Discovery of MK-8768, a Potent and Selective mGluR2 Negative Allosteric Modulator.

Glutamate plays a key role in cognition and mood, and it has been shown that inhibiting ionotropic glutamate receptors disrupts cognition, while enhancing ionotropic receptor activity is pro-cognitive. One approach to elevating glutamatergic tone has been to antagonize presynaptic metabotropic glutamate receptor 2 (mGluR2). A desire for selectivity over the largely homologous mGluR3 motivated a strategy to achieve selectivity through the identification of mGluR2 negative allosteric modulators (NAMs). Extensive screening and optimization efforts led to the identification of a novel series of 4-arylquinoline-2-carboxamides. This series was optimized for mGluR2 NAM potency, clean off-target activity, and desirable physical properties, which resulted in the identification of improved C4 and C7 substituents. The initial lead compound from this series was Ames-positive in a single strain with metabolic activation, indicating that a reactive metabolite was likely responsible for the genetic toxicity. Metabolic profiling and Ames assessment across multiple analogs identified key structure-activity relationships associated with Ames positivity. Further optimization led to the Ames-negative mGluR2 negative allosteric modulator MK-8768.

Parabolic achromatic color matching functions: Dependence on incremental and decremental luminance.

Either the brightness or lightness of a disk surrounded by an annulus is characterized in the most general case by a parabolic function of the annulus luminance when plotted on a log-log scale. This relationship has been modeled with a theory of achromatic color computation based on edge integration and contrast gain control [J. Vis.10, 1 (2010)1534-736210.1167/10.14.40]. We tested predictions of this model in new psychophysical experiments. Our results support the theory and reveal a previously unobserved property of parabolic matching functions that depends on the disk contrast polarity. We interpret this property in terms of a neural edge integration model incorporating data from macaque monkey physiology that indicates different physiological gain factors for incremental and decremental stimuli.

Circadian Rhythmicity of Vital Signs at Intensive Care Unit Discharge and Outcome of Traumatic Brain Injury.

BACKGROUND: Physiological functions with circadian rhythmicity are often disrupted during illness. OBJECTIVE: To assess the utility of circadian rhythmicity of vital signs in predicting outcome of traumatic brain injury (TBI). METHODS: A retrospective single-center cohort study of adult intensive care unit (ICU) patients with largely isolated TBI to explore the relationship between the circadian rhythmicity of vital signs during the last 24 hours before ICU discharge and clinical markers of TBI severity and score on the Glasgow Outcome Scale 6 months after injury (GOS-6). RESULTS: The 130 study participants had a median age of 39.0 years (IQR, 23.0-59.0 years), a median Glasgow Coma Scale score at the scene of 8.0 (IQR, 3.0-13.0), and a median Rotterdam score on computed tomography of the head of 3 (IQR, 3-3), with 105 patients (80.8%) surviving to hospital discharge. Rhythmicity was present for heart rate (30.8% of patients), systolic blood pressure (26.2%), diastolic blood pressure (20.0%), and body temperature (26.9%). Independent predictors of a dichotomized GOS-6 ≥4 were the Rotterdam score (odds ratio [OR], 0.38 [95% CI, 0.18-0.81]; P = .01), Glasgow Coma Scale score at the scene (OR, 1.22 [95% CI, 1.05-1.41]; P = .008), age (OR, 0.95 [95% CI, 0.92-0.98]; P = .003), oxygen saturation <90% in the first 24 hours (OR, 0.19 [95% CI, 0.05-0.73]; P = .02), serum sodium level <130 mmol/L (OR, 0.20 [95% CI, 0.05-0.70]; P = .01), and active intracranial pressure management (OR, 0.16 [95% CI, 0.04-0.62]; P = .008), but not rhythmicity of any vital sign. CONCLUSION: Circadian rhythmicity of vital signs at ICU discharge is not predictive of GOS-6 in patients with TBI.

The PET tracer [11C]MK-6884 quantifies M4 muscarinic receptor in rhesus monkeys and patients with Alzheimer's disease.

