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

John E Oatis Jr¹ , Pam Brunsfeld² , James W Rushing³ , Peter D Moeller⁴ , Daniel W Bearden⁵ , Thomas N Gallien⁶ and George Cooper IV⁶

¹Department of Pharmacology & Experimental Therapeutics, Medical University of South Carolina, Charleston, SC 29425 USA

²University of Idaho Stillinger Herbarium, Moscow, ID 83844 USA

³Clemson University Coastal Research & Education Center, Charleston, SC 29414 USA

⁴Toxin/Natural Products Program, Center for Coastal Environmental Health & Biomolecular Research, National Oceanic & Atmospheric Administration – National Ocean Service, Charleston, SC 29412 USA

⁵National Institute of Standards and Technology, Hollings Marine Laboratory, Charleston, SC 29412 USA

⁶Gazes Cardiac Research Institute, Cardiology Division, Medical University of South Carolina, Charleston, SC 29403 USA

author email corresponding author email

Chemistry Central Journal 2008, 2:12doi:10.1186/1752-153X-2-12


Published:	24 June 2008

Abstract

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.