RESUMEN
In this work we describe not previously explored binding studies on the reversible interaction of benzoxaborole with ligands of medical and pharmaceutical interest such as nucleosidic drugs gemcitabine and capecitabine, as well as the hydrophobic chemotherapeutic doxorubicin. We include functional derivatives of benzoxaborole such as a near infrared fluorescent boronolectine, Cy-Bx, The dynamic covalent interaction in physiological conditions was assessed by spectroscopic techniques yielding moderate to high binding affinities. The cytotoxic activity of the drugs upon conjugation to the boronolectins was evaluated revealing significant influence of the bioconjugation status on the cellular viability. The availability of the conjugate for cellular uptake and localization in the model cancer cell line HeLa was assessed by fluorescence imaging. Benzoxaborole and the fluorescent boronolectin Cy-Bx, proved to be versatile conjugation tools for 1,2 and 1,3-diol containing pharmacophores as well as bioisosteric forms such as 1,2-hydroxyamino, envisioning these small boronolectins as components in systems for drug release with tracking capability.
Asunto(s)
Antineoplásicos/farmacología , Benzamidas/química , Compuestos de Boro/farmacología , Doxorrubicina/farmacología , Nucleósidos/química , Nucleósidos/metabolismo , Antibióticos Antineoplásicos/farmacología , Antineoplásicos/química , Compuestos de Boro/química , Proliferación Celular , Doxorrubicina/química , Liberación de Fármacos , Colorantes Fluorescentes/química , Células HeLa , Humanos , Monosacáridos/químicaRESUMEN
A novel fluorescent molecular probe is reported, which is able to detect glycoproteins, especially mucins, with high sensitivity and with a turn-on response along with a large Stokes shift (>130â nm), within the biologically active window. The probe contains an aminotricarbocyanine as the fluorescent reporter with a linked benzoboroxole as the recognition unit, which operates through a dynamic covalent reaction between the boronic hemiester residue of the receptor and cis-diols of the analyte. The superior selectivity of the probe is displayed by the labeling of mucins present in Calu-3 cells. The new benzoboroxole fluorescent derivative gathers together key properties to make it a highly rated molecular probe: specificity, excellent solubility in water, and off-on near infrared emission. This probe is expected to be an excellent tool for imaging intracellular mucin to evaluate mucus-related diseases as well as a sensing strategy towards glycosylated structures with a high potential for theranostics approaches in biological samples.
Asunto(s)
Colorantes Fluorescentes/química , Glicoproteínas/análisis , Mucinas/análisis , Espectroscopía Infrarroja Corta/métodos , Compuestos de Boro/química , Células Epiteliales/efectos de los fármacos , Humanos , Estructura MolecularRESUMEN
Two fluorescent diboronic acid compounds (6a and 6b) with a dipeptide linker were synthesized as potential sensors for cell surface saccharide Lewis X (Le(X)). Compound 6a with a dipeptide (H-Asp-Ala-) as the linker was found to selectively label CHOFUT4 cells, which express Le(x), at micromolar concentrations, while non-Le(x)-expressing control cells were not labeled.
Asunto(s)
Antracenos/química , Ácidos Borónicos/química , Colorantes Fluorescentes/química , Oligosacáridos/metabolismo , Animales , Antracenos/síntesis química , Antracenos/metabolismo , Ácidos Borónicos/síntesis química , Ácidos Borónicos/metabolismo , Células CHO , Cricetinae , Cricetulus , Dipéptidos/química , Colorantes Fluorescentes/síntesis química , Colorantes Fluorescentes/metabolismo , Fucosiltransferasas/química , Fucosiltransferasas/metabolismo , Antígenos del Grupo Sanguíneo de Lewis , Microscopía Fluorescente , Oligosacáridos/genéticaRESUMEN
Carbohydrate-based biomarkers such as sialyl Lewis X are known to correlate with cancer formation and progression. By targeting sialyl Lewis X, we have developed a boronolectin-fluorophore conjugate, which was able to selectively label and image xenograft (sc) tumor. This represents the very first example that a small molecule capable of recognizing a carbohydrate biomarker was used for optical imaging application.
Asunto(s)
Compuestos de Boro/química , Colorantes Fluorescentes/química , Antígeno Lewis X/química , Monosacáridos/química , Imagen Óptica/métodos , Animales , Compuestos de Boro/síntesis química , Colorantes Fluorescentes/síntesis química , Células Hep G2 , Xenoinjertos , Humanos , Antígeno Lewis X/análisis , Antígeno Lewis X/metabolismo , Ratones , Monosacáridos/síntesis química , Antígeno Sialil Lewis XRESUMEN
There is great interest in developing boronolectins that are synthetic lectin mimics containing a boronic acid functional group for reversible recognition of diol-containing molecules, such as glycans and ribonucleotides. However, it remains a significant challenge to gain specificity. Here, we present a genetically encoded boronolectin which is a hybrid protein consisting of a noncanonical amino acid (ncAA) p-boronophenylalanine (pBoF), natural-lectin-derived peptide sequences, and a circularly permuted red fluorescent protein (cpRFP). The genetic encodability permitted a straightforward protein engineering process to derive a red fluorescent biosensor that can specifically bind uridine diphosphate N-acetylglucosamine (UDP-GlcNAc), an important nucleotide sugar involved in metabolic sensing and cell signaling. We further characterized the resultant boronic acid- and peptide-assisted UDP-GlcNAc sensor (bapaUGAc) both in vitro and in live mammalian cells. Because UDP-GlcNAc in the endoplasmic reticulum (ER) and Golgi apparatus plays essential roles in glycosylating biomolecules in the secretory pathway, we genetically expressed bapaUGAc in the ER and Golgi and validated the sensor for its responses to metabolic disruption and pharmacological inhibition. In addition, we combined bapaUGAc with UGAcS, a recently reported green fluorescent UDP-GlcNAc sensor based on an alternative sensing mechanism, to monitor UDP-GlcNAc level changes in the ER and cytosol simultaneously. We expect our work to facilitate the future development of specific boronolectins for carbohydrates. In addition, this newly developed genetically encoded bapaUGAc sensor will be a valuable tool for studying UDP-GlcNAc and glycobiology.
Asunto(s)
Monosacáridos , Uridina Difosfato , Animales , Uridina Difosfato/metabolismo , Monosacáridos/metabolismo , Aparato de Golgi/metabolismo , Ácidos Borónicos , Mamíferos/metabolismoRESUMEN
Bacterial infections, particularly by Gram-negative pathogens, have become a serious threat to global healthcare due to the diminishing effectiveness of existing antibiotics. We report a nontraditional therapy to combine three components in one macromolecular system, in which boronic acid adheres to peptidoglycan or lipopolysaccharide via boron-polyol based boronolectin chemistry, cationic metal polymer frameworks interact with negatively charged cell membranes, and ß-lactam antibiotics are reinstated with enhanced vitality to attack bacteria via evading the detrimental enzyme-catalyzed hydrolysis. These macromolecular systems exhibited high efficacy in combating pathogenic bacteria, especially Gram-negative strains, due to synergistic effects of multicomponents on interactions with bacterial cells. In vitro and in vivo cytotoxicity and hemolysis evaluation indicated that these multifunctional copolymers did not induce cell death by apoptosis, as well as did not alter the phenotypes of immune cells and did not show observable toxic effect on red blood cells.