Enzyme and Pathway Engineering for in vivo Production of Anticancer Noscapine Derivatives

体内生产抗癌那斯卡品衍生物的酶和途径工程

• 批准号: 9327700
• 负责人: James Thomas Payne
• 金额: $ 5.67万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9327700
• 关键词: Active Sites; Adverse effects; Alanine; Alkaloids; Anabolism; Antitussive Agents; Benzylisoquinolines; Biological; Chemicals; Clinical; Docking; Engineering; Enzymes; Escherichia coli; Fermentation; Homology Modeling; In Vitro; Investigation; Label; Malignant Neoplasms; Mentors; Mutagenesis; Nature; Noscapine; Pathway interactions; Production; Property; Protein Engineering; Recording of previous events; Research Personnel; Route; Saccharomyces cerevisiae; Testing; Training; Tyrosine; Work; Yeasts; analog; anticancer activity; combinatorial; design; drug development; enzyme biosynthesis; enzyme pathway; feeding; improved; in vivo; novel; novel therapeutics; skills; synthetic biology; tyrosine analog

PROJECT SUMMARY    Current cancer chemotherapeutics typically cause significant negative side effects;; novel chemotherapeutics  without such side effects are therefore highly sought after. The benzylisoquinoline alkaloid noscapine, which  has a long history of safe clinical use as an antitussive, is currently being investigated for anticancer properties  with far fewer side effects than current chemotherapeutics and has been prescribed off-­label for this purpose.  Use of noscapine as a cancer chemotherapeutic is hindered by low potency, but synthetic derivatives of  noscapine (noscapinoids), especially halogenated derivatives, have demonstrated significantly improved  activities. However, only a limited set of derivatives can be readily produced synthetically, and further improved  derivatives may still exist to be explored. Recently, engineered strains of Saccharomyces cerevisiae capable of  biosynthesizing noscapine have been developed, and this biosynthetic pathway could be expanded via  additional enzymes in order to provide ready access to noscapinoids, both those currently under investigation  as well as novel, potentially improved derivatives. While halogenating enzymes (halogenases) exist in nature,  no such halogenases are known to functionalize noscapine or related compounds. In this work I will utilize  protein engineering in order to generate engineered halogenases capable of functionalizing noscapine to  directly furnish halonoscapinoids. I will also engineer enzymes in the biosynthetic pathway of noscapine to  expand their substrate scopes to allow incorporation of derivatized substrates to afford access to an additional  range of noscapinoids not available via chemical or enzymatic synthesis. I will then perform pathway  engineering to combine these engineered enzymes with the heterologous noscapine biosynthetic pathway in  order to generate high value noscapinoids for biological characterization. This will ultimately furnish  biosynthetic routes to a range of noscapinoids with potential anticancer activity and will demonstrate the utility  of combining protein engineering with synthetic biology for novel drug development via combinatorial  biosynthesis. In addition, this proposal will allow me to apply and expand the skills I developed in protein  engineering during my graduate work while receiving training from my mentor and coworkers in pathway  engineering and synthetic biology in yeast.

项目总结