Unexpected neurotoxicity in chronic toxicity studies with a HBV viral expression inhibitor.

The non-clinical safety profile of the small molecule hepatitis B virus viral expression inhibitor RG7834 was studied in a package consisting of safety pharmacology, genotoxicity, repeat dose toxicity and reproductive toxicity studies. The chronic monkey toxicity study identified dose- and time-dependent symptoms of polyneuropathy, with correlating nerve conduction velocity reductions and axonal degeneration in peripheral nerves and spinal cord, in all compound treatment groups with no evidence of reversibility after approximately 3 months of treatment cessation. Similar histopathological findings were observed in the chronic rat toxicity study. Subsequent in vitro neurotoxicity investigations and ion channel electrophysiology did not elucidate a potential mechanism for the late toxicity. However, based on similar findings observed with a structurally different molecule, an inhibition of their common pharmacological targets, PAPD5 & PAPD 7, was considered as a possible mechanism of toxicity. In conclusion, the marked neuropathies, only observed after chronic dosing, did not support further clinical development of RG7834 because of its foreseen clinical treatment duration of up to 48 weeks in chronic HBV patients.

Characterization of 2-[[4-fluoro-3-(trifluoromethyl)phenyl]amino]-4-(4-pyridinyl)-5-thiazolemethanol (JNJ-1930942), a novel positive allosteric modulator of the {alpha}7 nicotinic acetylcholine receptor.

The α(7) nicotinic acetylcholine receptor (nAChR) is a potential therapeutic target for the treatment of cognitive deficits associated with schizophrenia, Alzheimer's disease, Parkinson's disease, and attention-deficit/hyperactivity disorder. Activation of α(7) nAChRs improved sensory gating and cognitive function in animal models and in early clinical trials. Here we describe the novel highly selective α(7) nAChR positive allosteric modulator, 2-[[4-fluoro-3-(trifluoromethyl)phenyl]amino]-4-(4-pyridinyl)-5-thiazolemethanol (JNJ-1930942). This compound enhances the choline-evoked rise in intracellular Ca(2+) levels in the GH4C1 cell line expressing the cloned human α(7) nAChR. JNJ-1930942 does not act on α4ß2, α3ß4 nAChRs or on the related 5-HT3A channel. Electrophysiological assessment in the GH4C1 cell line shows that JNJ-1930942 increases the peak and net charge response to choline, acetylcholine, and N-[(3R)-1-azabicyclo[2.2.2]oct-3-yl]-4-chlorobenzamide (PNU-282987). The potentiation is obtained mainly by affecting the receptor desensitization characteristics, leaving activation and deactivation kinetics as well as recovery from desensitization relatively unchanged. Choline efficacy is increased over its full concentration response range, and choline potency is increased more than 10-fold. The potentiating effect is α(7) channel-dependent, because it is blocked by the α(7) antagonist methyllycaconitine. Moreover, in hippocampal slices, JNJ-1930942 enhances neurotransmission at hippocampal dentate gyrus synapses and facilitates the induction of long-term potentiation of electrically evoked synaptic responses in the dentate gyrus. In vivo, JNJ-1930942 reverses a genetically based auditory gating deficit in DBA/2 mice. JNJ-1930942 will be a useful tool to study the therapeutic potential of α(7) nAChR potentiation in central nervous system disorders in which a deficit in α(7) nAChR neurotransmission is hypothesized to be involved.

Reprint of: Nonclinical species sensitivity to convulsions: An IQ DruSafe consortium working group initiative.

Clinical development of compounds that carry a convulsion liability is typically limited by safety margins based on the most sensitive nonclinical species. To better understand differences in sensitivity to drug-induced convulsion of commonly used preclinical species, a survey was distributed amongst pharmaceutical companies through an IQ consortium (International Consortium for Innovation and Quality in Pharmaceutical Development) resulting in convulsion-related data on 80 unique compounds from 11 companies. The lowest free drug plasma concentration at which convulsions were observed and the no observed effect level for convulsions were compared between species to determine their relative sensitivity. Additionally, data were collected on other endpoints including use of electroencephalography, premonitory signs, convulsion type, the reason why development was stopped, and the highest development phase reached. The key outcomes were: (1) the dog was most often determined to be the most sensitive species by both non-exposure and exposure-based analyses, (2) there was not a clear sensitivity ranking of other species (NHP, rat and mouse), (3) CNS symptoms were frequently present at exposures that were not associated with convulsions, but no single reliable premonitory indicator of convulsion was identified, and (4) the lack of convulsions in the compounds that were tested in humans in this dataset may suggest that convulsion liability is well mitigated via current drug development strategies.

Nonclinical species sensitivity to convulsions: An IQ DruSafe consortium working group initiative.

