ABSTRACT
Magnetic surface imprinted polymer microspheres (Fe3O4@MIPs) were successfully synthesized via Pickering emulsion polymerization, utilizing N-Methylphenethylamine as a surrogate template for amphetamine-type drugs. Fe3O4@MIPs not only possessed excellent dispersibility and enough magnetic properties in aqueous solutions, but also displayed good selectivity towards six amphetamines, with an imprinting factor ranging from 1.8 to 2.6. The adsorption kinetics closely aligned with the pseudo-second-order model, and the adsorption efficiency exceeds 80 % for each amphetamine at equilibrium. Fe3O4@MIPs were then employed as the efficient adsorbents for the extraction of amphetamine drugs. Extraction parameters, including sample pH, the mass of adsorbent, and the type and volume of eluting solvent, were carefully optimized. In combination with the high performance liquid chromatography tandem triple quadrupole mass spectrometry (HPLC-MS/MS), a selective magnetic solid-phase extraction (MISPE) method utilizing Fe3O4@MIPs was developed for the detection of six amphetamines in water samples. The limits of detection and limits of quantitation were determined to be 5.2â¼23 ng L-1 and 17â¼77 ng L-1, respectively. Recoveries for the six target drugs from lake water and sewage samples fell within the range of 87.2â¼110 %. Additionally, the MISPE-HPLC-MS/MS method exhibited excellent repeatability, with a precision below 8.5 % at two spiking levels. The prepared Fe3O4@MIPs possessed the advantages of high selectivity, straightforward preparation, facile separation and good reusability, and was highly suitable for the efficient extraction of amphetamine-type substances in complex environmental water.
Subject(s)
Amphetamines , Limit of Detection , Microspheres , Solid Phase Extraction , Tandem Mass Spectrometry , Water Pollutants, Chemical , Chromatography, High Pressure Liquid/methods , Tandem Mass Spectrometry/methods , Water Pollutants, Chemical/analysis , Water Pollutants, Chemical/chemistry , Solid Phase Extraction/methods , Adsorption , Amphetamines/analysis , Amphetamines/isolation & purification , Amphetamines/chemistry , Polymers/chemistry , Molecular Imprinting/methods , Amphetamine/analysis , Amphetamine/chemistry , Molecularly Imprinted Polymers/chemistry , Liquid Chromatography-Mass SpectrometryABSTRACT
Among different substance classes, New Psychoactive Substances (NPS) comprise chiral amphetamines for stimulant and empathic effects. There is little knowledge in terms of clinical studies about possibly different effects of the two enantiomers of novel amphetamine derivatives. For this reason, there is a big demand for enantioseparation method development of this new substance class. Regarding gas chromatography, cyclodextrins proved to be effective for enantioseparation of NPS. In our attempt, an Astec® Chiraldex™ G-PN column containing 2,6-di-O-pentyl-3-propionyl-γ-cyclodextrin and a Lipodex™ D column containing heptakis-(2,6-di-O-pentyl-O-acetyl)-ß-cyclodextrin as chiral selector served as stationary phases in a Shimadzu GCMS-QP2010 SE system. Because of the special coating, maximum temperature is limited to 200 °C isothermal or 220 °C in programmed mode. To ensure detection, trifluoroacetic anhydride (TFAA) was used to increase sample volatility.1 As a result, 35 amphetamines were tested as their TFAA-derivatives. A screening method with a temperature gradient from 140 °C to 200 °C at a heating ramp of 1 °C per minute and final time of 5 min, showed baseline separation for seven and partial separations for 16 trifluoro acetylated amphetamines using the Chiraldex™ G-PN column. Six baseline and nine partial separations were observed with the Lipodex™ D column, respectively.
Subject(s)
Amphetamines , Stereoisomerism , Amphetamines/chemistry , Amphetamines/isolation & purification , Chromatography, Gas/methods , Cyclodextrins/chemistry , Temperature , Gas Chromatography-Mass Spectrometry/methodsABSTRACT
The use of controlled precursors for reaction optimisation is not always practical. One approach to limiting the use of controlled substances is to instead use 'model compounds'. Herein, two model compounds resembling norephedrine and ephedrine were selected based on their (i) structural similarity (i.e., presence of key functional groups) and (ii) availability from multiple suppliers without restriction. Model compounds 2-amino-1-phenylethanol and 2-(methylamino)-1-phenylethanol (halostachine), were compared to norephedrine and pseudoephedrine by firstly subjecting them to transformations known in the synthesis of amphetamines, and secondly, comparing the compounds using colourimetric spot tests, FTIR and NMR.
