D-peptide activators of p53 as anticancer therapeutics

描述（由申请人提供）：拟议的多学科研究结合了当代合成肽/蛋白质化学，结构生物学，癌症生物学以及各种生物化学和生物物理技术，展示了一种强大的，创新的和综合的方法，以填补基于肽的抗癌药物发现的重要空白。如果获得资助，该项目将为现有的抗癌武器库增加新的武器，并广泛影响许多其他人类疾病的靶向分子治疗的肽疗法的发展。背景资料：p53作为转录因子被激活以诱导对细胞应激的强大的生长抑制和凋亡反应，但在正常细胞中由其负调节因子MDM 2和MDMX严格控制。p53通路的受损是几乎所有人类肿瘤的标志，其中TP 53基因突变或p53蛋白被MDM 2和MDMX功能性失活。MDM 2和MDMX在许多肿瘤中的过度表达或扩增与p53的正常（野生型）状态相关，有助于稳健的p53抑制和降解。最近的研究已经验证了抑制p53-MDM 2/MDMX相互作用作为癌症治疗的新的治疗范例。本实验室已开发出一系列高亲和力的MDM 2和MDMX D肽拮抗剂（DPMI-1、DPMI-2、DPMI-3、DPMI-4、DPMI-5），d）当通过脂质体载体媒介物递送时，能够通过激活p53途径在体外和体内杀死肿瘤细胞。目的：利用 DPMI-d作为母体分子（KD = 220 pM的MDM 2和200 nM的MDMX），我们寻求开发高亲和力，蛋白酶抗性，和细胞穿透的D-肽拮抗剂的MDM 2和MDMX的潜在抗癌治疗。具体目的：（1）设计能够穿过细胞膜以稳定细胞内p53的DPMI-d的侧链交联形式。我们将利用碳氢钉合技术设计一系列侧链交联的DPMI-d类似物，并评价它们的功能和结构特性，螺旋度、MDM 2/MDMX结合和膜透化。(2)检验MDM 2和MDMX的高亲和力、蛋白酶抗性和细胞穿透性D肽拮抗剂通过细胞周期阻滞和/或凋亡诱导重新激活p53通路并杀死肿瘤细胞的假设。我们将使用具有或不具有野生型p53的多种肿瘤细胞系来评估侧链交联形式的DPMI-d的肿瘤杀伤活性。我们的长期目标是开发基于D-肽的p53激活剂作为一类新型的抗癌治疗药物用于临床。

DESCRIPTION (provided by applicant): The proposed multi-disciplinary research combines contemporary synthetic peptide/protein chemistry, structural biology, cancer biology, and various biochemical and biophysical techniques, showcasing a powerful, innovative, and integrated approach to filling an important gap in peptide-based anticancer drug discovery. If funded, this project will lead to the addition of new weapons to the existing anticancer arsenal, and broadly impact the development of peptide therapeutics for targeted molecular therapy of many other human diseases as well. Background: p53 is activated as a transcription factor to induce powerful growth inhibitory and apoptotic responses to cellular stress, but is otherwise tightly controlled in normal cells by its negative regulators MDM2 and MDMX. Impairment of the p53 pathway is a hallmark of almost all human tumors where either the TP53 gene is mutated or the p53 protein is functionally inactivated by MDM2 and MDMX. Over-expression or amplification of MDM2 and MDMX in many tumors correlates with a normal (wild type) status of p53, contributing to robust p53 inhibition and degradation. Recent studies have validated inhibition of the p53-MDM2/MDMX interaction as a new therapeutic paradigm for cancer treatment. Our laboratory has developed a series of high-affinity D- peptide antagonists of MDM2 and MDMX (DPMI-¿, ¿, ?, d) that, when delivered via a liposomal carrier vehicle, are capable of killing tumor cells in vitro and in vivo by activating the p53 pathway. Objective: Using DPMI-d as a parent molecule (KD = 220 pM for MDM2 and 200 nM for MDMX), we seek to develop high-affinity, protease-resistant, and cell-penetrating D-peptide antagonists of MDM2 and MDMX for potential anticancer therapy. Specific Aims: (1) Design side-chain cross- linked forms of DPMI-d capable of traversing the cell membrane to stabilize intracellular p53. We will use the hydrocarbon stapling technique to design a series of side-chain cross-linked analogs of DPMI- d, and evaluate their functional and structural properties with respect to ?-helicity, MDM2/MDMX binding, and membrane permeabilization. (2) Test the hypothesis that high-affinity, protease- resistant, and cell-penetrating D-peptide antagonists of MDM2 and MDMX reactivate the p53 pathway and kill tumor cells through cell cycle arrest and/or apoptosis induction. We will evaluate tumor-killing activity of side-chain cross-linked forms of DPMI-d using a variety of tumor cell lines with or without wild type p53. Our long-term objective is to develop D-peptide-based p53 activators as a novel class of anticancer therapeutics for clinical use.