The adenosine receptor affinities and monoamine oxidase B inhibitory properties of sulfanylphthalimide analogues

1,3,7-Triethyl-substituted xanthines—possess nanomolar affinity for the adenosine A1 receptor

“…32 The experimental results of the radioligand binding experiments to adenosine A 1 receptors in rat whole brain membranes and 34 as radioligands for the adenosine A 1 and A 2A receptors, respectively. In addition, the adenosine A 2A receptor binding studies were determined in the presence of N 6 -cyclopentyladenosine (CPA) to minimize the binding of [ 3 H]NECA to adenosine A 1 receptors.…”

“…33 The radioligand [ 3 H]DPCPX (K d = 0.36 nM) was used at a concentration of 0.1 nM as documented in the literature. 32,33 The incubations contained 120 lg membrane protein suspension, 0.1 nM [ 3 H] DPCPX, 0.1 units/mL adenosine deaminase, with or without 0.1 mM GTP, the test compound and 1% DMSO. The addition of GTP to a final concentration of 0.1 mM was done according to literature specifications.…”

“…The assays were performed as described in literaure. 32 Nonspecific binding was defined by the addition of either 100 lM CPA 32,34 or unlabelled 10 lM DPCPX. 5 The two competitors resulted in the same nonspecific binding determination for the adenosine A 1 receptor radioligand binding assays and for the adenosine A 2A receptor radioligand binding assays.…”

“…GTP shift assay (rat whole brain membranes): The membrane preparation was performed under the same conditions as described above for the adenosine A 1 receptor radioligand binding assay (Section 4.2.1). 32,33 The protein content of all membrane preparations were determined with Bradford reagent according to the method described by Bradford. 43 GTP shift assay protocol: The GTP shift assays performed with either rat cortical or whole brain membranes were carried out in a final assay volume of 1 mL of the appropriate 50 mM Tris-HCl buffer.…”

“…On the day of the membrane preparation, the rat whole brains and rat striata were prepared according to the protocol described in literature. 32 The protein content of all membrane preparations were determined with Bradford reagent according to the method described by Bradford. 43 Radioligand binding assay protocol: For the radioligand binding assays at adenosine A 1 receptors, the incubations contained 120 lg of membrane protein suspension (rat whole brain mem- 32,34 DMSO was used to prepare all stock solutions of the test compounds.…”