Subject(s)
Amphetamines , Central Nervous System Stimulants , Magnetic Resonance Spectroscopy , Spectroscopy, Fourier Transform Infrared , Amphetamines/chemistry , Central Nervous System Stimulants/chemistry , Humans , Ephedrine/chemistry , Colorimetry , Phenylpropanolamine/chemistry , Pseudoephedrine/chemistry , Models, ChemicalABSTRACT
Herein, the capture and separation properties of the deep eutectic solvent-functionalized magnetic graphene oxide/ZIF-67 composite (ZMG-DES) towards amphetamine-type drugs (MDMA, MAM and AM) from water were investigated. Kinetic and isotherm models showed that the adsorption behaviors were monolayer chemisorption. Batch experiment results showed that the maximal adsorption of MDMA (933.652 µgâ g-1) was 2.3 and 2.8 times higher than that of MAM (412.849 µgâ g-1) and AM (328.652 µgâ g-1), respectively, and this superiority remained consistent under varied environmental influences (pH, background ion and humic acid). Theoretical calculations and characterization analyses demonstrated the methylenedioxy group of MDMA led to the highly selective adsorption. Electrostatic potential (ESP) distribution indicated that the methylenedioxy added electron-rich areas and provided more adsorption sites. The Independent Gradient Model (IGMH) quantified the adsorption contribution of the functional groups in each system, which the contribution of the methylenedioxy reached 25.23%, significantly exceeding that of -NH- (18.80%) and benzene ring (20.76%), and proved that the H-bonds formed methylenedioxy enhanced adsorption. Furthermore, the Hirshfeld surface analysis proved that the methylenedioxy and -NH- of MDMA acted as H-bond acceptor and donor, respectively, which synergistically promoted the adsorption. The present study will help us to understand the structure-property relationship between amphetamine-type drugs and ZMG-DES.
Subject(s)
Amphetamines , Graphite , Imidazoles , Water Pollutants, Chemical , Zeolites , Adsorption , Amphetamines/chemistry , Deep Eutectic Solvents/chemistry , Graphite/chemistry , Kinetics , Metal-Organic Frameworks/chemistry , Models, Chemical , Water Pollutants, Chemical/chemistry , Water Pollutants, Chemical/analysisABSTRACT
The vaporization enthalpy, and vapor pressure as a function of temperature of N-ethylamphetamine, a substance used in the 1950s as an appetite suppressant and more currently abused as a designer drug, is reported. Its physical properties are compared to those of S (+)-N-methamphetamine, a substance whose physiological properties it mimics. A vaporization enthalpy of (62.4 ± 4.4) kJ·mol-1 and vapor pressure of (19 ± 11) Pa at T = 298.15 K has been evaluated by correlation gas chromatography. Results are compared to estimated values and to the limited amount of experimental property data available.
Subject(s)
Amphetamines/chemistry , Models, Chemical , Thermodynamics , Vapor Pressure , Chromatography, Gas , VolatilizationABSTRACT
Amphetamine derivatives have been used in a wide variety of pathologies because of their pharmacological properties as psychostimulants, entactogens, anorectics, and antidepressants. However, adverse cardiovascular effects (sympathomimetics) and substance abuse problems (psychotropic and hallucinogenic effects) have limited their use. 4-Methylthioamphetamine (MTA) is an amphetamine derivative that has shown to inhibit monoamine uptake and monoamine oxidase. However, the pharmacological characterization (neurochemical, behavioral, and safety) of its derivatives 4-ethylthioamphetamine (ETA) and 4-methylthio-phenil-2-butanamine (MT-But) have not been studied. In the current experiments, we show that ETA and MT-But do not increase locomotor activity and conditioned place preference with respect to MTA. At the neurochemical level, ETA and MT-But do not increase in vivo DA release in striatum, but ETA and MT-But affect the nucleus accumbens bioaccumulation of DA and DOPAC. Regarding cardiovascular effects, the administration of MTA and ETA increased the mean arterial pressure and only ETA significantly increases the heart rate. Our results show that the pharmacological and safety profiles of MTA are modulated by changing the methyl-thio group or the methyl group of the aminoethyl chain.
