EValuation of Azasterols as Anti-Parasitics
Journal of Medicinal Chemistry, 2006, Vol. 49, No. 20 6103
(6) Nes, W. D. Sterol methyl transferase: enzymology and inhibition.
BBA-Mol. Cell. Biol. Lipids 2000, 1529, 63-88.
(7) Venkatramesh, M.; Guo, D. A.; Jia, Z. H.; Nes, W. D. Mechanism
and structural requirements for transformation of substrates by the
(S)-adenosyl-L-methionine: Delta(24(25))- sterol methyl transferase
from Saccharomyces cereVisiae. BBA-Lipids Lipid Metab. 1996,
1299, 313-324.
(8) Ator, M. A.; Schmidt, S. J.; Adams, J. L.; Dolle, R. E.; Kruse, L. I.;
Frey, C. L.; Barone, J. M. Synthesis, specificity, and antifungal
activity of inhibitors of the Candida albicans D-24-sterol methyl-
transferase. J. Med. Chem. 1992, 35, 100-106.
(Schizotrypanum) cruzi epimastigotes grown in the presence of sterol
biosynthesis inhibitors. Biochem. Pharmacol. 1997, 53, 697-704.
(25) Vivas, J.; Urbina, J. A.; deSouza, W. Ultrastructural alterations in
Trypanosoma (Schizotrypanum) cruzi induced by Delta(24(25)) sterol
methyl transferase inhibitors and their combinations with ketocona-
zole. Int. J. Antimicrob. Agents 1996, 7, 235-240.
(26) Urbina, J. A.; Vivas, J.; Visbal, G.; Contreras, L. M. Modification
of the sterol composition of Trypanosoma (Schizotrypanum) cruzi
epimastigotes by delta(24(25))-sterol methyl transferase inhibitors
and their combinations with ketoconazole. Mol. Biochem. Parasitol.
1995, 73, 199-210.
(9) Ator, M. A.; Schmidt, S. J.; Adams, J. L.; Dolle, R. E. Mechanism
and inhibition of delta-24-sterol methyltransferase from Candida
albicans and Candida tropicalis. Biochemistry 1989, 28, 9633-9640.
(10) Oehlschlager, A. C.; Angus, R. H.; Pierce, A. M.; Pierce, H. D.;
Srinivasan, R. Azasterol inhibition of delta-24-sterol methyltransferase
in Saccharomyces cereVisiae. Biochemistry 1984, 23, 3582-3589.
(11) Janssen, G. G.; Nes, W. D. Structural requirements for transformation
of substrates by the S-adenosyl-L-methioninesdelta-24(25)-sterol
methyltransferasesinhibition by analogues of the transition-state
coordinate. J. Biol. Chem. 1992, 267, 25856-25863.
(12) Pierce, H. D.; Pierce, A. M.; Srinivasan, R.; Unrau, A. M.;
Oehlschlager, A. C. Azasterol inhibitors in yeast. Inhibition of the
24-methylene sterol D24(28)-reductase and D24-sterol methyltransferase
from Saccharomyces cereVisiae by 23-azacholesterol. Biochim.
Biophys. Acta 1978, 529, 429-437.
(13) Acuna, A. P.; Oehlschlager, A. C. Yeast delta-24-sterolmethyltrans-
ferase - inhibition by sulfonium analogues of the presumptive
cationic intermediates. Arch. Biol. Med. Exp. 1987, 20, R171-R171.
(14) Urbina, J. A.; Visbal, G.; Contreras, L. M.; McLaughlin, G.;
Docampo, R. Inhibitors of delta(24(25)) sterol methyltransferase block
sterol synthesis and cell proliferation in Pneumocystis carinii.
Antimicrob. Agents Chemother. 1997, 41, 1428-1432.
(27) Urbina, J. A.; Vivas, J.; Lazardi, K.; Molina, J.; Payares, G.; Piras,
M. M.; Piras, R. Antiproliferative effects of Delta 24(25) sterol methyl
transferase inhibitors on Trypanosoma (Schizotrypanum) cruzi: In
vitro and in vivo studies. Chemotherapy 1996, 42, 294-307.
