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Cyclopamine (11-Deoxojervine) Hedgehog Signaling Pathway Antagonist

Name: Cyclopamine (11-Deoxojervine)
Brand: Selleck Chemicals
SKU: S1146
Rating: 5 (146 reviews)

Cat.Nr.S1146

Cyclopamine (11-deoxojervine) is een specifieke Hedgehog (Hh) signaleringsrouteantagonist van Smoothened (Smo) met een IC50 van 46 nM in TM3Hh12-cellen.

Cyclopamine (11-Deoxojervine) Hedgehog/Smoothened antagonist Chemical Structure

Chemische structuur

Moleculair gewicht: 411.62

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Kwaliteitscontrole

Batch: Zuiverheid: 99.99%

99.99

Gerelateerde doelwitten	JAK TGF-beta/Smad Wnt/beta-catenin ERK GSK-3 ROCK PKA Secretase STAT Casein Kinase
Overige Hedgehog/Smoothened Inhibitoren	SAG (Smoothened Agonist) Hydrochloride Purmorphamine GANT61 SAG (Smoothened Agonist) SANT-1 HPI-4 (Ciliobrevin A) BMS-833923 Taladegib (LY2940680) Ciliobrevin D PF-5274857

Celkweek, behandeling & werkconcentratie

Cellijnen	Assaytype	Concentratie	Incubatietijd	Activiteitsbeschrijving	PMID
OS-RC-2	Growth Inhibition Assay			IC50=5.8666 μM	SANGER
DOHH-2	Growth Inhibition Assay			IC50=9.35689 μM	SANGER
no-10	Growth Inhibition Assay			IC50=9.9039 μM	SANGER
LS-513	Growth Inhibition Assay			IC50=11.3547 μM	SANGER
ALL-PO	Growth Inhibition Assay			IC50=11.7734 μM	SANGER
8-MG-BA	Growth Inhibition Assay			IC50=13.1123 μM	SANGER
RPMI-8402	Growth Inhibition Assay			IC50=15.8537 μM	SANGER
EoL-1-cell	Growth Inhibition Assay			IC50=18.5948 μM	SANGER
NALM-6	Growth Inhibition Assay			IC50=19.0167 μM	SANGER
DEL	Growth Inhibition Assay			IC50=20.1471 μM	SANGER
SR	Growth Inhibition Assay			IC50=23.6715 μM	SANGER
697	Growth Inhibition Assay			IC50=26.6155 μM	SANGER
COLO-829	Growth Inhibition Assay			IC50=26.8483 μM	SANGER
EVSA-T	Growth Inhibition Assay			IC50=27.5561 μM	SANGER
ATN-1	Growth Inhibition Assay			IC50=31.2329 μM	SANGER
L-363	Growth Inhibition Assay			IC50=31.7461 μM	SANGER
LAMA-84	Growth Inhibition Assay			IC50=32.5211 μM	SANGER
NOS-1	Growth Inhibition Assay			IC50=34.2956 μM	SANGER
BB30-HNC	Growth Inhibition Assay			IC50=34.3306 μM	SANGER
BC-1	Growth Inhibition Assay			IC50=37.9746 μM	SANGER
IST-SL2	Growth Inhibition Assay			IC50=38.224 μM	SANGER
D-392MG	Growth Inhibition Assay			IC50=40.2215 μM	SANGER
no-11	Growth Inhibition Assay			IC50=40.5521 μM	SANGER
LC4-1	Growth Inhibition Assay			IC50=40.8716 μM	SANGER
A388	Growth Inhibition Assay			IC50=42.5848 μM	SANGER
NTERA-S-cl-D1	Growth Inhibition Assay			IC50=42.7074 μM	SANGER
CESS	Growth Inhibition Assay			IC50=44.2232 μM	SANGER
RS4-11	Growth Inhibition Assay			IC50=49.0938 μM	SANGER
MS-1	Growth Inhibition Assay			IC50=50.9351 μM	SANGER
CTV-1	Growth Inhibition Assay			IC50=51.074 μM	SANGER
D-502MG	Growth Inhibition Assay			IC50=51.6271 μM	SANGER
ML-2	Growth Inhibition Assay			IC50=52.9195 μM	SANGER
SK-NEP-1	Growth Inhibition Assay			IC50=53.3923 μM	SANGER
LOXIMVI	Growth Inhibition Assay			IC50=53.5884 μM	SANGER
DJM-1	Growth Inhibition Assay			IC50=56.3391 μM	SANGER
GI-1	Growth Inhibition Assay			IC50=56.6149 μM	SANGER
IST-MES1	Growth Inhibition Assay			IC50=60.5493 μM	SANGER
MV-4-11	Growth Inhibition Assay			IC50=60.6538 μM	SANGER
OVCAR-4	Growth Inhibition Assay			IC50=63.5657 μM	SANGER
