Ferrocene-based inhibitors of hepatitis C virus replication that target NS5A with low picomolar in vitro antiviral activity

An unprecedented series of organometallic HCV (hepatitis C virus) NS5A (nonstructural 5A protein) replication complex inhibitors that incorporates a 1,1′-ferrocenediyl scaffold was explored. This scaffold introduces the elements of linear flexibility and non-planar topology that are unconventional for this class of inhibitors. Data from 2-D NMR spectroscopic analyses of these complexes in solution support an anti (unstacked) arrangement of the pharmacophoric groups. Several complexes demonstrate single-digit picomolar in vitro activity in an HCV genotype-1b replicon system. One complex to arise from this investigation (10a) exhibits exceptional picomolar activity against HCV genotype 1a and 1b replicons, low hepatocellular cytotoxicity, and good pharmacokinetic properties in rat.     

Estimating the effective clearance rate and refiltration by zebra mussels, Dreissena polymorpha, in a stratified reservoir

1. The grazing impact of zebra mussels, Dreissena polymorpha Pallas, is often evaluated by applying the individual filtration rate measured in the laboratory to the field abundance and then by comparing the total volume of water filtered with the whole lake volume. Since this approach overlooks refiltration, it overestimates the grazing impact of zebra mussels. To deal with this problem, the present authors developed an in situ method for collecting faeces and pseudofaeces to measure the actual volume of water that is cleared of suspended particles by Dreissena in a unit time under a given set of environmental conditions. This is termed the effective clearance rate (ECR). 2. The experiment was conducted in Hargus Lake, OH, U.S.A., a small thermally stratified reservoir, to test the effects of spatial aggregation, mussel density and the concentration of particulate inorganic matter (PIM) on the effective clearance rate of Dreissena. 3. Over 40 measurements, the ECR values ranged from 15.3 to 68.6 mL ind^??1 h^??1. Much of the variation can be explained by colony form, mussel density and seston concentration. The effects of these variables were all statistically significant. The average ECR for isolated individuals was higher than that for those in clumps (40.4 versus 32.8 mL ind^??1 h^??1), which is attributed to increased refiltration in the cores of the clumps. The ECR decreased with increased zebra mussel density because of intensified competition for food particles within the group. The ECR increased with increased PIM concentration in the lake water, which may be interpreted as a result of enhanced water mixing which ultimately caused increases in both sediment resuspension and particle delivery to the mussels. 4. Taking the filtration rates for a 20-mm mussel to be between 116 and 234 mL ind^??1 h^??1, based on data from the literature, the clumped mussels under the present experimental conditions would have a refiltration ratio between 3.4 and 6.9. 5. The present authors developed an areal clearance model which predicts that seston removal by the Dreissena population is limited by the particle delivery from the ambient water to the mussel bed and will reach a maximum value beyond which no further increase will occur with increased population density. 6. It is concluded that the direct grazing impact of zebra mussel on phytoplankton in thermally stratified lakes is much less effective than predicted from simple filtration rate estimation.     

VDAC1 serves as a mitochondrial binding site for hexokinase in oxidative muscles

Voltage-dependent anion channels (VDACs), also known as mitochondrial porins, are the main pathway for metabolites across the mitochondrial outer membrane and may serve as binding sites for kinases, including hexokinase. We determined that mitochondria-bound hexokinase activity is significantly reduced in oxidative muscles (heart and soleus) in vdac1^−/− mice. The activity data were supported by western blot analysis using HK2 specific antibody. To gain more insight into the physiologic mean of the results with the activity data, VDAC deficient mice were subjected to glucose tolerance testing and exercise-induced stress, each of which involves tissue glucose uptake via different mechanisms. vdac1^−/− mice exhibit impaired glucose tolerance whereas vdac3^−/− mice have normal glucose tolerance and exercise capacity. Mice lacking both VDAC1 and VDAC3 (vdac1^−/−/vdac3^−/−) have reduced exercise capacity together with impaired glucose tolerance. Therefore, we demonstrated a link between VDAC1 mediated mitochondria-bound hexokinase activity and the capacity for glucose clearance.     

内蒙古地产紫沙参多糖免疫功能

给小鼠灌胃口服紫沙参多糖 ４００ ｍｇ· ｋｇ－１、８００ ｍｇ· ｋｇ－１,观察其连续给药对免疫功能的影响。结果显示：在５ｄ后能显著地增加小鼠耳肿胀度,提示紫沙参多糖能够增强二硝基氯苯诱导的小鼠迟发性变态反应（ＤＴＨ）;在７ ｄ后能显著地增加碳粒廓清指数Ｋ和吞噬指数α,增强单核巨噬细胞的吞噬功能,能显著地增加免疫器官胸腺、脾脏重量,增强机体免疫作用。试验采用国家中药二类新药云芝多糖 １０００ ｍｇ· ｋｇ－１作为阳性对照组。     

The discovery of benzoxazine sulfonamide inhibitors of NaV1.7: Tools that bridge efficacy and target engagement

The voltage-gated sodium channel Na_V1.7 has received much attention from the scientific community due to compelling human genetic data linking gain- and loss-of-function mutations to pain phenotypes. Despite this genetic validation of Na_V1.7 as a target for pain, high quality pharmacological tools facilitate further understanding of target biology, establishment of target coverage requirements and subsequent progression into the clinic. Within the sulfonamide class of inhibitors, reduced potency on rat Na_V1.7 versus human Na_V1.7 was observed, rendering in vivo rat pharmacology studies challenging. Herein, we report the discovery and optimization of novel benzoxazine sulfonamide inhibitors of human, rat and mouse Na_V1.7 which enabled pharmacological assessment in traditional behavioral rodent models of pain and in turn, established a connection between formalin-induced pain and histamine-induced pruritus in mice. The latter represents a simple and efficient means of measuring target engagement.     

