Androgen Receptor-JunD Complex Inhibitors to Prevent Prostate Cancer Progression

雄激素受体-JunD 复合物抑制剂可预防前列腺癌进展

• 批准号: 8315084
• 负责人: Hirak S. Basu
• 金额: $ 30万
• 依托单位: COLBY PHARMACEUTICAL COMPANY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-24 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8315084
• 关键词: Androgen Receptor; Androgens; Animal Model; Animals; Antioxidants; Apoptosis; Basic Science; Binding Sites; Bioavailable; Biological Assay; Cancer Cell Growth; Cancer Etiology; Cancer Patient; Castration; Cell Proliferation; Cessation of life; Chemicals; Clinical; Complex; Computer Assisted; Computer Simulation; Data; Development; Diet; Docking; Dose; Early treatment; Enzymes; Gene Expression; Generations; Growth; Hormones; Human; Hydrogen Peroxide; Hydroxyl Radical; In Situ; Investigation; Ionizing radiation; Lead; Legal patent; Libraries; Luciferases; Malignant Neoplasms; Malignant neoplasm of prostate; Metabolic; Metastatic Prostate Cancer; Modeling; Molecular; NF-kappa B; Nature; Nuclear; Nude Mice; Oxidative Stress; Pathway interactions; Pharmaceutical Preparations; Phase; Preclinical Testing; Process; Production; Prostate; Prostatic; Proteins; Publications; Publishing; Radiation therapy; Radical Prostatectomy; Reactive Oxygen Species; Recurrence; Refractory; Relative (related person); Reporting; Resected; Resistance; Source; Spermidine; Spermine; Staging; Superoxides; Testing; Testosterone; Tissues; Transferase; Transferase Gene; Transgenic Mice; Up-Regulation; Vitamin E; Xenograft procedure; activating transcription factor; analog; autocrine; base; cancer cell; cancer recurrence; cancer therapy; cell growth; deprivation; design; drug candidate; effective therapy; enzyme reconstitution; feeding; follow-up; high throughput screening; inhibitor/antagonist; male; member; men; mouse model; novel; overexpression; oxidation; pre-clinical; prevent; protein expression; protein protein interaction; receptor binding; research clinical testing; small molecule; therapy development; transcription factor; tumor progression

DESCRIPTION (provided by applicant): Most conventional cancer therapies are only modestly effective against CRPCa. Therefore, the development of new and effective therapies to treat early-stage prostate cancer (PCa) to prevent progression of PCa to CRPCa is warranted. Our collaborator Dr. Wilding and his coworkers' basic science investigation of probable mechanism(s) of PCa progression led to a class of natural cancer causative agents, cellular reactive oxygen species (ROS) such as superoxide, hydroxyl radical, hydrogen peroxide, etc. Their published and strong preliminary data show that in human PCa cells, excess cellular ROS activate the transcription factor NF-kappa B that prevents apoptosis and drives PCa cell proliferation in the absence of androgen. It is proposed that targeted inhibitors o PCa specific ROS generation pathway(s) will be more effective than are chemical and/or dietary anti-oxidants in preventing PCa recurrence and progression to CRPCa. Our published data show androgen induces spermidine/spermine acetyl transferase (SSAT) that initiates a spermidine and spermine oxidation pathway to generate copious amounts of ROS specifically in PCa cells that are naturally rich in spermine and spermidine. As JunD protein expression is induced by androgen specifically in PCa cells, it is hypothesized that JunD complexes with the activated androgen receptor (AR) to induce SSAT gene expression and ROS production in PCa cells. We and others have also shown that NF-kappa B can induce SSAT expression, thus setting up a feed-forward loop for SSAT activation, ROS production and PCa progression. Therefore, it is proposed that specific inhibitor(s) of JunD-AR complex should block SSAT induction, ROS production, NF-kappa B activation and stop the autocrine feed-forward loop that helps CRPCa progression. A high-throughput screen using Gaussia luciferase enzyme reconstitution assay for in situ protein-protein interaction, thus far, identified seven inhibitorsthat can block JunD-AR interaction. Two of these compounds blocked androgen-induced ROS generation in androgen-dependent PCa cells and showed growth inhibitory effects against both androgen- dependent and androgen-independent human PCa cells at sub-micromolar to low micromolar levels. Using computer-aided molecular docking to determine the binding site(s) of the inhibitor, we propose to design analogs of the two lead compounds that should more efficiently block androgen-induced ROS generation and cell growth. Our Specific Aims are: 1) To develop models for small molecule interaction with JunD- AR complex using in silico docking and design and synthesize analogs of small molecule inhibitors of JunD- AR interaction; 2) To identify promising agents from their relative abilities to disrupt JunD-AR interaction and their efficacies in reducing intracellular ROS and inhibit cultured human PCa cell growth.; 3) To identify the lead drug candidate from the studies in Aim 2 and test its ability to inhibit human PCa growth in nude mouse xenografts and the same in a transgenic mouse model developing spontaneous PCa. PUBLIC HEALTH RELEVANCE: Advanced hormone refractory metastatic prostate cancer (CRPCa) is the second leading cause of cancer deaths among US men. Most conventional cancer therapies are only modestly effective against CRPCa. Therefore, the development of new and effective therapies to prevent prostate cancer (PCa) recurrence and/or progression to CRPCa is warranted. Our collaborator Dr. Wilding and his coworkers' investigation of probable mechanism(s) of PCa recurrence and progression led to a class of natural cancer causative agents, cellular reactive oxygen species (ROS). ROS levels are generally high in PCa cells as compared to their normal counterpart. Male hormone testosterone induces copious amounts of ROS generation specifically in PCa cells. Recent discoveries show that the main source of the prostatic ROS is oxidation of spermine and spermidine that is found in very high level in the prostate. It has also been demonstrated that activated androgen receptor binds with a protein JunD to induce the spermine/spermidine oxidation. Therefore, it is proposed that specific inhibitor(s) of JunD-AR complex should inhibit spermine/spermidine oxidation and should thus block ROS production in PCa cells and prevent CRPCa proliferation. We used a high throughput screen to identify inhibitors of JunD-AR interaction and identified several compounds that can inhibit JunD-AR interaction. One of these compounds blocked androgen-induced ROS generation in androgen-dependent PCa cells and showed growth inhibitory effects against both androgen-dependent and androgen-independent human PCa cells. Colby has patented these compounds for preclinical and clinical development for treatment of early-stage progressing PCa patients. Using computer-aided molecular design to determine the binding site(s) of the inhibitor, here, we propose to develop new analogs of the lead compound that should more efficiently block androgen-induced ROS generation and cell growth. The most active agent, thus identified, can be further developed for preclinical and clinical testing to prevent progression of early-stage androgen-dependent PCa to CRPCa.