KE-37	Growth Inhibition Assay			IC50=66.2668 μM	SANGER
D-542MG	Growth Inhibition Assay			IC50=68.4135 μM	SANGER
MHH-PREB-1	Growth Inhibition Assay			IC50=72.8441 μM	SANGER
MRK-nu-1	Growth Inhibition Assay			IC50=73.4705 μM	SANGER
D-247MG	Growth Inhibition Assay			IC50=73.5442 μM	SANGER
OCI-AML2	Growth Inhibition Assay			IC50=76.9369 μM	SANGER
LP-1	Growth Inhibition Assay			IC50=82.8731 μM	SANGER
HCC1599	Growth Inhibition Assay			IC50=84.2837 μM	SANGER
KARPAS-45	Growth Inhibition Assay			IC50=84.6992 μM	SANGER
BE-13	Growth Inhibition Assay			IC50=99.0477 μM	SANGER
GCIY	Growth Inhibition Assay			IC50=99.0954 μM	SANGER
BV-173	Growth Inhibition Assay			IC50=100.325 μM	SANGER
LB2518-MEL	Growth Inhibition Assay			IC50=100.789 μM	SANGER
KS-1	Growth Inhibition Assay			IC50=101.639 μM	SANGER
MOLT-16	Growth Inhibition Assay			IC50=104.986 μM	SANGER
NCI-H1770	Growth Inhibition Assay			IC50=108.784 μM	SANGER
NCI-H82	Growth Inhibition Assay			IC50=110.976 μM	SANGER
NCCIT	Growth Inhibition Assay			IC50=112.529 μM	SANGER
KALS-1	Growth Inhibition Assay			IC50=115.941 μM	SANGER
LB2241-RCC	Growth Inhibition Assay			IC50=116.679 μM	SANGER
HH	Growth Inhibition Assay			IC50=117.395 μM	SANGER
HD-MY-Z	Growth Inhibition Assay			IC50=118.488 μM	SANGER
EB-3	Growth Inhibition Assay			IC50=123.094 μM	SANGER
BL-70	Growth Inhibition Assay			IC50=123.127 μM	SANGER
K-562	Growth Inhibition Assay			IC50=126.245 μM	SANGER
HT-144	Growth Inhibition Assay			IC50=133.164 μM	SANGER
PF-382	Growth Inhibition Assay			IC50=134.361 μM	SANGER
RPMI-8226	Growth Inhibition Assay			IC50=135.045 μM	SANGER
NCI-H1355	Growth Inhibition Assay			IC50=135.587 μM	SANGER
LXF-289	Growth Inhibition Assay			IC50=139.781 μM	SANGER
NCI-H69	Growth Inhibition Assay			IC50=142.932 μM	SANGER
SK-MEL-1	Growth Inhibition Assay			IC50=147.13 μM	SANGER
KARPAS-299	Growth Inhibition Assay			IC50=149.12 μM	SANGER
GB-1	Growth Inhibition Assay			IC50=149.322 μM	SANGER
CMK	Growth Inhibition Assay			IC50=149.515 μM	SANGER
MPP-89	Growth Inhibition Assay			IC50=156.035 μM	SANGER
KU812	Growth Inhibition Assay			IC50=161.902 μM	SANGER
REH	Growth Inhibition Assay			IC50=162.125 μM	SANGER
NEC8	Growth Inhibition Assay			IC50=165.026 μM	SANGER
KP-N-YS	Growth Inhibition Assay			IC50=168.395 μM	SANGER
Ramos-2G6-4C10	Growth Inhibition Assay			IC50=169.915 μM	SANGER
Becker	Growth Inhibition Assay			IC50=174.18 μM	SANGER
LB647-SCLC	Growth Inhibition Assay			IC50=175.845 μM	SANGER
LU-139	Growth Inhibition Assay			IC50=178.019 μM	SANGER
QIMR-WIL	Growth Inhibition Assay			IC50=179.646 μM	SANGER
NCI-H1395	Growth Inhibition Assay			IC50=179.996 μM	SANGER
NOMO-1	Growth Inhibition Assay			IC50=182.85 μM	SANGER
GI-ME-N	Growth Inhibition Assay			IC50=187.969 μM	SANGER
KMS-12-PE	Growth Inhibition Assay			IC50=189.273 μM	SANGER
Daudi	Growth Inhibition Assay			IC50=191.128 μM	SANGER
LB996-RCC	Growth Inhibition Assay			IC50=191.699 μM	SANGER
NCI-H2107	Growth Inhibition Assay			IC50=193.739 μM	SANGER
SK-PN-DW	Growth Inhibition Assay			IC50=194.719 μM	SANGER
MC-CAR	Growth Inhibition Assay			IC50=202.253 μM	SANGER
SNB75	Growth Inhibition Assay			IC50=221.94 μM	SANGER
ES4	Growth Inhibition Assay			IC50=223.783 μM	SANGER
KARPAS-422	Growth Inhibition Assay			IC50=228.352 μM	SANGER
