Search

Objective. Transforming the University of Southern California health care system requires that institutions and organizations position equity, diversity, inclusion (EDI), and anti-racism central to their missions. The purpose of this administrative case report was to describe a systematic approach taken by an academic physical therapy department to develop a comprehensive antiracism plan that engages all interested and affected parties and includes processes for sustainable, long-term engagement. Methods. Four strategies contributed to organizational change towards anti-racism: Holding Ourselves Accountable; Developing a Plan; Building Consensus; and Providing Education, Support, and Resources. The attitudes of faculty and staff about racism and anti-racist actions were assessed through surveys at the start of the process and after and 1 year. Engagement in activities, meetings, and trainings related to EDI and anti-racism were logged for faculty and staff. Results. From November 2020 through November 2021, several outcomes were achieved including: making structural organizational changes; including EDI in the faculty merit review; developing a bias reporting mechanism; establishing faculty development activities, resources, and groups; and implementing structured efforts to recruit a diverse cohort. Within that year, faculty and staff engaged in 99.32 hours of EDI and anti-racism trainings, workshops, and resource groups. Survey data showed persistent high support and commitment to EDI and anti-racism. Faculty and staff reported that they felt more equipped to identify and address individual and institutional racism and they reported risking their reputations to talk about race more often. Confidence in their ability to identify and resolve conflicts related to microaggressions, cultural insensitivity, and bias improved. However, their self-reported ability to identify and address structural racism remained unchanged. Conclusion. By approaching anti-racism as transformative rather than performative, an academic physical therapy department was able to develop and implement a comprehensive anti-racism plan with high support and engagement. Impact. The physical therapy profession has not been immune to racism and health injustice. Organizational change to become anti-racist is imperative for excellence and a necessary challenge to undertake if the physical therapy profession is to transform society and improve the human experience.

Purpose. Maintaining balance during static standing postures requires the coordination of many neuromuscular mechanisms. The role of the intrinsic and extrinsic foot muscles in this paradigm has yet to be clearly defined. The purpose of this study was to explore foot muscle activation during static phases on common weight-bearing tasks of varying loads and balance demands. Methods. Twenty healthy young adults performed 6 standing postures (single-limb and double-limb stand, squat, and heel raise) with one foot on a force plate. Muscle activity was recorded from the abductor hallucis, flexor hallucis longus and brevis, and tibialis posterior using intramuscular electrodes; surface electrodes were used to record activity from the peroneus longus and tibialis anterior. Two-way repeated measures ANOVA (2 loading conditions × 3 postures) were run to compare muscle activation and center of pressure velocity. Results. Intrinsic foot muscle activity increased as loading and postural demand increased; however, the specific effects varied for each of the extrinsic foot muscles. Conclusions. These results suggest that the intrinsic foot muscles play an important role in maintaining static balance. Strengthening intrinsic and extrinsic foot muscles may help increase stability in people who have weak toe flexors or who suffer from a variety of foot pathologies.

Purpose: It is not well established how motion and muscle activation of the medial longitudinal arch (MLA) of the foot vary under different loading conditions. Intrinsic and extrinsic foot muscles may play a role in postural control, which may be investigated by comparing loading tasks with differing postural demands. The objective of this study was to investigate the interaction of MLA flexibility and loading task on muscle activation. Methods: Twenty healthy adults completed two instrumented single-foot loading tasks: controlled external load of 50% body weight while sitting and bilateral standing. Fine-wire intramuscular and surface electromyography collected flexor hallucis brevis, abductor hallucis, tibialis posterior, flexor hallucis longus, tibialis anterior, and peroneus longus activation. MLA deformation was measured as a percent change in navicular height with loading. Results: During seated external loading, greater MLA deformation was associated with greater muscle activation for all instrumented muscles (R2 = 0.224–0.303, p < 0.05) except for tibialis anterior. During bilateral stance, there were no correlations between MLA deformation and muscle activation. Activation of all extrinsic muscles except for tibialis anterior were greater during bilateral standing than during external loading (p = 0.002–0.013), indicating activation of these muscles was caused by postural demands of the standing task, not simply load. Conclusions: MLA deformation and muscle activation are strongly task-dependent.