描述（由申请人提供）：大多数常规癌症疗法仅适度针对CRPCA。因此，有必要开发用于治疗早期前列腺癌（PCA）以防止PCA向CRPCA发展的新疗法。我们的合作者Wilding博士及其同事对PCA进展可能机制的基础科学研究导致一系列天然癌症致癌剂，细胞活性氧（ROS），例如超氧化物，羟基自由基，过氧化氢，其已发表和强有力的pca thate the to the to the trapection pca ros a the to the to the toca beft the toca the toca beft to the apcapa rapa beft。在没有雄激素的情况下驱动PCA细胞增殖。有人提出，靶向抑制剂O PCA特异性ROS产生途径（S）将比化学和/或饮食抗氧化剂更有效，以防止PCA复发和发展为CRPCA。我们发表的数据表明，雄激素诱导精子/精子乙酰基转移酶（SSAT），该酶启动了精子定和精子氧化途径，以在PCA细胞中特别富含精子和精子素的PCA细胞中产生大量的ROS。由于在PCA细胞中特异性雄激素诱导了JUND蛋白的表达，因此假设JUND与活化的雄激素受体（AR）诱导，以诱导PCA细胞中的SSAT基因表达和ROS产生。我们和其他人还表明，NF-kappa b可以诱导SSAT表达，从而为SSAT激活，ROS产生和PCA进展设置一个馈送回路。因此，提议Jund-AR复合物的特异性抑制剂应阻止SSAT诱导，ROS产生，NF-KAPPA B激活，并阻止自分泌馈送前回路，从而有助于CRPCA进展。使用高斯荧光素酶重构测定的高通量筛选，用于原位蛋白 - 蛋白质相互作用，到目前为止，鉴定出七个抑制剂可以阻止Jund-AR相互作用。这些化合物中的两种阻止了雄激素诱导的雄激素依赖性PCA细胞的ROS产生，并在亚微摩尔至低微摩尔水平下对雄激素依赖性和雄激素独立的人PCA细胞均显示出生长抑制作用。使用计算机辅助分子对接来确定抑制剂的结合位点，我们建议设计两种铅化合物的类似物，这些化合物应更有效地阻止雄激素诱导的ROS的产生和细胞生长。我们的具体目的是：1）开发用于与JundAR复合物相互作用的模型，并在硅离子对接和设计中使用并合成了JundAR相互作用的小分子抑制剂的类似物； 2）从其相对能力中确定有希望的药物，以破坏Jund-AR相互作用及其在减少细胞内ROS和抑制培养的人PCA细胞生长方面的效率。 3）从AIM 2中的研究中鉴定铅药物的候选者，并测试其抑制裸小鼠异种移植物中人类PCA生长的能力，而在开发自发PCA的转基因小鼠模型中相同。 公共卫生相关性：晚期激素难治性转移性前列腺癌（CRPCA）是美国男性癌症死亡的第二大原因。大多数常规的癌症疗法仅适度针对CRPCA。因此，有必要开发用于预防前列腺癌（PCA）复发和/或向CRPCA的新疗法的发展。我们的合作者Wilding博士及其同事对PCA复发和进展的可能机制的研究导致一类天然癌症剂，细胞活性氧（ROS）。与正常的对应物相比，PCA细胞中的ROS水平通常很高。雄性激素睾丸激素在PCA细胞中诱导大量的ROS产生。最近的发现表明，前列腺ROS的主要来源是在前列腺中非常高的精子和精子的氧化。还已经证明，活化的雄激素受体与蛋白质JUND结合以诱导精子/精子氧化。因此，提议Jund-AR复合物的特异性抑制剂应抑制精子/精子氧化，因此应阻止PCA细胞中的ROS产生，并防止CRPCA增殖。我们使用高吞吐量屏幕来识别Jund-AR相互作用的抑制剂，并鉴定出几种可以抑制Jund-AR相互作用的化合物。这些化合物之一阻断了雄激素诱导的雄激素依赖性PCA细胞的ROS产生，并显示出对雄激素依赖性和雄激素独立的人PCA细胞的生长抑制作用。 Colby已为临床前和临床开发申请了这些化合物，用于治疗早期进展的PCA患者。使用计算机辅助分子设计来确定抑制剂的结合位点，我们建议开发铅化合物的新类似物，这些类似物应更有效地阻止雄激素诱导的ROS的产生和细胞生长。因此，最活跃的剂可以进一步开发用于临床前和临床测试，以防止 早期雄激素依赖性PCA与CRPCA。