RESUMO
Achieving the best possible outcome for the therapy is the main goal of a medicine. Therefore, nanocarriers and co-delivery strategies were invented to meet this need, as they can benefit many diseases. This approach was applied specifically for cancer treatment, with some success. However, these strategies may benefit many other clinical issues. Skin is the largest and most exposed organ of the human body, with physiological and psychological properties. Due to its exposition and importance, it is not difficult to understand how many skin diseases may impact on patients' lives, representing an important burden for society. Thus, this review aims to summarize the state of the art in research concerning nanocarriers and co-delivery strategies for topical agents' applications targeting skin diseases. The challenge for the medicine of the future is to deliver the drug with spatial and temporal control. Therefore, the co-encapsulation of drugs and the appropriate form of administration for them are so important and remain as unmet needs.
Assuntos
Nanopartículas , Dermatopatias , Humanos , Preparações Farmacêuticas/metabolismo , Pele/metabolismo , Absorção Cutânea , Dermatopatias/metabolismo , Sistemas de Liberação de Medicamentos , Portadores de Fármacos/metabolismo , Administração Cutânea , Administração TópicaRESUMO
Cancer metastasis and secondary tumor initiation largely depend on circulating tumor cell (CTC) and vascular endothelial cell (EC) interactions by incompletely understood mechanisms. Endothelial glycocalyx (GCX) dysfunction may play a significant role in this process. GCX structure depends on vascular flow patterns, which are irregular in tumor environments. This work presents evidence that disturbed flow (DF) induces GCX degradation, leading to CTC homing to the endothelium, a first step in secondary tumor formation. A 2-fold greater attachment of CTCs to human ECs was found to occur under DF conditions, compared to uniform flow (UF) conditions. These results corresponded to an approximately 50% decrease in wheat germ agglutinin (WGA)-labeled components of the GCX under DF conditions, vs UF conditions, with undifferentiated levels of CTC-recruiting E-selectin under DF vs UF conditions. Confirming the role of the GCX, neuraminidase induced the degradation of WGA-labeled GCX under UF cell culture conditions or in Balb/C mice and led to an over 2-fold increase in CTC attachment to ECs or Balb/C mouse lungs, respectively, compared to untreated conditions. These experiments confirm that flow-induced GCX degradation can enable metastatic CTC arrest. This work, therefore, provides new insight into pathways of secondary tumor formation.
Assuntos
Neoplasias da Mama/patologia , Endotélio Vascular/patologia , Glicocálix/metabolismo , Hemodinâmica , Neoplasias Pulmonares/secundário , Células Neoplásicas Circulantes/patologia , Neuraminidase/metabolismo , Animais , Neoplasias da Mama/metabolismo , Células Cultivadas , Selectina E/metabolismo , Endotélio Vascular/metabolismo , Feminino , Humanos , Neoplasias Pulmonares/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Células Neoplásicas Circulantes/metabolismoRESUMO
Dendritic cells serve as the main immune cells that trigger the immune response. We developed a simple and cost-effective nanovaccine platform based on the α1',2-mannobiose derivative for dendritic cell targeting. In previous work, we have formulated the α1,2-mannobiose-based nanovaccine platform with plasmid DNA and tested it in cattle against BoHV-1 infection. There, we have shown that the dendritic cell targeting using this nanovaccine platform in vivo can boost the immunogenicity, resulting in a long-lasting immunity. In this work, we aim to characterize the α1',2-mannobiose derivative, which is key in the nanovaccine platform. This DC-targeting strategy takes advantage of the specific receptor known as DC-SIGN and exploits its capacity to bind α1,2-mannobiose that is present at terminal ends of oligosaccharides in certain viruses, bacteria, and other pathogens. The oxidative conjugation of α1',2-mannobiose to NH2-PEG2kDa-DSPE allowed us to preserve the chemical structure of the non-reducing mannose of the disaccharide and the OH groups and the stereochemistry of all carbons of the reducing mannose involved in the binding to DC-SIGN. Here, we show specific targeting to DC-SIGN of decorated micelles incubated with the Raji/DC-SIGN cell line and uptake of targeted liposomes that took place in human, bovine, mouse, and teleost fish DCs in vitro, by flow cytometry. Specific targeting was found in all cultures, demonstrating a species-non-specific avidity for this ligand, which opens up the possibility of using this nanoplatform to develop new vaccines for various species, including humans.
