RESUMEN
We describe the synthesis, MMP-2 and 9 potency, and in vitro evaluation of a series of α-sulfone hydroxmate MMP inhibitors conjugated to a series of dyes with different absorption/emission lamina maxima's that can be used to visualize tumors.
Asunto(s)
Diseño de Fármacos , Colorantes Fluorescentes/química , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Inhibidores de la Metaloproteinasa de la Matriz/síntesis química , Humanos , Células MCF-7 , Metaloproteinasa 2 de la Matriz/química , Metaloproteinasa 9 de la Matriz/química , Inhibidores de la Metaloproteinasa de la Matriz/química , Inhibidores de la Metaloproteinasa de la Matriz/metabolismo , Microscopía ConfocalRESUMEN
The integrin alpha(v)beta(3), vitronectin receptor, is expressed in a number of cell types and has been shown to mediate adhesion of osteoclasts to bone matrix, vascular smooth muscle cell migration, and angiogenesis. We recently disclosed the discovery of a tripeptide Arg-Gly-Asp (RGD) mimic, which has been shown to be a potent inhibitor of the integrin alpha(v)beta(3) and has excellent anti-angiogenic properties including its suppression of tumor growth in animal models. In other investigations involving RGD mimics, only compounds containing the S-isomers of the beta-amino acids have been shown to be potent. We were surprised to find the potencies of analogs containing enantiomerically pure S-isomers of beta-amino acids which were only marginally better than the corresponding racemic mixtures. We therefore synthesized RGD mimics containing R-isomers of beta-amino acids and found them to be relatively potent inhibitors of alpha(v)beta(3). One of the compounds was examined in tumor models in mice and has been shown to significantly reduce the rate of growth and the size of tumors.
Asunto(s)
Aminoácidos/química , Antineoplásicos/química , Antineoplásicos/farmacología , Integrina alfaVbeta3/antagonistas & inhibidores , Imitación Molecular , Oligopéptidos/química , Oligopéptidos/farmacología , Aminoácidos/síntesis química , Animales , Antineoplásicos/farmacocinética , Neoplasias del Colon , Hipercalcemia/inducido químicamente , Isomerismo , Melanoma , Ratones , Ratones Endogámicos , Oligopéptidos/farmacocinética , Neoplasias Cutáneas , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
The integrin alpha(v)beta(3) is expressed in a number of cell types and is thought to play a major role in several pathological conditions. Various small molecules that inhibit the integrin have been shown to suppress tumor growth and retinal angiogenesis. The tripeptide Arg-Gly-Asp (RGD), a common binding motif in several ligands that bind to alpha(v)beta(3), has been depeptidized and optimized in our efforts toward discovering a small molecule inhibitor. We recently disclosed the synthesis and biological activity of several small molecules that did not contain any peptide bond and mimic the tripeptide RGD. The phenethyl group in one of the lead compounds was successfully replaced with a cyclopropyl moiety. The new lead compound was optimized for potency, selectivity, and for its ADME properties. We describe herein the discovery, synthesis, and optimization of cyclopropyl containing analogs that are potent and selective inhibitors of alpha(v)beta(3).
Asunto(s)
Acetatos/síntesis química , Acetatos/farmacología , Integrina alfaVbeta3/antagonistas & inhibidores , Naftiridinas/síntesis química , Naftiridinas/farmacología , Animales , Área Bajo la Curva , Línea Celular , Cromatografía Líquida de Alta Presión , Cristalografía por Rayos X , Diseño de Fármacos , Semivida , Humanos , Indicadores y Reactivos , Masculino , Ratones , Ratas , Ratas Sprague-Dawley , Espectrofotometría Ultravioleta , Relación Estructura-Actividad , TransfecciónRESUMEN
We describe a series of pyrazole and isoxazole analogs as antagonists of the alpha(v)beta3 receptor. Compounds showed low to sub-nanomolar potency against alpha(v)beta3, as well as good selectivity against alpha(IIb)beta3. In HT29 cells, most analogs also demonstrated significant selectivity against alpha(v)beta6. Several compounds showed good pharmacokinetic properties in rats, in addition to anti-angiogenic activity in a mouse corneal micropocket model. Compounds were synthesized in a straightforward manner from readily available glutarate precursors.
