Trace Amine-Associated Receptor 1 Contributes to Diverse Functional Actions of O-Phenyl-Iodotyramine in Mice but Not to the Effects of Monoamine-Based Antidepressants.

Trace Amine-Associated Receptor 1 (TAAR1) is a potential target for the treatment of depression and other CNS disorders. However, the precise functional roles of TAAR1 to the actions of clinically used antidepressants remains unclear. Herein, we addressed these issues employing the TAAR1 agonist, o-phenyl-iodotyramine (o-PIT), together with TAAR1-knockout (KO) mice. Irrespective of genotype, systemic administration of o-PIT led to a similar increase in mouse brain concentrations. Consistent with the observation of a high density of TAAR1 in the medial preoptic area, o-PIT-induced hypothermia was significantly reduced in TAAR1-KO mice. Furthermore, the inhibition of a prepulse inhibition response by o-PIT, as well as its induction of striatal tyrosine hydroxylase phosphorylation and elevation of extracellular DA in prefrontal cortex, were all reduced in TAAR1-KO compared to wildtype mice. O-PIT was active in both forced-swim and marble-burying tests, and its effects were significantly blunted in TAAR1-KO mice. Conversely, the actions on behaviour and prefrontal cortex dialysis of a broad suite of clinically used antidepressants were unaffected in TAAR1-KO mice. In conclusion, o-PIT is a useful tool for exploring the hypothermic and other functional antidepressant roles of TAAR1. By contrast, clinically used antidepressants do not require TAAR1 for expression of their antidepressant properties.

MCH-R1 Antagonist GPS18169, a Pseudopeptide, Is a Peripheral Anti-Obesity Agent in Mice.

Melanin-concentrating hormone (MCH) is a 19 amino acid long peptide found in the brain of animals, including fishes, batrachians, and mammals. MCH is implicated in appetite and/or energy homeostasis. Antagonists at its receptor (MCH-R1) could be major tools (or ultimately drugs) to understand the mechanism of MCH action and to fight the obesity syndrome that is a worldwide societal health problem. Ever since the deorphanisation of the MCH receptor, we cloned, expressed, and characterized the receptor MCH-R1 and started a vast medicinal chemistry program aiming at the discovery of such usable compounds. In the present final work, we describe GPS18169, a pseudopeptide antagonist at the MCH-R1 receptor with an affinity in the nanomolar range and a Ki for its antagonistic effect in the 20 picomolar range. Its metabolic stability is rather ameliorated compared to its initial parent compound, the antagonist S38151. We tested it in an in vivo experiment using high diet mice. GPS18169 was found to be active in limiting the accumulation of adipose tissues and, correlatively, we observed a normalization of the insulin level in the treated animals, while no change in food or water consumption was observed.

S29434, a Quinone Reductase 2 Inhibitor: Main Biochemical and Cellular Characterization.

Quinone reductase 2 (QR2, E.C. 1.10.5.1) is an enzyme with a feature that has attracted attention for several decades: in standard conditions, instead of recognizing NAD(P)H as an electron donor, it recognizes putative metabolites of NADH, such as N-methyl- and N-ribosyl-dihydronicotinamide. QR2 has been particularly associated with reactive oxygen species and memory, strongly suggesting a link among QR2 (as a possible key element in pro-oxidation), autophagy, and neurodegeneration. In molecular and cellular pharmacology, understanding physiopathological associations can be difficult because of a lack of specific and powerful tools. Here, we present a thorough description of the potent, nanomolar inhibitor [2-(2-methoxy-5H-1,4b,9-triaza(indeno[2,1-a]inden-10-yl)ethyl]-2-furamide (S29434 or NMDPEF; IC50 = 5-16 nM) of QR2 at different organizational levels. We provide full detailed syntheses, describe its cocrystallization with and behavior at QR2 on a millisecond timeline, show that it penetrates cell membranes and inhibits QR2-mediated reactive oxygen species (ROS) production within the 100 nM range, and describe its actions in several in vivo models and lack of actions in various ROS-producing systems. The inhibitor is fairly stable in vivo, penetrates cells, specifically inhibits QR2, and shows activities that suggest a key role for this enzyme in different pathologic conditions, including neurodegenerative diseases.

