RESUMO
The discovery and optimization of potent and selective aminobenzimidazole glucagon receptor antagonists are described. One compound possessing moderate pharmacokinetic properties in multiple preclinical species was orally efficacious at inhibiting glucagon-mediated glucose excursion in transgenic mice expressing the human glucagon receptor, and in rhesus monkeys. The compound also significantly lowered glucose levels in a murine model of diabetes.
Assuntos
Benzimidazóis/química , Receptores de Glucagon/antagonistas & inibidores , Receptores de Glucagon/química , Administração Oral , Animais , Benzimidazóis/farmacocinética , Células CHO , Química Farmacêutica/métodos , Cricetinae , Cricetulus , Diabetes Mellitus Experimental/metabolismo , Glucagon/química , Humanos , Concentração Inibidora 50 , Macaca mulatta , Camundongos , Camundongos TransgênicosRESUMO
Pharmacologic antagonism of CCR5, a chemokine receptor expressed on macrophages and activated T cells, is an effective antiviral therapy in patients with macrophage-tropic HIV infection, but its efficacy in modulating inflammation and immunity is only just beginning to be investigated. In this regard, the recruitment of CCR5-bearing cells into clinical allografts is a hallmark of acute rejection and may anticipate chronic rejection, whereas conventionally immunosuppressed renal transplant patients homozygous for a nonfunctional Delta32 CCR5 receptor rarely exhibit late graft loss. Therefore, we explored the effects of a potent, highly selective CCR5 antagonist, Merck's compound 167 (CMPD 167), in an established cynomolgus monkey cardiac allograft model. Although perioperative stress responses (fever, diminished activity) and the recruitment of CCR5-bearing leukocytes into the graft were markedly attenuated, anti-CCR5 monotherapy only marginally prolonged allograft survival. In contrast, relative to cyclosporine A monotherapy, CMPD 167 with cyclosporine A delayed alloantibody production, suppressed cardiac allograft vasculopathy, and tended to further prolong graft survival. CCR5 therefore represents an attractive therapeutic target for attenuating postsurgical stress responses and favorably modulating pathogenic alloimmunity in primates, including man.
Assuntos
Antagonistas dos Receptores CCR5 , Sobrevivência de Enxerto/efeitos dos fármacos , Transplante de Coração/imunologia , Macrófagos/imunologia , Pirazóis/administração & dosagem , Linfócitos T/imunologia , Tolerância ao Transplante/efeitos dos fármacos , Valina/análogos & derivados , Animais , Formação de Anticorpos/efeitos dos fármacos , Formação de Anticorpos/imunologia , Autoimunidade/efeitos dos fármacos , Autoimunidade/imunologia , Ciclosporina/administração & dosagem , Modelos Animais de Doenças , Sobrevivência de Enxerto/imunologia , Infecções por HIV/tratamento farmacológico , Infecções por HIV/imunologia , Infecções por HIV/patologia , Transplante de Coração/patologia , Humanos , Imunossupressores/administração & dosagem , Inflamação/tratamento farmacológico , Inflamação/imunologia , Inflamação/patologia , Isoanticorpos/imunologia , Transplante de Rim/imunologia , Macaca fascicularis , Macrófagos/patologia , Masculino , Estresse Fisiológico/tratamento farmacológico , Estresse Fisiológico/imunologia , Estresse Fisiológico/patologia , Linfócitos T/patologia , Tolerância ao Transplante/imunologia , Transplante Homólogo , Valina/administração & dosagem , Doenças Vasculares/tratamento farmacológico , Doenças Vasculares/imunologia , Doenças Vasculares/patologiaRESUMO
MK-0767, (+/-)-5-[(2,4-dioxothiazolidin-5-yl)methyl]-2-methoxy-N-[[(4-trifluoromethyl)phenyl]methyl]benzamide, is a thiazolidinedione-containing dual peroxisome proliferator-activated receptor (PPAR) alpha/gamma agonist that has been studied as a potential treatment for patients with type 2 diabetes. MK-0767 contains a chiral center at the C-5 position of the thiazolidinedione ring and was being developed as the racemate, due to the rapid interconversion of its enantiomers in biological samples. In the present work the in vitro and in vivo concentration ratios of the (+)-(R) to (-)-(S) enantiomers of MK-0767 were determined in plasma from humans (in vitro only) and nonclinical species used in the toxicological evaluation of rac-MK-0767, namely CD-1 mice, Sprague-Dawley rats, beagle dogs, New Zealand white rabbits, and rhesus monkeys. The R/S ratio was determined by chiral liquid chromatography/tandem mass spectrometry. Species differences were observed in the in vitro and in vivo enantiomeric ratios, as well as differences between in vitro and in vivo in some species. The in vitro R/S ratio was similar in dogs and humans (approximately 1.5-1.7). In rats and monkeys, the ratio was approximately unity, both in vitro and in vivo. In mice, the ratio was higher in vitro (approximately 1) than in vivo (approximately 0.6), while in rabbits it was higher in vivo (approximately 1) than in vitro (approximately 0.5). These results suggested that differential binding of the MK-0767 enantiomers to plasma and tissue proteins and other macromolecules may be affecting the R/S ratio both in vitro and in vivo, since in protein-free systems MK-0767 exists as the racemate.
