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分子，可用于构建生物学上重要的化合物，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医学上重要的酶的机制。本提案的第一个主题涉及硫酸化碳水化合物的合成。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起着关键作用，如血液凝固和免疫反应。我们已经开发了一种新的方法来制备这些化合物，它采用单糖携带受保护的硫二酯作为构建块。为了确定该方法的范围，将对受保护的硫代二酯在单糖构建块中的区域选择性结合以及受保护的硫代二酯对糖基化反应的影响进行研究。还将实现两种复合多磺酸低聚糖的全合成。本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是一种参与雌激素储存的酶。STS抑制剂正在被研究作为治疗乳腺癌的药物。我们已经制备了非常有效的可逆和不可逆STS抑制剂。我们将利用光亲和标记、建模、核磁共振和x射线晶体学分析来确定这些抑制剂如何与STS结合。本文还将研究STS中底物进入和产物释放的机理。在本论文的第三部分，我们将利用本小组最近开发的一种新的、强大的磷酸化程序，构建由抗癌和抗病毒靶点胞苷三磷酸合成酶（CTPS）催化的反应的中间态和过渡态类似物。这些化合物将用于确定在CTPS反应中哪些残基对配体结合和中间和过渡态稳定是重要的。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.