Nóra Gampe 1, László Kursinszki 1, Szabolcs Béni 1
1 Semmelweis University, Department of Pharmacognosy, Budapest
Introduction: Based on our previous results, Ononis spinosa L. contains a wide variety of isoflavonoids, of which formononetin showed favorable results in the treatment of Alzheimer-disease in vivo and in vitro, maackiain could inhibit selectively the MAO-B enzyme. Regarding these outcomes, Ononis species could be rich sources of compounds affecting the CNS.
Aims: Firstly, we aimed to investigate the qualitative composition of O. arvensis and compare its constituents with that of O. spinosa. Furthermore, quantitative analytical methods were needed for the exploration of the isoflavonoid content. Since with biotechnology secondary metabolite production can be multiplied, the investigation of the isoflavonoid production of in vitro cultures was aimed. Moreover, we intended to study the CNS permeability these isoflavonoids.
Methods: The main technique of the qualitative investigation was UHPLC-HR-MS/MS supplemented with NMR experiments. For the quantitative measurements, HPLC-DAD and HPLC-ESI-MS/MS methods were developed. The isolation of the compounds was executed using Flash-chromatography and preparative HPLC. The permeability through the blood-brain barrier was estimated by PAMPA-BBB model.
Results: During the qualitative investigation of O. arvensis significant differences in the isoflavonoid spectrum of the aerial parts and the roots were found. Based on the results of the quantitative analysis both Ononis species are very rich in isoflavonoids, but the hairy root cultures of O. spinosa exceeded all samples with outstandingly high total-isoflavonoid content. However, we found remarkable differences between the isoflavonoid profile of the in vitro and the free-range samples. According to the PAMPA-BBB measurements, all investigated isoflavonoid could pass the brain-blood barrier.
Conclusions: Ononis species, and especially their in vitro cultures, are rich sources of isoflavonoids with potential CNS effects. As these compounds can pass the brain-blood barrier, they are promising target molecules for further pharmacological tests.
Supported by the ÚNKP-18-3-III-SE-30 New National Excellence Program of the Ministry of Human Capacities.
Doctoral School: Doctoral School of Pharmaceutical Sciences
Program: Modern Trends in Pharmaceutical Scientific Research
Supervisors: Szabolcs Béni, László Kursinszki
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