School of Natural Product Studies, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, 700 032, India.
School of Natural Product Studies, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, 700 032, India; Institute of Bioresources and Sustainable Development, An Autonomous Institute Under Dept. of Biotechnology, Govt. of India, Imphal, 795001, India.
J Ethnopharmacol. 2021 Feb 10;266:113457. doi: 10.1016/j.jep.2020.113457. Epub 2020 Oct 9.
Trimada is well-known polyherbal Ayurvedic formulation used in Indian Traditional medicine since ancient times. It consisted of three inebriant herbs including "Chitraka" (Plumbago zeylanica Linn. Family- Plumabaginaceae), "Musta" (Cyperus rotundus Linn. Family- Cyperaceae) and Vidanga (Embelia ribes Burm. F. Family- Myrsinaceae) in equal ratios as mentioned in Ayurveda. Trimada is traditionally used to increase the functioning of the digestive system and metabolism. Along with these, it also assists in the reduction of cholesterol as well as reduces stomach aches and chest pain.
This study is aimed to identify the metabolites present in this polyherbal formulation. Further, the cytotoxicity and interaction potential of the formulation and individual herbs with Cytochrome P450 isozymes (CYP3A4, 2D6, 2C9, 1A2) was evaluated by MTT assay and CYP450 enzyme inhibition. The concentration of heavy metals was also determined.
Ultra-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry (UPLC-QTOF-MS) analysis was performed to detect and identify the phytoconstituents in the formulation. Cytotoxicity of the formulation was evaluated by MTT assay. CYP450 enzyme interaction potential of the individual herbs and the Trimada formulation was carried out through CYP-CO assay and fluorometric high throughput screening (HTS) assay for individual isozymes. The content of heavy metal in the formulation was quantified by Atomic Absorption Spectroscopy.
Trimada formulation exhibited lower cytotoxicity to human liver carcinoma cell line (HepG2). CYP-CO assay revealed that the interaction potential of individual herbs and Trimada on the liver microsomes was found to be lesser than the standard inhibitor ketoconazole. Individual herbs and Trimada formulation displayed higher IC values than the respective standard inhibitors in the fluorimetric assay. UPLC-QTOF-MS analysis showed the presence of a number of active phytoconstituents including sesquiterpenes, phenolic acids, benzoquinones, triterpenes and flavonoids. The heavy metal concentration in the traditional medicinal herbal formulation was found within the approved limit.
This study suggested that the individual herbs and Trimada formulation exhibited low cytotoxicity and contributes insignificant interaction with CYP450 isozymes. So, the formulation is considered to be safe for its therapeutic management without any potential drug interaction involving CYP 450 isozymes.
Trimada 是一种广为人知的复方印度草药制剂，自古以来就在印度传统医学中使用。它由三种致醉草药组成，包括阿育吠陀中提到的“Chitraka”（Plumbago zeylanica Linn. 科- Plumabaginaceae）、“Musta”（Cyperus rotundus Linn. 科- Cyperaceae）和 Vidanga（Embelia ribes Burm. F. 科- Myrsinaceae），比例相等。Trimada 传统上用于增强消化系统和新陈代谢的功能。除此之外，它还可以帮助降低胆固醇，减少胃痛和胸痛。
本研究旨在鉴定这种复方制剂中的代谢产物。此外，通过 MTT 测定和 CYP450 酶抑制测定，评估了该制剂和各草药与细胞色素 P450 同工酶（CYP3A4、2D6、2C9、1A2）的细胞毒性和相互作用潜力。还测定了重金属的浓度。
采用超高效液相色谱-四极杆/飞行时间质谱（UPLC-QTOF-MS）分析检测和鉴定制剂中的植物成分。通过 MTT 测定评估制剂的细胞毒性。通过 CYP-CO 测定和荧光高通量筛选（HTS）测定各同工酶，研究各草药和 Trimada 制剂对 CYP450 酶的相互作用潜力。采用原子吸收光谱法测定制剂中重金属的含量。
Trimada 制剂对人肝癌细胞系（HepG2）的细胞毒性较低。CYP-CO 测定显示，各草药和 Trimada 对肝微粒体的相互作用潜力小于标准抑制剂酮康唑。在荧光测定中，各草药和 Trimada 制剂的 IC 值均高于各自的标准抑制剂。UPLC-QTOF-MS 分析表明，存在多种活性植物成分，包括倍半萜、酚酸、苯醌、三萜和类黄酮。传统药用草药制剂中的重金属浓度在批准范围内。
本研究表明，各草药和 Trimada 制剂的细胞毒性较低，与 CYP450 同工酶的相互作用不显著。因此，该制剂在治疗管理中被认为是安全的，不会因涉及 CYP450 同工酶的潜在药物相互作用而产生任何问题。