Benzylisoquinoline alkaloids (BIAs) are a structurally diverse class of plant-specialized metabolites that have been particularly well-studied in the order Ranunculales. The N-methyltransferases (NMTs) in BIA biosynthesis can be divided into three groups according to substrate specificity and amino acid sequence. Here, we report the first crystal structures of enzyme complexes from the tetrahydroprotoberberine NMT (TNMT) subclass, specifically for GfTNMT from the yellow horned poppy (Glaucium flavum). GfTNMT was co-crystallized with the cofactor S-adenosyl-l-methionine (dmin = 1.6 A), the product S-adenosyl-l-homocysteine (dmin = 1.8 A), or in complex with S-adenosyl-l-homocysteine and (S)-cis-N-methylstylopine (dmin = 1.8 A). These structures reveal for the first time how a mostly hydrophobic L-shaped substrate recognition pocket selects for the (S)-cis configuration of the two central six-membered rings in protoberberine BIA compounds. Mutagenesis studies confirm and functionally define the roles of several highly-conserved residues within and near the GfTNMT-active site. The substrate specificity of TNMT enzymes appears to arise from the arrangement of subgroup-specific stereospecific recognition elements relative to catalytic elements that are more widely-conserved among all BIA NMTs. The binding mode of protoberberine compounds to GfTNMT appears to be similar to coclaurine NMT, with the isoquinoline rings buried deepest in the binding pocket. This binding mode differs from that of pavine NMT, in which the benzyl ring is bound more deeply than the isoquinoline rings. The insights into substrate recognition and catalysis provided here form a sound basis for the rational engineering of NMT enzymes for chemoenzymatic synthesis and metabolic engineering.

The purpose of the study was to characterize the Balance-Dexterity Task as a means to investigate a concurrent bipedal lower-extremity task and trunk control during dynamic balance. The task combines aspects of single-limb balance and the lower-extremity dexterity test by asking participants to stand on one limb while compressing an unstable spring with the contralateral limb to an individualized target force. Nineteen non-disabled participants completed the study, and performance measures for the demands of each limb – balance and dexterous force control – as well as kinematic and electromyographic measures of trunk control were collected. Given five practice trials, participants achieved compression forces ranging from 100 to 139 N (mean 121.2 ± 12.3 N), representing 14.4–23.0% of body weight (mean 18.7 ± 2.4%), which were then presented as target forces during test trials. Dexterous force control coefficient of variation and average magnitude of the center of pressure (COP) resultant velocity were associated such that greater variability in force control was accompanied by greater COP velocity (R = 0.598, p = 0.007). Trunk coupling, quantified as the coefficient of determination (R2) of a frontal plane thorax and pelvis angle-angle plot, varied independently of any measure of balance or dexterous force control. The Balance-Dexterity Task is a continuous, dynamic balance task where bipedal coordination and trunk coupling can be concurrently observed and studied.

OBJECTIVE: Altered ground reaction force (GRF) and joint torsional stiffness are associated with various lower extremity injuries, but these have yet to be examined in dancers with flexor hallucis longus (FHL) tendinopathy. Additionally, a simple, field-friendly kinematic correlate to ground contact kinetics would be useful for clinical application. The purpose of this study was to compare lower extremity biomechanics during takeoff of a dance leap (saut de chat) in dancers with and without FHL tendinopathy, and to examine lower limb posture at initial contact as a clinical correlate of injury-related kinetic factors. METHODS: Motion capture and inverse dynamics were used to analyze saut de chat takeoff performed by 11 uninjured dancers and 8 dancers with FHL tendinopathy. GRF parameters, joint torsional stiffness of the metatarsophalangeal, ankle, and knee joints, and lower extremity posture at initial contact were compared between groups using Welch’s t-tests. RESULTS: Dancers with FHL tendinopathy maintained similar jump height as the uninjured dancers, but exhibited lower peak vertical GRF, longer time to peak force, and less joint torsional stiffness at the metatarsophalangeal, ankle, and knee joints during loading response of the takeoff step. Lower extremity contact angle was smaller and the horizontal distance between center-of-mass and center-of-pressure was greater in dancers with FHL tendinopathy. These two measures of lower limb posture at initial contact were significantly correlated with kinetic factors occurring later in ground contact (R2=0.29-0.51). CONCLUSION: Dancers with FHL tendinopathy demonstrated altered lower extremity kinetics during takeoff of a leap compared to uninjured dancers, which may contribute to, or be a compensation response to, injury

