Department of Pharmacognosy and Phytochemistry, Faculty of Pharmacy, Universitas Airlangga, Jl. Mulyorejo, Surabaya, 60115, Indonesia.
Institute of Tropical Disease, Universitas Airlangga, Jl. Mulyorejo, Surabaya, 60115, Indonesia.
BMC Complement Med Ther. 2021 Jan 12;21(1):27. doi: 10.1186/s12906-021-03202-8.
New agents for developing alternative or complementary medicine to treat the hepatitis C virus (HCV) are still needed due to high rates of HCV infection globally and the current limitations of available treatments. Treatment of HCV with a combination of direct acting antivirals have been shown to be approximately 90% effective but will be limited in the future due to the emergence of drug resistance and high cost. The leaves of Melicope latifolia have previously been reported to have anti-HCV activity and are a potential source of bioactive compounds for future novel drug development. This study aimed to evaluate the efficacy of the extract of M. latifolia fruit to treat HCV and to isolate its active compounds.
M. latifolia fruit was extracted using methanol and purified using vacuum liquid chromatography (VLC) and Radial Chromatography. The anti-HCV activity was analyzed using cell culture lines Huh7it-1 and JFH1 (genotype 2a). Time-of-addition and immunoblotting studies were performed to identify the mode of action of the isolated active compounds. The structures of the active compounds were determined using nuclear magnetic resonance (NMR) spectra, UV, IR, and Mass Spectra.
Six known compounds were isolated from M. latifolia fruit: O-methyloktadrenolon, alloevodionol, isopimpinellin, alloxanthoxyletin, methylevodionol, and N-methylflindersine. N-methylflidersine was the most active compound with IC value of 3.8 μg/ml while methylevodionol, isopimpinellin, and alloevodionol were less active. O-methyloktadrenolon and alloxanthoxyletin were moderately active with IC values of 10.9 and 21.72 μg/ml, respectively. N-methylflidersine decreased level of HCV NS3 protein expression in the cells.
The alkaloid compound, N-methylflindersine which was isolated from M. latifolia possesses anti-HCV activity through post-entry inhibition and suppressed NS3 protein expression.
由于全球 HCV 感染率高，以及现有治疗方法的局限性，仍需要开发替代或补充医学的新药物来治疗丙型肝炎病毒 (HCV)。使用直接作用抗病毒药物联合治疗 HCV 的效果约为 90%，但由于耐药性的出现和成本高昂，未来这种治疗方法将受到限制。此前有报道称，桃金娘科植物桃金娘的叶子具有抗 HCV 活性，是未来新型药物开发的潜在生物活性化合物来源。本研究旨在评估桃金娘果实提取物治疗 HCV 的疗效，并分离其活性化合物。
采用甲醇提取桃金娘果实，采用真空液相色谱（VLC）和径向色谱法进行纯化。采用细胞培养系 Huh7it-1 和 JFH1（基因型 2a）分析抗 HCV 活性。进行时间添加和免疫印迹研究，以确定分离出的活性化合物的作用方式。使用核磁共振（NMR）谱、UV、IR 和质谱确定活性化合物的结构。
从桃金娘果实中分离得到 6 种已知化合物：O-甲基 oktadrenolon、alloevodionol、异补骨脂素、alloxanthoxyletin、甲氧基 evodionol 和 N-甲基 flindersine。N-甲基 flindersine 是最有效的化合物，IC 值为 3.8μg/ml，而甲氧基 evodionol、异补骨脂素和 alloevodionol 的活性较低。O-甲基 oktadrenolon 和 alloxanthoxyletin 的 IC 值分别为 10.9 和 21.72μg/ml，活性中等。N-甲基 flindersine 降低了细胞内 HCV NS3 蛋白的表达水平。
从桃金娘中分离得到的生物碱化合物 N-甲基 flindersine 通过进入后抑制作用发挥抗 HCV 活性，并抑制 NS3 蛋白的表达。
