Contexte : L’insomnie chronique affecte un nombre important de personnes âgées; environ 34 % de ces derniers se plaignent d’une difficulté persistante à s’endormir. Beaucoup d’aînés se tournent vers la prise d’hypnotiques pour traiter leur insomnie. Plus précisément, vers une certaine classe d’hypnotiques qu’on nomme les benzodiazépines (BZD). Or, la prise de benzodiazépines est associée à plusieurs impacts négatifs sur la santé chez les personnes âgées (fracture de la hanche, mortalité, cancers et déclin cognitif). Par ailleurs, la thérapie cognitivo-comportementale pour insomnie (TCCI) est considérée comme un traitement de première ligne pour le trouble d’insomnie. De plus, elle améliore plusieurs variables de sommeil (latence d’endormissement, durée totale de sommeil, durée d’éveil et efficacité de sommeil). Objectif : L’objectif de cette étude pilote a pour but d’évaluer l’effet de la TCCI sur le sommeil au cours d’un sevrage structuré de benzodiazépines chez des participants âgés de 60 ans et plus. Méthodes : Trente-six (36) participants dont vingt-quatre (24) femmes et douze (12) hommes d’une moyenne d’âge de 69,5 ans (60-90 ans) souffrant d’insomnie chronique et prenant des BZD sur une durée moyenne de 15 ans (1-44 ans) ont effectué une évaluation de leur sommeil approfondie à travers des agendas de sommeil, des actigraphies et des enregistrements polysomnographiques, avant d’être répartis aléatoirement en deux groupes : le groupe thérapie (n = 19) avec un programme de sevrage sur seize semaines associé à huit sessions de TCCI, et un groupe témoin (n = 17) avec un programme de sevrage associé à la remise d’une brochure explicative des étapes d’un sevrage structuré. Les mêmes évaluations de sommeil ont été répétées à l’issue du sevrage des deux groupes. Des ANCOVAs à mesures répétées et des tests de χ2 de Pearson ont évalué l’effet de l’intervention thérapeutique sur les variables de sommeil et le succès de sevrage. 4 Résultats et conclusion : 65 % de la totalité des participants sont arrivés à une cessation complète de leurs BZD (73,68 % des participants du groupe thérapie et 56,25 % du groupe témoin). Selon les valeurs obtenues sur les agendas de sommeil, le groupe thérapie s’est amélioré davantage que le groupe témoin par la réduction de la latence d’endormissement (F = 6,15, ddl = 1, p = 0,02), de la durée d’éveil (F = 10,96, ddl = 1, p < 0,001) et l’augmentation de l’efficacité de sommeil (F = 4,84, ddl = 1, p = 0,03). De surcroît, le groupe thérapie montre une tendance vers une amélioration au niveau de la qualité de sommeil, tel que défini par le questionnaire d’IQSP1 (F = 3,46, ddl = 1, p = 0,06). Ces résultats préliminaires suggèrent un effet bénéfique de la TCCI sur le sommeil, dans le cadre d’un sevrage de benzodiazépines. / Background: Chronic insomnia affects a significant number of the elderly population, with approximately 34% of older adults complaining of persistent difficulty falling asleep. Many of these older adults choose to use hypnotics to treat their insomnia. To be more specific, they tend to use a certain class of hypnotics called benzodiazepines (BZD). However, benzodiazepine use is correlated with several negative health impacts in the elderly (hip fracture, increased risk of mortality, cancer and cognitive decline). In addition, Cognitive behavioral therapy for insomnia (CBTI) is considered the gold standard and the first-line treatment for primary insomnia disorder. In addition, CBTI improves several sleep variables (sleep latency, total sleep time, waking duration and sleep efficiency). Objective: The objective of this pilot study is to evaluate the effect of CBTI on sleep during a structured benzodiazepine withdrawal in participants who are 60 years of age and older. Methods: Thirty-six (36) participants, twenty-four (24) of whom are female and twelve (12) of whom are male with a mean age of 69.5 years (60-90 years) with chronic insomnia and taking BZDs over an average time of 15 years (1-44 years) completed a comprehensive sleep assessment through sleep diaries, actigraphy, and polysomnographic recordings, before being randomly divided into two groups: the therapy group (n = 19) with a 16-week weaning program associated with eight CBTI sessions, and a control group (n = 17) with a weaning program associated with providing an explanatory brochure of the steps of structured weaning. The same sleep assessments were repeated after weaning of both groups. Repeated measures ANCOVAs and Pearson’s χ2 tests evaluated the effect of therapeutic intervention on sleep variables and withdrawal success. 6 Results and conclusion: 65% of all participants reached a complete cessation of their BZDs (73.68% of participants in the therapy group and 56.25% of the control group). According to the values obtained from sleep diaries, the therapy group improved more than the control group in terms of a reduction in the sleep onset latency (F = 6.15, df = 1, p = 0.02), wake after sleep onset (WASO) (F = 10.96, df = 1, p < 0.001) and an increase in the sleep efficiency (F = 4.84, df = 1, p = 0.03). In addition, the therapy group shows a trend toward improvement in sleep quality, as reported by the questionnaire of PSQI2 (F = 3.46, df = 1, p = 0.06). These preliminary results suggest a beneficial effect of CBTI on sleep as part of a program of structured withdrawal from benzodiazepines.

