Effects Of Drugs In Humans Research Articles

Morphological and functional characterization of human induced pluripotent stem cell-derived neurons (iCell Neurons) in defined culture systems.

Pre-clinical testing of drug candidates in animal models is expensive, time-consuming, and often fails to predict drug effects in humans. Industry and academia alike are working to build human-based in vitro test beds and advanced high throughput screening systems to improve the translation of preclinical results to human drug trials. Human neurons derived from induced pluripotent stems cells (hiPSCs) are readily available for use within these test-beds and high throughput screens, but there remains a need to robustly evaluate cellular behavior prior to their incorporation in such systems. This study reports on the characterization of one source of commercially available hiPSC-derived neurons, iCell(®) Neurons, for their long-term viability and functional performance to assess their suitability for integration within advanced in vitro platforms. The purity, morphology, survival, identity, and functional maturation of the cells utilizing different culture substrates and medium combinations were evaluated over 28 days in vitro (DIV). Patch-clamp electrophysiological data demonstrated increased capacity for repetitive firing of action potentials across all culture conditions. Significant differences in cellular maturity, morphology, and functional performance were observed in the different conditions, highlighting the importance of evaluating different surface types and growth medium compositions for application in specific in vitro protocols.

Demarcation of secondary hyperalgesia zones: Punctate stimulation pressure matters

BackgroundSecondary hyperalgesia is increased sensitivity in normal tissue near an injury, and it is a measure of central sensitization reflecting injury-related effects on the CNS. Secondary hyperalgesia areas (SHAs), usually assessed by polyamide monofilaments, are important outcomes in studies of analgesic drug effects in humans. However, since the methods applied in demarcating the secondary hyperalgesia zone seem inconsistent across studies, we examined the effect of a standardized approach upon the measurement of SHA following a first degree burn injury (BI). New methodThe study was a two-observer, test–retest study with the two sessions separated by 6wk. An observer-blinded design adjusted to examine day-to-day and observer-to-observer variability in SHA was used. In 23 healthy volunteers (12 females/11 males) a BI was induced by a contact thermode (47.0°C, 420s, 2.5×5.0cm2). The SHA, demarcated by polyamide monofilaments (bending force: 0.2, 69 and 2569mN) and a “weighted-pin” stimulator (512mN), were assessed 45 to 75min after each BI. ResultsA random effect, linear mixed model demonstrated a logarithmic correlation between elicited skin pressures (mN/mm2) and the SHAs (P<0.0001). No day-to-day or observer-to-observer differences in SHAs were observed. Intraclass correlation coefficients, in the range of 0.51 to 0.84, indicated a moderate to almost perfect reliability between observers. Comparison with existing methodsNo standardized approach in SHA-assessment has hitherto been presented. ConclusionsThis is the first study to demonstrate that demarcation of secondary hyperalgesia zones depends on the developed pressure of the punctate stimulator used.

Preclinical research: Make mouse studies work

More investment to characterize animal models can boost the ability of preclinical work to predict drug effects in humans, says Steve Perrin.

Discriminative stimulus effects of nitrous oxide in mice

The abuse-related behavioral effects produced by nitrous oxide (N₂O) gas have been suggested as being unique compared with other abused inhalants. The drug discrimination paradigm in animals can be used to study subjective effects of drugs in humans and to test this hypothesis. The goals of the present experiment were to establish N₂O discrimination in mice and to compare its discriminative stimulus effects with those of abused volatile vapors and vapor anesthetics. Sixteen B6SJLF1/J mice were trained to discriminate between 10 min of exposure to 60% N₂O+40% oxygen (O₂) and 10 min of exposure to 100% O₂. The time course of N₂O discrimination was examined, followed by cross-substitution testing with abused vapors, volatile anesthetics, ethanol, D-amphetamine, and 2-butanol. Mice acquired the ability to discriminate between N₂O and O₂ in 40 days. N₂O fully substituted for 10 min of exposure to 60% N₂O in a concentration-dependent manner. Full substitution required 7 min of 60% N₂O exposure, but the offset of stimulus effects following the cessation of exposure was more rapid. The aromatic hydrocarbon toluene almost fully substituted for N₂O. 1,1,1-Trichloroethane, methoxyflurane, isoflurane, and ethanol showed lesser degrees of substitution. D-amphetamine and the odorant 2-butanol did not substitute for N₂O. Given the varying degrees of incomplete substitution by test compounds, the discriminative stimulus properties of N₂O and, perhaps, its subjective effects in humans are probably not unique. As none of the inhalants tested fully mimicked N₂O, its overall effects may include one or more novel stimulus components.

