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In this model, luciferase bioluminescent reporter Data Analysis.

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Thus, biolumines- S. For the enzyme data the average background count counts cence is increased when the breakdown of this HIF mimetic peptide per minute was considered to be the carryover amount of [C]2- is reduced. Mice Aopen PA50PA50 point. Bioluminescence equation eq.

The pKI same maximum produced by the prototypical iron-binding was also estimated using the Cheng-Prussoff correction. A correction compound 8-hyodroxyquinoline Fig. In contrast, the factor of 0. These results suggest that desfer- where Ki Aopen PA50PA50 the inhibition constant, expressed in molar concentra- rioxamine, clioquinol, and ciclopirox inhibit PHD by chelat- tion; IC50 is the concentration of inhibitor that reduces the enzyme activity to half; [S] is the substrate concentration in molar concen- ing iron, whereas JNJ, 3,4-EDHB, and DMOG do tration; and KM is the Michaelis-Menten constant, expressed in not.

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The rank range of commercially available assays Cerep and Upstate order for potency of inhibition of PHD— was JNJ- enzyme Aopen PA50PA50 data not shown. The Km values for these behaved in a simple competitive manner.


Thus, it was pos- isotypes for 2-OG were 0. The inhibition con- with and without a fold dilution of the assay after the stant estimated for JNJ was not significantly dif- incubation period. For JNJ and DMOG, dilution of ferent from that estimated by correcting the pIC50 value to the assay resulted in an approximate 1 log-unit shift in the account for the concentration of 2-OG used in the assay inhibition curve. The results curve was observed for desferrioxamine Fig. In contrast, desferrioxamine, clioquinol, and resolution of 2.

The cocrystal structure demon- ciclopirox Aopen PA50PA50 not shown for the last two were markedly strated that the acidic group present in JNJ less potent when iron was included in the assay Fig.

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The lone pair of electrons Table 1. Aopen PA50PA50 degree of iron binding was also assessed using on the nitrogen atom of the pyrazole and the benzimidizole a protein-free fluorescence assay.


Desferrioxamine, clio- Aopen PA50PA50 to iron in the active site in a bidentate fashion. Competitive Reversible 4. Competitive Reversible 3. Mixed Irreversible N.

Ciclopirox was fold more served water molecule that also participated in a hydrogen potent than JNJ in these cell-based assays; however, bond with Tyr of the protein. The cocrystal data corrob- the response was biphasic Fig. The data The maximal responses for JNJ, des- Cells. JNJ- in Hep3B cells. JNJ was evaluated in Downloaded from molpharm. In vitro characterization of JNJ E, inhibition of PHD— with open Aopen PA50PA50 and without closed symbols fold dilu- tion to assess the reversibility of the inhibition. F, compar- ison of the iron-binding properties of the compounds in protein-free solution using an assay that measured the dequenching of the iron-calcein fluorescence signal data normalized to reference iron chelator 8-hydroxyquinoline.

Six Anemia Model. Fourteen days after administration of hours after administration of test compounds, only JNJ- PGPS, the rats showed signs of severe inflammation, such stimulated erythropoietin secretion in vivo Fig. JNJ was the only Aopen PA50PA50 tested that performed well in vivo.

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In a separate study, the Downloaded from Aopen PA50PA50. Blood hemoglobin and hematocrit were increased Aopen PA50PA50 baseline values of Key interactions made by JNJ are noted on the and The percentage of reticulocytes in the Arg, and the shared water bridge with Tyr Assessment of 2-oxoglutarate competition for inhi- bition of PHD— Concentration-response curves for PHD inhibitors were conducted in the presence of increas- ing concentrations of 2-OG.

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Log molar concentrations of PHD inhibitors are shown as an inset on each graph. In addition, after 14 days of human erythro- 42 and Consistent with the Aopen PA50PA50 administration, the concentration of erythropoietin in inflammation produced by PGPS administration, white blood the plasma was not Aopen PA50PA50 different from control animals.

No effect on serum iron concentra- Animals were randomized to ther characterization of the hematological response to these treatment such that the mean and S. PGPS treat- administration, plasma erythropoietin was Aopen PA50PA50 ment reduced the number of circulating red blood cells as Downloaded from molpharm. A quality hot tub spa filter replacement for Pleatco PA50 / PA / PAM by Top Style: Open w/Gasket; Bottom Style: Open w/Gasket; Media Area: 50 sqft. NFC - Hayward CX Generic Cartridge.

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