Chemical Constituents Of Machilus Ichangensis Rehd. Et Wils. And Their Biological Activities

Machilus ichangensis Rehd. et Wils.(Lauraceae) belongs to macrophanerophytes, which is widely distributed in the broad-leafed forests in Hubei, Sichuan, southern Shaanxi and Western Gansu Provinces. This plant was recorded in Chinese traditional and folk medicine books, mainly for treatments of various skin inflammations, swelling of the joints and rheumatic diseases in the Three Gorges Region, having significant effects, but no obvious toxic side effects. However, no chemical constituents or biological activities relating to this plant have been reported.In this paper, chemical constituents of Machilus ichangensis, collected from Dalaoling in Hubei Province, was investigated, in order to provide a scientific basis for the further research and development of this genus. According to its folk application characteristics and pharmacology literature, we evaluated the pharmacological activities of its extracts and compounds, using in vitro cytotoxicity, neuroprotective and anti-inflammatory models.The main results can be described as follows:1. Various chromatographic methods were used for the isolation of the compounds, and their structures were identified by modern spectral technologies (including IR, UV, MS,’H NMR,13C NMR, DEPT,2D NMR and CD) with physicochemical properties. Fifty-two compounds were isolated from the95%ethanol extract of the stems of Machilus ichangensis, and the structures of them were identified as follows:machilusmarin (1), machilusolide A (2), machilusolide B (3), machilusolide C (4), machilusolide D (5), machilusolide E (6), machilusolide F (7),(-)-licarin A (8),(-)-acuminatin (9), odoratisol A (10),(-)-licarin B (11),(+)-pinoresinol (12),(+)-syringaresinol (13),(+)-medioresinol (14),4-ketopinoresinol (15),iso-4-ketopinoresinol (16),(-)-lyoniresinol-3α-O-β-D-glucopyranoside (17), (-)-5’-methoxyisolariciresinol-3α-O-β-D-glucopyran-oside(18), isolariciresinol-9-O-β-D-glucopyranoside (19), nectandrin B (20), nectandrin A (21), galgravin (22), fragransins B3(23),(-)-secoisolariciresinol (24), machilin A (25), scopoletin (26), isoscopoletin (27), E-p-coumaric acid (28), caffeic acid (29), ferulic acid (30), isoerulic acid (31), elemicin (32), chavico-l-O-β-D-glucopyranoside (33),(+)-catechin (34),(-)-epicatechin (35),(2R,3R)-4’-hydroxy-5,7,3’-trimethoxyflavan-3-ol (36),(2R,3R)-3-hydroxy-5,7-dimethoxy-3’,4’-methylenedioxyflavan (37), naringenin (38), davidigenin (39), piperonylic acid (40), vanillic acid (41), protocatechuic acid (42), syringic acid (43),3,4,5-trimethoxyphenol-l-O-β-D-glucopyranoside (44), phenolphthalein (45), icariside B2(46), byzantionosides A (47),(E)-4-(1,2,4-trihydroxy-2,6,6-trimethylcyclohexyl)-but-3-en-2-one (48), abscisic acid (49),β-sitosterol (50), daucosterol (51), and palmitic acid (52), including six butanolides, eighteen lignans, three coumarins, six phenylpropanoids, six flavonoids, six phenols, four β-ionones, two steroids and one fatty acids. Among them, seven compounds were new, which included one dimeric isocoumarin derivate (1) and six butanolides (2-7). All those compounds were isolated from this plant for the first time.2. In vitro cytotoxicity of compounds was determined by MTT assay. Six new butanolides showed selective cytotoxicity against MDA-MB-231, compound3had the best cytotoxicity, with IC503.22μM; The neuroprotective activity of all the compounds was evaluated by SH-SY5Y human neuroblastoma cells apoptosis induced by hydrogen peroxide. Compound1showed significant neuroprotective activity against SH-SY5Y human neuroblastoma cells apoptosis induced by hydrogen peroxide, its concentrations ranging from0.625μg/mL to10.0μg/mL; Inflammation model in vitro was established by stimulating human acute monocytic leukemia cell line with lipopolysaccharide, after which were differentiated into macrophages by phorbol ester. Cytotoxicity of total glucoside of Machilus ichangensis was determined by MTT assay, the anti-inflammatory potential of total glucoside of Machilus ichangensis was evaluated by monitoring its effects on the modulation of cytokines and mediators of inflammation, such as interleukin1β, interleukin6, tumor necrosis factor-a and nitric oxide in cell culture medium, using ELISA methods. Total glucoside of Machilus ichangensis showed great significant dose-and time-dependent inhibitory activities on IL-6, TNF-a and NO synthesis, but slight effect on IL-1β.