Asunto(s)
Integrina alfaVbeta3/antagonistas & inhibidores , Isoxazoles/síntesis química , Isoxazoles/farmacología , Pirazoles/síntesis química , Pirazoles/farmacología , Animales , Línea Celular , Humanos , Integrina alfaVbeta3/metabolismo , Isoxazoles/química , Isoxazoles/farmacocinética , Ratones , Estructura Molecular , Pirazoles/química , Pirazoles/farmacocinética , Ratas , Relación Estructura-ActividadRESUMEN
We describe a series of 1,2,4-oxadiazoles, which are potent antagonists of the integrin alpha(v)beta3 and, in addition, show selectivity relative to the other beta3 integrin alpha(IIb)beta3. In whole cells, the majority of these analogs also demonstrated modest selectivity against other alpha(v) integrins such as alpha(v)beta1 and alpha(v)beta6.
Asunto(s)
Butiratos/síntesis química , Butiratos/farmacología , Integrina alfaVbeta3/antagonistas & inhibidores , Oxadiazoles/síntesis química , Oxadiazoles/farmacología , Antígenos de Neoplasias , Butiratos/química , Línea Celular , Humanos , Integrinas/antagonistas & inhibidores , Estructura Molecular , Oxadiazoles/química , Receptores de Vitronectina/antagonistas & inhibidores , Relación Estructura-ActividadRESUMEN
We describe a series of 2,5 thiazole containing compounds, which are potent antagonists of the integrin alpha(v)beta3 and show selectivity relative to the other integrins, such as alpha(IIb)beta3 and alpha(v)beta6. These analogs were demonstrated to have high bioavailability relative to other relative heterocyclic analogs.
Asunto(s)
Butiratos/síntesis química , Butiratos/farmacocinética , Integrina alfaVbeta3/antagonistas & inhibidores , Tiazoles/síntesis química , Tiazoles/farmacocinética , Administración Oral , Animales , Antígenos de Neoplasias , Disponibilidad Biológica , Butiratos/administración & dosificación , Perros , Evaluación Preclínica de Medicamentos , Haplorrinos , Integrinas/antagonistas & inhibidores , Estructura Molecular , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/antagonistas & inhibidores , Ratas , Relación Estructura-Actividad , Tiazoles/administración & dosificaciónRESUMEN
Integrins expressed on endothelial cells modulate cell migration and survival during angiogenesis. Integrins expressed on carcinoma cells potentiate metastasis by facilitating invasion and movement across blood vessels. We describe the activities of two synthetic low-molecular-weight peptidomimetics of the ligand amino acid sequence arg-gly-asp (RGD) in integrin-based functional assays in vitro. We also evaluate efficacy and potential mechanisms of action in models of both spontaneous and experimental metastasis. Broad-spectrum potency against the family of alpha v subunit-containing integrins was observed, with significantly less potency against alpha5beta1 and alpha(IIb)beta3. Both endothelial and tumor cell migration mediated by alpha(v)beta3 was inhibited, whereas proliferation of endothelial cells but not tumor cells was diminished. Continuous infusion of compound by minipumps or oral administration twice daily significantly reduced metastatic tumor burden in the lungs of mice despite no reduction in growth of 435/HAL primary tumors, and only a slight reduction in tumor cells detected in circulating blood. Delaying treatment in this model until after extensive dissemination of tumor cells to the lungs had occurred, and after primary tumor resection, still produced significant efficacy. Conversely, administration of the agent for only the first 18 h after tumor-cell inoculation into the tail vein also resulted in decreased metastases observed after several weeks. These data suggest these compounds or their relatives have potential to interfere with both early and late steps of metastasis involving tumor and endothelial cell functions. Furthermore, the metastatic process can be effectively inhibited independently of primary tumor growth using integrin antagonists.
