With H2S and 1 BODIPY-flavonoid nanoreagent with O2-synergistic CO release capability and its preparation and application

Abstract

The present invention belongs to the field of biochemistry and relates to a method for preparing a biochemical product having H2S and 1 O2 synergistically drives the BODIPY flavonoid nanoreagent of CO release ability and its preparation method and application. It is wrapped in the hydrophobic inner cavity of the micelle formed by mPEG DSPE2000, and the outer layer of the micelle is a silicon shell formed by N trimethoxysilylpropyl N, N, N tri-n-butylammonium bromide, which significantly improves the water solubility of the molecule. The core-shell structure of the formed nanomolecule is relatively stable, which can significantly improve the stability of the molecule in water. The iodinated BODIPY flavonoid compound of the present invention has very good photodynamic effect and CO release ability, has a very high singlet oxygen production efficiency under 660nm light irradiation, and has excellent stability. It has and its considerable application prospects in photodynamic and gas therapy in the fields of biochemistry and medicine. The phototherapy reagent synthesis method provided by the present invention is simple and relatively low in cost.

Description

Technical Field

[0001] The present invention belongs to the field of biochemistry and relates to a tumor treatment agent, its synthesis, and application, which can be used for the synergistic treatment of tumors with PDT and CO gas. Specifically, it relates to a method for synthesizing organic fluorescent molecules and the application of biophotodynamic synergistic gas therapy. Background Art

[0002] Photodynamic therapy (PDT) is an effective, noninvasive treatment for various cancers. Compared to conventional chemotherapy, PDT offers inherent advantages such as minimal invasiveness, ease of use, short treatment time, rapid recovery, and high spatial selectivity, making it a promising anti-tumor treatment with unlimited potential. PDT primarily utilizes photosensitizers as light-sensitive drugs, which are activated by light irradiation to produce reactive oxygen species (ROS) with lethal potential, disrupting cell function and killing cells in the exposed area. Gas therapy, as an emerging field, is attracting increasing attention. Gas therapy has many advantages, including minimal side effects on normal tissues and greater safety; and the absence of other byproducts, making it an emerging "green" treatment. A growing number of studies have shown that CO, a typical gas therapy molecule, has a strong anti-tumor effect.

[0003] Compared with a single treatment method, the combination of multiple drugs or treatment methods can effectively improve the treatment effect and prolong the patient's life. Therefore, it is challenging and practical to construct a low-cost material with combined treatment capabilities. Based on this background, the present invention designed and synthesized a NanoBDP2I-S-HF that can identify and consume H2S, produce PDT and CO. It has the ability to consume H2S while identifying H2S, and is used for photodynamic and gas combined treatment of tumors. NanoBDP2I-S-HF consists of two parts. The first part is iodine-substituted Bodipy, which can efficiently produce 1 O2; the second part is a flavonoid derivative, which can be 1 O2 is split to release CO. The two parts are connected by a disulfide bond that is specifically cleaved by hydrogen sulfide. Only in HCT116 cells that overexpress H2S can the disulfide bond be broken to release the precursor compound 3-hydroxyflavone that can release CO. The core-shell structure of the nanomolecule formed after encapsulation is relatively stable, significantly improving the stability of the molecule in water and increasing the molecule's response rate to hydrogen sulfide. Compared with single treatment modes, the NanoBDP2I-S-HF nanoreagent with combined treatment capabilities has shown superior anti-tumor effects both in vitro and in vivo. Summary of the Invention

[0004] The first object of the present invention is to provide a method for the first time having H2S and 1BODIPY-flavonoid nanoreagent with O2 synergistically driven CO release capability and preparation method thereof;

[0005] The second object of the present invention is to provide a method for the first time having H2S and 1 BODIPY-flavonoid phototherapy agent with O2-synergistic CO release capability and preparation method thereof;

[0006] The third purpose of the present invention is to provide for the first time a nanoreagent with PDT effect that can release CO after being activated by hydrogen sulfide for use in anti-tumor applications in vivo and in vitro.

[0007] The present invention is achieved through the following technical solutions:

[0008] A kind of H2S and 1 BODIPY-flavonoid phototherapy agents with O2-synergistic CO release capability, including BDP2I-S-HF, BDP2I-S-OH, and BDP2I-C-OH;

[0009] The structural formula of the BDP2I-S-HF compound is shown below:

[0010]

[0011] The structural formula of the BDP2I-C-OH is:

[0012]

[0013] The structural formula of the BDP2I-S-OH is:

[0014]

[0015] The present invention provides for the first time a nano anti-tumor agent (i.e., a PDT agent with H2S specific activation and CO gas synergistic therapy) 1 O2 synergistically drives CO release capability of BODIPY-flavonoid nanoreagent), the structure of which is shown in Formula I.

