High throughput chemoenzymatic synthesis of antimalarial compounds

项目总结 疟疾继续对人类健康构成重大威胁，每年有超过2亿病例，近 全球有50万人死亡。这些惊人的数字说明了这种寄生虫病的严重性，尽管 预防方案，如驱虫蚊帐和可获得的药物。因此，与这种疾病作斗争将 需要多管齐下的方法，包括预防措施，如疫苗和战略，以尽量减少 传播以及有效的治疗。尽管疟疾疫苗和社会项目的最新进展 ，现有的小分子药物的疗效受到了指数级增长的威胁 恶性疟原虫耐药性的研究例如，对氯喹的抵抗力稳定 随着1945年首次引入这种药物的广泛分布而传播，已经出现了对 青蒿素为主的综合疗法于2000年中期首次应用于S。根据这一轨迹，有 迫切需要开发新型小分子抗疟疾药物。 从历史上看，天然植物代谢物为有效的抗疟疾药物提供了基础。例如, 奎宁是第一种从金鸡纳树皮中分离出来的抗疟疾药物，它提供了结构上的 氯喹是历史上使用最广泛的小分子疟疾治疗药物。同样，青蒿素也是 也是一种植物天然产品，已成为治疗恶性疟疾的首选药物，经常被 与不同类别的抗疟疾药物一起使用，包括甲氟喹和阿莫地喹。这个 天然产物的复杂结构，如青蒿素，可能会使 开发具有良好抗疟疾活性的天然代谢物作为治疗剂。这留下了强大的力量 抗疟原虫天然产物开发不足。例如，植物天然产品，如 萘基异喹啉生物碱和芳基蒽醌类化合物对恶性疟原虫具有较强的杀灭活性和选择性 对于人体细胞系上的寄生虫，超过氯喹，但限制了对天然化合物的访问 合成类似物限制了这些化合物作为药物的发展。 为了克服与构建复杂的联芳基天然产物相关的挑战，如 通过传统的合成方法，我们提出了一种收敛的 对这类分子的化学酶促方法。与之相关的生物合成途径 目前尚未鉴定出植物代谢物 为此目的调整与细菌和真菌生物合成途径相关的酶。我们将聘用 一种高通量蛋白质工程策略，以调整并确保获得具有合适底物的酶 范围和所需位置--以及可选性。能够快速生成目标天然产品类别 和它们的类比，我们将准备好询问有助于它们的有效性和 将这些化合物优化为下一代抗疟疾药物。

PROJECT SUMMARY Malaria continues to pose a significant threat to human health with over 200 million cases each year and nearly 500,000 deaths worldwide. These staggering numbers illustrate the magnitude of this parasitic disease despite preventive programs such as insecticide-treated nets and available drugs. Thus, combatting this disease will require a multiprong approach that includes preventative measures such as vaccines and strategies to minimize transmission as well as effective treatments. Although recent advances in a malaria vaccine and social programs are poised for success, the efficacy of available small molecule drugs is threatened by the exponential rise in drug-resistance in Plasmodium falciparum (P. falciparum). For example, chloroquine resistance has steadily spread with the broad distribution of this drug first introduced in 1945 and resistance has already emerged toward artemisinin-based combination therapy (ACT) first administered in the mid-2000's. Based on this trajectory, there is a dire need for the development of novel small molecule antimalarial drugs. Historically, natural plant metabolites have provided the basis for potent antimalarial drugs. For example, quinine, the first antimalarial drug which is isolated from the bark of the Cinchona tree, provided the structural basis for the most widely used small molecule malaria treatment in history, chloroquine. Similarly, artemisinin is also a plant natural product, which has become the favored treatment for falciparum malaria and is often administered along with different classes of antimalarial drugs including mefloquine and amodiaquine. The complex structures of natural products, like artemisinin, can complicate or even completely derail the development of natural metabolites with promising antimalarial activity as therapeutic agents. This has left potent antiplasmodial natural products underexplored. For example, plant natural products, such as naphthylisoquinoline alkaloids and aryl anthraquninones, display potency against P. falciparum and selectivity for the parasite over human cell lines that exceeds chloroquine, yet restricted access to the natural compounds and synthetic analogs has limited the development of these compounds as medicines. To overcome the challenges associated with constructing complex biaryl natural products such as naphthylisoquinoline alkaloids and aryl anthraquinones through traditional synthesis, we propose a convergent chemoenzymatic approach toward this class of molecules. The biosynthetic pathways related to naphthylisoquinone and aryl anthraquinone plant metabolites have not been identified, therefore, we envision adapting enzymes associated with bacterial and fungal biosynthetic pathways for this purpose. We will employ a high throughput protein engineering strategy to tune and ensure access to enzymes with suitable substrate scope and desired site- and atroposelectivity. With the ability to rapidly generate targeted natural product classes and analogs thereof, we will be poised to interrogate the structural elements that contribute to their efficacy and optimized these compounds as next-generation antimalarial drugs.