15-methoxypinusolidic-acid and pinusolide

Apoptosis is a fundamental process required for neuronal development but also occurs in most of the common neurodegenerative disorders. In an attempt to obtain an anti-apoptotic neuroprotective compound from natural products, we isolated the diterpenoids, pinusolide and 15-MPA, from B. orientalis and investigated their neuroprotective activity against staurosporine (STS) -induced neuronal apoptosis. In addition, we determined the anti-apoptotic mechanism of these compounds in rat cortical cells.. Primary cultures of rat cortical cells injured by STS were used as an in vitro assay system. Cells were pretreated with pinusolide or 15-MPA before exposure to STS. Anti-apoptotic activities were evaluated by the measurement of cytoplasmic condensation and nuclear fragmentation. The levels of cellular peroxide, malondialdehyde (MDA) and [Ca(2+)]i, as well as the activities of superoxide dismutase (SOD) and caspase-3/7, were measured.. Pinusolide and 15-MPA, at a concentration of 5.0 ìM, reduced the condensed nuclei and rise in [Ca(2+)]i that accompanies apoptosis induced by 100 nM STS. Pinusolide and 15-MPA also protected the cellular activity of SOD, an antioxidative enzyme reduced by STS insult. Furthermore, the overproduction of reactive oxygen species and lipid peroxidation induced by STS was significantly reduced in pinusolide and 15-MPA treated cells. In addition, pinusolide and 15-MPA inhibited STS-induced caspase-3/7 activation.. These results show that pinusolide and 15-MPA protect neuronal cells from STS-induced apoptosis, probably by preventing the increase in [Ca(2+)]i and cellular oxidation caused by STS, and indicate that they could be used to treat neurodegenerative diseases.

Topics: Animals; Calcium; Caspases; Cells, Cultured; Cerebral Cortex; Diterpenes; Enzyme Activation; Malondialdehyde; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Staurosporine; Superoxide Dismutase

A new pinusolide derivative, 15-methoxypinusolidic acid (1), and another new isopimarane diterpene, ent-isopimara-15-en-3 alpha,8 alpha-diol (2) with three known diterpenes, lambertianic acid (3), isopimara-8(9),15-dien-18-oic acid (4) and isopimara-7(8),15-dien-3 beta,18-diol (5) were isolated from the 90% MeOH fraction of Biota orientalis (L.) ENDL. (Cupressaceae) leaves. Chemical structures of 1-5 were elucidated by analyses of their spectral data, including the two-dimensional (2D) NMR technique. Compound 1 showed significant protective activity against glutamate-induced neurotoxicity in primary cultures of rat cortical cells.

Topics: Biodiversity; Diterpenes; Drugs, Chinese Herbal; Neuroprotective Agents; Plant Leaves; Thuja