Identification of Cubebin from Cubeb (Piper cubeba) as An Antiplatelet Agent

Platelets play a key role in arterial thrombosis. Platelet aggregation is part of a sequential mechanism leading to the formation of a thrombus. Antiplatelet agent is needed for the inhibition of platelet aggregation. Piper cubeba or Cubeb is an Indonesian traditional herbal medicine. Cubeb fruit is rich in lignans, such as cubebin, hinokinin, clusin, dihydrocubebin, dihydroclusin, cubebinin, yatein, cubbinulide (cordigerine), dihydroyatein, isoyatein, and cubebinone. Previous studies have indicated that several lignans exert antiplatelet activity. Additionally, the extract of Piper cubeba fruit exhibits antiplatelet activity. This study aimed to evaluate the antiplatelet activities of P. cubeba fruit fractions and identify active compound. The fruit was macerated in ethanol for 5 days. The extract was fractionated by vacuum liquid chromatography (using dichloromethane with ethylacetate added in a polar gradient. The main compound was isolated and identified based on its infrared, H-nuclear magnetic resonance, (H-NMR), C-NMR, and mass spectra. The antiplatelet activity was determined by an in vitro antiplatelet aggregation assay employing human platelets induced with several platelet receptor agonists Adenosine Diphosphate (ADP), epinephrine, thrombin, and arachidonic acid). The ethanol extract of P. cubeba fruit and its fractions demonstrated antiplatelet activity induced by epinephrine, ADP, thrombin, arachidonic acid and ristocetin. The major compound in the active fraction was identified as cubebin. Cubebin demonstrated antiplatelet activity in platelet aggregation induced by thrombin with the IC50 of 14.62±1.16 µM. Cubebin is a novel antiplatelet agent isolated from P. cubeba. © 2023 Herlyanti et al.