More than 20 years after its discovery HIV protease still remains one of the primary targets in HIV treatment. Currently there are 9 approved protease inhibitors on the market. However, due to immense replication rate and the high error prone nature of reverse transcriptase, resistance to each of them has already been described. Therefore, the search for new protease inhibitors with different binding mode is still active. A novel type of protease inhibitors (1, 4-benzodiazepine analogs) was recently discovered in our laboratory. Even though this new class of inhibitors is highly potent (Ki' in range of 10-9 ), it also has several undesirable qualities, such as low solubility and a high number of stereogenic centers. Primary objective of this study was to try to prepare more soluble compounds with lower number of possible stereoisomers, enzymologically characterize its binding to the wild-type and mutated HIV protease and to determine its structure in the complex with the enzyme. A small library of 1, 4-benzodiazepine inhibitors of HIV protease was synthesized and fully characterized using NMR spectroscopy and mass spectroscopy. The number of stereogenic centers was successfully reduced from 4 to 2 without loosing activity of the inhibitor. The improvement in solubility was always associated with a...

Introduction: Post-mortem whole blood samples differ greatly in quality, lipemia is one cause of concern in toxicological analyses. Around 4 % of all samples sent to RMV are given a notation of lipemic content. The aim of the thesis was to study the effects of lipemia on the quantification of 14 benzodiazepines and 5 similar sedative and antianxiety drugs as well as evaluate the pre-analytical process aiming to reduce the effects of lipemia.  Methods: Blood samples were simulated with bovine blood, analyte spiking, and lipid spiking with either the nutrition emulsion Intralipid or with a mixture of post-mortem lipids from authentic samples. The outset was the by RMV currently used LLE method followed by UPLC- MS/MS and the extraction method was altered and evaluated. Matrix effects were also studied.  Results: Lipemia were found to be a great interference when quantifying benzodiazepines. For most analytes, internal standard could compensate for the loss of analyte but there was a problem with analytes not having their own IS. The 7-amino-compounds were greatly affected by lipemia and propiomazine and dihydropropiomazine showed extreme losses. Equilibration of IS did not result in similar loss as analyte. Dilution of sample reduced losses caused by lipemic content. SPE resulted in extracts free from lipids and high yields but there were analyte losses similar to LLE. No matrix effects from the lipids were found. Samples spiked with Intralipid gave poorer analyte yields than those spiked with post-mortem lipids.  Conclusion: Dilution is the most successful method to reduce pre-analytical matrix effects as long as the concentration is not so low that it risks getting lower than the analytical limits when doing so. Not homogenising samples before sampling is giving incorrect results. SPE could, if optimised for the analyte retention and elution, remove lipids from samples and obtain accurate analyte concentrations. Pooling lipids from post-mortem samples is a possible method for simulating lipemic whole blood. Intralipid and the PM-mix gave the same indications, but to different extents. Further studies where the ability to mimic authentic lipids are needed for both Intralipid and PM-mix.

No description available.

