Sudha A, Srinivasan P
Department of Bioinformatics, Alagappa University, Karaikudi, Tamil Nadu, India.
Pharmacogn Mag. 2014 Aug;10(Suppl 3):S596-605. doi: 10.4103/0973-1296.139795.
Anisomeles malabarica R. Br. (Lamiaceae) is extensively used in traditional medicine in major parts of India for several medicinal purposes, including their use in rheumatism.
The air-dried leaves of A. malabarica were extracted with ethanol, defatted with n-hexane and then successively partitioned into chloroform and n-butanol fractions. Bioassay-guided fractionation and purification of chloroform fraction from A. malabarica lead to the isolation of lipoxygenase (LOX) inhibitors. The structures of isolated compounds were elucidated by ultraviolet, infrared, (1)H nuclear magnetic resonance (NMR), (13)C NMR and mass spectrometry spectroscopic techniques and assessed further by in vitro soybean lipoxygenase (sLOX) assay. In addition, the enzyme type inhibition was evaluated through molecular docking technique as a part of computational study.
The bioactive compounds 3, 4 dihydroxy benzoic acid (1) and 4', 5, 7-trihydroxyflavone (2) were isolated from chloroform fraction of A. malabarica, whose bioactivity was observed to be dose-dependent compared to n-butanol fraction. Among the compounds, 3, 4 dihydroxy benzoic acid showed significant sLOX inhibitory activity with 74.04% ±2.6% followed by 4', 5, 7-trihydroxyflavone (34.68% ±1.9%). The computational analysis of compounds showed their molecular interaction with important amino acid residues and nonheme iron atom in the catalytic site of LOX by enlightening their potential binding mode at molecular level.
The LOX inhibitory constituents were identified from A. malabarica by means of bioassay-guided fractionation process. The results derived from in vitro and computational experiments confirm the potential of the isolated compounds and provide additional evidence for its traditional use in inflammatory disorders.
印度薄荷（唇形科）在印度大部分地区被广泛用于传统医学，用于多种药用目的，包括治疗风湿病。
将印度薄荷的风干叶片用乙醇提取，用正己烷脱脂，然后依次分成氯仿和正丁醇馏分。通过生物测定指导的分级分离和纯化印度薄荷氯仿馏分，得到脂氧合酶（LOX）抑制剂。通过紫外、红外、（1）H核磁共振（NMR）、（13）C NMR和质谱光谱技术阐明分离化合物的结构，并通过体外大豆脂氧合酶（sLOX）测定进一步评估。此外，作为计算研究的一部分，通过分子对接技术评估酶类型抑制作用。
从印度薄荷氯仿馏分中分离出生物活性化合物3,4-二羟基苯甲酸（1）和4',5,7-三羟基黄酮（2），与正丁醇馏分相比，其生物活性呈剂量依赖性。在这些化合物中，3,4-二羟基苯甲酸显示出显著的sLOX抑制活性，为74.04%±2.6%，其次是4',5,7-三羟基黄酮（34.68%±1.9%）。化合物的计算分析通过揭示其在分子水平上的潜在结合模式，显示了它们与LOX催化位点中重要氨基酸残基和非血红素铁原子的分子相互作用。
通过生物测定指导的分级分离过程从印度薄荷中鉴定出LOX抑制成分。体外和计算实验的结果证实了分离化合物的潜力，并为其在炎症性疾病中的传统用途提供了额外证据。
