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Described is the development of a polymeric micelle drug delivery platform that addresses the physical property limitations of many nanovectors. The system employs triblock copolymers comprised of a hydrophilic poly(ethylene glycol) (PEG) block, and two poly(amino acid) (PAA) blocks: a stabilizing cross-linking central block, and a hydrophobic drug encapsulation block. Detailed description of synthetic strategies and considerations found to be critical are discussed. Of note, it was determined that the purity of the α-amino acid-N-carboxyanhydrides (NCA) monomers and PEG macroinitiator are ultimately responsible for impurities that arise during the polymerization. Also, contrary to current beliefs in the field, the presence of water does not adversely affect the polymerization of NCAs. Furthermore, we describe the impact of poly(amino acid) conformational changes, through the incorporation of d-amino acids to form mixed stereochemistry PAA blocks, with regard to the physical and pharmacokinetic properties of the resulting micelles.
Assuntos
Aminoácidos/química , Composição de Medicamentos/métodos , Sistemas de Liberação de Medicamentos/métodos , Compostos Férricos/química , Ácidos Hidroxâmicos/química , Polietilenoglicóis/química , Anidridos/química , Animais , Antineoplásicos/farmacocinética , Camptotecina/análogos & derivados , Camptotecina/farmacocinética , Daunorrubicina/farmacocinética , Liberação Controlada de Fármacos , Interações Hidrofóbicas e Hidrofílicas , Irinotecano , Micelas , Polimerização , Ratos , Ratos Endogâmicos F344 , EstereoisomerismoRESUMO
Nanoparticle drug carriers hold potential to improve current cancer therapy by delivering payload to the tumor environment and decreasing toxic side effects. Challenges in nanotechnology drug delivery include plasma instability, site-specific delivery, and relevant biomarkers. We have developed a triblock polymer comprising a hydroxamic acid functionalized center block that chelates iron to form a stabilized micelle that physically entraps chemotherapeutic drugs in the hydrophobic core. The iron-imparted stability significantly improves the integrity of the micelle and extends circulation pharmacokinetics in plasma over that of free drug. Furthermore, the paramagnetic properties of the iron-crosslinking exhibits contrast in the tumors for imaging by magnetic resonance. Three separate nanoparticle formulations demonstrate improved anti-tumor efficacy in xenograft models and decreased toxicity. We report a stabilized polymer micelle that improves the tolerability and efficacy of chemotherapeutic drugs, and holds potential for non-invasive MRI to image drug delivery and deposition in the tumor.
Assuntos
Antineoplásicos/farmacocinética , Neoplasias da Mama/diagnóstico por imagem , Neoplasias Colorretais/tratamento farmacológico , Portadores de Fármacos/química , Ferro/química , Micelas , Animais , Linhagem Celular Tumoral , Portadores de Fármacos/farmacocinética , Feminino , Humanos , Imageamento por Ressonância Magnética , Camundongos Nus , Polímeros/química , Ratos Sprague-Dawley , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Azido-functionalized poly(ethylene glycol) (PEG) derivatives are finding ever-increasing applications in the areas of conjugation chemistry and targeted drug delivery by their judicious incorporation into nanoparticle-forming polymeric systems. Quantification of azide incorporation into such PEG polymers is essential to their effective use. 1H Nuclear Magnetic Resonance (NMR) analysis offers the simplest approach; however, the relevant adjacent azide-bearing methylene protons are often obscured by the PEG manifold signals. This study describes the synthesis of 1,2,3-triazole adducts from their corresponding PEG azides via a convenient, mild click reaction, which facilitates straightforward NMR-based quantitative end-group analysis.This method was found to be compatible with many examples of bifunctional azido PEGs with molecular weights ranging from 2 to 18 kDa bearing a variety of functional groups. © 2016 The Authors. Journal of Polymer Science Part A: Polymer Chemistry Published by Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys. 2016, 54, 2888-2895.
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Epothilones have demonstrated promising potential for oncology applications but suffer from a narrow therapeutic window. Epothilone D stabilizes microtubules leading to apoptosis, is active against multidrug-resistant cells, and is efficacious in animal tumor models despite lack of stability in rodent plasma. Clinical development was terminated in phase II due to dose limiting toxicities near the efficacious dose. Taken together, this made epothilone D attractive for encapsulation in a stabilized polymer micelle for improved safety and efficacy. We have designed a library of triblock copolymers to develop IT-147, a lead formulation of epothilone D that extends plasma circulation for accumulation in the tumor environment, and potentially decrease systemic exposure to reduce dose limiting toxicities. The drug loading efficiency for IT-147 exceeds 90%, is 75 nm in diameter, and demonstrates pH-dependent release of epothilone D without chemical conjugation or enzymatic activation. Administration of IT-147 at 20 mg/kg increases exposure of epothilone D to the plasma compartment over 6-fold compared to free drug. At the same dose, 20 mg/kg epothilone D from IT-147 is considered the no observed adverse effect level (NOAEL) but is the maximum tolerated dose for free drug. Consequently, IT-147 is positioned to be a safer, more effective means to deliver epothilone D.