This study investigated effects of cognitive dual-task interference and task prioritization instructions on task performance and trunk control during a dynamic balance task in persons with and without recurrent low back pain (rLBP). First, we tested the hypothesis that those with rLBP rely more on cognitive resources than back-healthy controls, and therefore trunk kinematics would be altered under dual-task interference conditions. Then, we tested participants’ ability to modulate task performance in accord with prioritization instructions. Persons with and without rLBP (n = 19/group) performed the Balance-Dexterity Task, which involved single-limb balance while compressing an unstable spring with the other limb, with and without a cognitive task engaging verbal working memory. Trunk coupling was quantified with the coefficient of determination (R2) of an angle–angle plot of thorax–pelvis frontal plane motion. Task performance was quantified using variability of spring compression force and of cognitive task errors. Trunk coupling in the rLBP group was lower than that of the back-healthy control group in the single-task condition (p = 0.024) and increased in the dual-task condition (p = 0.006), abolishing the difference between groups. Significant main effects of task prioritization instruction on performance were observed with no differences between groups, indicating similar performance modulation. Cognitive task error variability decreased with a switch from a single- to dual-task condition, exposing an unexpected facilitation effect. We interpret these findings in the context of movement-specific reinvestment and action-specific perception theories as they pertain to cognitive contributions to posture and how the dual-task interference paradigm may influence those contributions.

Coordination of the trunk and hips is crucial for successful dynamic balance in many activities of daily living. Persons with recurrent low back pain (rLBP), both while symptomatic and during periods of symptom remission, exhibit dysfunctional muscle activation patterns and coordination of these joints. In a novel dynamic balance task where persons in remission from rLBP exhibit dissociated trunk motion, it is unknown how trunk and hip musculature are coordinated. Activation of hip and trunk muscles were acquired from nineteen persons with and without rLBP during the Balance-Dexterity Task, which involves balancing on one limb while compressing an unstable spring with the other. There were no between-group differences in activation amplitude for any muscle groups tested. In back-healthy control participants, hip and trunk muscle activation amplitudes increased proportionally in response to the added instability of the spring (R = 0.837, p < 0.001). Increases in muscle activation amplitudes in the group in remission from rLBP were not proportional (R = 0.113, p = 0.655). Instead, hip muscle activation in this group was associated with task performance, i.e. dexterous control of the spring (R = 0.676, p = 0.002). These findings highlight atypical coordination of hip and trunk musculature potentially related to task demands in persons with rLBP even during remission from pain.

Benzylisoquinoline alkaloids (BIAs) are a structurally diverse class of plant-specialized metabolites that have been particularly well-studied in the order Ranunculales. The N-methyltransferases (NMTs) in BIA biosynthesis can be divided into three groups according to substrate specificity and amino acid sequence. Here, we report the first crystal structures of enzyme complexes from the tetrahydroprotoberberine NMT (TNMT) subclass, specifically for GfTNMT from the yellow horned poppy (Glaucium flavum). GfTNMT was co-crystallized with the cofactor S-adenosyl-l-methionine (d(min) = 1.6 Å), the product S-adenosyl-l-homocysteine (d(min) = 1.8 Å), or in complex with S-adenosyl-l-homocysteine and (S)-cis-N-methylstylopine (d(min) = 1.8 Å). These structures reveal for the first time how a mostly hydrophobic L-shaped substrate recognition pocket selects for the (S)-cis configuration of the two central six-membered rings in protoberberine BIA compounds. Mutagenesis studies confirm and functionally define the roles of several highly-conserved residues within and near the GfTNMT-active site. The substrate specificity of TNMT enzymes appears to arise from the arrangement of subgroup-specific stereospecific recognition elements relative to catalytic elements that are more widely-conserved among all BIA NMTs. The binding mode of protoberberine compounds to GfTNMT appears to be similar to coclaurine NMT, with the isoquinoline rings buried deepest in the binding pocket. This binding mode differs from that of pavine NMT, in which the benzyl ring is bound more deeply than the isoquinoline rings. The insights into substrate recognition and catalysis provided here form a sound basis for the rational engineering of NMT enzymes for chemoenzymatic synthesis and metabolic engineering.

