RESUMO
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.
Assuntos
Antracenos/química , Ácidos Borônicos/química , Corantes Fluorescentes/química , Oligossacarídeos/metabolismo , Animais , Antracenos/síntese química , Antracenos/metabolismo , Ácidos Borônicos/síntese química , Ácidos Borônicos/metabolismo , Células CHO , Cricetinae , Cricetulus , Dipeptídeos/química , Corantes Fluorescentes/síntese química , Corantes Fluorescentes/metabolismo , Fucosiltransferases/química , Fucosiltransferases/metabolismo , Antígenos do Grupo Sanguíneo de Lewis , Microscopia de Fluorescência , Oligossacarídeos/genéticaRESUMO
Carbohydrate antigens with subterminal fucosylation have been implicated in the development and progression of several cancers, including hepatocellular carcinoma (HCC). Fluorescent sensors targeting fucosylated carbohydrate antigens could potentially be used for diagnostic and other applications. We have designed and synthesized a series of 26 diboronic acid compounds as potential fluorescent sensors for such carbohydrates. Among these compounds, 7q was able to fluorescently label cells expressing high levels of sLex (HEPG2) within a concentration range of 0.5 to 10 microM. This compound (7q) did not label cells expressing Lewis Y (HEP3B), nor cells without fucosylated antigens (COS7). This represents the first example of a fluorescent compound labeling cells based on cell surface carbohydrate structures.
Assuntos
Ácidos Borônicos/química , Carcinoma Hepatocelular/metabolismo , Neoplasias Hepáticas/metabolismo , Oligossacarídeos/análise , Oligossacarídeos/genética , Animais , Ácidos Borônicos/síntese química , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/imunologia , Linhagem Celular , Fluorescência , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/imunologia , Estrutura Molecular , Oligossacarídeos/química , Antígeno Sialil Lewis XRESUMO
A glucose selective diphenylboronic acid fluorescent sensor (10a) with a K(a) of 1472M(-1) has been synthesized and evaluated. This sensor shows a 43- and 49-fold selectivity for glucose over fructose and galactose, respectively. The binding affinity improvement is about 300-fold and the selectivity improvement for glucose over fructose is about 1400-fold compared with the monoboronic acid compound, phenylboronic acid. 1H NMR studies indicate that sensor 10a binds with alpha-D-glucofuranose in a bidentate manner (1:1 ratio).
Assuntos
Técnicas Biossensoriais/métodos , Compostos de Boro/química , Corantes Fluorescentes/química , Glucose/análise , Compostos de Boro/síntese química , Fluorescência , Corantes Fluorescentes/síntese química , Glucose/análogos & derivados , Espectroscopia de Ressonância Magnética , Sensibilidade e Especificidade , Espectrometria de FluorescênciaRESUMO
A series of fluorescent diboronic acids was synthesized in nine steps as potential sensors for sialyl Lewis X (sLex). The fluorescent binding studies of these sensors with sLex were carried out in a mixed aqueous solution. Compound 7e was found to show the strongest fluorescence enhancement upon binding with sLex. Using cell cultures, 7e was shown to label sLex-expressing HEPG2 cells at 1 microM, while non-sLex-expressing cells were not labeled.