Asunto(s)
Antineoplásicos/uso terapéutico , Neoplasias de la Mama/patología , Carcinoma Ductal/secundario , Dipéptidos/uso terapéutico , Integrina alfaVbeta3/antagonistas & inhibidores , Neoplasias Pulmonares/secundario , Proteínas de Neoplasias/antagonistas & inhibidores , Compuestos Orgánicos/uso terapéutico , Pirimidinas/uso terapéutico , Administración Oral , Animales , Antineoplásicos/administración & dosificación , Antineoplásicos/farmacología , Carcinoma Ductal/tratamiento farmacológico , Carcinoma Ductal/patología , Carcinoma Ductal/prevención & control , Carcinoma Ductal/cirugía , División Celular/efectos de los fármacos , Línea Celular Tumoral/trasplante , Movimiento Celular/efectos de los fármacos , Neoplasias del Colon/patología , Dipéptidos/administración & dosificación , Dipéptidos/farmacología , Esquema de Medicación , Células Endoteliales/citología , Células Endoteliales/efectos de los fármacos , Endotelio Vascular/citología , Femenino , Humanos , Bombas de Infusión Implantables , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/prevención & control , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones SCID , Células Neoplásicas Circulantes , Oligopéptidos , Compuestos Orgánicos/administración & dosificación , Compuestos Orgánicos/farmacología , Pirimidinas/administración & dosificación , Pirimidinas/farmacología , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Members of the integrin family influence several aspects of tumor progression and metastasis, including cell survival, proliferation, and angiogenesis. Specific integrins such as alpha(v)beta(3) and alpha(v)beta(5) are involved in regulating endothelial cell function, and thus angiogenesis. We evaluated the effect of the alpha(v)beta(3)/alpha(v)beta(5) integrin antagonist S247 on the growth and angiogenesis of colon cancer liver metastases in an orthotopic murine model. Murine colon cancer cells were injected into the spleens of BALB/c mice to produce liver metastases. On day 7, miniature osmotic pumps were implanted into the subcutis to continuously infuse either saline or 70 mg/kg/day S247. All mice were sacrificed when control mice became moribund. Mice that received S247 developed significantly fewer liver metastases than did controls (P < 0.05). Using the same model, a subsequent survival study was performed. Mice were sacrificed when moribund as determined by an observer blinded to the treatment given. Treatment with S247 significantly prolonged overall survival (P < 0.05). Interestingly, primary tumors in the spleen were the cause of death in the S247-treated group as S247 appeared to have little effect on these tumors. Immunohistochemical staining demonstrated a significant reduction of vessels in liver metastases of S247-treated mice (P < 0.001), a significant increase in endothelial cell apoptosis (P < 0.05), and a significant decrease in pericyte coverage (P < 0.0001). To determine the role of S247 on angiogenesis, we examined the effect of S247 in vitro on human umbilical vein endothelial cells (HUVECs) and human vascular smooth muscle cells (hVSMCs). The addition of S247 to HUVECs and hVSMCs growing on vitronectin-coated flasks and in Matrigel significantly impaired cell growth and colony formation, respectively (P < 0.05). Furthermore, S247 completely inhibited the attachment of HUVECs and hVSMCs and increased apoptosis by six- to 9fold compared with controls. In in vitro invasion assays, S247-treated cells demonstrated decreased migration (P < 0.05). In conclusion, S247 demonstrated significant antimetastatic and antiangiogenic activity and impaired both endothelial and hVSMC/pericyte function in vitro and in vivo. The use of agents such as integrin antagonists that target multiple cell types involved in angiogenesis may be a more effective method of inhibiting angiogenesis than agents targeting only the endothelial cells.