Motor control dysfunction persisting during symptom remission in persons with recurrent low back pain (LBP) may contribute to the recurrence of pain.To investigate trunk control in persons in remission from recurrent LBP and in back-healthy controls using a dynamic, internally driven balance task. No differences in task performance were expected between groups, but it was hypothesized that persons with recurrent LBP would exhibit greater trunk coupling, consistent with a trunk-stiffening strategy.In this cross-sectional controlled laboratory study, persons with and without recurrent LBP (n = 19 per group) completed the balance-dexterity task, which involved balancing on one limb in standing while compressing an unstable spring with the other. Task performance measures included center-of-pressure velocity under the stance limb and vertical force variability under the spring. Trunk coupling was quantified with the coefficient of determination (There were no differences between groups for any task performance measure. The group in remission from recurrent LBP exhibited reduced trunk coupling, or more dissociated thorax and pelvis motion, compared to the healthy control group (The balance-dexterity task is a submaximal, internally driven unstable balance task during which more dissociated trunk motion was observed in persons in remission from recurrent LBP. Findings underscore the task-dependent nature of trunk control research and assessment in persons with recurrent LBP.

Background Tendinopathy of the flexor hallucis longus, commonly called “dancer's tendinitis”, is a prevalent injury among female ballet dancers. Limited success of non-surgical interventions leads to many dancers undergoing tenolysis surgeries with risks and recovery times that can be detrimental to a dance career. The purpose of this study was to evaluate lower limb kinematics and muscle coordination during a modified traditional heel raise exercise where the toes hang off the edge of the support surface. We hypothesized this would decrease activation of the flexor hallucis longus and increase activation of larger plantarflexors. Methods Healthy non-dancers (n = 11), healthy dancers (n = 10), and dancers with flexor hallucis longus tendinopathy (n = 9) performed traditional (“toes-on”) and modified (“toes-off”) heel raises with kinematic and electromyographic instrumentation of the lower leg. Findings Participants maintained ankle excursion with the toes-off modification, while metatarsophalangeal joints had reduced excursion and greater excursion variability. Most healthy dancers (9/10) decreased flexor hallucis longus activation as predicted, but dancers with flexor hallucis longus tendinopathy showed a variable response with some decreasing activation (3/9) but others increasing activation up to 4-times. There were no changes in activation of other plantarflexors. Across groups, intrinsic foot muscle activation decreased with the toes-off modification. Interpretation The toes-off modification decreased flexor hallucis longus activation in most of the healthy dancers but was insufficient to shift muscle coordination from the flexor hallucis longus to larger plantarflexors in dancers with flexor hallucis longus tendinopathy. Future work should investigate clinical cues or modifications to this “toes-off” heel raise intervention.

Background Brachial plexus birth palsy occurs in 0.4 to 4.6 of every 1000 live births, with residual shoulder dysfunction in approximately one third of cases. Clinical measures, such as the Mallet classification, provide no insight into the scapulothoracic and glenohumeral contributions to tested global shoulder movements. This study describes the scapulothoracic and glenohumeral components of shoulder motion during the modified Mallet test. Methods Twelve children with Erb’s palsy (C5-6) and 8 children with extended Erb’s palsy (C5-7) were recruited. The unaffected limbs of 6 subjects were also tested. Locations of markers placed on the thorax, humerus, and scapula were recorded in a neutral position and each of the modified Mallet positions. Scapulothoracic, glenohumeral, and humerothoracic helical displacements and acromion process linear displacements were compared between groups. Results The brachial plexus birth palsy groups exhibited significantly smaller glenohumeral displacements in all modified Mallet positions and significantly larger scapulothoracic displacements in the global external rotation and hand to mouth positions. Discriminant function analysis using only humerothoracic variables correctly classified 76.9% of subjects. Discriminant function analysis incorporating scapulothoracic, glenohumeral, and acromion process displacement variables produced accuracy of 92.6%. Conclusions Children with brachial plexus birth palsy demonstrated decreased glenohumeral contributions to achieve every modified Mallet position and increased scapulothoracic contribution in two positions compared with the unaffected group. Different scapulothoracic and glenohumeral strategies were identified between groups. Finally, scapulothoracic and glenohumeral components of shoulder motion are more specific than humerothoracic measures to diagnostic classification.

Several strands of empirical work have begun to focus on how institutional change and the actions of institutional actors occurring in one locale can impact organizational behaviour in another. In this paper, we explore how the strategic decision making on the part of supplier firms is shaped by extra-jurisdictional regulative and normative pressures affecting their potential clients. The study’s context is the adoption of practices by suppliers to an industry supported by regulation endorsing private voluntary standards. A novel dataset is utilised which tracks the adoption of green building practices by a panel of 226 architecture studios in Australia from 2008-2015. In line with our theoretical predictions, firms are found to respond to regulation and social norms in jurisdictions where they do not operate but where their prospective clients do. In their responses to extra-jurisdictional regulation, firms are found to vary to an extent that depends on a firm’s relative power, as defined by its embeddness vis-a- vis its potential clients in an emerging market. These findings suggest that the behavioural effects of institutions can diffuse across contexts through the exchange relations between firms. As institutional effects diffuse through firms’ relations, they are conditioned by structural properties of those exchange relations.