Subject(s)
3,4-Dihydroxyphenylacetic Acid/pharmacology , Amphetamine/pharmacology , Amphetamines/pharmacology , Corpus Striatum/drug effects , Dopamine/pharmacology , Nucleus Accumbens/drug effects , Amphetamines/chemistry , Animals , Behavior, Animal , Body Temperature , Ligands , Locomotion/drug effects , Male , Molecular Docking Simulation , Oxygen/chemistry , Rats , Rats, Sprague-Dawley , Serotonin Plasma Membrane Transport Proteins/chemistryABSTRACT
Amphetamine-based (Amph) drugs are metabolized in humans to their hydroxylamine (AmphNHOH) and nitroso (AmphNO) derivatives. The latter metabolites are known to bind to the Fe centers of cytochrome P450 and other heme enzymes to inhibit their activities. Although these AmphNHOH/AmphNO metabolites are present in vivo, their interactions with the blood protein hemoglobin (Hb) and the muscle protein (Mb) have been largely discounted due to a perception that the relatively small heme active sites of Hb and Mb will not be able to accommodate the large AmphNO group. We report the 2.15 Å resolution X-ray crystal structure of the AmphNO adduct of adult human hemoglobin as the Hb [α-FeIII(H2O)][ß-FeII(AmphNO)] derivative. We show that the binding of AmphNO to the ß subunit is enabled by an E helix movement and stabilization of ligand binding by H-bonding with the distal His63 residue. We also observe an AmphNHOH group in the Xe2 pocket in close proximity to the α heme site in this derivative. Additionally, UV-vis spectroscopy was used to characterize this and related wt and mutant Mb adducts. Importantly, our X-ray crystal structure of this Hb-nitrosoamphetamine complex represents the first crystal structure of a wild-type heme protein adduct of any amphetamine metabolite. Our results provide a framework for further studies of AmphNHOH/AmphNO interactions with Hb and Mb as viable processes that potentially contribute to the overall biological inorganic chemistry of amphetamine drugs.
Subject(s)
Amphetamines/metabolism , Hemoglobins/metabolism , Nitroso Compounds/metabolism , Amphetamines/chemistry , Binding Sites , Catalytic Domain , Crystallography, X-Ray , Hemoglobins/chemistry , Humans , Ligands , Molecular Structure , Nitroso Compounds/chemistry , Spectrophotometry, UltravioletABSTRACT
Interpretation of amphetamine-type stimulant (ATS) findings in urine samples can be challenging without chiral information. We present a sensitive enantioselective high-performance liquid chromatography-tandem mass spectrometry method for the quantification of (R)-amphetamine, (S)-amphetamine, (R)-methamphetamine, (S)-methamphetamine, (1R,2R)-pseudoephedrine, (1S,2S)-pseudoephedrine, (1R,2S)-ephedrine, (1S,2R)-ephedrine, (1R,2S)-norephedrine, (1S,2R)-norephedrine, (R)-cathinone, (S)-cathinone, and (1S,2S)-norpseudoephedrine (cathine) in urine. The method was successfully applied to more than 100 authentic urine samples from forensic casework. In addition, samples from a controlled self-administration of (1S,2S)-pseudoephedrine (Rinoral, 1200 mg within 6 days) were analyzed. The results strengthen the hypothesis that (1R,2S)-norephedrine is a minor metabolite of amphetamine and methamphetamine. We suggest cathine and (1S,2R)-norephedrine as minor metabolites of amphetamine racemate in humans. Small methamphetamine concentrations detected in samples with high concentrations of amphetamine could result from a metabolic formation by methylation of amphetamine although in samples with an (R)/(S) ratio for methamphetamine < 1 an additional (previous) (S)-methamphetamine consumption seems likely. Our data suggest that even amphetamine concentrations exceeding methamphetamine concentrations in urine can be caused by the biotransformation of methamphetamine to amphetamine as long as no (R)-amphetamine is detected. However, without chiral information, such findings might be (falsely) assumed as a co-consumption of both substances. Cathinone enantiomers detected in urine samples with high amphetamine concentrations can be interpreted as metabolites of amphetamine. In addition, the results of the self-administration study revealed that both cathinone enantiomers are minor metabolites of (1S,2S)-pseudoephedrine, which is the active ingredient of various medicines used for cold. The enantioselective analysis is a powerful tool to avoid the misinterpretation of ATS findings in urine samples.
Subject(s)
Alkaloids/analysis , Amphetamines/analysis , Chromatography, High Pressure Liquid/methods , Ephedrine/analysis , Alkaloids/chemistry , Alkaloids/metabolism , Amphetamines/chemistry , Amphetamines/metabolism , Central Nervous System Stimulants/analysis , Central Nervous System Stimulants/chemistry , Ephedrine/analogs & derivatives , Ephedrine/chemistry , Humans , Male , Middle Aged , Stereoisomerism , Tandem Mass Spectrometry/methodsABSTRACT
A simple and rapid pipette-tip solid-phase extraction (PT-SPE) procedure with derivatization prior to gas chromatography triple quadrupole mass spectrometry analysis is developed for the simultaneous determination of amphetamine (AMP), methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA) and 3,4-methylenedioxymethamphetamine (MDMA) in urine samples. The PT-SPE procedure using self-made sorbent could extract drugs within 6 min from 100-µL urine samples, requiring low solvent-consumption (<2.0 mL). Besides, the self-made pipette tip could be reused at least five times. Under optimized conditions, the recoveries of four amphetamines at spiked levels (low, medium and high) ranged from 87.7 to 110.4%, with relative standard deviations < 9.5%. The limit of detections and limit of quantifications for AMP, MA, MDA and MDMA were in the range of 2.52-8.25 ngâ mL-1 and 8.4-27.5 ngâ mL-1, respectively. Validation results show that the proposed method is suitable for the quantitation of amphetamines and has been successfully applied in the urine samples of suspected drug abusers.
