Cyclopamine

製品コードS1146 バッチS114612

印刷

化学情報

Chemical Structure Synonyms 11-deoxojervine Storage
(From the date of receipt)		 3 years -20°C powder
1 years -80°C in solvent
化学式

C27H41NO2
分子量 411.62 CAS No. 4449-51-8
Solubility (25°C)* 体外 Ethanol (warmed with 50ºC water bath) 21 mg/mL (51.01 mM)
DMSO Insoluble
Water Insoluble
体内 （毎回新しく調製した物を用意してください）
Clear solution
5%Ethanol 40%PEG300 5%Tween80 50%ddH2O

この製剤はselleckのラボで検証済みです。上記の溶解方法がご要望を満たさない場合、selleckの営業担当までお問い合わせ頂ければ、個別の試験を行います。

 1.000mg/ml (2.43mM) Taking the 1 mL working solution as an example, add 50 μL 20 mg/ml clarified Ethanol stock solution to 400 μL PEG300, mix evenly to clarify it; add 50 μL Tween80 to the above system, mix evenly to clarify it; then continue to add 500 μL ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results. 
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

溶剤液（一定の濃度）を調合する

生物活性

製品説明 Cyclopamine (11-deoxojervine) is a specific Hedgehog (Hh) signaling pathway antagonist of Smoothened (Smo) with IC50 of 46 nM in TM3Hh12 cells.
in vitro Cyclopamine inhibits the Hedgehog signaling pathway with an IC50 of 46 nM, and blocks the activity of human Smo receptor expressed in CHO-K1 cells in [3H]Hh-Ag binding assay with an IC50 of 280 nM. [1] This compound significantly inhibits Hedgehog pathway activity in a dose-dependent manner in gut-derived tumor cell lines expressing Patched (PTCH) mRNA, and induces growth inhibition of those tumor cell lines by 75-95% at the concentration of 3 μM, but ineffective towards the colon tumor cells without PTCH mRNA expression, suggesting the effects of this chemical treatment are Hedgehog pathway related rather than generally cytotoxic. [2] By blocking Hedgehog signaling through direct interaction with Smo, it (10 μM) inhibits the proliferation of SMOhigh Cyclopamine-responsive cell lines L3.6sl and Panc 05.04 by 75-80%, and increases the apoptosis by 2.5- to 3.5-fold, without affecting the BxPC3-SMOlow cell line. [3] This treatment significantly decreases of Snail mRNA and increasea E-cadherin transcripts in the E3LZ10.7 cell line. Independent of inhibition of cell growth, it significantly inhibits the invasive phenotype of Hedgehog-dependent L3.6pl cells, causing a >500-fold reduction in the number of transmigrating cells, but not that of the Hedgehog-independent cell line Panc-1. [4]
in vivo Administration of Cyclopamine at dose of 50 mg/kg/day for 22 days eradicates the HUCCT1 xenografts in mice with no obvious adverse effects. [2] This compound treatment at dose of 1.2 mg for 7 days induces significant apoptosis of tumor cells and decreases the tumor mass by 50-60% in Panc 05.04- and L3.6sl-derived tumors, respectively, but not in the BxPC3-SMOlow tumors. [3] Administration of this chemical alone profoundly inhibits tumor metastases in xenografts of E3LZ10.7 and L3.6pl, and completely abrogates metastases when in combination of gemcitabine. [4]

プロトコル(参考用のみ)

