annonacin and thiazolyl-blue

The acetogenin, annonacin, from the tropical annonaceous plant Annona muricata, is a lipophilic, mitochondrial complex I inhibitor reported to be more toxic than rotenone to mesencephalic neurons. The temperate annonaceous plant Asimina triloba (pawpaw) is native to the Eastern United States and products are available online. This study determined whether annonacin is in the pawpaw fruit pulp and whether it or the crude ethyl acetate extract is toxic to cortical neurons.. Pawpaw extract was prepared by pulp extraction with methanol and liquid-liquid partitioning with ethyl acetate (EtOAc). Annonacin was isolated from the crude EtOAc extract via column chromatography using a gradient solvent system of increasing polarity. Mass spectroscopy, nuclear magnetic resonance and infrared spectroscopy were used to compare isolated material with synthetic annonacin data and a natural annonacin sample. Toxicity of isolated annonacin and the total EtOAc extract was determined in primary rat cortical neurons using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay.. The average concentration of annonacin in the fruit pulp was 0.0701±0.0305mg/g. Purified annonacin (30.07μg/ml) and crude EtOAc extract (47.96μg/ml) induced 50% death of cortical neurons 48h post treatment. Annonacin toxicity was enhanced in the presence of crude extract.. Pawpaw fruit contains a high concentration of annonacin, which is toxic to cortical neurons. Crude fruit extract also induced neurotoxicity, highlighting the need for additional studies to determine the potential risks of neurodegeneration associated with chronic exposure to pawpaw products.

Topics: Animals; Animals, Newborn; Asimina; Cell Count; Cells, Cultured; Cerebral Cortex; Dose-Response Relationship, Drug; Fruit; Furans; Lactones; Magnetic Resonance Spectroscopy; Mass Spectrometry; Neurons; Neurotoxins; Rats; Rats, Sprague-Dawley; Spectrophotometry, Infrared; Tetrazolium Salts; Thiazoles

In this study we evaluated a mono-tetrahydrofuranic subgroup of natural acetogenins that had shown in previous enzyme inhibition studies different potency trends compared with the bis-tetrahydrofuranic acetogenin subgroup. The compounds were tested against colon, breast, lung, liver, and ovarian tumor cell lines. A drug-resistant ovarian cell line was also included in the panel. In general the compounds were more potent than doxorubicin. The goal was to determine how well the mitochondrial complex I inhibition correlates with the in vitro antitumor potency of these natural mono-tetrahydrofuranic acetogenins and of some derivatives. The results indicate that both the reduction of the terminal gamma-lactone after its translactonization and the introduction of an hydroxylimine group in the alkyl chain, near the mono-tetrahydrofuranic moiety, increased the antitumor activity, even against the doxorubicin-resistant cell line.

Topics: Acetogenins; Antineoplastic Agents; Cell Line, Tumor; Electron Transport Complex I; Fatty Alcohols; Furans; Humans; Inhibitory Concentration 50; Lactones; Molecular Structure; Multienzyme Complexes; NADH, NADPH Oxidoreductases; Structure-Activity Relationship; Tetrazolium Salts; Thiazoles