Effects of psychotropic drugs on brain plasticity in humans

Although neurotransmitter-based hypotheses still prevail current thinking about the mechanism of action of psychotropic drugs, recent insight into the pathophysiology of psychiatric disorders has unveiled a range of new therapeutic actions of the drugs used to treat those disorders. Especially antidepressants seem to exert at least some of their effects via restoration of synaptic/neuronal plasticity. In addition, there is increasing evidence that several of the second-generation antipsychotics and some anticonvulsants affect neuronal survival/apoptosis as well as synaptic plasticity. Most of this evidence stems from work in animals. In this review, we will focus on the evidence for neuroplastic effects of psychotropic drugs in humans being aware of the fact that most of the data are derived from animals and that volumetric studies in humans can only indicate structural plasticity and not necessarily functional plasticity. However, as the data from human studies are rather poor and inconclusive, and sometimes even conflicting, it seems impossible to draw general conclusions. Until now studies on neuroplasticity in humans can only explain small pieces of the effects of psychotropic drugs on brain plasticity in humans. Nevertheless, future prospects for the development of new drugs targeting brain plasticity will be of importance and will complete this overview.

Relationship between drug discrimination and ratings of subjective effects

The discriminative and subjective effects of drugs in humans are related, but the full extent of this relationship remains to be determined. To further explore this relationship, a retrospective analysis was conducted on data from six studies completed in our laboratory that used identical procedures. The relationship between the discriminative and subjective effects of a range of doses of D-amphetamine (i.e. 2.5-15 mg) was examined using correlational analyses. Significant correlations with discrimination performance were observed on 15 of 20 items from the Drug-Effect Questionnaire across a range of qualities [e.g. Pay For (a positive effect indicative of abuse potential) and Active (a stimulant-like effect)], but the magnitude of these relationships was modest (r<0.52). The current findings demonstrate that diverse subjective effects contribute to the discriminative effects of D-amphetamine and indicate that the former are a more practical means to assess the abuse potential of drugs. Although these procedures are fundamentally related in that they rely on the presence of an interoceptive drug state, they differ in the dimension(s) of the interoceptive effects that participants must quantify. The simultaneous use of drug discrimination and subjective effects may, therefore, reveal complimentary aspects of drug effects that underlie their potential for abuse.

Sorafenib plus dacarbazine in solid tumors: a phase I study with dynamic contrast-enhanced ultrasonography and genomic analysis of sequential tumor biopsy samples

Improved prognostic accuracy for treatment response and a wider understanding of drug effects in humans are crucial for enhancing the utility of sorafenib and other promising targeted therapies. We developed a strategy of global genomic investigation of sequential tumor biopsy samples at baseline and 21 days post treatment, and applied this approach in a phase I study of sorafenib plus dacarbazine in patients with solid tumors. The objective of this study was also to validate functional parameters of DCE-US as surrogate markers to predict earlier response. Patients received 21-day cycles of oral sorafenib, 400 mg twice daily and dacarbazine, 1,000 mg/m(2) in a 1-h intravenous infusion on day 1. Efficacy was assessed using response evaluation criteria in solid tumors. Sequential biopsy samples (baseline and day 21) were obtained from the same tumor. Changes from baseline in global gene expression (GE) measured by genomic microarrays and in tumor vascularity at baseline, D8, D21, D 42 and every 2 cycles using dynamic contrast-enhanced ultrasonography (DCE-US) were analyzed for patients with and without a clinical response to treatment at 3 months. Among 23 patients evaluable for treatment efficacy, 17 were eligible for gene expression and DCE-US analyses. One patient achieved a partial response; 14 exhibited stable disease. Ten patients were defined as exhibiting stable disease (SD) and 7, progressive disease (PD) at 3 months. Genomic analyses identified a 237-gene signature that distinguished SD from PD at 3 months. Of note, CDK4 overexpression and PDGFR downregulation were associated with PD. Functional parameters of DCE-US representing the blood volume at baseline, day 8, and day 21 were correlated with disease progression at 3 months. This novel approach of sequential investigations in a phase I trial was feasible, detecting early changes in gene expression and tumor vascularity evaluated using DCE-US that may be predictive of clinical outcome.

