Preclinical to Clinical Translation of CNS Transporter Occupancy of TD-9855, a Novel Norepinephrine and Serotonin Reuptake Inhibitor

Background:

Monoamine reuptake inhibitors exhibit unique clinical profiles that reflect distinct engagement of the central nervous system (CNS) transporters.

Methods:

We used a translational strategy, including rodent pharmacokinetic/pharmacodynamic modeling and positron emission tomography (PET) imaging in humans, to establish the transporter profile of TD-9855, a novel norepinephrine and serotonin reuptake inhibitor.

Results:

TD-9855 was a potent inhibitor of norepinephrine (NE) and serotonin 5-HT uptake in vitro with an inhibitory selectivity of 4- to 10-fold for NE at human and rat transporters. TD-9855 engaged norepinephrine transporters (NET) and serotonin transporters (SERT) in rat spinal cord, with a plasma EC₅₀ of 11.7ng/mL and 50.8ng/mL, respectively, consistent with modest selectivity for NET in vivo.Accounting for species differences in protein binding, the projected human NET and SERT plasma EC₅₀ values were 5.5ng/mL and 23.9ng/mL, respectively. A single-dose, open-label PET study (4–20mg TD-9855, oral) was conducted in eight healthy males using the radiotracers [¹¹C]-3-amino-4- [2-[(di(methyl)amino)methyl]phenyl]sulfanylbenzonitrile for SERT and [¹¹C]-(S,S)-methylreboxetine for NET. The long pharmacokinetic half-life (30–40h) of TD-9855 allowed for sequential assessment of SERT and NET occupancy in the same subject. The plasma EC₅₀ for NET was estimated to be 1.21ng/mL, and at doses of greater than 4mg the projected steady-state NET occupancy is high (>75%). After a single oral dose of 20mg, SERT occupancy was 25 (±8)% at a plasma level of 6.35ng/mL.

Conclusions:

These data establish the CNS penetration and transporter profile of TD-9855 and inform the selection of potential doses for future clinical evaluation.     

Sublethal Effects of Waterborne Herbicides in Tropical Freshwater Fish

The study evaluated the sublethal effects of the herbicides glyphosate (Roundup) and diuron (Hexaron) and the mixture of them, used extremely in agriculture, through biomarkers in fish. The glutathione S-transferase activity increased (74%) and catalase activity decreased (37%) at the higher exposure concentration of Hexaron in comparison to the control group, suggesting an activation of this metabolism route. Membrane damage was observed at the higher exposure of Roundup and in the mixture group compared to the control group, which can be related to the nuclear alterations observed in these exposed groups. The cholinesterase activity was also inhibited (37%) in mixture group compared to the control group and no gill morphology damage was found. The results suggested a potential synergic effect in some analysed parameters.     

Error variance and sensitivity associated with three different Periotron® instruments

Calibration curves for three different Periotron® instruments, two model HAR-6000 and one HAR-600, were determined using human serum. These curves revealed a quadratic relationship between the volume and Periotron® readout for all three models. The calibration curve data were subjected to an analysis of covariance to test the equality of the three slopes. These were found to be different (p < 0.00001) and pairwise comparisons of the slopes were done. The procedure showed that the differences, as measured by the different slopes of the three instruments, were statistically significant (p < 0.0001). Although there were only small differences in the response to volume ratios, and hence also in the slopes of the calibration curves of the two HAR-6000 models, these were nevertheless significant (p ≤ 0.001). All three instruments showed good reproducibility as determined by error variances. The newer of the two HAR-6000 models showed the best reproducibility while the other had values similar to those of the HAR-600 instrument. It was concluded that the significantly differing response: volume ratios for the various Periotron® models necessitate the construction of calibration curves for these instruments before and during clinical studies. In addition, our data enable regular assessment of instrument reproducibility, which is an essential procedure in any long-term project using them to measure periodontal status.     

Progressive potentiation of peptide release during a neuronal discharge

1. In response to electrical stimulation, the bag cell neurons of Aplysia generate an afterdischarge that lasts 20-40 min. During this afterdischarge several neuroactive peptides are released. We have now studied the time course of release of two of these peptides, egg-laying hormone (ELH) and acidic peptide (AP). For the collection of released peptides, the artery to the bag cell clusters was perfused. The medium surrounding the clusters (artificial seawater, ASW) was completely exchanged at 5-min intervals before, during, and after stimulation of an afterdischarge. Peptides released into the external medium were analyzed with the use of high-pressure liquid chromatography. 2. Before stimulation, no detectable ELH and AP were found in the external medium. After the onset of an afterdischarge, the amount of ELH and AP released increased progressively until 15-20 min of firing. Toward the conclusion of an afterdischarge, the release of ELH and AP returned to control levels. 3. In contrast to the pattern of release of the peptides, the firing rate of the bag cell neurons is maximal within the first minute of afterdischarge and thereafter declines. 4. Release of the peptides from axonal varicosities occurs within the vascularized connective-tissue sheath that covers the clusters of bag cell neurons. Experiments were therefore carried out to establish whether the observed time course of release is affected by diffusion of the peptides through the vasculature into the external medium and, in particular, to determine whether the maximal rate of release at 15-20 min into the afterdischarge could be accounted for by a delay in transport of peptides from the neurites.(ABSTRACT TRUNCATED AT 250 WORDS)     

Screening of army soldiers for Wuchereria bancrofti infection in the metropolitan Recife region, Brazil: implications for epidemiological surveillance

Between 1989 and 1995, blood surveys were performed for Wuchereria bancrofti infection in several barracks of the Brazilian army in the metropolitan Recife region. For initial screening, 60 microliters of capillary blood were examined for microfilaria. All men who tested positive had microfilaria quantified by filtration of venous blood through a polycarbonate membrane. Of 23,773 men screened, 585 (2.5%) had microfilaria (mf). Microfilarial density ranged from < 1-8706 mf/ml of blood. Thirteen individuals had ultra-low microfilarial densities (1 mf/11 ml of blood). Characterization of 174 autochthonous cases made it possible to map 8 new districts in 4 cities within metropolitan Recife region where transmission of W. bancrofti was previously unknown. Routine screening of soldiers in the military may provide important surveillance data for national programmes to eliminate transmission of W. bancrofti.