A reaction system and method for releasing aldehyde compounds from BODIPY photocage molecules induced by visible light

Abstract

The present invention provides a reaction system and method for releasing aldehyde compounds by visible light-induced BODIPY photocage molecules. The reaction system comprises: reactants: BODIPY photocage molecules containing diene groups; reaction medium; reaction conditions: aerobic environment and light source irradiation conditions; reactants formed by the reaction system include: aldehyde compounds and BODIPY-aldehyde compounds. The present invention can also construct a prodrug molecule with targeting properties by modifying the BODIPY photocage molecule with a targeting group, and use it for targeted and controlled release of aldehyde drugs in cells and organelles, and monitor it in real time through a biothiol fluorescent probe.

Description

Technical Field

[0001] The invention belongs to the field of novel functional molecules, and specifically relates to a reaction system, method and application of visible light-induced BODIPY photocage molecules to release aldehyde compounds. Background Art

[0002] Light-induced spatiotemporal controllable molecular release of photocage compounds (photocage molecules) is an important in situ release method for compounds. In general, photocage compounds are very stable in the system, and when they are placed under light conditions, the molecules change from the ground state to the excited state, and then a series of transformations occur to finally obtain the target molecule, thereby achieving molecular release. Because the reaction conditions are mild and efficient, and the release of compounds can be precisely controlled, this strategy is widely used in the biomedical field.

[0003] Among them, the boron pyrrole dimethine (BODIPY) molecule is widely used in the construction of photocage compounds due to its stable skeleton structure and excellent photochemical properties. At present, photocage molecules with BODIPY skeleton structure have been used to release target active compounds in vivo under light control, such as drug molecules, fluorescent molecules, peptides, nucleic acids, etc. However, the release method of the target compound in this system is currently limited to the method based on the cleavage of CO single bonds, and there is a lack of photocage molecular systems for aldehyde compounds.

[0004] In addition, although the photocage molecules based on the BODIPY skeleton structure have excellent optical properties, for the BODIPY photocage molecules that have been developed at this stage, there is no obvious difference in the skeleton structure before and after the release of the target molecules. Therefore, it is difficult to directly achieve in situ real-time monitoring of the release of target compounds through optical properties, which has caused certain limitations on its application.

[0005] Therefore, the development of BODIPY photocage molecules that can monitor the light-controlled release of aldehyde compounds in real time is of great practical significance for interdisciplinary subjects such as biology and materials chemistry. Summary of the invention

[0006] In order to overcome the defects of the above-mentioned prior art, an object of the present invention is to propose a reaction system for visible light-induced BODIPY photocage molecules to release aldehyde compounds, under which the olefin double bonds in the BODIPY photocage molecules are oxidatively broken to release aldehyde compounds and BODIPY-aldehyde compounds.

[0007] The second objective of the present invention is to propose a method for inducing the release of aldehyde compounds from the above-mentioned BODIPY photocage molecules by visible light. In an oxygen-containing environment, the BODIPY photocage molecule solution is irradiated with visible light, and the olefin double bond of the BODIPY photocage molecule is oxidatively broken to achieve rapid release of aldehyde compounds. At the same time, the generated BODIPY-aldehyde compound can be used as a biothiol fluorescent probe in a biological environment to monitor the release process in situ and in real time.

[0008] The third object of the present invention is to provide an application of the reaction system in which the BODIPY photocage molecule releases aldehyde compounds induced by visible light in drug monitoring, wherein the BODIPY-aldehyde compound is used as a biothiol fluorescent probe to monitor the release progress of the aldehyde compound.

[0009] The fourth object of the present invention is to provide an application of the reaction system in which the above-mentioned visible light-induced BODIPY photocage molecule releases aldehyde compounds in drug monitoring under biological environment, wherein the BODIPY photocage molecule is modified with a targeting group to construct a targeted prodrug molecule, and is used for the targeted and controllable release of aldehyde compounds in cells and organelles, and the aldehyde compounds are monitored in real time by using the BODIPY-aldehyde compound as a biothiol fluorescent probe.

[0010] This is achieved specifically through the following technical solutions:

[0011] A reaction system for releasing aldehyde compounds by BODIPY photocage molecules induced by visible light, characterized by comprising:

[0012] Reactants: BODIPY photocage molecules containing diene groups;

[0013] Reaction medium;

[0014] Reaction conditions: aerobic environment and light source irradiation conditions;

[0015] The reactants formed by the reaction system include: aldehyde compounds and BODIPY-aldehyde compounds.

