COUPLED ENZYME SYSTEMS TO PRODUCE TRYPTAMINE DERIVATIVES

生产色胺衍生物的偶联酶系统

• 批准号: 6210394
• 负责人: JAMES L KILGORE
• 金额: $ 10万
• 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: 2000-07-01 至 2000-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6210394
• 关键词: Escherichia coli; aromatic L aminoacid decarboxylase; chemical synthesis; enzyme activity; enzyme mechanism; enzyme substrate; immobilized enzymes; indoles; method development; thin layer chromatography; tryptamines; tryptophan synthase

Feasibility of a coupled-enzyme process to attach the aminoethyl group to the 3-positions of substituted indoles will be investigated. This process is offered as an alternative to existing synthetic methods, which are often not utilized in drug development for reasons discussed. Tryptophan synthase and aromatic amino acid decarboxylase will be obtained from overexpressing strains of E. coli and from pig kidney, respectively. Substrate requirements for each enzyme will be determined by qualitative screening using TLC, and quantitative enzyme activity measurements. Substituted tryptamine targets will include O-methylserotonin, 5-(hydroxymethyl)tryptamine, and key intermediates in the syntheses of selective melatonin ML1 and ML2 receptor ligands luzindole and 2-I-MCA-NAT. Additional screening of enzyme substrates will include a number of commercially-available substituted indoles and tryptophans, in order to test the substrate ranges of the individual and coupled enzyme systems. Conditions for tryptamine syntheses will be optimized for batch reactions and additional tests of 2-phase aqueous/organic solvent and immobilized enzyme reactor systems, which may improve productivity simplify product isolation, will be conducted. PROPOSED COMMERCIAL APPLICATIONS: The process proposed may provide faster access to key pharmaceutical intermediates. Costs for substituted tryptamine pharmaceutical intermediates could be reduced by an efficient route from substituted indoles. Similarly, short routes to neuroreceptor probes would increase the market for these compounds as research tools by reducing prices.

将研究偶联酶方法将氨乙基连接到取代吲哚的3-位的可行性。这个过程是作为现有合成方法的替代方法提供的，由于讨论的原因，现有合成方法通常不用于药物开发。色氨酸合酶和芳香族氨基酸脱羧酶将从E.大肠杆菌和猪肾组织中的蛋白质。将通过使用TLC进行定性筛选和定量酶活性测量来确定每种酶的底物要求。取代的色胺目标将包括O-甲基血清素，5-（羟甲基）色胺，和选择性褪黑激素ML 1和ML 2受体配体luzindole和2-I-MCA-NAT的合成中的关键中间体。酶底物的额外筛选将包括一些商业上可获得的取代吲哚和双氢吡喃，以测试个别和耦合酶系统的底物范围。色胺合成的条件将被优化用于分批反应，并将进行两相水/有机溶剂和固定化酶反应器系统的额外测试，这可能会提高生产率，简化产品分离。拟议的商业应用：拟议的工艺可以更快地获得关键的药物中间体。取代色胺药物中间体的成本可以通过取代吲哚的有效路线来降低。同样，神经感受器探针的短路线将通过降低价格来增加这些化合物作为研究工具的市场。