A series of macrocyclic compounds containing a cyclic constraint in the P2–P4 linker region have been discovered and shown to exhibit excellent HCV NS3/4a genotype 3a and genotype 1b R155 K, A156T, A156 V, and D168 V mutant activity while maintaining high rat liver exposure. The effect of the constraint is most dramatic against gt 1b A156 mutants where ∼20-fold improvements in potency are achieved by introduction of a variety of ring systems into the P2–P4 linker.

The Hedgehog (Hh-) signalling pathway is a key developmental pathway and there is a growing body of evidence showing that this pathway is aberrantly reactivated in a number of human tumors. Novel agents capable of inhibiting this pathway are sought, and an entirely novel series of smoothened (Smo) antagonists capable of inhibiting the pathway have been identified through uHTS screening. Extensive exploration of the scaffold identified the key functionalities necessary for potency, enabling potent nanomolar Smo antagonists like 91 and 94 to be developed. Optimization resulted in the most advanced compounds displaying low serum shift, clean off-targets profile, and moderate clearance in both rats and dogs. These compounds are valuable tools with which to probe the biology of the Hh-pathway.

Smoothened (Smo) antagonists are emerging as new therapies for the treatment of neoplasias with aberrantly reactivated hedgehog (Hh) signaling pathway. A novel series of 4-[3-(quinolin-2-yl)-1,2,4-oxadiazol-5-yl]piperazinyl ureas as smoothened antagonists was recently described, herein the series has been further optimized through the incorporation of a basic amine into the urea. This development resulted in identification of some exceptionally potent smoothened antagonists with low serum shifts, however, reductive ring opening on the 1,2,4-oxadiazole in rats limits the applicability of these compounds in in vivo studies.

Raltegravir is the first integrase strand transfer inhibitor approved for the treatment of HIV-1 infection. As the first agent in this new class of antiretroviral therapies, raltegravir has demonstrated safety and efficacy in treatment-naive as well as heavily pretreated HIV-infected patients failing therapy with multidrug-resistant virus. Raltegravir has a favorable drug interaction profile that permits both administration to a wide, demographically diverse patient population and coadministration with many other therapeutic agents, including antiretroviral agents and supportive medications, without restrictions or dose adjustment. Data through 96 weeks of follow-up in three phase III studies, protocol 021 (STARTMRK) in treatment-naive patients, and protocols 018 (BENCHMRK-1) and 019 (BENCHMRK-2) in treatment-experienced patients, demonstrated the potent and durable antiretroviral and immunologic effects and the favorable long-term safety profile of raltegravir in both treatment-naive and treatment-experienced patients. Raltegravir represents an important addition to the current armamentarium for the treatment of HIV infection.

A primary objective of pharmacokinetic-pharmacodynamic (PKPD) reasoning is to identify key in vivo drug and system proper¬ties, enabling prediction of the magnitude and time course of drug responses under physiological and pathological conditions in animals and man. Since the pharmacological response generated by a drug is highly dependent on the actual system used to study its action, knowledge about its potency and efficacy at a given concentration or dose is insufficient to obtain a proper understanding of its pharmacodynamic profile. Hence, the output of PKPD activities extends beyond the provision of quantitative measures (models) of results, to the design of future protocols. Furthermore, because PKPD integrates DMPK (e.g. clearance) and pharmacology (e.g. potency),it provides an anchor point for compound selection, and, as such, should be viewed as an important weapon in medicinal chemistry. Here we outline key PK concepts relevant to PD, and then consider real-life experiments to illustrate the importance to the medicinal chemist of data obtained by PKPD. Useful assumptions and potential pitfalls are described, providing a holistic view of the plethora of determinants behind in vitro-in vivo correlations. By condensing complexity to simplicity, there are not only consequences for experimental design, and for the ranking and design of compounds, but it is also possible to make important predictions such as the impact of changes in drug potency and kinetics. In short, by using quantitative methods to tease apart pharmacodynamic complexities such as temporal differences and changes in plasma protein binding, it is possible to target the changes necessary for improving a compound's profile.

