Capadenoson Inhibitor

Solid. Powder

98.00%

Store at -20 degree C for 3 years powder
-80 degree C for 2 years in solvent

520.03

DMSO: 50mg/mL (96.15mM)
Water: Insoluble
(<1mg/ml refers to the product slightly soluble or insoluble)

C25H18ClN5O2S2

Nc(nc(c(C#N)c1-c(cc2)ccc2OCCO)SCc2csc(-c(cc3)ccc3Cl)n2)c1C#N

544417-40-5

Membranes from the human cortex are prepared. [35S]GTPgammaS binding is measured. Briefly, 5ug of membrane protein is incubated in a total volume of 160uL for 2 hr at 25 degree C in a shaking water bathours. [35S]GTPgammaS binding in control incubations and in the presence of Capadenoson showed a linear time course up to this incubation time. Binding buffer contained 50mM Tris/HCl, pH 7.4, 2mM triethanolamine, 1mM EDTA, 5mM MgCl2, 10uM GDP, 1mM dithiothreitol, 100mM NaCl, 0.2units/mL adenosine deaminase, 0.2nM [35S]GTPgammaS, and 0.5% bovine serum albuminutes. Non-specific binding is determined in the presence of 10uM GTPgammaS. Incubations are terminated through filtration of the samples over multiscreen FB glass fiber filters followed by two washes with binding buffer. The filters are dried, coated with scintillator and counted for radioactivity. Binding curves of [35S]GTPgammaS are analyzed by nonlinear regression using GraphPad Prism.

To further elucidate the pharmacological properties of Capadenson, GTP shift assays are performed with the standard full A1-agonist CCPA and the A1-antagonist DPCPX. CCPA shows a Ki value of 4.2nM in the binding assay on rat cortical brain membranes. In the presence of 1mM GTP this Ki value shifts to a value of 64nM. Therefore the GTP shift for CCPA is 15. DPCPX shows a GTP shift of 1 with virtually identical Ki values in the absence and presence of GTP. Capadenson shows a Ki value of 24nM in the binding assay. In the presence of 1mM GTP this Ki value shifts to a value of 116nM resulting in a GTP shift of 5 for Capadenoson.

GPCR/G Protein; Adenosine Receptor