Skip to Content
Merck
CN
  • PF-1355, a mechanism-based myeloperoxidase inhibitor, prevents immune complex vasculitis and anti-glomerular basement membrane glomerulonephritis.

PF-1355, a mechanism-based myeloperoxidase inhibitor, prevents immune complex vasculitis and anti-glomerular basement membrane glomerulonephritis.

The Journal of pharmacology and experimental therapeutics (2015-02-24)
Wei Zheng, Roscoe Warner, Roger Ruggeri, Chunyan Su, Christian Cortes, Athanasia Skoura, Jessica Ward, Kay Ahn, Amit Kalgutkar, Dexue Sun, Tristan S Maurer, Paul D Bonin, Carlin Okerberg, Walter Bobrowski, Thomas Kawabe, Yanwei Zhang, Timothy Coskran, Sammy Bell, Bhupesh Kapoor, Kent Johnson, Leonard Buckbinder
ABSTRACT

Small vessel vasculitis is a life-threatening condition and patients typically present with renal and pulmonary injury. Disease pathogenesis is associated with neutrophil accumulation, activation, and oxidative damage, the latter being driven in large part by myeloperoxidase (MPO), which generates hypochlorous acid among other oxidants. MPO has been associated with vasculitis, disseminated vascular inflammation typically involving pulmonary and renal microvasculature and often resulting in critical consequences. MPO contributes to vascular injury by 1) catabolizing nitric oxide, impairing vasomotor function; 2) causing oxidative damage to lipoproteins and endothelial cells, leading to atherosclerosis; and 3) stimulating formation of neutrophil extracellular traps, resulting in vessel occlusion and thrombosis. Here we report a selective 2-thiouracil mechanism-based MPO inhibitor (PF-1355 [2-(6-(2,5-dimethoxyphenyl)-4-oxo-2-thioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide) and demonstrate that MPO is a critical mediator of vasculitis in mouse disease models. A pharmacokinetic/pharmacodynamic response model of PF-1355 exposure in relation with MPO activity was derived from mouse peritonitis. The contribution of MPO activity to vasculitis was then examined in an immune complex model of pulmonary disease. Oral administration of PF-1355 reduced plasma MPO activity, vascular edema, neutrophil recruitment, and elevated circulating cytokines. In a model of anti-glomerular basement membrane disease, formerly known as Goodpasture disease, albuminuria and chronic renal dysfunction were completely suppressed by PF-1355 treatment. This study shows that MPO activity is critical in driving immune complex vasculitis and provides confidence in testing the hypothesis that MPO inhibition will provide benefit in treating human vasculitic diseases.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Hemin, from bovine, ≥90%
Sigma-Aldrich
Hydrogen peroxide solution, 34.5-36.5%
Sigma-Aldrich
Hemin, BioXtra, from Porcine, ≥96.0% (HPLC)
Sigma-Aldrich
Bicinchoninic acid disodium salt hydrate, ≥98% (HPLC)
USP
Doxycycline hyclate, United States Pharmacopeia (USP) Reference Standard
Supelco
Doxycycline hyclate, VETRANAL®, analytical standard
Sigma-Aldrich
Glutaraldehyde solution, 50 wt. % in H2O
Sigma-Aldrich
Glutaraldehyde solution, Grade II, 25% in H2O
Sigma-Aldrich
Glutaraldehyde solution, technical, ~50% in H2O (5.6 M)
Sigma-Aldrich
Glutaraldehyde solution, 50% in H2O, suitable for photographic applications
Sigma-Aldrich
Doxycycline hyclate
Doxycycline hyclate, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Tris(hydroxymethyl)aminomethane, ACS reagent, ≥99.8%
Sigma-Aldrich
Hydrogen peroxide solution, contains ~200 ppm acetanilide as stabilizer, 3 wt. % in H2O
Millipore
Hydrogen peroxide solution, 3%, suitable for microbiology
Sigma-Aldrich
Trizma® base, BioPerformance Certified, meets EP, USP testing specifications, suitable for cell culture, ≥99.9% (titration)
Sigma-Aldrich
Trizma® base, Primary Standard and Buffer, ≥99.9% (titration), crystalline
Sigma-Aldrich
Trizma® base, ≥99.0% (T)
Sigma-Aldrich
Tromethamine, meets USP testing specifications
Sigma-Aldrich
Trizma® base, BioXtra, pH 10.5-12.0 (1 M in H2O), ≥99.9% (titration)
Sigma-Aldrich
Trizma® base, 99.7% (T), puriss. p.a.
Sigma-Aldrich
Trizma® base, anhydrous, free-flowing, Redi-Dri, ≥99.9%
Sigma-Aldrich
Trizma® base, BioUltra, Molecular Biology, ≥99.8% (T)
Sigma-Aldrich
Trizma® base, ≥99.9% (titration), crystalline
Supelco
Trizma® base, reference material for titrimetry, certified by BAM, >99.5%
Sigma-Aldrich
Glutaric dialdehyde solution, 50 wt. % in H2O, FCC
Sigma-Aldrich
Bicinchoninic acid disodium salt hydrate, Vetec, reagent grade, 98%
Sigma-Aldrich
Glutaraldehyde solution, Grade I, 8% in H2O, specially purified for use as an electron microscopy fixative or other sophisticated use
Sigma-Aldrich
Trizma® base, Vetec, reagent grade, ≥99%
Sigma-Aldrich
Glutaraldehyde solution, Grade I, 70% in H2O, specially purified for use as an electron microscopy fixative or other sophisticated use