Dual inhibition of MDM2 and XIAP as a therapeutic strategy in cancer

MDM2 和 XIAP 双重抑制作为癌症治疗策略

• 批准号: 10224705
• 负责人: WEI LI
• 金额: $ 54.15万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-28 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10224705
• 关键词: Antibodies; Apoptosis; Apoptotic; BIRC4 gene; Back; Binding; Biological; Biological Assay; Biotechnology; C-terminal; CRISPR/Cas technology; Calorimetry; Cell Line; Cell Survival; Cessation of life; Clinical; Complex; Computer Assisted; Crystallization; Disease Progression; Down-Regulation; Drug Design; Drug resistance; Evaluation; Fluorescence Polarization; Genes; Goals; Human; Immune; Induction of Apoptosis; Internal Ribosome Entry Site; Knock-out; Leukemic Cell; MDM2 gene; Malignant Neoplasms; Measurement; Messenger RNA; Molecular; Molecular Biology; Mus; Neuroblastoma; Normal Cell; Oncoproteins; Pharmaceutical Chemistry; Play; Property; Proteins; Publishing; Resistance development; Roentgen Rays; Role; SCID Mice; Structure; Structure-Activity Relationship; Surface Plasmon Resonance; TP53 gene; Testing; Therapeutic; Tissues; Titrations; Toxic effect; Transfection; Translations; Treatment outcome; analog; anti-cancer; anticancer activity; anticancer treatment; base; cancer cell; cancer drug resistance; cancer therapy; cancer type; design; efficacy study; genome editing; high throughput screening; in vivo; inhibitor/antagonist; leukemia; molecular modeling; mouse model; neoplastic cell; neuroblastoma cell; novel; novel therapeutics; overexpression; pharmacokinetics and pharmacodynamics; preclinical evaluation; preclinical study; scaffold; small molecule inhibitor; small molecule libraries; structural biology; targeted agent; targeted treatment; tumor; tumor progression

MDM2 and XIAP are important cell-survival proteins in tumor cells. MDM2 acts as an oncoprotein, promoting cancer progression mainly through inhibition of the tumor suppressor p53, while the anti-apoptotic protein XIAP plays a critical role in development of resistance to treatment via inhibition of therapy-induced apoptosis. MDM2 overexpression and upregulated XIAP have been detected in various human cancers but not in normal cells/tissues, and elevated MDM2 and XIAP expression in tumor cells is associated with disease progression and poor treatment outcomes. Our previous studies elucidated a molecular mechanism by which the MDM2 C-terminal RING domain interacts with XIAP IRES mRNA resulting in stabilization of MDM2 protein and enhanced translation of XIAP; this led to concomitantly increased expression of both MDM2 and XIAP, contributing to cancer progression and drug resistance. We have recently established a fluorescence polarization (FP) assay for use in high-throughput screening (HTS) of chemical libraries and identified a compound (MX69) that binds to the MDM2 RING domain and blocks or disrupts its interaction with XIAP IRES mRNA. Blocking this interaction results in simultaneous inhibition of both MDM2 and XIAP, leading to cancer cell apoptosis and death. The overall goal of this proposal is to develop a potential novel targeted agent based on the MX69 scaffold against tumors overexpressing MDM2. As discussed above, these tumors also typically upregulate XIAP in an MDM2-dependent manner, resulting in enhanced drug resistance. Specifically, we will perform computer-aided drug design based on the MX69 structure and iteratively optimize MDM2 binding and anticancer activity (Aim 1). We will define the molecular and biological mechanism(s) of action of novel MX69 analogs by solving the X-ray crystal structures of MDM2 in complex with diverse MX69 analogs to confirm on-target MDM2/XIAP inhibition and to evaluate any potential nonspecific effects (Aim 2). We will perform preclinical studies to assess compound stability, pharmacokinetic (PK), and pharmacodynamic (PD) properties of the best MX69 analogs; to evaluate their anticancer activity in vivo using human cancer-in-mouse models; and to determine any potential toxicity to normal cells/tissues (Aim 3). Upon completion of this project, we will have determined the feasibility of dual targeting MDM2/XIAP as a novel therapeutic mechanism, and will have developed promising small molecule inhibitors for further preclinical evaluations, not only for leukemia and neuroblastoma as studied in this proposal, but also in other cancer types.

MDM2和XIAP是肿瘤细胞中重要的细胞生存蛋白。 MDM2充当癌蛋白，促进 癌症进展主要是通过抑制肿瘤抑制p53，而抗凋亡蛋白XIAP 通过抑制治疗诱导的细胞凋亡，在对治疗的抵抗力发展中起着至关重要的作用。 MDM2 在各种人类癌症中已经检测到过表达和上调的XIAP 细胞/组织，肿瘤细胞中的MDM2和XIAP表达升高与疾病进展有关 和治疗结果不佳。 我们以前的研究阐明了MDM2 C末端环域的分子机制 与XIAP IRES mRNA相互作用，导致MDM2蛋白稳定并增强了XIAP的翻译； 这导致MDM2和XIAP的表达同时增加，导致癌症进展和 耐药性。我们最近建立了用于高通量的荧光偏振（FP）测定 化学库的筛选（HTS），并鉴定出与MDM2环域结合的化合物（MX69） 阻止或破坏其与XIAP IRES mRNA的相互作用。阻止这种相互作用导致同时 抑制MDM2和XIAP，导致癌细胞凋亡和死亡。该提议的总体目标 是基于对过表达MDM2的肿瘤的MX69支架开发潜在的新型靶向剂。 如上所述，这些肿瘤通常还以MDM2依赖性方式上调XIAP，从而导致 耐药性增强。 具体而言，我们将根据MX69结构进行计算机辅助的药物设计 优化MDM2结合和抗癌活性（AIM 1）。我们将定义分子和生物学 新型MX69类似物的作用机理，通过与MDM2的X射线晶体结构与复合物的X射线晶体结构与 多种MX69类似物以确认靶向MDM2/XIAP抑制并评估任何潜在的非特异性 效果（目标2）。我们将进行临床前研究以评估复合稳定性，药代动力学（PK）和 最佳MX69类似物的药效学（PD）特性；使用体内评估其抗癌活性 人类癌症模型；并确定对正常细胞/组织的任何潜在毒性（AIM 3）。之上 完成该项目的完成，我们将确定针对MDM2/XIAP的双重目标的可行性 治疗机制，并将开发出有希望的小分子抑制剂以进行进一步的临床前 评估，不仅是对该提案中研究的白血病和神经母细胞瘤的评估，而且对其他癌症类型进行了评估。