Assuntos
Células Apresentadoras de Antígenos/imunologia , Moléculas de Adesão Celular/imunologia , Células Dendríticas/imunologia , Lectinas Tipo C/imunologia , Linfoma/imunologia , Manose/química , Receptores de Superfície Celular/imunologia , Vacinas/imunologia , Animais , Bovinos , Feminino , Peixes , Humanos , Linfoma/terapia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Especificidade da Espécie , Vacinas/administração & dosagemRESUMO
PURPOSE: To develop immunoliposomes modified with monoclonal cancer-specific antibody (mAb) 2C5 and co-loaded with a combination of two chemotherapeutics, in order to simultaneously target bulk cancer cells using paclitaxel and cancer stem cells (CSCs) using salinomycin to prevent cancer growth and metastases. METHODS: Breast cancer cells (MDA-MB-231 and/or SK-BR-3) were chosen as models for all in vitro testing. Liposomes composed of natural phospholipids co-loaded with salinomycin and paclitaxel were prepared and physically characterized. Immunoliposomes modified with mAb 2C5 coupled to polymeric conjugate were prepared and characterized for specific targeting. Wound healing assay was performed using the combination of free drugs in vitro. In vitro studies on cellular interaction and uptake were followed by holographic imaging to study cell-killing, cell-division and proliferation inhibiting effects of the formulation. Ex-vivo study on hemolysis was investigated to check possible toxicity of the formulation. RESULTS: Physical characterization of the liposomes showed stable nanoparticles of consistent and desirable size range (170-220 nm), zeta potential (-13 mV to - 20 mV), polydispersity indices (<0.2) and drug encapsulation efficiencies (~150 µg per ml for salinomycin, ~210 µg/ml for paclitaxel and 1:1 for combination drug loaded liposomes). Combination therapy strongly affected cancer cell proliferation as shown by significant diminishing of artificial gap closure at the wound site on MDA-MB-231 cells in culture using wound healing assay. Quantitation of changes in wound widths showed ~219 µm for drug combination, ~104 µm for only paclitaxel, and ~ 7 µm for only salinomycin treatments. Statistically significant increase in cellular interaction and specific uptake of the targeted drug co-loaded liposomal nanopreparation (p value ≤ 0.05) by MDA-MB-231 and SK-BR-3 cells confirmed the effectiveness of the approach. Holographic imaging using MDA-MB-231 cells produced visible increase in cell-killing, proliferation and division in vitro. Ex-vivo experimentation showed reduced hemolysis correlating with low toxicity in athymic nude mice model. CONCLUSION: The results demonstrated the enhanced therapeutic efficacy of a combination of salinomycin and paclitaxel delivered by mAb 2C5-modified liposomal preparation in cancer therapy.
Assuntos
Anticorpos Monoclonais/química , Protocolos de Quimioterapia Combinada Antineoplásica/química , Portadores de Fármacos/química , Lipossomos/química , Paclitaxel/química , Piranos/química , Animais , Anticorpos Monoclonais/imunologia , Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Permeabilidade da Membrana Celular , Proliferação de Células/efeitos dos fármacos , Preparações de Ação Retardada/química , Composição de Medicamentos , Liberação Controlada de Fármacos , Feminino , Humanos , Lipossomos/imunologia , Camundongos Nus , Neoplasias Experimentais , Paclitaxel/farmacologia , Fosfatidilcolinas/química , Polietilenoglicóis/química , Piranos/farmacologiaRESUMO
Targeted delivery of chemotherapeutics to tumors has the potential to reach a high dose at the tumor while minimizing systemic exposure. Incorporation of antibody within a micellar platform represents a drug delivery system for tumor-targeted delivery of antitumor agents. Such modified immunomicelles can result in an increased accumulation of antitumor agents and enhanced cytotoxicity toward cancer cells. Here, mixed dendrimer micelles (MDM) composed of PEG2k-DOPE-conjugated generation 4 polyamidoamine dendrimer G4-PAMAM-PEG2k-DOPE and PEG5k-DOPE were coloaded with doxorubicin and siMDR-1. This formulation was further modified with monoclonal antibodies 2C5 with nucleosome-restricted specificity that effectively recognized cancer cells via the cell-surface-bound nucleosomes. Micelles with attached 2C5 antibodies significantly enhanced cellular association and tumor killing in both monolayer and spheroid tumor models as well as in vivo in experimental animals compared to the nontargeted formulations.
