RESUMO
Studies in lipoprotein kinetics almost exclusively rely on steady-state approaches to modeling. Herein, we have used a non-steady-state experimental design to examine the role of cholesteryl ester transfer protein (CETP) in mediating HDL-TG flux in vivo in rhesus macaques, and therefore, we developed an alternative strategy to model the data. Two isotopomers ([(2)H11] and [(13)C18]) of oleic acid were administered (orally and intravenously, respectively) to serve as precursors for labeling TGs in apoB-containing lipoproteins. The flux of a specific TG (52:2) from these donor lipoproteins to HDL was used as the measure of CETP activity; calculations are also presented to estimate total HDL-TG flux. Based on our data, we estimate that the peak total postprandial TG flux to HDL via CETP is â¼ 13 mg · h(-1) · kg(-1) and show that this transfer was inhibited by 97% following anacetrapib treatment. Collectively, these data demonstrate that HDL TG flux can be used as a measure of CETP activity in vivo. The fact that the donor lipoproteins can be labeled in situ using well-established stable isotope tracer techniques suggests ways to measure this activity for native lipoproteins in free-living subjects under any physiological conditions.
Assuntos
Proteínas de Transferência de Ésteres de Colesterol/antagonistas & inibidores , Proteínas de Transferência de Ésteres de Colesterol/metabolismo , Lipoproteínas HDL/metabolismo , Oxazolidinonas/farmacologia , Triglicerídeos/metabolismo , Animais , Lipoproteínas HDL/sangue , Macaca mulatta , Masculino , Modelos Biológicos , Triglicerídeos/sangueRESUMO
SAR studies of the substitution effect on the central phenyl ring of the biphenyl scaffold were carried out using anacetrapib (9a) as the benchmark. The results revealed that the new analogs with substitutions to replace trifluoromethyl (9a) had a significant impact on CETP inhibition in vitro. In fact, analogs with some small groups were as potent or more potent than the CF(3) derivative for CETP inhibition. Five of these new analogs raised HDL-C significantly (>20mg/dL). None of them however was better than anacetrapib in vivo. The synthesis and biological evaluation of these CETP inhibitors are described.
Assuntos
Proteínas de Transferência de Ésteres de Colesterol/antagonistas & inibidores , Oxazolidinonas/farmacologia , Animais , Química Farmacêutica/métodos , HDL-Colesterol/metabolismo , Relação Dose-Resposta a Droga , Desenho de Fármacos , Humanos , Concentração Inibidora 50 , Camundongos , Camundongos Transgênicos , Modelos Químicos , Relação Estrutura-AtividadeRESUMO
We describe structure-activity studies leading to the discovery of 2-arylbenzoxazole 3, the first in a series to raise serum high-density lipoprotein cholesterol levels in transgenic mice. Replacement of the 4-piperidinyloxy moiety with piperazinyl provided a more synthetically tractable lead, which upon optimization resulted in compound 4, an excellent inhibitor of cholesteryl ester transfer protein function with good pharmacokinetic properties and in vivo efficacy.
Assuntos
Acetanilidas/química , Benzoxazóis/química , Proteínas de Transferência de Ésteres de Colesterol/antagonistas & inibidores , HDL-Colesterol/sangue , Acetanilidas/síntese química , Acetanilidas/farmacocinética , Animais , Benzoxazóis/síntese química , Benzoxazóis/farmacocinética , Proteínas de Transferência de Ésteres de Colesterol/metabolismo , Camundongos , Camundongos Transgênicos , Relação Estrutura-AtividadeRESUMO
The development of a series of 2-arylbenzoxazole alpha-alkoxyamide and beta-alkoxyamine inhibitors of cholesteryl ester transfer protein (CETP) is described. Highly fluorinated alpha-alkoxyamides proved to be potent inhibitors of CETP in vitro, and the highly fluorinated 2-arylbenzoxazole beta-alkoxyamine 4 showed a desirable combination of in vitro potency (IC(50)=151 nM) and oral bioavailability in the mouse.
Assuntos
Benzoxazóis/síntese química , Benzoxazóis/metabolismo , Proteínas de Transferência de Ésteres de Colesterol/antagonistas & inibidores , Proteínas de Transferência de Ésteres de Colesterol/metabolismo , Álcoois/síntese química , Álcoois/metabolismo , Amidas/síntese química , Amidas/metabolismoRESUMO
A new class of CETP inhibitors was designed and prepared. These compounds are potent both in vitro and in vivo. The most active compound (12d) has shown an ability to raise HDL significantly in transgenic mouse PD model.
