Organosulfates and Polyphosphates: Their Synthesis and the Enzymes that Process Them.

我的研究计划的主要重点是开发新的方法来合成可用于构建具有生物重要性的化合物的分子，并评估选定的化合物作为先导结构，用于设计治疗剂和阐明具有医学意义的酶的机制。该提案的第一个主题涉及硫酸盐碳水化合物的合成。这是一类极其重要的生物分子，因为它们在许多重要的生物过程中发挥着关键作用，如血液凝结和免疫反应。我们开发了一种新的方法来制备这些化合物，它使用含有受保护的磺基双酯的单糖作为构建块。为了确定这一方法的范围，将进行区域选择性地将受保护的磺双酯加入到单糖构建块中的研究，以及受保护的磺双酯对糖基化反应的影响的研究。还将实现两种复杂的多硫酸盐低聚糖的全合成。该提案的第二部分涉及类固醇硫酸酯酶(STS)抑制剂的设计、合成和评价，STS是一种与雌激素储存有关的酶。STS的抑制剂正在作为治疗乳腺癌的治疗药物进行研究。我们已经制备了非常有效的STS可逆和不可逆抑制剂。我们将使用光亲和标记、建模、核磁共振和X射线结晶学分析来确定这些抑制剂如何与STS结合。还将对STS中基质进入和产物释放的机理进行研究。在这项建议的第三部分，将使用我们团队最近开发的一种新的、强大的磷酸化过程来构建由胞苷三磷酸合成酶(CTPS)催化的反应的中间和过渡态类似物。CTPS是抗癌和抗病毒的靶标。这些化合物将用于确定CTPS反应中哪些残基对配体结合以及中间态和过渡态的稳定是重要的。这些研究的结果将为开发针对CTPS的药物提供至关重要的信息。

The primary focus of my research program is to develop new methods for synthesizing molecules that can be used in the construction of biologically important compounds and evaluate selected compounds as lead structures for the design of therapeutic agents and for elucidating mechanisms of medicinally significant enzymes. The first subject of this proposal concerns the synthesis of sulfated carbohydrates. This is an extremely important class of biomolecules as they play key roles in numerous important biological processes such as blood clotting and the immune response. We have developed a new approach to preparing these compounds which employs monosaccharides bearing protected sulfodiesters as building blocks. To determine the scope of this methodology, studies on the regioselective incorporation of protected sulfodiesters into monosaccharide building blocks as well as the effect of protected sulfodiesters on glycosylation reactions will be performed. The total synthesis of two complex multisulfated oligosaccharides will also be achieved. The second part of this proposal deals with the design, synthesis and evaluation of inhibitors of steroid sulfatase (STS), an enzyme involved in the storage of estrogens. Inhibitors of STS are being examined as therapeutics for treating of breast cancer. We have prepared very potent reversible and irreversible inhibitors of STS. We will determine how these inhibitors bind to STS using photoaffinity labeling, modeling, NMR and x-ray crystallographic analyses. The mechanism of substrate entry and product release in STS will also be studied. In the third part of this proposal intermediate and transition state analogs of the reaction catalyzed by cytidine triphosphate synthase (CTPS), an anticancer and antiviral target, will be constructed using a novel and powerful phosphorylation procedure recently developed in our group. These compounds will be used to determine which residues are important for ligand binding and intermediate and transition state stabilization in the CTPS reaction. These results of these studies will provide information that will be crucial for developing drugs that target CTPS.