ES 242-1

ES-242-1, a novel bioxanthracene of microbial origin, blocked glutamate-induced neuronal death in a dose-dependent manner at concentrations ranging from 0.01 to 1.0 microM, but not the neuronal death caused by kainic acid or quisqualic acid. ES-242-1 also prevented cell death induced by 2,4-methanoglutamate, which is a specific agonist for the NMDA receptor. ES-242-1 showed protective effects in cultured neurons prepared from cerebellum and septum as it did in cultured hippocampal neurons but to different extents. There was a positive correlation between the potencies of ES-242s as inhibitors of ligand binding to the NMDA receptor and as inhibitors of neuronal death. Hypoxic treatment for 4 h under 95% N2 and 5% CO2 caused neuronal death of the cultured hippocampal neurons. Again, ES-242-1 at 1.0 microM was effective to protect neurons against hypoxic injury. ES-242 compounds are new chemical entities possessing neuroprotective properties useful in the treatment of diseases involving glutamate toxicity.

The effects of a novel N-methyl-D-aspartate (NMDA) receptor antagonist, ES-242-1 (3,3'-dimethyl-3,4,3',4'-tetrahydro-6,8,6',8'-tetramethoxy-[10,10' - bi-2-oxanthracene]-4,9,9'-(1H,1'H)-triol 4-acetate), on NMDA-induced increases of intracellular Ca2+ concentration in cultured hippocampal neurons were examined. ES-242-1 selectively blocked the NMDA-induced increase in intracellular free Ca2+ concentration ([Ca2+]i), but not the [Ca2+]i increase stimulated by quisqualate or kainate. The effect of ES-242-1 appeared in the slow development of a blockade of [Ca2+]i (half blocking time: 90 sec) when 100 microM NMDA was applied with 10 microM ES-242-1, whereas the initial [Ca2+]i rise was attenuated by 10 microM ES-242-1 when the latter was applied with a lower concentration of NMDA (10 microM). This is consistent with a previous observation that ES-242-1 binds to both the transmitter recognition site and the channel domain. The blockade by ES-242-1 was reversed by washing. In contrast, the blockade by MK-801 was not relieved easily by washing. These results suggest that ES-242-1 blocks the NMDA-induced [Ca2+]i increase due to a combination of two well-recognized mechanisms, which are different from that of MK-801, at the NMDA receptor.

The purpose of this study was to determine whether ES-242-1, a novel N-methyl-D-aspartate (NMDA) receptor antagonist of microbial origin, has anti-nociception at the spinal level and to evaluate how its anti-nociceptive effect differs from that of MK-801, a non-competitive NMDA receptor antagonist. Agents were injected intrathecally (0.1, 1.0 and 10 microg) through a previously implanted PE tube in rats. Formalin (2%, 100 microl) was injected subcutaneously into the left hindpaw 15 min after each antagonist administration. Licking time as a nociceptive behavior was measured in three stages after formalin-injection, such as early phase (0-9 min), late first phase (10-29 min) and late second phase (30-60 min). In the early phase, the largest dose of ES-242-1 significantly decreased total licking time, although MK-801 did not show any significant reduction. With the treatment of 1.0 and 10 microg MK-801, total licking time in both late first and second phases was significantly suppressed, although the smallest dose (0.1 microg) of ES-242-1 showed a significant reduction in the late second phase. These results indicate that ES-242-1 is highly effective against tonic pain, such as inflammatory pain.