RESUMO
We identified that the chemical linkage of the anticancer drug doxorubicin onto squalene, a natural lipid precursor of the cholesterol's biosynthesis, led to the formation of squalenoyl doxorubicin (SQ-Dox) nanoassemblies of 130-nm mean diameter, with an original "loop-train" structure. This unique nanomedicine demonstrates: (i) high drug payload, (ii) decreased toxicity of the coupled anticancer compound, (iii) improved therapeutic response, (iv) use of biocompatible transporter material, and (v) ease of preparation, all criteria that are not combined in the currently available nanodrugs. Cell culture viability tests and apoptosis assays showed that SQ-Dox nanoassemblies displayed comparable antiproliferative and cytotoxic effects than the native doxorubicin because of the high activity of apoptotic mediators, such as caspase-3 and poly(ADP-ribose) polymerase. In vivo experiments have shown that the SQ-Dox nanomedicine dramatically improved the anticancer efficacy, compared with free doxorubicin. Particularly, the M109 lung tumors that did not respond to doxorubicin treatment were found inhibited by 90% when treated with SQ-Dox nanoassemblies. SQ-Dox nanoassembly-treated MiaPaCa-2 pancreatic tumor xenografts in mice decreased by 95% compared with the tumors in the saline-treated mice, which was significantly higher than the 29% reduction achieved by native doxorubicin. Concerning toxicity, SQ-Dox nanoassemblies showed a fivefold higher maximum-tolerated dose than the free drug, and moreover, the cardiotoxicity study has evidenced that SQ-Dox nanoassemblies did not cause any myocardial lesions, such as those induced by the free doxorubicin treatment. Taken together, these findings demonstrate that SQ-Dox nanoassemblies make tumor cells more sensitive to doxorubicin and reduce the cardiac toxicity, thus providing a remarkable improvement in the drug's therapeutic index.
Assuntos
Doxorrubicina/química , Doxorrubicina/farmacologia , Conformação Molecular , Nanomedicina/métodos , Esqualeno/química , Análise de Variância , Animais , Apoptose/efeitos dos fármacos , Apoptose/fisiologia , Western Blotting , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Microscopia Crioeletrônica , Doxorrubicina/metabolismo , Doxorrubicina/farmacocinética , Feminino , Fluorescência , Coração/anatomia & histologia , Coração/efeitos dos fármacos , Humanos , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Nus , Microscopia Eletrônica de Transmissão , Estrutura Molecular , Ratos , Esqualeno/metabolismo , Troponina T/sangueRESUMO
This review is aimed at combining the published data on taxane formulations into a generalized Drug Delivery approach, starting from the physicochemistry and assessing its relationships with the pharmacokinetics, the biodistribution and the pharmacodynamics. Owing to the number and variety of taxane formulation designs, we considered this class of cytotoxic anticancer agents of particular interest to illustrate the concepts attached to this approach. According to the history of taxane development, we propose a classification as (i) "surfactant-based formulations" first generation, (ii) "surfactant-free formulations" second generation and (iii) "modulated pharmacokinetics drug delivery systems" third generation. Since our objective was to make the link between (i) the physicochemistry of the drug and carrier and (ii) the efficacy and safety of the drug in preclinical animal models and (iii) in human, we focused on the drug delivery technologies that were tested in clinic.
Assuntos
Antineoplásicos/administração & dosagem , Sistemas de Liberação de Medicamentos , Taxoides/administração & dosagem , Animais , Antineoplásicos/farmacocinética , Antineoplásicos/farmacologia , Química Farmacêutica , Portadores de Fármacos/química , Desenho de Fármacos , Humanos , Neoplasias/tratamento farmacológico , Neoplasias/patologia , Tensoativos/química , Taxoides/farmacocinética , Taxoides/farmacologia , Distribuição TecidualRESUMO
Nanotechnology has been considered for the improved delivery of various therapeutic agents, including drugs and genes. Indeed, liposomes and nanoparticles equipped with homing devices for the targeting of receptors over-expressed on the hepatic tissue have improved the treatment of various liver diseases. In this review, various nanotechnology approaches employed for the treatment/imaging of liver disease, either in preclinical or in clinic are discussed.
