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1.
Bioorg Med Chem Lett ; 24(3): 917-22, 2014 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-24412110

RESUMEN

The optimization of a novel series of non-nucleoside reverse transcriptase inhibitors (NNRTI) led to the identification of pyridone 36. In cell cultures, this new NNRTI shows a superior potency profile against a range of wild type and clinically relevant, resistant mutant HIV viruses. The overall favorable preclinical pharmacokinetic profile of 36 led to the prediction of a once daily low dose regimen in human. NNRTI 36, now known as MK-1439, is currently in clinical development for the treatment of HIV infection.


Asunto(s)
Fármacos Anti-VIH/farmacología , Descubrimiento de Drogas , Farmacorresistencia Viral/efectos de los fármacos , VIH-1/efectos de los fármacos , Piridonas/química , Piridonas/farmacología , Inhibidores de la Transcriptasa Inversa/síntesis química , Inhibidores de la Transcriptasa Inversa/farmacología , Triazoles/química , Triazoles/farmacología , Animales , Fármacos Anti-VIH/síntesis química , Fármacos Anti-VIH/química , Células Cultivadas , Cristalografía por Rayos X , Perros , VIH-1/genética , Humanos , Concentración 50 Inhibidora , Estructura Molecular , Mutación , Ratas , Ratas Sprague-Dawley , Inhibidores de la Transcriptasa Inversa/química
2.
Am J Respir Cell Mol Biol ; 45(1): 81-7, 2011 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-20855652

RESUMEN

Cathepsin S (Cat S) is predominantly expressed in antigen-presenting cells and is up-regulated in several preclinical models of antigen-induced inflammation, suggesting a role in the allergic response. Prophylactic dosing of an irreversible Cat S inhibitor has been shown to attenuate pulmonary eosinophilia in mice, supporting the hypothesis that Cat S inhibition before the initiation of airway inflammation is beneficial in airway disease. In addition, Cat S has been shown to play a role in more distal events in the allergic response. To determine where Cat S inhibition may affect the allergic response, we used complementary genetic and pharmacological approaches to investigate the role of Cat S in the early and downstream allergic events in a murine model of antigen-induced lung inflammation. Cat S knockout mice did not develop ovalbumin-induced pulmonary inflammation, consistent with a role for Cat S in the development of the allergic response. Alternatively, wild-type mice were treated with a reversible, highly selective Cat S inhibitor in prophylactic and therapeutic dosing paradigms and assessed for changes in airway inflammation. Although both treatment paradigms resulted in potent Cat S inhibition, only prophylactic Cat S inhibitor dosing blocked lung inflammation, consistent with our findings in Cat S knockout mice. The findings indicate that although Cat S is up-regulated in allergic models, it does not appear to play a significant role in the downstream effector inflammatory phase in this model; however, our results demonstrate that Cat S inhibition in a prophylactic paradigm would ameliorate airway inflammation.


Asunto(s)
Asma/prevención & control , Catepsinas/genética , Catepsinas/farmacología , Animales , Asma/genética , Asma/metabolismo , Catepsinas/biosíntesis , Modelos Animales de Enfermedad , Evaluación de Medicamentos , Humanos , Ratones , Ratones Noqueados , Ovalbúmina/efectos adversos , Ovalbúmina/farmacología , Eosinofilia Pulmonar/genética , Eosinofilia Pulmonar/metabolismo , Eosinofilia Pulmonar/prevención & control , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/genética
3.
Biol Chem ; 391(12): 1469-73, 2010 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-20868234

RESUMEN

Renin is the first enzyme in the renin-angiotensin-aldosterone system which is the principal regulator of blood pressure and hydroelectrolyte balance. Previous studies suggest that cathepsin B is the activator of the prorenin zymogen. Here, we show no difference in plasma renin activity, or mean arterial blood pressure between wild-type and cathepsin B knockout mice. To account for potential gene compensation, a potent, selective, reversible cathepsin B inhibitor was developed to determine the role of cathepsin B on prorenin processing in rats. Pharmacological inhibition of cathepsin B in spontaneously hypertensive and double transgenic rats did not result in a reduction in renal mature renin protein levels or plasma renin activity. We conclude that cathepsin B does not play a significant role in this process in rodents.


