Zebrafish-Based Screening of Antiseizure Plants Used in Traditional Chinese Medicine: Magnolia officinalis Extract and Its Constituents Magnolol and Honokiol Exhibit Potent Anticonvulsant Activity in a Therapy-Resistant Epilepsy Model

With the aim to discover interesting lead compounds that could be further developed into compounds active against pharmacoresistant epilepsies, we first collected 14 medicinal plants used in traditional Chinese medicine (TCM) against epilepsy. Of the six extracts that tested positive in a pentylenetetrazole (PTZ) behavioral zebrafish model, only the ethanol and acetone extracts from Magnolia officinalis (M. officinalis) also showed effective antiseizure activity in the ethylketopentenoate (EKP) zebrafish model. The EKP model is regarded as an interesting discovery platform to find mechanistically novel antiseizure drugs, as it responds poorly to a large number of marketed anti-epileptics. We then demonstrated that magnolol and honokiol, two major constituents of M. officinalis, displayed an effective behavioral and electrophysiological antiseizure activity in both the PTZ and the EKP models. Out of six structural analogues tested, only 4-O-methylhonokiol was active and to a lesser extent tetrahydromagnolol, whereas the other analogues (3,3′-dimethylbiphenyl, 2,2′-biphenol, 2-phenylphenol, and 3,3′,5,5′-tetra-tert-butyl-[1,1′-biphenyl]-2,2′-diol) were not consistently active in the aforementioned assays. Finally, magnolol was also active in the 6 Hz psychomotor mouse model, an acute therapy-resistant rodent model, thereby confirming the translation of the findings from zebrafish larvae to mice in the field of epilepsy. We also developed a fast and automated power spectral density (PSD) analysis of local field potential (LFP) recordings. The PSD results are in agreement with the visual analysis of LFP recordings using Clampfit software and manually counting the epileptiform events. Taken together, screening extracts of single plants employed in TCM, using a combination of zebrafish- and mouse-based assays, allowed us to identify allyl biphenol as a chemical scaffold for the future development of compounds with potential activity against therapy-resistant epilepsies.