Assuntos
Hepatopatias/diagnóstico , Hepatopatias/terapia , Nanotecnologia , Animais , Carcinoma Hepatocelular/diagnóstico , Carcinoma Hepatocelular/terapia , Diagnóstico por Imagem , Sistemas de Liberação de Medicamentos , Hepatite/terapia , Humanos , Neoplasias Hepáticas/diagnóstico , Neoplasias Hepáticas/terapia , Nanopartículas de MagnetitaRESUMO
Development of chemoresistance and rapid inactivation of gemcitabine (Gem), the standard therapy for advanced pancreatic cancer, are responsible of the major therapeutic failures. To overcome the above drawbacks we designed a novel nanomedicine strategy for Gem nanoparticle (NP) formulation based on squalene conjugation. The purpose was to investigate the antitumor efficacy of gemcitabine-squalene (SQ-Gem) NPs on chemoresistant and chemosensitive pancreatic adenocarcinoma models. Cell viability and apoptosis assays showed that SQ-Gem NPs displayed higher antiproliferative and cytotoxic effects, particularly in chemoresistant Panc1 tumor cells. In in vivo studies, compared to native Gem, SQ-Gem NPs decreased significantly the tumor growth, prevented tumor cell invasion, and prolonged the survival time of mice bearing orthotopic pancreatic tumors. These results correlate with a greater reduction of Ki-67 and induction of apoptosis. These findings demonstrate the feasibility of utilizing SQ-Gem NPs to make tumor cells more sensitive to Gem and thus provide an efficient new therapeutic alternative for pancreatic adenocarcinoma. FROM THE CLINICAL EDITOR: Pancreatic malignancies represent some of the most notoriously treatment resistant cancer varieties. This paper discusses a novel and promising nanotechnology-based treatment approach, currently at the basic science stage.
Assuntos
Adenocarcinoma/tratamento farmacológico , Antimetabólitos Antineoplásicos/uso terapêutico , Desoxicitidina/análogos & derivados , Nanopartículas/química , Neoplasias Pancreáticas/tratamento farmacológico , Esqualeno/uso terapêutico , Adenocarcinoma/patologia , Animais , Antimetabólitos Antineoplásicos/administração & dosagem , Antimetabólitos Antineoplásicos/química , Apoptose/efeitos dos fármacos , Linhagem Celular , Linhagem Celular Tumoral , Desoxicitidina/administração & dosagem , Desoxicitidina/química , Desoxicitidina/uso terapêutico , Feminino , Humanos , Camundongos , Camundongos Nus , Nanomedicina , Pâncreas/efeitos dos fármacos , Pâncreas/patologia , Neoplasias Pancreáticas/patologia , Esqualeno/administração & dosagem , Esqualeno/química , GencitabinaRESUMO
This study reports the design of a novel theragnostic nanomedicine which combines (i) the ability to target a prodrug of gemcitabine to an experimental solid tumor under the influence of a magnetic field with (ii) the imaging of the targeted tumoral nodule. This concept is based on the inclusion of magnetite nanocrystals into nanoparticles (NPs) constructed by self-assembling molecules of the squalenoyl gemcitabine (SQgem) bioconjugate. The nanocomposites are characterized by an unusually high drug loading, a significant magnetic susceptibility, and a low burst release. When injected to the L1210 subcutaneous mice tumor model, these magnetite/SQgem NPs were magnetically guided, and they displayed considerably greater anticancer activity than the other anticancer treatments (magnetite/SQgem NPs nonmagnetically guided, SQgem NPs, or gemcitabine free in solution). The histology and immunohistochemistry investigation of the tumor biopsies clearly evidenced the therapeutic superiority of the magnetically guided nanocomposites, while Prussian blue staining confirmed their accumulation at the tumor periphery. The superior therapeutic activity and enhanced tumor accumulation has been successfully visualized using T(2)-weighted imaging in magnetic resonance imaging (MRI). This concept was further enlarged by (i) the design of squalene-based NPs containing the T(1) Gd(3+) contrast agent instead of magnetite and (ii) the application to other anticancer squalenoyls, such as, cisplatin, doxorubicin, and paclitaxel. Thus, by combining different anticancer medicines as well as contrast imaging agents in NPs, we open the door toward generic conceptual framework for cancer treatment and diagnosis. This new theragnostic nanotechnology platform is expected to have important applications in cancer therapy.
