Cyclopamine

SKU: orb1304971

Description

Cyclopamine is a steroidal jerveratrum alkaloid that acts as a Smoothened (Smo) antagonist, with an IC50 of 46 nM in TM3Hh12 cells. It is a critical research tool for studying the Hedgehog signaling pathway in both in vitro and in vivo models of development and cancer.

Research Area

Cancer Biology, Cell Biology, Stem Cell & Developmental Biology

Key Properties

• CAS Number: 4449-51-8
• MW: 411.6
• Purity: >98%
• Formula: C₂₇H₄₁NO₂
• SMILES: [H][C@@]12CC3O[C@]4([H])C[C@H](C)CN[C@@]4([H])[C@@H](C)[C@@]3([H])C(C)=C1C[C@@]1([H])[C@@]2([H])CC=C2C[C@@H](O)CC[C@]12C
• Target: GPCR
• Solubility: Soluble in DMSO (up to 4 mg/ml) or in Ethanol (up to 20 mg/ml)

Bioactivity

• Target IC50:
  Smo:46 nM (TM3Hh12 cells)
• In Vivo:
  To examine the effects of cyclopamine treatment in vivo, subcutaneous xenografts from HUCCT1 cells, a metastatic cholangiocarcinoma cell line, were established in athymic mice. Tumours in cyclopamine-treated animals regressed completely by 12 days . In the delayed treatment model, no difference in weight was noted between control and cyclopamine (1.2 mg) treated BxPC3-SMOlow tumours. By contrast, a 50–60% decrease in tumour mass was observed in Panc 05.04- and L3.6sl-derived tumours, respectively (Fig. 5b, c)—an even more marked effect was noted in the concurrent treatment model, which revealed an 84% reduction in tumour mass of L3.6sl-derived tumours .
• In Vitro:
  The Chicken embryos Exposure to cyclopamine resulted in visible external defects, including cyclopia, microphthalmia, proboscis formation, amelia, thoracic lordosis, and decreased body size . cyclopamine treatment reduced the growth of tumor cell lines from the oesophagus, stomach, biliary tract and pancreas by 75–95% compared with tomatidine controls . In pancreatic cancer cell lines, Hh inhibition with cyclopamine resulted in down-regulation of snail and up-regulation of E-cadherin, consistent with inhibition of epithelial-to-mesenchymal transition, and was mirrored by a striking reduction of in vitro invasive capacity (P < 0.0001) .
• Cell Research:
  Cells were cultured in triplicate in 96-well plates in assay media to which 5E1 monoclonal antibody, ShhNp and/or cyclopamine were added at 0 h at concentrations indicated in the main text. Viable cell mass was determined by optical density measurements at 490 nm (OD490) at 2 and 4 days using the CellTiter96 colorimetric assay. Relative growth was calculated as OD (day 4) 2 OD (day 2)/OD (day 2) .
• Animal Research:
  A total of 0.1 ml Hanks' balanced salt solution and matrigel (1:1) containing 2 × 10^6 cells were injected subcutaneously into CD-1 nude mice. Tumours were grown for 4 days to a minimum volume of 125 mm3; treatment was initiated simultaneously for all subjects. Mice were injected subcutaneously with vector alone (triolein:ethanol 4:1 v/v) or a cyclopamine suspension (1.2 mg per mouse in triolein: ethanol 4:1 v/v) daily for 7 days. At the end of the treatment period, tumours were excised from mice, weighed and then fixed for 3 h at 4 °C with 4% paraformaldehyde, embedded in paraffin wax and sectioned (6 μm). Apoptotic cells were identified by TUNEL using recombinant Tdt as previously described29. Sections were then counterstained with eosin. Eight ×20-magnified fields from regions corresponding to the exterior, middle and interior of two control and two cyclopamine-treated tumours were chosen at random .

Storage & Handling

• Storage: -20°C
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

11-deoxojervine

Compound Identifiers

PubChem CID

52991976