piperidines and 2-methyl-1-1-diphenyl-3-(1-piperidinyl)-1-propanol

Metabolism studies play an important role in clinical and forensic toxicology. Because of potential species differences in metabolism, human samples are best suitable for elucidating metabolism. However, in the case of new psychoactive substances (NPS), human samples of controlled studies are not available. Primary human hepatocytes have been described as gold standard for in vitro metabolism studies, but there are some disadvantages such as high costs, limited availability, and variability of metabolic enzymes. Therefore, the aim of our study was to investigate and compare the metabolism of six methylenedioxy derivatives (MDMA, MDBD, butylone, MDPPP, MDPV, MDPB) and two bioisosteric analogues (5-MAPB, 5-API) using pooled human liver microsomes (pHLM) combined with cytosol (pHLC) or pooled human liver S9 fraction (pS9) all after addition of co-substrates for six phase I and II reactions. In addition, HepaRG and HepG2 cell lines were used. Results of the different in vitro tools were compared to each other, to corresponding published data, and to metabolites identified in human urine after consumption of MDMA, MDPV, or 5-MAPB. Incubations with pHLM plus pHLC showed similar results as pS9. A more cost efficient model for prediction of targets for toxicological screening procedures in human urine should be identified. As expected, the incubations with HepaRG provided better results than those with HepG2 concerning number and signal abundance of the metabolites. Due to easy handling without special equipment, incubations with pooled liver preparations should be the most suitable alternative to find targets for toxicological screening procedures for methylenedioxy derivatives and bioisosteric analogues.

Topics: Cell Line; Humans; Liver; Microsomes, Liver; N-Methyl-3,4-methylenedioxyamphetamine; Piperidines

In vitro cytochrome P450 (CYP) inhibition assays are common approaches for testing the inhibition potential of drugs for predicting potential interactions. In contrast to marketed medicaments, drugs of abuse, particularly the so-called novel psychoactive substances, were not tested before distribution and consumption. Therefore, the inhibition potential of methylenedioxy-derived designer drugs (MDD) of different drug classes such as aminoindanes, amphetamines, benzofurans, cathinones, piperazines, pyrrolidinophenones, and tryptamines should be elucidated. The FDA-preferred test substrates, split in two cocktails, were incubated with pooled human liver microsomes and analysed after protein precipitation using LC-high-resolution-MS/MS. IC50 values were determined of MDD showing more than 50 % inhibition in the prescreening. Values were calculated by plotting the relative metabolite concentration formed over the logarithm of the inhibitor concentration. All MDD showed inhibition against CYP2D6 activity and most of them in the range of the clinically relevant CYP2D6 inhibitors quinidine and fluoxetine. In addition, the beta-keto compounds showed inhibition of the activity of CYP2B6, 5,6-MD-DALT of CYP1A2 and CYP3A, and MDAI of CYP2A6, all in the range of clinically relevant inhibitors. In summary, all MDD showed inhibition of the activity of CYP2D6, six of CYP1A2, three of CYP2A6, 13 of CYP2B6, two of CYP2C9, six of CYP2C19, one of CYP2E1, and six of CYP3A. These results showed that the CYP inhibition by MDD might be clinically relevant, but further studies are needed for final conclusions.

Topics: 3,4-Methylenedioxyamphetamine; Cytochrome P-450 CYP2D6 Inhibitors; Cytochrome P-450 Enzyme Inhibitors; Designer Drugs; Drug Interactions; Humans; Inhibitory Concentration 50; Microsomes, Liver; Piperidines; Toxicity Tests

