RESUMEN
With the aid of bioorthogonal chemistry, we demonstrate the fabrication of synthetic dendrimers in situ around living cells. Using tetrazine dienophile and aminooxyl/hydrazide aldehyde chemistries, the density of functional groups on the dendrimers exponentially amplified intensities of fluorescent markers in antibody-targeted live cell imaging. This novel "swarming" approach highlights the power of bioorthogonal chemistry and provides a route to non-natural chemical structures on cells, paving the way for the generation of various artificial cellular nanostructures and scaffolds.
RESUMEN
A polymer scaffold, with multiple reactive centres, was synthesised by RAFT polymerisation and conjugated to the antibody herceptin. A hexahistidine RAFT agent enabled the rapid and simple purification of polymer-protein conjugates, while the tetrazine conjugation strategy allows myriad cargos to be attached and amplified.
Asunto(s)
Resinas Acrílicas/química , Colorantes Fluorescentes/química , Histidina/química , Trastuzumab/química , Resinas Acrílicas/síntesis química , Colorantes Fluorescentes/síntesis química , Estructura Molecular , Norbornanos/químicaRESUMEN
Polymerization reactions conducted inside cells must be compatible with the complex intracellular environment, which contains numerous molecules and functional groups that could potentially prevent or quench polymerization reactions. Here we report a strategy for directly synthesizing unnatural polymers in cells through free radical photopolymerization using a number of biocompatible acrylic and methacrylic monomers. This offers a platform to manipulate, track and control cellular behaviour by the in cellulo generation of macromolecules that have the ability to alter cellular motility, label cells by the generation of fluorescent polymers for long-term tracking studies, as well as generate a variety of nanostructures within cells. It is remarkable that free radical polymerization chemistry can take place within such complex cellular environments. This demonstration opens up a multitude of new possibilities for how chemists can modulate cellular function and behaviour and for understanding cellular behaviour in response to the generation of free radicals.
Asunto(s)
Radicales Libres/química , Polimerizacion/efectos de la radiación , Ácidos Polimetacrílicos/síntesis química , Poliestirenos/síntesis química , Acrilatos/química , Acrilatos/efectos de la radiación , Acrilatos/toxicidad , Citoesqueleto de Actina/efectos de los fármacos , Compuestos de Anilina/química , Compuestos de Anilina/efectos de la radiación , Compuestos de Anilina/toxicidad , Movimiento Celular/efectos de los fármacos , Fluorescencia , Células HeLa , Humanos , Metacrilatos/química , Metacrilatos/efectos de la radiación , Metacrilatos/toxicidad , Propano/análogos & derivados , Propano/química , Propano/efectos de la radiación , Fase S/efectos de los fármacos , Estirenos/química , Estirenos/efectos de la radiación , Estirenos/toxicidad , Rayos Ultravioleta , Compuestos de Vinilo/química , Compuestos de Vinilo/efectos de la radiación , Compuestos de Vinilo/toxicidadRESUMEN
The evaluation of the role of physicochemical properties in the toxicity of nanoparticles is important for the understanding of toxicity mechanisms and for controlling the behavior of nanoparticles. The surface charge of nanoparticles is suggested as one of the key parameters which decide their biological impact. In this study, we synthesized fluorophore-conjugated polystyrene nanoparticles (F-PLNPs), with seven different types of surface functional groups that were all based on an identical core, to evaluate the role of surface charge in the cellular uptake of nanoparticles. Phagocytic differentiated THP-1 cells or non-phagocytic A549 cells were incubated with F-PLNP for 4 h, and their cellular uptake was quantified by fluorescence intensity and confocal microscopy. The amount of internalized F-PLNPs showed a good positive correlation with the zeta potential of F-PLNPs in both cell lines (Pearson's r = 0.7021 and 0.7852 for zeta potential vs. cellular uptake in THP-1 cells and nonphagocytic A549 cells, respectively). This result implies that surface charge is the major parameter determining cellular uptake efficiency, although other factors such as aggregation/agglomeration, protein corona formation, and compositional elements can also influence the cellular uptake partly or indirectly.
RESUMEN
The term electroceutical has been used to describe implanted devices that deliver electrical stimuli to modify biological function. Herein, we describe a new concept in electroceuticals, demonstrating for the first time the electrochemical activation of metal-based prodrugs. This is illustrated by the controlled activation of Pt(iv) prodrugs into their active Pt(ii) forms within a cellular context allowing selectivity and control of where, when and how much active drug is generated.
Asunto(s)
Antineoplásicos/farmacología , Complejos de Coordinación/farmacología , Técnicas Electroquímicas/métodos , Compuestos Organoplatinos/farmacología , Profármacos/farmacología , Antineoplásicos/química , Apoptosis/efectos de los fármacos , Complejos de Coordinación/síntesis química , Complejos de Coordinación/química , Electrodos , Células HCT116 , Humanos , Compuestos Organoplatinos/síntesis química , Compuestos Organoplatinos/química , Oxidación-Reducción , Profármacos/síntesis química , Profármacos/químicaRESUMEN
A homogeneous carbene-based palladium catalyst was conjugated to a cell-penetrating peptide, allowing intracellular delivery of catalytically active Pd complexes that demonstrated bioorthogonal activation of a profluorophore within prostate cancer cells.