Clinical development of compounds that carry a convulsion liability is typically limited by safety margins based on the most sensitive nonclinical species. To better understand differences in sensitivity to drug-induced convulsion of commonly used nonclinical species, a survey was distributed amongst pharmaceutical companies through an IQ consortium (International Consortium for Innovation and Quality in Pharmaceutical Development) resulting in convulsion-related data on 80 unique compounds from 11 companies. The lowest free drug plasma concentration at which convulsions were observed and the no observed effect level for convulsions were compared between species to determine their relative sensitivity. Additionally, data were collected on other endpoints including use of electroencephalography, premonitory signs, convulsion type, the reason why development was stopped, and the highest development phase reached. The key outcomes were: (1) the dog was most often determined to be the most sensitive species by both non-exposure and exposure-based analyses, (2) there was not a clear sensitivity ranking of other species (NHP, rat and mouse), (3) CNS symptoms were frequently present at exposures that were not associated with convulsions, but no single reliable premonitory indicator of convulsion was identified, and (4) the lack of convulsions when compounds were tested in humans in this dataset may suggest that convulsion liability is well mitigated via current drug development strategies.

A Framework Proposal to Follow-Up on Preclinical Convulsive Signals of a New Molecular Entity in First-in-Human Studies Using Electroencephalographic Monitoring.

Traditionally, in dose-escalating first-in-human (FiH) studies, a dose cap with a 10-fold safety margin to the no observed effect level in animals is implemented if convulsive events are observed in animals. However, the convulsive risk seen in animals does not generally translate to humans. Several lines of evidence are summarized indicating that in a dose-escalating setting, electroencephalographic epileptiform abnormalities occur at lower doses than clinical convulsive events. Therefore, we propose to consider the occurrence of epileptiform abnormalities in toxicology studies as premonitory signals for convulsions in dose-escalating FiH studies. Compared with the traditional dose-cap approach, this may allow the exploration of higher doses in FiH and, subsequently, phase II studies without compromising human safety. Similarly, the presence or absence of electroencephalographic epileptiform abnormalities may also aid the assessment of proconvulsive risk in situations of increased perpetrator burden as potentially present in pharmacokinetic and/or pharmacodynamic drug-drug interactions.

Frequency-dependent properties of a fluid jet stimulus: calibration, modeling, and application to cochlear hair cell bundles.

The investigation of small physiological mechano-sensory systems, such as hair cells or their accessory structures in the inner ear or lateral line organ, requires mechanical stimulus equipment that allows spatial manipulation with micrometer precision and stimulation with amplitudes down to the nanometer scale. Here, we describe the calibration of a microfluid jet produced by a device that was designed to excite individual cochlear hair cell bundles or cupulae of the fish superficial lateral line system. The calibration involves a precise definition of the linearity and time- and frequency-dependent characteristics of the fluid jet as produced by a pressurized fluid-filled container combined with a glass pipette having a microscopically sized tip acting as an orifice. A procedure is described that can be applied during experiments to obtain a fluid jet's frequency response, which may vary with each individual glass pipette. At small orifice diameters (<15 mum), the fluid velocity of the jet is proportional to the displacement of the piezoelectric actuator pressurizing the container's volume and is suitable to stimulate the hair bundles of sensory hair cells. With increasing diameter, the fluid jet velocity becomes proportional to the actuator's velocity. The experimentally observed characteristics can be described adequately by a dynamical model of damped fluid masses coupled by elastic components.

Desensitization characteristics of the human alpha7nAChR/5HT3A chimera receptor.

The alpha7-nicotinic acetylcholine receptor (alpha7) is an important ionotropic receptor in the central nervous system, which becomes permeable to cations upon binding of its natural agonist acetylcholine (ACh). alpha7 kinetics are characterized by rapid activation, followed by fast desensitization of the current. As the wild-type (WT) alpha7 is difficult to express heterologously in mammalian cellular systems, frequently a more easily expressible chimera consisting of the extracellular domain of the alpha7 and the transmembrane domain of the 5HT3A receptor is used to study alpha7 pharmacology (chick alpha7/mouse 5HT3A [Eiselé et al., 1993]; human alpha7/mouse 5HT3A [Graig et al., 2004]). Desensitization characteristics of these chimera receptors have been described as intermediate compared with the fast desensitizing alpha7 and the more slowly desensitizing 5HT3A receptors. Here, we describe a fully human chimera receptor (h-alpha7/5HT3A), which is characterized by desensitization, and recovery kinetics that deviate from the human WT alpha7.

Channel gating forces govern accuracy of mechano-electrical transduction in hair cells.

Sensory hair cells are known for the exquisite displacement sensitivity with which they detect the sound-evoked vibrations in the inner ear. In this article, we determine a stochastically imposed fundamental lower bound on a hair cell's sensitivity to detect mechanically coded information arriving at its hair bundle. Based on measurements of transducer current and its noise in outer hair cells and the application of estimation theory, we show that a hair cell's transducer current carries information that allows the detection of vibrational amplitudes with an accuracy on the order of nanometers. We identify the transducer channel's molecular gating force as the physical factor controlling this accuracy in proportion to the inverse of its magnitude. Further, we show that the match of stochastic channel noise to gating-spring noise implies that the gating apparatus operates at the threshold of negative stiffness.