HESI Cardic Safety Committee: Prospective studies to evaluate the sensitivity of animal models to predict effects of human drugs on cardiac contractility

HESI Cardic Safety Committee: Prospective studies to evaluate the sensitivity of animal models to predict effects of human drugs on cardiac contractility

Inattention, impulsive action, and subjective response to d-amphetamine

BackgroundBoth impulsivity and sensitivity to the rewarding effects of drugs have long been considered risk factors for drug abuse. There is some preclinical evidence to suggest that the two are related; however, there is little information about how specific behavioral components of impulsivity are related to the acute euphorigenic effects of drugs in humans. The aim of the current study was to examine the degree to which both inattention and impulsive action predicted subjective response to amphetamine. MethodsHealthy adults (n=165) performed the behavioral tasks and rated their subjective response to amphetamine (0, 5, 10, and 20mg). Inattention was assessed as attention lapses on a simple reaction time task, and impulsive action was measured by stop RT on the stop task. Subjective response to amphetamine was assessed with the Drug Effects Questionnaire (DEQ) and the Profile of Mood States (POMS). ResultsHierarchical linear regression analyses showed significant negative associations between attention lapses and subjective response to amphetamine on DEQ measures. By contrast, stop RT was positively associated with responses on both DEQ and POMS measures. Additionally, a dose-response relationship was observed, such that the strength of these associations increased with higher doses of amphetamine. ConclusionsThese findings suggest that inattention is associated with less subjective response to amphetamine. By contrast, the heightened sensitivity to stimulant drug reward observed in individuals high in impulsive action suggests that this might be one mechanism contributing to increased risk for stimulant drug abuse in these individuals.

Human brain functional network changes associated with enhanced and impaired attentional task performance.

How is the cognitive performance of the human brain related to its topological and spatial organization as a complex network embedded in anatomical space? To address this question, we used nicotine replacement and duration of attentionally demanding task performance (time-on-task), as experimental factors expected, respectively, to enhance and impair cognitive function. We measured resting-state fMRI data, performance and brain activation on a go/no-go task demanding sustained attention, and subjective fatigue in n = 18 healthy, briefly abstinent, cigarette smokers scanned repeatedly in a placebo-controlled, crossover design. We tested the main effects of drug (placebo vs Nicorette gum) and time-on-task on behavioral performance and brain functional network metrics measured in binary graphs of 477 regional nodes (efficiency, measure of integrative topology; clustering, a measure of segregated topology; and the Euclidean physical distance between connected nodes, a proxy marker of wiring cost). Nicotine enhanced attentional task performance behaviorally and increased efficiency, decreased clustering, and increased connection distance of brain networks. Greater behavioral benefits of nicotine were correlated with stronger drug effects on integrative and distributed network configuration and with greater frequency of cigarette smoking. Greater time-on-task had opposite effects: it impaired attentional accuracy, decreased efficiency, increased clustering, and decreased connection distance of networks. These results are consistent with hypothetical predictions that superior cognitive performance should be supported by more efficient, integrated (high capacity) brain network topology at greater connection distance (high cost). They also demonstrate that brain network analysis can provide novel and theoretically principled pharmacodynamic biomarkers of pro-cognitive drug effects in humans.