Subject(s)
Amphetamines/urine , Gas Chromatography-Mass Spectrometry/methods , Solid Phase Extraction/methods , Tandem Mass Spectrometry/methods , Amphetamines/chemistry , Amphetamines/isolation & purification , Equipment Design , Humans , Limit of Detection , Linear Models , Reproducibility of Results , Solid Phase Extraction/instrumentationABSTRACT
Increasing reports of neurological and psychiatric complications due to psychostimulant synthetic cathinones (SCs) have recently raised public concern. However, the precise mechanism of SC toxicity is unclear. This paucity of understanding highlights the need to investigate the in-vitro toxicity and mechanistic pathways of three SCs: butylone, pentylone, and 3,4-Methylenedioxypyrovalerone (MDPV). Human neuronal cells of SH-SY5Y were cultured in supplemented DMEM/F12 media and differentiated to a neuronal phenotype using retinoic acid (10 µM) and 12-O-tetradecanoylphorbol-13-acetate (81 nM). Trypan blue and lactate dehydrogenase assays were utilized to assess the neurotoxicity potential and potency of these three SCs. To investigate the underlying neurotoxicity mechanisms, measurements included markers of oxidative stress, mitochondrial bioenergetics, and intracellular calcium (Ca2+), and cell death pathways were evaluated at two doses (EC15 and EC40), for each drug tested. Following 24 h of treatment, all three SCs exhibited a dose-dependent neurotoxicity, characterized by a significant (p < 0.0001 vs. control) production of reactive oxygen species, decreased mitochondrial bioenergetics, and increased intracellular Ca2+ concentrations. The activation of caspases 3 and 7 implicated the orchestration of mitochondrial-mediated neurotoxicity mechanisms for these SCs. Identifying novel therapeutic agents to enhance an altered mitochondrial function may help in the treatment of acute-neurological complications arising from the illicit use of these SCs.
Subject(s)
Alkaloids/pharmacology , Dopaminergic Neurons/cytology , Mitochondria/metabolism , Adenosine Triphosphate/metabolism , Alkaloids/chemistry , Amphetamines/chemistry , Amphetamines/pharmacology , Benzodioxoles/chemistry , Benzodioxoles/pharmacology , Calcium/metabolism , Caspase 3/metabolism , Caspase 7/metabolism , Cell Death/drug effects , Cell Line, Tumor , Dopaminergic Neurons/drug effects , Dopaminergic Neurons/metabolism , Energy Metabolism , Homeostasis/drug effects , Humans , Mitochondria/drug effects , Neurotoxins/toxicity , Oxidative Stress/drug effects , Oxygen Consumption/drug effects , Pyrrolidines/chemistry , Pyrrolidines/pharmacology , Reactive Oxygen Species/metabolism , Synthetic CathinoneABSTRACT
Here, we present a fully validated method using a hollow-fibre liquid-phase microextraction technique for the determination by gas chromatography-mass spectrometry (GC-MS) of amphetamine (AMP), methamphetamine (MET), fenproporex (FEN), 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyamphetamine (MDA) and 3,4-methylenedioxyethylamphetamine (MDEA) in whole blood. The validation parameters presented successful values within those recommended by the Scientific Working Group for Forensic Toxicology (SWGTox) in the Standard Practices for Method Validation in Forensic Toxicology. The limits of detection ranged from 1 to 3 ng/mL, and the limits of quantification ranged from 2 to 5 ng/mL. The determination coefficients (r2) ranged from 0.990 to 0.997, and the method presented good intraday and interday accuracy (from 90.4% to 97.2%) and satisfactory recovery (from 68% to 110%). No carryover was observed. The heteroscedasticity was tested, and only AMP presented homoscedasticity. Weighting factors were applied to correct the linearity of MET (1/x2), MDA (1/x), FEN (1/x1/2), MDMA (1/x2) and MDEA (1/y). Dilution integrity was tested at ratios of 1:2, 1:5 and 1:10, and all maintained intraday precision (from 94.9% to 99.3%) and interday precision (from 89.4% to 94.9%). The validated method was applied to six real whole blood samples from individuals suspected of consuming ecstasy, and MDMA, MDA and amphetamine were successfully identified and quantified.