Assuntos
Benzilaminas/química , Compostos de Bifenilo/química , Carbamatos/química , Proteínas de Transferência de Ésteres de Colesterol/antagonistas & inibidores , Animais , Benzilaminas/síntese química , Benzilaminas/farmacologia , Compostos de Bifenilo/síntese química , Compostos de Bifenilo/farmacologia , Carbamatos/síntese química , Carbamatos/farmacologia , Proteínas de Transferência de Ésteres de Colesterol/metabolismo , Modelos Animais de Doenças , Desenho de Fármacos , Lipoproteínas HDL/metabolismo , Camundongos , Camundongos Transgênicos , Relação Estrutura-AtividadeRESUMO
One approach to understanding how orally administered drugs are absorbed and metabolized involves measuring compound concentrations in portal vein blood and in systemic circulation at various time points. In mice, blood samples are generally collected through terminal bleeding, a process that requires a large number of mice and is susceptible to variation between individuals. The authors developed a portal vein cannulation procedure for serial bleeding in the mouse, using a modified catheter containing a stainless steel stylet that is implanted directly in the portal vein. To demonstrate the technique, they orally administered two different compounds to mice and obtained blood samples from the tail vein and portal vein at different time points. They analyzed compound concentrations using liquid chromatography-tandem mass spectrometry. The technique refines existing methods for pharmacokinetic studies in the mouse and reduces the number of mice required.
Assuntos
Cateterismo Periférico/veterinária , Avaliação Pré-Clínica de Medicamentos/métodos , Veia Porta/cirurgia , Xenobióticos/administração & dosagem , Administração Oral , Animais , Peso Corporal/efeitos dos fármacos , Cateterismo Periférico/métodos , Cateteres de Demora/veterinária , Injeções Intravenosas , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Recuperação de Função Fisiológica/efeitos dos fármacos , Xenobióticos/farmacocinéticaRESUMO
The discovery of novel acyclic amide cannabinoid-1 receptor inverse agonists is described. They are potent, selective, orally bioavailable, and active in rodent models of food intake and body weight reduction. A major focus of the optimization process was to increase in vivo efficacy and to reduce the potential for formation of reactive metabolites. These efforts led to the identification of compound 48 for development as a clinical candidate for the treatment of obesity.
Assuntos
Fármacos Antiobesidade/farmacologia , Canabinoides/farmacologia , Obesidade/tratamento farmacológico , Receptor CB1 de Canabinoide/agonistas , Receptor CB2 de Canabinoide/agonistas , Animais , Fármacos Antiobesidade/síntese química , Fármacos Antiobesidade/química , Peso Corporal/efeitos dos fármacos , Canabinoides/síntese química , Canabinoides/química , AMP Cíclico/metabolismo , Ingestão de Alimentos/efeitos dos fármacos , Humanos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Microssomos/efeitos dos fármacos , Microssomos/metabolismo , Ratos , Receptor CB1 de Canabinoide/antagonistas & inibidores , Receptor CB1 de Canabinoide/metabolismo , Receptor CB2 de Canabinoide/antagonistas & inibidores , Receptor CB2 de Canabinoide/metabolismoRESUMO
The development of the structure-activity studies leading to the discovery of anacetrapib is described. These studies focused on varying the substitution of the oxazolidinone ring of the 5-aryloxazolidinone system. Specifically, it was found that substitution of the 4-position with a methyl group with the cis-stereochemistry relative to the 5-aryl group afforded compounds with increased cholesteryl ester transfer protein (CETP) inhibition potency and a robust in vivo effect on increasing HDL-C levels in transgenic mice expressing cynomolgus monkey CETP.
Assuntos
Proteínas de Transferência de Ésteres de Colesterol/antagonistas & inibidores , Oxazolidinonas/síntese química , Animais , Proteínas de Transferência de Ésteres de Colesterol/química , HDL-Colesterol/sangue , Humanos , Macaca fascicularis , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Oxazolidinonas/farmacocinética , Oxazolidinonas/farmacologia , Proteínas Recombinantes/química , Estereoisomerismo , Relação Estrutura-AtividadeRESUMO
The cannabinoid-1 receptor (CB1R) has been implicated in the control of energy balance. To explore the pharmacological utility of CB1R inhibition for the treatment of obesity, we evaluated the efficacy of N-[(1S,2S)-3-(4-chlorophenyl)-2-(3-cyanophenyl)-1-methylpropyl]-2-methyl-2-[[5-(trifluoromethyl)pyridin-2-yl]oxy]propanamide (MK-0364) and determined the relationship between efficacy and brain CB1R occupancy in rodents. MK-0364 was shown to be a highly potent CB1R inverse agonist that inhibited the binding and functional activity of various agonists with a binding K(i) of 0.13 nM for the human CB1R in vitro. MK-0364 dose-dependently inhibited food intake and weight gain, with an acute minimum effective dose of 1 mg/kg in diet-induced obese (DIO) rats. CB1R mechanism-based effect was demonstrated for MK-0364 by its lack of efficacy in CB1R-deficient mice. Chronic treatment of DIO rats with MK-0364 dose-dependently led to significant weight loss with a minimum effective dose of 0.3 mg/kg (p.o.), or a plasma C(max) of 87 nM. Weight loss was accompanied by the loss of fat mass. Partial occupancy (30-40%) of brain CB1R by MK-0364 was sufficient to reduce body weight. The magnitude of weight loss was correlated with brain CB1R occupancy. The partial receptor occupancy requirement for efficacy was also consistent with the reduced food intake of the heterozygous mice carrying one disrupted allele of CB1R gene compared with the wild-type mice. These studies demonstrated that MK-0364 is a highly potent and selective CB1R inverse agonist and that it is orally active in rodent models of obesity.