RESUMO
A series of non-steroidal GPBAR1 (TGR5) agonists was developed from a hit in a high-throughput screening campaign. Lead identification efforts produced biphenyl-4-carboxylic acid derivative (R)-22, which displayed a robust secretion of PYY after oral administration in a degree that can be correlated with the unbound plasma concentration. Further optimisation work focusing on reduction of the lipophilicity provided the 1-phenylpiperidine-4-carboxylic acid derivative (R)-29 (RO5527239), which showed an improved secretion of PYY and GLP-1, translating into a significant reduction of postprandial blood glucose excursion in an oral glucose tolerance test in DIO mice.
Assuntos
Glicemia/efeitos dos fármacos , Descoberta de Drogas , Oximas/síntese química , Propano/análogos & derivados , Receptores Acoplados a Proteínas G/agonistas , Administração Oral , Animais , Concentração Inibidora 50 , Camundongos , Estrutura Molecular , Oximas/química , Oximas/farmacologia , Propano/sangue , Propano/síntese química , Propano/química , Propano/farmacologiaRESUMO
Pyrido pyrimidinones are selective agonists of the human high affinity niacin receptor GPR109A (HM74A). They show no activity on the highly homologous low affinity receptor GPR109B (HM74). Starting from a high throughput screening hit the in vitro activity of the pyrido pyrimidinones was significantly improved providing lead compounds suitable for further optimization.
Assuntos
Niacina/metabolismo , Pirimidinonas/química , Pirimidinonas/farmacologia , Receptores Acoplados a Proteínas G/agonistas , Receptores Acoplados a Proteínas G/metabolismo , Receptores Nicotínicos/metabolismo , Animais , Microssomos Hepáticos/metabolismo , Pirimidinonas/administração & dosagem , Pirimidinonas/metabolismo , Ratos , Ratos Wistar , Relação Estrutura-AtividadeRESUMO
In higher organisms the formation of the steroid scaffold is catalysed exclusively by the membrane-bound oxidosqualene cyclase (OSC; lanosterol synthase). In a highly selective cyclization reaction OSC forms lanosterol with seven chiral centres starting from the linear substrate 2,3-oxidosqualene. Valuable data on the mechanism of the complex cyclization cascade have been collected during the past 50 years using suicide inhibitors, mutagenesis studies and homology modelling. Nevertheless it is still not fully understood how the enzyme catalyses the reaction. Because of the decisive role of OSC in cholesterol biosynthesis it represents a target for the discovery of novel anticholesteraemic drugs that could complement the widely used statins. Here we present two crystal structures of the human membrane protein OSC: the target protein with an inhibitor that showed cholesterol lowering in vivo opens the way for the structure-based design of new OSC inhibitors. The complex with the reaction product lanosterol gives a clear picture of the way in which the enzyme achieves product specificity in this highly exothermic cyclization reaction.
Assuntos
Transferases Intramoleculares/química , Transferases Intramoleculares/metabolismo , Lanosterol/metabolismo , Esqualeno/análogos & derivados , Anticolesterolemiantes/química , Anticolesterolemiantes/farmacologia , Benzofenonas/química , Benzofenonas/farmacologia , Catálise , Cristalografia por Raios X , Ciclização , Desenho de Fármacos , Humanos , Transferases Intramoleculares/antagonistas & inibidores , Lanosterol/química , Modelos Moleculares , Esqualeno/metabolismo , Relação Estrutura-AtividadeRESUMO
A series of 25 compounds, some of which previously were described as inhibitors of human liver microsomal oxidosqualene cyclase (OSC), were tested as inhibitors of Saccharomyces cerevisiae, Trypanosoma cruzi, Pneumocystis carinii and Arabidopsis thaliana OSCs expressed in an OSC-defective strain of S. cerevisiae. The screening identified three derivatives particularly promising for the development of novel anti-Trypanosoma agents and eight derivatives for the development of novel anti-Pneumocystis agents.
Assuntos
Antiparasitários/síntese química , Antiparasitários/farmacologia , Arabidopsis/enzimologia , Química Farmacêutica/métodos , Transferases Intramoleculares/química , Pneumocystis carinii/enzimologia , Saccharomyces cerevisiae/enzimologia , Trypanosoma cruzi/enzimologia , Animais , Cromatografia em Camada Fina , Desenho de Fármacos , Humanos , Transferases Intramoleculares/metabolismo , Microssomos Hepáticos/metabolismo , Modelos Químicos , Esteróis/químicaRESUMO
A series of tetrahydro-cyclopenta[b]indoles modulating the activity of the liver-X-receptor (LXR) were derived from a high throughput screening hit. The potency and selectivity for LXRbeta versus LXRalpha was improved. One compound, administered to wild-type mice modestly increased plasma HDL-cholesterol with no change in plasma triglycerides (TG) and reduced effects on liver TG content compared to T0901317. This novel series of LXR agonists shows promise to improve therapeutic efficacy with reduced potential to increase TG.