Asunto(s)
Catepsina B/fisiología , Renina/metabolismo , Animales , Catepsina B/antagonistas & inhibidores , Catepsina B/genética , Inhibidores Enzimáticos/farmacología , Hipertensión/genética , Hipertensión/metabolismo , Ratones , Ratones Noqueados , Ratas , Ratas Transgénicas
4.
Bioorg Med Chem Lett ; 20(3): 887-92, 2010 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-20061146

RESUMEN

MK-0674 is a potent and selective cathepsin K inhibitor from the same structural class as odanacatib with a comparable inhibitory potency profile against Cat K. It is orally bioavailable and exhibits long half-life in pre-clinical species. In vivo studies using deuterated MK-0674 show stereoselective epimerization of the alcohol stereocenter via an oxidation/reduction cycle. From in vitro incubations, two metabolites could be identified: the hydroxyleucine and the glucuronide conjugate which were confirmed using authentic synthetic standards.


Asunto(s)
Compuestos de Bifenilo/administración & dosificación , Compuestos de Bifenilo/farmacocinética , Catepsina K/antagonistas & inhibidores , Inhibidores de Cisteína Proteinasa/administración & dosificación , Inhibidores de Cisteína Proteinasa/farmacocinética , Descubrimiento de Drogas/métodos , Administración Oral , Animales , Disponibilidad Biológica , Compuestos de Bifenilo/química , Catepsina K/metabolismo , Inhibidores de Cisteína Proteinasa/química , Perros , Hepatocitos/efectos de los fármacos , Hepatocitos/metabolismo , Humanos , Macaca mulatta , Conejos , Ratas
5.
Bioorg Med Chem Lett ; 18(3): 923-8, 2008 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-18226527

RESUMEN

Odanacatib is a potent, selective, and neutral cathepsin K inhibitor which was developed to address the metabolic liabilities of the Cat K inhibitor L-873724. Substituting P1 and modifying the P2 side chain led to a metabolically robust inhibitor with a long half-life in preclinical species. Odanacatib was more selective in whole cell assays than the published Cat K inhibitors balicatib and relacatib. Evaluation in dermal fibroblast culture showed minimal intracellular collagen accumulation relative to less selective Cat K inhibitors.


Asunto(s)
Compuestos de Bifenilo/farmacología , Catepsinas/antagonistas & inhibidores , Inhibidores de Cisteína Proteinasa/farmacología , Animales , Azepinas/química , Azepinas/farmacología , Catepsina K , Colágeno/efectos de los fármacos , Colágeno/inmunología , Perros , Fibroblastos/efectos de los fármacos , Humanos , Modelos Biológicos , Estructura Molecular , Osteoporosis Posmenopáusica/tratamiento farmacológico , Piel/citología , Relación Estructura-Actividad , Sulfonas/química , Sulfonas/farmacología
6.
Biochem Pharmacol ; 72(10): 1279-92, 2006 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-16956584

RESUMEN

Insulin binds to the alpha subunit of the insulin receptor (IR) on the cell surface. The insulin-IR complex is subsequently internalized and trafficked within the cell. Endocytosed receptors, devoid of insulin, recycle back to the plasma membrane through the endocytic recycling compartment (ERC). Using a high content screening system, we investigate the intracellular trafficking of the IR and its phosphorylation state, within the ERC, in response to protein tyrosine phosphatase-1B (PTP1B) inhibition. Insulin stimulates, in a time- and dose-dependent manner, the accumulation of phosphorylated IR (pY(1158,1162,1163 IR) in the ERC of CHO-IR cells. Treatment of CHO-IR cells with PTP1B-specific inhibitors or siRNA leads to dose-dependent increases in IR residency and phosphorylation within the ERC. The results also demonstrate that PTP1B redistributes within CHO-IR cells upon insulin challenge. The established system will allow for efficient screening of candidate inhibitors for the modulation of PTP1B activity.