This case report is considered exempt from University of California-San Diego Investigational Review Board.. There is a limited published experience detailing detection and toxicity of multiple novel psychoactive substances. We report a case of a patient with prolonged psychosis who had JWH-072, cannabicyclohexanol, 3',4'-methylenedioxy-α-pyrrolidinopropiophenone (MDPPP) and methylenedioxyamphetamine (MDA) identified in multiple biological samples.. An 18-year-old man presented to the emergency department (ED) with acute onset psychosis after allegedly smoking "spice." Due to agitation and psychosis refractory to multiple medications, a lumbar puncture was performed and he was admitted. All blood, urine, and CSF (cerebral spinal fluid) testing was normal. He remained psychotic for almost 1 week. MDPPP, JWH-072 and MDA were detected in initial blood, urine, and CSF samples. Cannabicyclohexanol was detected only in his serum.. JWH-072 is a cannabinoid-2 receptor (CB-2) agonist which has not been reported previously in the literature. Its clinical effects are unknown. Cannabicyclohexanol is a known component of "spice" products and has been associated with agitation and psychosis. MDPPP and MDA are designer phenylethylamines likely to cause agitation and sympathomimetic symptoms. Simultaneous detection of novel psychoactive substances in multiple biological fluids has not been previously reported. This case suggests that the interaction of these particular substances may be associated with prolonged psychosis.

Topics: Adolescent; Cannabinoids; Cyclohexanols; Designer Drugs; Drug Interactions; Drug Therapy, Combination; Humans; Male; N-Methyl-3,4-methylenedioxyamphetamine; Phenols; Piperidines; Psychoses, Substance-Induced; Substance Abuse Detection

The metabolism of 3',4'-methylenedioxy-a-pyrrolidinopropiophenone (MDPPP), a novel designer drug, to its demethylenated major metabolite 3',4'-dihydroxy-pyrrolidinopropiophenone (di-HO-PPP) was studied in pooled human liver microsomes (HLM) and in cDNA-expressed human hepatic cytochrome P450 (CYP) enzymes. CYP2C19 catalysed the demethylenation with apparent Km and Vmax values of 120.0+/-13.4 microM and 3.2+/-0.1 pmol/min/pmol CYP, respectively (mean+/-standard deviation). CYP2D6 catalysed the demethylenation with apparent Km and Vmax values of 13.5+/-1.5 microM and 1.3+/-0.1 pmol/min/pmol CYP, respectively. HLM exhibited a clear biphasic profile with an apparent Km,1 value of 7.6+/-9.0 and a Vmax,1 value of 11.1+/-3.6 pmol/min/mg protein, respectively. Percentages of intrinsic clearances of MDPPP by specific CYPs were calculated using the relative activity factor (RAF) approach with (S)-mephenytoin-4'-hydroxylation or bufuralol-1'-hydroxylation as index reactions for CYP2C19 or CYP2D6, respectively. MDPPP, di-HO-PPP and the standard 4'-methyl-pyrrolidinohexanophenone (MPHP) were separated and analysed by liquid chromatography-mass spectrometry in the selected-ion monitoring (SIM) mode. The CYP2D6-specific chemical inhibitor quinidine (3 microM) significantly (p<0.001) inhibited di-HO-PPP formation by 75.8%+/-1.7% (mean+/-standard error of the mean) in incubation mixtures with HLM and 2 microM MDPPP. It can be concluded from the data obtained from kinetic and inhibition studies that polymorphically expressed CYP2D6 and CYP2C19 are almost equally responsible for MDPPP demethylenation.

Topics: Cells, Cultured; Cytochrome P-450 Enzyme System; Enzyme Activation; Humans; Kinetics; Metabolic Clearance Rate; Microsomes, Liver; Piperidines

Racemic 2-methyl-1,1-diphenyl-3-(1-piperidyl)-1-propanol (4) and 2-methyl-1,1-diphenyl-3-(1-pyrrolidyl)-1-propanol (5) have been synthesized and optically resolved employing (R)- and (S)-mandelic acid, respectively, as resolving agents. The absolute configurations of the enantiomers of 4 and 5 have been established as (-)-(R) and (+)-(S) by crystal structure analyses of (-)-(R)-2-methyl-1,1-diphenyl-3-(1-piperidinio)-1-propanol (+)-(S)-mandelate and (+)-(S)-2-methyl-1,1-diphenyl-3-(1-pyrrolidinio)-1-propanol (-)-(R)-mandelate. The corresponding methiodides have been prepared.

Topics: Crystallography; Mandelic Acids; Molecular Conformation; Parasympatholytics; Piperidines; Pyrrolidines; Stereoisomerism