Concurrent Crystallization Mechanism Leading to Low Temperature Percolation of LAGP Glass-Ceramic Electrolyte.

Sintering of ceramic electrolytes (CE) is the most efficient way to obtain a dense, all ceramic solid-state battery with oxide-based materials. However, the high temperature required for this process leads to detrimental reactivity between CE and the active material. Crystalline ceramics are necessary for highly conductive oxide materials. Still, thermomechanical properties of glass-phase materials can be used to obtain a denser and more conductive CE. Glass-phase CE can be produced with Nasicon-type CE. Here, Li1.5Al0.5Ge1.5(PO4)3 (LAGP) glass is used as a model to investigate the formability, densification, and conduction properties upon crystallization. A complete study of the crystallization mechanism is first performed to fully understand how a high conductivity of 6.3 × 10-5 S·cm-1 at 30 °C with 92% relative density is obtained at a sintering temperature of only 550 °C without pressure. This is approximately 200 °C below the usual sintering temperature of LAGP. X-ray diffraction is then used to calculate the amount of crystalline phase as a function of time. A combined study of reaction kinetics and conductivity evolution reveals an autocatalytic nucleation effect, which produces an early crystallization pathway. Density is studied to quantify the ability of the glass to flow during the crystallization process.

NMR Study of Lithium Transport in Liquid-Ceramic Hybrid Solid Composite Electrolytes.

Despite their high conductivity, factors such as being fragile enough to face processing issues and interfacial incompatibility with lithium electrodes are some of the main drawbacks hindering the commercialization of inorganic (mainly oxide-based) solid electrolytes for use in all-solid-state lithium batteries. To this end, strategies such as the addition of solid polymer electrolytes have been proposed to improve the electrode-electrolyte interface. Hybrid electrolytes, which are usually composed of ceramic particles dispersed in a polymer, generally have a better affinity with electrodes and higher ionic conductivity than pure inorganic electrolytes. However, a significant downside of this strategy is that differences in lithium transportability between electrolyte layers can result in the formation of a high interfacial energy barrier across the cell. One strategy to ensure sufficient "wetting" of ceramics is to incorporate a liquid electrolyte directly into the solid inorganic electrolyte resulting in the formation of a hybrid liquid-ceramic electrolyte. To this end, liquid-ceramic hybrid electrolytes were prepared by adding LiG4TFSI, a solvate ionic liquid (SIL), to garnet, NASICON, and perovskite-type ceramic electrolytes. Although SIL addition resulted in increased ionic conductivity, comparisons between the pure SIL and the hybrid systems revealed that improvements were due to the SIL alone. A thorough investigation of the hybrid systems by solid-state nuclear magnetic resonance (NMR) revealed little to no lithium exchange between the ceramic and the SIL. This confirms that lithium conductivity preferentially occurs through the SIL in these hybrid systems. The primary role of the ceramic is to provide mechanical strength.

[Chemoinformatics and virtual screening of molecules for therapeutic use]. / Apports de la chémo-informatique dans la recherche et l'optimisation des molécules d'intérêt thérapeutique.

Successful identification of new chemical entities with drug-like properties in pharmaceutical and academic research groups involves an early screen and the use of a large number of public and proprietary chemical libraries. Before applying high-throughput experimental screening approaches, virtual screening strategies have been put in place in order to sort and filter this massive amount of compounds and data available at these very early stages. Chemoinformatic tools have a crucial role in this selection process and enable therapeutic chemists to focus very early on promising candidates. Virtual screening has conventionally been based either on models of the target or the ligand (molecule), but today these models include biopharmaceutical filters addressing right from the start of the project the ADMET (absorption, distribution, metabolism, excretion, and toxicity) properties of the molecules. Above all, chemoinformatic tools help chemists understand better the chemical diversity they can work with, especially when comparing chemical libraries. This paper will focus on exemples of the day-to-day use of chemoinformatics in screening programs. A large part will be dedicated to new tools (chemographic and pharmacographic approaches) being developed for the representation and analysis of chemical diversity, but also for combining chemical and biological information to expedite research programs.

