RESUMO
11ß-Hydroxysteroid dehydrogenase type 1 (11ß-HSD1) is implicated in the etiology of metabolic syndrome. We previously showed that pharmacological inhibition of 11ß-HSD1 ameliorated multiple facets of metabolic syndrome and attenuated atherosclerosis in ApoE-/- mice. However, the molecular mechanism underlying the atheroprotective effect was not clear. In this study, we tested whether and how 11ß-HSD1 inhibition affects vascular inflammation, a major culprit for atherosclerosis and its associated complications. ApoE-/- mice were treated with an 11ß-HSD1 inhibitor for various periods of time. Plasma lipids and aortic cholesterol accumulation were quantified. Several microarray studies were carried out to examine the effect of 11ß-HSD1 inhibition on gene expression in atherosclerotic tissues. Our data suggest 11ß-HSD1 inhibition can directly modulate atherosclerotic plaques and attenuate atherosclerosis independently of lipid lowering effects. We identified immune response genes as the category of mRNA most significantly suppressed by 11ß-HSD1 inhibition. This anti-inflammatory effect was further confirmed in plaque macrophages and smooth muscle cells procured by laser capture microdissection. These findings in the vascular wall were corroborated by reduction in circulating MCP1 levels after 11ß-HSD1 inhibition. Taken together, our data suggest 11ß-HSD1 inhibition regulates proinflammatory gene expression in atherosclerotic tissues of ApoE-/- mice, and this effect may contribute to the attenuation of atherosclerosis in these animals.
Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/antagonistas & inibidores , Aterosclerose/tratamento farmacológico , Inibidores Enzimáticos/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Vasculite/tratamento farmacológico , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/metabolismo , Animais , Apolipoproteínas E/genética , Aterosclerose/etiologia , Colesterol/metabolismo , Perfilação da Expressão Gênica , Genes MHC da Classe II/genética , Glucocorticoides/metabolismo , Microdissecção e Captura a Laser , Lipídeos/sangue , Camundongos , Camundongos Knockout , Análise em Microsséries , Vasculite/complicaçõesRESUMO
Metabolic syndrome is a combination of medical disorders that increases the risk of developing cardiovascular disease and diabetes. Constitutive overexpression of 11ß-HSD1 in adipose tissue in mice leads to metabolic syndrome. In the process of generating transgenic mice overexpressing 11ß-HSD1 in an inducible manner, we found a metabolic syndrome phenotype in control, transgenic mice, expressing the reverse tetracycline-transactivator (rtTA) in adipose tissue. The control mice exhibited all four sequelae of metabolic syndrome (visceral obesity, insulin resistance, dyslipidemia, and hypertension), a pro-inflammatory state and marked hepatic steatosis. Gene expression profiling of the adipose tissue, muscle and liver of these mice revealed changes in expression of genes involved in lipid metabolism, insulin resistance, and inflammation. Transient transfection of rtTA, but not tTS, into 3T3-L1 cells resulted in lipid accumulation. We conclude that expression of rtTA in adipose tissue causes metabolic syndrome in mice.
Assuntos
Tecido Adiposo/metabolismo , Síndrome Metabólica/genética , Transativadores/metabolismo , Ativação Transcricional , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/genética , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/metabolismo , Células 3T3-L1 , Tecido Adiposo/patologia , Animais , Pressão Sanguínea , Cromossomos de Mamíferos/genética , Cromossomos de Mamíferos/metabolismo , Fragmentação do DNA , Fígado Gorduroso/genética , Fígado Gorduroso/metabolismo , Fígado Gorduroso/patologia , Perfilação da Expressão Gênica , Resistência à Insulina , Metabolismo dos Lipídeos , Masculino , Síndrome Metabólica/metabolismo , Síndrome Metabólica/patologia , Camundongos , Camundongos Transgênicos , Músculos/metabolismo , Músculos/patologia , Fenótipo , Tetraciclina/metabolismo , Transativadores/genética , Transfecção , TransgenesRESUMO
Pre-exposure oral prophylaxis with antiviral drugs is a potential method for preventing transmission of human immunodeficiency virus type 1 (HIV-1). We show that oral delivery of CMPD167, a small molecule that binds to the CCR5 coreceptor, for 10-14 d can protect a substantial proportion of macaques from vaginal infection with a CCR5-using virus (SHIV-162P3). The macaques that became infected despite receiving CMPD167 had reduced plasma viremia levels during the earliest stages of infection.