[0016] Furthermore, the oxygen-containing environment is air condition or oxygen condition.

[0017] Specifically, the molecular structure of the BODIPY photocage is as follows:

[0018]

[0019] Among them, R is used to adjust the maximum absorption wavelength and quantum yield of the molecule. 1It is one or more of C1-C40 alkyl, C3-C12 cycloalkyl or C3-C12 cycloalkyl with a substituent, phenyl and substituted phenyl, benzyl and substituted benzyl, five-membered or six-membered heterocyclic aromatic group containing one or more oxygen, sulfur or nitrogen atoms, ester group, and monosaccharide, polysaccharide, polypeptide, and nucleic acid aptamer with a targeting functional structure;

[0020] R 2 , R 3 is one or more of hydrogen, C1-C40 alkyl, C3-C12 cycloalkyl or C3-C12 cycloalkyl with substituents, phenyl and substituted phenyl, benzyl and substituted benzyl, five-membered or six-membered heterocyclic aromatic group containing one or more oxygen, sulfur or nitrogen atoms, and ester group; the substituent on the C3-C12 cycloalkyl, the substituent on the phenyl, and the substituent on the benzyl are one or more of C1-C40 alkyl, C1-C40 alkoxy, halogen, nitro, ester group, or cyano group, and the number of substituents is 1-5; one or more of C1-C10 alkanoyl, C1-C10 alkoxycarbonyl, C1-C10 alkylsulfonyl, C1-C10 alkylphosphoryl, benzoyl and substituted benzoyl, phenoxycarbonyl and substituted phenoxycarbonyl, benzylsulfonyl and substituted benzylsulfonyl, or phenylphosphoryl and substituted phenylphosphoryl;

[0021] R 4 , R 5 It is one or more of C1-C40 alkyl, C3-C12 cycloalkyl or C3-C12 cycloalkyl with substituent, phenyl and substituted phenyl, benzyl and substituted benzyl, five-membered or six-membered heterocyclic aromatic group containing one or more oxygen, sulfur or nitrogen atoms, and ester group.

[0022] Furthermore, the reaction medium is at least one of water, methanol, ethanol, toluene, benzene, xylene, dichloromethane, dichloroethane, ether, tetrahydrofuran, dimethyl sulfoxide or N,N-dimethylformamide.

[0023] Furthermore, the light source is a full-spectrum light source or a laser light source with a maximum irradiance at 630 nm.

[0024] A method for releasing aldehyde compounds from BODIPY photocage molecules induced by visible light based on the above reaction system, characterized in that the specific steps include:

[0025] The BODIPY photocage molecule is dissolved in a reaction medium, and the BODIPY photocage molecule is irradiated by a light source in an oxygen-containing environment. After being irradiated by the light source, the BODIPY photocage molecule undergoes oxidative cleavage to release aldehyde compounds and BODIPY-aldehyde compounds.

[0026] Furthermore, the concentration of the photocage molecules is 0.1 μM-1M.

[0027] Furthermore, the light source irradiates for a duration greater than 1 minute.

[0028] Furthermore, the reaction temperature is room temperature, and the reaction pressure is normal pressure.

[0029] A method for monitoring the release of aldehyde compounds according to the above reaction system, wherein the BODIPY-aldehyde compound is used as a biothiol fluorescent probe to monitor the release progress of the aldehyde compound.

[0030] A method for drug monitoring in a biological environment based on the above reaction system, wherein the BODIPY photocage molecule is modified with a targeting group to construct a prodrug molecule with targeting, and is used for the targeted and controllable release of aldehyde compounds in cells and organelles, and the aldehyde compounds are monitored in real time by using the BODIPY-aldehyde compound as a biothiol fluorescent probe.