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Vaccination is regarded as one of the most cost-effective and reliable methods for combating disease. We have developed a new method for an oligomeric Aß-specific AD vaccination using polymer micelle-encapsulated peptide fragments, which overcome many problems of vaccination associated with the direct use of the Aß142 peptide. We studied different encapsulated forms of shortened Aß peptides with and without the entire T cell epitope in an APP/PS1 mouse model. After two inoculations with encapsulated Aß fragments, antibodies were produced in all mice with antibody titer greater than 1:12,800. No anti-polymer antibodies were detected after five inoculations, and none of the injected mice showed any adverse effects throughout experimentation. Anti-Aß antibodies from our polymer-encapsulated vaccine were able to bind to A plaques in the brain of our mice, and were able to specifically recognize oligomeric Aß. Our results suggest that the safety and efficacy issues previously encountered in other Aß vaccination trials may be successfully addressed by using micelle-encapsulated peptides. These shorter Aß fragments are also easier to synthesize and more cost-effective than the highly hydrophobic full-length Aß142 peptide.
Assuntos
Doença de Alzheimer/imunologia , Peptídeos beta-Amiloides/imunologia , Anticorpos/imunologia , Sistemas de Liberação de Medicamentos/métodos , Vacinas/imunologia , Doença de Alzheimer/prevenção & controle , Peptídeos beta-Amiloides/química , Peptídeos beta-Amiloides/metabolismo , Animais , Anticorpos/metabolismo , Encéfalo/imunologia , Modelos Animais de Doenças , Epitopos de Linfócito B , Camundongos , Camundongos Transgênicos , Micelas , Polímeros/química , Vacinas/química , Vacinas/metabolismoRESUMO
Recent emphasis has focused on the development of rationally designed polymer-based micelle carriers for drug delivery. The current work tests the hypothesis that target specificity can be enhanced by micelles with cancer-specific ligands. In particular, we describe the synthesis and characterization of a new gadolinium texaphyrin (Gd-Tx) complex encapsulated in an IVECT micellar system, stabilized through Fe(III) cross-linking and targeted with multiple copies of a specific ligand for the melanocortin 1 receptor (MC1R), which has been evaluated as a cell-surface marker for melanoma. On the basis of comparative MRI experiments, we have been able to demonstrate that these Gd-Tx micelles are able to target MC1R-expressing xenograft tumors in vitro and in vivo more effectively than various control systems, including untargeted or un-cross-linked Gd-Tx micelles. Taken in concert, the findings reported herein support the conclusion that appropriately designed micelles are able to deliver contrast agent payloads to tumors expressing the MC1R.
Assuntos
Imageamento por Ressonância Magnética/métodos , Micelas , Polímeros/química , Receptor Tipo 1 de Melanocortina/química , Animais , Linhagem Celular Tumoral , Humanos , Camundongos , Camundongos SCIDRESUMO
Chemotherapeutic drugs are widely used for the treatment of cancer; however, use of these drugs is often associated with patient toxicity and poor tumor delivery. Micellar drug carriers offer a promising approach for formulating and achieving improved delivery of hydrophobic chemotherapeutic drugs; however, conventional micelles do not have long-term stability in complex biological environments such as plasma. To address this problem, a novel triblock copolymer has been developed to encapsulate several different hydrophobic drugs into stable polymer micelles. These micelles have been engineered to be stable at low concentrations even in complex biological fluids, and to release cargo in response to low pH environments, such as in the tumor microenvironment or in tumor cell endosomes. The particle sizes of drugs encapsulated ranged between 30-80 nm, with no relationship to the hydrophobicity of the drug. Stabilization of the micelles below the critical micelle concentration was demonstrated using a pH-reversible crosslinking mechanism, with proof-of-concept demonstrated in both in vitro and in vivo models. Described herein is polymer micelle drug delivery system that enables encapsulation and stabilization of a wide variety of chemotherapeutic drugs in a single platform.
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Polymer micelles are promising drug delivery vehicles for the delivery of anticancer agents to tumors. Often, anticancer drugs display potent cytotoxic effects towards cancer cells but are too hydrophobic to be administered in the clinic as a free drug. To address this problem, a polymer micelle was designed using a triblock copolymer (ITP-101) that enables hydrophobic drugs to be encapsulated. An SN-38 encapsulated micelle, IT-141, was prepared that exhibited potent in vitro cytotoxicity against a wide array of cancer cell lines. In a mouse model, pharmacokinetic analysis revealed that IT-141 had a much longer circulation time, plasma exposure, and tumor exposure compared to irinotecan. IT-141 was also superior to irinotecan in terms of antitumor activity, exhibiting greater tumor inhibition in HT-29 and HCT116 colorectal cancer xenograft models at half the dose of irinotecan. The antitumor effect of IT-141 was dose-dependent and caused complete growth inhibition and tumor regression at well-tolerated doses. Varying the specific concentration of SN-38 within the IT-141 micelle had no detectible effect on this antitumor activity, indicating no differences in activity between different IT-141 formulations. In summary, IT-141 is a potent micelle-based chemotherapy that holds promise for the treatment of colorectal cancer.