S32826, a nanomolar inhibitor of autotaxin: discovery, synthesis and applications as a pharmacological tool.

Autotaxin catalyzes the transformation of lyso-phosphatidylcholine in lyso-phosphatidic acid (LPA). LPA is a phospholipid possessing a large panel of activity, in particular as a motility factor or as a growth signal, through its G-protein coupled seven transmembrane receptors. Indirect evidence strongly suggests that autotaxin is the main, if not the only source of circulating LPA. Because of its central role in pathologic conditions, such as oncology and diabetes/obesity, the biochemical properties of autotaxin has attracted a lot of attention, but confirmation of its role in pathology remains elusive. One way to validate and/or confirm its central role, is to find potent and selective inhibitors. A systematic screening of several thousand compounds using a colorimetric assay and taking advantage of the phosphodiesterase activity of autotaxin that requires the enzymatic site than for LPA generation, led to the discovery of a potent nanomolar inhibitor, [4-(tetradecanoylamino)benzyl]phosphonic acid (S32826). This compound was inhibitory toward the various autotaxin isoforms, using an assay measuring the [(14)C]lyso-phosphatidylcholine conversion into [(14)C]LPA. We also evaluated the activity of S32826 in cellular models of diabesity and oncology. Nevertheless, the poor in vivo stability and/or bioavailability of the compound did not permit to use it in animals. S32826 is the first reported inhibitor of autotaxin with an IC(50) in the nanomolar range that can be used to validate the role of autotaxin in various pathologies in cellular models.

Influence of foot position and vision on dynamic postural strategies during the "grand plié" ballet movement (squatting) in young and adult ballet dancers.

PURPOSE: To analyse dynamic postural strategies during the "grand plié" in two different foot positions (parallel or turned out), with and without vision, and as a function of age in ballet dancers. METHOD: Twenty young dancers (YD) aged from 8 to 16 years, and 20 adult dancers (AD) aged from 17 to 30 years were recruited. Center of pressure (CoP) and ground reaction forces (GRF) were recorded (500 Hz) during the grand plié (lowering, squatting and rising). This movement was tested with the feet parallel and with both lower limbs turned out (foot angle >140°), with eyes open (EO) and eyes closed (EC). Groups were compared using Student t-tests. Repeated analysis of variance was used to examine the effects of eyes and foot conditions, with a significance level of p < 0.05. RESULTS: The results of this study showed that dynamic postural strategies during the "grand plié" ballet movement are influenced by age, foot position and visual condition. CoP displacement length (p < 0.003) and CoP speed (p < 0.003) were higher in YD compared with AD. CoP surface (p < 0.05), mediolateral CoP speed (p < 0.048) and GRF parameters, particularly the mediolateral (p < 0.049), were higher than in the parallel than the turned out position. In both groups all CoP (p < 0.042) and GRF parameters (p < 0.049), except the vertical component, were higher with EC than EO. CONCLUSION: The effect of foot position was greatest with EO. The parallel position was less stable. The YD were more unstable in the parallel position, particularly with EC. For both groups, the lack of vision increased instability. These results show the importance of integrating balance training in a variety of foot positions and visual conditions, particularly during the initial stages of training to prevent injury.

The use of intraoperative ultrasound by anesthesiologists to facilitate the surgical management of peripheral nerve tumors of the upper extremity.

BACKGROUND: Traditional uses of ultrasound by anesthesiologists include transesophageal echocardiography, facilitation of vascular access, and guidance of peripheral nerve blocks. METHODS: In this case series, we report a novel application of ultrasound by anesthesiologists to facilitate the operative dissection of upper extremity peripheral nerve tumors. RESULTS: In four cases, ultrasound was used to intraoperatively locate the tumor, plan the safest surgical approach, and exclude tumor vascularity. CONCLUSIONS: Ultrasound can be used by anesthesiologists to facilitate the surgical management of peripheral nerve tumors.