[0016]

[0017] The present invention also provides a method having H2S and 1The invention discloses a BODIPY-flavonoid nanoreagent with the ability to synergistically drive CO release by O2, and is prepared by the following steps: mixing mPEG-DSPE-2000 and a dilute hydrochloric acid solution and sonicating until clear and transparent; adding BDP2I-S-HF, BDP2I-C-OH or BDP2I-S-OH according to claim 1 dissolved in DMSO, and then adding TBNBr; and finally dialyzing and filtering to obtain nanoparticles NanoBDP2I-S-HF, NanoBDP2I-C-OH or NanoBDP2I-S-OH.

[0018] The present invention has H2S and 1 The BODIPY-flavonoid nanoreagent has the ability to synergistically drive CO release by O2. The BODIPY-flavonoid nanoreagent uses 660nm near-infrared light as a light source to irradiate the phototherapy reagent so that it can efficiently produce cytotoxic singlet oxygen. The singlet oxygen can spontaneously oxidize the 3-HF derivative released after hydrogen sulfide activation to release CO. The phototherapy reagent is activated by hydrogen sulfide while being consumed by hydrogen sulfide, which can inhibit the migration of tumor cells, thereby improving the anti-tumor effect of the phototherapy reagent.

[0019] The present invention provides the above-mentioned bodipy-flavonoid derivative nano anti-tumor agent, which has strong absorption near 660nm and efficiently produces 1 O2, and used it for H2S consumption, photodynamic therapy and CO gas combined therapy on HCT116 cells and tumor mouse models.

[0020] The H2S and 1 The BODIPY-flavonoid nanoreagent has the ability to synergistically drive CO release by O2. The BODIPY-flavonoid nanoreagent uses 660nm near-infrared light as a light source to irradiate the phototherapy agent to efficiently produce cytotoxic singlet oxygen. The singlet oxygen can spontaneously oxidize the 3-HF derivative released after hydrogen sulfide activation to release CO. The phototherapy agent is activated by hydrogen sulfide while being consumed by hydrogen sulfide, which can inhibit the migration of tumor cells, thereby improving the anti-tumor effect of the phototherapy agent. As a large amount of 1 The phototherapy agent exhibits remarkable anti-tumor effects by releasing O2 and CO, as well as consuming hydrogen sulfide. This CO release, triggered by an intracellular cascade, occurs only in the illuminated area, effectively avoiding CO-related side effects.

[0021] The present invention has H2S and 1 The preparation routes of BDP2I-S-HF phototherapy reagent, BDP2I-C-OH and BDP2I-S-OH with the ability of O2 to synergistically drive CO release are as follows:

[0022]

[0023] The present invention also provides a method having H2S and 1 Application of BODIPY-flavonoid nanoreagents with O2-synergistically driven CO release ability in anti-tumor studies in vivo and in vitro.

[0024] The present invention also provides a method having H2S and 1 Application of BODIPY-flavonoid nanoreagents with O2-synergistically driven CO release ability in the preparation of anti-tumor drugs.

[0025] The present invention provides a method having H2S and 1 This BODIPY-flavonoid nanoreagent, synergistically driven by O2, is encapsulated within the hydrophobic inner cavity of micelles formed from mPEG-DSPE2000. The outer layer of the micelle is a silica shell formed from N-trimethoxysilylpropyl-N,N,N-tri-n-butylammonium bromide, significantly improving the molecule's water solubility. The resulting nanomolecule has a relatively stable core-shell structure, significantly improving its stability in water. The iodinated BODIPY-flavonoid compound exhibits excellent photodynamic effect and CO2 release, high singlet oxygen generation efficiency under 660nm light irradiation, and excellent stability. It holds great promise for applications in photodynamic and gas therapy in biochemistry and medicine.

[0026] Beneficial technical effects of the present invention:

[0027] The nano-agent with combined tumor treatment capability provided by the present invention has a simple synthesis method, relatively low cost, and excellent in vitro and in vivo anti-tumor effects. Compared with single treatment methods, the anti-tumor effect of this combined treatment agent is significantly improved, with the highest tumor inhibition rate. (See attached Figure 7 ), for the first time, the application of PDT-assisted hydrogen sulfide-activated CO-releasing nanoreagents in anti-tumor applications.

