Caitlyn Norman

Designer benzodiazepines (DBZDs) are a class of new psychoactive substances (NPS) designed as legal alternatives to prescription BZDs. Bromazolam has been the most prevalent DBZD detected on the recreational market around the world; however, a new DBZD, ethylbromazolam (8-bromo-1-ethyl-6-phenyl-4 H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; also known as bromoethylazolam) has recently emerged. In this study, the emergence of ethylbromazolam in Canada, the UK, and Australia is reported based on analysis of samples from drug checking services and in Germany based on analysis of samples seized by customs and mail services. Since November 2024, ethylbromazolam has been increasingly detected with a concurrent decrease in bromazolam detections, suggesting that its emergence is likely in response to the international control of bromazolam on 3rd December 2024. Additionally, increased detections of other DBZDs, including desalkylgidazepam (bromonordiazepam) and clobromazolam (phenazolam) have been recently observed. The in vitro α ₁ β ₂ γ ₂ GABA _A receptor activity of ethylbromazolam was determined using an automated patch clamp assay. Ethylbromazolam was found to have similar in vitro GABA _A receptor activity as bromazolam (EC ₅₀ of 10.1 nM and 15.2 nM, respectively), indicating comparable pharmacological activity and potential for harm. The market should continue to be monitored closely as it continues to evolve in response to the control of bromazolam.

Background and Purpose: New synthetic cannabinoid receptor agonists (SCRAs) are associated with severe adverse effects, including unexpected psychiatric symptoms. These compounds are mainly active through their potent agonism on the cannabinoid receptors CB ₁ and CB ₂ . Recently, evidence of the direct interaction of some SCRAs with the serotonergic 5‐HT receptors has emerged. Experimental Approach: SCRAs were screened for in vitro 5‐HT _2A receptor activity using AequoScreen® intracellular Ca ²⁺ release and inositol monophosphate (IP ₁ ) formation assays. The pharmaco‐toxicological effects induced by the administration of the most effective compound in vitro (AB‐5′F‐BUTINACA) were investigated in male CD‐1 mice with specific focus on its interaction with CB ₁ and 5‐HT _2A receptors. Key Results: Many SCRAs with a valinamide (AB) or tert ‐leucinamide (ADB) head moiety had in vitro activity at the 5‐HT _2A receptor at high concentrations with AB‐5′F‐BUTINACA and ADB‐BUTINACA amongst the most efficacious. In vivo studies showed that AB‐5′F‐BUTINACA caused neurological changes, sensorimotor alteration, antinociception, hypothermia, reduction in breath rate and hypolocomotion in mice. The neurological, sensorimotor, and thermal antinociceptive effects were mediated by CB ₁ and 5‐HT _2A receptors, as they were prevented by pretreatment with the selective CB ₁ and 5‐HT _2A receptor antagonists (NESS0327 and MDL100907, respectively). Conclusion and Implications: This study demonstrated for the first time that SCRAs with a valinamide or tert ‐leucinamide head moiety activate the 5‐HT _2A receptor. Agonism of the compound AB‐5′F‐BUTINACA at the 5‐HT _2A receptors resulted in in vivo effects. Our data revealed potential risks related to SCRA consumption that could exacerbate unexpected psychiatric conditions.

Although widely prescribed globally, benzodiazepines (BZDs) are commonly misused and involved in a high rate of drug-related deaths worldwide. Novel chemically similar compounds, designer BZDs (DBZDs), have emerged on the recreational market with little information available on their pharmacodynamic effects. An in vitro α ₁ β ₂ γ ₂ GABA _A assay was used to determine the activity and explore structure-activity relationships of 42 DBZDs plus 11 prescription BZDs. Their interaction with the BZD α ⁺ /γ ₂ ^– interface was explored using the BZD antagonist flumazenil. Most of the D/BZDs were positive allosteric modulators, but 4’-chlorodiazepam and 4’-fluorodiazepam were negative allosteric modulators, although the activity was not antagonized by flumazenil. All metabolites tested were active. This data informs the interpretation of intoxications, harm reduction measures, and drugs legislation for DBZDs; demonstrates the importance of testing new drugs as they emerge on the recreational market; and offers insights into how BZD chemical modifications alter GABA _A pharmacodynamics.