Assuntos
Anticorpos Monoclonais/química , Antineoplásicos/administração & dosagem , Dendrímeros/química , Sistemas de Liberação de Medicamentos , Micelas , Neoplasias Experimentais/tratamento farmacológico , RNA Interferente Pequeno/administração & dosagem , Animais , Anticorpos Monoclonais/administração & dosagem , Anticorpos Monoclonais/farmacocinética , Linhagem Celular Tumoral , Doxorrubicina/administração & dosagem , Composição de Medicamentos , Feminino , Humanos , Camundongos , Neoplasias Experimentais/patologia , Esferoides Celulares , Distribuição TecidualRESUMO
PURPOSE: Since the last decade, it is established that nonspecific delivery of chemotherapeutics fails to effectively treat cancer due to systemic cytotoxicity, poor biodistribution at tumor site and most importantly the development of drug resistance (MDR). Stimuli-sensitive drug delivery systems gained significant attention in recent years for effective tumor therapy and reversal of MDR. The aim of this study was developing a redox sensitive micellar prodrug system, by taking the advantage of the significant difference in GSH levels between extracellular and intracellular environments, but more importantly in healthy and tumor tissues. METHODS: Redox sensitive PEG2000-S-S-PTX micelles were developed for intracellular paclitaxel delivery and characterized in vitro. In vitro release studies were carried out and followed by cytotoxicity studies in chemo-resistant ovarian and breast cancer cells in various reducing environments for different time periods to confirm their potential. RESULTS: PEG2000-S-S-PTX, was synthesized and characterized as a redox sensitive micellar prodrug system. The reduction sensitivity and in vitro PTX release properties were confirmed in reducing environments comparatively with physiological conditions. Cytotoxicity studies suggested that ovarian (SK-OV-3) cells could be better candidates for treatment with redox-sensitive drug delivery systems than breast (MCF-7) cancer cells. CONCLUSIONS: The results of this study highlights the importance of personalized therapy since no fits-for-all system can be developed for different cancer with significantly different metabolic activities. Graphical Abstract Schematic representation of self-assembly of reduction-sensitive PEG2000-S-S-PTX micelles and GSH dependent release of PTX.
Assuntos
Antineoplásicos Fitogênicos/administração & dosagem , Neoplasias da Mama/tratamento farmacológico , Preparações de Ação Retardada/química , Dissulfetos/química , Neoplasias Ovarianas/tratamento farmacológico , Paclitaxel/administração & dosagem , Antineoplásicos Fitogênicos/farmacologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Resistência a Múltiplos Medicamentos , Resistencia a Medicamentos Antineoplásicos , Feminino , Humanos , Micelas , Oxirredução , Paclitaxel/farmacologia , Polietilenoglicóis/químicaRESUMO
Folic acid is often used for active targeting of tumor cells to enhance therapeutic outcomes. Here, folic acid was conjugated with chitosan and folate-conjugated chitosan-lipid hybrid nanoparticles were prepared by ionic gelation method using anionic lipid. These nanoparticles were in size range of 200 to 400 nm with spherical shape. In vitro drug release data suggested a sustained release of cisplatin. The therapeutic efficacy of the folate-conjugated hybrid nanoparticles was evaluated in SK-OV-3, A2780 and MCF-7 cancer cell lines. A significant increase in cytotoxicity was observed with folate targeted LPHNPs compared to non-targeted LPHNPs. Significantly enhanced cellular uptake and cell cycle arrest resulting from folate-targeted nanoparticles were confirmed using fluorescence microscopy and flow cytometry. The therapeutic efficacy and tumor penetration were further evaluated in 3D spheroid tumor models. These studies suggest that folate-conjugated lipid-chitosan nanoparticles could enhance therapeutic activity and may represent a promising platform for active targeting of tumor cells.
