RESUMO
The SAR of 1-sulfonyl-cyclopentyl carboxylic acid amides, ligands for the VLA-4 integrin, was investigated. This effort resulted in the identification of N-(3-phenylsulfonyl-3-piperidinoyl)-(L)-4-(2',6'-dimethoxyphenyl)phenylalanine 52 as a potent, selective VLA-4 antagonist (IC(50)=90 pM). Expansion of the SAR demonstrated that this structural unit can be used to identify a diverse series of sub-nanomolar antagonists.
Assuntos
Integrina alfa4beta1/antagonistas & inibidores , Fenilalanina/análogos & derivados , Fenilalanina/farmacologia , Amidas/química , Amidas/metabolismo , Amidas/farmacologia , Moléculas de Adesão Celular , Humanos , Imunoglobulinas , Concentração Inibidora 50 , Integrina alfa4beta1/metabolismo , Células Jurkat , Mucoproteínas/antagonistas & inibidores , Fenilalanina/farmacocinética , Ensaio Radioligante , Receptores de Retorno de Linfócitos/antagonistas & inibidores , Relação Estrutura-Atividade , Molécula 1 de Adesão de Célula Vascular/metabolismoRESUMO
The design, synthesis, and biological evaluation of N-arylprolyl-dipeptide derivatives as small molecule VLA-4 antagonists is described. Potency against VLA-4 and alpha(4)beta(7) and rat pharmacokinetic evaluation revealed some advantages over the related N-(arylsulfonyl)-prolyl-dipeptide analogues.
Assuntos
Dipeptídeos/síntese química , Dipeptídeos/farmacologia , Integrina alfa4beta1/antagonistas & inibidores , Animais , Dipeptídeos/sangue , Dipeptídeos/farmacocinética , Meia-Vida , Taxa de Depuração Metabólica , Estrutura Molecular , Ratos , Ratos Sprague-Dawley , Relação Estrutura-AtividadeRESUMO
A series of substituted tetrahydrofuroyl-1-phenylalanine derivatives was prepared and evaluated as VLA-4 antagonists. Substitution of the alpha carbon of the tetrahydrofuran with aryl groups increased the specificity for VLA-4 versus alpha(4)beta(7) while amide substitution increased the potency of the series without increasing the specificity. Substitution of the beta carbon of the tetrahydrofuran with keto or amino groups slightly improved the specificity for VLA-4 versus alpha(4)beta(7) but with a significant loss in binding affinity for VLA-4.
Assuntos
Integrina alfa4beta1/antagonistas & inibidores , Fenilalanina/análogos & derivados , Sulfonamidas/síntese química , Sulfonamidas/farmacocinética , Administração Oral , Animais , Furanos/síntese química , Furanos/química , Furanos/farmacocinética , Furanos/farmacologia , Humanos , Concentração Inibidora 50 , Ligantes , Peso Molecular , Fenilalanina/síntese química , Fenilalanina/farmacocinética , Fenilalanina/farmacologia , Ratos , Estereoisomerismo , Relação Estrutura-Atividade , Especificidade por Substrato , Sulfonamidas/sangue , Sulfonamidas/química , Molécula 1 de Adesão de Célula Vascular/metabolismoRESUMO
N-(3,5-Dichlorophenylsulfonyl)-(R)-thioprolyl biarylalanine 10a has been identified as a potent and specific antagonist of the alpha(4)beta(1) integrin. Altering the configuration of thioproline from R to S led to a series of dual antagonists of alpha(4)beta(1) and alpha(4)beta(7), and the N-acetyl analogue 8b was found to be the most potent dual antagonist. A binding site model for alpha(4)beta(1) and alpha(4)beta(7) is proposed to explain the structure-activity relationship.
Assuntos
Alanina/farmacologia , Integrinas/antagonistas & inibidores , Receptores de Retorno de Linfócitos/antagonistas & inibidores , Compostos de Sulfidrila/farmacologia , Alanina/análogos & derivados , Alanina/química , Integrina alfa4beta1 , Relação Estrutura-Atividade , Compostos de Sulfidrila/químicaRESUMO
The alpha(4)beta(1) and alpha(4)beta(7) integrins are implicated in several inflammatory disease states. Systematic SAR studies of an alpha(4)beta(1)-specific arylsulfonyl-Pro-Tyr lead led to the identification of a new alpha(4)beta(7) binding site, best captured by O-carbamates of Tyr for this structural class. Several compounds showed a 200- to 400-fold improvement in alpha(4)beta(7) binding affinity while maintaining subnanomolar alpha(4)beta(1) activity, for example 2l, VCAM-Ig alpha(4)beta(1) IC(50)=0.13 nM, VCAM-Ig alpha(4)beta(7) IC(50)=1.92 nM.
