Discovery and development of drug cocktails evolved by Nature

项目总结 需要发现和开发新的药物和治疗替代品来 解决迅速出现的耐药性问题。一个有吸引力的方法是最大限度地发挥 目前可通过联合疗法获得的药物。然而，有效药物的从头设计 鸡尾酒因对其作用机制的有限了解或乏味的审判而受到影响- 和-错误方法。此外，不合理和滥用药物甚至会加剧耐药性。 无效的药物鸡尾酒。因此，需要一种有效的鸡尾酒配方设计策略。 大量的观察表明，微生物已经进化出大量有效的药物。 经过数十亿年的自然选择，抗击抗药性的鸡尾酒配方。然而， 除了几个已知的例子，联合生产的天然产物及其组合潜力有 几十年来一直被忽视。这主要是因为该领域的重点是 提纯的单个化合物，而不是混合物本身。此外，预测联合 以合成天然产物的微生物基因组为基础生产天然产物。此外， 合成联合生产和协同作用的天然产物仍然很困难。米拉的这笔赠款将解决 这些挑战。第一个研究方向是确定中间体和中间体之间的协同效应 由单一生物合成途径共同生产的最终抗菌天然产物。结构上的相似性 这些化合物之间的结合有望使它们与相同的靶标结合，这可能导致 对耐药微生物的协同抑制。通过表征通过生物合成的化合物 同样的途径，有望直接提供有效的抗耐药鸡尾酒配方和 加深对综合疗法设计规则的理解。第二个研究方向 是解决在生产增效化合物过程中的挑战，通过将它们的 生物合成成两个细菌宿主，一个宿主产生中间体，另一个宿主产生 最终产品。与传统的生物合成相比，它预计有三个优点 一个单一的细菌宿主。首先，这一策略将减少每个细菌种群的代谢负担 将体验，从而提高最终产品的整体生产。第二，效价比 的协同中间体和最终产品可以通过仅操纵一种细菌进行微调 主机，在优化过程中提供模块化。第三，该系统将产生化合物 与效价比表现出最佳的协同效应，进而缓解广泛的化合物分离 和净化。总体而言，这两个提出的研究方向的协同将导致这一发现 有效的抗耐药鸡尾酒配方，为设计提供了基本的见解 联合疗法的规则，并解决这些化合物生产中的挑战。

PROJECT SUMMARY Discovery and development of new pharmaceuticals and therapeutic alternatives is required to address rapid emergence of drug resistance. An attractive approach is to maximize the potential of currently available drugs through combination therapy. However, de novo design of effective drug cocktails has suffered from the limited understanding of their mechanisms of action or the tedious trial- and-error methods. Moreover, drug resistance can even be aggravated by irrational and abusive usage of noneffective drug cocktails. Therefore, a design strategy for effective cocktail recipe is needed. Numerous observations have suggested that microbes have evolved a large collection of effective drug cocktail recipes to fight against drug resistance after billions of years of natural selection. However, besides a few known examples, co-produced natural products and their combinatorial potential have been overlooked for decades. This is primarily due to the focus of the field on the biological activity of the purified individual compounds, rather than the mixture itself. In addition, it is challenging to predict co- produced natural products based on the genomes of the microbes that synthesize them. Moreover, the synthesis of co-produced and synergistic natural products remains difficult. This MIRA grant will address these challenges. The first research direction is to identify the synergies between intermediates and the final antimicrobial natural products co-produced by single biosynthetic pathways. The structural similarity between these compounds is expected to allow their binding towards the same targets, which can lead to synergistic inhibition of drug-resistant microbes. By characterizing compounds biosynthesized through the same pathways, it is expected to directly provide effective drug cocktail recipe against resistance and to deepen the understanding of the design rules for combination therapy. The second research direction is to address the challenge during the production of the synergistic compounds by dividing their biosynthesis into two bacterial hosts, with one host producing the intermediate, the other producing the final product. It is expected to have three advantages compared to the conventional biosynthesis using a single bacterial host. First, this strategy will reduce the metabolic burden that each bacterial population will experience, and thereby improve the overall production of the final products. Second, the titer ratio of the synergistic intermediates and final products can be fine-tuned by manipulating only one bacterial host, providing modularity during optimization process. Third, the system will produce the compounds with the titer ratio showing the optimal synergistic effect, and then alleviate extensive compound isolation and purification. Overall, the synergy of the two proposed research directions will lead to the discovery of effective drug cocktail recipes against drug resistance, provide fundamental insights for the design rules of combination therapy, and address challenges in the production of these compounds.