Recycling side-effects into clinical markers for drug repositioning

Side-effects are the unintended consequence of therapeutic treatments, but they can also be seen as valuable read-outs of drug effects in humans; these effects are difficult to infer or predict from pre-clinical models. Indeed, some studies suggest that drugs with similar side-effect profiles may also share therapeutic properties through related mechanisms of action. A recent publication exploits this concept to systematically investigate new indications for already marketed drugs, and presents a strategy to get the most out of the tiny portion of chemicals that have proved to be effective and safe.

Pharmacoproteomics Applications for Drug Target Discovery in CNS Disorders

Although target-based approach has yielded novel drugs in many therapeutic fields, this approach remains suboptimal and slows the pace of innovations for drug discovery in central nervous system (CNS) disorders. This is because the target identification requires a hierarchical integration from in vitro cellular and functional tissue studies to animal models that reasonably predict drug effects in humans. Because the majority of CNS drugs were discovered empirically to date, the precise target site(s) for many of them remain unknown or uncertain, even after years of clinical and commercial use. This paper presents a critical review and synthesis of the emerging proteomics applications for drug target discovery with an emphasis on defining the early molecular mechanisms by which (hitherto empirically discovered) drugs or novel neurotrophic factors initiate cellular change toward physiological effects with therapeutic ramifications. Pharmacoproteomics methodology provides important qualitative information on post-translational modifications to each protein as well as quantitative data on protein expression patterns in response to a given pharmaceutical stimulus. This information is particularly important because it provides data on early cellular events such as the stimulus and signaling cascades triggered independently of protein neosynthesis. Pharmacoproteomics is still in its infancy but offers substantial promise in personalized medicine in addition to the pharmacogenomics methodologies. Keywords: CNS, diagnostic medicine, drug targets, personalized medicine, pharmacoproteomics, Disorders, novel neurotrophic factors, Alzheimer's Disease, Parkinson's Disease, Depression, Schizophrenia, Bipolar Disorder

Comparison of the novel HK-2 human renal proximal tubular cell line with the standard LLC-PK1 cell line in studying drug-induced nephrotoxicity

Established cell lines are widely used as in vitro models in toxicology studies. The choice of an appropriate cell line is critical when performing studies to elucidate drug-induced toxicity in humans. The porcine renal proximal tubular cell line LLC-PK1 is routinely used to study the nephrotoxic effects of drugs in humans. However, there are significant interspecies differences in drug pharmacokinetics and pharmacodynamics. The objective of this study was to determine whether the human renal proximal tubular cell line HK-2 is an acceptable model to use when performing in vitro toxicity studies to predict effects in humans. We examined 2 nephrotoxic agents, ifosfamide (IFO) and acyclovir, that exhibit different clinical nephrotoxic patterns. HK-2 cells metabolized IFO to its nephrotoxic metabolite, chloroacetaldehyde (CAA). Acyclovir induced a concentration-dependent decrease in HK-2 cell viability, suggesting that acyclovir may induce direct insult to renal proximal tubular cells. The results support clinical pathology data in humans and suggest that HK-2 cells are a suitable model to use in in vitro toxicity studies to determine drug-induced nephrotoxicity in humans.

REVIEW: Reward sensitivity: issues of measurement, and achieving consilience between human and animal phenotypes

Reward is a concept fundamental to discussions of drug abuse and addiction. The idea that altered sensitivity to either drug-reward, or to rewards in general, contributes to, or results from, drug-taking is a common theme in several theories of addiction. However, the concept of reward is problematic in that it is used to refer to apparently different behavioural phenomena, and even to diverse neurobiological processes (reward pathways). Whether these different phenomena are different behavioural expressions of a common underlying process is not established, and much research suggests that there may be only loose relationships among different aspects of reward. Measures of rewarding effects of drugs in humans often depend upon subjective reports. In animal studies, such insights are not available, and behavioural measures must be relied upon to infer rewarding effects of drugs or other events. In such animal studies, but also in many human methods established to objectify measures of reward, many other factors contribute to the behaviour being studied. For that reason, studying the biological (including genetic) bases of performance of tasks that ostensibly measure reward cannot provide unequivocal answers. The current overview outlines the strengths and weaknesses of current approaches that hinder the conciliation of cross-species studies of the genetics of reward sensitivity and the dysregulation of reward processes by drugs of abuse. Some suggestions are made as to how human and animal studies may be made to address more closely homologous behaviours, even if those processes are only partly able to isolate 'reward' from other factors contributing to behavioural output.

