RESUMEN
Mass spectrometry-based discovery proteomics is an essential tool for the proximal readout of cellular drug action. Here, we apply a robust proteomic workflow to rapidly profile the proteomes of five lung cancer cell lines in response to more than 50 drugs. Integration of millions of quantitative protein-drug associations substantially improved the mechanism of action (MoA) deconvolution of single compounds. For example, MoA specificity increased after removal of proteins that frequently responded to drugs and the aggregation of proteome changes across cell lines resolved compound effects on proteostasis. We leveraged these findings to demonstrate efficient target identification of chemical protein degraders. Aggregating drug response across cell lines also revealed that one-quarter of compounds modulated the abundance of one of their known protein targets. Finally, the proteomic data led us to discover that inhibition of mitochondrial function is an off-target mechanism of the MAP2K1/2 inhibitor PD184352 and that the ALK inhibitor ceritinib modulates autophagy.
Asunto(s)
Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Neoplasias Pulmonares/metabolismo , Proteómica/métodos , Antineoplásicos/farmacología , Línea Celular Tumoral , Regulación Neoplásica de la Expresión Génica/fisiología , Humanos , Espectrometría de Masas , ProteomaRESUMEN
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
RESUMEN
The transformation of an aryloxybutanoic acid ultra high-throughput screening (uHTS) hit into a potent and selective series of G-protein coupled receptor 120 (GPR120) agonists is reported. uHTS hit 1 demonstrated an excellent rodent pharmacokinetic profile and selectivity over the related fatty acid receptor GPR40, but only modest GPR120 potency. Optimization of the "left-hand" aryl group led to compound 6, which demonstrated a GPR120 mechanism-based pharmacodynamic effect in a mouse oral glucose tolerance test (oGTT). Further optimization gave rise to the benzofuran propanoic acid series (exemplified by compound 37), which demonstrated acute mechanism-based pharmacodynamic effects. The combination of in vivo efficacy and attractive rodent pharmacodynamic profiles suggests compounds generated from this series may afford attractive candidates for the treatment of Type 2 diabetes.
Asunto(s)
Benzofuranos/química , Benzofuranos/farmacología , Propionatos/química , Propionatos/farmacología , Receptores Acoplados a Proteínas G/agonistas , Animales , Benzofuranos/sangre , Glucemia/análisis , Glucemia/metabolismo , Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Diabetes Mellitus Tipo 2/metabolismo , Evaluación Preclínica de Medicamentos , Ensayos Analíticos de Alto Rendimiento , Humanos , Hipoglucemiantes/sangre , Hipoglucemiantes/química , Hipoglucemiantes/farmacología , Ratones , Propionatos/sangre , Receptores Acoplados a Proteínas G/metabolismoRESUMEN
A novel, potent series of glucagon receptor antagonists (GRAs) was discovered. These indazole- and indole-based compounds were designed on an earlier pyrazole-based GRA lead MK-0893. Structure-activity relationship (SAR) studies were focused on the C3 and C6 positions of the indazole core, as well as the benzylic position on the N-1 of indazole. Multiple potent GRAs were identified with excellent in vitro profiles and good pharmacokinetics in rat. Among them, GRA 16d was found to be orally active in blunting glucagon induced glucose excursion in an acute glucagon challenge model in glucagon receptor humanized (hGCGR) mice at 1, 3 and 10mg/kg (mpk), and significantly lowered acute glucose levels in hGCGR ob/ob mice at 3 mpk dose.
Asunto(s)
Indazoles/química , Indazoles/farmacología , Indoles/química , Indoles/farmacología , Receptores de Glucagón/antagonistas & inhibidores , Animales , Células CHO , Cricetulus , Relación Dosis-Respuesta a Droga , Humanos , Ratones , Ratones Obesos , Estructura Molecular , Ratas , Relación Estructura-ActividadRESUMEN
Novel prolylcarboxypeptidase (PrCP) inhibitors with nanomolar IC(50) values were prepared by replacing the previously described dichlorobenzimidazole-substituted pyrrolidine amides with a variety of substituted benzylamine amides. In contrast to prior series, the compounds demonstrated minimal inhibition shift in whole serum and minimal recognition by P-glycoprotein (P-gp) efflux transporters. The compounds were also cell permeable and demonstrated in vivo brain exposure. The in vivo effect of compound (S)-6e on weight loss in an established diet-induced obesity (eDIO) mouse model was studied.
