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1.
Bioorg Med Chem Lett ; 23(9): 2590-4, 2013 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-23535330

RESUMEN

Drug design efforts in the emerging 2-aminothiazole-4-carboxamide class of CHK1 inhibitors have uncovered specific combinations of key substructures within the molecule; resulting in significant improvements in cell-based activity while retaining a greater than one hundred-fold selectivity against CDK2. The X-ray crystal structure of a complex between compound 39 and the CHK1 protein detailing a 'U-shaped' topology and key interactions with the protein surface at the ATP site is also reported.


Asunto(s)
Amidas/química , Diseño de Fármacos , Inhibidores de Proteínas Quinasas/química , Proteínas Quinasas/química , Tiazoles/química , Amidas/síntesis química , Amidas/metabolismo , Sitios de Unión , Quinasa 1 Reguladora del Ciclo Celular (Checkpoint 1) , Cristalografía por Rayos X , Quinasa 2 Dependiente de la Ciclina/antagonistas & inhibidores , Quinasa 2 Dependiente de la Ciclina/metabolismo , Simulación del Acoplamiento Molecular , Unión Proteica , Inhibidores de Proteínas Quinasas/síntesis química , Inhibidores de Proteínas Quinasas/metabolismo , Proteínas Quinasas/metabolismo , Relación Estructura-Actividad
2.
J Lipid Res ; 52(4): 646-56, 2011 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-21296956

RESUMEN

Fatty acid binding protein-4 (FABP4) and FABP5 are two closely related FA binding proteins expressed primarily in adipose tissue and/or macrophages. The small-molecule FABP4 inhibitor BMS309403 was previously reported to improve insulin sensitivity in leptin-deficient Lep(ob)/Lep(ob) (ob/ob) mice. However, this compound was not extensively characterized in the more physiologically relevant animal model of mice with diet-induced obesity (DIO). Here, we report the discovery and characterization of a novel series of FABP4/5 dual inhibitors represented by Compounds 1-3. Compared with BMS309403, the compounds had significant in vitro potency toward both FABP4 and FABP5. In cell-based assays, Compounds 2 and 3 were more potent than BMS309403 to inhibit lipolysis in 3T3-L1 adipocytes and in primary human adipocytes. They also inhibited MCP-1 release from THP-1 macrophages as well as from primary human macrophages. When chronically administered to DIO mice, BMS309403 and Compound 3 reduced plasma triglyceride and free FA levels. Compound 3 reduced plasma free FAs at a lower dose level than BMS309403. However, no significant change was observed in insulin, glucose, or glucose tolerance. Our results indicate that the FABP4/5 inhibitors ameliorate dyslipidemia but not insulin resistance in DIO mice.


Asunto(s)
Grasas de la Dieta/efectos adversos , Proteínas de Unión a Ácidos Grasos/antagonistas & inhibidores , Hipolipemiantes/uso terapéutico , Proteínas de Neoplasias/antagonistas & inhibidores , Obesidad/tratamiento farmacológico , Células 3T3-L1 , Adipocitos/efectos de los fármacos , Adipocitos/metabolismo , Animales , Células Cultivadas , Quimiocina CCL2/metabolismo , Dislipidemias/inducido químicamente , Dislipidemias/tratamiento farmacológico , Ácidos Grasos no Esterificados/sangre , Resistencia a la Insulina , Lipólisis/efectos de los fármacos , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Ratones , Obesidad/inducido químicamente , Triglicéridos/sangre
3.
Bioorg Med Chem Lett ; 20(7): 2119-24, 2010 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-20219368

RESUMEN

SAR exploration from an initial hit, (S)-N-(2-cyclohexenylethyl)-2-fluoro-6-(2-(1-hydroxy-3-phenylpropan-2-ylamino)-2-oxoethoxy)benzamide (1), identified using our proprietary automated ligand identification system (ALIS),(1) has led to a novel series of selective hepatitis C virus (HCV) NS5B polymerase inhibitors with improved in vitro potency as exemplified by (S)-2-fluoro-6-(2-(1-hydroxy-3-phenylpropan-2-ylamino)-2-oxoethoxy)-N-isopentyl-N-methylbenzamidecarboxamide (41) (IC(50)=0.5 microM). The crystal structure of an analogue (44) was solved and provided rationalization of the SAR of this series, which binds in a distinct manner in the palm domain of NS5B, consistent with biochemical analysis using enzyme mutant variants. These data warrant further lead optimization efforts on this novel series of non-nucleoside inhibitors targeting the HCV polymerase.