Poly-(ADP-ribsoyl) Polymerases (PARPs) are a superfamily of enzymes comprised of 17 known isoforms. PARP inhibitors (PARPi) have shown success in clinical trials for the treatment of homologous recombination-deficient cancers. Though proven effective initially, tumors treated with PARPi eventually develop resistance. Combinatorial therapeutics targeting PARP and other pathways that may re-sensitize tumors to PARP inhibition, including PI3K/AKT/mTor pathway, and cell-cycle checkpoints (such as CDKs, CHK, and Wee) are being tested. In this context, the synthetic lethality of cyclin-dependent kinase 1 (CDK1) and PARP1 is known. Evaluation of PARP1 and CDK1 pharmacophores led to the development of the tetrahydro-arylazepinone (TAAP) scaffold as a potential dual PARP1/CDK1 inhibitor. We screened a handful of TAAP analogs against PARP1 in a cell-free assay that identified the low micromolar PARP1 inhibitor 1,2,3,4-tetrahydro-5H-benzo[e][1,4]-diazepin-5-one (TBAP), which served as the lead compound. The analogous 1,2,3,4-tetrahydro-5H-pyrido[2,3-e][1,4]-diazepin-5-one (TPAP) series showed a similar bioactivity profile. Satisfyingly, the N1-benzyl TPAP analogue showed activity in the low nanomolar range. The TAAP series (i.e., 6/7-membered scaffold) unfortunately lacked CDK1 inhibitory activity. Finally, many PARPi's show poor isoform-selectivity. The development of isoform-selective PARPi can clarify the specific function of each PARP isoform and may reduce the adverse side effects shown by PARPi. A handful of TAAP analogs were screened against 13 PARP isoforms, where some compounds demonstrated exquisite PARP1/2 selectivity. Concurrently, we discovered an inhibitor for PARP11, an isoform that lacks any known synthetic ligand. Future directions are suggested towards fine-tuning the structure-activity relationship of TAAP-isoform selective PARPi as well as developing a dual PARP1/CDK1 inhibitor. / Master of Science

Yes / The number of benzodiazepines appearing as new psychoactive substances (NPS) is continually increasing. Information about the pharmacological parameters of these compounds is required to fully understand their potential effects and harms. One parameter that has yet to be described is the blood-to-plasma ratio. Knowledge of the pharmacodynamics of designer benzodiazepines is also important, and the use of quantitative structure–activity relationship (QSAR) modelling provides a fast and inexpensive method of predicting binding affinity to the GABAA receptor. Methods: In this work, the blood-to-plasma ratios for six designer benzodiazepines (deschloroetizolam, diclazepam, etizolam, meclonazepam, phenazepam, and pyrazolam) were determined. A previously developed QSAR model was used to predict the binding affinity of nine designer benzodiazepines that have recently appeared. Results: Blood-to-plasma values ranged from 0.57 for phenazepam to 1.18 to pyrazolam. Four designer benzodiazepines appearing since 2017 (fluclotizolam, difludiazepam, flualprazolam, and clobromazolam) had predicted binding affinities to the GABAA receptor that were greater than previously predicted binding affinities for other designer benzodiazepines. Conclusions: This work highlights the diverse nature of the designer benzodiazepines and adds to our understanding of their pharmacology. The greater predicted binding affinities are a potential indication of the increasing potency of designer benzodiazepines appearing on the illicit drugs market. / Engineering and Physical Sciences Research Council. / Research Development Fund Publication Prize Award winner, Feb 2022.