Characterizing novice behavior associated with learning ultrasound-guided peripheral regional anesthesia.

BACKGROUND AND OBJECTIVES: Ultrasound-guided regional anesthesia is a rapidly growing field. There exists little information regarding the competencies involved with such a practice. The objective of this exploratory study was to characterize the behavior of novices as they undertook the challenges of learning a new technique. In addition to assessing for both committed errors and accuracy, we aimed to identify previously unrecognized quality-compromising behaviors that could help structure effective training interventions. METHODS: By using detailed video analyses, the performances of 6 anesthesia residents were evaluated while on a dedicated 1-month rotation in ultrasound-guided regional anesthesia. From these video reviews, we assessed accuracy, errors committed, performance times, and searched for previously unrecognized quality-compromising behaviors. RESULTS: A total of 520 nerve blocks were videotaped and reviewed. All residents performed at least 66 nerve blocks, with an overall success rate of 93.6% and 4 complications. Both speed and accuracy improved throughout the rotation. There were a total of 398 errors committed, with the 2 most common errors consisting of the failure to visualize the needle before advancement and unintentional probe movement. Five quality-compromising patterns of behavior were identified: (1) failure to recognize the maldistribution of local anesthesia, (2) failure to recognize an intramuscular location of the needle tip before injection, (3) fatigue, (4) failure to correctly correlate the sidedness of the patient with the sidedness of the ultrasound image, and (5) poor choice of needle-insertion site and angle with respect to the probe preventing accurate needle visualization. CONCLUSIONS: Based on the analysis of the committed errors and the identification of quality-compromising behaviors, we are able to recommend important targets for learning in future training and simulation programs.

Comparison of balance strategies in mountain climbers during real altitude exposure between 1.500m and 3.200m: Effects of age and expertise.

PURPOSE: The aim of this study was to estimate the effect of altitude (1500m vs. 3200m) on balance strategies, and to determine the influence of age (> or <40 years) and expertise (> or <20days of mountain climbing/year) on postural stability at altitude. METHOD: Eighty-nine volunteer mountaineers were tested at 1500m, 3200m immediately after exiting the cable car and at 3200m following the climb and return. Double-leg stance with eyes closed and single-leg stance with eyes open were tested. The center of pressure (CoP) was recorded for 12.8s using a stabilometric force platform. Length, surface, ratio (length/surface), and average speed of the CoP, as well as lower limb loading were calculated. Groups and altitudes were compared using ANOVA. Significance was set to p<0.05. RESULTS: CoP ratio was significantly higher during both balance conditions at 3200m compared to 1500m (p<0.05). Lower limb loading was symmetrical at 1500m but not at 3200m (p<0.05). There was no influence of age or expertise at 1500m (p=NS), however at 3200m, older subjects and subjects with less expertise had greater length and surface of the CoP, showing more instability (p<0.05). CONCLUSION: Postural stability was immediately reduced on arrival at 3200m, particularly in subjects above the age of 40 years with less expertise.

An abnormal clinical course of an ultrasound-guided supraclavicular brachial plexus block using 0.375% bupivacaine.

We report on the case of a reappearance of a supraclavicular nerve block after the apparent initiation of its resolution in a 21-year-old athlete undergoing repair of a valgus impaction syndrome of his right elbow. The patient's anesthetic management consisted of a supraclavicular nerve block and general anesthesia. The patient was discharged home with an apparent resolving nerve block. He returned to the hospital urgently when, at 7 hours after blockade, he lost all motor-sensory function in his arm. His workup ultimately yielded negative results, and the block resolved at 23 hours. In addition to documenting an abnormal course of a supraclavicular block, this case report questions the appropriateness of placing long-acting nerve blocks in outpatients.

Stereospecific synthesis of 5-substituted 2-bisarylthiocyclopentane carboxylic acids as specific matrix metalloproteinase inhibitors.