Assuntos
Tiazóis/sangue , Tiazóis/química , Animais , Cães , Haplorrinos/sangue , Humanos , Camundongos , Estrutura Molecular , Coelhos , Ratos , Especificidade da Espécie , EstereoisomerismoRESUMO
The mechanisms of pharmacokinetic interactions of a novel anti-human immunodeficiency virus (anti-HIV-1) antagonist of chemokine receptor 5 (CCR5) [2-(R)-[N-methyl-N-(1-(R)-3-(S)-((4-(3-benzyl-1-ethyl-(1H)-pyrazol-5-yl)piperidin-1-yl)methyl)-4-(S)-(3-fluorophenyl)cyclopent-1-yl)amino]-3-methylbutanoic acid (MRK-1)] with ritonavir were evaluated in rats and monkeys. MRK-1 was a good substrate for the human (MDR1) and mouse (Mdr1a) multidrug resistance protein transporters and was metabolized by CYP3A isozymes in rat, monkey, and human liver microsomes. Both the in vitro MDR1-mediated transport and oxidative metabolism of MRK-1 were inhibited by ritonavir. Although the systemic pharmacokinetics of MRK-1 in rats and monkeys were linear, the oral bioavailability increased with an increase in dose from 2 to 10 mg/kg. The area under the plasma concentration-time curve (AUC) of MRK-1 was increased 4- to 6-fold when a 2 or 10 mg/kg dose was orally coadministered with 10 mg/kg ritonavir. Further pharmacokinetic studies in rats indicated that P-glycoprotein (P-gp) inhibition by ritonavir increased the intestinal absorption of 2 mg/kg MRK-1 maximally by approximately 30 to 40%, and a major component of the interaction likely resulted from its reduced systemic clearance via the inhibition of CYP3A isozymes. Oral coadministration of quinidine (10 and 30 mg/kg) increased both the extent and the first-order rate of absorption of MRK-1 (2 mg/kg) by approximately 40 to 50% and approximately 100 to 300%, respectively, in rats, thus further substantiating the role of P-gp in modulating the intestinal absorption of MRK-1 in this species. At the 10 mg/kg MRK-1 dose, however, the entire increase in its AUC upon coadministration with ritonavir or quinidine could be attributed to a reduced systemic clearance, and no effects on intestinal absorption were apparent. In contrast to rats, the effects of P-gp in determining the intestinal absorption of MRK-1 appeared less significant in rhesus monkeys at either dose.
Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/fisiologia , Hidrocarboneto de Aril Hidroxilases/fisiologia , Antagonistas dos Receptores CCR5 , Oxirredutases N-Desmetilantes/fisiologia , Pirazóis/metabolismo , Ritonavir/farmacologia , Valina/metabolismo , Administração Oral , Animais , Citocromo P-450 CYP3A , Interações Medicamentosas , Inibidores da Protease de HIV/farmacologia , Haplorrinos , Absorção Intestinal/efeitos dos fármacos , Masculino , Oxirredução , Ligação Proteica , Pirazóis/farmacocinética , Quinidina/farmacologia , Ratos , Ratos Sprague-Dawley , Valina/análogos & derivados , Valina/farmacocinéticaRESUMO
Eosinophils are major effector cells implicated in a number of chronic inflammatory diseases in humans, particularly bronchial asthma and allergic rhinitis. The beta-chemokine receptor C-C chemokine receptor 3 (CCR3) provides a mechanism for the selective recruitment of eosinophils into tissue and thus has recently become an attractive biological target for therapeutic intervention. In order to develop in vivo models of inflammatory diseases, it is essential to identify and characterize the homologues of human eotaxin (C-C chemokine ligand 11) and CCR3 from other species, such as non-human primates. Accordingly, we cloned the macaque eotaxin and CCR3 genes and revealed that they were 91 and 92% identical at the amino acid level to their human homologues, respectively. Macaque CCR3 expressed in the murine pre-B L1-2 cell line bound macaque eotaxin with high affinity (K(d) = 0.1 nm) and exhibited a robust eotaxin-induced Ca(2+) flux and chemotaxis. Characterization of beta-chemokines on native macaque CCR3 on eosinophils was performed by means of eotaxin-induced shape change in whole blood using a novel signaling assay known as gated autofluorescence forward scatter. Additionally, mAbs were raised against macaque CCR3 using two different immunogens: a 30-amino acid synthetic peptide derived from the predicted NH(2) terminus of macaque CCR3 and intact macaque CCR3-transfected cells. These anti-macaque CCR3 monoclonal antibodies exhibited potent antagonist activity in receptor binding and functional assays. The characterization of the macaque eotaxin/CCR3 axis and development of antagonistic anti-macaque CCR3 monoclonal antibodies will facilitate the development of CCR3 small molecule antagonists with the hope of ameliorating chronic inflammatory diseases in humans.
