buprenorphine and naltrindole

δ-Opioid receptor (

Topics: Animals; Behavior, Animal; Benzamides; Buprenorphine; Drug Partial Agonism; Mice; Naltrexone; Piperazines; Receptors, Opioid, delta

Buprenorphine is widely used as an analgesic drug and it is also increasingly considered for maintenance and detoxification of heroin addicts. It is a potent micro -receptor partial agonist with a long duration of action. An inverted U-shaped dose-effect curve for buprenorphine conditioned place preference (CPP) has been shown previously.. We re-evaluated the CPP effects of buprenorphine by taking into account the particular kinetic properties of the drug in the design of the experiments.. An unbiased CPP procedure with different wash-out periods was used to investigate a possible influence of the long duration of action of buprenorphine on the outcome of the experiment.. Following a standard procedure (drug and vehicle conditioning on alternating days), the inverted U-shaped dose-effect curve was reproduced (no CPP at 0.01 mg/kg, significant CPP at 0.1 and 1.0 mg/kg, and no CPP at 3.16 and 10 mg/kg, IP). However, when there was a 48 h interval between drug and vehicle conditioning, there was a clear tendency towards CPP for the two highest doses, and when there was a 72-h interval between drug and vehicle conditioning, significant CPP was seen. Naloxone (0.215 mg/kg SC), haloperidol (0.215 mg/kg IP) and U-50488 (1.0 mg/kg SC) blocked buprenorphine (1.0 mg/kg) CPP. Buprenorphine CPP was also blocked by coadministration of naltrindole (3.16 mg/kg IP), nor-binaltorphimine (4.64 mg/kg SC), and naloxonebenzoylhydrazone (0.464 mg/kg SC). However, the data suggest that blockade by the three latter drugs was due to state-dependency effects. Buprenorphine at doses of 1.0 mg/kg and higher also produced locomotor sensitization across the 3 drug conditioning days. The sensitization produced by 1.0 mg/kg buprenorphine was blocked by haloperidol and U-50488, but not by naloxone, naltrindole, nor-binaltorphimine, and naloxonebenzoylhydrazone.. The present results suggest that the reported lack of CPP effects at high doses of buprenorphine may be due to factors in the experimental design, resulting in a carry-over effect from drug- to vehicle conditioning. They also suggest that buprenorphine, like other opiates, produces its CPP effects via micro -receptors, although kappa-antagonistic mechanisms also appear to be involved. The implications of these findings for the safety of buprenorphine for human use are discussed.

Topics: Animals; Buprenorphine; Conditioning, Operant; Dose-Response Relationship, Drug; Male; Motor Activity; Naloxone; Naltrexone; Narcotic Antagonists; Narcotics; Nociceptin Receptor; Rats; Rats, Sprague-Dawley; Receptors, Opioid; Receptors, Opioid, kappa; Time Factors

N-Cyclopropylmethyl-[7alpha,8alpha,2', 3']-cyclohexano-1'[S]-hydroxy-6,14-endo-ethenotetrahydronororip avine (BU48) is a novel, ring-constrained analog of buprenorphine. In vivo, BU48 (0.1-10 mg/kg s.c.) produced brief, nonlethal convulsions in mice followed by brief Straub tail and a short period of catalepsy characteristic of BW373U86 and other nonpeptidic delta-receptor agonists. BU48-induced convulsions were sensitive to antagonism by naltrindole (10 mg/kg s.c.) and were also prevented by administration of the putative delta(1) antagonist 7-benzylidenenaltrexone and the putative delta(2) antagonist naltriben, with the latter being more potent. In the abdominal stretch assay in the mouse, only low-efficacy antinociceptive activity of BU48 (0.1-10 mg/kg) was seen. This was reversed by the kappa-opioid antagonist norbinaltorphimine (32 mg/kg s.c.) but not by the delta-opioid antagonist naltrindole (10 mg/kg s.c.). BU48 (10 mg/kg s.c.) acted as a delta-antagonist in this assay. In mouse brain homogenates, BU48 had high (nanomolar) binding affinity for all three opioid receptors in the order mu > delta = kappa. In vitro, the compound acted as a potent (EC(50) = 1.4 nM) kappa-opioid agonist in the guinea pig ileum and a potent (EC(50) = 0.2 nM) delta-opioid agonist in the mouse vas deferens but showed partial agonist activity at the rat cloned delta-opioid (40%) and human cloned kappa-opioid (59%) receptors with very low efficacy at the rat cloned mu-opioid receptor (10%); findings consistent with its in vivo profile. BU48 is the first described compound that produces delta-opioid-mediated convulsions without any evidence of delta-opioid-mediated antinociception and will be a useful tool in investigations of the delta-opioid receptor.

Topics: Analgesics, Opioid; Animals; Benzamides; Brain; Buprenorphine; Convulsants; Electric Stimulation; Guanosine 5'-O-(3-Thiotriphosphate); Guinea Pigs; Ileum; In Vitro Techniques; Male; Mice; Muscle Contraction; Muscle, Smooth; Naltrexone; Narcotic Antagonists; Pain Measurement; Piperazines; Radioligand Assay; Rats; Receptors, Opioid, delta; Receptors, Opioid, kappa; Seizures; Vas Deferens

An aspartic acid at residue 95 (Asp95) in the delta receptor has previously been shown to be critical for the binding affinity of selective delta agonists. To gain a better understanding of the functional consequence of agonist action at the delta receptor, the Asp95 residue was mutated to an asparagine (D95N) and opioids were tested for binding and functional activation of the wild-type and mutant delta receptors. Selective agonists such as [D-Ser2,D-Leu5]enkephalin-Thr6 (DSLET) and [D-Ala2,D-Leu5]enkephalin (DADLE) had greatly reduced affinity for the D95N mutant receptor but still inhibited cAMP accumulation, which indicated that the mutant receptor was still functionally coupled to adenylyl cyclase. Antagonist binding was not affected by the Asp95 mutation. Similarly, the partial agonist buprenorphine bound with equally high affinity to the D95N mutant and the wild-type delta receptor, which indicated that Asp95 is not essential for the binding affinity of this opioid. Buprenorphine did not affect cAMP accumulation in HEK 293 cells expressing the D95N mutant, and it blocked the ability of DSLET and bremazocine to inhibit cAMP accumulation via the D95N mutant, which indicated that buprenorphine acts as an antagonist at the D95N mutant. These findings confirm the essential role of Asp95 in the activation of the delta receptor by agonists and reveal a molecular basis of the unique property of buprenorphine.

Topics: Analgesics, Opioid; Animals; Buprenorphine; Cells, Cultured; Cyclic AMP; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Mice; Mutagenesis; Naltrexone; Narcotic Antagonists; Receptors, Opioid, delta