Department of Pharmaceutical Biology, Institute of Pharmaceutical and Biomedical Sciences, Johannes Gutenberg University, Staudinger Weg 5, 55128, Mainz, Germany; Department of Biochemistry, Faculty of Science, University of Dschang, Dschang, Cameroon.
Department of Biochemistry, Faculty of Science, University of Dschang, Dschang, Cameroon.
J Ethnopharmacol. 2021 May 10;271:113884. doi: 10.1016/j.jep.2021.113884. Epub 2021 Jan 30.
Piper capense is a medicinal spice whose fruits are traditionally used as aqueous decoction to heal several ailments such as trypanosomiasis, helminthic infections, and cancer.
(1) To perform phytochemical investigation of the methanol extract of Piper capense; (2) to evaluate the cytotoxicity of botanicals (PCF, fractions PCFa-e), isolated phytochemicals on a broad panel of animal and human cancer cell lines; (3) to evaluate the induction of apoptosis of the most active samples.
Resazurin reduction assay (RRA) was used to determine the cytotoxicity of the studied samples. Cell cycle distribution (PI staining), apoptosis (annexin V/PI staining), mitochondrial membrane potential (MMP; JC-1) and reactive oxygen species (ROS; HDCFH-DA) were measured by flow cytometry. Column chromatography (CC) was used for the purification of PCF, whilst nuclear magnetic resonance (NMR) spectroscopic and mass spectrometric (MS) analyses were applied for structural elucidation.
The phytochemical investigation of PCF led to the isolation of 11 compounds: licarin B (1), licarin A (2), 7-(1,3-benzodioxol-5-yl)-7,8-dihydro-8-methyl-5-(2-propenyl)-furo[3,2-e]-1,3-benzodioxole (3), nitidine isocyanate (4), 5-hydroxy-7,4'-dimethoxyflavone (5), cardamomin (6), sitosterol (7) and stigmasterol (8), β-sitosterol 3-O-β--glucopyranoside (9), oleanolic acid (10) and lupeol (11). Fraction PCFb, compound 2 and doxorubicin (as positive control drug) revealed cytotoxic effects towards the 18 tested cancer cell lines. The IC values ranged from 6.1 μg/mL (against CCRF-CEM cells) to 44.2 μg/mL (against BRAF-V600E homozygous mutant melanoma cells) for PSCb; from 4.3 μM (against CCRF-CEM cells) to 21.8 μM (against HCT116 p53) for compound 2 and from 0.02 μM (against CCRF-CEM cells) to 123.0 μM (against CEM/ADR5000 cells) for doxorubicin. PCFb and compound 2 induced apoptosis in CCRF-CEM cells mediated by activation of caspase 3/7, 8 and 9, MMP alteration and increased ROS production.
Piper capense is a source of potent cytotoxic botanicals and phytochemicals that could help to fight various types of cancer including multidrug resistance phenotypes. PCFb and compound 2 should further be explored to develop new drugs to fight malignancies.
Piper capense 是一种药用香料，其果实传统上被用作水煎剂来治疗多种疾病，如锥虫病、寄生虫感染和癌症。
(1)对 Piper capense 的甲醇提取物进行植物化学研究；(2)评估植物药（PCF、PCFa-e 馏分）、分离出的植物化学物质对广泛的动物和人类癌细胞系的细胞毒性；(3)评估最活跃的样品诱导细胞凋亡的能力。
使用 Resazurin 还原测定法 (RRA) 来测定研究样品的细胞毒性。通过流式细胞术测量细胞周期分布 (PI 染色)、凋亡 (annexin V/PI 染色)、线粒体膜电位 (MMP；JC-1) 和活性氧 (ROS；HDCFH-DA)。柱层析 (CC) 用于 PCF 的纯化，而核磁共振 (NMR) 光谱和质谱 (MS) 分析则用于结构阐明。
PCF 的植物化学研究导致分离出 11 种化合物：利卡林 B（1）、利卡林 A（2）、7-(1,3-苯并二恶唑-5-基)-7,8-二氢-8-甲基-5-(2-丙烯基)-呋喃[3,2-e]-1,3-苯并二恶唑（3）、胡椒碱异氰酸酯（4）、5-羟基-7,4'-二甲氧基黄酮（5）、小豆蔻明（6）、豆甾醇（7）和谷甾醇（8）、β-谷甾醇 3-O-β--葡萄糖苷（9）、齐墩果酸（10）和羽扇豆醇（11）。PCFb 馏分、化合物 2 和阿霉素（阳性对照药物）对 18 种测试的癌细胞系表现出细胞毒性作用。IC 值范围从 6.1μg/mL（针对 CCRF-CEM 细胞）到 44.2μg/mL（针对 BRAF-V600E 纯合突变黑色素瘤细胞）对于 PSCb；从 4.3μM（针对 CCRF-CEM 细胞）到 21.8μM（针对 HCT116 p53）对于化合物 2，从 0.02μM（针对 CCRF-CEM 细胞）到 123.0μM（针对 CEM/ADR5000 细胞）对于阿霉素。PCFb 和化合物 2 通过激活 caspase 3/7、8 和 9、MMP 改变和增加 ROS 产生，诱导 CCRF-CEM 细胞凋亡。
Piper capense 是具有潜在细胞毒性的植物药和植物化学物质的来源，可帮助治疗各种类型的癌症，包括多药耐药表型。PCFb 和化合物 2 应进一步研究以开发用于治疗恶性肿瘤的新药。
