Zayed Aref L, Hadieh Mohammad, Al Hroot Jomana, Hameedat Fatima, Jaber Sana'a A
Department of Medicinal Chemistry and Pharmacognosy, Faculty of Pharmacy, Jordan University of Science and Technology, P.O. Box 3030, Irbid, 22110, Jordan.
Heliyon. 2024 May 14;10(10):e31266. doi: 10.1016/j.heliyon.2024.e31266. eCollection 2024 May 30.
Warfarin, a widely prescribed anticoagulant, is highly effective for various coagulation disorders. However, its efficacy is limited by a narrow therapeutic index and frequent drug interactions, especially those involving metabolism by Cytochrome P450 (CYP450) enzymes. Piperine, found in black and long pepper, possesses blood-thinning properties and has been observed to inhibit CYP3A and CYP2C enzymes linked to warfarin metabolism. This study investigated the effect of piperine on warfarin metabolism in liver microsomes using a rapid and sensitive HPLC-Fluorescence method. The use of PFP (pentafluorophenyl) column with core shell particles provided the selectivity and resolution to resolve warfarin and its 4-, 6-, 7-, and 10-hydroxy metabolites in addition to the internal standard naproxen in less than 3 min. This is the fastest analytical assay for warfarin and its major metabolites reported to date, making it ideal for metabolic studies. The applicability of the method was demonstrated by monitoring the metabolism of S-warfarin in human and rat liver microsomes, and evaluating the inhibitory effect of piperine on metabolite formation. The results showed that piperine inhibited the formation of the major metabolite, 7-hydroxywarfarin, with half-maximal inhibitory concentration (IC) 14.2 μM and 3.2 μM in human and rat liver microsomes, respectively. Furthermore, coagulation studies using rat plasma showed that piperine does not affect prothrombin time (PT) and activated partial thromboplastin time (aPTT). This study suggested that piperine may present a potential drug interaction with warfarin at the metabolism level, but has no direct effect on the activation of the extrinsic or intrinsic coagulation cascades. Further clinical investigation is therefore required, as piperine may increase the bioavailability of warfarin, thus increasing risk of serious adverse events in patients.
华法林是一种广泛使用的抗凝剂，对各种凝血障碍都非常有效。然而，其疗效受到狭窄治疗指数和频繁药物相互作用的限制，尤其是那些涉及细胞色素P450（CYP450）酶代谢的相互作用。胡椒碱存在于黑胡椒和长胡椒中，具有血液稀释特性，并且已观察到它能抑制与华法林代谢相关的CYP3A和CYP2C酶。本研究使用快速灵敏的高效液相色谱 - 荧光法研究了胡椒碱对肝微粒体中华法林代谢的影响。使用带有核壳颗粒的五氟苯基（PFP）柱，除内标萘普生外，能在不到3分钟的时间内分离华法林及其4 -、6 -、7 - 和10 - 羟基代谢物，提供了选择性和分辨率。这是迄今为止报道的对华法林及其主要代谢物最快的分析方法，使其成为代谢研究的理想方法。通过监测人及大鼠肝微粒体中S - 华法林的代谢，并评估胡椒碱对代谢物形成的抑制作用，证明了该方法具有适用性。结果表明，胡椒碱抑制主要代谢物7 - 羟基华法林的形成，在人及大鼠肝微粒体中的半数最大抑制浓度（IC）分别为14.2 μM和3.2 μM。此外，使用大鼠血浆进行的凝血研究表明，胡椒碱不影响凝血酶原时间（PT）和活化部分凝血活酶时间（aPTT）。本研究表明，胡椒碱可能在代谢水平上与华法林存在潜在的药物相互作用，但对外部或内源性凝血级联反应的激活没有直接影响。因此，由于胡椒碱可能增加华法林的生物利用度，从而增加患者发生严重不良事件的风险，所以需要进一步的临床研究。
