Fermentation initiated antibiotic synthesis

描述(由申请人提供)：Promiliad Biophma Inc.和俄亥俄大学之间的这项第一阶段小型企业技术转移研究重点是Promiliad Biophma的核心技术REBACS的清晰原型演示。重组生物技术辅助组合合成(REBACS)是利用合成和/或遗传方法构建高复杂性、高多样性的化合物文库，作为生物活性筛选计划的良好起点。我们对REBAC在生产新的大环内酯类抗生素化合物文库方面的应用特别感兴趣。我们的总体目标是制备新的大环内酯类抗生素，以克服细菌耐药机制。我们可以通过使用REBAC来产生高度多样化的大内酯成分来实现这一点。大内酯的改性已被证明是有效的。对大内酯进行修饰的主要限制是目前需要将糖基化发酵产物作为主要前体。如果发现地索胺模拟物，可以设想对大内酯进行更广泛的修饰。因此，我们的首要任务是通过分析与已证实的吡克罗霉素苷元吡克罗内酯偶联的模拟文库来发现这样的地索胺模拟物。我们的方法被分成两个阶段，分别针对大内酯和大环内酯的糖部分。我们设想了一个总体战略，其中对抗生素生物合成基因的遗传操作将首先产生一种新的大环内酯结构，该结构不是为了生物活性而是为了便于化学多样化而优化的。这一总体方法的基础是假设我们将使用的糖模拟物将提供具有生物活性的新大环内酯类化合物。这个第一阶段的STTR项目集中展示了我们计划的前两步：制备小分子糖模拟物和将糖模拟物连接到标准的、经过验证的大环内酯苷元(匹克罗内酯)，并获得具有抗菌活性的化合物。

DESCRIPTION (provided by applicant): This Phase I Small Business Technology Transfer study between Promiliad Biopharma Inc. and Ohio University is focused on a clear prototype demonstration of REBACS, the core technology of Promiliad Biopharma. REcombinant Biotechnology Assisted Combinatorial Synthesis (REBACS) is the use of synthetic and/or genetic approaches to the construction of high complexity, high diversity compound libraries that serve as a superior starting point for biological activity screening programs. We have been particularly interested in the application of REBACS for the production of libraries of new macrolide antibiotic compounds. Our overall objective is to prepare new macrolide antibiotics that overcome bacterial resistance mechanisms. We can do this by using REBACS to generate highly diverse macrolactone components. Modifications of the macrolactone have been demonstrated to be effective. The major limitation in modifications to the macrolactone is the current need to work with a glycosylated fermentation product as the primary precursor. Much more extensive modifications to the macrolactone could be envisaged if a desosamine mimetic was found. Our first priority, therefore, is to discover such a desosamine mimetic by analysis of a library of mimetics coupled to the proven pikromycin aglycon, pikronolide. Our approach is divided into 2 phases targeted both at the macrolactone and the sugar portions of the macrolide. We envisage an overall strategy wherein genetic manipulation of the antibiotic biosynthesis genes will first generate a new macrolide structure that is optimized not for biological activity, but for ease of chemical diversification. The base upon which this overall approach stands is the assumption that the sugar mimetics that we will use will provide biologically active new macrolides. This Phase I STTR project is focused on demonstrating the first 2 steps of our program: Preparation of small molecule sugar mimetics and conjugation of the sugar mimetics to a standard, proven macrolide aglycon (pikronolide) and obtain compounds with antibacterial activity.