[0028] The organosilicon nanoagent provided by the present invention has excellent optical properties. It absorbs in the near-infrared region, where near-infrared light has strong tissue penetration and minimal phototoxicity. Irradiation with near-infrared light (660 nm) for 10 minutes produces highly efficient singlet oxygen. Hydrogen sulfide specifically breaks disulfide bonds, releasing flavonoid derivatives that can be cleaved by singlet oxygen to release CO. This nanoagent exhibits excellent anti-tumor properties both in vitro and in vivo, and has promising applications in the biomedical field. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 The synthetic route of BDP2I-S-HF, BDP2I-S-OH and BDP2I-C-OH is shown in FIG.

[0030] Figure 2 The BDP2I-S-HF in CDCl3 shown in formula I 1 H NMR spectrum.

[0031] Figure 3 The UV absorption spectrum of BDP2I-S-HF shown in Formula I in DMSO when the biological concentration is 2 μmol / L.

[0032] Figure 4 BDP2I-S-HF is represented by formula I, with DPBF as 1 UV absorption spectra of O2 indicator at different times of 660nm laser irradiation (Note: power density is 10mWcm -2 ).

[0033] Figure 5 Cell scratch test of BDP2I-S-HF nanoparticles.

[0034] Figure 6 In order to use 2′,7′-dihydrodichlorofluorescein diacetate (DCFH-DA) as an intracellular reactive oxygen species indicator, confocal imaging of the reactive oxygen species produced in cells by the BDP2I-S-HF nanoreagent represented by formula I was performed.

[0035] Figure 7 The phototoxicity and dark toxicity of NanoBDP2I-C-OH, NanoBDP2I-S-OH, and NanoBDP2I-S-HF nanoreagents at different concentrations on HCT116 cells.

[0036] Figure 8 This is a diagram showing the effects of NanoBDP2I-C-OH, NanoBDP2I-S-OH, and NanoBDP2I-S-HF in one cycle of animal treatment. DETAILED DESCRIPTION

[0037] The present invention will be further described below with reference to the following examples. Those skilled in the art should understand that the examples are only for illustration and do not constitute any limitation to the present invention.

[0038] Example 1

[0039] Synthesis of three compounds

[0040]

[0041] The specific method is:

[0042] Compound 1 was synthesized according to the method reported in Y Zhou, Y Cheung, C Ma, etc. J. Med. Chem. 2018, 61, 3952-3961.

[0043] Synthesis of compound 2

[0044] Compound 1 (500 mg, 1.17 mmol) and iodine (447 mg, 1.76 mmol) were dissolved in 100 mL of ethanol. HIO₃ (413 mg, 2.34 mmol) was then dissolved in 1 mL of deionized water and added dropwise to the reaction system. The temperature was slowly raised to 60°C with stirring and monitored by TLC. After completion, the reaction was quenched by adding a saturated NaS₂O₃ solution. The product was extracted with CHCl₂ and dried over anhydrous Na₂SO₄. After concentration, the crude product was separated by column chromatography using a developing solvent ratio of PE:DCM = 2:1. The yield was 85%. 1 H NMR (400MHz, CDCl3) δ = 7.09 (d, 2H), 6.98 (d, 2H), 4.64 (s, 2H), 4.23 (q, 2H), 2.57 (s, 6H), 1.36 (s, 6H), 1.25 (t, 3H).

[0045] Synthesis of compound 3

[0046] Dissolve p-anisaldehyde (160 mg, 1.19 mmol) and compound 2 (400 mg, 0.590 mmol) in 25 mL of toluene, add piperidine (0.2 mL) and acetic acid (0.2 mL). Reflux the reaction system at 115°C overnight. After completion, dilute the reaction with DCM, combine the organic phases, wash with deionized water, dry over anhydrous sodium sulfate, and remove the solvent in vacuo. The resulting crude product is separated by column chromatography using a 1:1 ratio of PE to DCM. The yield is 25%. 1 H NMR(400MHz, CDCl3)δ=8.07(d,2H),7.50-7.55(m,6H),7.15(d,2H),7.00(d,2H) ,6.89(d,4H),4.65(s,2H),4.24(q,2H),3.80(s,6H),1.42(s,6H),1.26(t,3H).