Asunto(s)
Endosomas/metabolismo , Insulina/farmacología , Proteínas Tirosina Fosfatasas/fisiología , Receptor de Insulina/metabolismo , Animales , Línea Celular , Cricetinae , Cricetulus , Relación Dosis-Respuesta a Droga , Endocitosis/efectos de los fármacos , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Humanos , Estructura Molecular , Fosforilación , Transporte de Proteínas , Proteína Tirosina Fosfatasa no Receptora Tipo 1 , Proteínas Tirosina Fosfatasas/antagonistas & inhibidores , Factores de Tiempo
7.
J Med Chem ; 48(24): 7535-43, 2005 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-16302795

RESUMEN

The lysosomal cysteine protease cathepsin K is a target for osteoporosis therapy. The aryl-piperazine-containing cathepsin K inhibitor CRA-013783/L-006235 (1) displays greater than 4000-fold selectivity against the lysosomal/endosomal antitargets cathepsin B, L, and S. However, 1 and other aryl-piperazine-containing analogues, including balicatib (10), are approximately 10-100-fold more potent in cell-based enzyme occupancy assays than against each purified enzyme. This phenomenon arises from their basic, lipophilic nature, which results in lysosomal trapping. Consistent with its lysosomotropic nature, 1 accumulates in cells and in rat tissues of high lysosome content. In contrast, nonbasic aryl-morpholino-containing analogues do not exhibit lysosomotropic properties. Increased off-target activities of basic cathepsin K inhibitors were observed in a cell-based cathepsin S antigen presentation assay. No potency increases of basic inhibitors in a functional cathepsin K bone resorption whole cell assay were detected. Therefore, basic cathepsin K inhibitors, such as 1, suffer from reduced functional selectivities compared to those predicted using purified enzyme assays.


Asunto(s)
Benzamidas/farmacología , Catepsinas/antagonistas & inhibidores , Lisosomas/efectos de los fármacos , Morfolinas/farmacología , Piperazinas/farmacología , Tiazoles/farmacología , Animales , Presentación de Antígeno/efectos de los fármacos , Autorradiografía , Benzamidas/química , Benzamidas/farmacocinética , Conservadores de la Densidad Ósea/química , Conservadores de la Densidad Ósea/farmacocinética , Conservadores de la Densidad Ósea/farmacología , Catepsina B/antagonistas & inhibidores , Catepsina K , Catepsina L , Línea Celular , Cisteína Endopeptidasas , Femenino , Humanos , Lisosomas/enzimología , Ratones , Ratones Endogámicos C57BL , Morfolinas/química , Piperazinas/química , Piperazinas/farmacocinética , Conejos , Ratas , Relación Estructura-Actividad , Tiazoles/química , Tiazoles/farmacocinética , Distribución Tisular
8.
J Renin Angiotensin Aldosterone Syst ; 12(3): 133-45, 2011 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-21393355

RESUMEN

INTRODUCTION: The hypertensive double-transgenic (dTG) rat strain, expressing human renin and angiotensinogen, develops severe hypertension and organ damage and 50% of individuals die by 7 weeks of age. Here, we characterise a variation of this model in which animals present stable hypertension. MATERIALS AND METHODS: The effect of renin-angiotensin system blockers on blood pressure was determined with adult dTG rats treated with enalapril from 3 to 12 weeks of age. Tissue expression levels of renin and angiotensinogen were determined in dTG rats and rhesus monkeys by quantitative PCR. RESULTS: Upon withdrawal from enalapril, mean arterial pressure (MAP) rose to 160-180 mmHg, with 95% of the female dTG rats surviving for 6 to 12 months, In Sprague-Dawley (SD) rats and rhesus monkeys, renin mRNA was absent or weakly expressed in most tissues, except for the kidneys and adrenals. In dTG rats, human renin expression was high in many additional tissues. The expression of human angiotensinogen in dTG rats followed a similar tissue pattern to SD and rhesus monkey angiotensinogen. Oral dosing of aliskiren, enalapril or losartan provided a similar maximal reduction in MAP and duration of efficacy in telemetrised dTG rats. CONCLUSIONS: Enalapril-pretreated dTG rats are suitable for long-term MAP monitoring and sequential evaluation of human renin inhibitors.


