RESUMO
Rotaxane dendrimers with hyperbranched macromolecular interlocked structures and size modulation capacity demonstrate drug binding and release ability upon external stimuli. Mass spectrometry imaging (MSI) can offer the high-throughput screening of endogenous/exogenous compounds. Herein, we reported a novel method to display the in situ spatial distribution of label-free monodispersed type III rotaxane dendrimers (RDs) G1 (first generation, size â¼1.5 nm) and G2 (second generation, size â¼5 nm) that were explored as potential drug vehicles in spleen tissue by using matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-MSI). Experimental results indicated that the trans-2-[3-(4-tert-butylphenyl)-2-methyl-2-propenylidene]malononitrile (DCTB) matrix exhibited the best performance for monodispersed type III RDs G1 and G2. The optimized method was successfully applied to map the in vivo spatial distribution of type III RDs G1 and G2 in the spleen from intraperitoneally injected mice. The MALDI-MSI images revealed that RDs G1 and G2 were relatively stable in the spleen within 24 h after administration. It was found that the identified type III RDs G1 and G2 penetrated through the tunica serosa and were predominantly localized in red pulp regions of spleens. They were also mapped in a marginal zone of spleens simultaneously. There was almost no toxicity of type III RDs G1 and G2 to mice spleens from the H&E results. Furthermore, the type III RDs did not induce the expression of inflammatory cytokines from peripheral blood mononuclear cells (PBMCs) or THP-1 monocytes. The MSI analysis not only demonstrated its ability to image select rotaxane dendrimers in a rapid and efficient manner but also provided tremendous assistance on the applications of the further treatment of cancerous tissue as safe drug carriers. Furthermore, the new strategy demonstrated in this study could be applied on other label-free mechanically interlocked molecules, molecular machines, and macromolecules, which opened a new path to evaluate the toxicological and pharmacokinetic characteristics of these novel materials at the suborgan level.
Assuntos
Dendrímeros/análise , Portadores de Fármacos/análise , Rotaxanos/análise , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Animais , Dendrímeros/farmacocinética , Portadores de Fármacos/farmacocinética , Camundongos , Rotaxanos/farmacocinética , Baço/metabolismo , Distribuição TecidualRESUMO
Major objectives in nanomedicine and nanotherapy include the ability to trap therapeutic molecules inside of nano-carriers, carry therapeutics to the site of the disease with no leakage, release high local concentrations of drug, release only on demand - either autonomous or external, and kill the cancer cells or an infectious organism. This review will focus on mesoporous silica nanoparticle carriers (MSN) with a large internal pore volume suitable for carrying anticancer and antibiotic drugs, and supramolecular components that function as caps that can both trap and release the drugs on-command. Caps that are especially relevant to this review are rotaxanes and pseudorotaxanes that consist of a long chain-like molecule threaded through a cyclic molecule. Under certain conditions discussed throughout this review, the cyclic molecule can be attracted to one end of the rotaxane and in the presence of a stimulus can slide to the other end. When the thread is attached near the pore opening on MSNs, the sliding cyclic molecule can block the pore when it is near the particle or open it when it slides away. The design, synthesis and operation of supramolecular systems that act as stimuli-responsive pore capping devices that trap and release molecules for therapeutic or imaging applications are discussed. Uncapping can either be irreversible because the cap comes off, or reversible when the cyclic molecule is prevented from sliding off by a steric barrier. In the latter case the amount of cargo released (the dose) can be controlled. These nanomachines act as valves. Examples of supramolecular systems stimulated by chemical signals (pH, redox, enzymes, antibodies) or by external physical signals (light, heat, magnetism, ultrasound) are presented. Many of the systems have been studied in vitro proving that they are taken up by cancer cells and release drugs and kill the cells when stimulated. Some have been studied in mouse models; after IV injection they shrink tumors or kill intracellular pathogens after stimulation. Supramolecular constructs offer fascinating, highly controllable and biologically compatible platforms for drug delivery.
Assuntos
Antibacterianos/administração & dosagem , Antineoplásicos/administração & dosagem , Compostos de Cálcio , Sistemas de Liberação de Medicamentos , Substâncias Macromoleculares/administração & dosagem , Nanopartículas , Silicatos , Animais , Linhagem Celular , Humanos , Substâncias Macromoleculares/farmacocinética , Substâncias Macromoleculares/farmacologia , Camundongos , Nanomedicina/métodos , Nanomedicina/tendências , Rotaxanos/administração & dosagem , Rotaxanos/farmacocinética , Rotaxanos/farmacologiaRESUMO
The use of natural compounds to construct biomaterials, including delivery system, is an attractive strategy. In the present study, through threading functional α-cyclodextrins onto the conjugated macromolecules of poly(ethylene glycol) (PEG) and natural compound bile acid, glycopolymers of polyrotaxanes with the active targeting ability are obtained. These glycopolymers self-assemble into micelles as evidenced by dynamic light scattering and transmission electron microscopy, in which glucosamine, as an example of targeting groups, is introduced. These micelles after loading doxorubicin (DOX) exhibit the selective recognition with cancer cells 4T1. Meanwhile, the maximal half inhibitory concentration is determined to be ≈2.5 mg L-1 for the DOX-loaded micelles, close to the value of free DOX·HCl (1.9 mg L-1 ). The cumulative release of DOX at pH 5.5 is faster than at pH 7.4, which may be used as the controlled release system. This drug delivery system assembled by glycopolymers features high drug loading of DOX, superior biocompatibility. The strategy not only utilizes the micellization induced by bile acids, but also overcomes the major limitation of PEG such as the lack of targeting groups. In particular, this drug delivery platform can extend to grafting the other targeting groups, rendering this system more versatile.