The butyrylcholinesterase knockout mouse a research tool in the study of drug sensitivity, bio-distribution, obesity and Alzheimer's disease

Butyrylcholinesterase (BChE) mutations common in the human population may result in complete or partial BChE deficiency, making the BChE knockout (KO) mouse a model for human deficiencies. The BChE KO mouse cannot tolerate standard doses of the muscle relaxant succinylcholine or the Alzheimer's disease drugs huperzine A and donepezil. It is resistant to the asthma drug bambuterol. The importance of BChE in detoxication of cocaine has been demonstrated by hepatotoxicity and cardiotoxicity in cocaine-challenged BChE KO mice. The BChE KO mouse becomes obese on a high-fat diet, suggesting a role for BChE in fat metabolism. BChE serves as a backup for acetylcholinesterase by hydrolyzing the neurotransmitter acetylcholine in acetylcholinesterase knockout mice. Imaging studies show that BChE injected intrathecally crosses the blood–brain barrier. Mice, but not humans, have carboxylesterase in their blood. Carboxylesterase obscures the role of BChE in detoxication of organophosphorus pesticides. Future studies will make a double knockout that has neither BChE nor carboxylesterase. The double knockout is expected to be unusually sensitive to the toxicity of organophosphorus pesticides. Knowledge of drug sensitivities in the mouse model of human BChE deficiency will aid in understanding adverse drug effects in humans.

Characterization of an inhaled toluene drug discrimination in mice: Effect of exposure conditions and route of administration

The drug discrimination procedure in animals has been extensively utilized to model the abuse related, subjective effects of drugs in humans, but it has seldom been used to examine abused volatile inhalants like toluene. The present study sought to characterize the temporal aspects of toluene's discriminative stimulus as well assess toluene blood concentrations under identical exposure conditions. B6SJLF1/J mice were trained to discriminate 10 min of exposure to 6000 ppm inhaled toluene vapor from air. Toluene vapor concentration dependently substituted for the training exposure condition with longer exposures to equivalent concentrations producing greater substitution than shorter exposures. Toluene's discriminative stimulus effects dissipated completely by 60 min after the cessation of exposure. Injected liquid toluene dose-dependently substituted for toluene vapor as well as augmenting the discriminative stimulus effects of inhaled toluene. Toluene blood concentrations measured under several exposure conditions which produced full substitution were all nearly identical suggesting that the concentration of toluene in the animal tissues at the time of testing determined discriminative performance. These results indicate that the discriminative stimulus effects of inhaled toluene vapor are likely mediated by CNS effects rather than by its pronounced peripheral stimulus effects.

The phencyclidine-like discriminative stimulus effects and reinforcing properties of the NR2B-selective N-methyl-D-aspartate antagonist CP-101 606 in rats and rhesus monkeys

