Design, Synthesis and Application of CO Donor with Green Light Controlled Release and Fluorescence Enhancement Response

doi:10.6023/cjoc202407023

Abstract

Carbon monoxide (CO) is not only a toxic gas, but also an important gas signaling molecule that plays crucial regulatory roles in various physiological and pathological processes, which include neurotransmission, vasodilation, mitigation of inflammatory response, and anti-cancer activities. Therefore, development of CO donors for controlled release of CO with simutaneous fluorescence signal generation for quantification and tracking purpose is particularly useful for CO signal transduction and quantity-activity relationship studies. Herein, a novel photo-controlled CO donor BODIPY-Fla-1 was designed and synthesized using 3-hydroxyflavone (HF) as the photo-triggered CO-releasing moiety, which is conjugated to a boron-dipyrromethene (BODIPY) fluorophore at its 4' position of B ring via a double-bond linkage. It can achieve photo- triggered CO release under green light-emitting diode (LED) light irradiation and aerobic condition with simutaneous fluorescence turn-on responses at 562 nm for convenient tracking CO release via fluorescence imaging at green channel in live cell studies. It was also verified that the CO released by photolysis of BODIPY-Fla-1 can effectively reduce the level of intracellular superoxide radical and alleviate oxidative stress. This incorporation of a conjugated fluorophore moiety via a double bond linkage provides a new design strategy for development of HF-based CO donors with longer photolysis light and fluorescence response wavelength.

Key words: carbon monoxide, CO donor, photo-triggered release, oxiditive stress, green fluorescence

Cite this article

Wanjin Xing, Yirong Zhang, Junwei Chen, Yang Li, Huan Xu, Wei Wang, Kaiyan Lou. Design, Synthesis and Application of CO Donor with Green Light Controlled Release and Fluorescence Enhancement Response[J]. Chinese Journal of Organic Chemistry, 2025, 45(5): 1698-1706.

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Fig. & Tab. 9

Figure 1. (a) Reported 3-HF derivatives for photo-controlled CO donors; (b) Representative structures of 3-HF derivatives with photolysis wavelength at visible or near-infrared region; (c) This work: incorporation of a boron-dipyrromethene (BODIPY) fluorophore conjugated to 4' position of 3-hydroxyflavone (HF) via a double-bond linkage

Scheme 1. Synthesis route of probe BODIPY-Fla-1

Figure 2. (a) Time-dependent UV absorption spectra of BODIPY-Fla-1 (10 μmol/L) upon photolysis; (b) Time-dependent fluorescence emission spectrum of BODIPY-Fla-1 (10 μmol/L, λex=520 nm) upon photolysis All measurements were performed in 10 mmol/L pH=7.4 PBS buffer. Photolysis reactions were performed under a 15 W green LED light.

Figure 3. (a) Time-dependent fluorescence emission spectra of 10 μmol/L BODIPY-Fla-1 preirradiated with green LED light for 15 min incubated with NF-APC (10 μmol/L) and Na2PdCl4 (60 μmol/L) (λex=620 nm); (b) Time-dependent absorption spectra of the probe BODIPY-Fla-1 (50 μmol/L) coincubated with Hb (4.2 μmol/L) upon irradiated with green LED light; (c) Corresponding time-dependent calculated COHb concentration upon photoirradiation All measurements were performed in 10 mmol/L pH=7.4 PBS buffer, the light source is green LED, 15 W

Figure 4. Reaction scheme of CO release from photolysis of BODIPY-Fla-1

Figure 5. Time-dependent of fluorescent intensity changes of probe BODIPY-Fla-1 (10 μmol/L) at 562 nm (λex=520 nm) in PBS buffer (10 mmol/L, pH=7.4) after the periodic irradiation with green LED (15 W) in every 15 min

Figure 6. (a) Percentage of viable HeLa cells after treated with different concentrations of BODIPY-Fla-1 for 8 h; (b) Percentage of viable RAW264.7 cells after treated with different concentrations of BODIPY-Fla-1 for 8 h

Figure 7. Fluorescence imaging of CO-release with probe BODIPY-Fla-1 in living RAW 264.7 cells. (a) Cells were incubated in normal media; (b) Fluorescence image of cells incubated in 10 μmol/L probe BODIPY-Fla-1 for 10 min; (c) Fluorescence image of cells incubated in 10 μmol/L probe BODIPY-Fla-1 for 10 min followed by 30 min of irradiation; (d~f) Corresponding bright-field images; (g) Relative fluorescence intensities of fluorescence images in different cell groups (Fluorescence images were captured with λex=340~380 nm and λem>425 nm; irradiation light source: green LED,15 W. The error bar represents the standard deviation of three repetitions, ****P≤0.0001, ***P≤0.001)

Figure 8. Anti-oxidative stress effect of different concentrations of BODIPY-Fla-1 on A549 cells in dark or under green light irradiation Error bars represent standard deviations of three replicate images in the same group, ***P≤0.001, *P≤0.1, ns: no significant difference.

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