[0031] The present invention has the following advantages:

[0032] 1. The photocage molecules have good stability, high reaction conversion rate, mild reaction conditions, and a wide range of universal solvents;

[0033] 2. Photocage molecules are easy to synthesize;

[0034] 3. The photocage molecule can be used in the release reaction of molecules in biological systems, and the generated BODIY-aldehyde compound can be used as a fluorescent probe to detect the release progress;

[0035] 4. Compared with traditional aldehyde precursors, this photocage molecule is more stable in complex environments and can visualize the release process of aldehyde compounds. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings required for use in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0037] Figure 1 The synthetic route of the BODIPY photocage molecule of the embodiment of the present invention;

[0038] Figure 2 This is the hydrogen nuclear magnetic resonance spectrum of the photocage molecule BOD-dOH prepared in Example 1 of the present invention;

[0039] Figure 3 This is the carbon NMR spectrum of the photocage molecule BOD-dOH prepared in Example 1 of the present invention;

[0040] Figure 4 This is a high-resolution mass spectrum of the photocage molecule BOD-dOH prepared in Example 1 of the present invention;

[0041] Figure 5 This is the hydrogen nuclear magnetic resonance spectrum of the photocage molecule BOD-M-dOH prepared in Example 3 of the present invention;

[0042] Figure 6 This is the carbon NMR spectrum of the photocage molecule BOD-M-dOH prepared in Example 3 of the present invention;

[0043] Figure 7 This is a high-resolution mass spectrum of the photocage molecule BOD-M-dOH prepared in Example 3 of the present invention;

[0044] Figure 8 This is the hydrogen nuclear magnetic resonance spectrum of the photocage molecule BOD-Cl-dOH prepared in Example 4 of the present invention;

[0045] Fig. 9 This is the carbon nuclear magnetic resonance spectrum of the photocage molecule BOD-Cl-dOH prepared in Example 4 of the present invention;

[0046] Fig.10 This is a high-resolution mass spectrum of the photocage molecule BOD-Cl-dOH prepared in Example 4 of the present invention;

[0047] Fig.11 This is a hydrogen nuclear magnetic resonance spectrum of the photocage molecule BOD-dOH photolysis to release 4-hydroxybenzaldehyde in Example 5 of the present invention;

[0048] Fig.12 This is the nuclear magnetic resonance hydrogen spectrum of the fluorescent probe BOD-OH obtained by photolysis of the light cage molecule BOD-dOH in Example 5 of the present invention;

[0049] Fig.13 This is the carbon NMR spectrum of the fluorescent probe BOD-OH obtained by photolysis of the light cage molecule BOD-dOH in Example 5 of the present invention;

[0050] Fig.14 This is a high-resolution mass spectrum of the fluorescent probe BOD-OH obtained by photolysis of the light cage molecule BOD-dOH in Example 5 of the present invention;

[0051] Fig.15 This is the UV-visible absorption spectrum of the photocage molecule BOD-dOH prepared in Example 1 of the present invention;

[0052] Fig.16 This is the UV-visible absorption spectrum of the photocage molecule BOD-M-dOH prepared in Example 3 of the present invention;

[0053] Fig.17 The UV-visible absorption spectrum of the photocage molecule BOD-Cl-dOH prepared in Example 4 of the present invention;

[0054] Fig.18 This is the UV-visible absorption spectrum of the fluorescent probe BOD-OH obtained by photolysis of the light cage molecule BOD-dOH in Example 5 of the present invention;

[0055] Fig.19 This is a fluorescence spectrum of the light cage molecule BOD-dOH prepared in Example 1 of the present invention;

[0056] Fig. 20 This is a fluorescence spectrum of the light cage molecule BOD-M-dOH prepared in Example 3 of the present invention;

[0057] Fig.21 This is a fluorescence spectrum of the light cage molecule BOD-Cl-dOH prepared in Example 4 of the present invention;

[0058] Fig. 22 This is a fluorescence spectrum of the fluorescent probe BOD-OH obtained by photolysis of the light cage molecule BOD-dOH in Example 5 of the present invention;

[0059] Fig.23 This is a UV-visible absorption spectrum diagram of real-time monitoring of photolysis of the photocage molecule BOD-dOH in Example 15 of the present invention;

[0060] Fig.24 This is a fluorescence spectrum diagram of real-time monitoring of photolysis of the photocage molecule BOD-dOH in Example 15 of the present invention. DETAILED DESCRIPTION

[0061] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0062] The invention provides a BODIPY photocage molecule, which generates active oxygen under visible light conditions to oxidatively break its own olefin structure to obtain a target aldehyde compound and a BODIPY aldehyde skeleton structure, and the latter can be used as a biological thiol fluorescent probe in vivo.