(28) Rodrigues, J. C. F.; Attias, M.; Rodriguez, C.; Urbina, J. A.; de Souza,
W. Ultrastructural and biochemical alterations induced by 22,26-
azasterol, a Delta(24(25))-sterol methyltransferase inhibitor, on
promastigote and amastigote forms of Leishmania amazonensis.
Antimicrob. Agents Chemother. 2002, 46, 487-499.
(29) Haughan, P. A.; Chance, M. L.; Goad, L. J. Effects of an azasterol
inhibitor of sterol 24-transmethylation on sterol biosynthesis and
growth of Leishmania donoVani promastigotes. Biochem. J. 1995,
308, 31-38.
(30) Magaraci, F.; Jimenez, C. J.; Rodrigues, C.; Rodrigues, J. C. F.; Braga,
M. V.; Yardley, V.; de Luca-Fradley, K.; Croft, S. L.; de Souza,
W.; Ruiz-Perez, L. M.; Urbina, J.; Pacanowska, D. G.; Gilbert, I. H.
Azasterols as inhibitors of sterol 24-methyltransferase in leishmania
species and Trypanosoma cruzi. J. Med. Chem. 2003, 46, 4714-
4727.
(31) Coppens, I.; Courtoy, P. J. The adaptative mechanisms of Trypano-
soma brucei for sterol homeostasis in its different life-cycle environ-
ments. Annu. ReV. Microbiol. 2000, 54, 129-156.
(15) Rahier, A.; Genot, J. C.; Schuber, F.; Benveniste, P.; Narula, A. S.
Inhibition of S-adenosyl-L-methionine sterol-C-24-methyltransferase
by analogues of a carbocationic ion high-energy intermediate -
Structure activity relationships for C-25 heteroatoms (N, as, S)
substituted triterpenoid derivatives. J. Biol. Chem. 1984, 259, 5215-
5223.
(16) Narula, A. S.; Rahier, A.; Benveniste, P.; Schuber, F. 24-Methyl-
25-azacycloartanol, an analogue of a carbonium-ion high-energy
intermediate, is a potent inhibitor of (S)- adenosyl-L-methionine:
Sterol C-24-methyltransferase in higher-plant cells. J. Am. Chem.
Soc. 1981, 103, 2408-2409.
(17) Parks, L. W.; Smith, S. J.; Crowley, J. H. Biochemical and
physiological-effects of sterol alterations in yeast: A review. Lipids
1995, 30, 227-230.
(18) Nes, W. D.; McCourt, B. S.; Zhou, W. X.; Ma, J. Z.; Marshall, J.
A.; Peek, L. A.; Brennan, M. Overexpression, purification, and
stereochemical studies of the recombinant (S)-adenosyl-L-methion-
ine: Delta(24(25))- to Delta(24(28))-sterol methyl transferase enzyme
from Saccharomyces cereVisiae. Arch. Biochem. Biophys. 1998, 353,
297-311.
(19) Jia, Z. H.; Zhou, W.; Guo, D.; Nes, W. D. Synthesis of rationally
designed mechanism-based inactivators of the (S)-adenosyl-L-me-
thionine: Delta(24(25))-sterol methyl transferase. Synth. Commun.
1996, 26, 3841-3848.
(20) Lorente, S. O.; Jimenez, C. J.; Gros, L.; Yardley, V.; de Luca-Fradley,
K.; Croft, S. L.; Urbina, J. A.; Ruiz-Perez, L. M.; Pacanowska, D.
G.; Gilbert, I. H. Preparation of transition-state analogues of sterol
24-methyl transferase as potential anti-parasities. Bioorg. Med. Chem.
2005, 13, 5435-5453.
(21) Lorente, S. O.; Rodrigues, J. C. F.; Jimenez, C. J.; Joyce-Menekse,
M.; Rodrigues, C.; Croft, S. L.; Yardley, V.; de Luca-Fradley, K.;
Ruiz-Perez, L. M.; Urbina, J.; de Souza, W.; Pacanowska, D. G.;
Gilbert, I. H. Novel azasterols as potential agents for treatment of
leishmaniasis and trypanosomiasis. Antimicrob. Agents Chemother.
