title: Proteomic searches comparing two (R)-lacosamide affinity baits: An electrophilic arylisothiocyanate and a photoactivated arylazide group.
abstract: We have advanced a novel strategy to search for lacosamide ((R)-1) targets in the brain proteome where protein binding is expected to be modest. Our approach used lacosamide agents containing affinity bait (AB) and chemical reporter (CR) units. The affinity bait moiety is designed to irreversibly react with the target, and the CR group permits protein detection and capture. In this study, we report the preparation and evaluation of (R)-N-(4-azido)benzyl 2-acetamido-3-(prop-2-ynyloxy)propionamide ((R)-3) and show that this compound exhibits potent anticonvulsant activities in the MES seizure model in rodents. We compared the utility of (R)-3 with its isostere, (R)-N-(4-isothiocyanato)benzyl 2-acetamido-3-(prop-2-ynyloxy)propionamide ((R)-2), in proteomic studies designed to identify potential (R)-1 targets. We showed that despite the two-fold improved anticonvulsant activity of (R)-3 compared with (R)-2, (R)-2 was superior in revealing potential binding targets in the mouse brain soluble proteome. The difference in these agents utility has been attributed to the reactivity of the affinity baits (i.e., (R)-2: aryl isothiocyanate moiety; (R)-3: photoactivated aryl azide intermediates) in the irreversible protein modification step, and we conclude that this factor is a critical determinant of successful target detection where ligand (drug) binding is modest. The utility of (R)-2 and (R)-3 in in situ proteome studies is explored.
authors.affiliation: Division of Medicinal Chemistry and Natural Products, UNC Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, North Carolina 27599-7568, USA.
authors.forename: Ki Duk
authors.initials: KD
authors.lastname: Park
authors.forename: James P
authors.initials: JP
authors.lastname: Stables
authors.forename: Rihe
authors.initials: R
authors.lastname: Liu
authors.forename: Harold
authors.initials: H
authors.lastname: Kohn
mesh_terms: D000081:Acetamides / Q000737:chemistry*; D000818:Animals; D000927:Anticonvulsants / Q000737:chemistry* / Q000494:pharmacology; D001386:Azides / Q000737:chemistry* / Q000494:pharmacology; D001921:Brain / Q000378:metabolism; D002478:Cells, Cultured; D017879:Isothiocyanates / Q000737:chemistry*; D000078334:Lacosamide; D008024:Ligands; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D020543:Proteome / Q000378:metabolism*; D012640:Seizures / Q000188:drug therapy / Q000378:metabolism; D012694:Serine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D013237:Stereoisomerism; D013329:Structure-Activity Relationship
chemical_list: D000081:Acetamides; D000927:Anticonvulsants; D001386:Azides; D017879:Isothiocyanates; D008024:Ligands; C552684:N-(4-azido)benzyl 2-acetamido-3-(prop-2-ynyloxy)propionamide; D020543:Proteome; C037152:isothiocyanic acid; D012694:Serine; D000078334:Lacosamide