Asunto(s)
Enalapril/farmacología , Enalapril/uso terapéutico , Hipertensión/tratamiento farmacológico , Renina/antagonistas & inhibidores , Amidas/administración & dosificación , Amidas/farmacología , Amidas/uso terapéutico , Angiotensinógeno/genética , Angiotensinógeno/metabolismo , Animales , Presión Sanguínea/efectos de los fármacos , Modelos Animales de Enfermedad , Enalapril/administración & dosificación , Femenino , Fumaratos/administración & dosificación , Fumaratos/farmacología , Fumaratos/uso terapéutico , Regulación de la Expresión Génica/efectos de los fármacos , Frecuencia Cardíaca/efectos de los fármacos , Humanos , Hipertensión/fisiopatología , Macaca mulatta , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-Dawley , Ratas Transgénicas , Renina/sangre , Renina/genética , Distribución Tisular/efectos de los fármacos
9.
J Biol Chem ; 282(42): 30423-33, 2007 Oct 19.
Artículo en Inglés | MEDLINE | ID: mdl-17664276

RESUMEN

Although protein-tyrosine phosphatase 1B (PTP-1B) is a negative regulator of insulin action, adipose tissue from PTP-1B-/- mice does not show enhanced insulin-stimulated insulin receptor phosphorylation. Investigation of glucose uptake in isolated adipocytes revealed that the adipocytes from PTP-1B-/- mice have a significantly attenuated insulin response as compared with PTP-1B+/+ adipocytes. This insulin resistance manifests in PTP-1B-/- animals older than 16 weeks of age and could be partially rescued by adenoviral expression of PTP-1B in null adipocytes. Examination of adipose signaling pathways found that the basal p70S6K activity was at least 50% higher in adipose from PTP-1B-/- mice compared with wild type animals. The increased basal activity of p70S6K in PTP-1B-/- adipose correlated with decreases in IR substrate-1 protein levels and insulin-stimulated Akt/protein kinase B activity, explaining the decrease in insulin sensitivity even as insulin receptor phosphorylation was unaffected. The insulin resistance of the of the PTP-1B-/- adipocytes could also be rescued by treatment with rapamycin, suggesting that in adipose the loss of PTP-1B results in basal activation of mTOR (mammalian target of rapamycin) complex 1 leading to a tissue-specific insulin resistance.


Asunto(s)
Adipocitos/enzimología , Tejido Adiposo/enzimología , Resistencia a la Insulina/genética , Proteína Tirosina Fosfatasa no Receptora Tipo 1/deficiencia , Proteínas Quinasas S6 Ribosómicas 70-kDa/metabolismo , Adenoviridae , Adipocitos/patología , Tejido Adiposo/patología , Animales , Antibióticos Antineoplásicos/farmacología , Activación Enzimática/efectos de los fármacos , Activación Enzimática/genética , Glucosa/metabolismo , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados , Especificidad de Órganos/genética , Fosforilación , Proteínas Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Receptor de Insulina/metabolismo , Sirolimus/farmacología , Serina-Treonina Quinasas TOR
11.
Bioorg Med Chem Lett ; 13(6): 1195-8, 2003 Mar 24.
Artículo en Inglés | MEDLINE | ID: mdl-12643942
12.
Anal Biochem ; 335(2): 218-27, 2004 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-15556560

RESUMEN

We describe a novel diazomethylketone-containing irreversible inhibitor (BIL-DMK) which is specific for a subset of pharmaceutically important cysteine cathepsin proteases. BIL-DMK rapidly inactivates cathepsins B, F, K, L, S, and V in isolated enzyme assays and labels cathepsins in whole cells. The presence of catalytically active cathepsins B, L, and K or S was demonstrated using radioiodinated BIL-DMK in HepG2 (hepatoma), HIG82 (rabbit synoviocyte), and Ramos (B lymphoma) cell lines, respectively. The identity of each protein labeled was confirmed from the isoelectric point and molecular mass of the radioactive spots on two-dimensional gel and by comigration with each cathepsin as identified by immunoblotting. These cell lines were used to establish whole-cell enzyme occupancy assays to determine the potency of both irreversible and reversible inhibitors against each cathepsin in their native cellular lysosomal or endosomal environment. These whole-cell enzyme occupancy assays are useful to determine the cellular permeability of competing inhibitors and have the advantage of not requiring specific substrates for each cathepsin of interest.