O protocolo de teste/reteste no labirinto em cruz elevado (LCE) mostra que a experiência prévia no labirinto produz alterações duradouras nas respostas comportamentais de roedores. Nesse contexto, ratos submetidos ao LCE pela primeira vez apresentam um aumento característico na exploração dos braços abertos e uma redução dos comportamentos de avaliação de risco após a administração de drogas ansiolíticas. Na reexposição ao labirinto, porém, essas drogas tornam-se ineficazes em alterar as medidas tradicionais do LCE. Esse fenômeno foi inicialmente observado com o benzodiazepínico clordiazepóxido e referido como one-trial tolerance (tolerância de um ensaio OTT). A proposta do presente estudo é compreender a OTT por meio do exame dos fatores hormonais, cognitivos e neuroanatômicos envolvidos nesse fenômeno. A administração sistêmica do benzodiazepínico midazolam ou de metirapona, um bloqueador da síntese de glicocorticóides, reduziu a frequência dos comportamentos de avaliação de risco e dos níveis plasmáticos de corticosterona quando injetados antes das sessões teste ou reteste. Além disso, a reexposição de ratos ao LCE foi caracterizada por uma avaliação de risco mais proeminente, de acordo com a análise fatorial, e pela ativação de estruturas límbicas envolvidas com aspectos cognitivos do medo, como a região ventral do córtex pré-frontal medial (CPFm) e a amígdala, mostrada por meio da distribuição da proteína Fos. Midazolam administrado antes da primeira exposição ao LCE produziu uma redução significativa do número de neurônios Fos-positivos no córtex cingulado anterior, área 1 (Cg1) e nos núcleos anterior e pré-mamilar dorsal do hipotálamo. Por outro lado, midazolam causou uma redução no número de neurônios Fos-positivos no CPFm, amígdala, núcleo dorsomedial do hipotálamo e núcleos da rafe em ratos reexpostos ao LCE. Cg1 foi a única estrutura-alvo do benzodiazepínico em ambas as sessões. Resultados comportamentais similares aos produzidos pelo tratamento sistêmico foram obtidos com infusões de midazolam intra-Cg1. Esses resultados apontam para um papel crucial dos comportamentos de avaliação de risco no desenvolvimento da OTT e indicam o Cg1 como um importante sítio de ação ansiolítica dos benzodiazepínicos em roedores. / The elevated plus maze (EPM) test/retest protocol has shown that prior experience to the maze produces enduring changes in behavioral responses of rodents. In this context, rats submitted for the first time to the EPM display a characteristic increase in open arm exploration and reduced risk assessment behaviors after the administration of anxiolytic drugs. Upon re-exposure to the maze, however, these drugs become unable to change the traditional measures of the EPM. This phenomenon was initially observed with the benzodiazepine chlordiazepoxide and referred to as one-trial tolerance (OTT). The purpose of the present study is to understand the OTT through the exam of the hormonal, cognitive and neuroanatomical factors involved in this phenomenon. The systemic administration of the benzodiazepine midazolam or metyrapone, a glucocorticoids synthesis blocker, reduced the frequency of risk assessment behaviors and the corticosterone levels when injected before the test or retest sessions. Moreover, the re-exposure of rats to the EPM was characterized by more prominent risk assessment behaviors, according to the factor analysis, and by activation of limbic structures involved with cognitive aspects of fear, such as the ventral regions of the medial prefrontal cortex (mPFC) and amygdala, as shown through the distribution of the Fos protein. Midazolam injected before the first exposure to the EPM produced a significant decrease in the number of Fos-positive neurons in the anterior cingulate cortex, area 1 (Cg1), anterior and dorsal premammillary nuclei of hypothalamus. On the other hand, midazolam caused a decrease in the number of Fos-positive neurons in the mPFC, amygdala, dorsomedial nucleus of hypothalamus and raphe nuclei in rats re-exposed to the EPM. Cg1 was the only structure targeted by the benzodiazepine in both sessions. Behavioral results similar to those produced by systemic treatment were obtained with intra-Cg1 infusions of midazolam. These results point to a crucial role of the risk assessment behaviors in the development of the OTT and indicate the Cg1 as an important locus for the anxiolytic-like action of benzodiazepines in rodents.

Psychotropic medication use is widely implicated as a risk factor for injuries, and it is believed that the adverse effect profiles of these medications are exacerbated by the consumption of alcohol. The objectives of this study are (a) to examine the associations between the use of specific classes of psychotropic medications and injuries among elderly participants of the National Population Health Survey (NPHS), and (b) to determine whether and how associations between psychotropic medications and injuries are modified by the consumption of alcohol. Data from Cycles 1 (1994/95), 2 (1996/97), and 3 (1998/99) of the NPHS household longitudinal file were used in this study, selecting community-dwelling participants aged 65 years of age and older in 1994/95. Among antidepressant medications, the magnitude of the risk of injuries was higher for users of tricyclic derivatives (OR=1.4; 95%CI: 0.7 – 2.9) than SSRIs (OR=0.3; 95%CI: 0.1 – 1.0). Benzodiazepine use for any indication increased the risk of injuries, but that effect was not consistent across indications. The use of benzodiazepine antianxiety medications resulted in an increased risk of injuries (OR=2.0; 95%CI: 1.3 – 3.1), but there were no significant effects on the injury risk among benzodiazepine hypnotic and sedative users (OR=0.8; 95%CI: 0.4 – 1.7). Results pertaining to the second objective of this study raised as many questions as they resolved. Alcohol consumption decreased the odds of injury among hypnotic and sedative users, but otherwise, no consistent results were observed. Findings from this study underscore the importance of identifying appropriate alcohol measures for research among elderly populations. They also stress the need to separately consider the impact of different classes of psychotropic medications on injuries (tricyclic antidepressants separate from SSRI antidepressants and antianxiety benzodiazepines separate from hypnotic and sedative benzodiazepines).

