Isolation, purification, and full NMR assignments of cyclopamine from Veratrum californicum

• Published in: Chemistry Central journal Vol. 2; no. 1; p. 12
• Main Authors: Oatis, John E, Brunsfeld, Pam, Rushing, James W, Moeller, Peter D, Bearden, Daniel W, Gallien, Thomas N, Cooper, George
• Format: Journal Article
• Language: English
• Published: Cham Springer International Publishing 24.06.2008; BioMed Central Ltd; BioMed Central; BMC
• Subjects: Analytical Chemistry; Chemistry; Chemistry and Materials Science; Chemistry/Food Science; Methodology; Steroidal Alkaloid; Partial Assignment; Flash Chromatography; Aqueous Formic Acid; Cyclopamine
• ISSN: 1752-153X; 1752-153X
• DOI: 10.1186/1752-153X-2-12

Background The Hedgehog signaling pathway is essential for embryogenesis and for tissue homeostasis in the adult. However, it may induce malignancies in a number of tissues when constitutively activated, and it may also have a role in other forms of normal and maladaptive growth. Cyclopamine, a naturally occurring steroidal alkaloid, specifically inhibits the Hedgehog pathway by binding directly to Smoothened, an important Hedgehog response element. To use cyclopamine as a tool to explore and/or inhibit the Hedgehog pathway in vivo , a substantial quantity is required, and as a practical matter cyclopamine has been effectively unavailable for usage in animals larger than mice. Results In this paper, we report a rapid and efficient isolation and purification of large quantities of cyclopamine from the roots and rhizomes of Veratrum californicum Dur. (the Corn Lily or Western false hellebore). We also provide unambiguous assignments of the carbon and proton resonances by using the multinuclear spectra and the spin coupling networks. Conclusion This method could meet a very real need within diverse scientific communities by allowing cyclopamine to become more readily available.