NCI-H1648	Growth Inhibition Assay			IC50=229.489 μM	SANGER
ES6	Growth Inhibition Assay			IC50=239.43 μM	SANGER
KNS-81-FD	Growth Inhibition Assay			IC50=241.197 μM	SANGER
JAR	Growth Inhibition Assay			IC50=256.225 μM	SANGER
NB1	Growth Inhibition Assay			IC50=260.516 μM	SANGER
D-336MG	Growth Inhibition Assay			IC50=260.698 μM	SANGER
BC-3	Growth Inhibition Assay			IC50=265.178 μM	SANGER
HCC2218	Growth Inhibition Assay			IC50=266.415 μM	SANGER
TE-9	Growth Inhibition Assay			IC50=266.627 μM	SANGER
LB1047-RCC	Growth Inhibition Assay			IC50=266.753 μM	SANGER
CTB-1	Growth Inhibition Assay			IC50=269.973 μM	SANGER
NB7	Growth Inhibition Assay			IC50=271 μM	SANGER
ST486	Growth Inhibition Assay			IC50=277.412 μM	SANGER
HCC1187	Growth Inhibition Assay			IC50=282.811 μM	SANGER
NCI-SNU-16	Growth Inhibition Assay			IC50=284.248 μM	SANGER
COR-L279	Growth Inhibition Assay			IC50=291.584 μM	SANGER
ES8	Growth Inhibition Assay			IC50=294.182 μM	SANGER
U-698-M	Growth Inhibition Assay			IC50=298.243 μM	SANGER
HEL	Growth Inhibition Assay			IC50=309.149 μM	SANGER
KINGS-1	Growth Inhibition Assay			IC50=310.674 μM	SANGER
KY821	Growth Inhibition Assay			IC50=336.595 μM	SANGER
MZ1-PC	Growth Inhibition Assay			IC50=345.618 μM	SANGER
LS-411N	Growth Inhibition Assay			IC50=354.66 μM	SANGER
SIG-M5	Growth Inhibition Assay			IC50=359.782 μM	SANGER
HT	Growth Inhibition Assay			IC50=367.711 μM	SANGER
HC-1	Growth Inhibition Assay			IC50=367.787 μM	SANGER
NCI-H1694	Growth Inhibition Assay			IC50=372.934 μM	SANGER
BB65-RCC	Growth Inhibition Assay			IC50=376.245 μM	SANGER
HAL-01	Growth Inhibition Assay			IC50=379.838 μM	SANGER
ARH-77	Growth Inhibition Assay			IC50=394.008 μM	SANGER
MZ7-mel	Growth Inhibition Assay			IC50=397.233 μM	SANGER
SIMA	Growth Inhibition Assay			IC50=403.933 μM	SANGER
DG-75	Growth Inhibition Assay			IC50=415.698 μM	SANGER
HUTU-80	Growth Inhibition Assay			IC50=419.185 μM	SANGER
KNS-42	Growth Inhibition Assay			IC50=425.815 μM	SANGER
SH-4	Growth Inhibition Assay			IC50=427.565 μM	SANGER
L-540	Growth Inhibition Assay			IC50=431.031 μM	SANGER
NB10	Growth Inhibition Assay			IC50=441.234 μM	SANGER
ES1	Growth Inhibition Assay			IC50=452.753 μM	SANGER
KMOE-2	Growth Inhibition Assay			IC50=456.711 μM	SANGER
MC116	Growth Inhibition Assay			IC50=458.116 μM	SANGER
RCC10RGB	Growth Inhibition Assay			IC50=460.005 μM	SANGER
RL95-2	Growth Inhibition Assay			IC50=460.237 μM	SANGER
Raji	Growth Inhibition Assay			IC50=468.143 μM	SANGER
CAS-1	Growth Inhibition Assay			IC50=472.073 μM	SANGER
Calu-6	Growth Inhibition Assay			IC50=475.265 μM	SANGER
KG-1	Growth Inhibition Assay			IC50=478.44 μM	SANGER
LB771-HNC	Growth Inhibition Assay			IC50=482.232 μM	SANGER
ACN	Growth Inhibition Assay			IC50=493.599 μM	SANGER
KM12	Growth Inhibition Assay			IC50=496.589 μM	SANGER
U2OS	Function assay			Binding affinity to wild type Smo expressed in U2OS cells by scintillation counting, Kd = 0.0124 μM.	23063522
U2OS	Function assay		2 hrs	Displacement of [3H]cyclopamine from wild type Smo expressed in U2OS cells after 2 hrs by scintillation counting, Ki = 0.0127 μM.	23063522
TM3	Function assay		48 hrs	Inhibition of Hedgehog signaling pathway in mouse TM3 cells bearing pTA-8xGli-Luc reporter construct assessed as transcriptional modulation of Gli after 48 hrs by luciferase assay, IC50 = 0.046 μM.	19091559