Assuntos
Quitosana/química , Cisplatino/química , Ácido Fólico/química , Nanopartículas/química , Polímeros/química , Linhagem Celular Tumoral , Sistemas de Liberação de Medicamentos/métodos , Citometria de Fluxo , Humanos , Células MCF-7 , Microscopia de Fluorescência , Esferoides Celulares/efeitos dos fármacosRESUMO
The development of polymer-based nanoparticulate delivery systems for siRNA is important for the clinical success of gene therapy. However, there are some major drawbacks that need to be overcome. Short interfering RNA (siRNA) has been investigated as a potential therapeutic drug to silence disease-associated genes, but its usage is limited due to the lack of effective and safe nanocarriers. In this study, DOPE-PEI, a nanoparticle consisting of the fusogenic lipid 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) conjugated with low-molecular-weight, 600 Da, branched polyethylenimine (PEI) was produced and optimized for siRNA delivery. This delivery system was modified with other components such as 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine-N-[methoxy(polyethyleneglycol)2000] (DOPE-PEG2K), DOPE-PEG3.4K-bombesin and 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine/1,2-dioleoyl-3-trimethylammonium-propane (DOPE/DOTAP) and tested on PC-3 cells. The conjugation of DOPE to PEI polymer (DOPE-PEI) improved the efficiency of PEI to deliver siRNA into the cytosol and knockdown genes, but demonstrated high toxicity. The addition of DOPE-PEG2K reduced cellular toxicity by masking the surface positive charge of the DOPE-PEI/siRNA complex, with the incorporation of a gastrin-releasing peptide receptor (GRPR) targeting peptide and DOPE/DOTAP components improving the cellular uptake of siRNA into targeted cells and the siRNA knockdown efficiency.
Assuntos
Nanopartículas/química , Peptídeos/química , Polímeros/química , RNA Interferente Pequeno/administração & dosagem , Linhagem Celular Tumoral , Portadores de Fármacos/química , Ácidos Graxos Monoinsaturados/química , Técnicas de Silenciamento de Genes , Técnicas de Transferência de Genes , Terapia Genética/métodos , Humanos , Iminas/química , Lipídeos/química , Células PC-3 , Fosfatidiletanolaminas/química , Polietilenoglicóis/química , Polietilenos/química , Compostos de Amônio Quaternário/química , Receptores da Bombesina/metabolismoRESUMO
Conventional drug delivery approaches are plagued by issues pertaining to systemic toxicity and repeated dosing. Hydrogels offer convenient drug delivery vehicles to ensure these disadvantages are minimized and the therapeutic benefits from the drug are optimized. With exquisitely tunable physical properties that confer them great controlled drug release features and the merits they offer for labile drug protection from degradation, hydrogels emerge as very efficient drug delivery systems. The versatility and diversity of the hydrogels extend their applications beyond targeted drug delivery also to wound dressings, contact lenses and tissue engineering to name but a few. They are 90% water, and highly porous to accommodate drugs for delivery and facilitate controlled release. Herein we discuss hydrogels and how they could be manipulated for targeted drug delivery applications. Suitable examples from the literature are provided that support the recent advancements of hydrogels in targeted drug delivery in diverse disease areas and how they could be suitably modified in very different ways for achieving significant impact in targeted drug delivery. With their enormous amenability to modification, hydrogels serve as promising delivery vehicles of therapeutic molecules in several disease conditions, including cancer and diabetes.
Assuntos
Portadores de Fármacos , Sistemas de Liberação de Medicamentos , Hidrogéis , Animais , Portadores de Fármacos/química , Liberação Controlada de Fármacos , Humanos , Hidrogéis/química , Interações Hidrofóbicas e Hidrofílicas , Espécies Reativas de Oxigênio/químicaRESUMO
Multiple factors are involved in the development of cancers and their effects on survival rate. Many are related to chemo-resistance of tumor cells. Thus, treatment with a single therapeutic agent is often inadequate for successful cancer therapy. Ideally, combination therapy inhibits tumor growth through multiple pathways by enhancing the performance of each individual therapy, often resulting in a synergistic effect. Polymeric nanoparticles prepared from block co-polymers have been a popular platform for co-delivery of combinations of drugs associated with the multiple functional compartments within such nanoparticles. Various polymeric nanoparticles have been applied to achieve enhanced therapeutic efficacy in cancer therapy. However, reported drug ratios used in such systems often vary widely. Thus, the same combination of drugs may result in very different therapeutic outcomes. In this review, we investigated polymeric co-delivery systems used in cancer treatment and the drug combinations used in these systems for synergistic anti-cancer effect. Development of polymeric co-delivery systems for a maximized therapeutic effect requires a deeper understanding of the optimal ratio among therapeutic agents and the natural heterogenicity of tumors.
