Use of transaminase enzymes for the synthesis of pharmaceutical intermediates

转氨酶在药物中间体合成中的用途

• 批准号: BB/H016589/1
• 金额: $ 10.61万
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Training Grant
• 财政年份: 2010
• 资助国家: 英国
• 起止时间: 2010 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FH016589%2F1
• 关键词: Use; transaminase; enzymes; synthesis; pharmaceutical

The use of enzymes and synthetic biology strategies hold significant potential for the synthesis of pharmaceutical intermediates, and sustainable syntheses of chiral amines are highly sought after since 70% of all pharmaceutical are derivatives of chiral amines. To date chiral amines have been generated using biocatalytic strategies predominantly via the kinetic resolution of racemic mixtures with hydrolytic enzymes, where only a 50% maximum yield can be achieved. A deracemization biocatalytic strategy has also been described [1,2], however, recently interest has focused of the use of transaminases (TAm) to generate chiral amines in a genuinely asymmetric transformation [3]. This has the potential for a process to be developed using transaminases that would yield 100% of the asymmetric product, resulting in less waste and a lower cost strategy to these important synthons. TAms catalyse the transfer of an amino group from a donor such as an amino acid, to an acceptor ketone or aldehyde moiety. While the alpha-TAms have a strong preference for an alpha-keto acid as the acceptor and the preferred donor is usually one of the 20 alpha-amino acids, the omega-transaminases can transfer an amino group to an aldehyde or a ketone and do not frequently have a requirement for the alpha-keto acid moiety [4]. They also have a broader range of amino donors that they can use. In previous work by Shin and co-workers [5] an omega-TAm used for the amination of a wide range of ketones and aldehydes, including aromatic substrates, was isolated from Vibrio fluvialis and we used the protein sequence of V. fluvialis JS17 omega-TAm to screen the genome databases for related enzymes [6]. This bioinformatics approach yielded several omega-TAms including one from Chromobacterium violaceum DSM30191 that can convert of a range of ketones and aldehydes, including aliphatic and aromatic 1,3-dihydroxy ketones, resulting in very high stereoselectivities in the product ((S)-amine) [6]. In a current, 12 month EPSRC Follow-on-Fund award (EP/G005834/1), this bioinformatics strategy to identify new TAms is being pursued to find further TAms that readily convert a range of aliphatic, cyclic and aromatic ketones and aldehydes. Our aim with a BBSRC industrial CASE project is to extend the bioinformatics screen to new groups of TAms and then use several of the TAms from the EPSRC project, and new TAms (including for example sugar-specific TAms and the omega, beta and gamma-TAms) which will be cloned and characterised as part of the project, for the synthesis of chiral amines of interest to the collaborating company Chirotech Technology Limited. This will enable application of the TAm biotransformation strategy in an industrial environment and establish key advantages and problems of translating this approach at a larger scale. References: [1] Turner, N.J. Curr. Opin. Biotechnol., 2003, 14, 401. [2] Pàmies, O.; Bäckvall, J.E. Trends Biotechnol., 2004, 22, 130. [3] Koszelewski, D.; Clay, D.; Rozzell, D.; Kroutil, W. Eur. J. Org. Chem., 2009, 2289. [4] Hwang, B.Y.; Byung-Kwan, C. B. K.; Yun, H.; Kinera, K. K.; Kim, J. Mol. Catal. B: Enz., 2005, 37, 47. [5] Shin, J.S.; Kim, B.G. Biosci. Biotechnol. Biochem., 2001, 65, 1782 [6] Kaulmann, U.; Smithies, K.; Smith, M.E.B.; Hailes, H.C.; Ward, J.M.; Enzyme Microb. Technol., 2007, 41, 628.

酶和合成生物学策略的使用为药物中间体的合成提供了巨大的潜力，并且手性胺的可持续合成受到高度追捧，因为所有药物的70%是手性胺的衍生物。迄今为止，手性胺已经使用生物催化策略主要通过用水解酶动力学拆分外消旋混合物来产生，其中仅可以实现50%的最大产率。也已经描述了去外消旋化生物催化策略[1，2]，然而，最近的兴趣集中在使用转氨酶（TAm）在真正的不对称转化中产生手性胺[3]。这有可能开发一种使用转氨酶的方法，该方法将产生100%的不对称产物，从而减少浪费并降低这些重要的底物的成本。TAms催化氨基从供体如氨基酸转移到受体酮或醛部分。虽然α-TAm强烈偏好α-酮酸作为受体，并且优选的供体通常是20种α-氨基酸中的一种，但ω-转氨酶可以将氨基转移到醛或酮上，并且通常不需要α-酮酸部分[4]。它们也有更广泛的氨基供体可供使用。在Shin及其同事的先前工作中[5]，从河流弧菌中分离出一种用于胺化各种酮和醛（包括芳香底物）的omega-TAm，我们使用河流弧菌JS 17 omega-TAm的蛋白质序列筛选基因组数据库中的相关酶[6]。这种生物信息学方法产生了几种ω-TAm，包括来自紫色色杆菌DSM 30191的一种，其可以转化一系列酮和醛，包括脂肪族和芳香族1，3-二羟基酮，导致产物（（S）-胺）中非常高的立体选择性[6]。在目前为期12个月的EPSRC后续基金奖（EP/G 005834/1）中，正在寻求这种识别新TAms的生物信息学策略，以发现易于转化一系列脂肪族，环状和芳香族酮和醛的其他TAms。我们的BBSRC工业CASE项目的目标是将生物信息学筛选扩展到新的TAm组，然后使用来自EPSRC项目的几种TAm，以及将作为项目的一部分克隆和表征的新TAm（包括例如糖特异性TAm和omega，beta和gamma-TAm），用于合成合作公司Chirotech Technology Limited感兴趣的手性胺。这将使TAm生物转化策略在工业环境中的应用成为可能，并建立了在更大规模上转化这种方法的关键优势和问题。参考文献：[1] Turner，N. J. Curr. Opin.生物技术，2003，14，401。[2]Pàmies，O.; Bäckvall，J.E. Trends Biotechnol.，2004年，第22页，第130页。[3]Koszelewski，D.; Clay，D.; Rozzell，D.; Kroutil，W. EUR.有机化学杂志，2009年，2289年。[4]Hwang，B.Y.; Byung-Kwan角B。K.的; Yun，H.;基内拉，K. K.的; Kim，J. Mol.催化剂B：Enz.，2005年，37，47。[5]Shin，J.S.; Kim，B.G. Biosci. Biotechnol.生物化学，2001，65，1782 [6] Kaulmann，U.;史密斯，K.;史密斯，M.E.B.; Hailes，H.C.; Ward，J.M.;酶微生物技术人员：2007，41，628.