HCC827	Function assay			Displacement of [3H]-cyclopamine from SMO V404M mutant in gefitinib resistant human HCC827 cells by scintillation counting, Ki = 0.051 μM.	28787156
HEK293	Function assay		2 hrs	Displacement of BODIPY-labelled cyclopamine from human Smo receptor expressed in HEK293 cells after 2 hrs by fluorescence microscopy, IC50 = 0.064 μM.	22268551
Shh Light2	Function assay		30 hrs	Inhibition of hedgehog signaling pathway in mouse Shh Light2 cells assessed as inhibition of sonic hedgehog-induced GLI1-mediated transcriptional activity measured after 30 hrs by dual luciferase reporter gene assay, IC50 = 0.0741 μM.	27567371
U2OS	Function assay			Binding affinity to Smo D473H mutant expressed in U2OS cells by scintillation counting, Kd = 0.116 μM.	23063522
DaOY	Function assay		48 hrs	Inhibition of Hedgehog signaling in human DaOY cells assessed as downregulation of Gli1 mRNA expression after 48 hrs by RT-PCR analysis, IC50 = 0.16 μM.	24900716
U2OS	Function assay		2 hrs	Displacement of [3H]cyclopamine from Smo D473H mutant expressed in U2OS cells after 2 hrs by scintillation counting, Ki = 0.232 μM.	23063522
CHO	Function assay			Antagonist activity at human Smo receptor expressed in CHO cells by [3H]Hh-Ag binding assay, IC50 = 0.28 μM.	19091559
Shh Light2	Function assay		40 hrs	Inhibition of SHH pathway in mouse Shh Light2 cells after 40 hrs by Gli-dependent luciferase reporter gene assay, IC50 = 0.3 μM.	21592788
Shh-light2	Function assay			Inhibition of Smo-mediated Hh signaling in human Shh-light2 cells by luciferase reporter gene assay, IC50 = 0.3 μM.	22268551
M210B4	Function assay		24 hrs	Inhibition of Hedgehog signaling in mouse M210B4 cells assessed as downregulation of Ptch mRNA expression after 24 hrs by RT-PCR analysis, IC50 = 0.43 μM.	24900716
Shh-Light 2	Function assay		2 days	Antagonist activity at Smo in mouse Shh-Light 2 cells assessed as inhibition of Shh-induced Gli1-reporter activity after 2 days by dual-luciferase reporter gene method, IC50 = 0.484 μM.	23063522
Shh Light2	Function assay			Inhibition of SHH in mouse Shh Light2 cells by GLI-responsive firefly luciferase reporter gene assay, EC50 = 0.5 μM.	19309080
C3H10T1/2	Function assay			Inhibition of N-terminal SHH activated pathway in mouse C3H10T1/2 cells assessed as SAG-induced cell differentiation by alkaline phosphatase assay, IC50 = 0.6 μM.	21592788
C3H10T1/2	Function assay		6 hrs	Inhibition of SAG-induced differentiation of mouse mesenchymal pluripotent C3H10T1/2 cells to alkaline phosphatase positive oeseoblasts after 6 hrs, IC50 = 0.62 μM.	22268551
ASZ001	Function assay		48 hrs	Inhibition of Hedgehog signaling in mouse ASZ001 cells assessed as downregulation of Gli1 mRNA expression after 48 hrs by RT-PCR analysis, IC50 = 0.66 μM.	24900716
M210B4	Function assay		24 hrs	Inhibition of Hedgehog signaling in mouse M210B4 cells assessed as downregulation of Gli1 mRNA expression after 24 hrs by RT-PCR analysis, IC50 = 0.8 μM.	24900716
medulloblastoma cells	Antiproliferative assay			Antiproliferative activity against mouse medulloblastoma cells harboring heterozygous ptch1 gene by MTT assay, EC50 = 1 μM.	17417631