Subject(s)
Amphetamines/blood , Gas Chromatography-Mass Spectrometry/methods , Liquid Phase Microextraction/methods , Amphetamines/chemistry , Amphetamines/isolation & purification , Forensic Toxicology , Green Chemistry Technology , Humans , Limit of Detection , Linear Models , Reproducibility of ResultsABSTRACT
3-Fluoroamphetamine (also called PAL-353) is a synthetic amphetamine analog that has been investigated for cocaine use disorder (CUD), yet no studies have characterized its pharmacokinetics (PK). In the present study, we determined the PK of PAL-353 in male Sprague Dawley rats following intravenous bolus injection (5 mg/kg). Plasma samples were analyzed using a novel bioanalytical method that coupled liquid-liquid extraction and LC-MS/MS. The primary PK parameters determined by WinNonlin were a C0 (ng/mL) of 1412.09 ± 196.12 and a plasma half-life of 2.27 ± 0.67 h. As transdermal delivery may be an optimal approach to delivering PAL-353 for CUD, we assessed its PK profile following application of 50 mg of transdermal gel (10% w/w drug over 5 cm2). The 10% w/w gel resulted in a short lag time, sustained delivery, and a rapid clearance in plasma immediately after removal. The rodent PK data were verified by examining in vitro permeation through human epidermis mounted on Franz diffusion cells. An in vitro-in vivo correlation (IVIVC) analysis was performed using the Phoenix IVIVC toolkit to assess the predictive relationship between rodent and human skin absorption/permeation. The in vitro permeation study revealed a dose-proportional cumulative and steady-state flux with ~ 70% of drug permeated. The fraction absorbed in vivo and fraction permeated in vitro showed a linear relationship. In conclusion, we have characterized the PK profile of PAL-353, demonstrated that it has favorable PK properties for transdermal administration for CUD, and provided preliminary evidence of the capacity of rodent data to predict human skin flux.
Subject(s)
Amphetamines/administration & dosage , Amphetamines/pharmacokinetics , Epidermis/chemistry , Plasma/chemistry , Administration, Cutaneous , Administration, Intravenous , Amphetamines/chemistry , Animals , Chromatography, Liquid , Half-Life , Humans , Male , Rats , Rats, Sprague-Dawley , Skin Absorption , Tandem Mass SpectrometryABSTRACT
The pre-precursor market and the clandestine production of amphetamine-type stimulants (ATS) has become more diverse in recent years. Besides α-phenylacetoacetonitrile (APAAN) and α-phenylacetoacetamide (APAA), glycidic acid derivatives and methyl α-phenylacetoacetate (MAPA) are gaining importance. This conclusion is based on seizure data of police and customs. However, analytical data are needed to confirm and quantify the actual prevalence of new pre-precursors by elucidating the percentage of seized ATS that have been produced from them. A recent study showed that APAAN use is currently declining, which supports the view that new pre-precursors are being used. In this study, several conversion procedures using different batches of glycidic acid derivatives and a complete Leuckart reaction to produce amphetamine were carried out. The resulting organic phases were analyzed using gas chromatography - mass spectrometry to identify possible marker compounds. Three marker compounds were discovered and characterized using mass spectra and nuclear magnetic resonance spectroscopy. They were identified as phenyl-1-propanone, N-(1-phenylpropyl)formamide and 1-phenylpropan-1-amine. Their prevalence was investigated by searching the markers in an amphetamine impurity profiling database to determine to what extent they occurred in amphetamine samples from recent years. Data from the central German amphetamine profiling database of more than 250 cases were used for this purpose. The yearly occurrence of the three glycidate marker compounds was determined going back as far as 2009, revealing an increasing trend from 2016 on. This article presents experimental proof that APAAN is currently being replaced by other pre-precursors, such as glycidic acid derivatives.