Assuntos
Nanocompostos/química , Nanomedicina/métodos , Neoplasias/diagnóstico , Neoplasias/tratamento farmacológico , Esqualeno/química , Animais , Antineoplásicos/química , Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Linhagem Celular Tumoral , Preparações de Ação Retardada , Desoxicitidina/análogos & derivados , Desoxicitidina/química , Desoxicitidina/metabolismo , Desoxicitidina/farmacologia , Desoxicitidina/uso terapêutico , Sistemas de Liberação de Medicamentos , Interações Hidrofóbicas e Hidrofílicas , Imageamento por Ressonância Magnética , Magnetismo , Nanopartículas de Magnetita/química , Camundongos , Neoplasias/metabolismo , Pró-Fármacos/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto , GencitabinaRESUMO
Gemcitabine, an anticancer nucleoside analogue, undergoes rapid enzymatic degradation following intravenous injection. This necessitates the administration of a high order of doses to observe a required therapeutic response, while such high doses result in significant side effects. To improve the intravenous delivery of gemcitabine and simultaneously enhance its antitumor activity, we have investigated its incorporation into a drug nanoplatform based on the biodegradable polymer chitosan. Two gemcitabine loading methods have been investigated: (i) entrapment into the polymeric network (entrapment procedure): drug incorporation prior to the coacervation process that leads to the formation of gemcitabine-loaded chitosan (GemChit) nanoparticles; and (ii) surface deposition onto already formed chitosan nanoparticles after incubation in gemcitabine solution (adsorption procedure). The former method produced much higher gemcitabine loading values and a sustained release profile. The main factors determining the gemcitabine loading and release kinetic have also been analyzed. Following intravenous injection, the GemChit formulation displayed a significantly improved antitumor activity comparatively to free gemcitabine, which was further confirmed by histology and immunohistochemistry studies, suggesting the potential of this chitosan-based gemcitabine nanomedicine for the effective treatment of tumors.
Assuntos
Antimetabólitos Antineoplásicos , Quitosana , Desoxicitidina/análogos & derivados , Nanopartículas/química , Neoplasias Experimentais/tratamento farmacológico , Animais , Antimetabólitos Antineoplásicos/química , Antimetabólitos Antineoplásicos/farmacocinética , Antimetabólitos Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Quitosana/química , Quitosana/farmacocinética , Quitosana/farmacologia , Preparações de Ação Retardada , Desoxicitidina/química , Desoxicitidina/farmacocinética , Desoxicitidina/farmacologia , Portadores de Fármacos/química , Portadores de Fármacos/farmacocinética , Portadores de Fármacos/farmacologia , Humanos , Cinética , Camundongos , Camundongos Endogâmicos DBA , GencitabinaRESUMO
A new prodrug of gemcitabine, based on the covalent coupling of squalene to gemcitabine (GemSQ), has been designed to enhance the anticancer activity of gemcitabine, a nucleoside analogue active against a wide variety of tumors. In the present study, the feasibility of encapsulating GemSQ into liposomes either PEGylated or non-PEGylated has been investigated. The in vivo anticancer activity of these formulations has been tested on subcutaneous grafted L1210wt leukemia model and compared to that of free gemcitabine. The liposomal GemSQ appears to be a potential delivery system for the effective treatment of tumors.
Assuntos
Antimetabólitos Antineoplásicos/administração & dosagem , Desoxicitidina/análogos & derivados , Lipossomos/química , Pró-Fármacos/administração & dosagem , Esqualeno/química , Animais , Varredura Diferencial de Calorimetria , Química Farmacêutica , Desoxicitidina/administração & dosagem , Desoxicitidina/química , Leucemia/tratamento farmacológico , Camundongos , Tamanho da Partícula , Polietilenoglicóis/química , Pró-Fármacos/química , Espalhamento a Baixo Ângulo , Difração de Raios X , GencitabinaRESUMO
Using the anticancer compound paclitaxel as a model drug, this study investigates the potential of the squalenoylation technology (i.e., bioconjugation with the natural lipid squalene) in addressing the drug ability and delivery issues of poorly soluble therapeutic agents. In this view, a variety of novel squalene-based prodrugs of the anticancer compound paclitaxel were synthesized, which produced nanoparticles in water. These prodrugs were obtained by covalent coupling of the paclitaxel 2'-hydroxyl group as direct ester, as well as with a succinate or a diglycolate ester as cleavable linker to the 1,1',2-tris-norsqualenoic acid. The hydrophilicity of these paclitaxel bioconjugates was increased by placing poly(ethylene glycol) chains of different lengths between paclitaxel and the squalenoyl moiety. All these prodrugs self-assembled into nanosized aggregates in aqueous solution as characterized by dynamic light scattering, atomic force microscopy, and transmission electron microscopy. The critical aggregation concentration was very low, ranging from 0.09 to 0.4 mg/L. Zeta potential measurements revealed that all squalenoyl-paclitaxel nanoassemblies (NA) held a global negative charge and appeared stable in water for several weeks as determined by particle size measurement. The release of paclitaxel from NA was evaluated in different conditions and in the presence of serum and depended on the nature of the linker used. Preliminary biological assessment showed that these squalenoyl-paclitaxel NA induced the formation of microtubule bundles in HT-29 and KB-31 cells, and additionally displayed notable cytotoxicity on a lung tumor cell line. Furthermore, the cytotoxic activity of these different prodrugs correlated closely with the observed linker stability. Overall, the squalenoylation nanotechnology opens up interesting perspectives for the development of injectable prodrugs of poorly soluble therapeutic compounds by addressing the associated physicochemical and biopharmaceutical challenges.