Development of N-methyl-D-aspartate (NMDA) antagonists for a variety of disorders has been hindered by their production of phencyclidine (PCP)-like psychological effects and abuse potential. There is, however, evidence to suggest that this problem might be mitigated by targeting NMDA receptors subtypes, in particular, those containing the NR2B subunit. To further test this hypothesis, the NR2B selective antagonist CP-101 606 (traxoprodil) was evaluated in two animal models: drug discrimination, a model of the subjective effects of drugs in humans, and self-administration, which evaluates the reinforcing properties of the drug. In the first study, CP-101 606(3-300 microg/kg/infusion) was tested for intravenous self-administration in rhesus monkeys experienced in PCP (5.6 microg/kg/infusion, intravenously) self-administration. In the second study, CP-101 606 was tested for production of PCP-like discriminative stimulus effects in rats (3-56 mg/kg, intraperitoneally) and rhesus monkeys (0.3-5.6 mg/kg intravenously). Evidence was obtained for reinforcing effects of at least one dose of CP-101 606 in all four monkeys. In rats, CP-101 606 produced more than 80% mean PCP-lever selection (2.0 mg/kg, intraperitoneally) but, unlike PCP itself, the dose producing the highest level of substitution was accompanied by more than 50% suppression of response rates. In monkeys, CP-101 606 produced more than 90% PCP-lever selection (0.1 mg/kg intramuscularly) in three of four animals at doses that did not significantly decrease rates of responding. The data show that CP-101 606 has some PCP-like discriminative stimulus effects in rats and monkeys and functions as a positive reinforcer in monkeys. These results suggest that inhibition of NR2B subunit containing NMDA receptors plays a role in the production of the subjective effects and abuse potential associated with many subtype-nonselective NMDA receptor antagonists such as PCP.

A New Look at the Profession of Clinical Pharmacology

The debate over the scope of clinical pharmacology--and the clinical pharmacologist--is ongoing. The stated content of clinical pharmacology is therapeutics, whereas the recommended training programs are geared to train researchers. Most clinical pharmacologists make their living in research. People who do not identify themselves as clinical pharmacologists do much research on drug effects in humans. The professional clinical pharmacologist's role must be re-evaluated in the context of society's needs and sources of funding.

Thinking inside the box

In the next few decades, brain medicine will present a particular socioeconomic challenge for ageing citizens worldwide. Many disorders and ailments that affect the brain—including Alzheimer disease, Parkinson disease, dementia and stroke—are chronic conditions that persist for years or even decades. In addition, many of these disorders have devastating effects, which together create a substantial burden on society: about one‐third of the global disease burden can be attributed to disorders of the brain or nervous system (Olesen & Leonardi, 2003; Andlin‐Sobocki et al , 2005). In the USA alone, there are as many as 5.5 million individuals with Alzheimer disease, 1.5 million with Parkinson disease and 400,000 with multiple sclerosis. These devastating diseases not only place a heavy emotional burden on patients and their care‐givers, but also have an important economic impact. > …about one‐third of the global disease burden can be attributed to disorders of the brain or nervous system Unfortunately, this tremendous socioeconomic problem is worsening, because the risk of suffering from a brain disease increases with age, and life expectancy is increasing worldwide, particularly in developed countries. In the case of Alzheimer disease, for example, ∼10% of US citizens aged over 65 years are affected, the prevalence rate has more than doubled since 1980 and it could increase twofold to threefold by 2050 if the current trend continues (Alzheimer's Association, 2006). Unfortunately, research has had little impact on these statistics. Despite vigorous efforts from both the pharmaceutical industry and biomedical researchers, there are still no disease‐modifying treatments available for Alzheimer disease, multiple sclerosis, stroke and a range of other neurological disorders. Some have argued that this problem cannot be solved by standard high‐throughput target‐based drug discovery methods, because we simply do not know which mechanisms to target. Instead, we should revert to previous empirical models of drug …

Noninvasive brain imaging for experimental medicine in drug discovery

There is a widely shared perception that it is becoming increasingly urgent to find reliable measures related to therapeutic efficacy for use as early as possible in drug development. The inadequate understanding of diseases, limitations of animal models and difficulties in using their responses to anticipate drug effects in humans highlights the need to develop tools in experimental medicine to characterise human disease directly. Noninvasive imaging, particularly positron emission tomography and magnetic resonance imaging, provide a powerful range of methods for serial observation of drug distribution and interactions, and for assessing potential therapeutic mechanisms. Using imaging technology to establish biological proof-of-principle and as a pharmacodynamic marker for dose ranging would contribute greatly to the speed and efficiency of early decision making in new drug development. Imaging methods offer the ultimate promise for the development of clinically predictive surrogate markers of disease responses to new drugs.