CHO	Function assay			Antagonist activity at mouse Smo receptor expressed in CHO cells by [3H]Hh-Ag binding assay, IC50 = 1.2 μM.	19091559
Shh-Light2	Function assay		48 hrs	Inhibition of SHH in mouse Shh-Light2 cells after 48 hrs by Gli1 reporter gene assay in presence of SAG, IC50 = 1.312 μM.	19541490
MEF	Function assay			Inhibition of Smo in mouse Ptch-deficient MEF cells assessed as inhibition of Shh-induced Gli1 transcriptional activity, IC50 = 1.5 μM.	23074541
MEF	Function assay		30 hrs	Inhibition of Smo in mouse Ptch-deficient MEF cells assessed as inhibition of Shh-induced Gli-responsive betagalactosidase activity after 30 hrs by BetaGLo assay, IC50 = 1.9 μM.	23074541
neural precursor cells	Antiproliferative assay			Antiproliferative activity against mouse neural precursor cells by colony formation assay, EC50 = 13.44 μM.	17417631
U87	Cytotoxicity assay			Cytotoxicity against human U87 cells assessed as viability in presence of beta glucuronidase, IC50 = 15.5 μM.	20116904
U87MG	Antiproliferative assay		72 hrs	Antiproliferative activity against human U87MG cells after 72 hrs by MTS assay, IC50 = 22.5 μM.	22226657
A549	Anticancer assay			Anticancer activity against human A549 cells by MTS assay, IC50 = 49 μM.	18221872
DU145	Growth inhibition assay	5 uM	96 hrs	Growth inhibition of human DU145 cells at 5 uM after 96 hrs	18249125
DU145	Growth inhibition assay	10 uM	96 hrs	Growth inhibition of human DU145 cells at 10 uM after 96 hrs	18249125
HEK293	Function assay			Inhibition of beta galactosidase in HEK293 cells	17494766
22Rv	Function assay			Reduction of expression of PTCH mRNA in human 22Rv cells	17494766
PANC1	Function assay	0.2 uM	24 hrs	Inhibition of Hh/GLI1-mediated PTCH mRNA expression in human PANC1 cells at 0.2 uM after 24 hrs by RT-PCR	20450170
PANC1	Function assay	0.4 uM	24 hrs	Inhibition of Hh/GLI1-mediated PTCH mRNA expression in human PANC1 cells at 0.4 uM after 24 hrs by RT-PCR	20450170
Shh Light2	Function assay	6.25 uM	30 hrs	Inhibition of N-palmitoylated Shh in mouse Shh Light2 cells at 6.25 uM after 30 hrs by firefly luciferase reporter gene assay	19151731
U87MG	Function assay	10 uM	4 hrs	Inhibition of Hedgehog signaling pathway in human U87MG cells assessed as down regulation of Gli1 at 10 uM after 4 hrs by RT-PCR analysis	22226657
HEK293	Function assay	5 uM	10 hrs	Displacement of BODIPY-cyclopamine from human Smo expressed in HEK293 cells at 5 uM measured after 10 hrs by DAPI staining based fluorescence microscopic assay	27736063
HEK293	Function assay	5 uM	10 hrs	Displacement of BODIPY-cyclopamine from human Smo expressed in HEK293 cells at 5 uM measured after 10 hrs by FACS analysis	27736063
C3H10T1/2	Function assay	55.5 to 4500 nM		Competitive inhibition of Smo in mouse C3H10T1/2 cells assessed as inhibition of SAG-induced Gli1 transcriptional activity at 55.5 to 4500 nM by qPCR analysis	23074541
U2OS	Function assay	5 uM	6 hrs	Antagonist activity at chimeric Smo 633 mutant expressed in U2OS cells coexpressing beta arrestin2-GFP assessed as inhibition of intracellular beta arrestin2-GFP aggregate formation at 5 uM after 6 hrs by confocal microscopy	23063522
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Chemische informatie, Opslag en Stabiliteit