Assuntos
Antagonistas dos Receptores CCR5 , Macaca/virologia , Pirazóis/farmacologia , Síndrome de Imunodeficiência Adquirida dos Símios/prevenção & controle , Vírus da Imunodeficiência Símia/efeitos dos fármacos , Vagina/virologia , Valina/análogos & derivados , Administração Oral , Animais , Feminino , Pirazóis/administração & dosagem , Fatores de Tempo , Vagina/efeitos dos fármacos , Valina/administração & dosagem , Valina/farmacologia , Carga ViralRESUMO
The enzyme 11beta-hydroxysteroid dehydrogenase (HSD) type 1 converts inactive cortisone into active cortisol in cells, thereby raising the effective glucocorticoid (GC) tone above serum levels. We report that pharmacologic inhibition of 11beta-HSD1 has a therapeutic effect in mouse models of metabolic syndrome. Administration of a selective, potent 11beta-HSD1 inhibitor lowered body weight, insulin, fasting glucose, triglycerides, and cholesterol in diet-induced obese mice and lowered fasting glucose, insulin, glucagon, triglycerides, and free fatty acids, as well as improved glucose tolerance, in a mouse model of type 2 diabetes. Most importantly, inhibition of 11beta-HSD1 slowed plaque progression in a murine model of atherosclerosis, the key clinical sequela of metabolic syndrome. Mice with a targeted deletion of apolipoprotein E exhibited 84% less accumulation of aortic total cholesterol, as well as lower serum cholesterol and triglycerides, when treated with an 11beta-HSD1 inhibitor. These data provide the first evidence that pharmacologic inhibition of intracellular GC activation can effectively treat atherosclerosis, the key clinical consequence of metabolic syndrome, in addition to its salutary effect on multiple aspects of the metabolic syndrome itself.
Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/antagonistas & inibidores , Adamantano/análogos & derivados , Arteriosclerose/tratamento farmacológico , Azepinas/administração & dosagem , Ativação Enzimática/efeitos dos fármacos , Inibidores Enzimáticos/administração & dosagem , Resistência à Insulina , Triazóis/administração & dosagem , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/metabolismo , Adamantano/administração & dosagem , Animais , Aorta/metabolismo , Arteriosclerose/complicações , Arteriosclerose/enzimologia , Glicemia/efeitos dos fármacos , Cortisona/metabolismo , Dieta Aterogênica , Modelos Animais de Doenças , Ácidos Graxos/sangue , Hidrocortisona , Insulina/sangue , Masculino , Camundongos , Camundongos Endogâmicos ICR , Camundongos Knockout , Síndrome , Triglicerídeos/sangueRESUMO
Human immunodeficiency virus type 1 (HIV-1) continues to spread, principally by heterosexual sex, but no vaccine is available. Hence, alternative prevention methods are needed to supplement educational and behavioural-modification programmes. One such approach is a vaginal microbicide: the application of inhibitory compounds before intercourse. Here, we have evaluated the microbicide concept using the rhesus macaque 'high dose' vaginal transmission model with a CCR5-receptor-using simian-human immunodeficiency virus (SHIV-162P3) and three compounds that inhibit different stages of the virus-cell attachment and entry process. These compounds are BMS-378806, a small molecule that binds the viral gp120 glycoprotein and prevents its attachment to the CD4 and CCR5 receptors, CMPD167, a small molecule that binds to CCR5 to inhibit gp120 association, and C52L, a bacterially expressed peptide inhibitor of gp41-mediated fusion. In vitro, all three compounds inhibit infection of T cells and cervical tissue explants, and C52L acts synergistically with CMPD167 or BMS-378806 to inhibit infection of cell lines. In vivo, significant protection was achieved using each compound alone and in combinations. CMPD167 and BMS-378806 were protective even when applied 6 h before challenge.