[0063] The present invention can release various aldehyde compounds efficiently and accurately by light control. Compared with other aldehyde precursor compounds, the BODIPY photocage molecule has the advantages of stable structure, easy modification, sensitive response, and real-time monitoring. The present invention can also construct a targeted prodrug molecule by modifying the BODIPY photocage molecule with a targeting group, and use it for the targeted controlled release of aldehyde drugs in cells and organelles, and monitor it in real time through a biothiol fluorescent probe.

[0064] Experimental instruments and materials:

[0065] The nuclear magnetic resonance was measured by a Bruker400 nuclear magnetic resonance instrument, the liquid chromatography was an Agilent high-performance liquid chromatography, the high-resolution mass spectrometry was measured by an Agilent Thermo Q Exactive Focus, the UV-visible spectrum was measured by a UV-1800 UV-visible spectrophotometer, the fluorescence spectrum was measured by a FLS920 fluorescence spectrometer, the cell imaging was measured by a confocal fluorescence microscope ZEISS LSM 900, and the flow cytometric analysis was measured by a flow cytometer CytoFLEX / DxFLEX.

[0066] All materials used were obtained from commercial sources unless otherwise specified and were used directly without further purification.

[0067] Example 1 Synthesis of 4-hydroxybenzaldehyde photocage molecule BOD-dOH

[0068] Add BODIPY 1 (177 mg, 0.5 mmol), 4-hydroxybenzaldehyde (488 mg, 2.0 mmol), acetic acid (7 mL), piperidine (7 mL) to the reaction tube, and add 20 mL of anhydrous toluene under nitrogen protection. Then move the reaction tube to a 130°C oil bath heating stirrer and react overnight. After the reaction is completed, reduce pressure and concentrate until there is almost no solvent, separate by silica gel column chromatography, reduce pressure and concentrate, and vacuum dry to obtain a black solid BOD-dOH, 179 mg, with a yield of 64%.

[0069] The reaction equation is as follows:

[0070]

[0071] The nuclear magnetic resonance hydrogen spectrum, carbon spectrum and high-resolution mass spectrum of BOD-dOH are as follows Figure 2 , Figure 3 , Figure 4 As shown, where:

[0072] 1H NMR(400MHz,DMSO-d6)δ8.26(brs,1H),7.49–7.45(m,6H),7.38–7.30(m,4 H),7.24(d,J=8.6Hz,2H),6.90–6.86(m,6H),5.22(s,2H),4.76(s,2H),4. 68–4.55(m,4H),4.47(s,1H),3.98(d,J=4.7Hz,1H),3.83(d,J=4.5Hz,1H) ,3.64(d,J=10.6Hz,1H),3.57(s,1H),3.18(d,J=5.4Hz,2H),1.47(s,6H). 13 C NMR (101MHz, DMSO-d6) δ160.3,159.5,152.6,141.7,138.3,137.3,133.2,130.2,129.5,127.8,126.7,118.3,116.6,115.5,115.0,55.7,14.9.

[0073] HRMS m / z:[MH] – Calculate for C 34 H 28 O3N2BF2 – 561.21665,found:561.21665.

[0074] Example 2 Synthesis of intermediate BODIPY 3

[0075] Add BODIPY 2 (189 mg, 0.5 mmol), 4-hydroxybenzaldehyde (488 mg, 2.0 mmol), acetic acid (7 mL), piperidine (7 mL) to the reaction tube, and add 20 mL of anhydrous toluene under nitrogen protection. Then move the reaction tube to a 130°C oil bath heating stirrer and react overnight. After the reaction is completed, reduce pressure and concentrate until there is almost no solvent, separate by silica gel column chromatography, reduce pressure and concentrate, and vacuum dry to obtain black solid BODIPY, 100 mg, with a yield of 34%.