Asunto(s)
Bencimidazoles/farmacología , Encéfalo/metabolismo , Carboxipeptidasas/antagonistas & inhibidores , Subfamilia B de Transportador de Casetes de Unión a ATP/genética , Subfamilia B de Transportador de Casetes de Unión a ATP/metabolismo , Amidas/química , Animales , Transporte Biológico , Peso Corporal , Encéfalo/efectos de los fármacos , Modelos Animales de Enfermedad , Humanos , Concentración 50 Inhibidora , Ratones , Modelos Químicos , Obesidad/tratamiento farmacológico , Pirrolidinas/química , Factores de TiempoRESUMEN
Efforts were dedicated to develop potent and brain penetrant prolylcarboxypeptidase (PrCP) inhibitors by replacing the amide group of original leads 1 and 2 with heterocycles. Aminopyrimidines including compound 32a were identified to display good PrCP inhibitory activity (32a, IC(50)=43 nM) and impressive ability to penetrate brain in mice (brain/plasma ratio: 1.4).
Asunto(s)
Aminas/síntesis química , Encéfalo/efectos de los fármacos , Carboxipeptidasas/antagonistas & inhibidores , Descubrimiento de Drogas , Inhibidores Enzimáticos , Compuestos Heterocíclicos/síntesis química , Compuestos Heterocíclicos/farmacología , Amidas/química , Aminas/química , Aminas/farmacología , Animales , Encéfalo/enzimología , Activación Enzimática/efectos de los fármacos , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/farmacología , Compuestos Heterocíclicos/química , Concentración 50 Inhibidora , Ratones , Estructura Molecular , Relación Estructura-ActividadRESUMEN
Efforts to modify the central proline portion of lead compound 4 lead to the discovery of novel prolylcarboxypeptidase (PrCP) inhibitors. Especially, replacement with alanine afforded compound 19 displaying more potent human and mouse PrCP inhibitory activity than 4 and an overall comparable profile.
Asunto(s)
Alanina/química , Carboxipeptidasas/antagonistas & inhibidores , Descubrimiento de Drogas , Alanina/farmacología , Animales , Activación Enzimática/efectos de los fármacos , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Humanos , Concentración 50 Inhibidora , Ratones , Estructura Molecular , Relación Estructura-ActividadRESUMEN
Stereochemically and structurally complex cyclic dinucleotide-based stimulator of interferon genes (STING) agonists were designed and synthesized to access a previously unexplored chemical space. The assessment of biochemical affinity and cellular potency, along with computational, structural, and biophysical characterization, was applied to influence the design and optimization of novel STING agonists, resulting in the discovery of MK-1454 as a molecule with appropriate properties for clinical development. When administered intratumorally to immune-competent mice-bearing syngeneic tumors, MK-1454 exhibited robust tumor cytokine upregulation and effective antitumor activity. Tumor shrinkage in mouse models that are intrinsically resistant to single-agent therapy was further enhanced when treating the animals with MK-1454 in combination with a fully murinized antimouse PD-1 antibody, mDX400. These data support the development of STING agonists in combination with pembrolizumab (humanized anti-PD-1 antibody) for patients with tumors that are partially responsive or nonresponsive to single-agent anti-PD-1 therapy.
Asunto(s)
Proteínas de la Membrana , Neoplasias , Animales , Citocinas , Humanos , Inmunoterapia/métodos , Interferones , Ratones , Neoplasias/tratamiento farmacológicoRESUMEN
Amino-anthranilic acid derivatives have been identified as a new class of low serum shifted, high affinity full agonists of the human orphan G-protein-coupled receptor GPR109a with improved ADME properties.
Asunto(s)
Descubrimiento de Drogas , Receptores Acoplados a Proteínas G/agonistas , Animales , Flúor/química , Humanos , Concentración 50 Inhibidora , Ratones , Estructura Molecular , Niacina/síntesis química , Niacina/química , Niacina/farmacología , Piridinas/síntesis química , Piridinas/química , Ratas , Ratas Sprague-Dawley , Receptores NicotínicosRESUMEN
A series of benzimidazole pyrrolidinyl amides containing a piperidinyl group were discovered as novel prolylcarboxypeptidase (PrCP) inhibitors. Low-nanomolar IC(50)'s were achieved for several analogs, of which compound 9b displayed modest ex vivo target engagement in eDIO mouse plasma. Compound 9b was also studied in vivo for its effect on weight loss and food intake in an eDIO mouse model and the results will be discussed.