Synthetic cannabinoid receptor agonists (SCRAs) is a large group of structurally diverse designer drugs (analogues of controlled substances) associated with intense and sometimes fatal intoxication. Cardiac symptoms including tachycardia and arrhythmia are common consequences of SCRA consumption. However, little is known about the mechanisms through which SCRAs may perturb cardiac rhythm. Here we used electrophysiological techniques to screen 36 SCRAs on two ion channels responsible for cardiomyocyte repolarization, hERG (also called KV11.1) and KV7.1/KCNE1. We report that the majority of tested SCRAs inhibited hERG, primarily by reducing channel conductance, and some also inhibited KV7.1/KCNE1. In silico data suggests that SCRAs use a known drug binding site in the pore of the hERG channel, shared by established hERG blockers like astemizole, where a planar SCRA molecule lays perpendicular to the ion conducting pathway. Experimental and in silico data identify SCRA structural features associated with prominent inhibitory effects on hERG, with chemical moieties allowing bond formation and/or the ability to fit into the vestibule being important. Structure-activity-relationships for SCRA effects on hERG, Kv7.1/KCNE1 and the cannabinoid receptor 1 (CB1) varied, demonstrating the importance of assessing SCRA effects on multiple potential targets. In conclusion, we found SCRAs to be inhibitors of cardiac voltage-gated potassium channels important for cardiomyocyte repolarization. These results offer mechanistic insight into potentially detrimental SCRA effects on the heart and highlight the urgency of more extensive investigation of SCRAs on cardiac function.

An emerging new trend reveals that potent synthetic cannabinoid receptor agonists (SCRAs) that were prevalent prior to the Chinese class-wide ban in July 2021 have remained on the market or re-emerged. This is likely because of a new production route where unscheduled tail-less precursors are converted into the desired controlled SCRA via a one-step synthesis. This study reports the discovery of a clandestine production site in Switzerland for this synthesis. Controlled recreations of the synthesis of ADB-BUTINACA (also known as ADB-BINACA or ADMB-BUTINACA) and MDMB-4en-PINACA based on a recipe recovered from the Swiss production site were undertaken. The recreations yielded oils or waxy products with chromatographic purities ranging from 39.3 to 93.4% with the precursors (ADB-INACA or MDMB-INACA) remaining in all products. Mixtures of a precursor (e.g., MDMB-INACA, ADB-INACA, AB-INACA, MDMB-ICA, ADB-IATA, MDMB-5’Me-INACA) with corresponding final SCRA(s) (e.g., MDMB-4en-PINACA, 4F-MDMB-BUTINACA, MDMB-BUTINACA, MDMB-FUBINACA, ADB-BUTINACA, ADB-4en-PINACA, AB-CHMINACA, 4F-MDMB-BUTICA, ADB-FUBIATA) were found in 28 forensic casework samples in the US received between February 2023 and February 2024 and 87 samples seized from the Scottish prisons between February 2023 and July 2024. In vitro CB₁ receptor activity of the precursors, synthesized products, and mixtures of precursors and final SCRAs were examined using β-arrestin 2 recruitment and intracellular calcium release assays. The precursors were much less potent than the final SCRAs and the concentration of the final SCRA dictated the activity of mixtures. This study raises awareness of the emergence of a new production method for SCRAs as observed in multiple countries.