Assuntos
Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Nanopartículas/química , Paclitaxel/química , Paclitaxel/farmacologia , Pró-Fármacos/farmacologia , Esqualeno/farmacologia , Animais , Antineoplásicos/química , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Desenho de Fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Camundongos , Microtúbulos/química , Conformação Molecular , Tamanho da Partícula , Polietilenoglicóis/química , Pró-Fármacos/síntese química , Pró-Fármacos/química , Esqualeno/química , Estereoisomerismo , Propriedades de Superfície , Fatores de Tempo , Células Tumorais CultivadasRESUMO
4-(N)-1,1',2-trisnor-squalenoyldideoxycytidine monophosphate (SQddC-MP) and 4-(N)-1,1',2-trisnor-squalenoylgemcitabine monophosphate (SQdFdC-MP) were synthesized using phosphoramidite chemistry. These amphiphilic molecules self-assembled to about hundred nanometers size nanoassemblies in aqueous medium. Nanoassemblies of SQddC-MP displayed significant anti-HIV activity whereas SQdFdC-MP nanoassemblies displayed promising anticancer activity on leukemia cells. These results suggested that squalene conjugate of negatively charged nucleotide analogues efficiently penetrated within cells. Thus, we propose a new prodrug strategy for improved delivery of nucleoside analogues to ameliorate their biological efficacy.
Assuntos
Fármacos Anti-HIV/química , Antineoplásicos/química , Nanopartículas/química , Nucleosídeos/química , Pró-Fármacos/química , Esqualeno/química , Animais , Fármacos Anti-HIV/síntese química , Fármacos Anti-HIV/farmacologia , Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Camundongos , Nanopartículas/ultraestrutura , Nucleosídeos/síntese química , Nucleosídeos/farmacologia , Tamanho da Partícula , Pró-Fármacos/síntese química , Pró-Fármacos/farmacologiaRESUMO
Squalene is a natural lipid belonging to the terpenoid family and a precursor of cholesterol biosynthesis. It is synthesized in humans and also in a wide array of organisms and substances, from sharks to olives and even bran, among others. Because of its significant dietary benefits, biocompatibility, inertness, and other advantageous properties, squalene is extensively used as an excipient in pharmaceutical formulations for disease management and therapy. In addition, squalene acts as a protective agent and has been shown to decrease chemotherapy-induced side-effects. Moreover, squalene alone exhibits chemopreventive activity. Although it is a weak inhibitor of tumor cell proliferation, it contributes either directly or indirectly to the treatment of cancer due to its potentiation effect. In addition, squalene enhances the immune response to various associated antigens, and it is therefore being investigated for vaccine delivery applications. Since this triterpene is well absorbed orally, it has been used to improve the oral delivery of therapeutic molecules. All of these qualities have rendered squalene a potentially interesting excipient for pharmaceutical applications, especially for the delivery of vaccines, drugs, genes, and other biological substances. This paper is the first review of its kind and offers greater insight into squalene's direct or indirect contribution to disease management and therapy.
