Production of Chiral Aminoalcohols

手性氨基醇的生产

• 批准号: 6401842
• 负责人: JAMES DAVID ROZZELL
• 金额: $ 50.02万
• 依托单位: BIOCATALYTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-02-01 至 2003-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6401842
• 关键词: affinity chromatography; aldehyde /ketone oxidoreductase; aminoalcohol; chemical registry /resource; chemical synthesis; cofactor; combinatorial chemistry; drug design /synthesis /production; fungal proteins; high performance liquid chromatography; immobilized enzymes; nicotinamide; plasmids; stereochemistry; stereoisomer; technology /technique development; water solubility

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.

描述（由申请方提供）：在第1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，包括5种酮还原酶， 在这个项目中克隆并表达了这些酶， 其特征在于底物和立体化学偏好。减少 2-取代-β-酮酯，手性氨基醇的关键前体， 得到了高的对映体选择性和非对映体选择性的产物。回收 烟酰胺辅因子的数量超过5000，导致 有利的经济预测。在第二阶段，我们将创建一个 立体选择性酮还原酶通过诱变克隆基因， β-酮酯的立体选择性还原跨越广泛的结构 范围一种克隆宿主将被开发出来，它不需要抗生素 对质粒维持和酮还原酶产生的抗性 将在实验室规模的发酵罐中进行演示。反应条件将 使用两相水-有机系统进行优化， 对于在水中具有有限溶解度的酮的生产率。化学 重排生产氨基醇产品将被证明，和 将开发基于使用金属亲和树脂的程序， 从反应环境中回收氨基醇产物。三 商业上重要的手性邻位氨基醇将在 几克的量来展示整个技术。 拟定商业应用： 生产用于生产现有和新药物的关键中间体 产品.