RESUMO
Tenofovir (TFV) is an adenosine nucleotide analog with activity against HIV and HSV-2. Secondary analyses of clinical trials evaluating TFV gel as pre-exposure prophylaxis (PrEP) for HIV have shown that gel formulations of TFV provide significant protection against both HIV and HSV-2 acquisition in women who had evidence of use. An alternate quick-dissolving polymeric thin film, to deliver TFV (20 and 40â mg) has been developed as a potential multipurpose technology (MPT) platform. Film formulation was developed based on excipient compatibility, stability, and ability to incorporate TFV doses. Placebo, low dose (20â mg), and high dose (40â mg) films were utilized in these studies. The developed film platform efficiently incorporated the high dose of TFV (40â mg/film), released more than 50% of drug in 15â min with no in vitro toxicity. Pharmacological activity was confirmed in an ex vivo HIV-1 challenge study, which showed a reduction in HIV-1 infection with TFV films. Films were stable at both doses for at least 2 years. These films were found to be safe in macaques with repeated exposure for 2 weeks as evidenced by minimal perturbation to tissues, microbiome, neutrophil influx, and pH. Macaque sized TFV film (11.2â mg) evaluated in a pigtail macaque model showed higher vaginal tissue concentrations of TFV and active TFV diphosphate compared to a 15â mg TFV loaded gel. These studies confirm that TFV films are stable, safe and efficiently deliver the drug in cervicovaginal compartments supporting their further clinical development.
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The female upper genital tract (UGT) hosts important reproductive organs including the cervix, uterus, fallopian tubes, and ovaries. Several pathologies affect these organ systems such as infections, reproductive issues, structural abnormalities, cancer, and inflammatory diseases that could have significant impact on women's overall health. Effective disease management is constrained by the multifaceted nature of the UGT, complex anatomy and a dynamic physiological environment. Development of drug delivery strategies that can overcome mucosal and safety barriers are needed for effective disease management. This review introduces the anatomy, physiology, and mucosal properties of the UGT and describes drug delivery barriers, advances in drug delivery technologies, and opportunities available for new technologies that target the UGT.
Assuntos
Sistemas de Liberação de Medicamentos , Doenças dos Genitais Femininos/tratamento farmacológico , Animais , Feminino , Genitália Feminina/anatomia & histologia , Genitália Feminina/metabolismo , Humanos , Mucosa/metabolismo , Saúde da MulherRESUMO
Optical imaging can be utilized for several pharmaceutical applications involving near-infrared fluorescent (NIRF) dyes or NIRF moiety-containing products. Especially during the early phases of product development, NIRF dyes can be used as surrogates for drugs and optical imaging methods can be utilized to optimize the pharmaceutical product properties based on dye entrapment efficiency, in vitro dye release, cellular uptake, and in vivo biodistribution. Based on in vivo accumulation, product efficacy and toxicity can be evaluated in the early development stage. Compared to visible fluorescent dyes, NIRF offers advantages such as low background from formulation excipients as well as biological components.In this chapter, the utility of NIRF imaging methods for in vitro characterization (in vitro release and cellular uptake) and in vivo/ex vivo applicability of pharmaceutically relevant products is presented in detail. Specifically, the application of fluorescence imaging to characterize perfluorocarbon-based formulations for dye loading, in vitro release, cellular uptake, and in vivo imaging to assess target accumulation and biodistribution is discussed. These methods are widely applicable to other nanoparticle-based products involved in inflammation/cancer imaging and therapy. Overall, NIRF-based techniques are indispensible because they are relatively easy, fast, and cost effective to characterize and optimize pharmaceutical products at different stages of product development.
