RESUMEN
Fragment-based drug design has introduced a bottom-up process for drug development, with improved sampling of chemical space and increased effectiveness in early drug discovery. Here, we combine the use of pharmacophores, the most general concept of representing drug-target interactions with the theory of protein hotspots, to develop a design protocol for fragment libraries. The SpotXplorer approach compiles small fragment libraries that maximize the coverage of experimentally confirmed binding pharmacophores at the most preferred hotspots. The efficiency of this approach is demonstrated with a pilot library of 96 fragment-sized compounds (SpotXplorer0) that is validated on popular target classes and emerging drug targets. Biochemical screening against a set of GPCRs and proteases retrieves compounds containing an average of 70% of known pharmacophores for these targets. More importantly, SpotXplorer0 screening identifies confirmed hits against recently established challenging targets such as the histone methyltransferase SETD2, the main protease (3CLPro) and the NSP3 macrodomain of SARS-CoV-2.
Asunto(s)
Proteasas 3C de Coronavirus/química , Proteasas Similares a la Papaína de Coronavirus/química , Desarrollo de Medicamentos/métodos , Descubrimiento de Drogas/métodos , Ensayos Analíticos de Alto Rendimiento/métodos , N-Metiltransferasa de Histona-Lisina/química , Animales , Supervivencia Celular , Chlorocebus aethiops , Química Computacional , Cristalografía por Rayos X , Bases de Datos de Proteínas , Diseño de Fármacos , Células HEK293 , Humanos , Enlace de Hidrógeno , Interacciones Hidrofóbicas e Hidrofílicas , Ligandos , Unión Proteica , Receptores Acoplados a Proteínas G/química , SARS-CoV-2/química , SARS-CoV-2/genética , Bibliotecas de Moléculas Pequeñas , Células VeroRESUMEN
The discovery of novel uracil phenylethylamines bearing a butyric acid as potent human gonadotropin-releasing hormone receptor (hGnRH-R) antagonists is described. A major focus of this optimization was to improve the CYP3A4 inhibition liability of these uracils while maintaining their GnRH-R potency. R-4-{2-[5-(2-fluoro-3-methoxyphenyl)-3-(2-fluoro-6-[trifluoromethyl]benzyl)-4-methyl-2,6-dioxo-3,6-dihydro-2H-pyrimidin-1-yl]-1-phenylethylamino}butyric acid sodium salt, 10b (elagolix), was identified as a potent and selective hGnRH-R antagonist. Oral administration of 10b suppressed luteinizing hormone in castrated macaques. These efforts led to the identification of 10b as a clinical compound for the treatment of endometriosis.
Asunto(s)
Descubrimiento de Drogas , Hidrocarburos Fluorados/farmacología , Pirimidinas/farmacología , Receptores LHRH/antagonistas & inhibidores , Animales , Células CACO-2 , Inhibidores del Citocromo P-450 CYP3A , Evaluación Preclínica de Medicamentos , Humanos , Hidrocarburos Fluorados/química , Hidrocarburos Fluorados/metabolismo , Macaca fascicularis , Masculino , Microsomas Hepáticos/química , Microsomas Hepáticos/metabolismo , Estructura Molecular , Pirimidinas/química , Pirimidinas/metabolismo , Estereoisomerismo , Relación Estructura-Actividad , Factores de TiempoRESUMEN
The design synthesis and SAR of a series of chiral ring-constrained norepinephrine reuptake inhibitors with improved physicochemical properties is described. Typical compounds are potent (IC(50)s<10 nM), selective against the other monoamine transporters, weak CYP2D6 inhibitors (IC(50)s>1 microM) and stable to oxidation by human liver microsomes. In addition, the compounds exhibit a favorable polarity profile.
Asunto(s)
Inhibidores del Citocromo P-450 CYP2D6 , Indanos/síntesis química , Indanos/farmacología , Inhibidores de la Captación de Neurotransmisores/síntesis química , Inhibidores de la Captación de Neurotransmisores/farmacología , Norepinefrina/antagonistas & inhibidores , Clorhidrato de Atomoxetina , Técnicas Químicas Combinatorias , Diseño de Fármacos , Humanos , Indanos/química , Concentración 50 Inhibidora , Microsomas Hepáticos/metabolismo , Estructura Molecular , Inhibidores de la Captación de Neurotransmisores/química , Propilaminas/farmacología , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
The design, synthesis and SAR of a series of heterocyclic ring-constrained norepinephrine reuptake inhibitors are described. As racemates, the best compounds compare favorably with atomoxetine (IC(50)'s<10 nM) in potency at the transporter.