[0047] Synthesis of compound 4

[0048] Compound 3 (300 mg, 0.33 mmol) and potassium carbonate (317 mg, 2.29 mmol) were dissolved in a 1:1 THF:H₂O mixture (25 mL, v / v). The reaction system was stirred at 50°C for 24 hours. After completion, the reaction was cooled to room temperature and the pH was adjusted to 7 with 1M hydrochloric acid. The system was diluted with DCM, and the combined organic phases were washed with water, dried over anhydrous sodium sulfate, and the solvent removed in vacuo. The resulting compound 4 was used directly in the next reaction without purification.

[0049] Synthesis of compound BDP2I-S-OH

[0050] Compound 4 (200 mg, 0.23 mmol), 2-hydroxyethyl disulfide (109 mg, 0.7 mmol), EDCI (89 mg, 0.58 mmol), and DMAP (4 mg, 0.03 mmol) were dissolved in 30 mL of DCM and stirred overnight at room temperature under argon. After completion of the reaction, the system was diluted with DCM, and the combined organic phases were washed with water, dried over anhydrous sodium sulfate, and the solvent removed in vacuo. The resulting crude product was separated by column chromatography using a developing solvent ratio of PE:DCM = 1:2. The yield was 45%. 1 H NMR(400MHz, DMSO-d6)δ=8.08(d,2H),7.59(d,4H),7.43(d,2H),7.36(d,2H),7.17(d,2H),7.07(d,4H),4.9 4(s,2H),4.92(t,1H),4.41(t,2H),3.83(s,6H),3.64(dd,6.4,2H),3.00(t,2H),2.83(t,2H),1.47(s,6H).

[0051] Synthesis of compound BDP2I-C-OH

[0052] Compound 4 (200 mg, 0.23 mmol), 1,6-hexanediol (82.6 mg, 0.7 mmol), EDCI (89 mg, 0.58 mmol), and DMAP (4 mg, 0.03 mmol) were dissolved in 30 mL of DCM and stirred overnight at room temperature under argon. After completion of the reaction, the system was diluted with DCM, and the combined organic phases were washed with water, dried over anhydrous sodium sulfate, and the solvent removed in vacuo. The crude product was separated by column chromatography using a developing solvent ratio of PE:DCM = 1:2. The yield was 45%. 1 H NMR (400MHz, DMSO-d6)δ=8.08(d,2H),7.59(d,4H),7.43(d,2H),7.36(d,2H),7.16(d,2H),7.07(d,4H),4.93(s,2H),4. 36(t,1H),4.14(t,2H),3.82(s,6H),3.40(t,2H),1.63-1.54(m,2H),1.45(d,6H),1.45-1.37(m,2H),1.35-1.28(m,4H).

[0053] Compound 5 was synthesized according to the method reported in the literature. S Kumar, R Ambatwar, V Gupta, etc. Research on Chemical Intermediates (2023) 49: 901-915.

[0054] Synthesis of compound 6

[0055] 3-Hydroxyflavone (200 mg, 0.84 mmol), 4-nitrophenyl chloroformate (252 mg, 1.26 mmol), and DMAP (154 mg, 1.26 mmol) were thoroughly dissolved in DCM (10 mL) and stirred under argon at 25°C for 12 h. The reaction was monitored by thin-layer chromatography. After completion, the reaction mixture was diluted with DCM, washed three times with water, dried over Na2SO4, and the solvent removed in vacuo. The product was used directly in the next step without purification.

[0056] Synthesis of compound BDP2I-S-HF

[0057] Compound 6 (50 mg, 0.05 mmol), compound BDP2I-S-OH (30 mg, 0.07 mmol), and DMAP (8.6 mg, 0.07 mmol) were dissolved in 10 mL of DCM and stirred overnight at room temperature under argon. After completion of the reaction, the system was diluted with DCM, and the combined organic phases were washed with water, dried over anhydrous sodium sulfate, and the solvent removed in vacuo. The resulting crude product was separated by column chromatography using a developing solvent ratio of PE:DCM = 1:2. The yield was 30%. 1 H NMR (400MHz, CDCl3) δ=8.27(dd,1H),8.14(d,2H),7.96-7.91(m,2H),7.75-7.69(m,1H),7.66-7.60(m,5H),7.60-7.53(m,5H),7. 45(t,1H),7.17(d,2H),7.08(d,2H),6.98(d,4H),4.78(s,2H),4.55(dt,4H),3.88(s,6H),3.07(t,2H),3.03(t,2H),1.48(s,6H). 13C NMR(600MHz, DMSO)δ=171.57,168.87,161.14,158.98,156.32,155.56,152.16,150.1 9,145.99,139.04,135.34,133.56,133.22,132.26,130.12,129.49,129.38,129.06,1 28.70,127.43,126.30,125.51,123.21,119.21,116.50,116.00,115.27,84.67,67.38,65.18,62.79,55.87,36.95,36.61,31.62,30.29,29.49,29.19,22.56,17.71,14.42.