Assuntos
Ácidos e Sais Biliares , Doxorrubicina , Portadores de Fármacos , Micelas , Neoplasias Experimentais/tratamento farmacológico , Rotaxanos , Animais , Ácidos e Sais Biliares/química , Ácidos e Sais Biliares/farmacocinética , Ácidos e Sais Biliares/farmacologia , Linhagem Celular Tumoral , Doxorrubicina/química , Doxorrubicina/farmacocinética , Doxorrubicina/farmacologia , Portadores de Fármacos/química , Portadores de Fármacos/farmacocinética , Portadores de Fármacos/farmacologia , Camundongos , Células NIH 3T3 , Neoplasias Experimentais/metabolismo , Neoplasias Experimentais/patologia , Coelhos , Rotaxanos/química , Rotaxanos/farmacocinética , Rotaxanos/farmacologiaRESUMO
Polyrotaxane (PRX) is a promising supramolecular carrier for gene delivery. Classic PRX exhibits a linear structure in which the amine-functionalized α-cyclodextrin (CD) is threaded along the entire polyethylene glycol (PEG) backbone. While promising in vitro, the absence of free PEG moieties after CD threading compromised the in vivo implementation, due to the unfavorable pharmacokinetics (PK) and biodistribution profile. Herein, we developed a multi-arm PRX nanocarrier platform, which has been designed for protective nucleic acid encapsulation, augmented biodistribution and PK, and suitable for intravenous (IV) administration. A key design was to introduce cationic CD rings onto a multi-arm PEG backbone in a spatially selective fashion. The optimal structural design was obtained through iterative rounds of experimentation to determine the appropriate type and density of cationic charge on CD ring, the degree of PEGylation, the size and structure of polymer backbone, etc. This allowed us to effectively deliver large size reporter and therapeutic plasmids in cancer mouse models. Post IV injection, we demonstrated that our multi-arm polymer design significantly enhanced circulatory half-life and PK profile compared to the linear PRX. We continued to use the multi-arm PRX to formulate a therapeutic plasmid encoding an immunomodulatory cytokine, IL-12. When tested in a colon cancer syngeneic mouse model with same background, the IL-12 plasmid was protected by the multi-arm PRX and delivered through the tail vein to the tumor site, leading to a significant tumor inhibition effect. Moreover, our delivery system was devoid of major systemic toxicity.
Assuntos
Ciclodextrinas/química , Portadores de Fármacos/química , Nanopartículas/química , Plasmídeos/administração & dosagem , Poloxâmero/química , Rotaxanos/química , Imunidade Adaptativa/efeitos dos fármacos , Animais , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Ciclodextrinas/farmacocinética , Feminino , Técnicas de Transferência de Genes , Interleucina-12/metabolismo , Camundongos Endogâmicos C57BL , Poloxâmero/farmacocinética , Rotaxanos/farmacocinética , Distribuição Tecidual/efeitos dos fármacos , alfa-Ciclodextrinas/químicaRESUMO
Due to the potential use as transfecting agents of nucleic acids (DNA or RNA), multivalent cationic non-viral vectors have received special attention in the last decade. Much effort has been addressed to synthesize more efficient and biocompatible gene vectors able to transport nucleic acids into the cells without provoking an immune response. Among them, the mostly explored to compact and transfect nucleic acids are: (a) gemini and multivalent cationic lipids, mixed with a helper lipid, by forming lipoplexes; and (b) cationic polymers, polycations, and polyrotaxanes, by forming polyplexes. This review is focused on the progress and recent advances experimented in this area, mainly during the present decade, devoting special attention to the lipoplexes and polyplexes, as follows: (a) to its biophysical characterization (mainly electrostatics, structure, size and morphology) using a wide variety of experimental methods; and (b) to its biological activity (transfection efficacy and cytotoxicity) addressed to confirm the optimum formulations and viability of these complexes as very promising gene vectors of nucleic acids in nanomedicine.