Asunto(s)
Proteínas de Transporte de Noradrenalina a través de la Membrana Plasmática/química , Clorhidrato de Atomoxetina , Química Farmacéutica/métodos , Inhibidores del Citocromo P-450 CYP2D6 , Citocromo P-450 CYP3A , Inhibidores del Citocromo P-450 CYP3A , Diseño de Fármacos , Humanos , Indenos/farmacología , Concentración 50 Inhibidora , Microsomas Hepáticos/enzimología , Modelos Químicos , Norepinefrina/química , Norepinefrina/metabolismo , Proteínas de Transporte de Noradrenalina a través de la Membrana Plasmática/metabolismo , Propilaminas/química , Serotonina/química , Relación Estructura-ActividadRESUMEN
The synthesis and SAR of a series of chiral heterocyclic ring-constrained norepinephrine reuptake inhibitors are described. The best compounds compare favorably with atomoxetine in potency (IC(50)s<10 nM), selectivity against the other monoamine transporters, and inhibition of CYP2D6 (IC(50)s>1 microM). In addition, the compounds are generally more stable than atomoxetine to oxidative metabolism and thus are likely to have lower clearance in humans.
Asunto(s)
Inhibidores de Captación Adrenérgica/síntesis química , Inhibidores de Captación Adrenérgica/farmacología , Química Farmacéutica/métodos , Proteínas de Transporte de Noradrenalina a través de la Membrana Plasmática/síntesis química , Proteínas de Transporte de Noradrenalina a través de la Membrana Plasmática/farmacología , Norepinefrina/química , Oxígeno/química , Inhibidores de Captación Adrenérgica/química , Clorhidrato de Atomoxetina , Citocromo P-450 CYP2D6/química , Diseño de Fármacos , Humanos , Concentración 50 Inhibidora , Modelos Químicos , Conformación Molecular , Norepinefrina/metabolismo , Propilaminas/química , Propilaminas/farmacología , Relación Estructura-Actividad , Simportadores/químicaRESUMEN
The design, synthesis, and SAR of a series of ring-constrained norepinephrine reuptake inhibitors are described. A substantially rigid inhibitor with potent functional activity at the transporter (IC(50)=8 nM) was used to develop a model for the distance and orientation of key features necessary for interaction with the norepinephrine transporter (NET).
Asunto(s)
Inhibidores de Captación Adrenérgica/síntesis química , Inhibidores de Captación Adrenérgica/farmacología , Proteínas de Transporte de Noradrenalina a través de la Membrana Plasmática/síntesis química , Proteínas de Transporte de Noradrenalina a través de la Membrana Plasmática/farmacología , Aminas/química , Clorhidrato de Atomoxetina , Sitios de Unión , Línea Celular , Química Farmacéutica/métodos , Desipramina/química , Diseño de Fármacos , Humanos , Concentración 50 Inhibidora , Modelos Químicos , Conformación Molecular , Propilaminas/química , Relación Estructura-ActividadRESUMEN
Incorporation of a carboxylic acid into a series of uracil derivatives as hGnRH-R antagonists resulted in a significant reduction of CYP3A4 inhibitory activity. Highly potent hGnRH antagonists with low CYP3A4 inhibitory liability, such as 8a and 8d, were identified. Thus, 8a had a K(i) of 2.2 nM at GnRH-R and an IC(50) of 36 microM at CYP3A4.
Asunto(s)
Inhibidores del Citocromo P-450 CYP3A , Hormona Liberadora de Gonadotropina/antagonistas & inhibidores , Receptores LHRH/antagonistas & inhibidores , Uracilo/análogos & derivados , Uracilo/síntesis química , Animales , Citocromo P-450 CYP3A , Haplorrinos , Humanos , Concentración 50 Inhibidora , Estructura Molecular , Ratas , Relación Estructura-Actividad , Uracilo/farmacocinéticaRESUMEN
The design, synthesis, and SAR of a series of retro bis-aminopyrrolidine ureas are described. Compounds from this series exhibited potent binding affinity and functional activity at MCH-R1, and good oral bioavailability in rat.
Asunto(s)
Pirrolidinas/síntesis química , Pirrolidinas/farmacología , Receptores de la Hormona Hipofisaria/antagonistas & inhibidores , Urea/análogos & derivados , Animales , Estructura Molecular , Pirrolidinas/química , Ratas , Receptores de la Hormona Hipofisaria/metabolismo , Relación Estructura-Actividad , Urea/síntesis química , Urea/química , Urea/farmacologíaRESUMEN
The design, synthesis, and SAR of a series of retro bis-aminopyrrolidine ureas are described. Compounds from this series exhibited considerable binding affinity (Ki = 1 nM) and functional activity at MCH-R1, acceptable CYP2D6 inhibition, and good rat brain exposure.
Asunto(s)
Pirrolidinas/química , Pirrolidinas/farmacología , Receptores de Somatostatina/antagonistas & inhibidores , Urea/análogos & derivados , Animales , Concentración 50 Inhibidora , Estructura Molecular , Pirrolidinas/síntesis química , Pirrolidinas/farmacocinética , Ratas , Receptores de Somatostatina/metabolismo , Relación Estructura-Actividad , Urea/síntesis química , Urea/química , Urea/farmacocinética , Urea/farmacologíaRESUMEN
A series of substituted chromones were designed, synthesized, and evaluated for their ability to bind melanin-concentrating hormone receptor 1. Compounds with subnanomolar binding affinity and 66% oral bioavailability in rats were discovered.
