A method has been developed for the large scale isolation of 5-methoxypodophyllotoxin [1] from a high-producing root culture derived from Linum flavum. A closely related lignan, 5'-demethoxy-5-methoxypodophyllotoxin [2], was also present in the root culture and was the cause of the main isolation difficulties. Essential steps in the isolation procedure are CH2Cl2 and XAD-4 extraction and XAD-8 cc followed by Si gel chromatography, using two different mobile phases. The isolated 5-methoxypodophyllotoxin [1] was very pure (greater than 99%) and possessed the desired stereochemical configuration, namely (-)-5-methoxypodophyllotoxin [1]. The in vitro cytotoxicity of 5-methoxypodophyllotoxin [1] against EAT and HeLa cells was determined and compared with those of podophyllotoxin [3], etoposide (VP-16-213) [4], teniposide (VM-26) [5], and 5-methoxypodophyllotoxin-4-beta-D-glucoside [6]. It appeared that 5-methoxypodophyllotoxin [1] has about the same cytotoxic potency as podophyllotoxin [3].
Camptotheca acuminata plants contain camptothecin which is a secondary metabolite with strong anti-tumor activity. The induction of callus and cell suspension cultures from Camptotheca acuminata stem parts on different media is described. Growth in the original media and in media with different salts and/or vitamins was followed by measuring several growth parameters. Camptothecin was detected and identified by means of TLC, HPLC and GC-MS. The production of camptothecin in cell suspension cultures was followed and compared to the original plant material. Results indicate that high density cultures could be easily established, producing ca I mg 1-1 of camptothecin.
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