Isolation, Purification and Characterization of Novel Steroidal Alkaloids from \u3cem\u3eVeratrum californicum\u3c/em\u3e

Cyclopamine is a steroidal alkaloids in Veratrum californicum that is a teratogen. Steroidal alkaloids from this plant have been shown to inhibit the Sonic hedgehog (Shh) signaling pathway, resulting in embryo deformities including cyclopia in lambs. The Shh signaling pathway is prevalent in over 20 types of cancer, and contributes to the overproduction of cancerous cells and tumor growth. Qualitative and quantitative analysis of Veratrum californicum alkaloids has confirmed various abundancies of cyclopamine and other alkaloids in different sections of the plant (leaf, stem, root/rhizome), with the highest amount of alkaloid present in the root and rhizome. Shh Light II cells provide a colorimetric assay to assess the degree of Shh pathway inhibition by chemical agents. This assay was used to evaluate alkaloid ratios by plant part. The results showed the greatest pathway inhibition was achieved by the ratio of steroidal alkaloids consistent with that derived from the root and rhizome portion of the plant, followed by stem, and finally leaf. Further analysis of the root and rhizome extract by high pressure liquid chromatography and mass spectrometry has verified the presence of uncharacterized, novel alkaloids that may be potent Shh signaling pathway antagonists. The focus of the current work is to extract, isolate and characterize novel alkaloids and evaluate their bioactivity using the Shh Light II cell assay