Asunto(s)
Química Farmacéutica/métodos , Cromonas/síntesis química , Melaninas/química , Quinolonas/síntesis química , Receptores de la Hormona Hipofisaria/antagonistas & inhibidores , Receptores de la Hormona Hipofisaria/química , Administración Oral , Animales , Cromonas/química , Diseño de Fármacos , Humanos , Microsomas Hepáticos/metabolismo , Modelos Químicos , Quinolonas/química , Ratas , Factores de TiempoRESUMEN
Uracil derivatives were designed and synthesized to avoid atropisomers observed in the 6-methyluracils as antagonists of the human GnRH receptor. Optimization at the 1- and 5-positions of the uracil resulted in potent compounds such as 24 (Ki=0.45 nM).
Asunto(s)
Receptores LHRH/antagonistas & inhibidores , Uracilo/farmacología , Humanos , Isomerismo , Estructura Molecular , Uracilo/química , Difracción de Rayos XRESUMEN
INTRODUCTION: Higher-throughput chemotaxis assays have had limited use in chemokine receptor pharmacology studies mainly because of the unavailability of optimal assay formats in addition to an incompatibility of chemotactic cell backgrounds with other pharmacological assays. Here, we developed a high-throughput 96-well chemotaxis assay for leukocytic cell lines and identified the human U937 monocytic line as an excellent cell background for both chemotaxis and the high-throughput calcium mobilization Fluorescent Imaging Plate Reader (FLIPR) assay. METHODS: Optimal chemotactic conditions were developed using the Neuroprobe MBA96 nondisposable and the Millipore MultiScreen-MIC disposable apparatuses with responses to CXC chemokine receptor (CXCR)-4 endogenously expressed on the human H9 T lymphoma line, and confirmed with Jurkat T cell and U937 monocytic cell lines. RESULTS: The U937 cell line was chosen for site-directed mutagenesis studies with CC chemokine receptor (CCR)-7 because this cell line did not endogenously express this receptor, it demonstrated a good chemotaxis index, and it showed an exceptional ability to mobilize calcium measured via FLIPR. Using the Millipore MultiScreen-MIC and FLIPR assays, alanine substitutions at K130 and Q227 caused threefold shifts in potency for the CCR7 ligand, CCL19, whereas that at K137 had no effect. DISCUSSION: Because these CCR7 mutations have previously been shown not to affect ligand binding, our results here show that these residues are specifically involved in receptor activation signals critical to chemotaxis and underscore the importance of using the U937 cell background to confirm results of chemotaxis with those of the FLIPR assay.
Asunto(s)
Quimiocinas CXC/farmacología , Quimiotaxis de Leucocito/efectos de los fármacos , Monocitos/efectos de los fármacos , Receptores de Quimiocina/fisiología , Linfocitos T/metabolismo , Secuencia de Aminoácidos , Calcio/metabolismo , Línea Celular Tumoral , Quimiocinas CXC/metabolismo , Relación Dosis-Respuesta a Droga , Citometría de Flujo , Fluorometría , Humanos , Procesamiento de Imagen Asistido por Computador , Células Jurkat , Cinética , Ligandos , Monocitos/citología , Mutación Puntual , Receptores de Quimiocina/química , Receptores de Quimiocina/genética , Reproducibilidad de los Resultados , Células U937RESUMEN
The CXC chemokine receptor 3 (CXCR3) is predominantly expressed on T helper type 1 (Th1) cells that are involved in inflammatory diseases. The three CXCR3 ligands CXCL9, CXCL10, and CXCL11 are produced at sites of inflammation and elicit migration of pathological Th1 cells. Here, we are the first to characterize the pharmacological potencies and specificity of a CXCR3 antagonist, N-1R-[3-(4-ethoxy-phenyl)-4-oxo-3,4-dihydro-pyrido[2,3-d]pyrimidin-2-yl]-ethyl-N-pyridin-3-ylmethyl-2-(4-fluoro-3-trifluoromethyl-phenyl)-acetamide (NBI-74330), from the T487 small molecule series. NBI-74330 demonstrated potent inhibition of [(125)I]CXCL10 and [(125)I]CXCL11 specific binding (K(i) of 1.5 and 3.2 nM, respectively) and of functional responses mediated by CXCR3, such as ligand-induced guanosine 5'-O-(3-[(35)S]thio)triphosphate ([(35)S]GTPgammaS) binding, calcium mobilization, and cellular chemotaxis (IC(50) of 7 to 18 nM). NBI-74330 was selective for CXCR3 because it showed no significant inhibition of chemotactic responses to other chemokines and did not inhibit radioligand binding to a panel of nonchemokine G-protein coupled receptors. There was a striking difference in potencies among the three CXCR3 ligands, with CXCL11 >> CXCL10 > CXCL9. A comparison of the rank order of K(i) values with the rank order of monocyte production levels of these three ligands revealed a precise inverse correlation, suggesting that the weaker receptor affinities of CXCL9 and CXCL10 were physiologically compensated for by an elevated expression, perhaps to maintain effectiveness of each ligand under physiological conditions.