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The incidence of malignant melanoma is rising faster than that of any other cancer in the United States. Because of its high expression on the surface of melanomas, MC1R has been investigated as a target for selective imaging and therapeutic agents against melanoma. Eight ligands were screened against cell lines engineered to overexpress MC1R, MC4R, or MC5R. Of these, compound 1 (4-phenylbutyryl-His-dPhe-Arg-Trp-NH(2)) exhibited high (0.2 nM) binding affinity for MC1R and low (high nanomolar) affinities for MC4R and MC5R. Functionalization of the ligand at the C-terminus with an alkyne for use in Cu-catalyzed click chemistry was shown not to affect the binding affinity. Finally, formation of the targeted polymer, as well as the targeted micelle formulation, also resulted in constructs with low nanomolar binding affinity.
Assuntos
Antineoplásicos/síntese química , Azidas/química , Oligopeptídeos/síntese química , Polietilenoglicóis/química , Proteínas/química , Receptor Tipo 1 de Melanocortina/metabolismo , Alcinos/química , Antineoplásicos/química , Antineoplásicos/farmacologia , Ligação Competitiva , Química Click , Ensaios de Seleção de Medicamentos Antitumorais , Células HCT116 , Células HEK293 , Humanos , Ligantes , Melanoma , Micelas , Oligopeptídeos/química , Oligopeptídeos/farmacologia , Receptor Tipo 4 de Melanocortina/metabolismo , Receptores de Melanocortina/metabolismo , Relação Estrutura-AtividadeRESUMO
Size-correlated single-molecule fluorescence measurements on CdSe quantum dots functionalized with oligo(phenylene vinylene) (OPV) ligands exhibit modified fluorescence intermittency (blinking) statistics that are highly sensitive to the degree of ligand coverage on the quantum dot surface. As evidenced by a distinct surface height signature, fully covered CdSe-OPV nanostructures (approximately 25 ligands) show complete suppression of blinking in the solid state on an integration time scale of 1 s. Some access to dark states is observed on finer time scales (100 ms) with average persistence times significantly shorter than those from ZnS-capped CdSe quantum dots. This effect is interpreted as resulting from charge transport from photoexcited OPV into vacant trap sites on the quantum dot surface. These results suggest exciting new applications of composite quantum dot/organic systems in optoelectronic systems.
Assuntos
Compostos de Cádmio/química , Nanoestruturas/química , Polivinil/química , Pontos Quânticos , Compostos de Selênio/química , Fluorescência , Ligantes , Tamanho da Partícula , Espectrometria de Fluorescência , Fatores de TempoRESUMO
The temporal and spectral properties of luminescence from individual CdSe quantum dot-oligophenylene vinylene nanostructures (single quantum dots with conjugated organic ligands coordinated to the surface) are profoundly modified relative to blended films of the same components. These kinds of composite quantum dot-conjugated organic systems have attracted significant interest as a way to improve efficiency in photovoltaic device applications. By direct functionalization of the dot surface with the conjugated organic ligands, we realize a significant enhancement in energy transfer and luminescence stability.
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The organization of inorganic nanostructures within self-assembled organic or biological templates is receiving the attention of scientists interested in developing functional hybrid materials. Previous efforts have concentrated on using such scaffolds to spatially arrange nanoscopic elements as a strategy for tailoring the electrical, magnetic or photonic properties of the material. Recent theoretical arguments have suggested that synergistic interactions between self-organizing particles and a self-assembling matrix material can lead to hierarchically ordered structures. Here we show that mixtures of diblock copolymers and either cadmium selenide- or ferritin-based nanoparticles exhibit cooperative, coupled self-assembly on the nanoscale. In thin films, the copolymers assemble into cylindrical domains, which dictate the spatial distribution of the nanoparticles; segregation of the particles to the interfaces mediates interfacial interactions and orients the copolymer domains normal to the surface, even when one of the blocks is strongly attracted to the substrate. Organization of both the polymeric and particulate entities is thus achieved without the use of external fields, opening a simple and general route for fabrication of nanostructured materials with hierarchical order.
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In polymernanoparticle composites, uniform dispersion of the nanoparticles carries advantages over cases where nanoparticle aggregation dominates. Such dispersion has been particularly difficult to obtain in the case of composites prepared from nanoparticles and conjugated polymers. Here, we show that cadmium selenide nanocrystals, or quantum dots, can be integrated into thin films of poly(para-phenylene vinylene) (PPV) without aggregation. The two key departures from previous studies of quantum-dot/electronic polymer composites are (1) the synthesis of high-quality quantum dots directly in novel, functional ligands, thus eliminating the need for ligand exchange, and (2) polymerization chemistry that grafts PPV to the quantum dot surface. Solid-state photoluminescence spectra of composite materials prepared by these novel techniques reveal the critical importance of the quantum dot-polymer interface that will enable new investigations in nanoparticle-based light-emitting devices.