Infections caused by hepatitis C virus (HCV) are a significant world health problem for which novel therapies are in urgent demand. The polymerase of HCV is responsible for the replication of viral genome and has been a prime target for drug discovery efforts. Here, we report on the further development of tetracyclic indole inhibitors, binding to an allosteric site on the thumb domain. Structure-activity relationship (SAR) studies around an indolo-benzoxazocine scaffold led to the identification of compound 33 (MK-3281), an inhibitor with good potency in the HCV subgenomic replication assay and attractive molecular properties suitable for a clinical candidate. The compound caused a consistent decrease in viremia in vivo using the chimeric mouse model of HCV infection.

It is well-documented that regulative institutions, comprised of administrative guidelines, rules and laws (Scott, 1995), have the power to shape firm behaviour. In recent years, researchers have b...

The purpose of this study was to investigate the in vivo absorption enhancement of a nucleoside (phosphoramidate prodrug of 2'-methyl-cytidine) anti-viral agent of proven efficacy by means of intestinal permeation enhancers. Natural nucleosides are hydrophilic molecules that do not rapidly penetrate cell membranes by diffusion and their absorption relies on specialized transporters. Therefore, the oral absorption of nucleoside prodrugs and the target organ concentration of the biologically active nucleotide can be limited due to poor permeation across the intestinal epithelium. In the present study, the specificity, concentration dependence, and effect of four classes of absorption promoters, i.e. fatty acids, steroidal detergents, mucoadhesive polymers, and secretory transport inhibitors, were evaluated in a rat in vivo model. Sodium caprate and alpha-tocopheryl-polyethyleneglycol-1000-succinate (TPGS) showed a significant effect in increasing liver concentration of nucleotide (5-fold). These results suggested that both excipients might be suited in a controlled release matrix for the synchronous release of the drug and absorption promoter directly to the site of absorption and highlights that the effect is strictly dependent on the absorption promoter dose. The feasibility of such a formulation approach in humans was evaluated with the aim of developing a solid dosage form for the peroral delivery of nucleosides and showed that these excipients do provide a potential valuable tool in pre-clinical efficacy studies to drive discovery programs forward.

An efficient and general synthesis of homochiral alpha-amino substituted bromo-heteroaromatics B using a diastereoselective 1,2-addition has been developed. The obtained heteroaromatic intermediates allow for a rapid two-dimensional exploration of a new series of histone deacetylase inhibitors, through Suzuki-Miyaura cross-coupling reactions for the introduction of a second aromatic element, followed by global deprotection and derivatization of the amino group. (C) 2009 Published by Elsevier Ltd.

Infections caused by the hepatitis C virus (HCV) are a significant world health problem for which novel therapies are in urgent demand. The NS5B polymerase of HCV is responsible for the replication of viral RNA and has been a prime target in the search for novel treatment options. We had discovered allosteric finger-loop inhibitors based on a thieno[3,2-b]pyrrole scaffold as an alternative to the related indole inhibitors. Optimization of the thienopyrrole series led to several N-acetamides with submicromolar potency in the cell-based replicon assay, but they lacked oral bioavailability in rats. By linking the N4-position to the ortho-position of the C5-aryl group, we were able to identify the tetracyclic thienopyrrole 40, which displayed a favorable pharmacokinetic profile in rats and dogs and is equipotent with recently disclosed finger-loop inhibitors based on an indole scaffold.

The currently approved treatment for hepatitis C virus infections is a combination of Ribavirin and pegylated Interferon. It leads to a sustained virologic response in approximately only half of the patients treated. For this reason there is an urgent need of new therapeutic agents. 2'-C-Methylcytidine is the first nucleoside inhibitor of the HCV NS5B polymerase that was efficacious in reducing the viral load in patients infected with HCV. The application of a monophosphate prodrug approach based on unprecedented cyclic phosphoramidates is reported. Our SAR studies led to compounds that are efficiently converted to the active triphosphate in human hepatocytes.

PARP inhibitors have been demonstrated to retard intracellular DNA repair and therefore sensitize tumor cells to cytotoxic agents or ionizing radiation. We report the identification of a novel class of PARP1 inhibitors, containing a pyrrolo moiety fused to a dihydroisoquinolinone, derived from virtual screening of the proprietary collection. SAR exploration around the nitrogen of the aminoethyl appendage chain of 1 led to compounds that displayed low nanomolar activity in a PARP1 enzymatic assay.

