Simple and selective screening method for the synthetic cathinone MDPT in forensic samples using carbon nanofiber screen-printed electrodes.

3',4'-Methylenedioxy-N-tert-butylcathinone (MDPT), also known as tBuONE or D-Tertylone, is a synthetic cathinone (SC) frequently abused for recreational purposes due to its potent stimulant effects and similarity to illegal substances like methamphetamine and ecstasy. The structural diversity and rapid introduction of new SC analogs to the market poses significant challenges for law enforcement and analytical methods for preliminary screening of illicit drugs. In this work, we present, for the first time, the electrochemical detection of MDPT using screen-printed electrodes modified with carbon nanofibers (SPE-CNF). MDPT exhibited three electrochemical processes (two oxidations and one reduction) on SPE-CNF. The proposed method for MDPT detection was optimized in 0.2 mol L-1 Britton-Robinson buffer solution at pH 10.0 using differential pulse voltammetry (DPV). The SPE-CNF showed a high stability for electrochemical responses of all redox processes of MDPT using the same or different electrodes, with relative standard deviations less than 4.7% and 1.5% (N = 3) for peak currents and peak potentials, respectively. Moreover, the proposed method provided a wide linear range for MDPT determination (0.90-112 µmol L-1) with low LOD (0.26 µmol L-1). Interference studies for two common adulterants, caffeine and paracetamol, and ten other illicit drugs, including amphetamine-like compounds and different SCs, showed that the proposed sensor is highly selective for the preliminarily identification of MDPT in seized forensic samples. Therefore, SPE-CNF with DPV can be successfully applied as a fast and simple screening method for MDPT identification in forensic analysis, addressing the significant challenges posed by the structural diversity of SCs.

Allosteric Binding of MDMA to the Human Serotonin Transporter (hSERT) via Ensemble Binding Space Analysis with ΔG Calculations, Induced Fit Docking and Monte Carlo Simulations.

Despite the recent promising results of MDMA (3,4-methylenedioxy-methamphetamine) as a psychotherapeutic agent and its history of misuse, little is known about its molecular mode of action. MDMA enhances monoaminergic neurotransmission in the brain and its valuable psychoactive effects are associated to a dual action on the 5-HT transporter (SERT). This drug inhibits the reuptake of 5-HT (serotonin) and reverses its flow, acting as a substrate for the SERT, which possesses a central binding site (S1) for antidepressants as well as an allosteric (S2) one. Previously, we characterized the spatial binding requirements for MDMA at S1. Here, we propose a structure-based mechanistic model of MDMA occupation and translocation across both binding sites, applying ensemble binding space analyses, electrostatic complementarity, and Monte Carlo energy perturbation theory. Computed results were correlated with experimental data (r = 0.93 and 0.86 for S1 and S2, respectively). Simulations on all hSERT available structures with Gibbs free energy estimations (ΔG) revealed a favourable and pervasive dual binding mode for MDMA at S2, i.e., adopting either a 5-HT or an escitalopram-like orientation. Intermediate ligand conformations were identified within the allosteric site and between the two sites, outlining an internalization pathway for MDMA. Among the strongest and more frequent interactions were salt bridges with Glu494 and Asp328, a H-bond with Thr497, a π-π with Phe556, and a cation-π with Arg104. Similitudes and differences with the allosteric binding of 5-HT and antidepressants suggest that MDMA may have a distinctive chemotype. Thus, our models may provide a framework for future virtual screening studies and pharmaceutical design and to develop hSERT allosteric compounds with a unique psychoactive MDMA-like profile.

Isquemia miocárdica por consumo de dibutilona / Myocardial ischemia from dibutylone use / Isquemia miocárdica por uso de dibutilona

Resumen: Las catinonas sintéticas son drogas estimulantes de síntesis, comercializadas en el mercado ilegal por sus propiedades estimulantes y perturbadoras del sistema nervioso central, para su uso recreativo, solas, o como adulterantes de cocaína y derivados anfetamínicos. A nivel mundial existe un número creciente de reportes de intoxicaciones agudas y complicaciones por consumo de estas sustancias. En Uruguay fueron detectadas por primera vez en el año 2015, existiendo una información epidemiológica muy limitada. Las catinonas sintéticas no se detectan por las técnicas inmunocromatográficas disponibles en los laboratorios de los servicios de urgencias de nuestro país. Se describe el primer caso de intoxicación por una catinona sintética (dibutilona) en Uruguay, en un paciente de 45 años, por consumo de un polvo vendido como "éxtasis", presentando como complicación un síndrome coronario agudo, con una buena evolución posterior. La confirmación diagnóstica se realizó mediante su detección en orina por cromatografía de gases acoplada a espectrometría de masas. La estructura química de la dibutilona y su mecanismo de acción explican las manifestaciones clínicas y la complicación isquémica por vasoespasmo coronario. En ausencia de una noción clara de exposición y ante la situación epidemiológica actual, la presencia de un cuadro clínico compatible con una droga estimulante y resultado negativo para cocaína y anfetaminas en estudios inmunocromatográficos, debe hacer plantear la sospecha de otros estimulantes de síntesis como las catinonas, tal como ocurrió en este caso.

