RESUMO
Geographic atrophy (GA) is an advanced form of age-related macular degeneration (AMD) characterized by atrophy of the retinal pigment epithelium (RPE), loss of photoreceptors, and disruption of choriocapillaris. Excessive light exposure is toxic to the retina and is a known risk factor for AMD. We first investigated the effects of blue light-induced phototoxicity on RPE and photoreceptors in nonhuman primates (NHPs, a model of progressive retinal degeneration) and then evaluated the potential cyto- and neuroprotective effects of the brimonidine drug delivery system (Brimo DDS). In the first set of experiments related to model development, parafoveal lesions of varying severity were induced using blue light irradiation of the retina of cynomolgus monkeys to evaluate the level of phototoxicity in the RPE and photoreceptors. RPE damage was assessed using fundus autofluorescence imaging to quantify areas of hypofluorescence, while thinning of the outer nuclear layer (ONL, photoreceptor nuclei) was quantified using optical coherence tomography (OCT). Photoreceptor function was assessed using multifocal electroretinography (mfERG). RPE damage progressively increased across all lesion severities from 2 to 12 weeks, as did the extent of ONL thinning. Lesions of high severity continued to show reduction in mfERG amplitude, reaching a statistically significant maximum reduction at 12 weeks. Collectively, the first set of experiments showed that blue light irradiation of the NHP eye resulted in progressive retinal degeneration identified by damage to RPE, ONL thinning, and disrupted photoreceptor function - hallmarks of GA in humans. We then used the model to evaluate the cyto- and neuroprotective effects of Brimo DDS, administered as a therapeutic after allowing the lesions to develop for 5 weeks. Placebo DDS or Brimo DDS were administered intravitreally and a set of untreated animals were used as an additional control. In the placebo DDS group, hypofluorescence area continued to increase from baseline, indicating progressive RPE damage, while progression was significantly slowed in eyes receiving Brimo DDS. Likewise, ONL thinning continued to progress over time in eyes that received the placebo DDS, but was reduced in Brimo DDS-treated eyes. Pharmacologically relevant brimonidine concentrations were sustained in the retina for up to 26 weeks following Brimo DDS administration. In summary, Brimo DDS demonstrated cyto- and neuroprotective effects in a novel NHP GA model of progressive retinal degeneration.
Assuntos
Tartarato de Brimonidina/administração & dosagem , Corioide/diagnóstico por imagem , Citoproteção/efeitos dos fármacos , Sistemas de Liberação de Medicamentos , Atrofia Geográfica/tratamento farmacológico , Neuroproteção/efeitos dos fármacos , Segmento Externo das Células Fotorreceptoras da Retina/patologia , Agonistas de Receptores Adrenérgicos alfa 2/administração & dosagem , Animais , Corioide/efeitos dos fármacos , Corioide/efeitos da radiação , Modelos Animais de Doenças , Eletrorretinografia , Angiofluoresceinografia/métodos , Fundo de Olho , Atrofia Geográfica/diagnóstico , Macaca fascicularis , Soluções Oftálmicas/administração & dosagem , Segmento Externo das Células Fotorreceptoras da Retina/efeitos dos fármacos , Segmento Externo das Células Fotorreceptoras da Retina/efeitos da radiação , Epitélio Pigmentado da Retina/efeitos dos fármacos , Epitélio Pigmentado da Retina/patologia , Epitélio Pigmentado da Retina/efeitos da radiação , Tomografia de Coerência Óptica/métodos , Acuidade VisualRESUMO
PURPOSE: To compare the durability of Kenalog, Trivaris, Triesence, and compounding pharmacy preservative-free triamcinolone acetonide in pigmented rabbits with syneretic vitreous using direct visualization, pharmacodynamics, and pharmacokinetics. METHODS: Twenty-five Dutch-belted rabbits were used. Pharmacokinetic experiment: Rabbits were intravitreally injected with one of four 4-mg triamcinolone acetonide formulations. Wide-field imaging was serially performed to document residual drug mass. Pharmacodynamics experiment: Four triamcinolone acetonide groups and one control group received intravitreal recombinant human vascular endothelial growth factor 165 every 2 weeks and were followed with fluorescein angiography to assess vascular endothelial growth factor retinal vasculopathy as a measure of residual steroid effect. Particle size of the formulations was measured with Mastersizer 2000. RESULTS: Remaining triamcinolone acetonide mass after 19 weeks: 12,091 ± 2,512 pixels for the Kenalog group, 1,307.36 ± 695.57 for Trivaris, 5577 ± 1477 for Triesence, and 1,535 ± 329 for compounded preservative-free triamcinolone acetonide. Kenalog suppressed recombinant human vascular endothelial growth factor-induced retinopathy more effectively than the other triamcinolone acetonide groups at Week 39, the final time point assessed. Particle size (90th percentile) was 47 µm for Kenalog, 26 µm for Triesence, and 22 µm for both compounded preservative-free triamcinolone acetonide and Trivaris. CONCLUSION: Triamcinolone acetonide formulations do not have the same pharmacokinetics/pharmacodynamics. Kenalog has the longest vitreous visibility and durability. Particle size appears to correlate with efficacy and durability.
