Inhibitors of delta24(25) sterol methyltransferase block sterol synthesis and cell proliferation in Pneumocystis carinii.
Detailed analysis of the endogenous sterol content of purified Pneumocystis carinii preparations by gas-liquid chromatography coupled to mass spectrometry suggested that this parasite can both synthesize de novo steroid skeletons (to produce delta7 sterols) and take them from the infected host (leading to delta5 sterols). In both cases the final products are 24-alkyl sterols, resulting from the action of delta24(25) and delta24(24') sterol methyltransferases, enzymes not present in vertebrates. To investigate the physiological significance of these sterols, cultures of P. carinii in embryonic lung cells were exposed to 22,26-azasterol (20-piperidin-2-yl-5alpha-pregnan-3beta-20(R)-diol), a compound previously shown to inhibit both enzymes and to halt cell proliferation in fungi and protozoa. This compound produced a dose-dependent reduction in the parasite proliferation, with a 50% inhibitory concentration of 0.3 microM and 80% reduction of growth after 96 h at 10 microM. Correspondingly, parasites treated with the azasterol at 10 microM for 48 h accumulated 24-desalkyl sterols such as zymosterol (cholesta-8,24-dien-3beta-ol) and cholesta-8,14,24-trien-3beta-ol to ca. 40% of the total mass of endogenous sterols. This is the first report on the antiproliferative effects of a sterol biosynthesis inhibitor on P. carinii and indicate that sterol methyltransferase inhibitors could be the basis of a novel and specific chemotherapeutic approach to the treatment of P. carinii infections.
Urbina
J A
JA
Laboratorio de Química Biológica, Centro de Biofísica y Bioquímica,Instituto Venezolano de Investigaciones Científicas, Caracas, Venezuela. jaurbina@churchill.scs.uiuc.edu
Visbal
G
G
Contreras
L M
LM
McLaughlin
G
G
Docampo
R
R
D000478:Alkylation; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D004083:Cholestanol / Q000031:analogs & derivatives / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D008168:Lung / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008780:Methyltransferases / Q000037:antagonists & inhibitors*; D011010:Pneumocystis / Q000187:drug effects* / Q000378:metabolism; D013261:Sterols / Q000096:biosynthesis*
C103731:22,26-azasterol; D004791:Enzyme Inhibitors; D013261:Sterols; D004083:Cholestanol; D008780:Methyltransferases; C022670:delta 24-sterol methyltransferase