The devastating impact of malaria includes significant mortality and illness worldwide. Increasing resistance of the causative parasite, Plasmodium, to existing antimalarial drugs underscores a need for additional compounds with distinct modes of action in the therapeutic development pipeline. Here we showcase peptide-drug conjugates (PDCs) as an attractive compound class, in which therapeutic or lead antimalarials are chemically conjugated to cell-penetrating peptides. This approach aims to enhance selective uptake into Plasmodium-infected red blood cells and impart additional cytotoxic actions on the intraerythrocytic parasite, thereby enabling targeted drug delivery and dual modes of action. We describe the development of PDCs featuring four compounds with antimalarial activity-primaquine, artesunate, tafenoquine and methotrexate-conjugated to three cell-penetrating peptide scaffolds with varied antiplasmodial activity, including active and inactive analogues of platelet factor 4 derived internalization peptide (PDIP), and a cyclic polyarginine peptide. Development of this diverse set of PDCs featured distinct and adaptable conjugation strategies, to produce conjugates with in vitro antiplasmodial activities ranging from low nanomolar to low micromolar potencies according to the drug cargo and bioactivity of the partner peptide. Overall, this study establishes a strategic and methodological framework for the further development of dual mode of action peptide-drug antimalarial therapeutics.

Keywords: Bioconjugation; Cell-penetrating peptide; Click chemistry; Malaria; Peptide-drug conjugate.

© 2024 Wiley-VCH GmbH.

疟疾的严重影响包括全球范围内显著的死亡和疾病。导致疟疾的寄生虫恶性疟原虫（Plasmodium）对现有抗疟药物的耐药性增加，强调了需要在治疗开发管道中添加具有不同作用机制的新化合物。在这里，我们展示了肽-药物偶联物（PDCs）作为一类有吸引力的化合物，在这类化合物中，治疗或潜在的抗疟疾药物通过化学方法与细胞穿透肽连接在一起。这一方法旨在增强其选择性进入受感染的红细胞，并赋予对胞内寄生虫的额外细胞毒性作用，从而实现靶向药物递送和双重作用机制。我们描述了开发了一系列PDCs，其中包含四种具有抗疟活性的化合物——伯氨喹、青蒿素、塔法诺喹和甲氨蝶呤，这些化合物与三种不同抗疟活性的细胞穿透肽支架连接在一起，包括血小板因子4衍生内化肽（PDIP）及其活性和非活性类似物，以及环状多精氨酸肽。这一系列PDCs的不同且可调适的偶联策略产生了具有体内抗疟活性、根据药物载荷和伙伴肽生物活性从低纳摩尔到低微摩尔效力的一系列共轭体。总体而言，这项研究为双重作用机制的肽-药物抗疟疾治疗进一步开发建立了一个战略和方法框架。

关键词：生物连接；细胞穿透肽；点击化学；疟疾；肽-药物偶联物。