Positron emission tomography (PET) ligands play an important role in the development of therapeutics by serving as target engagement or pharmacodynamic biomarkers. Here, we describe the discovery and translation of the PET tracer [11C]MK-6884 from rhesus monkeys to patients with Alzheimer's disease (AD). [3H]MK-6884/[11C]MK-6884 binds with high binding affinity and good selectivity to an allosteric site on M4 muscarinic cholinergic receptors (M4Rs) in vitro and shows a regional distribution in the brain consistent with M4R localization in vivo. The tracer demonstrates target engagement of positive allosteric modulators of the M4R (M4 PAMs) through competitive binding interactions. [11C]MK-6884 binding is enhanced in vitro by the orthosteric M4R agonist carbachol and indirectly in vivo by the acetylcholinesterase inhibitor donepezil in rhesus monkeys and healthy volunteers, consistent with its pharmacology as a highly cooperative M4 PAM. PET imaging of [11C]MK-6884 in patients with AD identified substantial regional differences quantified as nondisplaceable binding potential (BPND) of [11C]MK-6884. These results suggest that [11C]MK-6884 is a useful target engagement biomarker for M4 PAMs but may also act as a sensitive probe of neuropathological changes in the brains of patients with AD.

Duration Comparisons for Vision and Touch Are Dependent on Presentation Order and Temporal Context.

Integrating visual and tactile information in the temporal domain is critical for active perception. To accomplish this, coordinated timing is required. Here, we study perceived duration within and across these two modalities. Specifically, we examined how duration comparisons within and across vision and touch were influenced by temporal context and presentation order using a two-interval forced choice task. We asked participants to compare the duration of two temporal intervals defined by tactile or visual events. Two constant standard durations (700 ms and 1,000 ms in 'shorter' sessions; 1,000 ms and 1,500 ms in 'longer' sessions) were compared to variable comparison durations in different sessions. In crossmodal trials, standard and comparison durations were presented in different modalities, whereas in the intramodal trials, the two durations were presented in the same modality. The standard duration was either presented first () or followed the comparison duration (). In both crossmodal and intramodal conditions, we found that the longer standard duration was overestimated in trials and underestimated in trials whereas the estimation of shorter standard duration was unbiased. Importantly, the estimation of 1,000ms was biased when it was the longer standard duration within the shorter sessions but not when it was the shorter standard duration within the longer sessions, indicating an effect of temporal context. The effects of presentation order can be explained by a central tendency effect applied in different ways to different presentation orders. Both crossmodal and intramodal conditions showed better discrimination performance for trials than trials, supporting the Type B effect for both crossmodal and intramodal duration comparison. Moreover, these results were not dependent on whether the standard duration was defined using tactile or visual stimuli. Overall, our results indicate that duration comparison between vision and touch is dependent on presentation order and temporal context, but not modality.

Structures of active-state orexin receptor 2 rationalize peptide and small-molecule agonist recognition and receptor activation.

Narcolepsy type 1 (NT1) is a chronic neurological disorder that impairs the brain's ability to control sleep-wake cycles. Current therapies are limited to the management of symptoms with modest effectiveness and substantial adverse effects. Agonists of the orexin receptor 2 (OX2R) have shown promise as novel therapeutics that directly target the pathophysiology of the disease. However, identification of drug-like OX2R agonists has proven difficult. Here we report cryo-electron microscopy structures of active-state OX2R bound to an endogenous peptide agonist and a small-molecule agonist. The extended carboxy-terminal segment of the peptide reaches into the core of OX2R to stabilize an active conformation, while the small-molecule agonist binds deep inside the orthosteric pocket, making similar key interactions. Comparison with antagonist-bound OX2R suggests a molecular mechanism that rationalizes both receptor activation and inhibition. Our results enable structure-based discovery of therapeutic orexin agonists for the treatment of NT1 and other hypersomnia disorders.

Exploring the Determinants of Color Perception Using #Thedress and Its Variants: The Role of Spatio-Chromatic Context, Chromatic Illumination, and Material-Light Interaction.

The colors that people see depend not only on the surface properties of objects but also on how these properties interact with light as well as on how light reflected from objects interacts with an individual's visual system. Because individual visual systems vary, the same visual stimulus may elicit different perceptions from different individuals. #thedress phenomenon drove home this point: different individuals viewed the same image and reported it to be widely different colors: blue and black versus white and gold. This phenomenon inspired a collection of demonstrations presented at the Vision Sciences Society 2015 Meeting which showed how spatial and temporal manipulations of light spectra affect people's perceptions of material colors and illustrated the variability in individual color perception. The demonstrations also explored the effects of temporal alterations in metameric lights, including Maxwell's Spot, an entoptic phenomenon. Crucially, the demonstrations established that #thedress phenomenon occurs not only for images of the dress but also for the real dress under real light sources of different spectral composition and spatial configurations.