Assuntos
Química Farmacêutica/métodos , Proteínas de Ligação a DNA/química , Indóis/síntese química , Receptores Citoplasmáticos e Nucleares/química , Animais , HDL-Colesterol/metabolismo , Desenho de Fármacos , Hidrocarbonetos Fluorados/farmacologia , Indóis/farmacologia , Concentração Inibidora 50 , Receptores X do Fígado , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Modelos Químicos , Receptores Nucleares Órfãos , Sulfonamidas/farmacologia , Ativação Transcricional , Triglicerídeos/metabolismoRESUMO
OBJECTIVE: Inhibition of 2,3-oxidosqualene:lanosterol cyclase (OSC), an enzyme in the cholesterol synthesis pathway, has the unique ability to inhibit cholesterol synthesis while simultaneously enhancing oxysterol synthesis. Our objectives were to determine, in vivo, if a novel OSC inhibitor reduced low-density lipoprotein (LDL) cholesterol and to define the mechanism(s) involved. METHODS AND RESULTS: Miniature pigs received the OSC inhibitor RO0717625 or placebo and a diet containing fat (34% of energy) and 400 mg per day of cholesterol. Treatment decreased plasma total cholesterol (-20%) and LDL cholesterol (-29%). Apolipoprotein B (apoB) kinetic parameters were determined. Very low-density lipoprotein (VLDL) apoB pool size decreased 22% because of inhibition of VLDL production (-43%). LDL apoB pool size decreased 22% because of a 1.5-fold increase in fractional catabolic rate (FCR). The increased FCR was associated with a 2-fold increase in hepatic LDL receptor mRNA. Hepatic total and microsomal cholesterol were reduced by 16% and 27%, respectively. Plasma lathosterol concentrations decreased 57%, reflecting inhibition of hepatic cholesterol synthesis. Treatment reduced plasma plant sterols and decreased postprandial cholesterol transport in chylomicrons. CONCLUSIONS: A novel OSC inhibitor, RO0717625, decreased VLDL and LDL apoB100 through decreased VLDL production and enhanced LDL clearance. Thus, OSC represents a potential therapeutic target for dyslipidemia.
Assuntos
Apolipoproteínas B/metabolismo , Colesterol/sangue , Inibidores Enzimáticos/farmacologia , Transferases Intramoleculares/antagonistas & inibidores , Fígado/metabolismo , Transportadores de Cassetes de Ligação de ATP/genética , Animais , Apolipoproteína B-100 , Apolipoproteínas B/biossíntese , Colesterol/biossíntese , HDL-Colesterol/biossíntese , HDL-Colesterol/sangue , LDL-Colesterol/biossíntese , LDL-Colesterol/sangue , VLDL-Colesterol/biossíntese , VLDL-Colesterol/sangue , Hidroximetilglutaril-CoA Redutases/genética , Hidroximetilglutaril-CoA Redutases/metabolismo , Fitosteróis/sangue , RNA Mensageiro/análise , Receptores de LDL/genética , Suínos , Porco MiniaturaRESUMO
Endothelin-1 (ET-1) is mitogenic and/or antiapoptotic in human cancers, and antagonists to ET-1 receptors are under evaluation for cancer treatment. Inhibition of ET-1 activation by the endothelin-converting enzymes 1(a)(-)(d) (ECE-1(a)(-)(d); EC 3.4.24.71) represents another approach to block the ET-1 effect in cancer. To evaluate this potential, we synthesized and characterized a series of low nanomolar nonpeptidic thiol-containing ECE-1 inhibitors, and evaluated their effect, as well as the effect of inhibitors for the related metalloproteases neprilysin (NEP; EC 3.4.24.11) and angiotensin-converting enzyme (ACE; EC 3.4.15.1), on human glioblastoma cell growth. Only ECE-1 inhibitors inhibited DNA synthesis by human glioblastoma cells. Exogenous addition of ET-1 or bigET-1 to glioblastoma cells did not counterbalance the growth inhibition elicited by ECE-1 inhibitors, suggesting that ECE-1 inhibitors block the proliferation of human glioblastoma cells most likely via a mechanism not involving extracellular production of ET-1. This class of molecules may thus represent novel therapeutic agents for the potential treatment of human cancer.
