TY - JOUR
T1 - Hematin polymerization assay as a high-throughput screen for identification of new antimalarial pharmacophores
AU - Kurosawa, Y.
AU - Dorn, A.
AU - Kitsuji-Shirane, M.
AU - Shimada, H.
AU - Satoh, T.
AU - Matile, H.
AU - Hofheinz, W.
AU - Masciadri, R.
AU - Kansy, M.
AU - Ridley, R. G.
PY - 2000
Y1 - 2000
N2 - Hematin polymerization is a parasite-specific process that enables the detoxification of heine following its release in the lysosomal digestive vacuole during hemoglobin degradation, and represents both an essential and a unique pharmacological drug target. We have developed a high-throughput in vitro microassay of hematin polymerization based on the detection of 14C-labeled hematin incorporated into polymeric hemozoin (malaria pigment). The assay uses 96-well filtration microplates and requires 12 h and a Wallac 1450 MicroBeta liquid scintillation counter. The robustness of the assay allowed the rapid screening and evaluation of more than 100,000 compounds. Random screening was complemented by the development of a pharmacophore hypothesis using the 'Catalyst' program and a large amount of data available on the inhibitory activity of a large library of 4-aminoquinolines. Using these methods, we identified 'hit' compounds belonging to several chemical structural classes that had potential antimalarial activity. Follow-up evaluation of the antimalarial activity of these compounds in culture and in the Plasmodium berghei murine model further identified compounds with actual antimalarial activity. Of particular interest was a triarylcarbinol (Ro 06-9075) and a related benzophenone (Ro 22-8014) that showed oral activity in the murine model. These compounds are chemically accessible and could form the basis of a new antimalarial medicinal chemistry program.
AB - Hematin polymerization is a parasite-specific process that enables the detoxification of heine following its release in the lysosomal digestive vacuole during hemoglobin degradation, and represents both an essential and a unique pharmacological drug target. We have developed a high-throughput in vitro microassay of hematin polymerization based on the detection of 14C-labeled hematin incorporated into polymeric hemozoin (malaria pigment). The assay uses 96-well filtration microplates and requires 12 h and a Wallac 1450 MicroBeta liquid scintillation counter. The robustness of the assay allowed the rapid screening and evaluation of more than 100,000 compounds. Random screening was complemented by the development of a pharmacophore hypothesis using the 'Catalyst' program and a large amount of data available on the inhibitory activity of a large library of 4-aminoquinolines. Using these methods, we identified 'hit' compounds belonging to several chemical structural classes that had potential antimalarial activity. Follow-up evaluation of the antimalarial activity of these compounds in culture and in the Plasmodium berghei murine model further identified compounds with actual antimalarial activity. Of particular interest was a triarylcarbinol (Ro 06-9075) and a related benzophenone (Ro 22-8014) that showed oral activity in the murine model. These compounds are chemically accessible and could form the basis of a new antimalarial medicinal chemistry program.
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U2 - 10.1128/AAC.44.10.2638-2644.2000
DO - 10.1128/AAC.44.10.2638-2644.2000
M3 - Article
C2 - 10991837
AN - SCOPUS:0033806991
VL - 44
SP - 2638
EP - 2644
JO - Antimicrobial Agents and Chemotherapy
JF - Antimicrobial Agents and Chemotherapy
SN - 0066-4804
IS - 10
ER -