Assuntos
Anticorpos Monoclonais/biossíntese , Receptores de Quimiocinas/antagonistas & inibidores , Receptores de Quimiocinas/genética , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/uso terapêutico , Ligação Competitiva , Cálcio/metabolismo , Linhagem Celular , Quimiocina CCL11 , Quimiocina CCL5/fisiologia , Quimiocinas CC/genética , Quimiocinas CC/metabolismo , Quimiotaxia , Clonagem Molecular , Eosinófilos/fisiologia , Humanos , Macaca mulatta , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Receptores CCR3 , Receptores de Quimiocinas/químicaRESUMO
The pharmacokinetics and oral bioavailability of (R)-N-[4-[2-[[2-hydroxy-2-(pyridin-3-yl)ethyl]amino]ethyl]phenyl]-4-[4-[4-(trifluoromethylphenyl]thiazol-2-yl]benzenesulfonamide (1), a 3-pyridyl thiazole benzenesulfonamide beta3-adrenergic receptor agonist, were investigated in rats, dogs, and monkeys. Systemic clearance was higher in rats (approximately 30 ml/min/kg) than in dogs and monkeys (both approximately 10 ml/min/kg), and oral bioavailability was 17, 27, and 4%, respectively. Since systemic clearance was 25 to 40% of hepatic blood flow in these species, hepatic extraction was expected to be low, and it was likely that oral bioavailability was limited either by absorption or a large first-pass effect in the gut. The absorption and excretion of 3H-labeled 1 were investigated in rats, and only 28% of the administered radioactivity was orally absorbed. Subsequently, the hepatic extraction of 1 was evaluated in rats (30%) and monkeys (47%). The low oral bioavailability in rats could be explained completely by poor oral absorption and hepatic first-pass metabolism; in monkeys, oral absorption was either less than in rats or first-pass extraction in the gut was greater. In an attempt to increase oral exposure, the pharmacokinetics and oral bioavailability of two potential prodrugs of 1, an N-ethyl [(R)-N-[4-[2-[ethyl[2-hydroxy-2-(3-pyridinyl)ethyl]amino]ethyl]phenyl]-4-[4-[4-(trifluoromethyl)phenyl]thiazol-2-yl]benzenesulfonamide; 2] and a morpholine derivative [(R)-N-[4-[2-[2-(3-pyridinyl)morpholin-4-yl]ethyl]phenyl]-4-[4-[4-(trifluoromethyl)- phenyl]thiazol-2-yl]benzenesulfonamide; 3], were evaluated in monkeys. Conversion to 1 was low (<3%) with both derivatives, and neither entity was an effective prodrug, but the oral bioavailability of 3 (56%) compared with 1 (4%) was significantly improved. The hypothesis that the increased oral bioavailability of 3 was due to a reduction in hydrogen bonding sites in the molecule led to the design of (R)-N-[4-[2-[[2-hydroxy-2-(pyridin-2-yl)ethyl]amino]ethyl]phenyl]-4-[4-(4-trifluoromethylphenyl)thiazol-2-yl]benzenesulfonamide (4), a 2-pyridyl beta3-adrenergic receptor agonist with improved oral bioavailability in rats and monkeys.
Assuntos
Agonistas de Receptores Adrenérgicos beta 3 , Agonistas Adrenérgicos beta/farmacocinética , Sulfonamidas/farmacocinética , Tiazóis/farmacocinética , Administração Oral , Agonistas Adrenérgicos beta/química , Agonistas Adrenérgicos beta/metabolismo , Animais , Disponibilidade Biológica , Cães , Avaliação Pré-Clínica de Medicamentos , Macaca mulatta , Masculino , Ratos , Ratos Sprague-Dawley , Receptores Adrenérgicos beta 3/fisiologia , Sulfonamidas/química , Sulfonamidas/metabolismo , Tiazóis/química , Tiazóis/metabolismo , BenzenossulfonamidasRESUMO
Relationships inevitably develop between humans and animals, regardless of the function or use of the animal partners. The need to recognize the existence of these human-animal bonds, as well as acknowledge the use of the animals, is widespread. Religious memorial services for animals in certain areas of the world provide an historical basis for such acknowledgment activities. The diversity of sacred and secular approaches to memorializing or acknowledging animals is illustrated by representative examples of such events. The need to establish such events, particularly in academic and research settings, is emphasized. The pros and cons of developing and establishing acknowledgment activities in addition to the benefits of implementing such events are discussed.