Pharmacokinetic de-risking tools for selection of monoclonal antibody lead candidates

Pharmacokinetic studies play an important role in all stages of drug discovery and development. Recent advancements in the tools for discovery and optimization of therapeutic proteins have created an abundance of candidates that may fulfill target product profile criteria. Implementing a set of in silico, small scale in vitro and in vivo tools can help to identify a clinical lead molecule with promising properties at the early stages of drug discovery, thus reducing the labor and cost in advancing multiple candidates toward clinical development. In this review, we describe tools that should be considered during drug discovery, and discuss approaches that could be included in the pharmacokinetic screening part of the lead candidate generation process to de-risk unexpected pharmacokinetic behaviors of Fc-based therapeutic proteins, with an emphasis on monoclonal antibodies.     

Effects of glyphosate herbicide on soil and litter macro-arthropods in rainforest: Implications for forest restoration

Summary    Ecological restoration activities, including reforestation, often involve the use of herbicides for the removal of weedy plant cover. Little is known, however, about the effects of herbicides on assemblages of non-target organisms that colonize restored patches. We describe a field experiment to investigate effects of glyphosate herbicide (Roundup® Biactive™) on rainforest-associated soil- and litter-dwelling macro-arthropods. Our experimental protocol differed in two ways from other ecotoxicological studies of herbicides. First, we applied herbicide at a rate considerably greater than the manufacturer's recommended maximum in order to simulate worst-case scenarios that may occur in the practice of forest restoration. Second, our field experiment was carried out under dense canopy cover with sparse understorey vegetation, so that indirect impacts caused by the loss of existing vegetation were eliminated. Paired herbicide-treated and control plots were created within five rainforest remnants on the Maleny plateau of subtropical eastern Australia. Macro-arthropods were collected using litter extraction before, approximately 3 days after, and 3 months after herbicide application. Responses of arthropods were analysed at two levels of taxonomic resolution: 'coarse' arthropods (arthropods sorted to Order/Class), and ant species. Our results suggest that the use of glyphosate herbicide formulated as Roundup® Biactive™ is suitable for the control of unwanted plants in rainforest restoration sites as it appears to have minimal impact on assemblages of soil and litter macro-arthropods or at least those typical of intact rainforest.     

Functional analysis of the Drosophila immune response during aging

One of the most dramatic changes associated with aging involves immunity. In aging mammals, immune function declines and chronic inflammation develops. The biological significance of this phenomenon and its relationship with aging is a priority for aging research. Drosophila is an invaluable tool in understanding the effects of aging on the immune response. Similar to the state of chronic inflammation in mammals, Drosophila exhibits a drastic up-regulation of immunity-related genes with age. However, it remains unclear whether immune function declines with age as seen in mammals. We evaluated the impact of aging on Drosophila immune function by examining across age the ability to eliminate and survive different doses of bacterial invaders. Our findings show that aging reduces the capacity to survive a bacterial infection. In contrast, we found no evidence that aging affects the ability to eliminate bacteria indicating that the mechanisms underlying immune senescence are not involved in eliminating bacteria or preventing their proliferation.     

Persistent migration of neuroblasts from the subventricular zone to the injured striatum mediated by osteopontin following intracerebral hemorrhage

We investigated intracerebral hemorrhage (ICH)-induced lateral migration of neuroblasts and the mechanism underlying this migration. ICH model was induced by collagenase injection into the striatum of adult wild-type and osteopontin (OPN) knockout mice. In the wild-type mice, the lateral migration of neuroblasts from the ipsislateral subventricular zone (SVZ) towards the hematoma started at day 3 and continued up to day 28 after ICH. In addition to migrating towards the hematoma, neuroblasts also migrated to the area of ipsilateral striatum remote to the hematoma. The migrating neuroblasts were closely associated with activated astrocytes and blood vessels in the injured striatum. Following ICH, the expression of OPN was up-regulated in the ipsilateral striatum from day 1 to day 28. In vitro , OPN treatment did not affect the proliferation of neural progenitors, but enhanced the trans-well and radial migration of neural progenitors. In vivo , OPN deficiency did not affect the proliferation of neural progenitors in the SVZ. However, following ICH a significant decrease in lateral neuroblast migration was observed in the OPN knockout mice compared with the wild-type mice. These results suggest that increased OPN expression in the injured striatum plays a significant role in the lateral migration of neuroblasts following ICH.