RESUMEN
Transition metals have been successfully applied to catalyze non-natural chemical transformations within living cells, with the highly efficient labeling of subcellular components and the activation of prodrugs. In vivo applications, however, have been scarce, with a need for the specific cellular targeting of the active transition metals. Here, we show the design and application of cancer-targeting palladium catalysts, with their specific uptake in brain cancer (glioblastoma) cells, while maintaining their catalytic activity. In these cells, for the first time, two different anticancer agents were synthesized simultaneously intracellularly, by two totally different mechanisms (in situ synthesis and decaging), enhancing the therapeutic effect of the drugs. Tumor specificity of the catalysts together with their ability to perform simultaneous multiple bioorthogonal transformations will empower the application of in vivo transition metals for drug activation strategies.
Asunto(s)
Antineoplásicos/metabolismo , Antineoplásicos/uso terapéutico , Neoplasias Encefálicas/tratamiento farmacológico , Glioblastoma/tratamiento farmacológico , Paladio/química , Profármacos/metabolismo , Profármacos/uso terapéutico , Neoplasias Encefálicas/metabolismo , Catálisis , Línea Celular Tumoral , Sistemas de Liberación de Medicamentos , Colorantes Fluorescentes/química , Glioblastoma/metabolismo , Humanos , Nanopartículas del Metal/química , Microscopía Fluorescente , Oligopéptidos/metabolismoRESUMEN
The copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction has proven to be a pivotal advance in chemical ligation strategies with applications ranging from polymer fabrication to bioconjugation. However, application inâ vivo has been limited by the inherent toxicity of the copper catalyst. Herein, we report the application of heterogeneous copper catalysts in azide-alkyne cycloaddition processes in biological systems ranging from cells to zebrafish, with reactions spanning from fluorophore activation to the first reported inâ situ generation of a triazole-containing anticancer agent from two benign components, opening up many new avenues of exploration for CuAAC chemistry.
Asunto(s)
Alquinos/química , Antineoplásicos/química , Azidas/química , Química Clic/métodos , Reacción de Cicloadición/métodos , Colorantes Fluorescentes/química , Triazoles/química , Alquinos/síntesis química , Animales , Antineoplásicos/síntesis química , Azidas/síntesis química , Catálisis , Línea Celular , Cobre , Colorantes Fluorescentes/síntesis química , Células HeLa , Humanos , Nanopartículas del Metal/química , Triazoles/síntesis química , Pez CebraRESUMEN
As a novel prodrug activation strategy Pd(0) nanoparticles, entrapped within a modular polymeric support, were used in cell culture, to synthesise the anticancer agent PP-121 from two non-toxic precursors, thereby inducing cell death in the first example of in situ mediated drug synthesis.
Asunto(s)
Antineoplásicos/síntesis química , Nanopartículas del Metal/química , Paladio/química , Pirazoles/síntesis química , Pirimidinas/síntesis química , Antineoplásicos/química , Antineoplásicos/farmacología , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Estructura Molecular , Tamaño de la Partícula , Pirazoles/química , Pirazoles/farmacología , Pirimidinas/química , Pirimidinas/farmacología , Relación Estructura-Actividad , Propiedades de Superficie , Células Tumorales CultivadasRESUMEN
This Article reports the synthesis, structural characterization, electrochemistry, and photophysical investigations of five groups of luminescent Pt(II) alkynyl complexes bearing N-heterocyclic carbene (NHC) ligands with varying electronic properties. Complexes of the type [Pt(pmdb)(C≡CR)2] 1a-c, [Pt(pm2tz)(C≡CR)2] 2a-d, [Pt(pm3tz)(C≡CR)2] 3a-c, [Pt(ppim)(C≡CR)2] 4(a, b, e), and [Pt(ppbim)(C≡CR)2] 5(a, b, e), where pmdb =1,1'-dipentyl-3,3'-methylene-dibenzimidazoline-2,2'-diylidene, pm2tz = 1,1'-dipentyl-3,3'-methylene-di-1,2,4-triazoline-5,5'-diylidene, pm3tz = 1,1'-dipentyl-3,3'-methylene-di-1,3,4-triazoline-5,5'-diylidene, ppim = 3-pentyl-1-picolylimidazoline-2-ylidene, and ppbim = 3-pentyl-1-picolylbenzimidazoline-2-ylidene, and R = 4-C6H4F, C6H5, 4-C6H4OMe, SiMe3, and 4-C6H4N(C6H5)2, were prepared, and the consequences of the electronic properties of the NHC ligands on the phosphorescent emission efficiencies were studied. Moreover, the emission quantum efficiencies of the previously reported complexes [Pt(pmim)(C≡CR)2] where pmim = 1,1'-dipentyl-3,3'-methylene-diimidazoline-2,2'-diylidene and R = 4-C6H4F 6a, C6H5 6b, and 4-C6H4OMe 6c were also recorded in neat solid and in 10 wt % PMMA film. The square planar coordination geometry with the alkynyl ligands in cis configuration was corroborated for selected complexes by single crystal X-ray diffraction studies. The observed moderate difference in emission efficiencies of the bis-carbene complexes 6a-c, 1a-c, 2a-c, and 3a-c in conjunction with the decreasing electron-donating nature of the NHC ligands, pmim > pmdb > pm2tz ≈ pm3tz, can be attributed to the slight modification of the triplet emission parentage among the different complexes. The quantum efficiencies of complexes 4(a, b) and 5(a, b) bearing pyridyl-NHC ligand were significantly low in comparison to the bis-carbene complexes owing to the significant change in the charge transfer character of the triplet manifold. Complexes 4e and 5e bearing diarylamine phenylacetylenes display high Ïem of 27% and 33% in 10 wt % PMMA film, respectively.