Production of Chiral Aminoalcohols

描述(申请人提供)：在第一阶段，酶还原步骤 建立了手性邻氨基醇的立体选择性合成方法。 评估了8种不同的酮还原酶，其中包括5种酮还原酶 在这个项目中被克隆和表达，这些酶被 以底物和立体化学偏好为特征。减少 2-取代-β-酮酯，手性氨基醇的关键前体， 给出了高对映选择性和非对映选择性的产物。再循环 烟酰胺辅因子的数量超过了5000，导致 有利的经济预测。在第2阶段中，我们将创建 通过诱变克隆的基因来立体选择性的酮还原酶，使 广泛结构的β-酮酯的立体选择性还原 射程。一种克隆宿主将被开发出来，它不需要抗生素 对质粒保持和产生酮还原酶的抗性 将在实验室规模的发酵罐中进行演示。反应条件将 使用双水相有机体系进行优化，以最大限度地提高体积 在水中的溶解度有限的酮的生产率。化学物质 将演示生产氨基醇产品的重排，以及 基于使用金属亲和树脂的程序将被开发用于 从反应环境中回收氨基醇产品。三 具有重要商业价值的手性邻氨基醇将在 多克等量展示了整体技术。 建议的商业应用： 生产现有药物和新药的关键中间体的生产 产品。

DESCRIPTION (provided by applicant): In Phase 1, the enzymatic reduction step for stereoselective synthesis of chiral vicinal aminoalcohols was established. Eight different ketoreductases were evaluated, including 5 ketoreductases that were cloned and expressed during this project, and the enzymes were characterized for substrate and stereochemical preferences. Reduction of 2-substituted-beta-ketoesters, the key precursors for chiral aminoalcohols, gave products of high enantioselectivity and diastereoselectivity. Recycle numbers for nicotinamide cofactors of more than 5000 were achieved, leading to favorable economic projections. In Phase 2 we will create a library of stereoselective ketoreductase enzymes by mutagenizing cloned genes, enabling the stereoselective reduction of beta-ketoesters spanning a broad structural range. A cloning host will be developed that eliminates the need for antibiotic resistance for plasmid maintenance, and production of ketoreductase enzymes will be demonstrated in laboratory-scale fermentors. Reaction conditions will be optimized using two-phase aqueous-organic systems to maximize volumetric productivity for ketones that have limited solubility in water. The chemical rearrangement to produce the aminoalcohol products will be demonstrated, and a procedure based on the use of metal-affinity resins will be developed for recovering the aminoalcohol products from the reaction milieu. Three commercially important chiral vicinal aminoalcohols will be produced in multi-gram amounts to demonstrate the overall technology. PROPOSED COMMERCIAL APPLICATION: Production of key intermeidates for the production of existing and new pharmaceutical products.