Design, Synthesis, and Biological Application of Novel Photoaffinity Probes of Dihydropyridine Derivatives, BAY R3401

Abstract

To explore the molecular mechanisms of BAY R3401, four types of novel photoaffinity probes bearing different secondary tags were synthesized. Their potency for glycogenolysis was evaluated in primary human liver HL-7702 cells and HepG2 cells. Probe 2d showed the best activity in primary human liver HL-7702 cells and HepG2 cells, with IC50 values of 4.45 μM and 28.49 μM, respectively. Likewise, probe 5d showed IC50 values of 6.46 μM in primary human liver HL-7702 cells and 15.29 μM in HepG2 cells, respectively. Photoaffinity labeling experiments were also performed and protein bands larger than 170 kDa were specifically tagged by probe 2d. The results suggest that the synthesized probe 2d might be a very promising tool for the isolation of the target proteins of BAY R3401.

Design, Synthesis, and Use of Novel Photoaffinity Probes in Measuring the Serum Concentration of Glycogen Phosphorylase

“…Click chemistry was handled as shown in Scheme 2 . The alkyne 16 bearing a benzophenone photophore and a fluorescent tag was readily prepared in five steps starting from Lys(Boc)-OMe ( 14 ) and 4-benzoylbenzoic acid ( 15 ) [ 15 ]. The corresponding azide intermediate 13 and alkyne 16 were dissolved in CH 2 Cl 2 -H 2 O, followed by the addition of catalytic sodium ascorbate and CuSO 4 ·5H 2 O, resulting in triazole probe 1 in 11% yield.…”

“…The synthesis of the final probe 2 is shown in Scheme 3 . It took seven steps to prepare the dual-labeled alkyne 18 starting from N -Cbz- N ′-Boc- l -lysine 17 [ 15 ]. It was similar to probe 1 , where the reaction of the aforementioned azide 13 with the dual-labeled alkyne 18 resulted in probe 2 in 12% yield.…”

“…The tag-free probe 3 was also prepared as shown in Scheme 4 . According to the reported method, synthesis of the corresponding chloroacetyl compound 19 was based on Lys(Boc)-OMe ( 14 ) as well as 4-benzoylbenzoic acid ( 15 ) [ 15 ]. Then, probe 3 was synthesized by the CuI-catalyzed Huisgen (2 + 3) dipolar cycloaddition reaction (click chemistry), followed by a nucleophilic substitution reaction with sodium azide.…”

Enhanced optical imaging and fluorescent labeling for visualizing drug molecules within living organisms

“…However, most targets identified for fluorescence derivatives have been uncovered through blind screening, resulting in a low success rate and unexpected outcomes. To enhance signal-to-noise ratios and enable dynamic imaging, reversible chemical selection and catalytic processes with signal amplification offer promising solutions, in contrast to the prevailing irreversible chemical connections 165 , 166 , 167 . Nonetheless, challenges such as off-target effects and false positives persist in the clinical application of fluorescent drugs.…”

Glycogen phosphorylase inhibition alongside taxol chemotherapy synergistically elicits ferroptotic cell death in clear cell ovarian and kidney cancers

“…We also validated that this synergistic effect was specific to PYG by adopting a second PYG inhibitor, targeting a different site (Figure S1). We used BAY-R3401, which targets the AMP binding site of glycogen phosphorylase (42). We noted that BAY-R3401 similarly elicited a sharp drop in the doseresponse curves of all four CCC lines (Figure S1A), eliciting a 20-fold reduction in GI50 for OVTOKO and KOC7C cells, a 10-fold reduction for 786-O cells, and a 5-fold reduction for A498 cells; a trend also reflected in the CI values for the combination ( Figure S1B).…”