Assuntos
Carbamatos/farmacologia , Integrinas/antagonistas & inibidores , Receptores de Retorno de Linfócitos/antagonistas & inibidores , Carbamatos/química , Carbamatos/farmacocinética , Humanos , Integrina alfa4beta1 , Células Jurkat , Relação Estrutura-AtividadeRESUMO
VLA-4 (alpha(4)beta(1), very late activating antigen-4), a key cell surface integrin plays an important role in inflammation by promoting leukocyte attachment and extravasation from the vasculature into the peripheral tissues. As such, VLA-4 antagonists may be useful in the treatment, prevention, and suppression of diseases where cell adhesion and migration are important such as asthma, rheumatoid arthritis, and multiple sclerosis. Herein, we report on the discovery, synthesis, and biological evaluation of amidines as small molecule antagonists of VLA-4.
Assuntos
Amidas/química , Amidinas/química , Integrina alfa4beta1/antagonistas & inibidores , Área Sob a CurvaRESUMO
A series of substituted N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl- and (L)-azetidyl-beta-biaryl beta-alanine derivatives was prepared as selective and potent VLA-4 antagonists. The 2,6-dioxygenated biaryl substitution pattern is important for optimizing potency. Oral bioavailability was variable and may be a result of binding to circulating plasma proteins.
Assuntos
Integrina alfa4beta1/antagonistas & inibidores , Propionatos/síntese química , Administração Oral , Alanina/síntese química , Alanina/farmacocinética , Alanina/farmacologia , Disponibilidade Biológica , Humanos , Concentração Inibidora 50 , Células Jurkat , Taxa de Depuração Metabólica , Propionatos/farmacocinética , Propionatos/farmacologia , Ligação Proteica , Relação Estrutura-Atividade , Molécula 1 de Adesão de Célula Vascular/metabolismoRESUMO
The alpha(4) integrin, alpha(4)beta(7), plays an important role in recruiting circulating lymphocytes to the gastrointestinal tract, where its ligand mucosal addressin cell adhesion molecule-1 (MAdCAM-1) is preferentially expressed on high endothelial venules (HEVs). Dual antagonists of alpha(4)beta(1) and alpha(4)beta(7), N-(2,6-dichlorobenzoyl)-(L)-4-(2',6'-bis-methoxyphenyl)phenylalanine (TR14035) and N-(N-[(3,5-dichlorobenzene)sulfonyl]-2-(R)-methylpropyl)-(D)-phenylalanine (compound 1), were tested for their ability to block the binding of alpha(4)beta(7)-expressing cells to soluble ligand in suspension and under in vitro and in vivo shear flow. Compound 1 and TR14035 blocked the binding of human alpha(4)beta(7) to an (125)I-MAdCAM-Ig fusion protein with IC(50) values of 2.93 and 0.75 nM, respectively. Both compounds inhibited binding of soluble ligands to alpha(4)beta(1) or alpha(4)beta(7) on cells of human or rodent origin with similar potency. Under shear flow in vitro, TR14035 and compound 1 blocked binding of human alpha(4)beta(7)-expressing RPMI-8866 cells or murine mesenteric lymph node lymphocytes to MAdCAM-Ig with IC(50) values of 0.1 and 1 microM, respectively. Intravital microscopy was used to quantitate alpha(4)-dependent adhesion of fluorescent murine lymphocytes in Peyer's patch HEVs. When cells were prestimulated with 2 mM Mn(2+) to activate alpha(4)beta(7) binding to ligand, anti-alpha(4) monoclonal antibody (mAb) [10 mg/kg (mpk) i.v.] blocked adhesion by 95%, and anti-beta(1) mAb did not block adhesion, demonstrating that this interaction was dependent on alpha(4)beta(7). TR14035 blocked adhesion to HEVs [ED(50) of 0.01-0.1 mpk i.v.], and compound 1 blocked adhesion by 47% at 10 mpk i.v. Thus, alpha(4)beta(7)/alpha(4)beta(1) antagonists blocked alpha(4)beta(7)-dependent adhesion of lymphocytes to HEVs under both in vitro and in vivo shear flow.
Assuntos
Integrinas/antagonistas & inibidores , Fenilalanina/farmacologia , Receptores de Retorno de Linfócitos/antagonistas & inibidores , Animais , Anticorpos Bloqueadores/farmacologia , Anticorpos Monoclonais/farmacologia , Protocolos de Quimioterapia Combinada Antineoplásica/imunologia , Adesão Celular/efeitos dos fármacos , Moléculas de Adesão Celular , Ciclofosfamida/imunologia , Doxorrubicina/imunologia , Endotélio Vascular/citologia , Endotélio Vascular/efeitos dos fármacos , Etoposídeo/imunologia , Feminino , Citometria de Fluxo , Humanos , Imunoglobulina G/isolamento & purificação , Imunoglobulina G/farmacologia , Imunoglobulinas , Integrina alfa4beta1 , Ligantes , Linfócitos/efeitos dos fármacos , Metotrexato/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Mucoproteínas/antagonistas & inibidores , Nódulos Linfáticos Agregados/citologia , Nódulos Linfáticos Agregados/efeitos dos fármacos , Fenilalanina/análogos & derivados , Proteínas Recombinantes de Fusão/farmacologia , ReologiaRESUMO
Given the proposed involvement of VLA-4 in inflammatory processes, a program to identify orally active VLA-4 antagonists was initiated. Herein, we report the discovery of a N-tetrahydrofuroyl-(L)-phenylalanine derivative (17) and related analogues as potent VLA-4 antagonists with good oral bioavailability.