Summary: Synthetic cathinone are stimulating synthetic drugs used for recreational purposes on their own or as adulterants in cocaine and amphetamines derivatives that are traded in illegal markets given their stimulating and disturbing properties on the central nervous system. Globally, there is a growing number of acute intoxications and complications as a consequence of consumption of these substances. In Uruguay they were first detected in 2015, there being limited epidemiological data. Synthetic cathinone are not detected by the immunochromatographic tests available at the laboratories of the emergency services in our country. The study describes the first case of intoxication by a synthetic cathinone (dibutylone) in Uruugay, in a 45 year old patient as a result of powder sold as "ecstasy". The patient presented an acute coronary syndrome, subsequent evolution being good. Diagnostic confirmation was made in urine using gas chromatography coupled with mass spectrometry. The chemical structure of dibutylone and its action mechanism explain the clinical manifestations and the coronary vasospasm causing ischemia. When there is no clear sign of a patient having been exposed to this substance and upon the current epidemiological state, the presence of clinical symptoms that are compatible with a stimulating drug and a negative result for cocaine and amphetamines in immunochromatographic tests must lead us to be suspicious about the presence of synthetic stimulating substances as cathinone, as in the case studied.

Resumo: Catinonas sintéticas são drogas estimulantes sintéticas, comercializadas no mercado ilegal por suas propriedades estimulantes e perturbadoras do sistema nervoso central, para uso recreativo isoladamente ou como adulterantes da cocaína e derivados de anfetaminas. Em todo o mundo cresce o número de relatos de intoxicações agudas e complicações decorrentes do uso dessas substâncias. No Uruguai, foram detectados pela primeira vez em 2015 com informações epidemiológicas muito limitadas. As catinonas sintéticas não são detectadas pelas técnicas imunocromatográficas disponíveis nos laboratórios de pronto-socorro de nosso país. Descreve-se o primeiro caso de intoxicação por catinona sintética (dibutilona) em nosso país, em um paciente de 45 anos, devido ao consumo de um pó vendido como "ecstasy", apresentando como complicação uma síndrome coronariana aguda, com boa evolução posterior. A confirmação diagnóstica foi feita por sua detecção na urina por cromatografia gasosa acoplada à espectrometria de massa. A estrutura química da dibutilona e seu mecanismo de ação explicam as manifestações clínicas e complicações isquêmicas devido ao vasoespasmo coronariano. Na ausência de uma noção clara de exposição e dada a situação epidemiológica atual, a presença de quadro clínico compatível com droga estimulante e resultado negativo para cocaína e anfetaminas em estudos imunocromatográficos, deve levantar a suspeita de outros estimulantes sintéticos como as catinonas, como aconteceu neste caso.

Analytical quantification, intoxication case series, and pharmacological mechanism of action for N-ethylnorpentylone (N-ethylpentylone or ephylone).

Synthetic cathinones continue to proliferate in clandestine drug markets worldwide. N-ethylnorpentylone (also known as N-ethylpentylone or ephylone) is a popular emergent cathinone, yet little information is available about its toxicology and pharmacology. Here we characterize the analytical quantification, clinical presentation, and pharmacological mechanism of action for N-ethylnorpentylone. Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) was used to quantify N-ethylnorpentylone in blood obtained from human cases. Clinical features exhibited by the intoxicated individuals are described. The activity of N-ethylnorpentylone at plasma membrane transporters for dopamine (DAT), norepinephrine (NET) and 5-HT (SERT) was assessed using in vitro assays measuring uptake inhibition and evoked release of [3 H] neurotransmitters in rat brain synaptosomes. Our LC-MS/MS method assayed N-ethylnorpentylone concentrations with limits of detection and quantification of 1 and 5 ng/mL, respectively. Quantitation was linear from 5 to 500 ng/mL, and the method displayed specificity and reproducibility. Circulating concentrations of N-ethylnorpentylone ranged from 7 to 170 ng/mL in clinical cases, and the associated symptoms included palpitations, tachycardia, agitation, hallucinations, coma and death. N-Ethylnorpentylone was a potent inhibitor at DAT (IC50  = 37 nM), NET (IC50  = 105 nM) and SERT (IC50  = 383 nM) but displayed no transporter releasing activity. We present a validated method for quantifying N-ethylnorpentylone in human case work. The drug is a psychomotor stimulant capable of inducing serious cardiovascular and neurological side-effects which can be fatal. In vitro findings indicate that N-ethylnorpentylone exerts its effects by potent blockade of DAT and NET, thereby elevating extracellular levels of dopamine and norepinephrine in the brain and periphery.