[0076] The reaction equation is as follows:

[0077]

[0078] Example 3 Synthesis of Mannose Receptor Targeted 4-Hydroxybenzaldehyde BODIPY Photocage Molecule

[0079] Add CuI (1.9 mg, 0.01 mmol), i-Pr2NEt (25.8 mg, 0.2 mmol), BODIPY 3 (64.5 mg, 0.11 mmol) and azide-substituted mannose Mannose-N3 (24.9 mg, 0.1 mmol) to the reaction tube, and add 1 mL of anhydrous acetonitrile under nitrogen protection. Then move the reaction tube to a 60°C oil bath heating stirrer and react overnight. After the reaction is completed, concentrate under reduced pressure until there is almost no solvent, separate by silica gel column chromatography, concentrate under reduced pressure, and dry in vacuo to obtain a black solid BOD-M-dOH, 18.2 mg, with a yield of 22%.

[0080] The reaction equation is as follows:

[0081]

[0082] The nuclear magnetic resonance hydrogen spectrum, carbon spectrum and high-resolution mass spectrum of BOD-M-dOH are as follows Figure 5 , Figure 6 , Figure 7 As shown, where:

[0083] 1 H NMR (400MHz, DMSO-d6) δ8.26(s,1H),7.49–7.46(m,6H),7.36–7.33(m,4H),7.24(d,J=8.6Hz,2H),6.92–6.87(m,6H),5.22(s,2H),4.76(s,2H ),4.66–4.58(m,4H),4.47(s,1H),3.98–3.97(m,1H),3.84–3.82(m,1H ),3.64(d,J=10.6Hz,1H),3.57(s,1H),3.19–2.17(m,2H),1.47(s,6H). 13 C NMR(101MHz,DMSO-d6)δ159.5,159.1,152.6,142.7,141.8,138.2,137.4,133.2,130.2,129.5,127.8, 127.1,125.6,118.3,116.6,115.8,115.5,100.3,74.7,71.3,70.5,67.2,65.4,61.7,61.6,49.9,14.9.

[0084] HRMS m / z:[M+Na] + Calculate for C 44 H 44 O9N5BF2Na + 858.30924, found:858.30988.

[0085] Example 4 Synthesis of endoplasmic reticulum-targeted 4-hydroxybenzaldehyde BODIPY photocage molecule

[0086] Add CuI (1.9 mg, 0.01 mmol), i-Pr2NEt (25.8 mg, 0.2 mmol), BODIPY 3 (64.5 mg, 0.11 mmol) and 1-chloro-3-azidopropane (11.9 mg, 0.1 mmol) to the reaction tube, and add 1 mL of anhydrous acetonitrile under nitrogen protection. Then move the reaction tube to a 60°C oil bath heating stirrer and react overnight. After the reaction is completed, concentrate under reduced pressure until there is almost no solvent, separate by silica gel column chromatography, concentrate under reduced pressure, and dry in vacuo to obtain a black solid BOD-Cl-dOH, 60.1 mg, with a yield of 86%.

[0087] The reaction equation is as follows:

[0088]

[0089] The nuclear magnetic resonance hydrogen spectrum, carbon spectrum and high-resolution mass spectrum of BOD-Cl-dOH are as follows Figure 8 , Fig. 9 , Fig.10 ,in:

[0090] 1 H NMR(400MHz,DMSO-d6)δ9.99(s,2H),8.33(s,1H),7.49-7.47(m,6H),7.35-7.33(m,4H),7.23(d,J=8.1Hz,2H) ,7.01–6.77(m,6H),5.23(s,2H),4.54(t,J=6.7Hz,2H),3.64(t,J=6.1Hz,2H),2.37–2.24(m,2H),1.46(s,6H).

[0091] Example 5 Release of 4-hydroxybenzaldehyde and biothiol fluorescent probe BOD-OH from light caged BOD-dOH under oxygen conditions in vitro

[0092] BOD-dOH (0.2 mmol), dichloromethane and methanol as a mixed solvent (1:1, 150 mL) were added to a 250 mL round-bottom flask with a branch tube, and then a normal pressure oxygen ball was connected to the branch tube. The reaction bottle was placed 2 cm above the LED lamp (20 W, 630 nm), and the reaction was placed under the light. The system turned from blue to red. The reaction was completed in 48 hours by TLC detection. After the system solvent was evaporated, 4-hydroxybenzaldehyde and BOD-OH were separated by column chromatography.