Assuntos
Glucocorticoides/farmacologia , Glucocorticoides/farmacocinética , Neovascularização Retiniana/metabolismo , Triancinolona Acetonida/farmacologia , Triancinolona Acetonida/farmacocinética , Corpo Vítreo/metabolismo , Animais , Disponibilidade Biológica , Composição de Medicamentos , Angiofluoresceinografia , Glucocorticoides/química , Meia-Vida , Injeções Intravítreas , Tamanho da Partícula , Conservantes Farmacêuticos , Coelhos , Proteínas Recombinantes , Neovascularização Retiniana/induzido quimicamente , Neovascularização Retiniana/prevenção & controle , Solubilidade , Triancinolona Acetonida/química , Fator A de Crescimento do Endotélio Vascular/sangue , Fator A de Crescimento do Endotélio Vascular/toxicidadeRESUMO
Purpose: To assess the intraocular pressure (IOP)-lowering effect of a biodegradable bimatoprost implant following selective laser trabeculoplasty (SLT) in a canine model. Methods: Unilateral SLT was performed in 11 normotensive, treatment-naive beagle dogs. IOP was measured at baseline (pre-SLT) and weekly post-SLT (≤10 weeks). After IOP returned to baseline or at 10 weeks (whichever occurred first), a sustained-release bimatoprost implant was administered bilaterally in the anterior chamber of each animal. IOP was measured weekly for 4 weeks and then every 2 weeks up to week 42. Results: The main outcomes included the IOP change (%) from baseline, calculated in both eyes in the overall population, SLT responder subgroup (defined by peak IOP reduction from baseline ≥3 mmHg or ≥15% for >1 week post-SLT), and SLT nonresponder subgroup (defined by peak IOP reduction from baseline <3 mmHg or <15%). The bimatoprost implant lowered IOP similarly in both the SLT-treated and fellow SLT-naive eyes. Following bimatoprost implant administration, the mean (standard deviation [SD]) peak IOP reduction from baseline was 34.4% (8.5%) in SLT-treated eyes and 35.7% (5.9%) in fellow SLT-naive eyes. The bimatoprost implant lowered IOP comparably (P > 0.17) in eyes that responded to SLT (mean [SD] peak IOP reduction, 34.6% [10.7%]; n = 6) and those that did not (mean [SD] peak IOP reduction, 34.1% [6.1%]; n = 5). Conclusion: The bimatoprost implant effectively lowered IOP in eyes pretreated with SLT, regardless of response to SLT. The current data suggest that eyes previously treated with SLT can still benefit from the intracameral bimatoprost implant.
Assuntos
Terapia a Laser , Hipertensão Ocular , Trabeculectomia , Animais , Bimatoprost/farmacologia , Bimatoprost/uso terapêutico , Preparações de Ação Retardada , Cães , Pressão Intraocular , Lasers , Resultado do TratamentoRESUMO
PURPOSE: To assess the efficacy of a dexamethasone (DEX) intravitreal implant in a rabbit model of anterior and intermediate uveitis. METHODS: Experimental anterior and intermediate uveitis was induced by a unilateral intracameral injection of Mycobacterium tuberculosis H37Ra antigen in preimmunized rabbits. Four days after uveitis induction, rabbits received DEX implant or underwent a sham procedure (no implant). Clinical and histopathologic signs of uveitis were assessed for 13 days, and levels of inflammatory markers in the iris/ciliary body were measured after 21 days. RESULTS: All signs of anterior and intermediate uveitis were reduced by the DEX implant compared with sham procedure. At day 13, mean anterior chamber cell scores ± SD for the DEX implant versus the sham procedure were, respectively, 1.9 ± 1.3 versus 4.0 ± 0.0 (P = 0.04), and mean total histologic inflammatory scores were 3.9 ± 2.5 versus 15.4 ± 6.0 (P = 0.026). Similarly, at day 13, mean vitreous haze severity scores (SD) for the DEX implant versus the sham procedure were, respectively, 0.1 ± 0.2 versus 2.7 ± 1.5 (P = 0.026), and mean vitreous inflammatory cell infiltration scores were 0.0 ± 0.0 versus 1.5 ± 1.3. Treatment with the DEX intravitreal implant also significantly reduced the proinflammatory immune response, as measured by cytokine levels in iris/ciliary body. CONCLUSIONS: A single administration of DEX implant significantly reduced inflammation in an animal model of anterior and intermediate uveitis.
