Successful drug development despite adverse preclinical findings part 1: processes to address issues and most important findings

Unexpected adverse preclinical findings (APFs) are not infrequently encountered during drug development. Such APFs can be functional disturbances such as QT prolongation, morphological toxicity or carcinogenicity. The latter is of particular concern in conjunction with equivocal genotoxicity results. The toxicologic pathologist plays an important role in recognizing these effects, in helping to characterize them, to evaluate their risk for man, and in proposing measures to mitigate the risk particularly in early clinical trials. A careful scientific evaluation is crucial while termination of the development of a potentially useful drug must be avoided. This first part of the review discusses processes to address unexpected APFs and provides an overview over typical APFs in particular classes of drugs. If the mode of action (MoA) by which a drug candidate produces an APF is known, this supports evaluation of its relevance for humans. Tailor-made mechanistic studies, when needed, must be planned carefully to test one or several hypotheses regarding the potential MoA and to provide further data for risk evaluation. Safety considerations are based on exposure at no-observed-adverse-effect levels (NOAEL) of the most sensitive and relevant animal species and guide dose escalation in clinical trials. The availability of early markers of toxicity for monitoring of humans adds further safety to clinical studies. Risk evaluation is concluded by a weight of evidence analysis (WoE) with an array of parameters including drug use, medical need and alternatives on the market. In the second part of this review relevant examples of APFs will be discussed in more detail.     

Comparison of chemistry analytes between 2 portable, commercially available analyzers and a conventional laboratory analyzer in reptiles

Background: Advantages of handheld and small bench‐top biochemical analyzers include requirements for smaller sample volume and practicality for use in the field or in practices, but little has been published on the performance of these instruments compared with standard reference methods in analysis of reptilian blood. Objective: The aim of this study was to compare reptilian blood biochemical values obtained using the Abaxis VetScan Classic bench‐top analyzer and a Heska i‐STAT handheld analyzer with values obtained using a Roche Hitachi 911 chemical analyzer. Methods: Reptiles, including 14 bearded dragons (Pogona vitticeps), 4 blue‐tongued skinks (Tiliqua gigas), 8 Burmese star tortoises (Geochelone platynota), 10 Indian star tortoises (Geochelone elegans), 5 red‐tailed boas (Boa constrictor), and 5 Northern pine snakes (Pituophis melanoleucus melanoleucus), were manually restrained, and a single blood sample was obtained and divided for analysis. Results for concentrations of albumin, bile acids, calcium, glucose, phosphates, potassium, sodium, total protein, and uric acid and activities of aspartate aminotransferase and creatine kinase obtained from the VetScan Classic and Hitachi 911 were compared. Results for concentrations of chloride, glucose, potassium, and sodium obtained from the i‐STAT and Hitachi 911 were compared. Results: Compared with results from the Hitachi 911, those from the VetScan Classic and i‐STAT had variable correlations, and constant or proportional bias was found for many analytes. Bile acid data could not be evaluated because results for 44 of 45 samples fell below the lower linearity limit of the VetScan Classic. Conclusions: Although the 2 portable instruments might provide measurements with clinical utility, there were significant differences compared with the reference analyzer, and development of analyzer‐specific reference intervals is recommended.     

Reprogramming cellular behavior with RNA controllers responsive to endogenous proteins

Synthetic genetic devices that interface with native cellular pathways can be used to change natural networks to implement new forms of control and behavior. The engineering of gene networks has been limited by an inability to interface with native components. We describe a class of RNA control devices that overcome these limitations by coupling increased abundance of particular proteins to targeted gene expression events through the regulation of alternative RNA splicing. We engineered RNA devices that detect signaling through the nuclear factor κB and Wnt signaling pathways in human cells and rewire these pathways to produce new behaviors, thereby linking disease markers to noninvasive sensing and reprogrammed cellular fates. Our work provides a genetic platform that can build programmable sensing-actuation devices enabling autonomous control over cellular behavior.     