Assuntos
Probióticos/farmacocinética , Tomografia Óptica/métodos , Animais , Corantes Fluorescentes/química , Fluorocarbonos/química , Humanos , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Hidrogel de Polietilenoglicol-Dimetacrilato/farmacocinética , Camundongos , Probióticos/química , Distribuição TecidualRESUMO
Targeting macrophages for therapeutic and diagnostic purposes is an attractive approach applicable to multiple diseases. Here, we present a theranostic nanoemulsion platform for simultaneous delivery of an anti-inflammatory drug (celecoxib) to macrophages and monitoring of macrophage migration patterns by optical imaging, as measurement of changes in inflammation. The anti-inflammatory effect of the theranostic nanoemulsions was evaluated in a mouse inflammation model induced with complete Freund's adjuvant (CFA). Nanoemulsions showed greater accumulation in the inflamed vs. control paw, with histology confirming their specific localization in CD68 positive macrophages expressing cyclooxygenase-2 (COX-2) compared to neutrophils. With a single dose administration of the celecoxib-loaded theranostic, we observed a reduction in fluorescence in the paw with time, corresponding to a reduction in macrophage infiltration. Our data strongly suggest that delivery of select agents to infiltrating macrophages can potentially lead to new treatments of inflammatory diseases where macrophage behavior changes are monitored in vivo.
Assuntos
Celecoxib/administração & dosagem , Inibidores de Ciclo-Oxigenase 2/administração & dosagem , Portadores de Fármacos , Inflamação/tratamento farmacológico , Macrófagos/imunologia , Nanotecnologia/métodos , Animais , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Movimento Celular/imunologia , Ciclo-Oxigenase 2/metabolismo , Modelos Animais de Doenças , Emulsões , Feminino , Ativação de Macrófagos/efeitos dos fármacos , Ativação de Macrófagos/imunologia , Camundongos , Neutrófilos/imunologiaRESUMO
Inflammatory disease management poses challenges due to the complexity of inflammation and inherent patient variability, thereby necessitating patient-specific therapeutic interventions. Theranostics, which integrate therapeutic and imaging functionalities, can be used for simultaneous imaging and treatment of inflammatory diseases. Theranostics could facilitate assessment of safety, toxicity and real-time therapeutic efficacy leading to personalized treatment strategies. Macrophages are an important cellular component of inflammatory diseases, participating in varied roles of disease exacerbation and resolution. The inherent phagocytic nature, abundance and disease homing properties of macrophages can be targeted for imaging and therapeutic purposes. This review discusses the utility of theranostics in macrophage ablation, phenotype modulation and inhibition of their inflammatory activity leading to resolution of inflammation in several diseases.
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Diagnóstico por Imagem/métodos , Sistemas de Liberação de Medicamentos , Inflamação/diagnóstico , Inflamação/tratamento farmacológico , Macrófagos/imunologia , Nanomedicina/métodos , Medicina de Precisão/métodos , HumanosRESUMO
Chronic neuropathic pain following surgery represents a serious worldwide health problem leading to life-long treatment and the possibility of significant disability. In this study, neuropathic pain was modeled using the chronic constriction injury (CCI). The CCI rats exhibit mechanical hypersensitivity (typical neuropathic pain symptom) to mechanical stimulation of the affected paw 11 days post surgery, at a time when sham surgery animals do not exhibit hypersensitivity. Following a similar time course, TRPV1 gene expression appears to rise with the hypersensitivity to mechanical stimulation. Recent studies have shown that immune cells play a role in the development of neuropathic pain. To further explore the relationship between neuropathic pain and immune cells, we hypothesize that the infiltration of immune cells into the affected sciatic nerve can be monitored in vivo by molecular imaging. To test this hypothesis, an intravenous injection of a novel perfluorocarbon (PFC) nanoemulsion, which is phagocytosed by inflammatory cells (e.g. monocytes and macrophages), was used in a rat CCI model. The nanoemulsion carries two distinct imaging agents, a near-infrared (NIR) lipophilic fluorescence reporter (DiR) and a ¹9F MRI (magnetic resonance imaging) tracer, PFC. We demonstrate that in live rats, NIR fluorescence is concentrated in the area of the affected sciatic nerve. Furthermore, the ¹9FF MRI signal was observed on the sciatic nerve. Histological examination of the CCI sciatic nerve reveals significant infiltration of CD68 positive macrophages. These results demonstrate that the infiltration of immune cells into the sciatic nerve can be visualized in live animals using these methods.