Assuntos
Dexametasona/administração & dosagem , Modelos Animais de Doenças , Glucocorticoides/administração & dosagem , Tuberculose Ocular/tratamento farmacológico , Uveíte Anterior/tratamento farmacológico , Uveíte Intermediária/tratamento farmacológico , Animais , Antígenos de Bactérias/toxicidade , Biomarcadores/metabolismo , Corpo Ciliar/metabolismo , Citocinas/metabolismo , Implantes de Medicamento , Iris/metabolismo , Mycobacterium tuberculosis/imunologia , Coelhos , Resultado do Tratamento , Tuberculose Ocular/diagnóstico , Tuberculose Ocular/metabolismo , Tuberculose Ocular/microbiologia , Uveíte Anterior/diagnóstico , Uveíte Anterior/metabolismo , Uveíte Anterior/microbiologia , Uveíte Intermediária/diagnóstico , Uveíte Intermediária/metabolismo , Uveíte Intermediária/microbiologia , Corpo VítreoRESUMO
PURPOSE: To evaluate dexamethasone pharmacokinetics after implantation of a sustained-release dexamethasone (DEX) intravitreal implant in nonvitrectomized and vitrectomized eyes. METHODS: The right eyes of 25 rabbits underwent vitrectomy; contralateral eyes served as nonvitrectomy controls. The 0.7-mg DEX implant was injected into both eyes, and drug concentrations were determined in the vitreous humor and retina for 31 days (on days 2, 8, 15, 22, and 31). RESULTS: DEX was present in nonvitrectomized and vitrectomized eyes for at least 31 days. There were no statistically significant differences in DEX concentration between nonvitrectomized and vitrectomized eyes at any time point (P > 0.05). The maximum concentration of DEX in nonvitrectomized versus vitrectomized eyes for vitreous humor was 791 ng/mL (day 22) versus 731 ng/mL (day 22), respectively, and for retina it was 4110 ng/mL (day 15) versus 3670 ng/mL (day 22), respectively. Mean absorption (AUC(0-tlast)) of dexamethasone in nonvitrectomized and vitrectomized eyes was not different for both the vitreous humor (13,600 vs. 15,000 ng/day/mL; P = 0.73) and retina (67,600 vs. 50,200 ng/day/mL; P = 0.47). CONCLUSIONS: The vitreoretinal pharmacokinetic profiles were similar between nonvitrectomized and vitrectomized eyes. These observations are consistent with clinical findings of the DEX implant in patients who have undergone vitrectomy and should reduce concerns about the use of the DEX implant in eyes that have undergone vitrectomy.
Assuntos
Dexametasona/farmacocinética , Oftalmopatias/tratamento farmacológico , Vitrectomia , Corpo Vítreo/metabolismo , Animais , Preparações de Ação Retardada/administração & dosagem , Preparações de Ação Retardada/farmacocinética , Dexametasona/administração & dosagem , Modelos Animais de Doenças , Implantes de Medicamento , Oftalmopatias/metabolismo , Oftalmopatias/cirurgia , Seguimentos , Glucocorticoides/administração & dosagem , Glucocorticoides/farmacocinética , Masculino , Taxa de Depuração Metabólica , Segmento Posterior do Olho , Coelhos , Corpo Vítreo/efeitos dos fármacos , Corpo Vítreo/cirurgiaRESUMO
Purpose. Pars plana vitrectomy (PPV) has been reported to reduce macular thickness and improve visual acuity in patients with diabetic macular edema (ME). The hypothesis for the study was that after PPV, clearance is accelerated and VEGF concentrations are reduced. To test this hypothesis, hVEGF(165) injections were performed in rabbit eyes, with and without PPV, and vitreous VEGF levels were measured as a function of time. Methods. The PPV group rabbits had a bilateral 25-gauge PPV, and in the no-PPV group, rabbits had intact vitreous. Intravitreal injections of hVEGF(165) were performed, and the animals were euthanatized at time points up to 7 days. The vitreous was isolated and an enzyme-linked immunosorbent assay was used to measure the VEGF levels. Pharmacokinetic parameters were determined in a noncompartmental analysis approach. Results. Mean vitreous VEGF levels decreased more rapidly in eyes subjected to PPV than in no-PPV eyes. The vitreous VEGF half-life (t([)(1/2)(])) in PPV eyes was 10 times shorter than that in normal eyes. In addition, mean clearance and mean area under the curve (AUC) increased and decreased, respectively, in eyes that underwent PPV. Conclusions. VEGF clearance is increased after PPV. Reducing VEGF concentrations in the vitreous post-PPV may partially explain the improvement in macular thickness in some patients with ME. Unexpectedly, the half-life of VEGF in the vitreous, even in no-PPV eyes, was <3 hours, whereas compounds of similar molecular weight typically have longer vitreous half-lives. The back of the eye may be uniquely adapted with rapid-clearance mechanisms to regulate vitreous VEGF levels. Further study is suggested.
