ABSTRACT
Novel substituted amino acid tethered norsufentanil derivatives were synthesized by the four-component Ugi reaction. Norsufentanil was reacted with succinic anhydride to produce the corresponding carboxylic acid. The resulting carboxylic acid has undergone a multicomponent reaction with different aldehydes, amines, and isocyanides to produce a library of the desired compounds. In all cases, amide bond rotation was observed in the NMR spectra. In vivo analgesic activity of the synthesized compounds was evaluated by a tail flick test. Very encouraging results were obtained for a number of the synthesized products. Some of the synthesized compounds such as 5a, 5b, 5h, 5j, and 5r were found to be more potent than sufentanil, sufentanil citrate, and norsufentanil. Binding modes between the compounds and mu and delta-opioid receptors were studied by molecular docking method. The relationship between the molecular structural features and the analgesic activity was investigated by a quantitative structure-activity relationship model. The results of the molecular modeling studies and the in vivo analgesic activity suggested that the majority of the synthesized compounds were more potent than sufentanil and norsufentanil.
Subject(s)
Analgesics/chemical synthesis , Molecular Docking Simulation , Quantitative Structure-Activity Relationship , Sufentanil/analogs & derivatives , Acute Pain/drug therapy , Analgesics/metabolism , Analgesics/therapeutic use , Animals , Binding Sites , Male , Mice , Naloxone/chemistry , Naloxone/metabolism , Protein Structure, Tertiary , Receptors, Opioid, delta/chemistry , Receptors, Opioid, delta/metabolism , Receptors, Opioid, mu/chemistry , Receptors, Opioid, mu/metabolism , Sufentanil/chemistry , Sufentanil/metabolism , Sufentanil/therapeutic useABSTRACT
A rapid and sensitive method for the simultaneous determination of alfentanyl, sufentanyl and fentanyl (and its major metabolite norfentanyl) in urine was developed and validated. The method involved a liquid-liquid extraction in alkaline conditions, derivatization with pentafluoropropionic anhydride to improve the sensitivity for norfentanyl and subsequent analysis in GC/MS. The LODs are 0.08 ng ml(-1) for all substances (0.04 ng ml(-1) for alfentanyl). Intra- and inter-day precision coefficient of variation was always below 15%; mean relative error (accuracy) was always below 15%. The method was linear for all analytes, with quadratic regression of calibration curves always higher than 0.99. The method was applied to real samples of subjects who had received therapeutic doses of fentanyl, showing its suitability for the determination of low levels of these substances. The method was also applied to a subject whose death was attributed to fentanyl overdose.
Subject(s)
Alfentanil/analogs & derivatives , Alfentanil/urine , Fentanyl/analogs & derivatives , Gas Chromatography-Mass Spectrometry/methods , Sufentanil/analogs & derivatives , Sufentanil/urine , Alfentanil/chemistry , Alfentanil/toxicity , Calibration , Fentanyl/chemistry , Fentanyl/toxicity , Fentanyl/urine , Fluorocarbons/metabolism , Humans , Limit of Detection , Linear Models , Liquid-Liquid Extraction , Reproducibility of Results , Sufentanil/chemistry , Sufentanil/toxicityABSTRACT
An improved process for the synthesis of sufentanil with an overall yield of 26% is described. The reactive and high yielding N-debenzylation of the piperidine intermediate 7 using a mixture of Pd/C and Pd(OH)(2) was applied to other drug intermediates affording free amines in short reaction times. The deuterium-labeled sufentanil and the metabolite desmethylsufentanil were synthesized applying the optimized process.
