Study On In Vitro Antitumor Constituents From Two Medicinal Plants

Cancer is an important threat to human health,so it attached great significance to find the drugs that exhibited the anti-tumor activities.Modern pharmacological studies displayed that many traditional Chinese medicines have strong antitumor activities,and have been widely used in clinic.Therefore,clarifying the material bases and finding lead compounds revealed antitumor activities from medicinal materials,are a hot topic in traditional Chinese medicine research.In this paper,we selected two kinds of medicinal materials,Mirabilis himalaica and Corydalis tomentella Franch.And our ongoing efforts on the chemical investigation of the extraction site with antitumor activities were studied systematically.1.Study on in vitro antitumor constituents from Mirabilis himalaica.The roots of Mirabilis himalaica is one of the most frequently used folk medicine in Tibet,China for the treatment of uterine cancer,nephritis edematous,renal calculus and arthrodynia.It was reported that Mirabilis himalaica exhibited anti-tumor activities in vitro.Moreover,previously chemical investigations on the roots of Mirabilis himalaica indicated that 95%ethanol extract displayed inhibitory effects on A549 and HCT-8 cells.And ethyl acetate extract exhibited anti-tumor effects significantly in vitro.In order to further clarify the material basis of antitumor effect,the chemical composition of ethyl acetate extract of Mirabilis himalaica was studied systematically.Moreover,the cytotoxic activities of all isolated compounds were assayed against human lung carcinoma A549 cells and human hepatocellular carcinoma Hep G2 cells in vitro.Kinds of chromatographic methods including normal phase silica gel column chromatography,Sephadex LH-20,semi-preparative HPLC,recrystallization were applied for the isolation and purification of the ethyl acetate soluble extract.Their structures were elucidated by spectroscopic experiments including ¹H-NMR、¹³C-NMR、2D-NMR、CD、UV、IR,and combined HR-ESI-MS and reported papers.Finally,eighteen known ones were obtained,and identified as himalaflavone A（L-1）,himalaflavone B（L-2）,himalaflavone C（L-3）,himalaflavone D（L-4）,himalaflavone E（L-5）,3,7,4′-trihydroxy-5-methoxy-6-methylflavanone（L-6）,（2S）-7-hydroxy-5-methoxy-6-methylflavanone（L-7）,pisonivanone（L-8）,（2S）-7-hydroxy-5-methoxy-6,8-dimethylflavanone（L-9）,6-methyl-（2R,3S）-alpinone（L-10）,2′,3,4′,5,7-pentahydroxy-6-methylflavone（L-11）,abronione（L-12）,mirabijalone A（L-13）,10-demethyl boeravinone C（L-14）,and boeravinone A methyl ether（L-15）,caffeic acid（L-16）,dodecylic acid（L-17）,hexadecyl haffeate（L-18）,including 15 flavonoids,2 phenylpropanoids and 1 organic acid.Moreover,L-1～5 were five previously undescribed flavonoids and L-6～11,14,15,17 and 18 were isolated from this plant for the first time.The cytotoxic activities of all isolated compounds were assayed against human tumor cells in vitro,and compounds L-4,5,11 and 13 exhibited inhibitory effects on A549 and compounds L-8 and 11 exhibited inhibitory effects on Hep G2 cells.2.Study on chemical constituents from Corydalis tomentella Franch.Corydalis tomentella Franch（Papaveraceae）is costly Chinese folk medicine in the southwest,China with the effects of clearing heat,detoxifying,cooling blood and liminating stasis to subdue swelling for treatment of swelling and pain in throat andocular region,liver-depressed hypochondrium pain,madidans and diarrhea and carbuncle swelling.Corydalis tomentella Franch has shown the strongly activities,such as anti-inflammatory,antibiosis,analgesia,and antitumor etc.Moreover,it is used as a hepatoprotective herb to treat hepatitis,liver cirrhosis and liver cancer.In our study,kinds of polar extracts were assayed against Hep G2 cells in vitro.The ethyl acetate soluble extract exhibited the most significantly inhibitory effects was futher studied systematically,in order to obain chemical constituents of possessing novel structures and high activities.Isolation and purification of the ethyl acetate soluble extract using normal phase silica gel column chromatography,reversed phase silica gel column chromatography,Sephadex LH-20,MCI,semi-preparative HPLC,medium-pressure preparative chromatography,recrystallization,etc.Their structures were elucidated by spectroscopic experiments including ¹H-NMR、¹³C-NMR and reported papers.Finally,seventeen known compounds obtained from Corydalis tomentella Franch were determined as rupestrine A（Et-1）,cheilanthifoline（Et-2）,dehydrocorydaline（Et-3）,isocorybulbine（Et-4）,bicuculline（Et-5）,corybulbine（Et-6）,palmatine（Et-7）,trans-N-feruloyloctopamine（Et-8）,corynoxidine（Et-9）,（-）-trans-Isocorypalmine N-oxide（Et-10）,epicorynoxidine（Et-11）,（-）–corynoxidine（Et-12）,corydaldine（Et-13）,oxyhydrastinine（Et-14）,noroxyhydrastinine（Et-15）,doryanine（Et-16）,7-epiloliolide（Et-17）,including 16 alkaloids and 1 terpenoid.Among them,Et-1～4,6～13,15～17 were isolated from this plant for the first time.The chemical investigation of the ethyl acetate extract from Mirabilis himalaica and Corydalis tomentella Franch were studied systematically.18 and 17 ones were isolated from the two plants,respectively.Among them,including five previously undescribed flavonoids.The cytotoxic activities of all isolated compounds from Mirabilis himalaica were assayed against tumor cells.The results showed that six compounds exhibited inhibitory effect against A549 cells or Hep G2 cells.The present study has important theoretical value and practical significance for enriching the chemical composition library of the plants,finding the leading compounds against tumor,and laying a material foundation for the comprehensive development and utilization of medicinal materials.