Synthesis of a BODIPY Library and Its Application to the Development of Live Cell Glucagon Imaging Probe

Abstract

The first BODIPY library (BD) was synthesized, and a highly selective glucagon sensor, Glucagon Yellow (BD-105), was discovered by fluorescence image-based screening method. BD library was synthesized via a Knoevenagel-type condensation reaction with 160 benzaldehydes and the 1,3 dimethyl-BODIPY scaffold. Using BD compounds, a fluorescence image-based screening was performed against three cell lines including AlphaTC1 and BetaTC6 cells which secret glucagon and insulin, respectively, and HeLa as control cells. Out of the 160 candidate probes, one compound, Glucagon Yellow, exhibited selective staining only in AlphaTC1 cells. The selectivity of Glucagon Yellow toward glucagon was confirmed in vitro by comparison of its fluorescence intensity change against 19 biologically relevant analytes. Subsequent immunostaining experiments revealed that Glucagon Yellow and the glucagon antibody colocalized in pancreas tissue, showing a high quantitative correlation analysis by the Pearson's coefficient constant (R(r) = 0.950). These results demonstrated the potential application of Glucagon Yellow as a glucagon imaging agent in live cells and tissues.

Fluorescent Dyes Used in Polymer Carriers as Imaging Agents in Anticancer Therapy

“…A BODIPY derivative labeling of recombinant protein molecules FABP7-expressing 293HEK cells (i.e., a protein encoded by a gene) are well-known as intracellular markers of radial glial cells [106,107]. Chang et al [108,109] discovered BODIPY which selectively stain Langerhans islets composed of glucagonsecreting α-cells (in the presence of glucagon the fluorescent compound was named Glucagon Yellow) and insulin-secreting β-cells in pancreas tissue sections. They also developed chemical imaging probes that specifically stain live microglia of brain cells [110].…”

Quantitative Structure‐Fluorescence Property Relationship Analysis of a Large BODIPY Library

“…The synthesis of 160 library members was reported previously by us. [35] The synthesis of an additional 160 BODIPY compounds will be published elsewhere. All compounds were purified by semi-preparative LC and characterized by HPLC/ MS, ensuring the high quality of our experimental data.…”

“…[35] Absorption and fluorescence emission spectra were recorded and the absorption (l abs ) and fluorescence emission (l em ) wavelength maxima were determined. 288 compounds with high quality spectra were chosen for QSPR analysis (BD 288 library) and their spectroscopic properties are summarized in Table 1 and Figure 2.…”

“…[35] BODIPY has outstanding photophysical properties as a fluorescent scaffold, such as high photostability, high fluorescent quantum yield, a high extinction coefficient, and narrow emission bandwidth. [36] BODIPY dyes are excitable with visible light and their chemical modification has been well studied.…”

Tuning the optical properties of BODIPY dye through Cu(I) catalyzed azide-alkyne cycloaddition (CuAAC) reaction

“…500 nm), narrow emission bands and high emission intensity. They have been used as laser dyes [9][10][11][12][13][14][15][16][17][18][19][20], markers [21,22], sensors [23][24][25][26][27][28] in modern chemistry and biology [29]. It is well known that the functionalities on 3-, 4-, 5-and 8-positions of the BODIPY core (Figure 1) strongly impact its optical properties [30][31][32][33], which led to many efforts to design BODIPY-based sensor molecules [34][35][36][37][38][39].…”