Chemical Approaches for the Discovery of New Cancer Therapeutic Targets

描述（由申请人提供）：背景：FR 901464是一种新型强效抗肿瘤抗生素，对鼠和人实体瘤均表现出显著作用。FR 901464在低纳摩尔浓度下强烈激活SV 40驱动的报告基因，并调节人类细胞中几种癌症相关基因的表达。尽管FR 901464具有独特的生物活性，但其生物学机制仍有待阐明。目的/假设：对FR 901464作用机制的深入了解将为开发新的癌症治疗方法铺平道路。为了了解FR 901464的作用模式，需要分离FR 901464的靶生物分子。在这个提议中，我们的目标是化学合成FR 901464及其类似物用于生物学研究，并使用放射性标记的FR 901464类似物来分离目标生物分子。 具体目标：（1）利用简单的模型系统研究FR 901464的化学反应性;（2）设计合成更稳定的FR 901464类似物，主要研究两个方面：（3）确定FR 901464类似物对人肿瘤细胞系和正常细胞的特异性，并通过高含量分析研究其对转录调控的细胞靶点的作用;和（4）分离和表征FR 901464的细胞内靶点。 研究设计：首先，我们将在生物学相关条件下研究FR 901464的化学反应性。在本研究中，FR 901464的每个反应性部分将用于明确分析结果。基于从这项研究中获得的化学见解，我们将重点放在开发稳定和有效的FR 901464类似物上。然后将这些类似物与各种癌细胞系进行测试，以阐明FR 901464的结构-活性关系（SAR）。我们还将进行高内容分析，以监测各种参数（转录活性，DNA损伤，细胞凋亡等）。该分析将为FR 901464及其类似物的特异性提供更先进的见解。最后，我们将制备放射性标记的FR 901464类似物，以分离活细胞中FR 901464的靶点。重要性：FR 901464靶点的分离将对抗癌药物的开发产生重大影响，因为我们预计几家制药公司将开始寻找结合这些治疗靶点的其他药物用于癌症治疗。

DESCRIPTION (provided by applicant): Background: FR901464 is a novel and potent antitumor antibiotic that exhibits prominent effects against both murine and human solid tumors. FR901464 strongly activates an SV40-driven reporter gene at low nanomolar concentrations and regulates the expression of several cancer-related genes in human cells. Despite the unique and promising biological activity of FR901464, its biological mechanisms remain to be elucidated. Objective/Hypothesis: A firm understanding of the mode of action of FR901464 will pave the way for the development of a new approach for cancer therapy. To understand the mode of action of FR901464, the target biomolecules of FR901464 need to be isolated. In this proposal, we aim to chemically synthesize FR901464 and its analogs for biological studies and use a radio-labeled FR901464 analog to isolate the target biomolecules. Specific Aims: (1) To study the chemical reactivity of FR901464 by using simple model systems; (2) to design and synthesize more stable FR901464 analogs, which will be studied in following two aims; (3) to determine the specificity of FR901464 analogs toward human tumor cell lines and normal cells and to investigate their effects on transcriptionally regulated cellular targets by high-content analysis; and (4) to isolate and characterize intracellular targets of FR901464. Study Design: First, we will study the chemical reactivities of FR901464 under biologically relevant conditions. In this study, each reactive moiety of FR901464 will be used to unambiguously analyze the results. Based on the chemical insight gained from this study, we then focus our synthetic efforts on developing stable and potent FR901464 analogs. These analogs will then be tested with various cancer cell lines to elucidate the structure-activity relationships (SAR) of FR901464. We will also perform high-content analysis to monitor various parameters (transcriptional activity, DNA damage, apoptosis, etc.). This analysis will provide more advanced insight into the specificity of FR901464 and its analogs. Finally, we will prepare radio-labeled FR901464 analogs to isolate the targets of FR901464 in live cells. Significance: The isolation of the targets of FR901464 will have significant impacts on anticancer drug development, as we envision several pharmaceutical companies will begin to look for other drugs that bind these therapeutic targets for cancer treatment.