五加皮中二萜类化学成分及其抗炎活性研究

Diterpenoids and their anti-inflammatory activities of Acanthopanacis Cortex

目的 对五加皮Acanthopanacis Cortex的二萜类化学成分及抗炎活性进行研究。方法 通过MCI树脂柱、硅胶柱、薄层色谱、半制备HPLC等方法对五加皮药材中化学成分进行提取分离,采用核磁共振波谱、质谱、红外光谱等方法,对单体化合物的结构进行鉴定。采用Griess法测试脂多糖诱导小鼠单核巨噬细胞RAW264.7细胞释放炎症因子模型,对化合物进行抗炎活性评价。结果 从五加皮70%乙醇提取物中分离得到13个二萜类化合物,包括7个对映海松烷型二萜7β-羟基-8(14),15-二烯-19-异海松烷酸(1)、7-酮基-8(14),15-二烯-19-异海松烷酸(2)、7β-甲氧基-8(14),15-二烯-19-异海松烷酸(3)、6,8(14),15-三烯-19-异海松羧酸(4)、14-酮基-8,15-二烯-19-异海松羧酸(7)、14-羟基-16-乙烯基-8,11,13-三烯-17-异海松羧酸(8)、7α-羟基-8(14),15-二烯-19-异海松烷酸(11);以及6个对映贝壳杉烷型二萜16α,17-二羟基-19-异贝壳杉烷羧酸(5)、17-羟基-19-异贝壳杉烷羧酸(6)、异贝壳杉烯酸(9)、17-醛基-19-异贝壳杉烯酸(10)、17-醛基-15-烯-19-异贝壳杉烷酸(12)和17-羟基-15-烯-19-异贝壳杉烷酸(13)。其中化合物8为新化合物,命名为细柱五加酸G1;3和4为首次从细柱五加植物中分离得到。体外抗炎活性研究表明,13个化合物均具有一定的抗炎活性,其中化合物1~3、7的活性较好。海松烷型二萜类化合物抗炎活性较贝壳杉烷型二萜强,海松烷型二萜类化合物7位取代活性强弱顺序为β-OH>β-OCH_(3)>Δ^(6)双键>C=O>α-OH。结论 五加皮分离鉴定的化合物及其活性研究为五加皮专属性质控方法建立、药用资源合理开发和利用提供了依据。

Objective To study diterpenoids and their anti-inflammatory activities of Wujiapi(Acanthopanacis Cortex,the root bark of Acanthopanax gracilistylus W.W.Smith).Methods The chemical constituents were separated and obtained by MCI resin column,silica gel column,thin-layer chromatography,semi-preparative HPLC and other methods.Their structures were identified by nuclear magnetic resonance,mass spectrometry,infrared spectrum,and so on.The Griess method was used to evaluate the anti-inflammatory activity by testing the inflammatory factor release model induced by lipopolysaccharide in mouse monocyte macrophage RAW264.7 cells.Results A total of 13 compounds were isolated from the 70%ethanol extract of Acanthopanacis Cortex,including seven entpimarane diterpenoids:7β-hydroxy-ent-pimara-8(14),15-dien-19-oic acid(1),7-oxo-ent-pimara-8(14),15-dien-19-oic acid(2),7β-methoxy-ent-pimara-8(14),15-dien-19-oic acid(3),ent-pimara-6,8(14),15-trien-19-oic acid(4),14-oxo-ent-pimara-8,15-dien-19-oicacid(7),14-hydroxy-16-devinyl-ent-pimara-8,11,13-trien-17-oic acid(8),7α-hydroxy-ent-pimara-8(14),15-dien-19-oic acid(11),and 6 ent-kaurene diterpenoids:16α,17-dihydroxy-ent-kaur-19-oic acid(5),17-hydroxy-ent-kaur-19-oic acid(6),ent-kaur-16-en-19-oic acid(9),17-oxo-ent-kaur-19-oic acid(10),17-oxo-ent-kaur-15-en-19-oic acid(12),and 17-hydroxy-ent-kaur-15-en-19-oic acid(13).Compound 8 is a new compound,named acangraciloic acid G1.Compound 3 and 4 are obtained from this plant for the first time.In vitro anti-inflammatory activity studies showed that all of the compounds exhibited anti-inflammatory activities,among which 1—3 and 7 showed higher anti-inflammatory activities.The anti-inflammatory activities of pimarane diterpenoids are stronger than those of kaurene diterpenoids.The activity order of C-7 substitution of pinane diterpenoids isβ-OH>β-OCH_(3)>Δ^(6) double bond>C=O>α-OH.Conclusion The compounds isolated and identified in this experiment and their activity research provide a basis for the establishment of specific quality control methods for Acanthopanacis Cortex,as well as the rational development and utilization of medicinal resources.