Assuntos
Diagnóstico por Imagem/métodos , Imageamento por Ressonância Magnética/métodos , Neuralgia/patologia , Neuropatia Ciática/diagnóstico , Animais , Modelos Animais de Doenças , Emulsões , Fluorescência , Flúor , Fluorocarbonos/administração & dosagem , Humanos , Raios Infravermelhos , Macrófagos/metabolismo , Espectroscopia de Ressonância Magnética/métodos , Masculino , Microscopia Confocal , Monócitos/metabolismo , Nanopartículas/administração & dosagem , Neuralgia/metabolismo , Neuralgia/fisiopatologia , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Nervo Isquiático/lesões , Nervo Isquiático/metabolismo , Nervo Isquiático/fisiopatologia , Neuropatia Ciática/metabolismo , Neuropatia Ciática/fisiopatologia , Sensibilidade e EspecificidadeRESUMO
Cylcooxgenase-2 (COX-2) expressing macrophages, constituting a major portion of tumor mass, are involved in several pro-tumorigenic mechanisms. In addition, macrophages are actively recruited by the tumor and represent a viable target for anticancer therapy. COX-2 specific inhibitor, celecoxib, apart from its anticancer properties was shown to switch macrophage phenotype from tumor promoting to tumor suppressing. Celecoxib has low aqueous solubility, which may limit its tumor inhibiting effect. As opposed to oral administration, we propose that maximum anticancer effect may be achieved by nanoemulsion mediated intravenous delivery. Here we report multifunctional celecoxib nanoemulsions that can be imaged by both near-infrared fluorescence (NIRF) and (19)F magnetic resonance. Celecoxib loaded nanoemulsions showed a dose dependent uptake in mouse macrophages as measured by (19)F NMR and NIRF signal intensities of labeled cells. Dramatic inhibition of intracellular COX-2 enzyme was observed in activated macrophages upon nanoemulsion uptake. COX-2 enzyme inhibition was statistically equivalent between free drug and drug loaded nanoemulsion. However, nanoemulsion mediated drug delivery may be advantageous, helping to avoid systemic exposure to celecoxib and related side effects. Dual molecular imaging signatures of the presented nanoemulsions allow for future in vivo monitoring of the labeled macrophages and may help in examining the role of macrophage COX-2 inhibition in inflammation-cancer interactions. These features strongly support the future use of the presented nanoemulsions as anti-COX-2 theranostic nanomedicine with possible anticancer applications.
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Inibidores de Ciclo-Oxigenase 2/química , Éteres/química , Etilenoglicol/química , Fluorocarbonos/química , Nanomedicina , Animais , Celecoxib , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Inibidores de Ciclo-Oxigenase 2/administração & dosagem , Inibidores de Ciclo-Oxigenase 2/toxicidade , Sistemas de Liberação de Medicamentos , Emulsões , Macrófagos/efeitos dos fármacos , Camundongos , Ressonância Magnética Nuclear Biomolecular , Tamanho da Partícula , Pirazóis/administração & dosagem , Pirazóis/química , Sulfonamidas/administração & dosagem , Sulfonamidas/químicaRESUMO
Haloenol pyran-2-ones and morpholin-2-ones were synthesized and evaluated as inhibitors of cell growth in two different prostate human cancer cell lines (PC-3 and LNCaP). Analogs derived from L- and D-phenylglycine were found to be the most effective antagonists of LNCaP and PC-3 cell growth. Additional studies reveal that the inhibitors induced G2/M arrest and the (S)-enantiomer of the phenylglycine-based derivatives was a more potent inhibitor of cytosolic iPLA(2)ß.