Assuntos
Fármacos Anti-HIV/farmacologia , Infecções por HIV/prevenção & controle , HIV/efeitos dos fármacos , Macaca mulatta/virologia , Fusão de Membrana/efeitos dos fármacos , Síndrome de Imunodeficiência Adquirida dos Símios/prevenção & controle , Vagina/virologia , Administração Intravaginal , Animais , Fármacos Anti-HIV/administração & dosagem , Antagonistas dos Receptores CCR5 , Antígenos CD4/metabolismo , Fusão Celular , Quimioterapia Combinada , Feminino , HIV/metabolismo , Proteína gp120 do Envelope de HIV/metabolismo , Infecções por HIV/transmissão , Infecções por HIV/virologia , Piperazinas/administração & dosagem , Piperazinas/farmacologia , Pirazóis/administração & dosagem , Pirazóis/farmacologia , Receptores CCR5/metabolismo , Receptores Virais/antagonistas & inibidores , Receptores Virais/metabolismo , Síndrome de Imunodeficiência Adquirida dos Símios/transmissão , Síndrome de Imunodeficiência Adquirida dos Símios/virologia , Vírus da Imunodeficiência Símia/efeitos dos fármacos , Vírus da Imunodeficiência Símia/metabolismo , Fatores de Tempo , Vagina/efeitos dos fármacos , Valina/administração & dosagem , Valina/análogos & derivados , Valina/farmacologiaRESUMO
Human immunodeficiency virus type 1 (HIV-1) fuses with cells after sequential interactions between its envelope glycoproteins, CD4 and a coreceptor, usually CC chemokine receptor 5 (CCR5) or CXC receptor 4 (CXCR4). CMPD 167 is a CCR5-specific small molecule with potent antiviral activity in vitro. We show that CMPD 167 caused a rapid and substantial (4-200-fold) decrease in plasma viremia in six rhesus macaques chronically infected with simian immunodeficiency virus (SIV) strains SIVmac251 or SIVB670, but not in an animal infected with the X4 simian-human immunodeficiency virus (SHIV), SHIV-89.6P. In three of the SIV-infected animals, viremia reduction was sustained. In one, there was a rapid, but partial, rebound and in another, there was a rapid and complete rebound. There was a substantial delay (>21 d) between the end of therapy and the onset of full viremia rebound in two animals. We also evaluated whether vaginal administration of gel-formulated CMPD 167 could prevent vaginal transmission of the R5 virus, SHIV-162P4. Complete protection occurred in only 2 of 11 animals, but early viral replication was significantly less in the 11 CMPD 167-recipients than in 9 controls receiving carrier gel. These findings support the development of small molecule CCR5 inhibitors as antiviral therapies, and possibly as components of a topical microbicide to prevent HIV-1 sexual transmission.
Assuntos
Antirretrovirais/farmacologia , Antagonistas dos Receptores CCR5 , Macaca mulatta/virologia , Pirazóis/farmacologia , Síndrome de Imunodeficiência Adquirida dos Símios/tratamento farmacológico , Vírus da Imunodeficiência Símia/efeitos dos fármacos , Valina/farmacologia , Animais , Linfócitos T CD4-Positivos/efeitos dos fármacos , Síndrome de Imunodeficiência Adquirida dos Símios/terapia , Síndrome de Imunodeficiência Adquirida dos Símios/transmissão , Valina/análogos & derivadosRESUMO
[reaction: see text] A novel approach to alpha,alpha-disubstituted-beta-amino acids (beta(2,2)-amino acids) was employed in the synthesis of a series of 3-(pyrrolidin-1-yl)propionic acids possessing high affinity for the CCR5 receptor and potent anti-HIV activity. The rat pharmacokinetics for these new analogues featured higher bioavailabilities and lower rates of clearance as compared to cyclopentane 1.
