Chemical Approaches for the Discovery of New Cancer Therapeutic Targets

描述(申请人提供)：背景：FR901464是一种新的、有效的抗肿瘤抗生素，对小鼠和人类实体瘤都有显著的作用。FR901464在低纳摩尔浓度下强烈激活SV40驱动的报告基因，并调节人类细胞中几个癌症相关基因的表达。尽管FR901464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：深入了解FR901464的作用机制将为开发新的癌症治疗方法铺平道路。为了了解FR901464的作用机制，需要分离FR901464的靶生物分子。在这个方案中，我们的目标是化学合成FR901464及其类似物用于生物学研究，并使用放射性标记的FR901464类似物来分离目标生物分子。 具体目的：(1)利用简单的模型系统研究FR901464的化学反应活性；(2)设计和合成更稳定的FR901464类似物，将从以下两个目标进行研究；(3)确定FR901464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细胞靶点的影响；以及(4)分离并鉴定FR901464的细胞内靶点。 研究设计：首先，我们将研究FR901464在生物相关条件下的化学反应活性。在这项研究中，FR901464的每个反应部分将被用来明确地分析结果。基于从这项研究中获得的化学见解，我们随后将合成努力集中在开发稳定和有效的FR901464类似物上。这些类似物随后将在不同的癌细胞系中进行测试，以阐明FR901464的结构-活性关系(SAR)。我们还将进行高含量分析，以监测各种参数(转录活性、DNA损伤、细胞凋亡等)。这一分析将为了解FR901464及其类似物的特异性提供更深入的见解。最后，我们将制备放射性标记的FR901464类似物，在活细胞中分离FR901464的靶标。意义：FR901464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其他与这些治疗靶点结合的药物用于癌症治疗。

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.