Assuntos
Antineoplásicos/uso terapêutico , Nanopartículas/química , Neoplasias/tratamento farmacológico , Animais , Antineoplásicos/química , Antineoplásicos/farmacologia , Sistemas de Liberação de Medicamentos/métodos , Cálculos da Dosagem de Medicamento , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Sinergismo Farmacológico , Humanos , Tamanho da Partícula , Polímeros/química , Polímeros/farmacologia , Polímeros/uso terapêutico , Transdução de Sinais/efeitos dos fármacosRESUMO
Gene therapy has emerged as an alternative in the treatment of cancer, particularly in cases of resistance to chemo and radiotherapy. Different approaches to deliver genetic material to tumor tissues have been proposed, including the use of small non-coding RNAs due to their multiple mechanisms of action. However, such promise has shown limits in in vivo application related to RNA's biological instability and stimulation of immunity, urging the development of systems able to overcome those barriers. In this review, we discuss the use of RNA interference in cancer therapy with special attention to the role of siRNA and miRNA and to the challenges of their delivery in vivo. We introduce a promising class of drug delivery system known as micelle-like nanoparticles and explore their synthesis and advantages for gene therapy as well as the recent findings in in vitro, in vivo and clinical studies.
Assuntos
Sistemas de Liberação de Medicamentos , Micelas , MicroRNAs/metabolismo , Nanopartículas/química , Neoplasias/terapia , RNA Interferente Pequeno/metabolismo , Animais , Linhagem Celular Tumoral , Quitosana/química , Ensaios Clínicos Fase I como Assunto , Ensaios Clínicos Fase II como Assunto , Modelos Animais de Doenças , Humanos , Poliaminas/química , Polietilenoimina/química , Polilisina/químicaRESUMO
A label-free, high content, time-lapse holographic imaging system was applied to studies in pharmaceutical compound development. Multiple fields of cellular images are obtained over typically several day evaluations within standard CO2 incubators. Events are segmented to obtain population data of cellular features, which are displayed in scattergrams and histograms. Cell tracking is accomplished, accompanied by Cartesian plots of cell movement, as well as plots of cell features vs. time in novel 4-D displays of X position, Y position, time, and cell thickness. Our review of the instrument validation data includes 1) tracking of Giant HeLa cells, which may be undergoing neosis, a process of tumor stem cell generation; 2) tracking the effects of cell cycle related toxic agents on cell lines; 3) using MicroRNAs to reverse the polarization state in macrophages to induce tumor cell killing; 4) development of liposomal nanoformulations to overcome Multi-Drug Resistance (MDR) in ovarian cancer cells; and 5) development of dual sensitive micelles to specifically target matrix metalloproteinase 2 (MMP2) over-expressing cell lines. © 2017 International Society for Advancement of Cytometry.
Assuntos
Composição de Medicamentos/métodos , Citometria de Fluxo/tendências , Holografia/tendências , Imagem Molecular/tendências , Resistência a Múltiplos Medicamentos , Humanos , Lipossomos/uso terapêutico , Micelas , Nanotecnologia/tendênciasRESUMO
PURPOSE: The effect of existing anti-cancer therapies is based mainly on the stimulation of apoptosis in cancer cells. Here, we have demonstrated the ability of a catalytically-reactive nanoparticle-based complex of cytochrome c with cardiolipin (Cyt-CL) to induce the apoptosis and killing of cancer cells in a monolayer cell culture. METHODS: Cyt-CL nanoparticles were prepared by complexing CytC with different molar excesses of CL. Following characterization, cytotoxicity and apoptosis inducing effects of nanoparticles were investigated. In an attempt to identify the anticancer activity mechanism of Cyt-CL, pseudo-lipoxygenase and lipoperoxidase reaction kinetics were measured by chemiluminescence. RESULTS: Using chemiluminescence, we have demonstrated that the Cyt-CL complex produces lipoperoxide radicals in two reactions: by decomposition of lipid hydroperoxides, and by lipid peroxidation under the action of H2O2. Antioxidants inhibited the formation of lipid radicals. Cyt-CL nanoparticles, but not the CytC alone, dramatically enhanced the level of apoptosis and cell death in two cell lines: drug-sensitive (A2780) and doxorubicin-resistant (A2780-Adr). The proposed mechanism of the cytotoxic action of Cyt-CL involves either penetration through the cytoplasm and outer mitochondrial membrane and catalysis of lipid peroxidation reactions at the inner mitochondrial membrane, or/and activation of lipid peroxidation within the cytoplasmic membrane. CONCLUSIONS: Here we propose a new type of anticancer nano-formulation, with an action based on the catalytic action of Cyt-CL nanoparticles on the cell membrane and and/or mitochondrial membranes that results in lipid peroxidation reactions, which give rise to activation of apoptosis in cancer cells, including multidrug resistant cells.
