High throughput chemoenzymatic synthesis of antimalarial compounds

项目概要 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。这些令人震惊的数字说明了这种寄生虫病的严重程度，尽管 预防方案，例如经过杀虫剂处理的蚊帐和可用的药物。因此，对抗这种疾病将 需要采取多管齐下的方法，包括疫苗等预防措施和尽量减少感染的策略 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 即将取得成功，现有小分子药物的疗效受到指数增长的威胁 恶性疟原虫（P. falciparum）的耐药性。例如，氯喹耐药性稳步上升 这种药物于 1945 年首次推出，随着其广泛分布而传播，并且已经出现了耐药性 基于青蒿素的联合疗法 (ACT) 首次于 2000 年代中期实施。根据这个轨迹，有 迫切需要开发新型小分子抗疟药物。 从历史上看，天然植物代谢物为有效的抗疟药物提供了基础。例如， 奎宁是第一种从金鸡纳树皮中分离出来的抗疟药，它提供了结构 历史上使用最广泛的小分子疟疾治疗药物氯喹的基础。同样，青蒿素是 也是一种植物天然产品，已成为治疗恶性疟疾的首选方法，并且经常被 与不同类别的抗疟药物（包括甲氟喹和阿莫地喹）一起服用。的 天然产物（如青蒿素）的复杂结构可能会使整个过程变得复杂甚至完全脱轨。 开发具有前景的抗疟活性的天然代谢物作为治疗剂。这留下了强有力的 抗疟原虫天然产物尚未得到充分开发。例如，植物天然产物，如 萘基异喹啉生物碱和芳基蒽醌，显示出对抗恶性疟原虫的效力和选择性 对于人体细胞系上的寄生虫，其效果超过氯喹，但对天然化合物的获取受到限制 和合成类似物限制了这些化合物作为药物的开发。 克服与构建复杂联芳天然产物相关的挑战，例如 通过传统合成萘基异喹啉生物碱和芳基蒽醌，我们提出了一种收敛方法 针对此类分子的化学酶方法。相关生物合成途径 萘基异醌和芳基蒽醌植物代谢物尚未被鉴定，因此，我们设想 为此目的，采用与细菌和真菌生物合成途径相关的酶。我们将聘用 高通量蛋白质工程策略，用于调整并确保获得具有合适底物的酶 范围和所需的位点选择性和天体选择性。能够快速生成有针对性的天然产品类别 及其类似物，我们将准备好询问有助于其功效和作用的结构要素 优化这些化合物作为下一代抗疟药。

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.