nalorphine and ubenimex

The tripeptide Tyr-Gly-Gly, a hydrolysis product of enkephalins and related opioid peptides obtained with 'enkephalinase', was identified and quantified in various regions of mouse brain by means of HPLC and a sensitive and specific radioimmunoassay. Similar levels i.e. about 8 pmol/brain were found after the animals were killed by various procedures, including microwave irradiation, suggesting its pre-mortem formation. The distribution of Tyr-Gly-Gly immunoreactivity among brain regions was highly heterogeneous and paralleled to a certain extent the [Met5]enkephalin distribution, molar levels of Tyr-Gly-Gly representing 10-30% of those of the enkephalin. Following gentle homogeneisation of striata in 0.32 M sucrose and centrifugation, 73% of Tyr-Gly-Gly immunoreactivity was recovered in the supernatant, a result consistent with its extracellular localisation in vivo. Administration of enkephalinase inhibitors rapidly elicited marked decrease in Tyr-Gly-Gly immunoreactivity whereas bestatin, an aminopeptidase inhibitor, elicited 100% increase and captopril, an ACE inhibitor, was without significant effect. These data indicate that the tripeptide is in a dynamic state in the brain and that its levels might reflect the release of endogenous enkephalins or related opioid peptides and their subsequent metabolism by enkephalinase.

Topics: Animals; Brain; Captopril; Chromatography, High Pressure Liquid; Enkephalin, Leucine; Enkephalin, Methionine; Leucine; Male; Mice; Microwaves; Nalorphine; Neprilysin; Oligopeptides; Protease Inhibitors; Radioimmunoassay; Thiorphan; Tiopronin

The analgesic activity of some opioid peptides which display a relative selectivity for either the mu-receptor subtype, [D-Ala2, MePhe4, Gly-ol5]enkephalin (DAGO) or the delta-receptor subtype. [D-Ala2, D-Leu5]enkephalin (DADLE), [D-Ser2, Leu5]enkephalyl-Thr (DSLET) and [D-Thr2, Leu5]enkephalyl-Thr (DTLET) is highly correlated with their affinity for central or peripheral mu- but not delta-receptors. Moreover their analgesic effects as well as those elicited by degrading enzyme inhibitors (bestatin + thiorphan) of endogenous enkephalins were easily antagonized by naloxone with similar pA2 values but not by the delta-antagonist ICI 154,129. Therefore the analgesia produced by opioid peptides including endogenous enkephalins is likely connected to mu-receptor stimulation. Finally, there was no obvious potentiation by delta-agonists of the analgesia resulting from either administration of the mu-agonist morphine or endogenous enkephalins. This suggested that in the hot plate test, there is no modulation of the effect resulting from mu-receptor stimulation by a delta-receptor interaction. Likewise, enkephalinergic activity such as that due to thiorphan + bestatin does not appear to be regulated through mu- or delta-receptor stimulation.

Topics: Analgesics; Animals; Enkephalin, Leucine; Enkephalins; Guinea Pigs; In Vitro Techniques; Leucine; Male; Mice; Morphine; Muscle Contraction; Muscle, Smooth; Nalorphine; Naloxone; Receptors, Opioid; Receptors, Opioid, delta; Receptors, Opioid, mu; Thiorphan; Tiopronin