Synthetic cannabinoid receptor agonists (SCRAs) are a group of new psychoactive substances (NPS) that bind to and activate the cannabinoid 1 and 2 (CB₁ and CB₂) receptors. Following the introduction of SCRA analog controls in China in July 2021, new SCRAs with a bromine at the 5 position on the indazole core emerged on the recreational drug market. In this study, the in vitro CB₁ receptor activity of 19 different SCRAs with a halogenated indazole core was systematically examined using a AequoScreen^® CB₁ assay alongside four available non-halogenated analogs, ADB-BUTINACA, MDMB-BUTINACA, MDMB-4en-PINACA, and MDMB-INACA, and the newly emerged MDMB-5′Me-INACA (MDMB-5-methyl-INACA). Of SCRAs with a substitution at the 5 position on the indazole core, analogs with a fluorine had the lowest EC₅₀ values. For SCRAs with a methyl ester head moiety (MMB or MDMB), chlorinated analogs had the next lowest EC₅₀ values followed by brominated analogs; however, the opposite relationship was observed for SCRAs with an amide head moiety (AB or ADB). All halogenated compounds had similar potency to their non-halogenated analog, except the brominated tert-leucine methyl ester SCRAs, which had significantly reduced potency. Of the different head moieties, tert-leucine methyl ester SCRAs were the most potent, followed by tert-leucinamide, valinamide, and valine methyl ester. The potencies of the SCRAs with pentyl (ADB-5′Br-PINACA/ADMB-5′Br-PINACA/ADB-P-5′Br-INACA), pent-4-enyl (ADB-4en-5′Br-PINACA/ADMB-4en-5′Br-PINACA/ADB-4en-P-5′Br-INACA and MDMB-4en-5′Br-PINACA/MDMB-4en-P-5′Br-INACA), and butyl tails (ADB-5′Br-BUTINACA/ADMB-5′Br-BUTINACA/ADB-B-5′Br-INACA) were not significantly different, whereas fluorobenzyl (ADB-5′Br-FUBINACA), tail-less (ADB-5′Br-INACA), and decyl tail (ADB-5′Br-DECINACA/ADMB-5′Br-DECINACA/ADB-D-5′Br-INACA) analogs were significantly less potent.

Novel psychoactive substances (NPS) pose one of the greatest challenges across the illicit drug landscape. They can be highly potent, and coupled with rapid changes in structure, tracking and identifying these drugs is difficult and presents users with a "Russian roulette" if used. Benzodiazepines, synthetic opioids, synthetic cannabinoids, and synthetic cathinones account for the majority of NPS-related deaths and harm. Detecting these drugs with existing field-portable technologies is challenging and has hampered the development of community harm reduction services and interventions. Herein, we demonstrate that hybridizing fluorescence and reflectance spectroscopies can accurately identify NPS and provide concentration information with a focus on benzodiazepines and nitazenes. The discrimination is achieved through a deep learning algorithm trained on a library of preprocessed spectral data. We demonstrate the potential for these measurements to be made using a low-cost, portable device that requires minimal user training. Using this device, we demonstrate the discrimination of 11 benzodiazepines from "street" tablets that include bulking agents and other excipients. We show the detection of complex mixtures of multiple drugs, with the key example of nitazene + benzodiazepine (metonitazene + bromazolam), fentanyl + xylazine, and heroin + nitazene (etonitazene) combinations. These samples represent current drug trends and are associated with drug-related deaths. When combined with the implementation of detection technology in a portable device, these data point to the immediate potential to support harm reduction work in community-based settings. Finally, we demonstrate that the approach may be generalized to other drug classes outside NPS discrimination.

