RESUMO
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.
Assuntos
Proteases 3C de Coronavírus/química , Proteases Semelhantes à Papaína de Coronavírus/química , Desenvolvimento de Medicamentos/métodos , Descoberta de Drogas/métodos , Ensaios de Triagem em Larga Escala/métodos , Histona-Lisina N-Metiltransferase/química , Animais , Sobrevivência Celular , Chlorocebus aethiops , Química Computacional , Cristalografia por Raios X , Bases de Dados de Proteínas , Desenho de Fármacos , Células HEK293 , Humanos , Ligação de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Ligantes , Ligação Proteica , Receptores Acoplados a Proteínas G/química , SARS-CoV-2/química , SARS-CoV-2/genética , Bibliotecas de Moléculas Pequenas , Células VeroRESUMO
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).
Assuntos
Inibidores da Captação Adrenérgica/síntese química , Inibidores da Captação Adrenérgica/farmacologia , Proteínas da Membrana Plasmática de Transporte de Norepinefrina/síntese química , Proteínas da Membrana Plasmática de Transporte de Norepinefrina/farmacologia , Aminas/química , Cloridrato de Atomoxetina , Sítios de Ligação , Linhagem Celular , Química Farmacêutica/métodos , Desipramina/química , Desenho de Fármacos , Humanos , Concentração Inibidora 50 , Modelos Químicos , Conformação Molecular , Propilaminas/química , Relação Estrutura-AtividadeRESUMO
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.
Assuntos
Proteínas da Membrana Plasmática de Transporte de Norepinefrina/química , Cloridrato de Atomoxetina , Química Farmacêutica/métodos , Inibidores do Citocromo P-450 CYP2D6 , Citocromo P-450 CYP3A , Inibidores do Citocromo P-450 CYP3A , Desenho de Fármacos , Humanos , Indenos/farmacologia , Concentração Inibidora 50 , Microssomos Hepáticos/enzimologia , Modelos Químicos , Norepinefrina/química , Norepinefrina/metabolismo , Proteínas da Membrana Plasmática de Transporte de Norepinefrina/metabolismo , Propilaminas/química , Serotonina/química , Relação Estrutura-AtividadeRESUMO
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.
Assuntos
Inibidores da Captação Adrenérgica/síntese química , Inibidores da Captação Adrenérgica/farmacologia , Química Farmacêutica/métodos , Proteínas da Membrana Plasmática de Transporte de Norepinefrina/síntese química , Proteínas da Membrana Plasmática de Transporte de Norepinefrina/farmacologia , Norepinefrina/química , Oxigênio/química , Inibidores da Captação Adrenérgica/química , Cloridrato de Atomoxetina , Citocromo P-450 CYP2D6/química , Desenho de Fármacos , Humanos , Concentração Inibidora 50 , Modelos Químicos , Conformação Molecular , Norepinefrina/metabolismo , Propilaminas/química , Propilaminas/farmacologia , Relação Estrutura-Atividade , Simportadores/químicaRESUMO
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.
Assuntos
Inibidores do Citocromo P-450 CYP2D6 , Indanos/síntese química , Indanos/farmacologia , Inibidores da Captação de Neurotransmissores/síntese química , Inibidores da Captação de Neurotransmissores/farmacologia , Norepinefrina/antagonistas & inibidores , Cloridrato de Atomoxetina , Técnicas de Química Combinatória , Desenho de Fármacos , Humanos , Indanos/química , Concentração Inibidora 50 , Microssomos Hepáticos/metabolismo , Estrutura Molecular , Inibidores da Captação de Neurotransmissores/química , Propilaminas/farmacologia , Estereoisomerismo , Relação Estrutura-AtividadeRESUMO
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.
Assuntos
Inibidores do Citocromo P-450 CYP3A , Hormônio Liberador de Gonadotropina/antagonistas & inibidores , Receptores LHRH/antagonistas & inibidores , Uracila/análogos & derivados , Uracila/síntese química , Animais , Citocromo P-450 CYP3A , Haplorrinos , Humanos , Concentração Inibidora 50 , Estrutura Molecular , Ratos , Relação Estrutura-Atividade , Uracila/farmacocinéticaRESUMO
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.
Assuntos
Quimiocinas CXC/farmacologia , Quimiotaxia de Leucócito/efeitos dos fármacos , Monócitos/efeitos dos fármacos , Receptores de Quimiocinas/fisiologia , Linfócitos T/metabolismo , Sequência de Aminoácidos , Cálcio/metabolismo , Linhagem Celular Tumoral , Quimiocinas CXC/metabolismo , Relação Dose-Resposta a Droga , Citometria de Fluxo , Fluorometria , Humanos , Processamento de Imagem Assistida por Computador , Células Jurkat , Cinética , Ligantes , Monócitos/citologia , Mutação Puntual , Receptores de Quimiocinas/química , Receptores de Quimiocinas/genética , Reprodutibilidade dos Testes , Células U937RESUMO
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.
Assuntos
Descoberta de Drogas , Hidrocarbonetos Fluorados/farmacologia , Pirimidinas/farmacologia , Receptores LHRH/antagonistas & inibidores , Animais , Células CACO-2 , Inibidores do Citocromo P-450 CYP3A , Avaliação Pré-Clínica de Medicamentos , Humanos , Hidrocarbonetos Fluorados/química , Hidrocarbonetos Fluorados/metabolismo , Macaca fascicularis , Masculino , Microssomos Hepáticos/química , Microssomos Hepáticos/metabolismo , Estrutura Molecular , Pirimidinas/química , Pirimidinas/metabolismo , Estereoisomerismo , Relação Estrutura-Atividade , Fatores de TempoRESUMO
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.
Assuntos
Química Farmacêutica/métodos , Cromonas/síntese química , Melaninas/química , Quinolonas/síntese química , Receptores do Hormônio Hipofisário/antagonistas & inibidores , Receptores do Hormônio Hipofisário/química , Administração Oral , Animais , Cromonas/química , Desenho de Fármacos , Humanos , Microssomos Hepáticos/metabolismo , Modelos Químicos , Quinolonas/química , Ratos , Fatores de TempoRESUMO
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.
Assuntos
Pirrolidinas/síntese química , Pirrolidinas/farmacologia , Receptores do Hormônio Hipofisário/antagonistas & inibidores , Ureia/análogos & derivados , Animais , Estrutura Molecular , Pirrolidinas/química , Ratos , Receptores do Hormônio Hipofisário/metabolismo , Relação Estrutura-Atividade , Ureia/síntese química , Ureia/química , Ureia/farmacologiaRESUMO
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.
Assuntos
Pirrolidinas/química , Pirrolidinas/farmacologia , Receptores de Somatostatina/antagonistas & inibidores , Ureia/análogos & derivados , Animais , Concentração Inibidora 50 , Estrutura Molecular , Pirrolidinas/síntese química , Pirrolidinas/farmacocinética , Ratos , Receptores de Somatostatina/metabolismo , Relação Estrutura-Atividade , Ureia/síntese química , Ureia/química , Ureia/farmacocinética , Ureia/farmacologiaRESUMO
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).
Assuntos
Receptores LHRH/antagonistas & inibidores , Uracila/farmacologia , Humanos , Isomerismo , Estrutura Molecular , Uracila/química , Difração de Raios XRESUMO
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.