Assuntos
Integrina alfa4beta1/antagonistas & inibidores , Fenilalanina/análogos & derivados , Administração Oral , Animais , Sítios de Ligação , Disponibilidade Biológica , Relação Dose-Resposta a Droga , Furanos/síntese química , Furanos/química , Furanos/farmacocinética , Furanos/farmacologia , Meia-Vida , Concentração Inibidora 50 , Macaca mulatta , Fenilalanina/síntese química , Fenilalanina/farmacocinética , Fenilalanina/farmacologia , Ratos , Ratos Sprague-Dawley , Relação Estrutura-Atividade , Sulfonamidas/síntese química , Sulfonamidas/química , Sulfonamidas/farmacocinética , Sulfonamidas/farmacologia , Molécula 1 de Adesão de Célula Vascular/metabolismoRESUMO
A series of substituted N-(3,5-dichlorobenzenesulfonyl)-L-prolyl- and alpha-methyl-L-prolyl-phenylalanine derivatives was prepared as VLA-4/VCAM antagonists. The compounds showed excellent potency with a wide variety of neutral, polar, electron withdrawing or donating groups on the phenylalanine ring (IC50 approximately 1 nM). Heteroaryl ring substitution for phenylalanine was also well tolerated. Pharmacokinetic studies in rat were performed on a representative set of compounds in both series.
Assuntos
Dipeptídeos/farmacocinética , Integrina alfa4beta1/antagonistas & inibidores , Animais , Disponibilidade Biológica , Dipeptídeos/síntese química , Dipeptídeos/química , Cães , Haplorrinos , Concentração Inibidora 50 , Taxa de Depuração Metabólica , Fenilalanina , Ratos , Ratos Sprague-Dawley , Ovinos , Relação Estrutura-Atividade , Sulfonas , Molécula 1 de Adesão de Célula Vascular/efeitos dos fármacosRESUMO
An alpha4beta1/alpha4beta7 dual antagonist, 35S-compound 1, was used as a model ligand to study the effect of divalent cations on the activation state and ligand binding properties of alpha4 integrins. In the presence of 1 mM each Ca2+/Mg2+, 35S-compound 1 bound to several cell lines expressing both alpha4beta1 and alpha4beta7, but 2S-[(1-benzenesulfonyl-pyrrolidine-2S-carbonyl)-amino]-4-[4-methyl-2S-(methyl-[2-[4-(3-o-tolyl-ureido)-phenyl]-acetyl]-amino) pentanoylamino]-butyric acid (BIO7662), a specific alpha4beta1 antagonist, completely inhibited 35S-compound 1 binding, suggesting that alpha4beta1 was responsible for the observed binding. 35S-Compound 1 bound RPMI-8866 cells expressing predominantly alpha4beta7 with a KD of 1.9 nM in the presence of 1 mM Mn2+, and binding was inhibited only 29% by BIO7662, suggesting that the probe is a potent antagonist of activated alpha4beta7. With Ca2+/Mg2+, 35S-compound 1 bound Jurkat cells expressing primarily alpha4beta1 with a KD of 18 nM. In contrast, the binding of 35S-compound 1 to Mn2+-activated Jurkat cells occurred slowly, reaching equilibrium by 60 min, and failed to dissociate within another 60 min. The ability of four alpha4beta1/alpha4beta7 antagonists to block binding of activated alpha4beta1 or alpha4beta7 to vascular cell adhesion molecule-1 or mucosal addressin cell adhesion molecule-1, respectively, or to 35S-compound 1 was measured, and a similar rank order of potency was observed for native ligand and probe. Inhibition of 35S-compound 1 binding to alpha4beta1 in Ca2+/Mg2+ was used to identify nonselective antagonists among these four. These studies demonstrate that alpha4beta1 and alpha4beta7 have distinct binding properties for the same ligand, and binding parameters are dependent on the state of integrin activation in response to different divalent cations.
Assuntos
Cátions Bivalentes/metabolismo , Dipeptídeos/farmacologia , Integrina alfa4beta1/antagonistas & inibidores , Integrinas/antagonistas & inibidores , Fenilalanina/farmacologia , Compostos de Fenilureia/farmacologia , Sítios de Ligação , Linhagem Celular , Dipeptídeos/química , Humanos , Integrina alfa4beta1/metabolismo , Integrinas/metabolismo , Células Jurkat , Células K562 , Cinética , Ligantes , Fenilalanina/análogos & derivados , Fenilalanina/química , Compostos de Fenilureia/química , Ligação Proteica , Ensaio Radioligante , Radioisótopos de Enxofre , Células Tumorais Cultivadas , Molécula 1 de Adesão de Célula Vascular/metabolismoRESUMO
Acylated beta-amino acids are described as potent, specific and orally bioavailable antagonists of VLA-4. The initial lead was identified from a combinatorial library. Subsequent optimization using a traditional medicinal chemistry approach led to significant improvement in potency (up to 8-fold) while maintaining good pharmacokinetic properties.