Subject(s)
Amphetamines/chemistry , Central Nervous System Stimulants/chemistry , Epoxy Compounds/chemistry , Propionates/chemistry , Amphetamines/chemical synthesis , Central Nervous System Stimulants/chemical synthesis , Chemistry Techniques, Synthetic , Databases, Pharmaceutical , Drug Contamination , Epoxy Compounds/chemical synthesis , Gas Chromatography-Mass Spectrometry , Magnetic Resonance Spectroscopy , Propionates/chemical synthesisABSTRACT
The present paper describes an original method for the online preconcentration and analysis of ring-substituted amphetamines in urine samples, used on the integration of robot-assisted hollow fiber liquid-phase microextraction (HF-LPME), high-performance liquid chromatography (HPLC), and fluorescence detection (FLD). A lab-made autosampler, actuating a 100-µL syringe and equipped with a three-way solenoid microvalve, allowed the acceptor phase to flow through and be withdrawn from the lumen fiber, enabling the automated online transference of the enriched acceptor phase for chromatographic analysis, through a six-port switching valve. The developed online HF-LPME-LC/FLD method demonstrated high analytical throughput and confidence, facilitating the efficient extraction and determination of the target analytes, with minimal solvent consumption and sample manipulation, in a straightforward way. Sample cleanup, analyte uptake, and analysis were carried out in 14.5 min. Under optimal conditions, automated online HF-LPME showed excellent linearity, precision, and trueness, obtaining intraday RSDs between 2.9 and 9.2% (n = 6) and interday RSDs between 5.3 and 9.3% (n = 6). Enrichment factors (EFs) ranged between 14.2 and 15.7, extraction recoveries (ERs) ranged between 17.7 and 19.5%, and the limits of detection (S/N = 3) were 2.0, 3.0, and 3.0 µg L-1 for MDA, MDMA, and MDEA, respectively. The method proved to be an effortless, rapid, reliable, and environment-friendly approach for the determination of drug abuse in urine samples. Graphical abstract.
Subject(s)
Amphetamines/urine , Chromatography, High Pressure Liquid/methods , Liquid Phase Microextraction/methods , Amphetamines/chemistry , Humans , Limit of Detection , Reproducibility of Results , Spectrometry, Fluorescence/methodsABSTRACT
Here, we demonstrate a supramolecular latching tool for bio-orthogonal noncovalent anchoring of small synthetic molecules in live animal models using a fully synthetic high-affinity binding pair between cucurbit[7]uril (CB[7]) and adamantylammonium (AdA). This supramolecular latching system is small (â¼1 kDa), ensuring efficient uptake into cells, tissues, and whole organisms. It is also chemically robust and resistant to enzymatic degradation and analogous to well-characterized biological systems in terms of noncovalent binding. Occurrence of fluorescence resonance energy transfer (FRET) between cyanine 3-CB[7] (Cy3-CB[7]) and boron-dipyrromethene 630/650X-AdA (BDP630/650-AdA) inside a live worm (Caenorhabditis elegans) indicates efficient in situ high-affinity association between AdA and CB[7] inside live animals. In addition, selective visualization of a cancer site of a live mouse upon supramolecular latching of cyanine 5-AdA (Cy5-AdA) on prelocalized CB[7]-conjugating antibody on the cancer site demonstrates the potential of this synthetic system for in vivo cancer imaging. These findings provide a fresh insight into the development of new chemical biology tools and medical therapeutic systems.
Subject(s)
Diagnostic Imaging/instrumentation , Fluorescence Resonance Energy Transfer/instrumentation , Neoplasms/diagnostic imaging , Adamantane/analogs & derivatives , Adamantane/chemistry , Amphetamines/chemistry , Animals , Caenorhabditis elegans , Cell Line, Tumor , Female , Fluorescent Dyes/chemistry , Humans , Mice , Mice, Inbred BALB CABSTRACT
Synthetic cathinones also known as ß-keto amphetamines are a new group of recreational designer drugs. We aimed to evaluate the cytotoxic potential of thirteen cathinones lacking the methylenedioxy ring and establish a putative structure-toxicity profile using differentiated SH-SY5Y cells, as well as to compare their toxicity to that of amphetamine (AMPH) and methamphetamine (METH). Cytotoxicity assays [mitochondrial 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) reduction and lysosomal neutral red (NR) uptake] performed after a 24-h or a 48-h exposure revealed for all tested drugs a concentration-dependent toxicity. The rank order regarding the concentration that promoted 50 % of toxicity, at 24â¯h exposure, by the MTT assay was: 3,4-dimethylmethcathinone (3,4-DMMC) > METHâ¯>â¯mephedrone ≈ α-pyrrolidinopentiophenoneâ¯>â¯AMPH ≈ methedroneâ¯>â¯pentedroneâ¯>â¯buphedrone ≈ flephedrone >α-pyrrolidinobutiophenoneâ¯>â¯methcathinone ≈ N-ethylcathinone >α-pyrrolidinopropiophenone >N,N-dimethylcathinone ≈ amfepramone. Apoptotic cell death signs were seen for all studied cathinones. 3,4-DMMC, methcathinone and pentedrone triggered autophagy activation, as well as increased reactive oxygen species production, and N-acetyl-L-cysteine (NAC) totally prevented that rise. Importantly, NAC was also able to prevent the cytotoxicity promoted by 6 tested drugs, ruling for an involvement of oxidative stress in the toxic events observed. The increased lipophilic chain on the alpha carbon, the presence and the high steric volume occupied by the substituents on the aromatic ring, and the substitution of the pyrrolidine ring by its secondary amine analogue have proved to be key points for the cytotoxicity profile of these cathinones. The structure-toxicity relationship established herein may enlighten future human relevant mechanistic studies, and future clinical approaches on intoxications.
