SBIR Phase I: Highly Active and Selective Hydroxylation Catalysts for Combinatorial Biocatalysis

这一小型企业创新研究(SBIR)第一阶段项目将展示使用“定向进化”创建高活性、选择性羟化酶用于潜在药物的组合生物催化的可行性。这样的蛋白质催化剂将能够直接羟化非活性碳中心，而不会氧化其他敏感的官能团；这一转化几乎不可能使用传统的化学方法。以高活性细菌酶为起点，利用高通量筛选方法，产生随机突变体，筛选出具有靶向羟基化能力的克隆。现有的细菌P450酶不具有所需的选择性。哺乳动物的P450系统在体内有效工作，很难表达，催化速度很慢。这项技术用于组合文库的潜力将通过药物支架来展示。拟议技术的商业应用最初将为开发新的候选药物提供研究氧化催化剂。将准备新的有氧考生进行测试。随后，该催化剂可用于新药的实际合成。

This Small Business Innovation Research (SBIR) Phase I project will demonstrate the feasibility of using "directed evolution" to create highly active, selective hydroxylation enzymes for use in combinatorial biocatalysis of potential drugs. Such protein catalysts will be capable of direct hydroxylation of non-activated carbon centers without oxidation of other sensitive functionality; a transformation that is virtually impossible using traditional chemistry. Using highly active bacterial enzymes as a starting point, random mutants will be generated and clones capable of targeted hydroxylations will be selected by using by using high-through-put screening methods. Existing bacterial P450 enzymes do not have the required selectivity. Mammalian P450 systems that work effectively in-vivo, are difficult to express and are slow in catalysis. The potential of this technology for combinatorial library creation will be demonstrated using drug scaffolds.The commercial application of the proposed technology will initially be to provide research oxidation catalysts for the development of new drug candidates. New classes of oxygenated candidates for testing will be prepared. Subsequently, the catalysts may be used in the actual synthesis of the new drugs.