Synthesis and pharmacological characterization of [(125)I]MRS5127, a high affinity, selective agonist radioligand for the A3 adenosine receptor. Biochem Pharmacol 2010 Apr 01;79(7):967-73
Date
11/18/2009Pubmed ID
19917269Pubmed Central ID
PMC2815200DOI
10.1016/j.bcp.2009.11.009Scopus ID
2-s2.0-77449155693 (requires institutional sign-in at Scopus site) 21 CitationsAbstract
A recently reported selective agonist of the human A(3) adenosine receptor (hA(3)AR), MRS5127 (1'R,2'R,3'S,4'R,5'S)-4'-[2-chloro-6-(3-iodobenzylamino)-purine]-2',3'-O-dihydroxy-bicyclo-[3.1.0]hexane, was radioiodinated and characterized pharmacologically. It contains a rigid bicyclic ring system in place of a 5'-truncated ribose moiety, and was selected for radiolabeling due to its nanomolar binding affinity at both human and rat A(3)ARs. The radioiodination of the N(6)-3-iodobenzyl substituent by iododestannylation of a 3-(trimethylstannyl)benzyl precursor was achieved in 73% yield, measured after purification by HPLC. [(125)I]MRS5127 bound to the human A(3)AR expressed in membranes of stably transfected HEK 293 cells. Specific binding was saturable, competitive, and followed a one-site binding model, with a K(d) value of 5.74+/-0.97nM. At a concentration equivalent to its K(d), non-specific binding comprised 27+/-2% of total binding. In kinetic studies, [(125)I]MRS5127 rapidly associated with the hA(3)AR (t(1/2)=0.514+/-0.014min), and the affinity calculated from association and dissociation rate constants was 3.50+/-1.46nM. The pharmacological profile of ligands in competition experiments with [(125)I]MRS5127 was consistent with the known structure-activity-relationship profile of the hA(3)AR. [(125)I]MRS5127 bound with similar high affinity (K(d), nM) to recombinant A(3)ARs from mouse (4.90+/-0.77), rabbit (2.53+/-0.11), and dog (3.35+/-0.54). For all of the species tested, MRS5127 exhibited A(3)AR agonist activity based on negative coupling to cAMP production. Thus, [(125)I]MRS5127 represents a new species-independent agonist radioligand for the A(3)AR. The major advantage of [(125)I]MRS5127 compared with previously used A(3)AR radioligands is its high affinity, low degree of non-specific binding, and improved A(3)AR selectivity.
Author List
Auchampach JA, Gizewski ET, Wan TC, de Castro S, Brown GG Jr, Jacobson KAAuthors
John A. Auchampach PhD Professor in the Pharmacology and Toxicology department at Medical College of WisconsinTina C. Wan PhD Research Scientist II in the Pediatrics department at Medical College of Wisconsin
MESH terms used to index this publication - Major topics in bold
AdenosineAdenosine A3 Receptor Agonists
Animals
Cells, Cultured
Dogs
Humans
Iodine Radioisotopes
Mice
Rabbits
Radioligand Assay
Radiopharmaceuticals
Rats
Structure-Activity Relationship