The optimization of a potent, class I selective ketone HDAC inhibitor is shown. It possesses optimized pharmacokinetic properties in preclinical species, has a clean off-target profile, and is negative in a microbial mutagenicity (Ames) test. In a mouse xenograft model it shows efficacy comparable to that of vorinostat at a 10-fold reduced dose.

A variety of new prodrugs of 2'-methyl cytidine based on acyloxy ethylamino phosphoramidates have been synthesized and tested in vitro and in vivo for their biological activity. Compared with the parent drug a 10- to 20-fold increase in formation of nucleotide triphosphate in rat and human hepatocytes could be achieved.

The RNA replication machinery of HCV is a multi-subunit membrane–associated complex. NS5A has emerged as an active component of HCV replicase, possibly involved in regulation of viral replication and resistance to the antiviral effect of interferon. We report here substituted piperazinyl- N -(aryl)benzamides as potent inhibitors of HCV replication exerted via modulation of the dimerization of NS5A.

5-(Trifluoroacetyl)thiophene-2-carboxamides were found to be potent and selective class II HDAC inhibitors. This paper describes their further development and the investigation on the cause for the lack of cell-based activity. A rapid screening assay was set up which enabled the identification of more metabolic stable compounds as potent and selective class II HDAC inhibitors.

Histone deacetylase (HDAC) inhibitors offer a promising strategy for cancer therapy and the first generation HDAC inhibitors are currently in the clinic. Herein we describe the optimization of a series of ketone small molecule HDAC inhibitors leading to potent and selective class I HDAC inhibitors with good dog PK.

The identification of class II HDAC inhibitors has been hampered by lack of efficient enzyme assays, in the preceding paper two assays have been developed to improve the efficiency of these enzymes: mutating an active site histidine to tyrosine, or by the use of a trifluoroacetamide lysine substrate, allowing screening to identify class II HDAC inhibitors. Herein, 2-trifluoroacetylthiophenes have been demonstrated to inhibit class II HDACs, resulting in the development of a series of 5-(trifluoroacetyl)thiophene-2-carboxamides as novel, potent and selective class II HDAC inhibitors. X-ray crystal structures of the HDAC 4 catalytic domain with a bound inhibitor demonstrate these compounds are active site inhibitors and bind in their hydrated form.

Poly(ADP-ribose) polymerases (PARPs) are involved in DNA repair following damage by endogenous or exogenous processes. It has become clear over the past decade that inhibition of PARP in the context of defects in other DNA repair mechanisms provide a tumor specific way to kill cancer cells. We describe the rationale for this approach and the design and discovery of niraparib, a potent PARP-1/2 inhibitor with good cell based activity, selectivity for cancer over normal cells, and oral bioavailability. Niraparib was characterized in a number of preclinical models before moving to phase I clinical trials, where it showed excellent human pharmacokinetics suitable for once a day oral dosing, achieved its pharmacodynamic target for PARP inhibition, and had promising activity in cancer patients. It is currently being tested in phase 3 clinical trials as maintenance therapy in ovarian cancer and as a treatment for breast cancer.

The hepatitis C virus (HCV) polymerase is required for replication of the viral genome and is a key target for therapeutic intervention against HCV. We have determined the crystal structures of the HCV polymerase complexed with two indole-based allosteric inhibitors at 2.3- and 2.4-Angstroms resolution. The structures show that these inhibitors bind to a site on the surface of the thumb domain. A cyclohexyl and phenyl ring substituents, bridged by an indole moiety, fill two closely spaced pockets, whereas a carboxylate substituent forms a salt bridge with an exposed arginine side chain. Interestingly, in the apoenzyme, the inhibitor binding site is occupied by a small alpha-helix at the tip of the N-terminal loop that connects the fingers and thumb domains. Thus, these molecules inhibit the enzyme by preventing formation of intramolecular contacts between these two domains and consequently precluding their coordinated movements during RNA synthesis. Our structures identify a novel mechanism by which a new class of allosteric inhibitors inhibits the HCV polymerase and open the way to the development of novel antiviral agents against this clinically relevant human pathogen.