Assuntos
Fármacos Anti-HIV/farmacologia , Antagonistas dos Receptores CCR5 , Propionatos/farmacologia , Pirrolidinas/farmacologia , Fármacos Anti-HIV/farmacocinética , Disponibilidade Biológica , Propionatos/farmacocinética , Pirrolidinas/farmacocinéticaRESUMO
A series of racemic and homochiral alpha-aminothiazole-gamma-aminobutyroamides that display high affinities for human and murine CCR2 and functional antagonism by inhibition of monocyte recruitment are described. A representative example is (2S)-2-[2-(acetylamino)-1,3-thiazol-4-yl]-N-[3-methyl-5-(trifluoromethyl)benzyl]-4-(4-phenylpiperidin-1-yl)butanamide, which shows 5 nM affinity for human monocytes and CHO cells expressing the human CCR2b receptor. It also inhibited MCP-1 initiated chemotaxis of human monocytes with an IC50 of 0.69 nM.
Assuntos
Receptores de Quimiocinas/antagonistas & inibidores , Animais , Células CHO , Quimiocina CCL2/antagonistas & inibidores , Quimiocina CCL2/metabolismo , Quimiotaxia/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Cricetinae , Cricetulus , Cães , Desenho de Fármacos , Humanos , Técnicas In Vitro , Indicadores e Reagentes , Membranas/metabolismo , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , Antagonistas dos Receptores de Neurocinina-1 , Ratos , Ratos Sprague-Dawley , Receptores CCR2 , Receptores de Quimiocinas/efeitos dos fármacos , Estereoisomerismo , Relação Estrutura-AtividadeRESUMO
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
Synthesis of analogs containing more rigid bicyclic piperidine replacements for the 4-benzyloxycarbonyl-(ethyl)amino-piperidine moiety of the CCR5 antagonist structure, 1, is described. Although similar binding affinity to the lead was achieved with some analogs they were overall less potent anti-HIV agents suggesting that other features besides CCR5 binding are required for good anti-viral activity.
Assuntos
Fármacos Anti-HIV/síntese química , Antagonistas dos Receptores CCR5 , Sulfonas/síntese química , Fármacos Anti-HIV/farmacologia , Butanos/síntese química , Butanos/farmacologia , Relação Dose-Resposta a Droga , Concentração Inibidora 50 , Piperidinas/síntese química , Piperidinas/farmacologia , Relação Estrutura-Atividade , Sulfonas/farmacologia , Vírus/efeitos dos fármacosRESUMO
A series of 1,3,4-trisubstituted pyrrolidine CCR5 receptor antagonists containing a variety of fused heterocycles at the 4-position of the piperidine side chain has been discovered, which are orally bioavailable with potent anti-HIV activity.
Assuntos
Fármacos Anti-HIV/química , Antagonistas dos Receptores CCR5 , Compostos Heterocíclicos/química , Pirrolidinas/química , Administração Oral , Animais , Fármacos Anti-HIV/farmacocinética , Células HeLa , Compostos Heterocíclicos/farmacocinética , Humanos , Pirrolidinas/farmacocinética , Ratos , Receptores CCR5/metabolismoRESUMO
Human immunodeficiency virus type 1 (HIV-1) fusion with its target cells is initiated by sequential interactions between its envelope glycoprotein, CD4, and a co-receptor, usually CCR5 or CXCR4. Small molecules that bind to CCR5 and prevent its use by R5 HIV-1 strains are now being developed clinically as antiviral drugs. To test whether a block to CCR5 promotes the replication of viruses that enter cells via CXCR4 and are associated with accelerated disease progression, we administered a small molecule CCR5 inhibitor, CMPD 167, to three macaques dual-infected with both R5 (SIVmac251) and X4 (SHIV-89.6P) viruses. CMPD 167 caused a rapid and substantial (on average, 50-fold) suppression of R5 virus replication in each animal. In two of the animals, but not in the third, a rapid, transient, 8- to 15-fold increase in the amount of plasma X4 virus occurred. In neither animal was the increase in X4 viral load sustained throughout therapy, however. These observations may have relevance for the development of CCR5 inhibitors for treatment of HIV-1 infection of humans.