Assuntos
Antineoplásicos/farmacologia , Cardiolipinas/farmacologia , Citocromos c/farmacologia , Nanopartículas/química , Antineoplásicos/química , Apoptose , Cardiolipinas/química , Catálise , Técnicas de Cultura de Células , Linhagem Celular Tumoral , Membrana Celular/metabolismo , Sobrevivência Celular , Citocromos c/química , Doxorrubicina/química , Doxorrubicina/farmacologia , Composição de Medicamentos , Resistência a Múltiplos Medicamentos , Resistencia a Medicamentos Antineoplásicos , Humanos , Peróxido de Hidrogênio/química , Peróxido de Hidrogênio/metabolismo , Cinética , Peroxidação de LipídeosRESUMO
Dendrimers are highly branched polymers with easily modifiable surfaces. This makes them promising structures for functionalization and also for conjugation with drugs and DNA/RNA. Their architecture, which can be controlled by different synthesis processes, allows the control of characteristics such as shape, size, charge, and solubility. Dendrimers have the ability to increase the solubility and bioavailability of hydrophobic drugs. The drugs can be entrapped in the intramolecular cavity of the dendrimers or conjugated to their functional groups at their surface. Nucleic acids usually form complexes with the positively charged surface of most cationic dendrimers and this approach has been extensively employed. The presence of functional groups in the dendrimer's exterior also permits the addition of other moieties that can actively target certain diseases and improve delivery, for instance, with folate and antibodies, now widely used as tumor targeting strategies. Dendrimers have been investigated extensively in the medical field, and cancer treatment is one of the greatest areas where they have been most used. This review will consider the main types of dendrimer currently being explored and how they can be utilized as drug and gene carriers and functionalized to improve the delivery of cancer therapy.
Assuntos
Dendrímeros/química , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos , Técnicas de Transferência de Genes , Nanopartículas , Ácidos Nucleicos , Aminoácidos/química , Animais , Antineoplásicos/administração & dosagem , Dendrímeros/síntese química , Portadores de Fármacos/síntese química , Humanos , Peso Molecular , Nanomedicina , Neoplasias/tratamento farmacológico , Ácidos Nucleicos/genética , Tamanho da PartículaRESUMO
Dual stimuli-sensitive mixed polymeric micelles (MM) are developed for co-delivery of the endogenous tumor suppressor miRNA-34a and the chemotherapeutic agent doxorubicin (Dox) into cancer cells. The novelty of the system resides in two stimuli-sensitive prodrugs, a matrix metalloproteinase 2 (MMP2)-sensitive Dox conjugate and a reducing agent (glutathione, GSH)-sensitive miRNA-34a conjugate, self-assembled in a single particle decorated with a polyethylene glycol corona for longevity, and a cell-penetrating peptide (TATp) for enhanced intracellular delivery. The MMP2-sensitivity of the system results in threefold higher cytotoxicity in MMP2-overexpressing HT1080 cells compared to low MMP2-expressing MCF7 cells. Cellular internalization of Dox increases by more than 70% after inclusion of TATp to the formulation. MMP2-sensitive MM also inhibits proliferation and migration of HT1080 cells. Moreover, GSH-sensitive MM allows for an efficient downregulation of Bcl2, survivin, and notch1 (65%, 55%, and 46%, respectively) in HT1080 cells. Combination of both conjugates in dual sensitive MM reduces HT1080 cell viability to 40% and expression of Bcl2 and survivin. Finally, 50% cell death is observed in 3D models of tumor mass. The results confirm the potential of the MM to codeliver miRNA-34a and doxorubicin triggered by dual stimuli inherent of tumor tissues.