Assuntos
Técnicas de Transferência de Genes , Terapia Genética/métodos , Vetores Genéticos/química , Ácidos Nucleicos/uso terapêutico , Animais , Doenças Genéticas Inatas/patologia , Doenças Genéticas Inatas/terapia , Vetores Genéticos/farmacocinética , Humanos , Lipídeos/química , Lipídeos/farmacocinética , Neoplasias/genética , Neoplasias/patologia , Neoplasias/terapia , Ácidos Nucleicos/farmacocinética , Poliaminas/química , Poliaminas/farmacocinética , Polieletrólitos , Compostos de Amônio Quaternário/química , Compostos de Amônio Quaternário/farmacocinética , Rotaxanos/química , Rotaxanos/farmacocinéticaRESUMO
In the development of anti-cancer drugs, it is important to yield selective cytotoxicity primarily against tumor tissues. To achieve this goal, the use of a polymer-drug conjugate appears to be appealing, simply because it can take the advantage of the so-called enhanced permeability and retention (EPR) effect due to vascular leak in tumors. Among various types of polymers, polyrotaxane (PR) is an interesting candidate and warrants further consideration. It is a self-assembled polymer made entirely of biocompatible components, by threading alpha-cyclodextrin (alpha-CD) molecules with the poly(ethylene glycol) (PEG) chain. The abundance in functional -OH groups on the CD residues renders PR the capability of carrying a large dose of small anti-tumor agents for delivery. Herein, we presented a novel PR-based delivery system using doxorubicin (DOX) as the model anti-cancer drug. Daunorubicin (DNR) was conjugated to the PR polymer via hydrolysable linkages, and upon hydrolysis, doxorubicin was released as the cytotoxic drug. To facilitate an intracellular uptake by the tumor cells of the PR-DOX conjugates, a cell-penetrating low molecular weight protamine (LMWP) peptide was further attached to the two termini of the PR chain. Using an innovative principle established in our laboratory, such as via the inhibition of the cell-penetrating activity by binding with heparin and reversal of this inhibition by subsequent addition of protamine, cellular uptake of the polymer-drug conjugates could be readily regulated. In this paper, we performed in vitro studies to demonstrate the feasibility of this delivery system. The LMWP-PR-DOX conjugates, which yielded a sustained release of DOX over a period of greater than 4 days, were successfully synthesized. Intracellular uptake of these conjugates by A2780 human ovarian cancer cells and regulation of such uptake by heparin and protamine were confirmed by using the MTT assay and also the confocal microscopy method.
Assuntos
Antineoplásicos/administração & dosagem , Ciclodextrinas/química , Portadores de Fármacos/química , Poloxâmero/química , Protaminas/química , Rotaxanos/química , Antineoplásicos/química , Antineoplásicos/farmacocinética , Linhagem Celular Tumoral , Permeabilidade da Membrana Celular , Sobrevivência Celular/efeitos dos fármacos , Ciclodextrinas/farmacocinética , Daunorrubicina/administração & dosagem , Daunorrubicina/química , Daunorrubicina/farmacocinética , Doxorrubicina/administração & dosagem , Doxorrubicina/química , Doxorrubicina/farmacocinética , Portadores de Fármacos/farmacocinética , Sistemas de Liberação de Medicamentos/métodos , Humanos , Poloxâmero/farmacocinética , Protaminas/farmacocinética , Rotaxanos/farmacocinética , SolubilidadeRESUMO
The development of methods to transport peptides into cells via a passive mechanism would greatly aid in the development of therapeutic agents. We recently demonstrated that an impermeable fluoresceinated pentapeptide enters the cytoplasm and nucleus of COS 7 cells in the presence of a host-[2]rotaxane by a mechanism that does not depend on an active cell-mediated process. In this report, we further investigate the ability of the host-[2]rotaxane to deliver peptides possessing a wide range of polarities (negatively charged, positively charged, polar, and apolar side chains) into live cells. Only in the presence of the host-[2]rotaxane were the Fl-peptides taken up by COS 7 and ES2 cells. Flow cytometry experiments demonstrated that the level of delivery is largely temperature and adenosine 5'-triphosphate (ATP) independent, and the membranes remain intact. Although the level of transport does depend upon the nature of the side chains, it does not correlate with calculated LogD values, indicating that an additional interaction with the host-[2]rotaxane is modifying the permeability properties of the peptide. The amount of Fl-peptides transported from an aqueous phase into a chloroform phase in the presence of the host-[2]rotaxane correlates with the intensity of cellular fluorescence. Extraction and U-tube studies show that the Fl-peptide can be released from its complex with the host-[2]rotaxane into an aqueous phase, and the host-[2]rotaxane can transport a greater than a stoichiometric amount of an Fl-peptide through a CHCl3 layer. These studies demonstrate the utility of the host-[2]rotaxane in delivering peptides of all polarities across a cell membrane.