2004, 48, 2937-2950.
(22) Liendo, A.; Visbal, G.; Piras, M. M.; Piras, R.; Urbina, J. A. Sterol
composition and biosynthesis in Tryanosoma cruzi amastigotes. Mol.
Biochem. Parasitol. 1999, 104, 81-91.
(23) Vivas, J.; Urbina, J. A.; deSouza, W. Ultrastructural alterations in
Trypanosoma (Schizotrypanum) cruzi induced by Delta(24(25)) sterol
methyl transferase inhibitors and their combinations with ketocona-
zole. Int. J. Antimicrob. Agents 1997, 8, 1-6.
(24) Contreras, L. M.; Vivas, J.; Urbina, J. A. Altered lipid composition
and enzyme activities of plasma membranes from Trypanosoma
(32) Coppens, I.; Courtoy, P. J. Exogenous and endogenous sources of
sterols in the culture-adapted procyclic trypomastigotes of Trypano-
soma brucei. Mol. Biochem. Parasitol. 1995, 73, 179-188.
(33) Gros, L.; Castillo Acosta, V. M.; Jimenez Jimenez, C.; Sealy, M.;
Vargas Roldan, S.; Estevez, A.; Yardley, V.; Rattray, L.; Croft, S.
L.; Ruiz-Perez, L. M.; Urbina, J. A.; Gilbert, I. H.; Gonzalez
Pacanowska, D. Evaluation of new azasterols against Trypanosoma
brucei: Role of 24 sterol methyltransferase in inhibitor action.
Antimicrob. Agents Chemother., 2006, 50, 2595-2601.
(34) Vial, H. Isoprenoid biosynthesis and drug targeting in the aplicom-
plexa. Parasitol. Today 2000, 16, 140-141.
(35) Stock solutions of the test compounds plus control drugs were
prepared at a concentration of 20 mg/mL in DMSO (Sigma, U.K.).
The 2 times concentration 3-fold serial dilutions, in triplicate, of the
compounds were prepared in 96-well plates (Falcon, Life Technolo-
gies, U.K.).
(36) Neal, R. A.; Croft, S. L. An in vitro system for determining the
activity of compounds against the intracellular amastigote form of
Leishmania donoVani. J. Antimicrob. Chemother. 1984, 14, 463-
475.
(37) Buckner, F. S.; Verlinde, C. L. M. J.; La Flamme, A. C.; Van Voorhis,
W. C. Efficient technique for screening drugs for activity against
Trypanosoma cruzi parasites expressing beta-galactosidase. Antimi-
crob. Agents. Chemother. 1996, 40, 2592-2597.
(38) Hirumi, H.; Hirumi, K. Continuous cultivation of Trypanosoma brucei
blood stream forms in a medium containing a low concentration of
serum-protein without feeder cell-layers. J. Parasitol. 1989, 75, 985-
989.
(39) Raz, B.; Iten, M.; Grether-Buhler, Y.; Kaminski, R.; Brun, R. The
Alamar blue assay to determine drug sensitivity of African trypa-
nosomes (T. b. rhodesiense and T. b. gambiense) in vitro. Acta Trop.
1997, 68, 139-147.
(40) Desjardins, R. E.; Canfield, C. J.; Haynes, J. D.; Chulay, J. D.
Quantitative assesment of antimalarial activity in vitro by a semi-
automated microdilution technique. Antimicob. Agents Chemother.
1979, 16, 710-718.
(41) O’Neill, M. J.; Bray, D. H.; Boardman, P.; Phillipson, J. D.; Warhurst,
D. C. In Vitro test method for the evaluation of crude extracts from
plants. Planta Medica 1985, 61, 394-397.
(42) Anderson, M. M.; O’Neill, M. J.; Phillipson, J. D.; Warhurst, D. C.
In Vitro cytotoxicity of a series of quassinoids from Brucea jaVanica
fruits against KB cells. Planta Medica 1991, 57, 62-64.
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