The synthesis and structure-activity relationship (SAR) studies of a series of cyclopentane carboxylic acid matrix metalloproteinase (MMP) inhibitors are described. Potent and specific MMP-2, -3, -9, -13 inhibitors were obtained by regio- and stereoselective substitutions at positions 2 and 5 on the cyclopentane ring. Compounds 2a and 2e are active in the mouse B16-F10 metastasis model and display very good pharmacokinetic parameters.

Improvement in the selectivity and metabolic stability of the serotonin 5-HT(1A) ligand, S 15535: a series of cis- and trans-2-(arylcycloalkylamine) 1-indanols.

S 15535 (1) displays a distinctive profile of agonist and antagonist (weak partial agonist) activity at pre- and postsynaptic 5-HT(1A) receptors, respectively. It has proven to be active in several models predictive of anxiolytic, antidepressant, and procognitive properties. In an attempt to increase its selectivity and metabolic stability, and guided by the results of human metabolic studies, we prepared a series of cis- and trans-2-(arylcycloalkylamine) 1-indanols. Irrespective of the nature of the arylcycloalkylamine moiety or the presence of substituents on the indanol ring, trans isomers invariably showed the highest affinity for human, recombinant h5-HT(1A) receptors. Among them, compounds 39, 42, 45, 49, 52, 53, 54, 57, 61, 64, 67, and 70 displayed similar or higher affinity than the parent compound 1 (pK(i) > or = 9.1). Lack of selectivity toward alpha1-adrenoceptors has been frequently encountered with 5-HT(1A) ligands. While S 15535 itself presents reasonable selectivity (158-fold) in this respect, trans piperazine derivatives 4-trans,35, 39, 41, 47, 64, 68, 69, 70, 71 displayed even more pronounced selectivity vs alpha1-adrenoceptors, with the nitro derivative 70 being highly selective (1259-fold). However, among the set of trans piperidines prepared, only 64, which also bears a nitro on the indanol ring, displayed selectivity greater than the parent compound 1. All trans derivatives behaved as partial agonists at h5-HT(1A) receptors, as determined by their submaximal stimulation of [35S]GTPgammaS binding to a level comparable to that observed with S 15535. In metabolic stability studies in vitro using human microsomes and hepatocytes, only trans piperazines and, in particular, 35, 39, 41, 68, 69, and 70, showed an improvement relative to 1, whereas trans piperidines did not. Compounds 35, 39, 41, and 70, which combined both improved selectivity and metabolic stability, and which retained the distinctive pharmacological characteristics of S 15535, were evaluated in animal models of anxiety. Of these, 35, which showed the highest oral bioavailability in vivo in rats, was resolved into its two isomers 36 and 37. The eutomer 37 displayed 47% oral bioavailability in the rat and was potently active (0.1-0.5 mg/kg, s.c.) in the rat ultrasonic vocalization and social interaction models, predictive of anxiolytic activity. In conclusion, 2-(arylcycloalkylamine) 1-indanols represent a novel class of potent 5-HT(1A) ligands in which the presence of the hydroxyl group in the benzylic position enhances selectivity, while substituents on the phenyl ring of the indanol moiety improve both selectivity and metabolic stability.

Synthesis and pharmacological evaluation of new 1,2-dithiolane based antioxidants.