[0093] The reaction equation is as follows:

[0094]

[0095] The H NMR spectrum of 4-hydroxybenzaldehyde is Fig.11 , the nuclear magnetic resonance hydrogen spectrum, carbon spectrum and high-resolution mass spectrum of BOD-OH are as follows Fig.12 , Fig.13 , Fig.14 The UV-visible absorption spectrum is shown in Fig.18 The fluorescence emission spectrum is shown in Fig. 22 As shown, where:

[0096] 4-Hydroxybenzaldehyde: 1 H NMR (400MHz, DMSO-d6) δ10.60(s,1H),9.80(s,1H),7.78–7.76(m,2H),6.95–6.92(m,2H).

[0097] BOD-OH: 1 H NMR (400MHz, DMSO-d6) δ10.39(s,1H),10.16(s,1H),7.97(d,J=16.1Hz,1H),7.61(d,J=8.7Hz,2H),7.43–7.38(m,3 H),7.33(s,1H),7.15(d,J=8.7Hz,2H),6.93(d,J=8.6Hz,2H),6.83(s,1H),3.34(s,3H),1.55(s,3H),1.43(s,3H). 13 C NMR(101MHz,DMSO-d6)δ183.4,162.5,161.5,160.6,148.7,146.3,141.4,140.1,137.8,135 .1,133.8,131.3,130.0,126.9,125.8,122.7,120.1,117.0,115.2,114.1,55.8,15.6,14.2.

[0098] HRMS m / z:[MH] - Calculate for C 27 H 22 O3N2BF2 - 471.16970,found:471.16925.

[0099] Example 6 Release of 4-hydroxybenzaldehyde and biothiol fluorescent probe BOD-OH from light caged BOD-dOH under oxygen conditions in vitro

[0100] The concentration of BOD-dOH in Example 5 was changed to 2.5 μM, the reaction bottle was 10 mL, the reaction solvent was 1 mL of dichloromethane and methanol (1:1), and the rest was the same as Example 5. The reaction completion time was 3.5 hours as detected by liquid chromatography.

[0101] Example 7 Release of 4-hydroxybenzaldehyde and biothiol fluorescent probe BOD-OH from light caged BOD-dOH under in vitro air conditions

[0102] The oxygen condition in Example 6 was changed to air condition, and the rest was the same as Example 6. The reaction completion time monitored by liquid chromatography was 5.5 hours.

[0103] Example 8 Release of 4-hydroxybenzaldehyde and biothiol fluorescent probe BOD-OH from light caged BOD-dOH under nitrogen conditions in vitro

[0104] The oxygen condition in Example 6 was changed to nitrogen condition, and the rest was the same as Example 6. No release of 4-hydroxybenzaldehyde by the photocage molecule was monitored.

[0105] Example 9: Dimethyl sulfoxide as a reaction solvent to release 4-hydroxybenzaldehyde and biothiol fluorescent probe BOD-OH from light cage BOD-dOH

[0106] The methanol and dichloromethane mixed solvent in Example 7 was replaced by dimethyl sulfoxide, and the rest was the same as Example 7. The reaction completion time monitored by liquid chromatography was 11 hours.

[0107] Example 10 DMF as a reaction solvent to release 4-hydroxybenzaldehyde and biothiol fluorescent probe BOD-OH from photocage BOD-dOH

[0108] The methanol and dichloromethane mixed solvent in Example 7 was replaced by DMF, and the rest was the same as Example 7. The reaction completion time monitored by liquid chromatography was 7.5 hours.

[0109] Example 11 Tetrahydrofuran as a reaction solvent to release 4-hydroxybenzaldehyde and biothiol fluorescent probe BOD-OH from photocage BOD-dOH

[0110] The methanol and dichloromethane mixed solvent in Example 7 was replaced by DMF, and the rest was the same as Example 7. The reaction completion time monitored by liquid chromatography was 7 hours.

[0111] Example 12 Dichloromethane as a reaction solvent Light cage BOD-dOH releases 4-hydroxybenzaldehyde and biothiol fluorescent probe BOD-OH

[0112] The methanol and dichloromethane mixed solvent in Example 7 was replaced by dichloromethane. The rest was the same as Example 7. The reaction completion time monitored by liquid chromatography was 7 hours.

[0113] Example 13: Methanol as the reaction solvent to release 4-hydroxybenzaldehyde and biothiol fluorescent probe BOD-OH from light cage BOD-dOH

[0114] The methanol and dichloromethane mixed solvent in Example 7 was replaced by methanol, and the rest was the same as Example 7. The reaction completion time monitored by liquid chromatography was 6 hours.

