Andrei L. Kleschyov

Number of nitrate groups determines reactivity and potency of organic nitrates: a proof of concept study in ALDH-2-/- mice.

Mitochondrial aldehyde dehydrogenase (ALDH‐2) has been shown to provide a pathway for bioactivation of organic nitrates and to be prone to desensitization in response to highly potent, but not to less potent, nitrates. We therefore sought to support the hypothesis that bioactivation by ALDH‐2 critically depends on the number of nitrate groups within the nitrovasodilator.

Sirolimus-Induced Vascular Dysfunction: Increased Mitochondrial and Nicotinamide Adenosine Dinucleotide Phosphate Oxidase-Dependent Superoxide Production and Decreased Vascular Nitric Oxide Formation

OBJECTIVES This study sought to analyze mechanisms that mediate vascular dysfunction induced by sirolimus. BACKGROUND Despite excellent antirestenotic capacity, sirolimus-eluting stents have been found to trigger coronary endothelial dysfunction and impaired re-endothelialization. METHODS To mimic the continuous sirolimus exposure of a stented vessel, Wistar rats underwent drug infusion with an osmotic pump for 7 days. RESULTS Sirolimus treatment caused a marked degree of endothelial dysfunction as well as a desensitization of the vasculature to the endothelium-independent vasodilator nitroglycerin. Also, sirolimus stimulated intense transmural superoxide formation as detected by dihydroethidine fluorescence in aortae. Increased superoxide production was mediated in part by the vascular nicotinamide adenosine dinucleotide phosphate (NADPH) oxidase as indicated by a marked stimulation of p67(phox)/rac1 NADPH oxidase subunit expression and by increased rac1 membrane association. In addition, superoxide production in rat heart mitochondria was up-regulated by sirolimus, as measured by L012-enhanced chemiluminescence. As a consequence, electron spin resonance measurements showed a 40% reduction in vascular nitric oxide bioavailability, which was further supported by decreased serum nitrite levels. CONCLUSIONS Sirolimus causes marked vascular dysfunction and nitrate resistance after continuous treatment for 7 days. This impaired vasorelaxation may, in part, be induced by up-regulated mitochondrial superoxide release as well as by an up-regulation of NADPH oxidase-driven superoxide production. Both processes could contribute to endothelial dysfunction observed after coronary vascular interventions with sirolimus-coated stents.

Heparin-polynitroxides: Synthesis and preliminary evaluation as cardiovascular EPR/MR imaging probes and extracellular space-targeted antioxidants

We report here the synthesis of heparin-polynitroxide derivatives (HPNs) in which nitroxide moieties are linked either to uronic acid or glycosamine residues of the heparin macromolecule. HPNs have low anticoagulant activity, possess superoxide scavenging properties, bind to the vascular endothelium/extra-cellular matrix and can be detected by EPR and MRI techniques. As the vascular wall-targeted redox-active paramagnetic compounds, HPNs may have both diagnostic (molecular MRI) and therapeutic (ecSOD mimics) applications.

Spin-Labeled Heparins as Polarizing Agents for Dynamic Nuclear Polarization

A potentially biocompatible class of spin-labeled macromolecules, spin-labeled (SL) heparins, and their use as nuclear magnetic resonance (NMR) signal enhancers are introduced. The signal enhancement is achieved through Overhauser-type dynamic nuclear polarization (DNP). All presented SL-heparins show high (1)H DNP enhancement factors up to E=-110, which validates that effectively more than one hyperfine line can be saturated even for spin-labeled polarizing agents. The parameters for the Overhauser-type DNP are determined and discussed. A striking result is that for spin-labeled heparins, the off-resonant electron paramagnetic resonance (EPR) hyperfine lines contribute a non-negligible part to the total saturation, even in the absence of Heisenberg spin exchange (HSE) and electron spin-nuclear spin relaxation (T(1ne)). As a result, we conclude that one can optimize the use of, for example, biomacromolecules for DNP, for which only small sample amounts are available, by using heterogeneously distributed radicals attached to the molecule.

Heparin-polynitroxide derivatives: a platform for new diagnostic and therapeutic agents in cardiovascular disease?

Vascular wall extracellular oxidative stress Cardiovascular disease (CVD; mainly atherosclerosis, hypertension and diabetes mellitus) remains a major cause of death in western society [1]. Despite substantial progress achieved, the diagnosis of CVD often comes too late, when the disease has already advanced to therapeutically incurable stages. The development of efficient diagnostic probes allowing early non-invasive diagnostics, as well as drugs which can prevent or reverse CVD and/or its complications (e.g., myocardium infarctus and stroke) are highly desired tasks of the modern cardiovascular medicinal chemistry.

Development of a Polarizer and Biocompatible Polarizing Agents for Use in Dynamic Nuclear Polarization DNP-Enhanced NMR and MRI

The application of 13C (or other low γ nuclei) NMR spectroscopy and imaging for clinical diagnosis has been constrained by the extremely long imaging and spectroscopy acquisition times that are required to obtain high SNR under physiological conditions (low natural abundance of 13C, low concentration of 13C-compounds, physiological temperature etc.). However, this obstacle could be overcome by in vitro hyperpolarization of a 13C-containing molecule with long spin lattice relaxation time via dynamic nuclear polarization (DNP) and subsequent injection into the animal or patient of investigation [1, 2]. DNP is achieved by resonant excitation of electron spins of radicals (electron paramagnetic resonance, EPR) that have to be employed.