In vitro Plasmodium falciparum drug sensitivity assay: inhibition of parasite growth by incorporation of stomatocytogenic amphiphiles into the erythrocyte membrane.
Lupeol, which shows in vitro inhibitory activity against Plasmodium falciparum 3D7 strain with a 50% inhibitory concentration (IC50) of 27.7 +/- 0.5 microM, was shown to cause a transformation of the human erythrocyte shape toward that of stomatocytes. Good correlation between the IC50 value and the membrane curvature changes caused by lupeol was observed. Preincubation of erythrocytes with lupeol, followed by extensive washing, made the cells unsuitable for parasite growth, suggesting that the compound incorporates into erythrocyte membrane irreversibly. On the other hand, lupeol-treated parasite culture continued to grow well in untreated erythrocytes. Thus, the antiplasmodial activity of lupeol appears to be indirect, being due to stomatocytic transformation of the host cell membrane and not to toxic effects via action on a drug target within the parasite. A number of amphiphiles that cause stomatocyte formation, but not those causing echinocyte formation, were shown to inhibit growth of the parasites, apparently via a mechanism similar to that of lupeol. Since antiplasmodial agents that inhibit parasite growth through erythrocyte membrane modifications must be regarded as unsuitable as leads for development of new antimalarial drugs, care must be exercised in the interpretation of results of screening of plant extracts and natural product libraries by an in vitro Plasmodium toxicity assay.
Ziegler
Hanne L
HL
Department of Medicinal Chemistry, The Royal Danish School of Pharmacy, Copenhagen, Denmark.
Staerk
Dan
D
Christensen
Jette
J
Hviid
Lars
L
Hägerstrand
Henry
H
Jaroszewski
Jerzy W
JW
D000818:Animals; D002462:Cell Membrane / Q000187:drug effects; D004912:Erythrocytes / Q000187:drug effects* / Q000469:parasitology; D006801:Humans; D016778:Malaria, Falciparum / Q000469:parasitology; D021261:Parasitic Sensitivity Tests; D053978:Pentacyclic Triterpenes; D010963:Plasmodium falciparum / Q000187:drug effects*; D013501:Surface-Active Agents / Q000737:chemistry / Q000494:pharmacology*; D014315:Triterpenes / Q000737:chemistry / Q000494:pharmacology*; D029649:Violaceae / Q000737:chemistry; D000094042:Lupanes
D053978:Pentacyclic Triterpenes; D013501:Surface-Active Agents; D014315:Triterpenes; C010480:lupeol; D000094042:Lupanes