The Pre-Hospital Initial Fluid Therapy Estimate in Early Nasty Burns (PHIFTEEN B, 15-B) Guideline applied to a retrospective cohort of Intensive Care Unit patients with major burns.

BACKGROUND: Appropriate fluid administration in severe burns is a cornerstone of early burns management. The American Burns Association's (ABA) recommendation is to administer 2 mL-4 mL × burnt Body Surface Area (BSA) × weight in the first 24 h with half administered in the first eight hours. Unfortunately, the calculations involved are complex and clinicians do not estimate the BSA or weight well, which can lead to errors in the amount of fluid administered. To simplify cognitive load to calculate the fluid resuscitation of early burns, the investigators derived the PHIFTEEN B (15-B) guideline. The 15-B guideline estimates the initial hourly fluid for adults ≥ 50 kg to be: 15 mL × BSA (to the nearest 10%) AIMS: To model and determine the accuracy of the 15-B calculated based on the characteristics of a retrospective cohort of patients admitted with ≥ 20% BSA to the Royal Brisbane and Women's Hospital (RBWH) Intensive Care Unit (ICU). METHODS: The 15-B formula was retrospectively calculated on the prehospital BSA estimate on patients admitted to the RBWH ICU. In addition, the 15-B guideline was modelled against a variety of weights and BSAs. The fluid volume was deemed to be clinically significant if it was greater than 250 mL/h outside the ABA's recommendations. RESULTS: The ICU cohort consisted of 107 patients (63.2% male, median age 37 years), with a median ICU estimated BSA of 40% and a median ICU weight estimation of 80 kg. In 43.9% of the cohort, the magnitude of the proportional difference between prehospital and ICU BSA estimate was greater than 25%. The 15-B formula accurately estimated the hourly fluid for all BSA (20%-100%) and weight combinations (50 kg-140 kg) in a BSA- weight matrix. When prehospital BSA estimate was utilized, 15-B guideline accurately estimated the fluid to be given within clinically significant limits for 97.2% of cases. CONCLUSIONS: The 15-B formula is a simple, easy to calculate guideline which approximates the early fluid estimates in severely burned patients despite inaccuracy in prehospital BSA estimates.

Discovery of [11C]MK-6884: A Positron Emission Tomography (PET) Imaging Agent for the Study of M4Muscarinic Receptor Positive Allosteric Modulators (PAMs) in Neurodegenerative Diseases.

The measurement of receptor occupancy (RO) using positron emission tomography (PET) has been instrumental in guiding discovery and development of CNS directed therapeutics. We and others have investigated muscarinic acetylcholine receptor 4 (M4) positive allosteric modulators (PAMs) for the treatment of symptoms associated with neuropsychiatric disorders. In this article, we describe the synthesis, in vitro, and in vivo characterization of a series of central pyridine-related M4 PAMs that can be conveniently radiolabeled with carbon-11 as PET tracers for the in vivo imaging of an allosteric binding site of the M4 receptor. We first demonstrated its feasibility by mapping the receptor distribution in mouse brain and confirming that a lead molecule 1 binds selectively to the receptor only in the presence of the orthosteric agonist carbachol. Through a competitive binding affinity assay and a number of physiochemical properties filters, several related compounds were identified as candidates for in vivo evaluation. These candidates were then radiolabeled with 11C and studied in vivo in rhesus monkeys. This research eventually led to the discovery of the clinical radiotracer candidate [11C]MK-6884.

Neurocomputational Lightness Model Explains the Appearance of Real Surfaces Viewed Under Gelb Illumination.