Assuntos
Fator A de Crescimento do Endotélio Vascular/farmacocinética , Vitrectomia , Corpo Vítreo/metabolismo , Animais , Área Sob a Curva , Ensaio de Imunoadsorção Enzimática , Meia-Vida , CoelhosRESUMO
Beta-catenin is a component of stable cell adherent complexes whereas its free form functions as a transcription factor that regulate genes involved in oncogenesis and metastasis. Free beta-catenin is eliminated by two adenomatous polyposis coli (APC)-dependent proteasomal degradation pathways regulated by glycogen synthase kinase 3beta (GSK3 beta) or p53-inducible Siah-1. Dysregulation of beta-catenin turnover consequent to mutations in critical genes of the APC-dependent pathways is implicated in cancers such as colorectal cancer. We have identified a novel retinoid X receptor (RXR)-mediated APC-independent pathway in the regulation of beta-catenin. In this proteasomal pathway, RXR agonists induce degradation of beta-catenin and RXR alpha and repress beta-catenin-mediated transcription. In vivo, beta-catenin interacts with RXR alpha in the absence of ligand, but RXR agonists enhanced the interaction. RXR agonist action was not impaired by GSK3 beta inhibitors or deletion of the GSK3 beta-targeted sequence from beta-catenin. In APC- and p53-mutated colorectal cancer cells, RXR agonists still inactivated endogenous beta-catenin via RXR alpha. Interestingly, deletion of the RXR alpha A/B region abolished ligand-induced beta-catenin degradation but not RXR alpha-mediated transactivation. RXR alpha-mediated inactivation of oncogenic beta-catenin paralleled a reduction in cell proliferation. These results suggest a potential role for RXR and its agonists in the regulation of beta-catenin turnover and related biological events.
Assuntos
Polipose Adenomatosa do Colo/metabolismo , Proteínas do Citoesqueleto/metabolismo , Receptores do Ácido Retinoico/metabolismo , Transativadores/metabolismo , Fatores de Transcrição/metabolismo , Western Blotting , Divisão Celular , Linhagem Celular , Reagentes de Ligações Cruzadas/farmacologia , Citosol/metabolismo , DNA Complementar/metabolismo , Relação Dose-Resposta a Droga , Deleção de Genes , Regulação da Expressão Gênica , Biblioteca Gênica , Genes Reporter , Quinase 3 da Glicogênio Sintase/metabolismo , Glicogênio Sintase Quinase 3 beta , Células HeLa , Humanos , Ligantes , Luciferases/metabolismo , Modelos Biológicos , Mutação , Plasmídeos/metabolismo , Ligação Proteica , Estrutura Terciária de Proteína , Receptores X de Retinoides , Fatores de Tempo , Transcrição Gênica , Ativação Transcricional , Transfecção , Proteína Supressora de Tumor p53/metabolismo , Técnicas do Sistema de Duplo-Híbrido , beta CateninaRESUMO
Agonists of retinoid X receptors (RXRs), which include the natural 9-cis-retinoic acid and synthetic analogs, are potent inducers of growth arrest and apoptosis in some cancer cells. As such, they are being used in clinical trials for the treatment and prevention of solid tumors and are used to treat cutaneous T cell lymphoma. However, the molecular mechanisms that underlie the anti-cancer effects of RXR agonists remain unclear. Here, we show that a novel pro-apoptotic pathway that is induced by RXR agonist is negatively regulated by casein kinase 1alpha (CK1alpha). CK1alpha associates with RXR in an agonist-dependent manner and phosphorylates RXR. The ability of an RXR agonist to recruit CK1alpha to a complex with RXR in cells correlates inversely with its ability to inhibit growth. Remarkably, depletion of CK1alpha in resistant cells renders them susceptible to RXR agonist-induced growth inhibition and apoptosis. Our study shows that CK1alpha can promote cell survival by interfering with RXR agonist-induced apoptosis. Inhibition of CK1alpha may enhance the anti-cancer effects of RXR agonists.