[0058] Preparation of nanoprobes

[0059] mPEG-DSPE-2000 (20 mg) was weighed into a 5 mL clear glass vial. 2 mL of dilute hydrochloric acid solution was added and sonicated until clear. Then, 100 μL of DMSO-dissolved small molecules BDP2I-C-OH (0.2 mg), BDP2I-S-OH (0.2 mg), and BDP2I-S-HF (0.2 mg) were quickly added, and sonication continued for 20 minutes. Subsequently, 100 μL of TBNBr was added while vigorously stirring with a magnetic stir bar, and stirring continued at room temperature for 6 hours. The solution was then transferred to a dialysis membrane and dialyzed against deionized water for 6 hours to remove free dye and organic solvent. After dialysis, the sample was removed and filtered through a 0.22 μm polyvinylidene fluoride (PVDF) filter to obtain nanoparticles: NanoBDP2I-C-OH, NanoBDP2I-S-OH, and NanoBDP2I-S-HF.

[0060] Example 2

[0061] HCT116 cells were cultured in Dulbecco's Eagle medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin at 37°C in a humidified atmosphere of 5 / 95 CO2 / air incubator for 24 h. Cells were seeded in glass-bottomed culture dishes and allowed to adhere for 24 h before the experiment and washed with PBS buffer (pH 7.4) before imaging. The probe concentrations for cytotoxicity experiments were 0, 2, 4, 6, and 8 μM, and phototoxicity was determined using a 660 nm LED light source (10 min, 0.1 W / cm 2 ),like Figure 7As shown in the figure, the intracellular reactive oxygen species test experiment was performed using a Leica TCS SP8 confocal microscope and a 10x lens, with an excitation wavelength of 488 nm and a collection wavelength of 490-540 nm. Figure 6 shown.

[0062] All in vivo experiments in this invention complied with the regulations and rules for the breeding and use of experimental animals. The tumor-bearing mice used in the experiments were purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd. and were kept in sterile cages in a laminar flow hood in a sterile room and fed with water and food treated with high-pressure steam.

[0063] NanoBDP2I-S-OH, NanoBDP2I-C-OH, and NanoBDP2I-S-HF nanoparticles were subcutaneously injected into the tumor site of tumor-bearing mice at a dose of 25 nmol. After injection, laser irradiation (Note: laser wavelength: 660 nm, power density: 0.1 W cm -2 ), and recorded the size of the mouse tumors every day. It was found that NanoBDP2I-S-HF, which has the ability to combine treatments, has the best anti-tumor effect. This shows that our therapeutic agent has excellent H2S consumption, photodynamic and CO generation capabilities, and can be successfully used in combined tumor treatment. See Appendix Figure 8 . Figure 8 The effect of NanoBDP2I-C-OH, NanoBDP2I-S-OH, and NanoBDP2I-S-HF on animals after one cycle of treatment (Note: the dosage is 25 nmol, the laser wavelength is 660 nm, and the power density is 0.08 W cm -2 ), A, B, C, D, E, and F represent the no-drug and no-light group, the light-only group, the NanoBDP2I-S-HF-only group, the NanoBDP2I-C-OH and light-only group, the NanoBDP2I-S-OH and light-only group, and the NanoBDP2I-S-HF and light-only group, respectively. This indicates that combined therapy indeed exhibits superior anti-tumor effects compared to either single therapy.

[0064] Example 3

[0065] HCT116 cells in 6-well plates were treated with NanoBDP2I-S-HF (20 μM) for 2 hours and compared to untreated cells. A line was drawn smoothly with a toothpick tip, and the cells were then incubated with FBS-free medium for 24 hours. Images of the scratch at 0 and 24 hours were observed using an inverted microscope, and cell migration patterns were compared and recorded (e.g., Figure 5 shown). Figure 5The cell scratch experiment of BDP2I-S-HF nanoparticles shows that the nanoparticles can not only react with hydrogen sulfide, but also consume hydrogen sulfide in cells, inhibit cell migration and proliferation, and thus induce cancer cell death.