One of the primary functions of visual perception is to represent, estimate, and evaluate the properties of material surfaces in the visual environment. One such property is surface color, which can convey important information about ecologically relevant object characteristics such as the ripeness of fruit and the emotional reactions of humans in social interactions. This paper further develops and applies a neural model (Rudd, 2013, 2017) of how the human visual system represents the light/dark dimension of color-known as lightness-and computes the colors of achromatic material surfaces in real-world spatial contexts. Quantitative lightness judgments conducted with real surfaces viewed under Gelb (i.e., spotlight) illumination are analyzed and simulated using the model. According to the model, luminance ratios form the inputs to ON- and OFF-cells, which encode local luminance increments and decrements, respectively. The response properties of these cells are here characterized by physiologically motivated equations in which different parameters are assumed for the two cell types. Under non-saturating conditions, ON-cells respond in proportion to a compressive power law of the local incremental luminance in the image that causes them to respond, while OFF-cells respond linearly to local decremental luminance. ON- and OFF-cell responses to edges are log-transformed at a later stage of neural processing and then integrated across space to compute lightness via an edge integration process that can be viewed as a neurally elaborated version of Land's retinex model (Land & McCann, 1971). It follows from the model assumptions that the perceptual weights-interpreted as neural gain factors-that the model observer applies to steps in log luminance at edges in the edge integration process are determined by the product of a polarity-dependent factor 1-by which incremental steps in log luminance (i.e., edges) are weighted by the value <1.0 and decremental steps are weighted by 1.0-and a distance-dependent factor 2, whose edge weightings are estimated to fit perceptual data. The model accounts quantitatively (to within experimental error) for the following: lightness constancy failures observed when the illumination level on a simultaneous contrast display is changed (Zavagno, Daneyko, & Liu, 2018); the degree of dynamic range compression in the staircase-Gelb paradigm (Cataliotti & Gilchrist, 1995; Zavagno, Annan, & Caputo, 2004); partial releases from compression that occur when the staircase-Gelb papers are reordered (Zavagno, Annan, & Caputo, 2004); and the larger compression release that occurs when the display is surrounded by a white border (Gilchrist & Cataliotti, 1994).

Structural basis for selectivity and diversity in angiotensin II receptors.

The angiotensin II receptors AT1R and AT2R serve as key components of the renin-angiotensin-aldosterone system. AT1R has a central role in the regulation of blood pressure, but the function of AT2R is unclear and it has a variety of reported effects. To identify the mechanisms that underlie the differences in function and ligand selectivity between these receptors, here we report crystal structures of human AT2R bound to an AT2R-selective ligand and to an AT1R/AT2R dual ligand, capturing the receptor in an active-like conformation. Unexpectedly, helix VIII was found in a non-canonical position, stabilizing the active-like state, but at the same time preventing the recruitment of G proteins or ß-arrestins, in agreement with the lack of signalling responses in standard cellular assays. Structure-activity relationship, docking and mutagenesis studies revealed the crucial interactions for ligand binding and selectivity. Our results thus provide insights into the structural basis of the distinct functions of the angiotensin receptors, and may guide the design of new selective ligands.

Brightness in human rod vision depends on slow neural adaptation to quantum statistics of light.

In human rod-mediated vision, threshold for small, brief flashes rises in proportion to the square root of adapting luminance at all but the lowest and highest adapting intensities. A classical signal detection theory from Rose (1942, 1948) and de Vries (1943) attributed this rise to the perceptual masking of weak flashes by Poisson fluctuations in photon absorptions from the adapting field. However, previous work by Brown and Rudd (1998) demonstrated that the square-root law also holds for suprathreshold brightness judgments, a finding that supports an alternative explanation of the square-root sensitivity changes as a consequence of physiological adaptation (i.e., neural gain control). Here, we employ a dichoptic matching technique to investigate the properties of this brightness gain control. We show that the brightness gain control: 1) affects the brightness of high-intensity suprathreshold flashes for which assumptions of the de Vries-Rose theory are strongly violated; 2) exhibits a long time course of 100-200 s; and 3) is subject to modulation by temporal contrast noise when the mean adapting luminance is held constant. These findings are consistent with the hypothesis that the square-root law results from a slow neural adaptation to statistical noise in the rod pool. We suggest that such adaptation may function to reduce the probability of spurious ganglion cell spiking activity due to photon fluctuation noise as the ambient illumination level is increased.

Hepatitis C virus NS3/4a protease inhibitors.

