Mode of action of quinoline antimalarial drugs in red blood cells infected by Plasmodium falciparum revealed in vivo

• Published in: Proceedings of the National Academy of Sciences - PNAS 2019-11, Vol.116 (46), p.22946-22952
• Main Authors: Kapishnikov, Sergey, Staalsø, Trine, Yang, Yang, Lee, Jiwoong, Pérez-Berná, Ana J., Pereiro, Eva, Werner, Stephan, Guttmann, Peter, Leiserowitz, Leslie, Als-Nielsen, Jens
• Format: Article
• Language: English
• Subjects: Antimalarials - pharmacology; Biocompatibility; Biological Sciences; Blood; Cell culture; Chloroquine; Crystal growth; Crystal surfaces; Crystallization; Crystals; Detoxification; Drugs; Erythrocytes; Erythrocytes - chemistry; Erythrocytes - metabolism; Erythrocytes - parasitology; Heme; Heme - chemistry; Heme - metabolism; Hemeproteins - chemistry; Hemeproteins - metabolism; Hemoglobin; Hemozoin; Humans; Malaria, Falciparum - drug therapy; Malaria, Falciparum - metabolism; Malaria, Falciparum - parasitology; Mapping; Membranes; Mode of action; Parasites; Physical Sciences; Plasmodium falciparum; Plasmodium falciparum - drug effects; Plasmodium falciparum - physiology; Quinoline; Quinolines - pharmacology; Spillage; X ray microscopy
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.1910123116

The most widely used antimalarial drugs belong to the quinoline family. Their mode of action has not been characterized at the molecular level in vivo. We report the in vivo mode of action of a bromo analog of the drug chloroquine in rapidly frozen Plasmodium falciparum-infected red blood cells. The Plasmodium parasite digests hemoglobin, liberating the heme as a byproduct, toxic to the parasite. It is detoxified by crystallization into inert hemozoin within the parasitic digestive vacuole. By mapping such infected red blood cellswith nondestructive X-ray microscopy, we observe that bromoquine caps hemozoin crystals. The measured crystal surface coverage is sufficient to inhibit further hemozoin crystal growth, thereby sabotaging heme detoxification. Moreover, we find that bromoquine accumulates in the digestive vacuole, reaching submillimolar concentration, 1,000-fold more than that of the drug in the culture medium. Such a dramatic increase in bromoquine concentration enhances the drug’s efficiency in depriving heme from docking onto the hemozoin crystal surface. Based on direct observation of bromoquine distribution in the digestive vacuole and at its membrane surface, we deduce that the excess bromoquine forms a complex with the remaining heme deprived from crystallization. This complex is driven toward the digestive vacuole membrane, increasing the chances of membrane puncture and spillage of heme into the interior of the parasite.