Infections caused by hepatitis C virus (HCV) are a significant world health problem for which novel therapies are in urgent demand. Compounds that block replication of subgenomic HCV RNA in liver cells are of interest because of their demonstrated antiviral effect in the clinic. In followup to our recent report that indole-N-acetamides (e.g., 1) are potent allosteric inhibitors of the HCV NS5B polymerase enzyme, we describe here their optimization as cell-based inhibitors. The crystal structure of 1 bound to NS5B was a guide in the design of a two-dimensional compound array that highlighted that formally zwitterionic inhibitors have strong intracellular potency and that pregnane X receptor (PXR) activation (an undesired off-target activity) is linked to a structural feature of the inhibitor. Optimized analogues devoid of PXR activation (e.g., 55, EC(50) = 127 nM) retain strong cell-based efficacy under high serum conditions and show acceptable pharmacokinetics parameters in rat and dog.

Allosteric inhibition of the hepatitis C virus (HCV) NS5B RNA-dependent RNA polymerase enzyme has recently emerged as a viable strategy toward blocking replication of viral RNA in cell-based systems. We report here a novel class of allosteric inhibitor of NS5B that shows potent affinity for the NS5B enzyme and effective inhibition of subgenomic HCV RNA replication in HUH-7 cells. Inhibitors from this class have promising characteristics for further development as anti-HCV agents.

HCV NS3/4a protease inhibitors are proven therapeutic agents against chronic hepatitis C virus infection, with boceprevir and telaprevir having recently received regulatory approval as add-on therapy to pegylated interferon/ribavirin for patients harboring genotype 1 infections. Overcoming antiviral resistance, broad genotype coverage, and a convenient dosing regimen are important attributes for future agents to be used in combinations without interferon. In this communication, we report the preclinical profile of MK-5172, a novel P2-P4 quinoxaline macrocyclic NS3/4a protease inhibitor currently in clinical development. The compound demonstrates subnanomolar activity against a broad enzyme panel encompassing major hepatitis C virus (HCV) genotypes as well as variants resistant to earlier protease inhibitors. In replicon selections, MK-5172 exerted high selective pressure, which yielded few resistant colonies. In both rat and dog, MK-5172 demonstrates good plasma and liver exposures, with 24-h liver levels suggestive of once-daily dosing. When administered to HCV-infected chimpanzees harboring chronic gt1a or gt1b infections, MK-5172 suppressed viral load between 4 to 5 logs at a dose of 1 mg/kg of body weight twice daily (b.i.d.) for 7 days. Based on its preclinical profile, MK-5172 is anticipated to be broadly active against multiple HCV genotypes and clinically important resistance variants and highly suited for incorporation into newer all-oral regimens.

A series of tricyclic pyridones has been evaluated as benzodiazepine site ligands with functional selectivity for the alpha(3) over the alpha(1) containing subtype of the human GABA(A) receptor ion channel. This investigation led to the identification of a high affinity, functionally selective, orally bioavailable benzodiazepine site ligand that demonstrated activity in rodent anxiolysis models and reduced sedation relative to diazepam.

The RNA-dependent RNA polymerase of hepatitis C virus (HCV) is the catalytic subunit of the viral RNA amplification machinery and is an appealing target for the development of new therapeutic agents against HCV infection. Nonnucleoside inhibitors based on a benzimidazole scaffold have been recently reported. Compounds of this class are efficient inhibitors of HCV RNA replication in cell culture, thus providing attractive candidates for further development. Here we report the detailed analysis of the mechanism of action of selected benzimidazole inhibitors. Kinetic data and binding experiments indicated that these compounds act as allosteric inhibitors that block the activity of the polymerase prior to the elongation step. Escape mutations that confer resistance to these compounds map to proline 495, a residue located on the surface of the polymerase thumb domain and away from the active site. Substitution of this residue is sufficient to make the HCV enzyme and replicons resistant to the inhibitors. Interestingly, proline 495 lies in a recently identified noncatalytic GTPbinding site, thus validating it as a potential allosteric site that can be targeted by small-molecule inhibitors of HCV polymerase. Hepatitis C virus (HCV) is the causative agent of the majority of chronic liver disease throughout the world. More than 170 million individuals are estimated to be infected with this virus (27). The size of the HCV epidemic and the limited efficacy of current therapy (based on the use of alpha interferon) have stimulated intense research efforts towards the development of antiviral drugs that are both better tolerated and more effective. The most widely established strategy for developing novel anti-HCV therapeutics aims at the identification of low-molecular-weight inhibitors of essential HCV enzymes.