Hepatitis C virus (HCV) infection is a major health issue around the world and HCV NS3/4a protease inhibitors have been the focus of intensive research for the past 20 years. From the first identification of substrate-derived peptide inhibitors to the complex, macrocyclic compounds, including paritaprevir and grazoprevir, that are currently available, the field has used structure-based design to confront the issues of potency, resistance and pharmacokinetics. Numerous breakthrough structures from a multitude of companies have led to compounds that are now key components of combination therapies with cure rates of >90%. Herein, we detail the compounds that have advanced to clinical trials including their design and their impact on the NS3/4a protease field.

Interim pressure garment therapy (4-6 mmHg) and its effect on donor site healing in burn patients: study protocol for a randomised controlled trial.

BACKGROUND: Pressure garment therapy (PGT) is well accepted and commonly used by clinicians in the treatment of burns scars and grafts. The medium to high pressures (24-40 mmHg) in these garments can support scar minimisation, and evidence is well documented for this particular application. However, PGT specifically for burn donor sites, of which a sequela is also scarring, is not well documented. This study protocol investigates the impact of a low pressure (4-6 mmHg) interim garment on donor site healing and scarring. With a primary purpose of holding donor dressings in place, the application of the interim pressure garment (IPG) appears to have been twofold. IPGs for donor sites have involved inconsistent application with a focus on securing wound dressing rather than scar management. However, anecdotal and observational evidence suggests that IPGs also make a difference to some patient's scar outcomes for donor sites. This study protocol outlines a randomised controlled trial designed to test the effectiveness of this treatment on reducing scarring to burn donor sites. METHODS/DESIGN: This study is a single-centre, single (assessor)-blinded, randomised control trial in patients with burns donor sites to their thighs. Patients will be randomly allocated to a control group (with no compression to donor sites) or to an experimental group (with compression to donor sites) as the comparative treatment. Groups will be compared at baseline regarding the important prognostic indicators: donor site location, depth, size, age, and time since graft (5 days). The IPG treatment will be administered post-operatively (on day 5). Follow-up assessments and garment replacement will be undertaken fortnightly for a period of 2 months. DISCUSSION: This study focuses on a unique area of burns scar management using a low-pressure tubular support garment for the reduction of donor site scars. Such therapy specifically for donor scar management is poorly represented in the literature. This study was designed to test a potentially cost-effective scar prevention for patients with donor sites to the thigh. No known studies of this nature have been carried out to date, and there is a need for rigorous clinical evidence for low-pressure support garments for donor site scar minimisation. TRIAL REGISTRATION: Australian New Zealand Clinical Trials Registry identifier ACTRN12610000127000 . Registered 8 Mar 2010.

Identification and in Vivo Evaluation of Liver X Receptor ß-Selective Agonists for the Potential Treatment of Alzheimer's Disease.

Herein, we describe the development of a functionally selective liver X receptor ß (LXRß) agonist series optimized for Emax selectivity, solubility, and physical properties to allow efficacy and safety studies in vivo. Compound 9 showed central pharmacodynamic effects in rodent models, evidenced by statistically significant increases in apolipoprotein E (apoE) and ATP-binding cassette transporter levels in the brain, along with a greatly improved peripheral lipid safety profile when compared to those of full dual agonists. These findings were replicated by subchronic dosing studies in non-human primates, where cerebrospinal fluid levels of apoE and amyloid-ß peptides were increased concomitantly with an improved peripheral lipid profile relative to that of nonselective compounds. These results suggest that optimization of LXR agonists for Emax selectivity may have the potential to circumvent the adverse lipid-related effects of hepatic LXR activity.

The phantom illusion.

It is well known that visible luminance gradients may generate contrast effects. In this work we present a new paradoxical illusion in which the luminance range of gradual transitions has been reduced to make them invisible. By adopting the phenomenological method proposed by Kanizsa, we have found that unnoticeable luminance gradients still generate contrast effects. But, most interestingly, we have found that when their width is narrowed, rather than generating contrast effects on the surrounded surfaces, they generate an assimilation effect. Both high- and low-level interpretations of this "phantom" illusion are critically evaluated.

P2-quinazolinones and bis-macrocycles as new templates for next-generation hepatitis C virus NS3/4a protease inhibitors: discovery of MK-2748 and MK-6325.