Assuntos
Antineoplásicos/síntese química , Ácido Aspártico Endopeptidases/antagonistas & inibidores , Compostos de Sulfidrila/síntese química , Antineoplásicos/química , Antineoplásicos/farmacologia , Carbamatos/síntese química , Carbamatos/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Neoplasias do Sistema Nervoso Central , Ensaios de Seleção de Medicamentos Antitumorais , Endotelina-1/farmacologia , Enzimas Conversoras de Endotelina , Glioblastoma , Humanos , Hidrazinas/síntese química , Hidrazinas/química , Metaloendopeptidases , Prolina/análogos & derivados , Prolina/síntese química , Prolina/química , Pirimidinas/síntese química , Pirimidinas/química , Pirrolidinas/síntese química , Pirrolidinas/química , Relação Estrutura-Atividade , Compostos de Sulfidrila/química , Compostos de Sulfidrila/farmacologiaRESUMO
The binding structures of 11 human oxidosqualene cyclase inhibitors designed as cholesterol-lowering agents were determined for the squalene-hopene cyclase from Alicyclobacillus acidocaldarius, which is the only structurally known homologue of the human enzyme. The complexes were produced by cocrystallization, and the structures were elucidated by X-ray diffraction analyses. All inhibitors were bound in the large active center cavity. The detailed binding structures are presented and discussed in the light of the IC50 values of these 11 as well as 17 other inhibitors. They provide a consistent picture for the inhibition of the bacterial enzyme and can be used to adjust and improve homology models of the human enzyme. The detailed active center structures of the two enzymes are too different to show an IC50 correlation.
Assuntos
Anticolesterolemiantes/química , Inibidores Enzimáticos/química , Transferases Intramoleculares/antagonistas & inibidores , Aminas/química , Anticolesterolemiantes/farmacologia , Bacillaceae/química , Derivados de Benzeno/química , Benzofenonas/química , Benzofenonas/farmacologia , Sítios de Ligação , Cristalografia por Raios X , Ciclopropanos/química , Inibidores Enzimáticos/farmacologia , Humanos , Técnicas In Vitro , Transferases Intramoleculares/química , Microssomos Hepáticos/efeitos dos fármacos , Microssomos Hepáticos/metabolismo , Modelos Moleculares , Ligação Proteica , Relação Estrutura-AtividadeRESUMO
New orally active non-terpenoic inhibitors of human 2,3-oxidosqualene cyclase (hOSC) are reported. The starting point for the optimization process was a set of compounds derived from a fungicide project, which in addition to showing high affinity for OSC from Candida albicans showed also high affinity for human OSC. Common structural elements of these inhibitors are an amine residue and an electrophilic carbonyl C atom embedded in a benzophenone system, which are at a distance of about 10.7 A. Considering that the keto moiety is in a potentially labile position, modifications of the substitution pattern at the benzophenone as well as annelated heteroaryl systems were explored. Our approach combined testing of the compounds first for increased binding affinity and for increased stability in vitro. Most promising compounds were then evaluated for their efficacy in lowering plasma total cholesterol (TC) and plasma low-density lipoprotein cholesterol (LDL-C) in hyperlipidemic hamsters. In this respect, the most promising compounds are the benzophenone derivative 1.fumarate and the benzo[d]isothiazol 24.fumarate, which lowered TC by 40% and 33%, respectively.
Assuntos
Alilamina/síntese química , Anticolesterolemiantes/síntese química , Benzofenonas/síntese química , Transferases Intramoleculares/antagonistas & inibidores , Tiazóis/síntese química , Administração Oral , Alilamina/análogos & derivados , Alilamina/química , Alilamina/farmacologia , Animais , Anticolesterolemiantes/química , Anticolesterolemiantes/farmacologia , Benzofenonas/química , Benzofenonas/farmacologia , Candida albicans/enzimologia , Colesterol/sangue , LDL-Colesterol/sangue , Cricetinae , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Humanos , Técnicas In Vitro , Ratos , Relação Estrutura-Atividade , Tiazóis/química , Tiazóis/farmacologiaRESUMO
Potency with potential: 2-Phenoxy-nicotinamides were identified as potent agonists at the GPBAR1 receptor, a target in the treatment of obesity, type 2 diabetes and metabolic syndrome. Extensive structure-activity relationship studies supported by homology modeling and docking resulted in the identification of optimized GPBAR1 agonists, potent against both human and mouse receptors, endowed with favorable physicochemical properties and good metabolic stability.