Asunto(s)
Benzamidas/química , Benzamidas/farmacología , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Hepacivirus/enzimología , Proteínas no Estructurales Virales/antagonistas & inhibidores , Proteínas no Estructurales Virales/metabolismo , Antivirales/química , Antivirales/farmacología , Cristalografía , Diseño de Fármacos , Hepatitis C/tratamiento farmacológico , Humanos , Modelos Moleculares , ARN Polimerasa Dependiente del ARN/antagonistas & inhibidores , ARN Polimerasa Dependiente del ARN/química , ARN Polimerasa Dependiente del ARN/metabolismo , Relación Estructura-Actividad , Proteínas no Estructurales Virales/química
4.
Bioorg Med Chem Lett ; 19(23): 6507-14, 2009 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-19875284

RESUMEN

The biotin carboxylase (AccC) is part of the multi-component bacterial acetyl coenzyme-A carboxylase (ACCase) and is essential for pathogen survival. We describe herein the affinity optimization of an initial hit to give 2-(2-chlorobenzylamino)-1-(cyclohexylmethyl)-1H-benzo[d]imidazole-5-carboxamide (1), which was identified using our proprietary Automated Ligand Identification System (ALIS).(1) The X-ray co-crystal structure of 1 was solved and revealed several key interactions and opportunities for further optimization in the ATP site of AccC. Structure Based Drug Design (SBDD) and parallel synthetic approaches resulted in a novel series of AccC inhibitors, exemplified by (R)-2-(2-chlorobenzylamino)-1-(2,3-dihydro-1H-inden-1-yl)-1H-imidazo[4,5-b]pyridine-5-carboxamide (40). This compound is a potent and selective inhibitor of bacterial AccC with an IC(50) of 20 nM and a MIC of 0.8 microg/mL against a sensitized strain of Escherichia coli (HS294 E. coli).


Asunto(s)
Antibacterianos/farmacología , Bencimidazoles/farmacología , Ligasas de Carbono-Nitrógeno/antagonistas & inhibidores , Descubrimiento de Drogas , Inhibidores Enzimáticos/farmacología , Escherichia coli/efectos de los fármacos , Imidazoles/farmacología , Ácidos Nicotínicos/farmacología , Antibacterianos/síntesis química , Antibacterianos/química , Bencimidazoles/síntesis química , Bencimidazoles/química , Cristalografía por Rayos X , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/química , Imidazoles/síntesis química , Imidazoles/química , Ligandos , Pruebas de Sensibilidad Microbiana , Modelos Moleculares , Ácidos Nicotínicos/síntesis química , Ácidos Nicotínicos/química , Relación Estructura-Actividad
5.
J Biomol Screen ; 11(2): 194-207, 2006 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-16490772

RESUMEN

Screening assays using target-based affinity selection coupled with high-sensitivity detection technologies to identify small-molecule hits from chemical libraries can provide a useful discovery approach that complements traditional assay systems. Affinity selection-mass spectrometry (AS-MS) is one such methodology that holds promise for providing selective and sensitive high-throughput screening platforms. Although AS-MS screening platforms have been used to discover small-molecule ligands of proteins from many target families, they have not yet been used routinely to screen integral membrane proteins. The authors present a proof-of-concept study using size exclusion chromatography coupled to AS-MS to perform a primary screen for small-molecule ligands of the purified muscarinic M2 acetylcholine receptor, a G-protein-coupled receptor. AS-MS is used to characterize the binding mechanisms of 2 newly discovered ligands. NGD-3350 is a novel M2-specific orthosteric antagonist of M2 function. NGD-3366 is an allosteric ligand with binding properties similar to the allosteric antagonist W-84, which decreases the dissociation rate of N-methyl-scopolamine from the M2 receptor. Binding properties of the ligands discerned from AS-MS assays agree with those from in vitro biochemical assays. The authors conclude that when used with appropriate small-molecule libraries, AS-MS may provide a useful high-throughput assay system for the discovery and characterization of all classes of integral membrane protein ligands, including allosteric modulators.


Asunto(s)
Espectrometría de Masas/métodos , Antagonistas Muscarínicos/farmacología , Unión Proteica/efectos de los fármacos , Receptor Muscarínico M2/química , Receptores Acoplados a Proteínas G/antagonistas & inhibidores , Acetilcolina , Regulación Alostérica , Sitio Alostérico , Animales , Línea Celular , Relación Dosis-Respuesta a Droga , Cobayas , Insectos , Ligandos , Estructura Molecular , Ensayo de Unión Radioligante , Receptor Muscarínico M2/aislamiento & purificación
6.
ACS Med Chem Lett ; 3(2): 123-8, 2012 Feb 09.
Artículo en Inglés | MEDLINE | ID: mdl-24900442

RESUMEN

A novel series of CHK1 inhibitors with a distinctive hinge binding mode, exemplified by 2-aryl-N-(2-(piperazin-1-yl)phenyl)thiazole-4-carboxamide, was discovered through high-throughput screening using the affinity selection-mass spectrometry (AS-MS)-based Automated Ligand Identification System (ALIS) platform. Structure-based ligand design and optimization led to significant improvements in potency to the single digit nanomolar range and hundred-fold selectivity against CDK2.