After the Scottish Prison Service (SPS) introduced mail photocopying procedures in December 2021, a shift in smuggling methods was observed for synthetic cannabinoid receptor agonists (SCRAs) and other new psychoactive substances (NPS) from drug‐infused papers back to traditional sample matrices (e.g., tablets and powders), although new matrices also emerged. This study reports on waxy‐ or putty‐like materials as a novel drug preparation for SCRAs and other drugs seized from UK prisons. In 2023, 22 of these new preparations were seized from Scottish prisons, with eight found in sealed vape pods. The materials were positive for SCRAs, phytocannabinoids, novel benzodiazepines, and/or gabapentinoids. Additionally, 11 preparations were seized from an English prison, all containing the SCRAs MDMB‐4en‐PINACA and MDMB‐INACA. MDMB‐INACA was pharmacologically characterized using in vitro CB₁ and CB₂ bioassays, revealing moderate efficacy but low potency at CB₁. Furthermore, the in vitro CB₁ bioassay was also used to evaluate the CB₁ activating potential of extracts from eight seized samples. Six of these showed high CB₁ activity, whereas the samples lacking SCRAs or containing only MDMB‐INACA showed no or only weak CB₁ activity, respectively. Lastly, applying the bioassay as an activity‐based “untargeted” screening method effectively identified the presence of SCRAs in one waxy preparation, which was initially not detected by gas chromatography–mass spectrometry (GC‐MS). This underscores the effectiveness of the bioassay for evaluating these new waxy‐ or putty‐like materials for the presence of SCRAs.

Novel Psychoactive Substances (NPS) pose one of the greatest challenges across the illicit drug landscape. They can be highly potent, and coupled with rapid changes in structure, tracking and identifying these drugs is difficult, and presents users with a ‘Russian roulette’ if used. Benzodiazepines, synthetic opioids, synthetic cannabinoids and synthetic cathi-nones account for the majority of NPS related deaths and harm. Detecting these drugs with existing field-portable technol-ogies is challenging and has hampered the development of community harm reduction services and interventions. Herein, we demonstrate that hybridizing fluorescence and reflectance spectroscopies can accurately identify NPS and provide concentration information, with a focus on benzodiazepines and nitazenes. The discrimination is achieved through a deep learning algorithm trained on a library of pre-processed spectral data. We demonstrate the potential for these measure-ments to be made using a low-cost, portable device that requires minimal user training. Using this device, we demon-strate the discrimination of 11 benzodiazepines and from ‘street’ tablets that include bulking agents and other excipients. We show the detection of complex mixtures of multiple drugs, with the key example of nitazene + benzodiazepine (metonitazene + bromazolam), fentanyl + xylazine and heroin + nitazene (etonitazene) combinations. These samples represent current drug trends and associated with drug related deaths. When combined with the implementation of detection technology in a portable device, these data point to the immediate potential to support harm reduction work in community-based settings. Finally, we demonstrate that the approach may be more broadly generalized to other drug classes outside of NPS discrimination.

Anabolic‐androgenic steroids (AASs) are a subclassification of image performance enhancing drugs (IPEDs). While AAS use is most prevalent among people in athletics, there is also high lifetime prevalence of AAS use among prisoners. This study reports the qualitative detection of AASs in seized samples from the Scottish prisons from 2019–2023. Additionally, methods were developed for the quantitative analysis of AASs using gas chromatography–mass spectrometry (GC–MS) and applied to 61 samples of tablets or powders seized from Scottish prisons between July 2022 and July 2023. Since 2022, there has been an increase in AAS detections in the Scottish prisons. Oxymetholone was the most prevalent AAS, followed by metandienone (methandrostenolone, methandienone), methyltestosterone, oxandrolone, mestanolone (methylandrostanolone), stanozolol, and androstenedione. Multiple AASs were found in 21 samples and 10 samples contained other drugs, including amitriptyline, sertraline, zopiclone, mirtazapine, sildenafil, etizolam, Δ⁹‐tetrahydrocannabinol, and the synthetic cannabinoid MDMB‐INACA. Most AAS samples were tablets (77.0%), although they were also detected in powders, herbal material, e‐cigarettes, and a fragmented soap bar‐type sample. There was a large variation in the concentration of AASs in the tablets and powders seized from the Scottish prisons, demonstrating AASs are another highly variable component of the polydrug use situation in prisons, the effects of which need to be examined further.