Assuntos
Niacinamida/química , Receptores Acoplados a Proteínas G/agonistas , Sítios de Ligação , Humanos , Simulação de Acoplamento Molecular , Niacinamida/metabolismo , Ligação Proteica , Estrutura Terciária de Proteína , Quinolinas/química , Receptores Acoplados a Proteínas G/metabolismo , Relação Estrutura-AtividadeRESUMO
Ten oxidosqualene cyclase inhibitors with high efficacy as cholesterol-lowering agents and of different chemical structure classes were evaluated as potential anticancer agents against human cancer cells from various tissue origins and nontumoral human-brain-derived endothelial cells. Inhibition of cancer cell growth was demonstrated at micromolar concentrations, comparable to the concentrations of statins necessary for antitumor effect. Human glioblastoma cells were among the most sensitive cells. These compounds were also able to decrease the proliferation of angiogenic brain-derived endothelial cells, as a model of tumor-induced neovasculation. Additive effects in human glioblastoma cells were also demonstrated for oxidosqualene cyclase inhibitors in combination with atorvastatin while maintaining selectivity against endothelial cells. Thus, not only statins targeting the 3-hydroxy-3-methylglutaryl coenzyme A reductase but also inhibitors of oxidosqualene cyclase decrease tumor growth, suggesting new therapeutic opportunities of combined anti-cholesterol agents for dual treatment of glioblastoma.
Assuntos
Alcinos/farmacologia , Anticolesterolemiantes/farmacologia , Antineoplásicos/farmacologia , Carbamatos/farmacologia , Cicloexanos/farmacologia , Ácidos Heptanoicos/farmacologia , Transferases Intramoleculares/antagonistas & inibidores , Pirróis/farmacologia , Alcinos/síntese química , Alcinos/química , Inibidores da Angiogênese/síntese química , Inibidores da Angiogênese/química , Inibidores da Angiogênese/farmacologia , Animais , Anticolesterolemiantes/síntese química , Anticolesterolemiantes/química , Antineoplásicos/síntese química , Antineoplásicos/química , Atorvastatina , Encéfalo/irrigação sanguínea , Carbamatos/síntese química , Carbamatos/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Cricetinae , Cicloexanos/síntese química , Cicloexanos/química , Ensaios de Seleção de Medicamentos Antitumorais , Sinergismo Farmacológico , Células Endoteliais/citologia , Células Endoteliais/efeitos dos fármacos , Humanos , Inibidores de Hidroximetilglutaril-CoA Redutases/farmacologia , Microssomos Hepáticos/enzimologia , Neovascularização Patológica/patologia , Relação Estrutura-AtividadeRESUMO
G protein-coupled receptors (GPCRs) share a common architecture consisting of seven transmembrane (TM) domains. Various lines of evidence suggest that this fold provides a generic binding pocket within the TM region for hosting agonists, antagonists, and allosteric modulators. Hence, an automated method was developed that allows a fast analysis and comparison of these generic ligand binding pockets across the entire GPCR family by providing the relevant information for all GPCRs in the same format. This methodology compiles amino acids lining the TM binding pocket including parts of the ECL2 loop in a so-called 1D ligand binding pocket vector and translates these 1D vectors in a second step into 3D receptor pharmacophore models. It aims to support various aspects of GPCR drug discovery in the pharmaceutical industry. Applications of pharmacophore similarity analysis of these 1D LPVs include definition of receptor subfamilies, prediction of species differences within subfamilies in regard to in vitro pharmacology and identification of nearest neighbors for GPCRs of interest to generate starting points for GPCR lead identification programs. These aspects of GPCR research are exemplified in the field of melanopsins, trace amine-associated receptors and somatostatin receptor subtype 5. In addition, it is demonstrated how 3D pharmacophore models of the LPVs can support the prediction of amino acids involved in ligand recognition, the understanding of mutational data in a 3D context and the elucidation of binding modes for GPCR ligands and their evaluation. Furthermore, guidance through 3D receptor pharmacophore modeling for the synthesis of subtype-specific GPCR ligands will be reported. Illustrative examples are taken from the GPCR family class C, metabotropic glutamate receptors 1 and 5 and sweet taste receptors, and from the GPCR class A, e.g. nicotinic acid and 5-hydroxytryptamine 5A receptor.