7.
Comb Chem High Throughput Screen ; 15(6): 473-85, 2012 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-22651846

RESUMEN

Affinity selection-mass spectrometry (AS-MS) is a sensitive technology for identifying small molecules that bind to target proteins, and assays enabled by AS-MS can be used to delineate relative binding affinities of ligands for proteins. 'Indirect' AS-MS assays employ size-exclusion techniques to separate target-ligand complexes from unbound ligands, and target-associated ligands are then specifically detected by liquid chromatography mass spectrometry. We report how indirect AS-MS binding assays with known reference control compounds were used as guideposts for development of an optimized purification method for CXCR4, a G-protein coupled chemokine receptor, for which we sought novel antagonists. The CXCR4 purification method that was developed was amenable to scale-up and enabled the screening of purified recombinant human CXCR4 against a large combinatorial library of small molecules by high throughput indirect AS-MS. The screen resulted in the discovery of new ligands that competed off binding of reference compounds to CXCR4 in AS-MS binding assays and that antagonized SDF1α-triggered responses and CXCR4-mediated HIV1 viral uptake in cell-based assays. This report provides a methodological paradigm whereby indirect AS-MS-based ligand binding assays may be used to guide optimal integral membrane protein purification methods that enable downstream affinity selection-based applications such as high throughput AS-MS screens.


Asunto(s)
Ensayos Analíticos de Alto Rendimiento/métodos , Espectrometría de Masas/métodos , Receptores CXCR4/aislamiento & purificación , Receptores CXCR4/metabolismo , Línea Celular Tumoral , Quimiocina CXCL12/metabolismo , Humanos , Ligandos , Unión Proteica , Receptores CXCR4/antagonistas & inhibidores
8.
ACS Med Chem Lett ; 2(8): 632-7, 2011 Aug 11.
Artículo en Inglés | MEDLINE | ID: mdl-24900358

RESUMEN

A novel series of non-ATP-competitive MK2 inhibitors based on a furan-2-carboxyamide scaffold was discovered through high-throughput screening using the affinity selection-mass spectrometry-based Automated Ligand Identification System platform. Medicinal chemistry efforts optimized the initial screening hit to leadlike compounds with significant improvements in biochemical and cellular potencies, while maintaining excellent kinase selectivity and in vitro pharmacokinetic properties. Biophysical and biochemical studies confirmed the unique non-ATP-competitive binding mode of this series and suggested that highly selective inhibitors of MK2 should be feasible by targeting the outside ATP pocket.

9.
ACS Med Chem Lett ; 1(9): 466-71, 2010 Dec 09.
Artículo en Inglés | MEDLINE | ID: mdl-24900232

RESUMEN

Pyridine carboxamide-based inhibitors of the hepatitis C virus (HCV) NS5B polymerase were diversified and optimized to a variety of topologically related scaffolds. In particular, the 2-methyl nicotinic acid scaffold was developed into inhibitors with improved biochemical (IC50-GT1b = 0.014 µM) and cell-based HCV replicon potency (EC50-GT1b = 0.7 µM). Biophysical and biochemical characterization identified this novel series of compounds as palm site binders to HCV polymerase.

10.
Comb Chem High Throughput Screen ; 12(8): 760-71, 2009 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-19531013

RESUMEN

This manuscript describes the discovery and characterization of inhibitors of the lipid phosphatase SHIP2, an important target for the treatment of Type 2 diabetes, using the Automated Ligand Identification System. ALIS is an affinity selection-mass spectrometry platform for label-free, high throughput screening of mixture-based combinatorial libraries. We detail the mass-encoded synthesis of a library that yielded NGD-61338, a pyrazole-based SHIP2 inhibitor. Quantitative ALIS affinity measurements and inhibition of SHIP2 enzymatic activity indicate that this compound has micromolar binding affinity and inhibitory activity for this target. This inhibitor, which does not contain a phosphatase "warhead," binds the active site of SHIP2 as determined by ALIS-based competition experiments with the enzyme's natural substrate, phosphatidylinositol 3,4,5-triphosphate (PIP3). Structure-activity relationships for NGD-61338 and two other ligand classes discovered by ALIS screening were explored using a combination of combinatorial library synthesis and ALIS-enabled affinity ranking in compound mixtures.


Asunto(s)
Técnicas Químicas Combinatorias , Inhibidores Enzimáticos/análisis , Inhibidores Enzimáticos/química , Ensayos Analíticos de Alto Rendimiento/métodos , Espectrometría de Masas/métodos , Monoéster Fosfórico Hidrolasas/antagonistas & inhibidores , Monoéster Fosfórico Hidrolasas/metabolismo , Dominio Catalítico , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/farmacología , Inositol Polifosfato 5-Fosfatasas , Estructura Molecular , Pirazoles/análisis , Pirazoles/síntesis química , Pirazoles/química , Pirazoles/farmacología , Estereoisomerismo , Relación Estructura-Actividad
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