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


CONCLUSIONS
This method could meet a very real need within diverse scientific communities by allowing cyclopamine to become more readily available.
Oatis
John E
JE
Department of Pharmacology & Experimental Therapeutics, Medical University of South Carolina, Charleston, SC 29425 USA. oatisj@musc.edu
Brunsfeld
Pam
P
Rushing
James W
JW
Moeller
Peter D
PD
Bearden
Daniel W
DW
Gallien
Thomas N
TN
Cooper
George
G