Subject(s)
Alkaloids/toxicity , Amphetamines/toxicity , Neurons/drug effects , Alkaloids/chemistry , Amphetamines/chemistry , Apoptosis/drug effects , Autophagy/drug effects , Cell Line, Tumor , Dose-Response Relationship, Drug , Humans , Microscopy, Phase-Contrast , Neurons/ultrastructure , Propiophenones/toxicity , Reactive Oxygen Species/metabolism , Structure-Activity RelationshipABSTRACT
Much evidence demonstrates the antinociceptive effect of magnetic fields (MFs). However, the analgesic action mechanism of the electromagnetic field (EMF) is not exactly understood. The aim of the present study was to investigate the effects of 5-HT1 and 5-HT2 receptor agonists (serotonin HCl and 2,5-dimethoxy-4-iodoamphetamine [DOI] hydrochloride) on EMF-induced analgesia. In total, 66 adult male Wistar albino rats with an average body mass of 225 ± 13 g were used in this study. The animals were subjected to repeated exposures of alternating 50 Hz and 5 mT EMF for 2 h a day for 15 days. Prior to analgesia tests, serotonin HCl (5-HT1 agonist) 4 mg/kg, WAY 100635 (5-HT1 antagonist) 0.04 mg/kg, DOI hydrochloride (5-HT2 receptor agonist) 4 mg/kg, and SB 204741 (5-HT2 antagonist) 0.5 mg/kg doses were injected into rats. For statistical analysis of the data, analysis of variance was used and multiple comparisons were determined by Tukey's test. Administration of serotonin HCl MF (5 mT)-exposed rats produced a significant increase in percent maximal possible effect (% MPE) as compared with EMF group (P < 0.05). On the contrary, injection of WAY 100635 to MF-exposed rats produced a significant decrease in analgesic activity (P < 0.05). Similarly, the administration of DOI hydrochloride significantly increased % MPE values as compared with the EMF group while SB 204741 reduced it (P < 0.05). In conclusion, our results suggested that serotonin 5-HT1 and 5-HT2 receptors play an important role in EMF-induced analgesia; however, further research studies are necessary to understand the mechanism. Bioelectromagnetics. 2019;40:319-330. © 2019 Bioelectromagnetics Society.
Subject(s)
Amphetamines/pharmacology , Analgesia , Electromagnetic Fields , Serotonin Receptor Agonists/pharmacology , Amphetamines/chemistry , Animals , Male , Pain Management , Rats, Wistar , Serotonin/metabolism , Serotonin Receptor Agonists/chemistryABSTRACT
Captagon, known by its genetic name Fenethylline, is an addictive drug that complicates the War on Drugs. Captagon has a strong CNS stimulating effect than its primary metabolite, Amphetamine. However, multi-targets issues associated with the drug and metabolites as well as its underlying mechanisms have not been fully defined. In the present work, we applied our established drug-abuse chemogenomics-knowledgebase systems pharmacology approach to conduct targets/off-targets mapping (SP-Targets) investigation of Captagon and its metabolites for hallucination addiction, and also analyzed the cell signaling pathways for both Amphetamine and Theophylline with data mining of available literature. Of note, Amphetamine, an agonist for trace amine-associated receptor 1 (TAAR1) with enhancing dopamine signaling (increase of irritability, aggression, etc.), is the main cause of Captagon addiction; Theophylline, an antagonist that blocks adenosine receptors (e.g. A2aR) in the brain responsible for restlessness and painlessness, may attenuate the behavioral sensitization caused by Amphetamine. We uncovered that Theophylline's metabolism and elimination could be retarded due to competition and/or blockage of the CYP2D6 enzyme by Amphetamine; We also found that the synergies between these two metabolites cause Captagon's psychoactive effects to act faster and far more potently than those of Amphetamine alone. We carried out further molecular docking modeling and molecular dynamics simulation to explore the molecular interactions between Amphetamine and Theophylline and their important GPCRs targets, including TAAR1 and adenosine receptors. All of the systems pharmacology analyses and results will shed light insight into a better understanding of Captagon addiction and future drug abuse prevention.