Moleculair gewicht	411.62	Formule	C₂₇H₄₁NO₂	Opslag (Vanaf de ontvangstdatum)	3 jaar-20°Cpoeder
CAS-nr.	4449-51-8	SDF downloaden		Opslag van stamoplossingen	1 jaar-80°Cin oplosmiddel 1 maand-20°Cin oplosmiddel
Synoniemen	11-deoxojervine			Smiles	CC1CC2C(C(C3(O2)CCC4C5CC=C6CC(CCC6(C5CC4=C3C)C)O)C)NC1

Oplosbaarheid

In vitro
Batch:

Ethanol : 28 mg/mL

DMSO : Insoluble
(Met vocht verontreinigde DMSO kan de oplosbaarheid verminderen. Gebruik verse, watervrije DMSO.)

Water : Insoluble

Molariteitscalculator

Massa	Concentratie	Volume	Moleculair gewicht
=	×	×

Verdunningscalculator Moleculair gewicht calculator

In vivo Batch:
Clear solution	5%Ethanol 1 40%PEG300 2 5%Tween80 3 50%ddH2O Gevalideerd door Selleck labs. Mocht u aanpassingen aan deze formulering nodig hebben, neem dan contact op met ons verkoopteam voor aangepaste tests.	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.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.

In vivo Formuleringscalculator (Heldere oplossing)

Stap 1: Voer de onderstaande informatie in (Aanbevolen: Een extra dier voor het geval van verlies tijdens het experiment)

Dosering: mg/kg Gemiddeld gewicht van dieren: g Doseervolume per dier:μL Aantal dieren:

Stap 2: Voer de in vivo formulering in (Dit is alleen de calculator, geen formulering. Neem eerst contact met ons op als er geen in vivo formulering is in het gedeelte Oplosbaarheid.)

% DMSO + % + % Tween 80 + % ddH₂O

%DMSO+ %

Berekeningsresultaten:

Werkconcentratie: mg/ml;

Methode voor het bereiden van DMSO-mastervloeistof: mg geneesmiddel vooraf opgelost in μL DMSO ( Concentratie mastervloeistof mg/mL, Neem eerst contact met ons op als de concentratie de DMSO-oplosbaarheid van de partij geneesmiddel overschrijdt. )

Methode voor het bereiden van in vivo formulering: Neem μL DMSO mastervloeistof, voeg vervolgens toeμL PEG300, mengen en helder maken, voeg vervolgens toeμL Tween 80, mengen en helder maken, voeg vervolgens toe μL ddH₂O, mengen en helder maken.

Methode voor het bereiden van in vivo formulering: Neem μL DMSO mastervloeistof, voeg vervolgens toe μL Maïsolie, mengen en helder maken.

Opmerking: 1. Zorg ervoor dat de vloeistof helder is voordat u het volgende oplosmiddel toevoegt.
2. Zorg ervoor dat u het/de oplosmiddel(en) in de juiste volgorde toevoegt. U moet ervoor zorgen dat de verkregen oplossing, bij de vorige toevoeging, een heldere oplossing is voordat u verdergaat met het toevoegen van het volgende oplosmiddel. Fysische methoden zoals vortexen, echografie of een warmwaterbad kunnen worden gebruikt om het oplossen te bevorderen.

