title: Characterization of peripheral human cannabinoid receptor (hCB2) expression and pharmacology using a novel radioligand, [35S]Sch225336.
abstract: Studies to characterize the endogenous expression and pharmacology of peripheral human cannabinoid receptor (hCB2) have been hampered by the dearth of authentic anti-hCB2 antibodies and the lack of radioligands with CB2 selectivity. We recently described a novel CB2 inverse agonist, N-[1(S)-[4-[[4-methoxy-2-[(4methoxyphenyl)sulfonyl] phenyl]sulfonyl] phenyl]ethyl]methane-sulfonamide (Sch225336), that binds hCB2 with high affinity and excellent selectivity versus hCB1. The precursor primary amine of Sch225336 was prepared and reacted directly with [(35)S]mesyl chloride (synthesized from commercially obtained [(35)S]methane sulfonic acid) to generate [(35)S]Sch225336. [(35)S]Sch225336 has high specific activity (>1,400 Ci/mmol) and affinity for hCB2 (65 pm). Using [(35)S]Sch225336, we assayed hemopoietic cells and cell lines to quantitate the expression and pharmacology of hCB2. Lastly, we used [(35)S]Sch225336 for detailed autoradiographic analysis of CB2 in lymphoid tissues. Based on these data, we conclude that [(35)S]Sch225336 represents a unique radioligand for the study of CB2 endogenously expressed in blood cells and tissues.
authors.affiliation: Department of Inflammation, Schering-Plough Research Institute, Kenilworth, New Jersey 07033, USA.
authors.forename: Waldemar
authors.initials: W
authors.lastname: Gonsiorek
authors.forename: David
authors.initials: D
authors.lastname: Hesk
authors.forename: Shu-Cheng
authors.initials: SC
authors.lastname: Chen
authors.forename: David
authors.initials: D
authors.lastname: Kinsley
authors.forename: Jay S
authors.initials: JS
authors.lastname: Fine
authors.forename: James V
authors.initials: JV
authors.lastname: Jackson
authors.forename: Loretta A
authors.initials: LA
authors.lastname: Bober
authors.forename: Gregory
authors.initials: G
authors.lastname: Deno
authors.forename: Hong
authors.initials: H
authors.lastname: Bian
authors.forename: James
authors.initials: J
authors.lastname: Fossetta
authors.forename: Charles A
authors.initials: CA
authors.lastname: Lunn
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Kozlowski
authors.forename: Brian
authors.initials: B
authors.lastname: Lavey
authors.forename: John
authors.initials: J
authors.lastname: Piwinski
authors.forename: Satwant K
authors.initials: SK
authors.lastname: Narula
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Lundell
authors.forename: R William
authors.initials: RW
authors.lastname: Hipkin
mesh_terms: D000818:Animals; D001345:Autoradiography; D015153:Blotting, Western; D016466:CHO Cells; D006224:Cricetinae; D018922:HL-60 Cells; D006801:Humans; D008214:Lymphocytes / Q000737:chemistry; D011869:Radioligand Assay; D043885:Receptor, Cannabinoid, CB2 / Q000819:agonists* / Q000032:analysis*; D013154:Spleen / Q000737:chemistry; D013449:Sulfonamides / Q000378:metabolism*
chemical_list: D043885:Receptor, Cannabinoid, CB2; C514445:Sch225336; D013449:Sulfonamides