The New Classes of Synthetic Illicit Drugs Can Significantly Harm the Brain: A Neuro Imaging Perspective with Full Review of MRI Findings.

Synthetic drugs contain substances that are pharmacologically similar to those found in traditional illicit drugs. Some of the most commonly abused synthetic drugs include synthetic marijuana, bath salts, ecstasy, N-bomb, methamphetamine and anabolic steroids. Many of them share the same chemical properties and physiologic responses with the drugs they mimic and may exaggerate the pathologic response in the brain leading to addiction. These drugs have detrimental (and often irreversible) effects on the brain and primarily affect the central nervous system by two mechanisms: 1) Neural hyper stimulation via increasing activation of certain neurotransmitters (norepinephrine, dopamine, and serotonin), 2) Cause significant reduction in CNS neural connectivity affecting various brain regions such as the basal ganglia, hippocampus, cerebellum, parietal lobe, and globus pallidus. Furthermore these drugs sometimes have severe, life-threatening adverse effects on the human body. A few structural MRI studies have been conducted in synthetic drug abusers to reveal the effects of these drugs on the brain parenchyma. This review article will describe the potential brain imaging findings in synthetic drug abusers as demonstrated by several case reports and the primary literature.

Brephedrone: A new psychoactive substance seized in Brazil.

At the beginning of 2015, sixty-two capsules containing red-brown crystals seized in a historical city in Brazil were sent to this forensic laboratory for drug testing analysis. The material was identified as being Brephedrone, a new psychoactive substance and a bromine synthetic cathinone that is related to serotonin transportation. This substance was analyzed by ATR-FTIR, GC-MS, LC-MS, 1H, 13C and 2D NMR. Brephedrone apprehensions have been previously reported in Finland, France and Spain. It was the first detection of this substance in the State of Minas Gerais. No reports or information regarding any other apprehension nor identification of Brephedrone in Brazil were known prior to the present case.

An approach to the new psychoactive drugs phenomenon / Una aproximación al fenómeno de las nuevas drogas psicoactivas

Abstract Background. The new psychoactive drugs (NPD) are those that represent a danger to public health and are not prohibited by conventions on international narcotics. The concept also includes new contexts and new routes of consumption as well as novel ways of distribution, notably Internet. The risks associated with NPD consumption are largely unknown to users and to health care providers. Objective. To integrate the existing evidence regarding the main NPD in terms of description, epidemiology, psychopharmacology, medical complications and psychoactive effects. Method. To review relevant and updated clinical information on NPD obtained from specialized books and indexed scientific journals (PubMed/Medline, Google Scholar, Scopus), as well as official documents edited by international organizations dedicated to the epidemiologic analysis of drug abuse and Internet websites and forums managed by psychoactive substance users. Results. Aspects of clinical and pharmacological interest are described comprehensively, together with epidemiological data and risks associated to the consumption of the most relevant NPD: synthetic cannabinoids, synthetic cathinones, NBOMe series, indoleamines, piperazines, hallucinogenic mushrooms (Psilocybe SP.), synthetic opioids, plant products (khat, kratom, Salvia divinorum, ayahuasca) and dissociative anesthetics. Discussion and conclusion. The emergence of the NPD is a phenomenon on the rise with important consequences for public health. Learning about new trends in drug consumption and its potential risks should be essential for the medical professional. New research is needed in order to understand the phenomenon of the NPD and its pharmacological, clinical and legal implications.