Assuntos
Descoberta de Drogas/métodos , Receptores Acoplados a Proteínas G/química , Sequência de Aminoácidos , Sítios de Ligação , Humanos , Ligantes , Modelos Moleculares , Dados de Sequência Molecular , Conformação Proteica , Receptores Acoplados a Proteínas G/antagonistas & inibidores , Receptores Acoplados a Proteínas G/metabolismoRESUMO
A series of branched and unbranched anilinohexafluoroisopropanols related to the known sulfonamide T0901317 were prepared and evaluated as activators/modulators of both LXRalpha and LXRbeta. A structure-activity relationship was established and compounds with high potency on both the receptors were identified. Many compounds showed a tendency toward selectivity for LXRbeta versus LXRalpha. Several analogues were evaluated for effects on plasma lipoprotein levels in mice. A few of these significantly raised HDL-cholesterol levels in plasma but showed markedly different effects on liver triglyceride content, suggesting that this series may yield candidates with improved efficacy/safety profiles compared to existing molecules.
Assuntos
Compostos de Anilina/síntese química , Proteínas de Ligação a DNA/efeitos dos fármacos , Receptores Citoplasmáticos e Nucleares/efeitos dos fármacos , Compostos de Anilina/farmacocinética , Compostos de Anilina/farmacologia , Animais , Aterosclerose/tratamento farmacológico , HDL-Colesterol/sangue , Lipoproteínas/sangue , Fígado , Receptores X do Fígado , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Receptores Nucleares Órfãos , Propanóis/síntese química , Propanóis/farmacocinética , Propanóis/farmacologia , Relação Estrutura-Atividade , Ativação Transcricional/efeitos dos fármacos , Triglicerídeos/sangueRESUMO
The solid-phase synthesis of substituted 1,2,4-triazoles tethered to a 4-mercaptopyrrolidine core 1 is described. This novel class of non-peptidic, Zn(2+) metallo-protease inhibitors was found to have inhibitory activity for the endothelin converting enzyme (ECE-1). The SAR of the substitution pattern in 1 is discussed.
Assuntos
Ácido Aspártico Endopeptidases/antagonistas & inibidores , Inibidores de Proteases/química , Inibidores de Proteases/síntese química , Pirrolidinas/química , Pirrolidinas/síntese química , Triazóis/química , Triazóis/síntese química , Enzimas Conversoras de Endotelina , Humanos , Concentração Inibidora 50 , Metaloendopeptidases/antagonistas & inibidores , Inibidores de Proteases/farmacologia , Pirrolidinas/farmacologia , Relação Estrutura-Atividade , Triazóis/farmacologiaRESUMO
We tested the hypothesis that endothelin-converting enzyme (ECE) inhibition ameliorates end-organ damage in rats harboring both human renin and human angiotensinogen genes (dTGR). Hypertension develops in the animals, and they die by age 7 weeks of heart and kidney failure. Three groups were studied: dTGR (n=12) receiving vehicle, dTGR receiving ECE inhibitor (RO0687629; 30 mg/kg by gavage; n=10), and Sprague-Dawley control rats (SD; n=10) receiving vehicle, all after week 4, with euthanasia at week 7. Systolic blood pressure was not reduced by ECE inhibitor compared with dTGR (205+/-6 versus 206+/-6 mm Hg at week 7, respectively). In contrast, ECE inhibitor treatment significantly reduced mortality rate to 20% (2 of 10), whereas untreated dTGR had a 52% mortality rate (7 of 12). ECE inhibitor treatment ameliorated cardiac damage and reduced left ventricular ECE activity below SD levels. Echocardiography at week 7 showed reduced cardiac hypertrophy (4.8+/-0.2 versus 5.7+/-0.2 mg/g, P<0.01) and increased left ventricular cavity diameter (5.5+/-0.3 versus 3.1+/-0.1 mm, P<0.001) and filling volume (0.42+/-0.04 versus 0.16+/-0.06 mL, P<0.05) after ECE inhibitor compared with untreated dTGR. ECE inhibitor treatment also reduced cardiac fibrosis, tissue factor expression, left ventricular basic fibroblast growth factor mRNA levels, and immunostaining in the vessel wall, independent of high blood pressure. In contrast, the ECE inhibitor treatment showed no renoprotective effect. These data are the first to show that ECE inhibition reduces angiotensin II-induced cardiac damage.