Assuntos
Portadores de Fármacos/administração & dosagem , Neoplasias/prevenção & controle , Esqualeno/administração & dosagem , Administração Oral , Animais , Emulsões/química , Excipientes/administração & dosagem , Técnicas de Transferência de Genes , Humanos , Modelos Moleculares , Estrutura Molecular , Esqualeno/química , Terpenos/química , Vacinas/administração & dosagemRESUMO
Gemcitabine hydrochloride is an anticancer nucleoside analogue indicated in clinic for the treatment of various solid tumors. Although this drug has been demonstrated to display anticancer activity against a wide variety of tumors, it is needed to be administered at high doses to elicit the required therapeutic response, simultaneously leading to severe adverse effects. We hypothesized that the efficient delivery of gemcitabine to tumors using a biodegradable carrier system could reduce the dose required to elicit sufficient therapeutic response. Thus, we have developed a nanoparticle formulation of gemcitabine suitable for parenteral administration based on the biodegradable polymer poly(octylcyanoacrylate) (POCA). The nanoparticles were synthesized by anionic polymerization of the corresponding monomer. Two drug loading methods were analyzed: the first one based on gemcitabine surface adsorption onto the preformed nanoparticles, and the second method being gemcitabine addition before the polymerization process leading to drug entrapment in the polymeric network. A detailed investigation of the capabilities of the polymer particles to load this drug is described. Gemcitabine entrapment into the polymer matrix yielded a higher drug loading and a slower drug release profile as compared with drug adsorption procedure. The main factors determining the gemcitabine incorporation to the polymer network were the nanoparticles preparation procedure, the monomer concentration, the surfactant concentration, the pH, and the drug concentration. The release kinetic of gemcitabine was found to be controlled by the pH and the type of drug incorporation. The cytotoxicity studies performed on L1210 tumor cells revealed a similar anticancer activity of the gemcitabine-loaded POCA (GPOCA) nanoparticle as free gemcitabine. Following intravenous administration into the mice bearing L1210 wt subcutaneous tumor, the GPOCA nanoparticles displayed significantly greater anticancer activity compared to free gemcitabine; this has been additionally confirmed by histology and immunohistochemistry studies, suggesting the potential of GPOCA for the efficient treatment of cancer.
Assuntos
Antimetabólitos Antineoplásicos/administração & dosagem , Desoxicitidina/análogos & derivados , Leucemia L1210/tratamento farmacológico , Nanopartículas , Animais , Antimetabólitos Antineoplásicos/farmacologia , Cianoacrilatos/química , Desoxicitidina/administração & dosagem , Desoxicitidina/farmacologia , Portadores de Fármacos/química , Concentração de Íons de Hidrogênio , Injeções Intravenosas , Camundongos , Camundongos Endogâmicos DBA , Polímeros/química , GencitabinaRESUMO
Gemcitabine (2',2'-difluorodeoxyribofuranosylcytosine) is an anticancer nucleoside analogue active against a wide variety of solid tumors. However, following intravenous administration, this drug is rapidly inactivated by enzymatic deamination and displays a short biological half-life necessitating the administration of high doses leading also to unwanted side effects. To overcome these drawbacks and to improve the therapeutic index of gemcitabine, we have recently developed the concept of squalenoylation which consisted in the bioconjugation of gemcitabine with squalene, a natural lipid. In our preliminary studies, we have shown that this bioconjugate (SQgem) self-organized in water as nanoassemblies with considerable resistance to deamination and significantly higher anticancer activity compared with gemcitabine in an intravenously grafted tumor model in mice. To further establish the candidature of this nanomedicine for clinical trials, in this communication we have tested the preclinical efficacy of squalenoyl gemcitabine nanomedicine on several human tumor cell lines and on the subcutaneously grafted experimental L1210 murine tumor in mice. SQgem nanomedicine displayed an efficient cytotoxicity against a variety of human tumor cell lines in the 60 human tumor cell panel. In vivo, following intravenous administration, SQgem nanomedicine displayed a superior anticancer activity against subcutaneous L1210 tumor, comparatively to gemcitabine. The molecular mechanism behind the anticancer efficacy of SQgem has been investigated by flow cytometry analysis and protein expression profiling of L1210 wt cells treated in vitro with the squalenoyl gemcitabine bioconjugate. It was found that this nanomedicine arrested the cell cycle in G2/M, characterized by an increased cyclin A and cyclin E expression, and activation of caspase-3 and the cleavage of poly(ADP-ribose) polymerase with an increase of cytochrome C level. Taken together, these results suggest that the cell kill by this nanomedicine occurred through mitochondrial apoptotic triggered pathway, similarly to that of gemcitabine free.