Subject(s)
Amphetamines/chemistry , Cytochrome P-450 CYP2D6 Inhibitors/chemistry , Cytochrome P-450 CYP2D6/chemistry , Molecular Docking Simulation , Molecular Dynamics Simulation , Receptor, Adenosine A2A/chemistry , Receptors, G-Protein-Coupled/chemistry , Substance-Related Disorders , Theophylline/analogs & derivatives , Cytochrome P-450 CYP2D6/metabolism , Humans , Receptor, Adenosine A2A/metabolism , Receptors, G-Protein-Coupled/metabolism , Theophylline/chemistryABSTRACT
RATIONALE: Synthetic cathinones continue to emerge in recreational drug markets worldwide. 1-(1,3-Benzodioxol-5-yl)-2-(methylamino)butan-1-one (butylone) and 1-(1,3-benzodioxol-5-yl)-2-(methylamino)pentan-1-one (pentylone) are derivatives of the cathinone compound, 1-(1,3-benzodioxol-5-yl)-2-(methylamino)propan-1-one (methylone), that are being detected in drug products and human casework. OBJECTIVES: The purpose of the present study was to examine the neuropharmacology of butylone and pentylone using in vitro and in vivo methods. METHODS: In vitro uptake and release assays were carried out in rat brain synaptosomes and in cells expressing human dopamine transporters (DAT) and 5-HT transporters (SERT). In vivo microdialysis was performed in the nucleus accumbens of conscious rats to assess drug-induced changes in neurochemistry. RESULTS: Butylone and pentylone were efficacious uptake blockers at DAT and SERT, though pentylone was more DAT-selective. Both drugs acted as transporter substrates that evoked release of [3H]5-HT at SERT, while neither evoked release at DAT. Consistent with the release data, butylone and pentylone induced substrate-associated inward currents at SERT but not DAT. Administration of butylone or pentylone to rats (1 and 3 mg/kg, i.v.) increased extracellular monoamines and motor activity, but pentylone had weaker effects on 5-HT and stronger effects on motor stimulation. CONCLUSIONS: Our data demonstrate that increasing the α-carbon chain length of methylone creates "hybrid" transporter compounds which act as DAT blockers but SERT substrates. Nevertheless, butylone and pentylone elevate extracellular dopamine and stimulate motor activity, suggesting both drugs possess significant risk for abuse.
Subject(s)
Alkaloids/pharmacology , Amphetamines/pharmacology , Central Nervous System Stimulants/pharmacology , Dopamine Antagonists/pharmacology , Dopamine Plasma Membrane Transport Proteins/antagonists & inhibitors , Dopamine Plasma Membrane Transport Proteins/metabolism , Synthetic Drugs/pharmacology , 3,4-Methylenedioxyamphetamine/analogs & derivatives , 3,4-Methylenedioxyamphetamine/chemistry , 3,4-Methylenedioxyamphetamine/pharmacology , Alkaloids/chemistry , Amphetamines/chemistry , Animals , Central Nervous System Stimulants/chemistry , Dopamine Antagonists/chemistry , Dose-Response Relationship, Drug , HEK293 Cells , Humans , Male , Methamphetamine/analogs & derivatives , Methamphetamine/chemistry , Methamphetamine/pharmacology , Nucleus Accumbens/drug effects , Nucleus Accumbens/metabolism , Rats , Rats, Sprague-Dawley , Synaptosomes/drug effects , Synaptosomes/metabolism , Synthetic Drugs/chemistryABSTRACT
BACKGROUND: The appearance of stimulant-class new psychoactive substances (NPS) is a frequent and significant problem in our society. Cathinone variants are often sold illegally as 3,4-methylenedioxy methamphetamine ("ecstasy") or disguised for legal sale using misleading names such as "bath salts" and carry the risk of promoting disruptive mental states, addiction, and fatal overdose. The principal targets of these recreational drugs are monoamine transporters expressed in catecholaminergic and serotonergic neurons. Some transporter ligands can be transported into cells, where they can promote a massive release of neurotransmitters through reverse transport, and others can block uptake. A ligand's dopamine vs. serotonin transporter selectivity, potency, and activity as a substrate or blocker can help elucidate the abuse liability and subjective effects of a drug. OBJECTIVES: Here, we describe the discovery, development, and validation of an emerging methodology for compound activity assessment at monoamine transporters. KEY FINDINGS: Substrates generate inward electrical currents through transporters and can depolarize the plasma membrane, whereas blockers work as a "cork in a bottle" and function as antagonists. Voltage-gated Ca2+ channels were co-expressed with monoamine transporters in cultured cells and used to measure fluctuations of the membrane electrical potential. In this system, substrates of monoamine transporters produce reliable dose-dependent Ca2+ signals, while blockers hinder them. DISCUSSION: This system constitutes a novel use of voltage-gated Ca2+ channels as biosensors for the purpose of characterizing ligand activity at monoamine transporters using fluorimetry. This approach in combination with in vivo evaluations of drugs' abuse-related effects is a powerful strategy for anticipating potential stimulant-class NPS.