Subject(s)
Analgesics/chemical synthesis , Sufentanil/analogs & derivatives , Sufentanil/chemical synthesis , Analgesics/chemistry , Analgesics/metabolism , Molecular Structure , Palladium/chemistry , Sufentanil/chemistryABSTRACT
The synthesis of an (18)F-labeled sufentanil analogue with apparent high mu-opioid receptor selectivity is reported. Intravenous injection of N-[4-(methoxymethyl)-1-[2-(2-thienyl)ethyl]-4-piperidinyl]-N-phenyl-2-(+/-)-[(18)F]fluoropropan-amide in mice resulted in high brain uptake and a regional brain activity distribution corresponding to the mu-opioid receptor expression pattern. The developed ligand is a promising tracer for extended protocols in mu-opioid receptor mapping and quantitation with positron emission tomography.
Subject(s)
Fluorine Radioisotopes , Radiopharmaceuticals/metabolism , Receptors, Opioid, mu/metabolism , Sufentanil/metabolism , Animals , Brain/diagnostic imaging , Brain/metabolism , Mice , Positron-Emission Tomography/methods , Sufentanil/analogs & derivativesABSTRACT
A sensitive, specific urinary assay for fentanyl, sufentanil, and alfentanil based on their N-dealkylated metabolites is described. Norfentanyl, norsufentanil-noralfentanil, and 2H5-norfentanyl are synthesized and characterized by standard analytical techniques. Derivatization of these secondary amines to yield the pentafluorobenzamides produces stable products with good gas chromatographic properties and unique, high-mass fragments in their mass spectra. These properties are utilized to develop a drug screening procedure based on gas chromatography-mass spectrometry to detect these major metabolites in human urine. The metabolites are isolated from urine samples by a liquid-liquid extraction procedure. The method allows for detection of metabolite concentrations as low as 0.3 ng/mL.
Subject(s)
Alfentanil/urine , Analgesics, Opioid/urine , Drug Residues/analysis , Fentanyl/urine , Gas Chromatography-Mass Spectrometry/methods , Narcotics/urine , Sufentanil/urine , Alfentanil/chemistry , Alfentanil/metabolism , Analgesics, Opioid/chemistry , Analgesics, Opioid/metabolism , Fentanyl/analogs & derivatives , Fentanyl/chemistry , Fentanyl/metabolism , Humans , Narcotics/chemistry , Narcotics/metabolism , Sensitivity and Specificity , Sufentanil/analogs & derivatives , Sufentanil/chemistry , Sufentanil/metabolismABSTRACT
Alfentanil, sufentanil, and fentanyl are synthetic opioids that are metabolized by oxidative N-dealkylation in the liver. We have previously shown that cytochrome P-450 3A4 (CYP3A4) contributes significantly to human liver microsomal alfentanil oxidation. Since identification of specific drug-metabolizing enzymes allows prediction of the variables affecting drug metabolism, the purpose of the present study was to identify the P-450 enzymes responsible for sufentanil and fentanyl metabolism in human liver microsomes. Microsomal preparations fortified with a reduced nicotinamide-adenine dinucleotide phosphate-generating system were incubated with 0.25 microM 3H-fentanyl or 3H-sufentanil. Rates of N-dealkylated metabolite formation significantly correlated with nifedipine oxidation activity (a marker of CYP3A4 activity) for fentanyl and sufentanil (r = 0.93 and 0.87, n = 18, respectively), but not with the oxidation activity for ethoxyresorufin (CYP1A2), S-mephenytoin (CYP2C19), bufuralol (CYP2D6), or chlorzoxazone (CYP2E1). Gestodene and troleandomycin (chemical inhibitors of CYP3A4) and antibody to CYP3A4 inhibited N-dealkylation of fentanyl and sufentanil. Chemical inhibitors of CYP2C, 2E1, and 2D6 did not inhibit N-dealkylation of fentanyl and sufentanil. Recombinant CYP3A4 expressed in Escherichia coli showed N-dealkylation activity of fentanyl and sufentanil, while expressed CYP1A2, 2C10, and 2E1 enzymes did not. We conclude that CYP3A4 is responsible for fentanyl and sufentanil N-dealkylation in vitro.