Assuntos
Antineoplásicos/química , Antineoplásicos/farmacologia , Desoxicitidina/análogos & derivados , Nanoestruturas/química , Animais , Antineoplásicos/síntese química , Apoptose/efeitos dos fármacos , Caspase 3/metabolismo , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Ciclina A/metabolismo , Ciclina E/metabolismo , Desoxicitidina/síntese química , Desoxicitidina/química , Desoxicitidina/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Leucemia L1210/tratamento farmacológico , Leucemia L1210/metabolismo , Leucemia L1210/patologia , Camundongos , Camundongos Endogâmicos DBA , Nanomedicina , Nanotecnologia , Análise Serial de Proteínas , Esqualeno/análogos & derivados , Esqualeno/química , GencitabinaRESUMO
Gemcitabine is widely used against a variety of solid tumors; however, it possesses some important drawbacks such as rapid deamination leading to short biological half-life and induction of tumor resistance. We have shown previously that the covalent coupling of squalene (a precursor of cholesterol in sterol biosynthesis) to gemcitabine resulted in a potent nanomedicine, squalenoyl gemcitabine (SQdFdC), which displayed appreciable anticancer activity. Now, the present study describes the concept of magnetic responsiveness of SQdFdC nanoparticles obtained by the nanoprecipitation of SQdFdC around magnetite nanoparticles. To investigate these new core/shell nanoparticles, we have compared their structure, chemical composition and surface properties with those of either the magnetic core alone or of the SQdFdC coating material. X-ray diffraction and infrared spectroscopy studies have shown that the composite core/shell particles displayed an intermediate behavior between that of pure magnetite and of pure SQdFdC nanoparticles, whereas dark-field, high-resolution transmission electron microscopy allowed clear demonstration of the core/shell structure. Electrophoresis measurements as a function of both pH and ionic strength, as well as thermodynamic consideration, showed similar behavior of core/shell and pure SQdFdC nanoparticles, suggesting again the coating of the magnetite core by the SQdFdC prodrug. The two important parameters to be controlled in the efficient adsorption of SQdFdC onto magnetite nanocores were the magnetite/SQdFdC weight ratio and the pluronic F-68 concentration. Pluronic F-68 was found to play a key role as a surfactant in the generation of stable composite core/shell nanoparticle suspensions. Finally, the characterization of the magnetic properties of these core/shell nanoparticles revealed that if the squalenoyl shell reduced the magnetic responsiveness of the particles, it kept unchanged their soft ferrimagnetic character. Thus, the heterogeneous structure of these nanoparticles could confer them both magnetic field responsiveness and potential applicability as a drug carrier for active targeting to solid tumors.
Assuntos
Antineoplásicos/química , Desoxicitidina/análogos & derivados , Magnetismo , Nanopartículas/química , Esqualeno/análogos & derivados , Esqualeno/química , Desoxicitidina/química , Elétrons , Cinética , Microscopia Eletrônica de Transmissão , Estrutura Molecular , Nanopartículas/ultraestrutura , Espectrofotometria Infravermelho , Propriedades de Superfície , Termodinâmica , Difração de Raios X , GencitabinaRESUMO
Gemcitabine (2',2'-difluorodeoxyribofuranosylcytosine; dFdC) is an anticancer nucleoside analog active against wide variety of solid tumors. However, this compound is rapidly inactivated by enzymatic deamination and can also induce drug resistance. To overcome the above drawbacks, we recently designed a new squalenoyl nanomedicine of dFdC [4-N-trisnorsqualenoyl-gemcitabine (SQdFdC)] by covalently coupling gemcitabine with the 1,1',2-trisnorsqualenic acid; the resultant nanomedicine displayed impressively greater anticancer activity compared with the parent drug in an experimental murine model. In the present study, we report that SQdFdC nanoassemblies triggered controlled and prolonged release of dFdC and displayed considerably greater t(1/2) (approximately 3.9-fold), mean residence time (approximately 7.5-fold) compared with the dFdC administered as a free drug in mice. It was also observed that the linkage of gemcitabine to the 1,1',2-trisnorsqualenic acid noticeably delayed the metabolism of dFdC into its inactive difluorodeoxyuridine (dFdU) metabolite, compared with dFdC. Additionally, the elimination of SQdFdC nanoassemblies was considerably lower compared with free dFdC, as indicated by lower radioactivity found in urine and kidneys, in accordance with the plasmatic concentrations of dFdU. SQdFdC nanoassemblies also underwent considerably higher distribution to the organs of the reticuloendothelial system, such as spleen and liver (p < 0.05), both after single- or multiple-dose administration schedule. Herein, this paper brings comprehensive pharmacokinetic and biodistribution insights that may explain the previously observed greater efficacy of SQdFdC nanoassemblies against experimental leukemia.