キナーゼアッセイ Hedgehog cell assay
This assay measures the end stage of the Hh signaling pathway, that is, the transcriptional modulation of Gli, using Luciferase as readout (Gli-Luc assay). Cyclopamine is prepared for assay by serial dilution in DMSO and then added to empty assay plates. TM3Hh12 cells (TM3 cells containing Hh-responsive reporter gene construct pTA-8xGli-Luc) are resuspended in F12 Ham's/DMEM (1:1) containing 5% FBS and 15 mM Hepes pH 7.3, added to assay plates and incubated with this compound for approximately 30 minutes at 37 °C in 5% CO2. 1 nM Hh-Ag 1.5 is then added to assay plates and incubated at 37 °C in the presence of 5% CO2. After 48 hours, either Bright-Glo or MTS reagent is added to the assay plates and luminescence or absorbance at 492 nm is determined. IC50 value, defined as the inflection point of the logistic curve, is determined by non-linear regression of the Gli-driven luciferase luminescence or absorbance signal from MTS assay vs log10 (concentration) of this chemical using the R statistical software pack
細胞アッセイ 細胞株 SEG1, OE33, KYAE, KYSE180, SNU1, AGS, SNU16, NCI-N-87, HUCCT1, PANC1, PL5, PL6, BXPC3, HS766T, KYSE150, GBD1, DLD1, and HCT116
濃度 Dissolved in DMSO, final concentration 3 μM
反応時間 4 days
実験の流れ Cells are exposed to Cyclopamine in 96-well plates. Cell viability is measured by MTS (soluble tetrazolium salt) assay. Viable cell mass is determined by optical density measurements at 490 nm (OD490) at 2 and 4 days using the CellTiter96 colorimetric assay. Relative growth is calculated as OD (day 4)﹣OD (day 2)/OD (day 2).
動物実験 動物モデル Athymic (nude) mice inoculated subcutaneously with HUCCT1 cells
投薬量 50 mg/kg/day
投与方法 Subcutaneous injection

参考

  • https://pubmed.ncbi.nlm.nih.gov/19091559/
  • https://pubmed.ncbi.nlm.nih.gov/14520411/
  • https://pubmed.ncbi.nlm.nih.gov/14520413/
  • https://pubmed.ncbi.nlm.nih.gov/17332349/

カスタマーフィードバック

Data from [Cancer Res, 2012, 72, 2262-74]

Data from [Cancer Lett, 2012, 322, 169-176]

Data from [Exp Hematol, 2012, 40, 418-27]

Selleckの高級品が、幾つかの出版された研究調査結果（以下を含む）で使われた：

The O-glycosyltransferase C1GALT1 promotes EWSR1::FLI1 expression and is a therapeutic target for Ewing sarcoma [ Nat Commun, 2025, 16(1):1267] PubMed: 39894896
PlexinD1 is a driver and a therapeutic target in advanced prostate cancer [ EMBO Mol Med, 2025, 17(2):336-364] PubMed: 39748059
PPARG contributes to urothelial integrity in the murine ureter by activating the expression of Shh and superficial cell-specific genes [ Development, 2025, 152(8)dev204324] PubMed: 40167323
Interplay of SHH, WNT and BMP4 signaling regulates the development of the lamina propria in the murine ureter [ Development, 2025, 152(3)DEV204214] PubMed: 39817691
SHH Pathway Inhibition and Astrocyte Co-culture Induce Distinct Responses in Glioblastoma and Cancer Stem Cells [ Res Sq, 2025, rs.3.rs-7214243] PubMed: 40894044
Proximity based proteomics reveals Git1 as a regulator of Smoothened signaling [ bioRxiv, 2025, 2025.01.06.631593] PubMed: 39829937
Foxo3-mediated physiological cell competition ensures robust tissue patterning throughout vertebrate development [ Nat Commun, 2024, 15(1):10662] PubMed: 39690179
Phosphorylation of human glioma-associated oncogene 1 on Ser937 regulates Sonic Hedgehog signaling in medulloblastoma [ Nat Commun, 2024, 15(1):987] PubMed: 38307877
Establishment, characterization, and biobanking of 36 pancreatic cancer organoids: prediction of metastasis in resectable pancreatic cancer [ Cell Oncol (Dordr), 2024, 10.1007/s13402-024-00939-5] PubMed: 38619751
Transcription of microRNAs is regulated by developmental signaling pathways and transcription factors [ Front Cell Dev Biol, 2024, 12:1356589] PubMed: 38721525

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人間や獣医の診断であるか治療的な使用のためにでない。

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