Assuntos
Doxorrubicina/uso terapêutico , Sistemas de Liberação de Medicamentos , Técnicas de Transferência de Genes , Micelas , MicroRNAs/administração & dosagem , Neoplasias/tratamento farmacológico , Tamanho da Partícula , Polímeros/química , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Doxorrubicina/farmacologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Glutationa/metabolismo , Humanos , Metaloproteinase 2 da Matriz/metabolismo , MicroRNAs/metabolismo , Neoplasias/genética , Neoplasias/patologia , Esferoides Celulares/efeitos dos fármacos , Esferoides Celulares/metabolismoRESUMO
PURPOSE: To develop transferrin (Tf)-targeted delivery systems for the pro-apoptotic drug, NCL-240, and to evaluate the efficacy of this delivery system in ovarian cancer NCI/ADR-RES cells, grown in vitro in a 3D spheroid model. METHODS: Tf-targeted PEG-PE-based micellar and ePC/CHOL-based liposomal delivery systems for NCL-240 were prepared. NCI/ADR-RES cells were used to generate spheroids by a non-adhesive liquid overlay technique. Spheroid growth and development were monitored by size (diameter) analysis and H&E staining. The targeted formulations were compared to untargeted ones in terms of their degree of spheroid association and penetration. A cell viability analysis with NCL-240-loaded micelles and liposomes was performed to assess the effectiveness of Tf-targeting. RESULTS: Tf-targeted polymeric micelles and Tf-targeted liposomes loaded with NCL-240 were prepared. NCI/ADR-RES cells generated spheroids that demonstrated the presence of a distinct necrotic core along with proliferating cells in the spheroid periphery, partly mimicking in vivo tumors. The Tf-targeted micelles and liposomes had a deeper spheroid penetration as compared to the untargeted delivery systems. Cell viability studies using the spheroid model demonstrated that Tf-mediated targeting markedly improved the cytotoxicity profile of NCL-240. CONCLUSION: Transferrin targeting enhanced delivery and effectiveness of micelles and liposomes loaded with NCL-240 against NCI/ADR-RES cancer cells in a 3D spheroid model.
Assuntos
Apoptose/efeitos dos fármacos , Clorofenóis/administração & dosagem , Sistemas de Liberação de Medicamentos/métodos , Micelas , Neoplasias Ovarianas , Pró-Fármacos/administração & dosagem , Triazóis/administração & dosagem , Apoptose/fisiologia , Linhagem Celular Tumoral , Clorofenóis/metabolismo , Feminino , Humanos , Lipossomos , Neoplasias Ovarianas/tratamento farmacológico , Neoplasias Ovarianas/metabolismo , Pró-Fármacos/metabolismo , Transferrina/administração & dosagem , Transferrina/metabolismo , Triazóis/metabolismo , Células Tumorais CultivadasRESUMO
In response to the challenges of cancer chemotherapeutics, including poor physicochemical properties, low tumor targeting, insufficient tumor cell internalization/bioavailability, and side effects, we developed a unique tumor-targeted micellar drug-delivery platform. Using paclitaxel as a model therapeutic, a nanopreparation composed of a matrix metalloproteinase 2 (MMP2)-sensitive self-assembly PEG 2000-paclitaxel conjugate (as a prodrug and MMP 2-sensitive moiety), transactivating transcriptional activator peptide-PEG1000-phosphoethanolamine (PE) (a cell-penetrating enhancer), and PEG1000-PE (a nanocarrier building block) was prepared. Several major drug delivery strategies, including self-assembly, PEGylation, the enhanced permeability and retention effect, stimulus sensitivity, a cell-penetrating moiety, and the concept of prodrug, were used in design of this nanoparticle in a collaborative manner. The nanopreparation allowed superior cell internalization, cytotoxicity, tumor targeting, and antitumor efficacy in vitro and in vivo over its nonsensitive counterpart, free paclitaxel and conventional micelles. This uniquely engineered nanoparticle has potential for effective intracellular delivery of drug into cancer cells.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Peptídeos Penetradores de Células/farmacologia , Sistemas de Liberação de Medicamentos/métodos , Metaloproteinase 2 da Matriz/metabolismo , Nanopartículas , Neoplasias , Paclitaxel/farmacologia , Polietilenoglicóis/farmacologia , Pró-Fármacos/farmacologia , Animais , Antineoplásicos Fitogênicos/química , Linhagem Celular Tumoral , Peptídeos Penetradores de Células/química , Humanos , Camundongos , Neoplasias/tratamento farmacológico , Neoplasias/enzimologia , Paclitaxel/química , Polietilenoglicóis/química , Pró-Fármacos/química , RatosRESUMO
