New GI-specific Histone Deacetylase Inhibitors

新型胃肠道特异性组蛋白脱乙酰酶抑制剂

• 批准号: 7004067
• 负责人: SCOTT Raymond RAJSKI
• 金额: $ 7.05万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-15 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7004067
• 关键词: Cruciferae; amidohydrolases; cadherins; cancer prevention; carboxylate; cell free system; cell line; chemical conjugate; chemoprevention; colorectal neoplasms; combinatorial chemistry; cysteine; drug design /synthesis /production; drug screening /evaluation; enzyme activity; enzyme inhibitors; gastrointestinal system; glycosides; high performance liquid chromatography; hydrolase; isothiocyanates; neoplasm /cancer pharmacology; transfection /expression vector; zinc

DESCRIPTION (provided by applicant): The primary objective of the proposed research is to develop new inhibitors of histone deacetylase (HDAC) with discrete selectivity for the human GI tract and proximal organs such as prostate, pancreas, liver and kidney. This goal is promoted by two key findings. First, colorectal cancer is the second leading cause of death from cancer in males and the third in females in the U.S. Secondly, it has been shown that glucoraphanin (GPN), the primary chemopreventive agent in broccoli undergoes myrosinase-dependent metabolism to afford a weak HDAC inhibitor. Evidence strongly suggests the GPN-derived HDAC inhibitor to be a cysteine conjugate similar in structure to C shown below. It is highly likely that the HDAC inhibiting metabolite is responsible for the beneficial chemopreventive effects associated not only with broccoli consumption but also the ingestion of many other vegetables that provide glucosinolates capable of GI-localized metabolism as shown below. Our goal is to generate new substances that share glucoraphanin's novel mechanism of action but are more potent HDAC inhibitors and thus more useful chemopreventive agents. We will first prepare a number of derivatives of the cysteine-conjugate derived from GPN to verify the hypothesis that the cysteine-derived carboxylate is responsible for Zn2+ coordination (and hence, enzyme inactivation) within the HDAC active site. From a library of primary amines, we will then prepare ITC analogs (like B shown below) and cysteine-1 inked conjugates (of type C). ITC and cysteine conjugates for each member of the library will be tested for HDAC inhibition in cell free assays (Fluor de Lys) and human colon cancer cells (HCT116). Agents that display anti-HDAC activity will be candidates for further synthetic and biochemical study. Specifically, glucosinolate precursors (of form A shown below) to these active agents will be constructed. The ability of resulting glucosinolates to undergo myrosinase-triggered conversion to the corresponding ITC will be assessed by automated reverse phase HPLC and, if applicable, we will examine the HDAC inhibition profile of said myrosinase-activated agents in both cell free assays and in HCT 116 cells.

描述（由申请人提供）：拟议研究的主要目标是开发对人类胃肠道和近端器官（如前列腺、胰腺、肝脏和肾脏）具有离散选择性的新型组蛋白脱乙酰酶（HDAC）抑制剂。两项重要发现推动了这一目标的实现。首先，结直肠癌是美国男性癌症死亡的第二大原因，女性癌症死亡的第三大原因。其次，研究表明，西兰花中的主要化学预防剂萝卜硫苷 (GPN) 会经历黑芥子酶依赖性代谢，从而提供较弱的抗癌作用。 HDAC 抑制剂。证据强烈表明 GPN 衍生的 HDAC 抑制剂是一种半胱氨酸缀合物，其结构与如下所示的 C 类似。 HDAC 抑制代谢物很可能产生有益的化学预防作用，不仅与西兰花的食用有关，而且与摄入许多其他蔬菜有关，这些蔬菜提供能够进行胃肠道局部代谢的芥子油苷，如下所示。我们的目标是产生具有萝卜硫苷新作用机制的新物质，但它们是更有效的 HDAC 抑制剂，因此是更有用的化学预防剂。我们将首先制备许多源自 GPN 的半胱氨酸缀合物衍生物，以验证半胱氨酸衍生的羧酸盐负责 HDAC 活性位点内 Zn2+ 配位（从而导致酶失活）的假设。然后，我们将从伯胺库中制备 ITC 类似物（如下所示的 B）和半胱氨酸-1 墨迹缀合物（C 型）。将在无细胞测定 (Fluor de Lys) 和人结肠癌细胞 (HCT116) 中测试文库每个成员的 ITC 和半胱氨酸缀合物的 HDAC 抑制作用。显示抗 HDAC 活性的药物将成为进一步合成和生化研究的候选药物。具体而言，将构建这些活性剂的芥子油苷前体（如下所示的形式A）。所得芥子油苷经历黑芥子酶触发转化为相应 ITC 的能力将通过自动反相 HPLC 进行评估，如果适用，我们将在无细胞测定和 HCT 116 中检查所述黑芥子酶激活剂的 HDAC 抑制谱细胞。