蟾皮中的蟾蜍甾烯类成分及其细胞毒活性研究

Cytotoxic bufadienolides of skin of Bufo bufo gargarizans

目的研究蟾皮(中华大蟾蜍Bufo bufo gargarizans的干燥皮)中的蟾蜍甾烯类成分及其细胞毒活性。方法利用硅胶、ODS、凝胶、高效液相色谱等方法对中药蟾皮的化学成分进行分离,运用IR、UV、NMR、MS及单晶X-衍射等波谱学方法对分离得到的化合物进行结构鉴定,采用MTT法测试化合物对人肝癌HepG2细胞、人非小细胞肺癌A549细胞及人宫颈癌He La细胞的增殖抑制活性。结果从蟾皮95%乙醇提取物的氯仿萃取部位共分离鉴定了14个蟾蜍甾烯类成分,分别为3,23-二羟基-γ-内酯-24-去甲-14,20(22)-二烯-21-胆酸(1)、3-表-酯蟾毒配基(2)、3-表-去乙酰华蟾蜍精(3)、3-表-蟾蜍灵(4)、3-氧-去乙酰华蟾蜍精(5)、3-氧-蟾蜍它灵(6)、3-氧-伪沙蟾蜍精(7)、3-oxo-Δ4(5)-resibufogenin(8)、3-oxo-Δ4(5)-cinobufotalin(9)、3-dehydroscillarenin(10)、3-氧-毛地黄毒苷配基(11)、digirezigenin(12)、bufogargarizin A(13)、bufogargarizin B(14)。化合物6、9、10对Hep G2、A549及He La细胞具有显著的增殖抑制活性,半数抑制浓度(IC50)为0.11~1.67μmol/L。结论化合物1为新天然产物,并首次报道其核磁数据;化合物1~14均为首次从蟾皮中分离得到;化合物6、9、10具有显著的细胞毒活性。

Objective To investigate the cytotoxic bufadienolides from the skin of Bufo bufo gargarizans.Methods Compounds were isolated by a combination of various chromatographic methods including silica gel,ODS and HPLC,and their structures were elucidated based on spectroscopic analyses(IR,UV,NMR,and MS)and single crystal X-ray diffraction.The in vitro cytotoxic activities of 10 bufadienolides were evaluated in HepG2,A549,and Hela cells by MTT assay.Results A total of 14 compounds were isolated from the skin of B.bufo gargarizans and identified as 3,23-dihydroxy-γ-lactone-24-norchola-14,20(22)-dien-21-oicacid(1),3-epi-resibufogenin(2),3-epi-desacetylcinobufagin(3),3-epi-bufalin(4),3-ketodesacetylcinobufagin(5),3-oxocinobufagin(6),3-oxo-ψ-bufarenogin(7),3-oxo-Δ4(5)-resibufogenin(8),3-oxo-Δ4(5)-cinobufotalin(9),3-dehydroscillarenin(10),3-dehydrodigitoxigenin(11),digirezigenin(12),bufogargarizin A(13),and bufogargarizin B(14).Three compounds(6,9,and 10)displayed significant anti-proliferative activities on HepG2,A549 and Hela cells with IC50 values of 0.11-1.67μmol/L.Conclusion Compound 1 is a new natural product.Compounds 1-14 are reported for the first time in the skin of B.bufo gargarizans.Compounds 6,9,and 10 show significant cytotoxic activities.