Werkingsmechanisme

Targets/IC₅₀/Ki	Smoothened (TM3Hh12 cells) 46 nM
In vitro	Cyclopamine remt de Hedgehog-signaleringsroute met een IC50 van 46 nM en blokkeert de activiteit van de menselijke Smo-receptor die tot expressie komt in CHO-K1-cellen in een [³H]Hh-Ag-bindingsanalyse met een IC50 van 280 nM. Deze verbinding remt de activiteit van de Hedgehog-route significant dosisafhankelijk in uit de darm afkomstige tumorcellijnen die Patched (PTCH)-mRNA tot expressie brengen, en induceert een groeiremming van die tumorcellijnen met 75-95% bij een concentratie van 3 μM, maar is ineffectief tegen colonkankercellen zonder PTCH-mRNA-expressie, wat suggereert dat de effecten van deze chemische behandeling Hedgehog-routegerelateerd zijn in plaats van algemeen cytotoxisch. Door de Hedgehog-signalering te blokkeren via directe interactie met Smo, remt het (10 μM) de proliferatie van SMO^high cyclopamine-responsieve cellijnen L3.6sl en Panc 05.04 met 75-80% en verhoogt het de apoptose met 2,5- tot 3,5-voudig, zonder de BxPC3-SMO^low-cellijn te beïnvloeden. Deze behandeling vermindert significant het Snail-mRNA en verhoogt de E-cadherin-transcripten in de E3LZ10.7-cellijn. Onafhankelijk van de remming van celgroei, remt het significant het invasieve fenotype van Hedgehog-afhankelijke L3.6pl-cellen, wat een >500-voudige reductie veroorzaakt in het aantal transmigrerende cellen, maar niet dat van de Hedgehog-onafhankelijke cellijn Panc-1.
Kinase Assay	Hedgehog celtest
Kinase Assay	Deze test meet het eindstadium van de Hh-signaleringsroute, namelijk de transcriptionele modulatie van Gli, met behulp van Luciferase als uitlezing (Gli-Luc-test). Cyclopamine wordt voor de test bereid door seriële verdunning in DMSO en vervolgens toegevoegd aan lege testplaten. TM3Hh12-cellen (TM3-cellen die de Hh-responsieve reportergenconstructie pTA-8xGli-Luc bevatten) worden geresuspendeerd in F12 Ham's/DMEM (1:1) met 5% FBS en 15 mM Hepes pH 7.3, toegevoegd aan testplaten en gedurende ongeveer 30 minuten bij 37 °C in 5% CO₂ geïncubeerd met deze verbinding. Vervolgens wordt 1 nM Hh-Ag 1.5 toegevoegd aan de testplaten en geïncubeerd bij 37 °C in aanwezigheid van 5% CO₂. Na 48 uur wordt ofwel Bright-Glo ofwel MTS-reagens aan de testplaten toegevoegd en wordt de luminescentie of de absorptie bij 492 nm bepaald. De IC50-waarde, gedefinieerd als het buigpunt van de logistische curve, wordt bepaald door niet-lineaire regressie van de Gli-gestuurde luciferase-luminescentie of de absorptie van de MTS-test versus log10 (concentratie) van deze chemische stof met behulp van het R-statistische softwarepakket.
In vivo	Toediening van Cyclopamine in een dosis van 50 mg/kg/dag gedurende 22 dagen eradicateert de HUCCT1-xenografts in muizen zonder duidelijke bijwerkingen. Deze verbindingbehandeling in een dosis van 1,2 mg gedurende 7 dagen induceert significante apoptose van tumorcellen en vermindert de tumormassa met respectievelijk 50-60% in van Panc 05.04- en L3.6sl-afgeleide tumoren, maar niet in de BxPC3-SMO^low-tumoren. [3] Toediening van deze chemische stof alleen remt de tumormetastasen in xenografts van E3LZ10.7 en L3.6pl diepgaand en onderdrukt metastasen volledig in combinatie met gemcitabine.
Referenties	[1] https://pubmed.ncbi.nlm.nih.gov/19091559/ [2] https://pubmed.ncbi.nlm.nih.gov/14520411/ [3] https://pubmed.ncbi.nlm.nih.gov/14520413/ [4] https://pubmed.ncbi.nlm.nih.gov/17332349/

Toepassingen

Methoden	Biomarkers	Afbeeldingen	PMID
Western blot	Snail / E-cadherin / Slug / Vimentin Gli1 / TGF-β1 / CXCR4 NF-κB / Cyclin D1 / MMP2 / MMP9		26859575
Immunofluorescence	PCNA / β-catenin Vimentin		28747625

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Veelgestelde vragen

Vraag 1:
How to reconstitute it for in vivo use in mice?

Antwoord:
One paper dissolved this compound in DMSO, and diluted it in saline: Berman DM, et al. Nature, 2003, 425(6960), 846-851. Alternatively, you can try this vehicle: 10% DMSO+30% PEG 300+5% Tween 80+ddH2O for P.O. When preparing the solution, please dissolve it in DMSO clearly first. Then add PEG300 and Tween, after they mixed well, dilute with water.

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