Vasoactivity of AG014699, a clinically active small molecule inhibitor of poly(ADP-ribose) polymerase: a contributory factor to chemopotentiation in vivo?

Purpose: Poly(ADP-ribose) polymerase (PARP) plays an important role in DNA repair, and PARP inhibitors can enhance the activity of DNA-damaging agents in vitro and in vivo. AG014699 is a potent PARP inhibitor in phase II clinical development. However, the range of therapeutics with which AG014699 could interact via a DNA-repair based mechanism is limited. We aimed to investigate a novel, vascular-based activity of AG014699, underlying in vivo chemosensitization, which could widen its clinical application.

Experimental Design: Temozolomide response was analyzed in vitro and in vivo. Vessel dynamics were monitored using “mismatch” following the administration of perfusion markers and real-time analysis of fluorescently labeled albumin uptake in to tumors established in dorsal window chambers. Further mechanistic investigations used ex vivo assays of vascular smooth muscle relaxation, gut motility, and myosin light chain kinase (MLCK) inhibition.

Results: AG014699 failed to sensitize SW620 cells to temozolomide in vitro but induced pronounced enhancement in vivo. AG014699 (1 mg/kg) improved tumor perfusion comparably with the control agents nicotinamide (1 g/kg) and AG14361 (forerunner to AG014699; 10 mg/kg). AG014699 and AG14361 relaxed preconstricted vascular smooth muscle more potently than the standard agent, hydralazine, with no impact on gut motility. AG014699 inhibited MLCK at concentrations that relaxed isolated arteries, whereas AG14361 had no effect.

Conclusion: Increased vessel perfusion elicited by AG014699 could increase tumor drug accumulation and therapeutic response. Vasoactive concentrations of AG014699 do not cause detrimental side effects to gut motility and may increase the range of therapeutics with which AG014699 could be combined with for clinical benefit.

Inhibitor profiling of the Pseudomonas aeruginosa virulence factor LasB using N-alpha mercaptoamide template-based inhibitors

We report on the synthesis and biological evaluation of a focussed library of N-alpha mercaptoamide containing dipeptides as inhibitors of the zinc metallopeptidase Pseudomonas aeruginosa elastase (LasB, EC 3.4.24.26). The aim of the study was to derive an inhibitor profile for LasB with regard to mapping the S´1 binding site of the enzyme. Consequently, a focussed library of 160 members has been synthesised, using standard Fmoc-solid phase methods (on a Rink-amide resin), in which a subset of amino acids including examples of those with basic (Lys, Arg), aromatic (Phe, Trp), large aliphatic (Val, Leu) and acidic (Asp, Glu) side-chains populated the P´2 position of the inhibitor sequence and all 20 natural amino acids were incorporated, in turn, at the P´1 position. The study has revealed a preference for aromatic and/or large aliphatic amino acids at P´1 and a distinct bias against acidic residues at P´2. Ten inhibitor sequences were discovered that exhibited sub to low micromolar Ki values.

Quality factor assessment of subwavelength resonant cavities at FIR frequencies

Subwavelength resonators at FIR are presented and studied. The structures consist of 1D cavities formed between a metallized (silver) surface and a metamaterial surface comprising a periodic array of silver patches on a silver-backed silicon substrate. The concept derives from recent discoveries of artificial magnetic conductors (AMC). By studying the currents excited on the metamaterial surface by a normally incident plane wave, the nature of the emerging resonant phenomena and the physical mechanism underlying the AMC operation are investigated. Full wave simulations, based on finite element method and time-domain transmission line modelling technique, have been carried out to demonstrate the effective AMC boundary condition and prove the possibilities for subwavelength cavities. The quality factor of the resonant cavities is assessed as a function of the cavity profile. It is demonstrated that the quality factor drops to about 1/8 of the half-wavelength value for lambda/8 resonant cavity.

Confirmatory factor analysis of the adolescent self-report strengths and difficulties questionnaire

The Strengths and Difficulties Questionnaire (SDQ) is a widely used 25-item screening test for emotional and behavioral problems in children and adolescents. This study attempted to critically examine the factor structure of the adolescent self-report version. As part of an ongoing longitudinal cohort study, a total of 3,753 pupils completed the SDQ when aged 12. Both three- and five-factor exploratory factor analysis models were estimated. A number of deviations from the hypothesized SDQ structure were observed, including a lack of unidimensionality within particular subscales, cross-loadings, and items failing to load on any factor. Model fit of the confirmatory factor analysis model was modest, providing limited support for the hypothesized five-component structure. The analyses suggested a number of weaknesses within the component structure of the self-report SDQ, particularly in relation to the reverse-coded items.