Resumen Antecedentes. Las nuevas drogas psicoactivas (NDP) son aquellas que, aun cuando representan un peligro para la salud pública, no están prohibidas por los acuerdos internacionales sobre narcóticos. La noción incluye también nuevos contextos de usos, nuevas formas de administración y nuevas vías de distribución, entre las que destaca Internet. Los riesgos asociados al consumo de NDP son en gran medida desconocidos por los usuarios y el personal de salud. Objetivo. Integrar la información existente sobre las principales NDP en cuanto a su descripción, psicofarmacología, epidemiología, efectos psicoactivos y complicaciones médicas descritas. Método. Revisión de la información actualizada de relevancia clínica sobre las NDP obtenida de libros especializados y revistas científicas indexadas (PubMed/Medline, Google Scholar, Scopus); de documentos oficiales de organismos internacionales dedicados a la epidemiología del consumo de drogas, y de portales y foros en Internet gestionados por usuarios de sustancias psicoactivas. Resultados. Se describen de manera detallada aspectos de interés clínico y farmacológico, así como datos epidemiológicos y riesgos asociados al consumo de las NDP más relevantes: cannabinoides sintéticos, catinonas sintéticas, serie de los NBOMe, indolaminas, piperazinas, hongos alucinógenos (Psilocybe sp.), opioides sintéticos, productos vegetales (khat, kratom, Salvia divinorum, ayahuasca) y anestésicos disociativos. Discusión y conclusión. El surgimiento de las NDP es un fenómeno en auge con importantes consecuencias en la salud pública. Se hace imprescindible para el profesional médico conocer las nuevas tendencias en el consumo y los riesgos potenciales del mismo. Son necesarias también nuevas investigaciones para comprender el fenómeno de las NDP y sus implicaciones farmacológicas, clínicas y legales.

Comparative pharmacological evaluation of the cathinone derivatives, mephedrone and methedrone, in mice.

Mephedrone and methedrone are cathinone-related compounds, which act as non-selective substrates for monoamine transporters, facilitating a neurotransmitter release. We compared the acute pharmacological effects of mephedrone and methedrone, attempting to further evaluate the action mechanisms of methedrone by responsibly and ethically using mice under approved procedures. The effects of both compounds were examined from 10 to 60 min, in a series of behavioral paradigms, namely open-field, plus-maze, hot-plate and tail suspension tests, whereas neurotransmitter brain tissue levels were determined ex vivo by HPLC. Separate groups were pre-treated with the dopamine (DA) antagonist haloperidol, or the serotonin (5-HT) synthesis inhibitor ρCPA, to further assess the mechanisms underlying methedrone effects. The compounds caused marked hyperlocomotion, displaying dissimilar stereotyped behavior, in an open-field arena. Mephedrone caused anxiolytic-like effects, while methedrone induced anxiogenic-like actions in the elevated plus-maze. Both compounds displayed thermal antinociception, with a reduced immobility time in the tail suspension model. Mephedrone triggered a 2- and 3-fold increment of dopamine and serotonin tissue levels, respectively, in the nucleus accumbens, with a 1.5-fold elevation of tissue dopamine in the frontal cortex. Methedrone caused a 2-fold increment of tissue dopamine in the nucleus accumbens and in the striatum, and a 1.5-fold increment of serotonin tissue levels in the hippocampus and striatum. In vivo methedrone effects were partially inhibited by a pre-treatment with haloperidol or ρCPA. Despite similar actions on locomotion, analgesia, and depression-like behavior, the acute administration of mephedrone and methedrone elicited divergent effects on anxiety-like behavior and stereotyped movements in mice, which might be related to the distinct modulation of brain tissue neurotransmitter levels.

Expressed sequence tag analysis of khat (Catha edulis) provides a putative molecular biochemical basis for the biosynthesis of phenylpropylamino alkaloids.

Khat (Catha edulis Forsk.) is a flowering perennial shrub cultivated for its neurostimulant properties resulting mainly from the occurrence of (S)-cathinone in young leaves. The biosynthesis of (S)-cathinone and the related phenylpropylamino alkaloids (1S,2S)-cathine and (1R,2S)-norephedrine is not well characterized in plants. We prepared a cDNA library from young khat leaves and sequenced 4,896 random clones, generating an expressed sequence tag (EST) library of 3,293 unigenes. Putative functions were assigned to > 98% of the ESTs, providing a key resource for gene discovery. Candidates potentially involved at various stages of phenylpropylamino alkaloid biosynthesis from L-phenylalanine to (1S,2S)-cathine were identified.

Expressed sequence tag analysis of khat (Catha edulis) provides a putative molecular biochemical basis for the biosynthesis of phenylpropylamino alkaloids

Khat (Catha edulis Forsk.) is a flowering perennial shrub cultivated for its neurostimulant properties resulting mainly from the occurrence of (S)-cathinone in young leaves. The biosynthesis of (S)-cathinone and the related phenylpropylamino alkaloids (1S,2S)-cathine and (1R,2S)-norephedrine is not well characterized in plants. We prepared a cDNA library from young khat leaves and sequenced 4,896 random clones, generating an expressed sequence tag (EST) library of 3,293 unigenes. Putative functions were assigned to > 98 percent of the ESTs, providing a key resource for gene discovery. Candidates potentially involved at various stages of phenylpropylamino alkaloid biosynthesis from L-phenylalanine to (1S,2S)-cathine were identified.