Multidrug resistance (MDR) mediated by P-glycoprotein overexpression in solid tumors is a major factor in the failure of many forms of chemotherapy. Here we evaluated phospholipid-modified, low-molecular-weight polyethylenimine (DOPE-PEI) nanocarriers for intravenous delivery of anti-P-pg siRNA to tumors with the final goal of modulating MDR in breast cancer. First, we studied the biodistribution of DOPE-PEI nanocarriers and the effect of PEG coating in a subcutaneous breast tumor model. Four hours postinjection, PEGylated carriers showed an 8% injected dose (ID) accumulation in solid tumor via the enhanced permeability and retention effect and 22% ID in serum due to a prolonged, PEG-mediated circulation. Second, we established the therapeutic efficacy and safety of DOPE-PEI/siRNA-mediated P-gp downregulation in combination with doxorubicin (Dox) chemotherapy in MCF-7/MDR xenografts. Weekly injection of siRNA nanopreparations and Dox for up to 5 weeks sensitized the tumors to otherwise non-effective doses of Dox and decreased the tumor volume by threefold vs controls. This therapeutic improvement in response to Dox was attributed to the significant, sequence-specific P-gp downregulation in excised tumors mediated by the DOPE-PEI formulations.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Resistência a Múltiplos Medicamentos , Resistencia a Medicamentos Antineoplásicos , Neoplasias/tratamento farmacológico , Fosfolipídeos/química , Polietilenoimina/química , RNA Interferente Pequeno/administração & dosagem , Subfamília B de Transportador de Cassetes de Ligação de ATP/metabolismo , Animais , Linhagem Celular Tumoral , Doxorrubicina/administração & dosagem , Sistemas de Liberação de Medicamentos , Feminino , Xenoenxertos , Camundongos Endogâmicos BALB C , Nanopartículas/administração & dosagem , Fosfolipídeos/administração & dosagem , Polietilenoimina/administração & dosagemRESUMO
Gene therapy represents a potential efficient approach of disease prevention and therapy. However, due to their poor in vivo stability, gene molecules need to be associated with delivery systems to overcome extracellular and intracellular barriers and allow access to the site of action. Cationic polymeric nanoparticles are popular carriers for small interfering RNA (siRNA) and DNA-based therapeutics for which efficient and safe delivery are important factors that need to be optimized. Micelle-like nanoparticles (MNP) (half micelles, half polymeric nanoparticles) can overcome some of the disadvantages of such cationic carriers by unifying in one single carrier the best of both delivery systems. In this review, we will discuss how the unique properties of MNP including self-assembly, condensation and protection of nucleic acids, improved cell association and gene transfection, and low toxicity may contribute to the successful application of siRNA- and DNA-based therapeutics into the clinic. Recent developments of MNP involving the addition of stimulus-sensitive functions to respond specifically to pathological or externally applied "triggers" (e.g., temperature, pH or enzymatic catalysis, light, or magnetic fields) will be discussed. Finally, we will overview the use of MNP as two-in-one carriers for the simultaneous delivery of different agents (small molecules, imaging agents) and nucleic acid combinations.
Assuntos
DNA/química , Portadores de Fármacos/química , Micelas , Nanopartículas/química , RNA Interferente Pequeno/química , Animais , DNA/administração & dosagem , Portadores de Fármacos/administração & dosagem , Humanos , Nanopartículas/administração & dosagem , RNA Interferente Pequeno/administração & dosagemRESUMO
In this study, transferrin (Tf)-modified poly(ethylene glycol)-phosphatidylethanolamine (mPEG-PE) micelles loaded with the poorly water-soluble drug, R547 (a potent and selective ATP-competitive cyclin-dependent kinase (CDK) inhibitor), were prepared and evaluated for their targeting efficiency and cytotoxicity in vitro and in vivo to A2780 ovarian carcinoma cells, which overexpress transferrin receptors (TfR). At 10 mM lipid concentration, both Tf-modified and plain micelles solubilized 800 µg of R547. Tf-modified micelles showed enhanced interaction with A2780 ovarian carcinoma cells in vitro. The involvement of TfR in endocytosis of Tf-modified micelles was confirmed by colocalization studies of micelle-treated cells with the endosomal marker Tf-Alexa488. We confirmed endocytosis of micelles in an intact form with micelles loaded with a fluorescent dye and additionally labeled with fluorescent lipid. The in vitro cytotoxicity and in vivo tumor growth inhibition studies in A2780-tumor bearing mice confirmed the enhanced efficacy of Tf-modified R547-loaded micelles compared to free drug solution and to nonmodified micelles. The results of this study demonstrate the potential application of Tf-conjugated polymeric micelles in the treatment of tumors overexpressing TfR.