Asunto(s)
Compuestos de Bifenilo/farmacología , Catepsinas/metabolismo , Inhibidores de Cisteína Proteinasa/farmacología , Diazometano/análogos & derivados , Leucina/análogos & derivados , Animales , Autorradiografía , Western Blotting , Catepsina B/análisis , Catepsina B/antagonistas & inhibidores , Catepsina K , Catepsina L , Catepsinas/análisis , Catepsinas/antagonistas & inhibidores , Línea Celular Tumoral , Cisteína Endopeptidasas/análisis , Diazometano/síntesis química , Diazometano/farmacología , Humanos , Radioisótopos de Yodo , Leucina/farmacología , Conejos
13.
Bioorg Med Chem Lett ; 14(4): 1039-42, 2004 Feb 23.
Artículo en Inglés | MEDLINE | ID: mdl-15013019

RESUMEN

The SAR from our peptide libraries was exploited to design a series of potent deoxybenzoin PTP-1B inhibitors. The introduction of an ortho bromo substituent next to the difluoromethylphosphonate warhead gave up to 20-fold increase in potency compared to the desbromo analogues. In addition, these compounds were orally bioavailable and active in the animal models of non-insulin dependent diabetes mellitus (NIDDM).


Asunto(s)
Benzoína/farmacología , Inhibidores Enzimáticos/farmacología , Proteínas Tirosina Fosfatasas/antagonistas & inhibidores , Administración Oral , Animales , Benzoína/análogos & derivados , Benzoína/síntesis química , Disponibilidad Biológica , Línea Celular , Supervivencia Celular/efectos de los fármacos , Diabetes Mellitus/enzimología , Diabetes Mellitus/metabolismo , Diseño de Fármacos , Inhibidores Enzimáticos/síntesis química , Insectos , Ratones , Ratones Noqueados , Modelos Animales , Proteína Tirosina Fosfatasa no Receptora Tipo 1 , Relación Estructura-Actividad
15.
Biochemistry ; 42(39): 11451-9, 2003 Oct 07.
Artículo en Inglés | MEDLINE | ID: mdl-14516196

RESUMEN

Protein tyrosine phosphatase 1B (PTP1B) has been implicated in the regulation of the insulin signaling pathway and represents an attractive target for the design of inhibitors in the treatment of type 2 diabetes and obesity. Inspection of the structure of PTP1B indicates that potent PTP1B inhibitors may be obtained by targeting a secondary aryl phosphate-binding site as well as the catalytic site. We report here the crystal structures of PTP1B in complex with first and second generation aryldifluoromethyl-phosphonic acid inhibitors. While all compounds bind in a previously unexploited binding pocket near the primary binding site, the second generation compounds also reach into the secondary binding site, and exhibit moderate selectivity for PTP1B over the closely related T-cell phosphatase. The molecular basis for the selectivity has been confirmed by single point mutation at position 52, where the two phosphatases differ by a phenylalanine-to-tyrosine switch. These compounds present a novel platform for the development of potent and selective PTP1B inhibitors.


Asunto(s)
Proteínas Tirosina Fosfatasas/antagonistas & inhibidores , Proteínas Tirosina Fosfatasas/química , Triazoles/química , Triazoles/farmacología , Aminoácidos/genética , Aminoácidos/metabolismo , Sitios de Unión , Dominio Catalítico , Cristalografía por Rayos X , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Humanos , Enlace de Hidrógeno , Concentración 50 Inhibidora , Modelos Moleculares , Ácidos Fosfínicos/química , Ácidos Fosfínicos/metabolismo , Ácidos Fosfínicos/farmacología , Mutación Puntual , Proteína Tirosina Fosfatasa no Receptora Tipo 1 , Proteínas Tirosina Fosfatasas/genética , Proteínas Tirosina Fosfatasas/metabolismo , Proteínas Recombinantes/antagonistas & inhibidores , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Relación Estructura-Actividad , Especificidad por Sustrato
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