Asunto(s)
Amidas , Bencimidazoles , Carboxipeptidasas/antagonistas & inhibidores , Descubrimiento de Drogas , Inhibidores Enzimáticos , Pirrolidinas , Amidas/química , Amidas/farmacología , Animales , Bencimidazoles/química , Bencimidazoles/farmacología , Modelos Animales de Enfermedad , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Humanos , Concentración 50 Inhibidora , Ratones , Estructura Molecular , Pirrolidinas/química , Pirrolidinas/farmacología , Relación Estructura-ActividadRESUMEN
A novel class of 1,3,5-pyrazoles has been discovered as potent human glucagon receptor antagonists. Notably, compound 26 is orally bioavailable in several preclinical species and shows selectivity towards cardiac ion channels, other family B receptors such hGIP and hGLP1, and a large panel of enzymes and additional receptors. When dosed orally, compound 26 is efficacious in suppressing glucagon induced plasma glucose excursion in rhesus monkey and transgenic murine pharmacodynamic models at 1 and 10 mpk, respectively.
Asunto(s)
Pirazoles/química , Receptores de Glucagón/antagonistas & inhibidores , Administración Oral , Animales , Glucemia/metabolismo , Perros , Evaluación Preclínica de Medicamentos , Humanos , Macaca mulatta , Ratones , Ratones Transgénicos , Pirazoles/síntesis química , Pirazoles/farmacocinética , Ratas , Receptores de Glucagón/metabolismo , Relación Estructura-ActividadRESUMEN
A series of pyrazolyl propionyl cyclohexenamides were discovered as full agonists for the high affinity niacin receptor GPR109A. The structure-activity relationship (SAR) studies were aimed to improve activity on GPR109A, reduce Cytochrome P450 2C8 (CYP2C8) and Cytochrome P450 2C9 (CYP2C9) inhibition, reduce serum shift and improve pharmacokinetic (PK) profiles.
Asunto(s)
Amidas/química , Ciclohexenos/farmacología , Receptores Acoplados a Proteínas G/agonistas , Animales , Ciclohexenos/química , Ciclohexenos/farmacocinética , Ratones , Ratas , Receptores Nicotínicos , Relación Estructura-ActividadRESUMEN
5-Alkyl and aryl-pyrazole-acids have been identified as a new class of selective, small-molecule, agonists of the human orphan G-protein-coupled receptor GPR109a, a high affinity receptor for the HDL-raising drug nicotinic acid.
Asunto(s)
Pirazoles/química , Receptores Acoplados a Proteínas G/agonistas , Animales , Ácidos Grasos/metabolismo , Humanos , Ratones , Niacina/química , Pirazoles/síntesis química , Pirazoles/farmacocinética , Receptores Acoplados a Proteínas G/metabolismo , Receptores Nicotínicos/metabolismoRESUMEN
Proprotein convertase substilisin-like/kexin type 9 (PCSK9) is a serine protease involved in a protein-protein interaction with the low-density lipoprotein (LDL) receptor that has both human genetic and clinical validation. Blocking this protein-protein interaction prevents LDL receptor degradation and thereby decreases LDL cholesterol levels. Our pursuit of small-molecule direct binders for this difficult to drug PPI target utilized affinity selection/mass spectrometry, which identified one confirmed hit compound. An X-ray crystal structure revealed that this compound was binding in an unprecedented allosteric pocket located between the catalytic and C-terminal domain. Optimization of this initial hit, using two distinct strategies, led to compounds with high binding affinity to PCSK9. Direct target engagement was demonstrated in the cell lysate with a cellular thermal shift assay. Finally, ligand-induced protein degradation was shown with a proteasome recruiting tag attached to the high-affinity allosteric ligand for PCSK9.
Asunto(s)
Descubrimiento de Drogas , Evaluación Preclínica de Medicamentos , Proproteína Convertasa 9/metabolismo , Proteolisis/efectos de los fármacos , Inhibidores de Serina Proteinasa/farmacología , Bibliotecas de Moléculas Pequeñas/farmacología , Humanos , Ligandos , Modelos Moleculares , Estructura Molecular , Inhibidores de Serina Proteinasa/química , Bibliotecas de Moléculas Pequeñas/químicaRESUMEN
Distinct from previously reported urea and amide inhibitors of soluble epoxide hydrolase (sEH), a novel class of inhibitors were rationally designed based on the X-ray structure of this enzyme and known amide inhibitors. The structure-activity relationship (SAR) study was focused on improving the sEH inhibitory activity. Aminobenzisoxazoles emerged to be the optimal series, of which a potent human sEH inhibitor 7t was identified with a good pharmacokinetics (PK) profile. The strategy of employing aminoheterocycles as amide replacements may represent a general approach to develop mimics of known hydrolase or protease inhibitors containing an amide moiety.