Psychotropic medication use is widely implicated as a risk factor for injuries, and it is believed that the adverse effect profiles of these medications are exacerbated by the consumption of alcohol. The objectives of this study are (a) to examine the associations between the use of specific classes of psychotropic medications and injuries among elderly participants of the National Population Health Survey (NPHS), and (b) to determine whether and how associations between psychotropic medications and injuries are modified by the consumption of alcohol. Data from Cycles 1 (1994/95), 2 (1996/97), and 3 (1998/99) of the NPHS household longitudinal file were used in this study, selecting community-dwelling participants aged 65 years of age and older in 1994/95. Among antidepressant medications, the magnitude of the risk of injuries was higher for users of tricyclic derivatives (OR=1.4; 95%CI: 0.7 – 2.9) than SSRIs (OR=0.3; 95%CI: 0.1 – 1.0). Benzodiazepine use for any indication increased the risk of injuries, but that effect was not consistent across indications. The use of benzodiazepine antianxiety medications resulted in an increased risk of injuries (OR=2.0; 95%CI: 1.3 – 3.1), but there were no significant effects on the injury risk among benzodiazepine hypnotic and sedative users (OR=0.8; 95%CI: 0.4 – 1.7). Results pertaining to the second objective of this study raised as many questions as they resolved. Alcohol consumption decreased the odds of injury among hypnotic and sedative users, but otherwise, no consistent results were observed. Findings from this study underscore the importance of identifying appropriate alcohol measures for research among elderly populations. They also stress the need to separately consider the impact of different classes of psychotropic medications on injuries (tricyclic antidepressants separate from SSRI antidepressants and antianxiety benzodiazepines separate from hypnotic and sedative benzodiazepines).

For patients afflicted with symptoms of anxiety and insomnia, benzodiazepines are generally a safe and effective short-term pharmacological treatment option. Although considered safer than other sedative-hypnotic medications, substantial concern exists regarding the addictive nature and abuse potential of benzodiazepines along with potentially inappropriate prescribing and utilization in clinically vulnerable populations. These medication misadventures can have a significant impact on public health. Examples of medication misadventures as they pertain to benzodiazepines include the prescribing and use in clinically vulnerable populations for whom they are contraindicated or their efficacy has not been evaluated, the development of tolerance or addiction, abuse of the medication, and the manifestation of negative health outcomes including cognitive impairment, withdrawal symptoms upon discontinuation, or the reoccurrence of a preexisting substance use disorder. In order to better understand medication misadventures associated with benzodiazepines retrospective analyses using populations extracted from large health claims databases are employed. To understand how benzodiazepine use may lead to adverse events causing patient harm, the risk of exacerbations in benzodiazepine users diagnosed with chronic obstructive pulmonary disease was estimated. The inherent risk of benzodiazepine addiction and abuse was estimated in an HIV-infected population, a population with a high prevalence of substance use disorders. This risk was estimated by first determining whether HIV-infected individuals are more likely to have any benzodiazepine use compared to their uninfected counterparts, and secondly, by examining the association between HIV-infection and potentially problematic benzodiazepine use. Finally, in an effort to mitigate unexpected and undesirable consequences to public health associated with the prescription drug abuse epidemic in the US, states have implemented prescription drug monitoring programs (PDMPs) to track the prescribing and dispensing of controlled substance medications. The effect of these programs on benzodiazepine dispensing is evaluated on a state and national level. Findings will provide healthcare professionals a better understanding regarding the risk of medication misadventures involving benzodiazepines when evaluating their appropriateness in patients with anxiety, depression, and insomnia. Additionally, policymakers will understand the implications of PDMPs on the dispensing of benzodiazepines as they become a more widely used tool to combat prescription drug abuse and diversion.