Assuntos
Antineoplásicos/farmacocinética , Desoxicitidina/análogos & derivados , Esqualeno/metabolismo , Animais , Antineoplásicos/sangue , Antineoplásicos/metabolismo , Cromatografia Líquida , Desoxicitidina/sangue , Desoxicitidina/metabolismo , Desoxicitidina/farmacocinética , Feminino , Espectroscopia de Ressonância Magnética , Camundongos , Camundongos Endogâmicos DBA , Espectrometria de Massas em Tandem , Distribuição Tecidual , GencitabinaRESUMO
Univ. Paris-Sud XI, Faculté de Pharmacie, UMR CNRS 8612, 92296 Châtenay-Malabry Cedex, France. patrick.couvreur@u-psud.fr The objective of this review is to discuss the strategies adopted to improve the delivery of gemcitabine to tumors. Concomitant research in this area has implemented a wide variety of approaches such as, aerosolized formulations, prodrug conjugates, liposomes, nanoparticles and beads. Some of these strategies were also aimed at overcoming the rapid metabolization and drug resistance associated with gemcitabine. Aerosolized formulations were employed to treat the local tumors, while other approaches were aimed at the systemic therapy of cancers. The liposomal formulations considerably increased the half-life and the area under the curve (AUC) of gemcitabine, and simultaneously caused a marked improvement in the anticancer activity against experimental solid tumors developed orthotopically or at subcutaneous site. Alternatively, the prodrug conjugates of gemcitabine displayed considerable activity in vivo against various tumors. Especially, in the case of leukemia in which gemcitabine was demonstrated to be inactive, the lipidic conjugates displayed marked efficiency following systemic and oral administration. These conjugates induced greater apoptosis and also caused resistance reversal in the resistant leukemia types. Altogether, the delivery strategies adopted for gemcitabine led to a considerable improvement in the treatment of cancers at the preclinical stage, and some of them are potential candidates for clinical trials.
Assuntos
Antimetabólitos Antineoplásicos/administração & dosagem , Desoxicitidina/análogos & derivados , Neoplasias/tratamento farmacológico , Animais , Antimetabólitos Antineoplásicos/farmacocinética , Ensaios Clínicos como Assunto , Desoxicitidina/administração & dosagem , Desoxicitidina/farmacocinética , Sistemas de Liberação de Medicamentos , Resistencia a Medicamentos Antineoplásicos , Humanos , GencitabinaRESUMO
In this study, the dynamically folded conformation of squalene (SQ) is taken advantage of to link this natural compound to the anticancer nucleoside analogue gemcitabine (gem) in order to achieve the spontaneous formation of nanoassemblies (SQgem) in water. Cryogenic transmission electron microscopy examination reveals particles (104 nm) with a hexagonal or multifaceted shape that display an internal structure made of reticular planes, each particle being surrounded by an external shell. X-ray diffraction evidences the hexagonal molecular packing of SQgem, resulting from the stacking of direct or inverse cylinders. The respective volumes of the gem and SQ molecules as well as molecular modeling of SQgem suggest the stacking of inverse hexagonal phases, in which the central aqueous core, consisting of water and gem molecules, is surrounded by SQ moieties. These SQgem nanoassemblies also exhibit impressively greater anticancer activity than gem against a solid subcutaneously grafted tumor, following intravenous administration. To our knowledge, this is the first demonstration of hexagonal phase organization with a SQ derivative.
Assuntos
Antineoplásicos/química , Nanoestruturas/química , Animais , Antineoplásicos/administração & dosagem , Microscopia Crioeletrônica , Desoxicitidina/administração & dosagem , Desoxicitidina/análogos & derivados , Desoxicitidina/química , Leucemia P388/tratamento farmacológico , Substâncias Macromoleculares/química , Camundongos , Camundongos Endogâmicos DBA , Modelos Moleculares , Nanoestruturas/administração & dosagem , Nanoestruturas/ultraestrutura , Nanotecnologia , Espalhamento a Baixo Ângulo , Esqualeno/análogos & derivados , Esqualeno/química , Difração de Raios X , GencitabinaRESUMO
This study investigates 1) the anticancer efficacy of a new squalenoyl prodrug of gemcitabine (SQgem) in nanoassembly form compared with gemcitabine at equitoxic doses and 2) the subacute and acute preclinical toxicity of these compounds. The toxicity studies revealed that SQgem nanoassemblies, like gemcitabine, were toxic, and they led to dose-dependent mortality after daily i.v. injections for 1 week, irrespective of the route of administration. However, a 4- to 5-day spaced dosing schedule (injections on day 0, 4, 8, and 13) was proved to be safer in terms of weight loss and hematological and other toxicity. Using this spaced dosing schedule, SQgem nanoassemblies exhibited impressive anticancer activity in mice bearing L1210 leukemia because this treatment led to 75% long-term survivors. In contrast, at equitoxic doses, neither free gemcitabine nor cytarabine led to longterm survivors and all the mice of these groups died of the disease. Further toxicity studies performed at lethal doses by blood and serum analysis and organ weight determinations revealed that the hematological toxicity was the dose-limiting toxicity in both SQgem nanoassemblies and gemcitabine, whereas probable gastrointestinal toxicity was also associated with free gemcitabine. The SQgem nanoassemblies did not display hepatotoxicity, which is one of the clinically encountered toxicities of gemcitabine. To summarize, these preclinical studies demonstrated that the toxicological profile of new squalenoyl gemcitabine nanomedicine was not distinct from that of the parent gemcitabine, whereas it was much more potent than gemcitabine at equitoxic doses and cytarabine at clinically relevant doses. These data support the candidature of SQgem for clinical trials.
