Total Structure Determination of Apratoxin A, a Potent Novel Cytotoxin from the Marine Cyanobacterium Lyngbya majuscula

• Published in: Journal of the American Chemical Society 2001-06, Vol.123 (23), p.5418-5423
• Main Authors: LUESCH, Hendrik, YOSHIDA, Wesley Y., MOORE, Richard E., PAUL, Valerie J., CORBETT, Thomas H.
• Format: Article
• Language: English
• Subjects: Adenocarcinoma - drug therapy; Animals; Bacterial Toxins - chemistry; Bacterial Toxins - isolation & purification; Bacterial Toxins - pharmacology; Breast Neoplasms - drug therapy; Cell Survival - drug effects; Colonic Neoplasms - drug therapy; Cyanobacteria - chemistry; Cytotoxins - chemistry; Cytotoxins - isolation & purification; Cytotoxins - pharmacology; Depsipeptides; Female; Humans; Inhibitory Concentration 50; Male; Marine Toxins - chemistry; Marine Toxins - isolation & purification; Marine Toxins - pharmacology; Mice; Molecular Structure; Nuclear Magnetic Resonance, Biomolecular; Peptides, Cyclic - chemistry; Peptides, Cyclic - isolation & purification; Peptides, Cyclic - pharmacology; Tumor Cells, Cultured
• ISSN: 0002-7863; 1520-5126
• DOI: 10.1021/ja010453j

Apratoxin A (1), a potent cytotoxin with a novel skeleton, has been isolated from the marine cyanobacterium Lyngbya majuscula Harvey ex Gomont. This cyclodepsipeptide of mixed peptide-polyketide biogenesis bears a thiazoline ring flanked by polyketide portions, one of which possesses an unusual methylation pattern. Its gross structure has been elucidated by spectral analysis, including various 2D NMR techniques. The absolute configurations of the amino acid-derived units were determined by chiral HPLC analysis of hydrolysis products. The relative stereochemistry of the new dihydroxylated fatty acid unit, 3,7-dihydroxy-2,5,8,8-tetramethylnonanoic acid, was elucidated by successful application of the J-based configuration analysis originally developed for acyclic organic compounds using carbon-proton spin-coupling constants ((2,3)J(C,H)) and proton-proton spin-coupling constants ((3)J(H,H)); its absolute stereochemistry was established by Mosher analysis. The conformation of 1 in solution was mimicked by molecular modeling, employing a combination of distance geometry and restrained molecular dynamics. Apratoxin A (1) possesses IC(50) values for in vitro cytotoxicity against human tumor cell lines ranging from 0.36 to 0.52 nM; however, it was only marginally active in vivo against a colon tumor and ineffective against a mammary tumor.