title: [3H]L-657,743 (MK-912): a new, high affinity, selective radioligand for brain alpha 2-adrenoceptors.
abstract: L-657,743 (MK-912), a highly potent and selective alpha 2-adrenoceptor antagonist was tritiated to a high specific activity and its binding characteristics to brain tissue were determined. The specific binding of [3H]L-657,743 to rat cerebrocortex was saturable, reversible, and dependent on tissue concentration. In saturation studies, [3H]L-657,743 binding was resolved into two high affinity components exhibiting Kd values of 86 pM and 830 pM with densities of 82 fmol/mg protein and 660 fmol/mg protein, respectively. Based on the binding potencies of a variety of compounds with differing receptor selectivities, the sites labeled by [3H]L-657,743 were characteristic of alpha 2-adrenoceptors. In contrast to alpha 2-antagonists, alpha 2-agonists displayed shallow competition curves. In the presence of 100 microM GTP, Gpp(NH)p or 150 mM NaCl, the competition curve for epinephrine was shifted to the right, whereas that for yohimbine was unaffected. In studies utilizing human cerebrocortical tissue, [3H]L-657,743 also bound with high affinity to sites characteristic of alpha 2-adrenoceptors.
authors.affiliation: Department of Microbial Pharmacometrics, Merck, Sharp and Dohme Research Laboratories, West Point, PA 19486.
authors.forename: D J
authors.initials: DJ
authors.lastname: Pettibone
authors.forename: S D
authors.initials: SD
authors.lastname: Flagg
authors.forename: J A
authors.initials: JA
authors.lastname: Totaro
authors.forename: B V
authors.initials: BV
authors.lastname: Clineschmidt
authors.forename: J R
authors.initials: JR
authors.lastname: Huff
authors.forename: S D
authors.initials: SD
authors.lastname: Young
authors.forename: R
authors.initials: R
authors.lastname: Chen
mesh_terms: D000317:Adrenergic alpha-Antagonists / Q000378:metabolism*; D000818:Animals; D002540:Cerebral Cortex / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D011807:Quinolizines / Q000378:metabolism*; D011869:Radioligand Assay; D051381:Rats; D011919:Rats, Inbred Strains; D011942:Receptors, Adrenergic, alpha / Q000378:metabolism*; D015016:Yohimbine / Q000378:metabolism
chemical_list: D000317:Adrenergic alpha-Antagonists; D011807:Quinolizines; D011942:Receptors, Adrenergic, alpha; C054290:L 657743; D015016:Yohimbine