Coupled Enzyme Process for Tryptamine Synthesis

色胺合成的偶联酶工艺

• 批准号: 6442844
• 负责人: JAMES L KILGORE
• 金额: $ 43.21万
• 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 2004-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6442844
• 关键词: Escherichia coli; aromatic L aminoacid decarboxylase; chemical synthesis; directed evolution; enzyme activity; enzyme mechanism; enzyme substrate; immobilized enzymes; indoles; melatonin; method development; sumatriptan; thin layer chromatography; tryptamines; tryptophan synthase

DESCRIPTION (provided by applicant): Neuroactive tryptamine derivatives can be prepared by a new coupled-enzyme process developed in Phase 1 by combining tryptophan-synthetic and -decarboxylating enzymes. This comprises the first reported single-pot aminoethylation process. The tryptamine products can thereby be conveniently produced from readily-available indole precursors. Expression systems which provide higher enzyme yields will be utilized in order to permit scaleup to preparative reaction systems. The gene encoding Camptothectin accuminata tryptophan decarboxylase 1 (TDC 1) will be redesigned to optimize expression in E. coli. Stepwise binding of tryptophan synthase subunits and mutation of specific sites on the enzyme surface in order to improve enzyme activity following immobilization. Directed evolution will be explored in an effort to broaden the substrate ranges of two enzymes. New substrates will be tested he process will be utilized in the preparation of the migraine drugs rizatriptan, and sumatriptan, as well as melatonin and a series of melatonin receptor active compounds. PROPOSED COMMERCIAL APPLICATION: The coupled aminoethylation process will be used to make intermediates for neuropharmaceuticals, including both currently-approved and developmental neuroactive drugs.

描述（由申请人提供）：神经活性色胺衍生物可以是 通过第一阶段开发的新的偶联酶工艺制备，将 色氨酸合成酶和色氨酸脱羧酶。这包括第一个 报道了单锅氨乙基化工艺。色胺产品可以 从而可以由容易获得的吲哚前体方便地制备。 将利用提供更高酶产量的表达系统 以允许放大到制备反应系统。基因编码 喜树碱色氨酸脱羧酶 1 (TDC 1) 将被重新设计 优化大肠杆菌中的表达。色氨酸合酶的逐步结合 亚基和酶表面特定位点的突变，以便 固定化后提高酶活性。定向进化将是 旨在扩大两种酶的底物范围。新的 将测试基材，该工艺将用于制备 偏头痛药物利扎曲坦、舒马曲坦，还有褪黑素等一系列 褪黑激素受体活性化合物。 拟议的商业应用： 耦合氨乙基化工艺将用于制造中间体 神经药物，包括目前批准的和正在开发的 神经活性药物。