With the goal of identifying inhibitors of hepatitis C virus (HCV) NS3/4a protease that are potent against a wide range of genotypes and clinically relevant mutant viruses, several subseries of macrocycles were investigated based on observations made during the discovery of MK-5172. Quinazolinone-containing macrocycles were identified as promising leads, and optimization for superior cross-genotype and mutant enzyme potency as well as rat liver and plasma concentrations following oral dosing, led to the development of MK-2748. Additional investigation of a series of bis-macrocycles containing a fused 18- and 15-membered ring system were also optimized for the same properties, leading to the discovery of MK-6325. Both compounds display the broad genotype and mutant potency necessary for clinical development as next-generation HCV NS3/4a protease inhibitors.

High-Throughput Screen of GluK1 Receptor Identifies Selective Inhibitors with a Variety of Kinetic Profiles Using Fluorescence and Electrophysiology Assays.

GluK1, a kainate subtype of ionotropic glutamate receptors, exhibits an expression pattern in the CNS consistent with involvement in pain processing and migraine. Antagonists of GluK1 have been shown to reduce pain signaling in the spinal cord and trigeminal nerve, and are predicted to provide pain and migraine relief. We developed an ultra-high-throughput small-molecule screen to identify antagonists of GluK1. Using the calcium indicator dye fluo-4, a multimillion-member small-molecule library was screened in 1536-well plate format on the FLIPR (Fluorescent Imaging Plate Reader) Tetra against cells expressing a calcium-permeable GluK1. Following confirmation in the primary assay and subsequent counter-screen against the endogenous Par-1 receptor, 6100 compounds were selected for dose titration to assess potency and selectivity. Final triage of 1000 compounds demonstrating dose-dependent inhibition with IC50 values of less than 12 µM was performed in an automated whole-cell patch clamp electrophysiology assay. Although a weak correlation between electrophysiologically active and calcium-imaging active compounds was observed, the identification of electrophysiologically active compounds with a range of kinetic profiles revealed a broad spectrum of mechanisms of action.

Acute vs. chronic stressors, multiple suicide attempts, and persistent suicide ideation in US soldiers.

This study examined recent-onset (i.e., acute) and persistent (i.e., chronic) life stressors among 54 acutely suicidal US Army Soldiers and examined their relationship to persistence of suicidal crises over time. Soldiers with a history of multiple suicide attempts reported the most severe suicide ideation (F(2,51) = 4.18, p = 0.021) and the greatest number of chronic stressors (F(2,51) = 5.11, p = 0.009). Chronic but not acute stressors were correlated with severity of suicide ideation (r = 0.24, p = 0.026). Participants reporting low-to-average levels of chronic stress resolved suicide ideation during the 6-month follow-up, but participants reporting high levels of chronic stress did not (Wald χ(1) = 4.57, p = 0.032). Soldiers who are multiple attempters report a greater number of chronic stressors. Chronic, but not acute-onset, stressors are associated with more severe and longer-lasting suicidal crises.

A cortical edge-integration model of object-based lightness computation that explains effects of spatial context and individual differences.

Previous work has demonstrated that perceived surface reflectance (lightness) can be modeled in simple contexts in a quantitatively exact way by assuming that the visual system first extracts information about local, directed steps in log luminance, then spatially integrates these steps along paths through the image to compute lightness (Rudd and Zemach, 2004, 2005, 2007). This method of computing lightness is called edge integration. Recent evidence (Rudd, 2013) suggests that human vision employs a default strategy to integrate luminance steps only along paths from a common background region to the targets whose lightness is computed. This implies a role for gestalt grouping in edge-based lightness computation. Rudd (2010) further showed the perceptual weights applied to edges in lightness computation can be influenced by the observer's interpretation of luminance steps as resulting from either spatial variation in surface reflectance or illumination. This implies a role for top-down factors in any edge-based model of lightness (Rudd and Zemach, 2005). Here, I show how the separate influences of grouping and attention on lightness can be modeled in tandem by a cortical mechanism that first employs top-down signals to spatially select regions of interest for lightness computation. An object-based network computation, involving neurons that code for border-ownership, then automatically sets the neural gains applied to edge signals surviving the earlier spatial selection stage. Only the borders that survive both processing stages are spatially integrated to compute lightness. The model assumptions are consistent with those of the cortical lightness model presented earlier by Rudd (2010, 2013), and with neurophysiological data indicating extraction of local edge information in V1, network computations to establish figure-ground relations and border ownership in V2, and edge integration to encode lightness and darkness signals in V4.