Assuntos
Fármacos Anti-HIV/uso terapêutico , Antagonistas dos Receptores CCR5 , Infecções por HIV/tratamento farmacológico , HIV-1 , Pirazóis/uso terapêutico , Vírus Reordenados , Síndrome de Imunodeficiência Adquirida dos Símios/tratamento farmacológico , Vírus da Imunodeficiência Símia , Valina/uso terapêutico , Animais , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Infecções por HIV/virologia , HIV-1/fisiologia , Macaca mulatta , Vírus Reordenados/fisiologia , Síndrome de Imunodeficiência Adquirida dos Símios/virologia , Vírus da Imunodeficiência Símia/genética , Vírus da Imunodeficiência Símia/patogenicidade , Valina/análogos & derivados , Carga Viral , Replicação Viral/efeitos dos fármacosRESUMO
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
Replacement of the flexible connecting chains between the piperidine moiety and an aromatic group in previous CCR5 antagonists with heterocycles, such as pyrazole and isoxazole, provided potent CCR5 antagonists with excellent anti-HIV-1 activity in vitro. SAR studies revealed optimal placement of an unsubstituted nitrogen atom in the heterocycle to be meta to the bond connected to the 4-position of piperidine. Truncation of a benzyl group to a phenyl group afforded compounds with dramatically improved oral bioavailability, albeit with reduced activity.
Assuntos
Fármacos Anti-HIV/síntese química , Fármacos Anti-HIV/farmacocinética , Antagonistas dos Receptores CCR5 , Piperidinas/síntese química , Piperidinas/farmacocinética , Animais , Fármacos Anti-HIV/química , Divisão Celular/efeitos dos fármacos , Células HeLa , Humanos , Estrutura Molecular , Piperidinas/química , Pirazóis/química , Pirazóis/farmacocinética , Ratos , Relação Estrutura-AtividadeRESUMO
Efforts toward the exploration of the title compounds as CCR5 antagonists are disclosed. The basis for such work stems from the fact that cellular proliferation of HIV-1 requires the cooperative assistance of both CCR5 and CD4 receptors. The synthesis and SAR of pyrrolidineacetic acid derivatives as CCR5 antagonists displaying potent binding and antiviral properties in a HeLa cell-based HIV-1 infectivity assay are discussed.
Assuntos
Fármacos Anti-HIV/síntese química , Antagonistas dos Receptores CCR5 , HIV-1/efeitos dos fármacos , Pirrolidinas/síntese química , Acetatos/química , Fármacos Anti-HIV/farmacologia , Sítios de Ligação , Divisão Celular/efeitos dos fármacos , Células HeLa , Humanos , Piperidinas/síntese química , Piperidinas/farmacologia , Pirrolidinas/química , Relação Estrutura-AtividadeRESUMO
A series of CCR5 antagonists containing bicyclic isoxazolidines was generated through a nitrone mediated cycloaddition with olefins bearing the preferred pharmacophores previously described. Potent antagonists (3 and 16) were generated with enhanced affinity for the CCR5 receptor while maintaining antiviral activity against HIV.
Assuntos
Fármacos Anti-HIV/síntese química , Antagonistas dos Receptores CCR5 , Isoxazóis/síntese química , Fármacos Anti-HIV/farmacologia , Compostos Bicíclicos Heterocíclicos com Pontes/síntese química , Compostos Bicíclicos Heterocíclicos com Pontes/química , Compostos Bicíclicos Heterocíclicos com Pontes/farmacologia , HIV-1/efeitos dos fármacos , Humanos , Isoxazóis/química , Isoxazóis/farmacologia , Testes de Sensibilidade Microbiana , Conformação Molecular , Ligação Proteica , Pirrolidinas/síntese química , Pirrolidinas/química , Pirrolidinas/farmacologia , Relação Estrutura-AtividadeRESUMO
Incorporation of acidic functional groups into a lead CCR5 antagonist identified from a targeted combinatorial library resulted in compounds with enhanced anti-HIV-1 activity and attenuated L-type calcium channel affinity.
