title: Inhibition of the enkephalin-metabolizing enzymes by the first systemically active mixed inhibitor prodrug RB 101 induces potent analgesic responses in mice and rats.
abstract: N-([(R,S)-2-benzyl-3[(S)(2-amino-4-methylthio)butyl dithio]-1-oxopropyl)-L-phenylalanine benzyl ester (RB101) is the first systemically active prodrug generating through a biologically dependent cleavage of the disulfide bond the potent (S)2-amino-1-mercapto-4-methylthio butane (aminopeptidase N) (IC50 = 11 nM) and N-[(R,S)-2-mercapto-methyl-1-oxo-3-phenylpropyl]-L-phenylalanine (neutral endopeptidase) (IC50 = 2 nM) inhibitors (aminopeptidase N). RB101 easily crosses the blood-brain barrier, as shown by the observed complete inhibition of cerebral endopeptidase 24.11 after i.v. injection in mice. The prodrug induces strong, dose-dependent antinociceptive responses in mice after i.v., i.p. or s.c. administration, in the hot plate (ED50 = 9 mg/kg) and phenylbenzoquinone-induced writhing (ED50-3.25 mg/kg) tests in mice, which are currently used in analgesics screening. RB101 is also active in the tail-flick and tail-electric stimulation tests in rats. In contrast, under disulfide forms, the above selective aminopeptidase N or endopeptidase 24.11 inhibitors are inactive after i.v. administration and their association 3 times less potent than RB101 alone. In all the tests used, the pain-alleviating effect of RB101 was suppressed by naloxone, but, except for the tail-flick and the motor response to tail-electric stimulation, not by the delta-selective antagonist naltrindole. The preferential involvement of mu opioid receptors in the analgesic effects of endogenous enkephalins, whose extracellular levels are increased by the two RB101-generated inhibitors, is suggested by the similar apparent pA2 values for RB101-naloxone (pA2: 7.53 +/- 0.046) and DAMGO (mu-selective ligand)-naloxone (pA2: 7.38 +/- 0.049).(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Départment de Chimie Organique, U 266 INSERM, UFR des Sciences Pharmaceutiques et Biologiques, Paris, France.
authors.forename: F
authors.initials: F
authors.lastname: Noble
authors.forename: J M
authors.initials: JM
authors.lastname: Soleilhac
authors.forename: E
authors.initials: E
authors.lastname: Soroca-Lucas
authors.forename: S
authors.initials: S
authors.lastname: Turcaud
authors.forename: M C
authors.initials: MC
authors.lastname: Fournie-Zaluski
authors.forename: B P
authors.initials: BP
authors.lastname: Roques
mesh_terms: D000626:Aminopeptidases / Q000037:antagonists & inhibitors*; D000698:Analgesia; D000700:Analgesics / Q000008:administration & dosage / Q000494:pharmacology*; D000818:Animals; D002462:Cell Membrane / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D004220:Disulfides / Q000494:pharmacology*; D004347:Drug Interactions; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; D004745:Enkephalins / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000008:administration & dosage / Q000494:pharmacology; D007275:Injections, Intravenous; D007276:Injections, Intraventricular; D007279:Injections, Subcutaneous; D008297:Male; D051379:Mice; D009270:Naloxone / Q000494:pharmacology; D015260:Neprilysin / Q000037:antagonists & inhibitors*; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000494:pharmacology; D011355:Prodrugs / Q000008:administration & dosage / Q000494:pharmacology*; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D000700:Analgesics; D004220:Disulfides; D004745:Enkephalins; D004791:Enzyme Inhibitors; D011355:Prodrugs; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; C074024:RB 101; D009270:Naloxone; D010649:Phenylalanine; D000626:Aminopeptidases; D015260:Neprilysin