Molecules containing a dithiolane moiety are widely investigated due to their antioxidant properties. The archetypal representative of this class of compounds is lipoic acid and indeed the lipoic acid-dihydrolipoic acid couple is part of the antioxidant defence system of the cell. In the course of a program aiming to find improved antioxidants effective in vivo, we designed, synthesised and pharmacologically investigated new lipoic acid analogs. The salient feature of these structures is the connection, via a thioamide or a thiocarbamate, of a 1,2-dithiolane moiety bearing a carbon chain and a N-alkyl-substituted morpholine ring. It was expected that the antioxidant and chelating properties of these functional groups combined with the basicity of the morpholine ring will impact on the antioxidant as well as on the partition and solubility characteristics of the compounds. Indeed in vitro and in vivo pharmacological investigation showed that these new molecules and especially those containing a thiocarbamate linker possess superior antioxidant properties compared with alpha-lipoic acid and to the amide or carbamate linker analogs. In particular, some of these compounds efficiently cross the blood brain barrier (BBB) thus providing efficient protection from lethality in a situation of induced oxidative stress. Moreover the absence of the 1,2-dithiolane moiety does not completely abolish antioxidant effects thus demonstrating that these compounds are distinct new chemical entities and not merely lipoic acid prodrugs. The chemical and pharmacological features of these new antioxidants are presented and discussed in the following paper.

QSAR Modelling of CYP3A4 Inhibition as a Screening Tool in the Context of DrugDrug Interaction Studies.

Drugdrug interaction potential (DDI), especially cytochrome P450 (CYP) 3A4 inhibition potential, is one of the most important parameters to be optimized before preclinical and clinical pharmaceutical development as regard to the number of marketed drug metabolized mainly by this CYP and potentially co-administered with the future drug. The present study aims to develop in silico models for CYP3A4 inhibition prediction to help medicinal chemists during the discovery phase and even before the synthesis of new chemical entities (NCEs), focusing on NCEs devoid of any inhibitory potential toward this CYP. In order to find a relevant relationship between CYP3A4 inhibition and chemical features of the screened compounds, we applied a genetic-algorithm-based QSAR exploratory tool SQS (Stochastic QSAR Sampler) in combination with different description approaches comprising alignment-independent Volsurf descriptors, ISIDA fragments and Topological Fuzzy Pharmacophore Triplets. The experimental data used to build models were extracted from an in-house database. We derived a model with good prediction ability that was confirmed on both newly synthesized compound and public dataset retrieved from Pubchem database. This model is a promising efficient tool for filtering out potentially problematic compounds.

Functional expression of sinusoidal drug transporters in primary human and rat hepatocytes.

Primary hepatocyte cultures are considered as a useful in vitro system for pharmacological/toxicological studies. Although expression of drug-metabolizing enzymes and canalicular drug transporters has been well documented in this cellular model, less information is available about sinusoidal drug transporter activities. This has led us to investigate functional expression of the major sinusoidal transporters in primary human and rat hepatocytes. Using radiolabeled substrates and chemical transporter inhibitors, activities of organic cation transporter 1, organic anion-transporting polypeptides, organic anion transporter 2, and Na(+)-taurocholate cotransporter were detected in cultured human and rat hepatocytes. In parallel, mRNA expression of these transporters was demonstrated using reverse transcriptase-quantitative polymerase chain reaction assays. Functional expression of sinusoidal transport proteins markedly decreased with time in primary rat hepatocyte cultures; by contrast, it remained relatively constant in primary human hepatocytes all along the culture, illustrating the fact that liver-specific functions, including drug-detoxifying pathways, are usually better preserved in cultured human hepatocytes than in their rodent counterparts. Primary hepatocytes, especially human hepatocytes, thus exhibit a pattern of sinusoidal transporter expression close to that found in vivo, highlighting the interest of hepatocyte cultures for drug detoxification studies.

Substituted benzocyloheptenes as potent and selective alpha(v) integrin antagonists.

A novel series of potent and specific alpha(v) integrin antagonists has been obtained by aminoalkyl substitutions on benzocyloheptene acetic acids as a rigid GD bioisostere. The preferred compounds 1-2, 1-3 and 1-8, showed nano- to subnanomolar IC(50) values on alpha(v)beta(3) and alpha(v)beta(5) integrins, with favorable pharmacokinetics.

Solid-phase synthesis of alpha-substituted 3-bisarylthio N-hydroxy propionamides as specific MMP inhibitors.

A novel series of potent and specific MMP-2,3,9,13 inhibitors has been obtained by modulation on solid phase by alpha and aryl substitutions on 3-arylthio-N-hydroxy-propionamides starting from itaconic acid.