Harwansh Ranjit K, Mukherjee Kakali, Bhadra Santanu, Kar Amit, Bahadur Shiv, Mitra Achintya, Mukherjee Pulok K
School of Natural Product Studies, Department of Pharmaceutical Technology, Jadavpur University, Kolkata 700032, India.
National Research Institute of Ayurvedic Drug Development, Central Council for Research in Ayurvedic Sciences, Department of AYUSH, Kolkata 700091, India.
J Ethnopharmacol. 2014 May 14;153(3):674-81. doi: 10.1016/j.jep.2014.03.023. Epub 2014 Mar 29.
Trikatu is a very well known 'Rasayana' in Ayurveda and widely used as a polyherbal ayurvedic formulation in India. It consists of three well known plants, viz., Piper longum (PL), Piper nigrum (PN) and Zingiber officinale (ZO) in equal ratio. Trikatu has been prescribed for cough, cold, fever, asthma, respiratory problems and improvement of digestive disorders. The aim of the present study was to investigate the effect of individual ingredients of trikatu namely PL, PN, and ZO and formulations [Marketed formulation (MF) and laboratory formulation (LF)] on drug metabolizing enzymes (CYP3A4 and CYP2D6), to assess its herb-drug interaction potential through cytochrome P450 inhibition assays. Further this work was aimed to develop an RP-HPLC method for the identification and quantification of piperine and 6-gingerol in the crude drug trikatu.
Enzyme inhibition effect of LF, MF, PL, PN and ZO was explored through CYP450-CO complex assay using rat liver microsomes (RLM) and a fluorescence screening method using individual isoenzymes (CYP3A4 and CYP2D6). The RP-HPLC method was developed for the identification and quantification of piperine and 6-gingerol in LF, MF and individual plant materials at the concentration of 1mg/mL.
RP-HPLC analysis confirmed the presence of piperine and 6-gingerol in LF and MF [Piperine: 7.89±2.12% (w/w) (MF), 6.70±2.13% (w/w) (LF)]; [6-gingerol: 5.3±1.21% (w/w) (MF), 4.95±2.34% (w/w) (LF)]. Inhibitory potential of MF and LF in CYP450-CO complex assay was found to be 37.54±3.12% (MF) and 35.12±2.31% (LF) and against CYP2D6 and CYP3A4 was estimated to be IC50 251.30±3.98 and 245.23±1.92μg/mL and IC50 225.50±1.02 and 223.254±0.92μg/mL respectively.
Different concentrations of the trikatu formulation and its individual components showed significantly (p<0.001) less inhibitory activity on individual isoenzymes as compared to the positive control. The crude drug exhibited inhibitory potential against the CYP450 enzymes in a concentration dependent manner. Outcome of the present study demonstrated that trikatu has less interaction potential with drug metabolizing enzymes.
三果木是阿育吠陀中一种非常著名的“rasayana”，在印度作为一种多草药阿育吠陀配方被广泛使用。它由三种著名植物组成，即长胡椒（PL）、黑胡椒（PN）和姜（ZO），比例相等。三果木已被用于治疗咳嗽、感冒、发烧、哮喘、呼吸道问题以及改善消化系统疾病。本研究的目的是研究三果木的各个成分，即PL、PN和ZO以及配方[市售配方（MF）和实验室配方（LF）]对药物代谢酶（CYP3A4和CYP2D6）的影响，通过细胞色素P450抑制试验评估其草药 - 药物相互作用潜力。此外，这项工作旨在开发一种反相高效液相色谱法（RP - HPLC），用于鉴定和定量粗药三果木中的胡椒碱和6 - 姜酚。
通过使用大鼠肝微粒体（RLM）的CYP450 - CO复合物测定法以及使用个体同工酶（CYP3A4和CYP2D6）的荧光筛选方法，探索LF、MF、PL、PN和ZO的酶抑制作用。开发了RP - HPLC方法，用于鉴定和定量LF、MF和个体植物材料中浓度为1mg/mL的胡椒碱和6 - 姜酚。
RP - HPLC分析证实LF和MF中存在胡椒碱和6 - 姜酚[胡椒碱：7.89±2.12%（w/w）（MF），6.70±2.13%（w/w）（LF）]；[6 - 姜酚：5.3±1.21%（w/w）（MF），4.95±2.34%（w/w）（LF）]。在CYP450 - CO复合物测定中，MF和LF的抑制潜力分别为37.54±3.12%（MF）和35.12±2.31%（LF），对CYP2D6和CYP3A4的IC50估计分别为251. / 30±3.98和245.23±1.92μg/mL以及IC50 225.50±1.02和223.254±0.92μg/mL。
与阳性对照相比，不同浓度的三果木配方及其各个成分对个体同工酶的抑制活性显著降低（p<0.001）。粗药对CYP450酶表现出浓度依赖性的抑制潜力。本研究结果表明，三果木与药物代谢酶的相互作用潜力较小。
