dermorphin and morphiceptin

Topics: Amino Acid Sequence; Analgesics; Analgesics, Opioid; Animals; Endorphins; Humans; Molecular Sequence Data; Oligopeptides; Opioid Peptides

We examined the in vitro and in vivo bioactivities of several families of peptidomimetic opioids including: constrained linear enkephalin (n = 12 analogs), dermorphin (n = 9 analogs) and morphiceptin (n = 17 analogs). The biological activities were assessed in vitro by examining the inhibitory effects of these agents on the electrically evoked contractions of the guinea pig ileum (GPI) and the mouse vas deferens (MVD) preparations. The in vivo bioactivities were determined from the antinociceptive activity of these agents on the 52.5 degrees C hot-plate test after spinal administration of rats with chronically placed spinal catheters. Examination of the effect of cyclization, incorporation of retro-inverso bonds and substitutions of D- or constrained amino acids reveals systematic changes in the activity of these agents. There was a significant correlation between the potency of these agents in the hot-plate bioassay and their activity in the GPI and, to a lesser extent, in the MVD tests. Examination of the ability of naltrindole (a delta selective antagonist) to reverse the drug action and the respective potency on the GPI and MVD, showed that a correlation exists with actions on the MVD, but not on the GPI, consistent with the likelihood that agents with high MVD/GPI ratios in vitro act at the mu sites, whereas those with low MVD/GPI ratios act at the delta receptor in the spinal cord. The close correlations between activity in the GPI and spinal cord suggest that the structural requirements for potency in the smooth muscle and in the spinal cord are essentially the same as those mu receptors that mediate nociceptive transmission.

Topics: Amino Acid Sequence; Analgesia; Analgesics, Opioid; Animals; Dose-Response Relationship, Drug; Endorphins; Enkephalins; Injections, Spinal; Male; Mice; Molecular Sequence Data; Muscle, Smooth; Naloxone; Naltrexone; Narcotic Antagonists; Nociceptors; Oligopeptides; Opioid Peptides; Rats; Rats, Sprague-Dawley; Receptors, Opioid, delta; Receptors, Opioid, mu; Spinal Cord; Structure-Activity Relationship; Vas Deferens

The structural basis of opioid receptors (OPRs) for the subtype-selective binding of DAMGO, a mu-opioid receptor selective ligand, was investigated using chimeric mu/kappa-OPRs. Replacement of the region from the middle of the fifth transmembrane domain to the C-terminal of mu-OPR with the corresponding region of mu-OPR remarkably decreased the binding affinity to DAMGO, while the reciprocal chimera revealed the high affinity to DAMGO. These results indicate that DAMGO distinguishes between mu- and mu-OPRs at the region around the third extracellular loop, different from the case of the distinction between mu-and delta-OPRs in which the region around the first extracellular loop is important. Furthermore, displacement studies revealed that the region around the third extracellular loop is involved in the discrimination between mu- and kappa-OPRs not only by peptidic mu- selective ligands but also by non-peptidic ligands, such as morphine and naloxone.

Topics: Amino Acid Sequence; Analgesics; Animals; Cells, Cultured; Endorphins; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalins; Ligands; Molecular Sequence Data; Morphine; Naloxone; Oligopeptides; Opioid Peptides; Receptors, Opioid; Receptors, Opioid, delta; Receptors, Opioid, kappa; Receptors, Opioid, mu; Recombinant Fusion Proteins; Somatostatin; Structure-Activity Relationship

To elucidate the topochemical requirements for bioactivities of morphiceptin (Tyr-Pro-Phe-Pro-NH2) and dermorphin (Tyr-D-Ala-Phe-Gly-Tyr-Pro-Ser-NH2), we have designed and synthesized two diastereomers Tyr-(L and D)-(NMe)Ala-Phe-D-Pro-NH2. Both the analogs display high activities in the guinea pig ileum assay. The only difference in the composition of these two diastereomers arises from the chirality at residue 2. The high mu-receptor activities are attributed to structures where the Tyr1-L-(NMe)Ala2 amide bond assumes a cis configuration while the Tyr1-D-(NMe)Ala2 amide bond assumes a trans configuration. Accessible space studied for the second residues of these molecules confirms the fact that the L-(NMe)Ala2 analog belongs to the morphiceptin family of opioids while the D-(NMe)Ala2 analog belongs to the dermorphin class of opioids. The similarity in the spacial array of the analogs explains their high mu-receptor activities and indicates that they are likely recognized at the same opioid receptor.

Topics: Amino Acid Sequence; Animals; Endorphins; Guinea Pigs; Ileum; Male; Mice; Molecular Conformation; Molecular Sequence Data; Oligopeptides; Opioid Peptides; Receptors, Opioid; Receptors, Opioid, delta; Receptors, Opioid, mu; Stereoisomerism; Structure-Activity Relationship; Vas Deferens