Asunto(s)
Amidas/química , Compuestos de Anilina/química , Inhibidores Enzimáticos/química , Epóxido Hidrolasas/antagonistas & inhibidores , Compuestos Heterocíclicos con 2 Anillos/química , Isoxazoles/química , Compuestos de Anilina/síntesis química , Compuestos de Anilina/farmacocinética , Animales , Sitios de Unión , Simulación por Computador , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/farmacocinética , Epóxido Hidrolasas/metabolismo , Compuestos Heterocíclicos con 2 Anillos/síntesis química , Compuestos Heterocíclicos con 2 Anillos/farmacocinética , Humanos , Isoxazoles/síntesis química , Isoxazoles/farmacocinética , Unión Proteica , Ratas , Relación Estructura-ActividadRESUMEN
Platensimycin (1) displays antibacterial activity due to its inhibition of the elongation condensing enzyme (FabF), a novel mode of action that could potentially lead to a breakthrough in developing a new generation of antibiotics. The medicinal chemistry efforts were focused on the modification of the enone moiety of platensimycin and several analogs showed significant activity against FabF and possess antibacterial activity.
Asunto(s)
3-Oxoacil-(Proteína Transportadora de Acil) Sintasa/antagonistas & inhibidores , Adamantano/síntesis química , Aminobenzoatos/síntesis química , Anilidas/síntesis química , Antibacterianos/síntesis química , Proteínas Bacterianas/antagonistas & inhibidores , Química Farmacéutica/métodos , 3-Oxoacil-(Proteína Transportadora de Acil) Sintasa/química , Adamantano/farmacología , Aminobenzoatos/farmacología , Anilidas/farmacología , Antibacterianos/farmacología , Proteínas Bacterianas/química , Cristalografía por Rayos X/métodos , Diseño de Fármacos , Farmacorresistencia Microbiana , Enterococcus faecalis/metabolismo , Concentración 50 Inhibidora , Meticilina/farmacología , Pruebas de Sensibilidad Microbiana , Modelos Químicos , Estructura Molecular , Streptomyces/metabolismo , Relación Estructura-ActividadRESUMEN
5-Alkyl and aryl-pyrazole-tetrazoles have been identified as a new class of selective, small-molecule, agonists of the human G-protein-coupled receptor GPR109a, a high affinity receptor for the HDL-raising drug nicotinic acid.
Asunto(s)
Pirazoles/química , Receptores Acoplados a Proteínas G/agonistas , Tetrazoles/química , Animales , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Agonistas Nicotínicos/química , Agonistas Nicotínicos/farmacología , Pirazoles/farmacología , Receptores Acoplados a Proteínas G/deficiencia , Receptores Acoplados a Proteínas G/fisiología , Receptores Nicotínicos/deficiencia , Receptores Nicotínicos/fisiología , Tetrazoles/farmacología , Vasodilatación/efectos de los fármacos , Vasodilatación/fisiologíaRESUMEN
3,3-Disubstituted piperidine-derived trisubstituted urea entA-2b was discovered as a highly potent and selective soluble epoxide hydrolase (sEH) inhibitor. Despite the good compound oral exposure, excellent sEH inhibition in whole blood, and remarkable selectivity, compound entA-2b failed to lower blood pressure acutely in spontaneously hypertensive rats (SHRs). This observation further challenges the premise that sEH inhibition can provide a viable approach to the treatment of hypertensive patients.
Asunto(s)
Epóxido Hidrolasas/antagonistas & inhibidores , Epóxido Hidrolasas/metabolismo , Hipertensión/tratamiento farmacológico , Piperidinas/química , Urea/análogos & derivados , Urea/farmacología , Ácido 8,11,14-Eicosatrienoico/análogos & derivados , Ácido 8,11,14-Eicosatrienoico/sangre , Ácido 8,11,14-Eicosatrienoico/metabolismo , Animales , Presión Sanguínea/efectos de los fármacos , Humanos , Modelos Moleculares , Unión Proteica , Ratas , Ratas Endogámicas SHR , Relación Estructura-Actividad , Urea/uso terapéuticoRESUMEN
Spirocyclic secondary amine-derived trisubstituted ureas were identified as highly potent, bioavailable and selective soluble epoxide hydrolase (sEH) inhibitors. Despite good oral exposure and excellent ex vivo target engagement in blood, one such compound, rac-1a, failed to lower blood pressure acutely in spontaneously hypertensive rats (SHRs). This study posed the question as to whether sEH inhibition provides a robust mechanism leading to a significant antihypertensive effect.