Synthesis, Kinetic Evaluation and Utilization of a Biotinylated Dipeptide Proline Diphenyl Phosphonate for the Disclosure of Dipeptidyl Peptidase IV-like Serine Proteases

In this study, we report on the synthesis, kinetic characterisation, and application of a novel biotinylated and active site-directed inactivator of dipeptidyl peptidase IV (DPP-IV). Thus, the dipeptide-derived proline diphenyl phosphonate NH(2)-Glu(biotinyl-PEG)-Pro(P)(OPh)(2) has been prepared by a combination of classical solution- and solid-phase methodologies and has been shown to be an irreversible inhibitor of porcine DPP-IV, exhibiting an over all second-order rate constant (k(i)/K(i)) for inhibition of 1.57 x 10(3) M(-1) min(-1). This value compares favourably with previously reported rates of inactivation of DPP-IV by dipeptides containing a P(1) proline diphenyl phosphonate grouping [B. Boduszek, J. Oleksyszyn, C.M. Kam, J. Selzler, R.E. Smith, J.C. Powers, Dipeptide phophonates as inhibitors of dipeptidyl peptidase IV, J. Med. Chem. 37 (1994) 3969-3976; B.F. Gilmore, J.F. Lynas, C.J. Scott, C. McGoohan, L. Martin, B. Walker, Dipeptide proline diphenyl phosphonates are potent, irreversible inhibitors of seprase (FAPalpha), Biochem, Biophys. Res. Commun. 346 (2006) 436-446.], thus demonstrating that the incorporation of the side-chain modified (N-biotinyl-3-(2-(2-(3-aminopropyloxy)-ethoxy)-ethoxy)-propyl) glutamic acid residue at the P(2) position is compatible with inhibitor efficacy. The utilisation of this probe for the detection of both purified dipeptidyl peptidase IV and the disclosure of a dipeptidyl peptidase IV-like activity from a clinical isolate of Porphyromonas gingivalis, using established electrophoretic and Western blotting techniques previously developed by our group, is also demonstrated.

An orally available compound prevents deficits in memory caused by the Alzheimer amyloid-B oligomers.

Despite progress in defining a pathogenic role for amyloid beta protein (Abeta) in Alzheimer's disease, orally bioavailable compounds that prevent its effects on hippocampal synaptic plasticity and cognitive function have not yet emerged. A particularly attractive therapeutic strategy is to selectively neutralize small, soluble Abeta oligomers that have recently been shown to mediate synaptic dysfunction. METHODS: Using electrophysiological, biochemical, and behavioral assays, we studied how scyllo-inositol (AZD-103; molecular weight, 180) neutralizes the acutely toxic effects of Abeta on synaptic function and memory recall. RESULTS: Scyllo-inositol, but not its stereoisomer, chiro-inositol, dose-dependently rescued long-term potentiation in mouse hippocampus from the inhibitory effects of soluble oligomers of cell-derived human Abeta. Cerebroventricular injection into rats of the soluble Abeta oligomers interfered with learned performance on a complex lever-pressing task, but administration of scyllo-inositol via the drinking water fully prevented oligomer-induced errors. INTERPRETATION: A small, orally available natural product penetrates into the brain in vivo to rescue the memory impairment produced by soluble Abeta oligomers through a mechanism that restores hippocampal synaptic plasticity.

Electron impact excitation of C IV

Energy levels and radiative rates for transitions among the lowest 24 fine-structure levels belonging to the ls(2) nl (n <5) configurations of Li-like C IV have been calculated using the fully relativistic GRASP code. Additionally, collision strengths for transitions among these levels have been computed over a wide energy range below 28 Ry, using the Dirac Atomic R- matrix Code. Resonances have been resolved in a fine energy mesh in order to calculate the effective collision strengths. Results for radiative rates, collision strengths, and effective collision strengths are presented for all transitions. Comparisons with other available results are made, and the accuracy of the present data is assessed. Energy levels are expected to be accurate to within 1%, while other parameters are probably accurate to better than 20%.

Multi-zone TAP-reactors theory and application - IV. Ideal and non-ideal boundary conditions

We study the influence of non-ideal boundary and initial conditions (BIC) of a temporal analysis of products (TAP) reactor model on the data (observed exit flux) analysis. The general theory of multi-response state-defining experiments for a multi-zone TAP reactor is extended and applied to model several alternative boundary and initial conditions proposed in the literature. The method used is based on the Laplace transform and the transfer matrix formalism for multi-response experiments. Two non-idealities are studied: (1) the inlet pulse not being narrow enough (gas pulse not entering the reactor in Dirac delta function shape) and (2) the outlet non-ideality due to imperfect vacuum. The effect of these non-idealities is analyzed to the first and second order of approximation. The corresponding corrections were obtained and discussed in detail. It was found that they are negligible. Therefore, the model with ideal boundary conditions is proven to be completely adequate to the description and interpretation of transport-reaction data obtained with TAP-2 reactors.