Ginkgolide B and bilobalide block the pore of the 5-HT3 receptor at a location that overlaps the picrotoxin binding site

Extracts from the Ginkgo biloba tree are widely used as herbal medicines, and include bilobalide (BB) and ginkgolides A and B (GA and GB). Here we examine their effects on human 5-HT3A and 5-HT3AB receptors, and compare these to the effects of the structurally related compounds picrotin (PTN) and picrotoxinin (PXN), the two components of picrotoxin (PTX), a known channel blocker of 5-HT3, nACh and GABAA receptors. The compounds inhibited 5-HT-induced responses of 5-HT3 receptors expressed in Xenopus oocytes, with IC50 values of 470μM (BB), 730μM (GB), 470μM (PTN), 11μM (PXN) and >1mM (GA) in 5-HT3A receptors, and 3.1mM (BB), 3.9mM (GB), 2.7mM (PTN), 62 μM (PXN) and >1mM (GA) in 5-HT3AB receptors. Radioligand binding on receptors expressed in HEK 293 cells showed none of the compounds displaced the specific 5-HT3 receptor antagonist [3H]granisetron, confirming that they do not act at the agonist binding site. Inhibition by GB at 5-HT3A receptors is weakly use-dependent, and recovery is activity dependent, indicating channel block. To further probe their site of action at 5-HT3A receptors, BB and GB were applied alone or in combination with PXN, and the results fitted to a mathematical model; the data revealed partially overlapping sites of action. We conclude that BB and GB block the channel of the 5-HT3A receptor. Thus these compounds have comparable, although less potent, behaviour than at some other Cys-loop receptors, demonstrating their actions are conserved across the family. ► Ginkgolide B and bilobalide inhibit 5-HT3 receptors. ► They are more potent at 5-HT3A than 5-HT3AB receptors. ► BB and GB block the open channel of 5-HT3A receptors. ► Their sites of action partially overlap with those of the channel blocker picrotoxinin.