currently have a screening collection of over 6 billion DNA-encoded molecules. The overarching objective is to identify potent and selective kinase inhibitors that can provide a foundation for new medicines to address the unmet clinical needs of endometriosis patients. The Specific Aims are: 1) Perform target validation of overexpressed kinases; 2) Use DEC-Tec to identify novel small-molecule inhibitors of validated kinases and optimize the selectivity, activity, and stability of high affinity kinase binders; and 3) Evaluate optimized high affinity kinase inhibitors in vitro and in vivo. Identifying new efficacious therapies for endometriosis requires a multifaceted approach that necessitates increased efforts to identify and screen novel targets for drug discovery. Our approach, which combines gene expression datasets for target identification with a novel DEC-Tec drug discovery platform, will rapidly advance key kinase targets through a well-established drug discovery pipeline. Our multidisciplinary team is seeking to discover new chemical entities that address the non-hormonal axes of endometriosis as stand-alone therapy or as sequential therapy during hiatus from endocrine-suppressing agents. Our DEC-Tec platform will help us to find the first specific inhibitors of unexplored kinases (a major druggable class of proteins) that are elevated in endometriosis to dissect these key physiological pathways and pathological mechanisms underlying endometriosis and provide new disease-specific treatment options.