[0066] Example 4

[0067] HCT116 cells were seeded in confocal microscopy and adhered to the wall after 12 hours. The experiment was divided into 4 groups: (1) cells were not treated and DCFH-DA (10 μM) was added and cultured for 30 min; (2) cells were illuminated by 660 nm LED (100 mW cm -2 ) After irradiation with light for 20 min, DCFH-DA (10 μM) was added and cultured for 30 min; (3) the cells were incubated with NanoBDP2I-S-HF (10 μM) in the dark and DCFH-DA (10 μM) was added and cultured for 30 min; (4) the cells were incubated with NanoBDP2I-S-HF (10 μM) for 2 hours, DCFH-DA (10 μM) was added and cultured for 30 min, washed with PBS buffer and illuminated with 660 nm LED light (100 mW cm -2 ) for 20 min. Finally, the fluorescence microscope Carl Zeiss LSM710 was used to evaluate 1 The excitation wavelength is 488nm, and the emission band is collected at 490-540nm (such as Figure 6 shown). Figure 6 In order to use 2′,7′-dihydrodichlorofluorescein diacetate (DCFH-DA) as an intracellular reactive oxygen species indicator, confocal imaging of the reactive oxygen species produced by the BDP2I-S-HF nanoreagent represented by Formula I in cells was performed (Note: laser wavelength: 660 nm, power density: 0.1 W cm -2 The excitation wavelength of confocal imaging was 488 nm, and the emission wavelength collection range was 490-540 nm. ) This indicates that the phototherapy agent has good 1 O2 production capacity.

[0068] Figure 4 BDP2I-S-HF is represented by formula I, with DPBF as 1 UV absorption spectra of O2 indicator at different times of 660nm laser irradiation (Note: power density is 10mWcm -2 ). This indicates that the phototherapy agent has a high 1 O2 generation capacity, 1 The O2 yield was 67.9%.

[0069] Figure 7The phototoxicity and dark toxicity of NanoBDP2I-C-OH, NanoBDP2I-S-OH, and NanoBDP2I-S-HF nanoreagents at different concentrations on HCT116 cells (Note: excitation wavelength: 660 nm, power density: 0.1 W cm -2 ), where A, B, and C represent the dark toxicity of NanoBDP2I-C-OH, NanoBDP2I-S-OH, and NanoBDP2I-S-HF, respectively; D, E, and F represent the phototoxicity of NanoBDP2I-C-OH, NanoBDP2I-S-OH, and NanoBDP2I-S-HF, respectively. These results demonstrate that the combined therapy exhibits superior cancer cell killing compared to either therapy alone.

Claims

1. With H2S and 1 O2 synergistically drives CO release capability of BODIPY-flavonoid phototherapy agent, characterized in that, They are BDP2I-S-HF, BDP2I-S-OH and BDP2I-C-OH; The structural formula of the BDP2I-S-HF compound is shown below: The structural formula of the BDP2I-C-OH is: The structural formula of the BDP2I-S-OH is:

2. A method according to claim 1 having H2S and 1 The preparation route of BODIPY-flavonoid phototherapy reagent with O2 synergistically driven CO release ability is as follows:

3. A kind of H2S and 1 O2 synergistically drives the CO release ability of BODIPY-flavonoid nanoreagent, characterized by: The method is prepared by the following steps: mixing mPEG-DSPE-2000 and a dilute hydrochloric acid solution and ultrasonicating the mixture until the mixture becomes clear and transparent; adding BDP2I-S-HF, BDP2I-C-OH or BDP2I-S-OH according to claim 1 dissolved in DMSO and then adding TBNBr; and finally dialyzing and filtering the mixture to obtain nanoparticles NanoBDP2I-S-HF, NanoBDP2I-C-OH or NanoBDP2I-S-OH.

4. A method according to claim 3 having H2S and 1 Application of BODIPY-flavonoid nanoreagent with O2-synergistically driven CO release ability in in vitro anti-tumor research.

5. A method according to claim 3 having H2S and 1 Application of BODIPY-flavonoid nanoreagents with O2-synergistically driven CO release ability in the preparation of anti-tumor drugs.

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