C. Gillen et al., Affinity, potency and efficacy of tramadol and its metabolites at the cloned human mu-opioid receptor, N-S ARCH PH, 362(2), 2000, pp. 116-121
The present study was conducted to characterise the centrally active analge
sic drug tramadol hydrochloride [(1RS,2RS)-2-[(dimethyl-amino)-methyl]-1-(3
-methoxyphenyl)-cyclohexanol hydrochloride] and its metabolites M1, M2, M3,
M4 and M5 at the cloned human mu-opioid receptor. Membranes from stably tr
ansfected Chinese hamster ovary (CHO) cells were used to determine the four
parameters of the ligand-receptor interaction: the affinity of (+/-)-trama
dol and its metabolites was determined by competitive inhibition of [H-3]na
loxone binding under high and low salt conditions. The agonist-induced stim
ulation of [S-35]GTP gamma S binding permits the measurement of potency (EC
50), efficacy (E-max = maximal stimulation) and relative intrinsic efficacy
(effect as a function of receptor occupation). The metabolite (+)-M1 showe
d the highest affinity (K-i = 3.4 nM) to the human mu-opioid receptor, foll
owed by (+/-)-M5 (K-i = 100 nM), (-)-M1 (K-i = 240 nM) and (+)-tramadol (K-
i = 2.4 mu M). The [S-35]GTP gamma S binding assay revealed an agonistic ac
tivity for the metabolites (+)-M1, (-)-M1 and (+/-)-M5 with the following r
ank order of intrinsic efficacy: (+)-M1>(+/-)M5>(-)-M1. The metabolites (+/
-)-M2, (+/-)-M3 and (+/-)-M4 displayed only weak affinity (K-i > 10 mu M) a
nd had no stimulatory effect on GTP gamma S binding. These data indicate th
at the metabolite (+)-M1 is responsible for the gamma-opioid-derived analge
sic effect.