ANXITANE® tablets reduce fear of human beings in a laboratory model of anxiety-related behavior

Anxiety and fear are common underlying factors in many canine behavior problems that impair the human–pet bond and often result in abandonment, relinquishment, or euthanasia. A combination of behavioral and pharmacological interventions is used to ameliorate the behavioral signs associated with anxiety-related behaviors in dogs, but there continues to be need for effective interventions. The current study examined the effects of the nutraceutical ANXITANE^® (l-Theanine) chewable tablets on fear of unfamiliar human beings. We first characterized dogs as anxious on the basis of the existence of a fear response to human beings in their home-pen. We then demonstrated that dogs characterized as anxious (N = 10) showed reduced interaction with an unknown human being as compared with normal controls (N = 7). The effect of an administration of ANXITANE^® tablets (N = 5) on these anxious Beagle dogs was compared with placebo (N = 5). Objective behavioral measures of anxiety were obtained using an open-field test, a human interaction test, and an actiwatch protocol that allowed monitoring of activity over 24-hours. The ANXITANE^® tablets-treated dogs showed greater human interaction and approach than the placebo control group, and no side effects related to treatment, including motor stimulant or sedative effects, were seen. The current study suggests that ANXITANE^® tablets are effective for reducing fearful behavior toward unfamiliar human beings in dogs and supports their use for treating anxiety-related behaviors.     

Comparison of Computed Tomographic and Radiographic Popliteal Lymphangiography in Normal Dogs

Objective: To (1) describe computed tomographic (CT) popliteal lymphangiography; (2) compare the number of thoracic duct (TD) branches detected by CT and by radiography after popliteal lymphangiography; and (3) to compare the number of branches detected after left and right popliteal lymphangiography. Study Design: Experimental study. Animals: Adult dogs (n=6). Methods: A randomly selected popliteal lymph node was percutaneously injected with 12 mL iodinated contrast medium through a 25‐g butterfly catheter over 4–5 minutes. Lateral and ventrodorsal (VD) thoracic radiograph projections and thoracic CT were performed. The procedure was repeated using the contralateral lymph node after a 48–72 hours washout period. Results: One dog had TD branches visible on CT but not on radiographs. A significantly greater number of TD branches were observed with CT popliteal lymphangiography compared with lateral and VD radiographic popliteal lymphangiography (P=.003 and P<.001, respectively). The number of visible TD branches observed between the 6th thoracic and 1st lumbar vertebrae were not significantly different in these dogs (P=.146). A significant difference in number of TD branches observed was not found after left or right popliteal lymph node injection (P=.097). Conclusions: CT popliteal lymphangiography consistently identified a greater number of TD branches when compared with radiographic popliteal lymphangiography. Injection of either popliteal lymph node resulted in the same number of TD branches being observed.     

Heavy Metal Concentration Survey in Soils and Plants of the Les Malines Mining District (Southern France): Implications for Soil Restoration

Mining activities generate spoils and effluents with extremely high metal concentrations of heavy metals that might have adverse effects on ecosystems and human health. Therefore, information on soil and plant metal concentrations is needed to assess the severity of the pollution and develop a strategy for soil reclamation such as phytoremediation. Here, we studied soils and vegetation in three heavily contaminated sites with potential toxic metals and metalloids (Zn, Pb, Cd, As, TI) in the mining district of Les Malines in the Languedoc region (southern France). Extremely high concentrations were found at different places such as the Les Aviniéres tailing basins (up to 160,000 mg kg^?C1 Zn, 90,000 mg kg^?C1 Pb, 9,700 mg kg^?C1 of As and 245 mg kg^?C1 of Tl) near a former furnace. Metal contamination extended several kilometres away from the mine sites probably because of the transport of toxic mining residues by wind and water. Spontaneous vegetation growing on the three mine sites was highly diversified and included 116 plant species. The vegetation cover consisted of species also found in non-contaminated soils, some of which have been shown to be metal-tolerant ecotypes (Festuca arvernensis, Koeleria vallesiana and Armeria arenaria) and several Zn, Cd and Tl hyperaccumulators such as Anthyllis vulneraria, Thlaspi caerulescens, Iberis intermedia and Silene latifolia. This latter species was highlighted as a new thallium hyperaccumulator, accumulating nearly 1,500 mg kg^?C1. These species represent a patrimonial interest for their potential use for the phytoremediation of toxic metal-polluted areas.