Assuntos
Fármacos Anti-HIV/síntese química , Fármacos Anti-HIV/farmacologia , Antagonistas dos Receptores CCR5 , HIV-1/efeitos dos fármacos , Animais , Fármacos Anti-HIV/farmacocinética , Células CHO , Canais de Cálcio Tipo L/efeitos dos fármacos , Fenômenos Químicos , Físico-Química , Quimiocina CCL4 , Cricetinae , Meia-Vida , Células HeLa , Humanos , Proteínas Inflamatórias de Macrófagos/antagonistas & inibidores , Ratos , Receptores CCR5/química , Relação Estrutura-AtividadeRESUMO
The results of investigations in these laboratories of 2-aryl-4-(piperidin-1-yl)butanamines and 1,3,4-trisubstituted pyrrolidines as human CCR5 antagonists have recently been disclosed. To facilitate further development of these antagonists, we have developed a pharmacophore model based on the structure-activity relationships (SAR) and a human CCR5 receptor docking model using the crystal structure of rhodopsin as a template [Palczewski, K., et al. (2000) Science 289, 739-745]. Guided by the receptor docking model, we have mapped the compounds' site of interaction with CCR5 using site-directed mutagenesis experiments. Our results are consistent with a binding site for the two series that is located within a cavity near the extracellular surface formed by transmembrane helices 2, 3, 6, and 7. This site is overlapping yet distinct from that reported for another antiviral agent which binds to CCR5 [Dragic, T., et al. (2000) Proc. Natl. Acad. Sci. U.S.A. 97, 5639-5644].
Assuntos
Butanos/química , Antagonistas dos Receptores CCR5 , Modelos Moleculares , Mutagênese Sítio-Dirigida , Piperidinas/química , Pirrolidinas/química , Receptores CCR5/química , Alanina/genética , Amidas/química , Sequência de Aminoácidos , Substituição de Aminoácidos/genética , Animais , Sítios de Ligação/genética , Ligação Competitiva/genética , Células CHO , Bovinos , Cricetinae , Humanos , Dados de Sequência Molecular , Estrutura Secundária de Proteína/genética , Compostos de Amônio Quaternário/química , Receptores CCR5/biossíntese , Receptores CCR5/genética , Rodopsina/química , Homologia de Sequência de Aminoácidos , Relação Estrutura-AtividadeRESUMO
Modifications of the alkyl acetic acid portion and the phenyl on pyrrolidine in our lead pyrazole compound 1 afforded the isopropyl compound 9. This compound is a potent CCR5 antagonist showing good in vitro antiviral activity against HIV-1, an excellent selectivity profile, and good oral bioavailability in three animal species. During this investigation, a new method for the preparation of alpha-(pyrrolidin-1-yl)-alpha,alpha-dialkyl acetic acid from a pyrrolidine and alpha-bromo-alpha,alpha-dialkyl acetic acid using silver triflate was discovered. This allowed us to prepare compounds such as 24 and 25 for the first time. A novel Pd-mediated N-dealkylation of alpha-(pyrrolidin-1-yl)acetic acid was also uncovered.
Assuntos
Fármacos Anti-HIV/síntese química , Fármacos Anti-HIV/farmacocinética , Antagonistas dos Receptores CCR5 , Piperidinas/síntese química , Piperidinas/farmacocinética , Acetatos/química , Acetatos/farmacocinética , Administração Oral , Animais , Fármacos Anti-HIV/química , Disponibilidade Biológica , Cães , Células HeLa , Humanos , Macaca mulatta , Estrutura Molecular , Monócitos/efeitos dos fármacos , Piperidinas/química , Pirazóis/química , Pirazóis/farmacocinética , Ratos , Relação Estrutura-AtividadeRESUMO
Extensive SAR studies in our benzylpyrazole series of CCR5 antagonists have shown that both lipophilic and hydrophilic substituents on the phenyl of the benzyl group increase antiviral potency. However, improvements in pharmacokinetic profiles were generally only observed with more lipophilic substitutions. 4-Biphenyl (51) performed the best in this regard. Highly lipophilic substituents impart undesirable ion channel activity to these CCR5 antagonists. Alkoxy substituents provide a good balance of antiviral activity, pharmacokinetic parameters, and selectivity. Compounds 42b and 42d, containing a 3,4-dimethoxy substituent, are considered the most promising improvements over parent compounds 9. They demonstrate improved antiviral activity while retaining good pharmacokinetic profile and selectivity.