Assuntos
Antineoplásicos/uso terapêutico , Desoxicitidina/análogos & derivados , Leucemia/tratamento farmacológico , Nanoestruturas/uso terapêutico , Esqualeno/química , Esqualeno/uso terapêutico , Animais , Antineoplásicos/toxicidade , Citarabina/uso terapêutico , Desoxicitidina/química , Desoxicitidina/uso terapêutico , Desoxicitidina/toxicidade , Avaliação Pré-Clínica de Medicamentos , Camundongos , Camundongos Endogâmicos DBA , Nanomedicina , Nanoestruturas/toxicidade , Esqualeno/toxicidade , Resultado do Tratamento , GencitabinaRESUMO
Gemcitabine-squalene is a new prodrug that self-organizes in water forming nanoassemblies. It exhibits better anti-cancer properties in vitro and in vivo than gemcitabine. A liquid chromatography/tandem mass spectrometry assay of gemcitabine-squalene and gemcitabine was developed in human plasma in order to quantitate gemcitabine and its squalene conjugate. After protein precipitation with acetonitrile/methanol (90/10, v/v), the compounds were analyzed by reversed-phase high performance liquid chromatography and detected by tandem mass spectrometry using multiple reaction monitoring. The method was linear over the concentration range of 10-10,000 ng/ml of human plasma for both compounds with an accuracy lower than 10.4% and a precision below 14.8%. The method showed a lower limit of quantitation of 10 ng/ml of human plasma for dFdC and dFdC-SQ. A preliminary in vivo study in mice was shown as application of the method as no significant difference between human and mice plasma for the analysis of dFdC and dFdC-SQ was demonstrated.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Desoxicitidina/análogos & derivados , Esqualeno/sangue , Espectrometria de Massas em Tandem/métodos , Desoxicitidina/sangue , Desoxicitidina/química , Humanos , Estrutura Molecular , Reprodutibilidade dos Testes , Esqualeno/química , GencitabinaRESUMO
Gemcitabine is an anticancer nucleoside analogue active against various solid tumors. However, it possesses important drawbacks like a poor biological half-life and the induction of resistance. With the objective of overcoming the above drawbacks, we designed a new nanomedicine of gemcitabine and studied its anticancer efficacy against leukemia at preclinic. Gemcitabine has been covalently coupled with 1,1',2-tris-nor-squalenic acid to obtain the new anticancer nanomedicine 4-(N)-Tris-nor-squalenoyl-gemcitabine (SQdFdC NA). The SQdFdC NA exhibited, in comparison to gemcitabine, 3.26- and 3.22-folds higher cytotoxicity respectively, in murine resistant leukemia L1210 10K cells and in human leukemia resistant cell line CEM/ARAC8C. Following intravenous treatment of murine aggressive metastatic leukemia L1210 wt bearing mice, the SQdFdC NA caused significant increase in survival time compared to gemcitabine and also led to long-term survivals, which was not the case after gemcitabine treatment. This was attributed to significantly higher deposition of SQdFdC NA in spleen and liver (P<0.05), the major metastatic organs. In comparison to gemcitabine, SQdFdC NA displayed greater ability to induce S-phase arrest of the cancer cells followed by increased apoptotic induction. Interestingly, like gemcitabine, SQdFdC NA didn't induce appreciable differences in blood parameters even at doses higher than those used for anticancer evaluation. The preclinical data obtained in vitro and in vivo with SQdFdC NA demonstrate that this nanomedicine represents a new therapeutic system for the effective treatment of leukemia.