Bms 933043

Donepezil hydrochloride (HCl) was purchased from Sigma-Aldrich. Scopolamine HCl, EVP-6124 HCl (Encenicline; (R)-7-chloro-N-(quinuclidin-3-yl)benzo[b]thiophene-2-carboxamide HCl)[22 (link)], RG3487 HCl (also known as R3487, MT-666 and MEM 3454; N-[(3S)-1-Azabicyclo[2.2.2]oct-3-yl]-1H-indazole-3-carboxamide HCl)[47 (link)] and BMS-933043 ((2R)-N-(6-(1H-imidazol-1-yl)-4-pyrimidinyl)-4'H-spiro[4-azabicyclo[2.2.2]octane-2,5'-[1 (link),3 (link)]oxazol]-2'-amine) [13 , 43 (link)] were synthesized at Bristol Myers Squibb. The chemical structures of these compounds are shown in Fig 1. Doses cited refer to the free base form of the compounds. Vehicles were sterile normal saline for scopolamine, donepezil, R3487 and EVP-6124, with sterile normal saline at pH 4.0 for BMS-933043. All drugs were administered intramuscularly (i.m.) in a dosing volume of 0.1–0.2 ml/kg. The dose range examined for each compound was as follows: Donepezil (0.03–0.3 mg/kg), BMS-933043 (0.03–1.0 mg/kg), EVP-6124 (0.003–1.0 mg/kg) and R3487 (0.03–1.0 mg/kg).

HEK293 cell lines expressing either the rat or human α7 nAChR (+ resistance to inhibitors of cholinesterase 3 (RIC-3)), rat α1β1δε nAChR, rat α4β2 nAChR, rat α3β4 nAChR or human 5-HT_3A receptor were developed by Bristol-Myers Squibb (Wallingford, CT, USA). The following chemicals were synthesized by Bristol-Myers Squibb: A-582941, BMS-933043, EVP-6124, NS-6740 (4-[[5-[3-(trifluoromethyl)phenyl]-2-furanyl]carbonyl]-1,4-diazabicyclo[3.2.2]nonane), PNU-282987 (N-(3R)-1-azabicyclo[2.2.2]oct-3-yl-4-chloro-benzamide hydrochloride), TC-5619, [³H]-A585539 (³H-2,2-dimethyl-5-(6-phenyl-3-pyridazinyl)-5-aza-2-azoniabicyclo[2.2.1]heptane iodide; specific activity 71 Ci/mmol, concentration 1mCi/ml). Suppliers for other reagents were as follows: (±)epibatidine dihydrochloride hydrate (FLIPR studies), phencyclidine hydrochloride (PCP), (+)MK-801 hydrogen maleate ([5R,10S]-[+]-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine), S(+)ketamine HCl and (-)nicotine hydrogen tartrate salt (Sigma-Aldrich, Saint Louis, Mo); methyllycaconitine citrate, (±)epibatidine (binding studies) and MDL-72222 (tropanyl 3,5-dichlorobenzoate BioTechne, Minneapolis, MN); [³H]granisetron ([³H]BRL-43694, specific activity 83.66 Ci/mmol, concentration 1 mCi/ml; Perkin Elmer, Waltham, MA).