Fermentation initiated antibiotic synthesis

描述（由申请人提供）：Promiliad Biopharma Inc.与俄亥俄大学之间的第一阶段小企业技术转移研究，重点是对Promiliad Biopharma的核心技术REBACS进行清晰的原型演示。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，作为生物活性筛选程序的优越起点。我们对REBACS在生产新的大环内酯类抗生素化合物文库中的应用特别感兴趣。我们的总体目标是制备新的大环内酯类抗生素，克服细菌耐药机制。我们可以通过使用REBACS生成高度多样化的内酯成分来实现这一点。改性的大内酯已被证明是有效的。对大内酯进行修饰的主要限制是目前需要使用糖基化的发酵产物作为主要前体。如果发现一种去糖胺模拟物，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匹克霉素苷元匹克诺内酯偶联的模拟物库来发现这种去糖胺模拟物。我们的方法分为两个阶段，针对大环内酯和大环内酯的糖部分。我们设想了一种总体策略，其中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，这种结构不是针对生物活性进行优化，而是为了便于化学多样化。这种整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。第一阶段STTR项目的重点是展示我们项目的前两个步骤：制备小分子糖模拟物和将糖模拟物偶联到标准的，经过验证的大环内酯苷元（pikronolide），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.