[0115] Example 14 Water and methanol as reaction solvents to release 4-hydroxybenzaldehyde and biothiol fluorescent probe BOD-OH from light cage BOD-dOH

[0116] The methanol and dichloromethane mixed solvent in Example 7 was replaced with methanol and water (1:1, 1 mL), and the rest was the same as Example 7. The reaction completion time monitored by liquid chromatography was 6 hours.

[0117] Example 15 Water and methanol are used as reaction solvents, and light cage BOD-dOH releases 4-hydroxybenzaldehyde and biothiol fluorescent probe BOD-OH under oxygen conditions

[0118] The system in Example 14 was enlarged to 100 mL, and the reaction was carried out under oxygen conditions. The rest was the same as in Example 14. The reaction was completed in 3 hours using liquid chromatography. The UV-visible absorption spectrum and fluorescence spectrum of the photolysis process are shown in FIG. Fig.23 , Fig.24 .

[0119] Example 16 Release of 4-hydroxybenzaldehyde and biothiol fluorescent probe BOD-OH from light caged BOD-dOH in HeLa cell environment

[0120] The light cage BOD-dOH (10 μM) was incubated with HeLa cells for 12 hours, and after changing the solution and washing three times, the cells were illuminated with LED light (630 nm, 24 W) for 5 minutes, and the intracellular release of the light cage molecules was analyzed by flow cytometry and confocal fluorescence microscopy. According to the flow cytometric analysis, 15% of the cells could detect the production of biothiol fluorescent probes.

[0121] Example 17 Endoplasmic reticulum-targeted photocage BOD-Cl-dOH releases 4-hydroxybenzaldehyde and biothiol fluorescent probes in HeLa cell environment

[0122] The light cage BOD-Cl-dOH (10μM) and endoplasmic reticulum dye (ER-Tracker Blue-White DPX) were co-incubated with HeLa cells for 2 hours. After changing the solution and washing three times, the cells were illuminated with LED light (630nm, 24W) for 5 minutes, and the intracellular release of the light cage molecules was analyzed by flow cytometry and confocal fluorescence microscopy imaging. According to flow cytometry analysis, 12% of the cells could detect the generation of biothiol fluorescent probes. Confocal fluorescence microscopy co-localization imaging showed that the light cage molecule had good endoplasmic reticulum targeting.

[0123] Example 18 Rapid enrichment of mannose receptor-targeted photocage molecules and release of 4-hydroxybenzaldehyde and biothiol fluorescent probes from BOD-M-dOH in the HepG2 cell environment

[0124] The light cage BOD-M-dOH (2μM) was co-incubated with HepG2 cells for 1 hour. After changing the solution and washing 3 times, the cells were illuminated with LED light (630nm, 24W) for 5 minutes, and the intracellular release of light cage molecules was analyzed by flow cytometry and confocal fluorescence microscopy. According to flow cytometry analysis, 80% of the cells were detected to have enriched BOD-M-dOH before illumination. After illumination, 10% of the cells were detected to have generated biothiol fluorescent probes.

[0125] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for releasing aldehyde compounds by using BODIPY photocage molecules induced by visible light, characterized in that: include: Reactants: BODIPY photocage molecules; Reaction medium; Reaction conditions: aerobic environment and light source irradiation conditions; The products formed by the method are: aldehyde compounds and BODIPY-aldehyde compounds; The molecular structure of the BODIPY photocage is shown below: The light source is a laser light source with a maximum irradiance of 630 nm.

2. The method according to claim 1, characterized in that The aerobic environment is air condition or oxygen condition.

3. The method according to claim 1, characterized in that The reaction medium is at least one of water, methanol, ethanol, toluene, benzene, xylene, dichloromethane, dichloroethane, ether, tetrahydrofuran, dimethyl sulfoxide or N,N-dimethylformamide.

4. The method according to claim 1, characterized in that: The specific steps include: The BODIPY photocage molecules are dissolved in a reaction medium, and the BODIPY photocage molecules are irradiated by a light source in an oxygen-containing environment. After being irradiated by the light source, the BODIPY photocage molecules undergo oxidative cleavage to release aldehyde compounds and BODIPY-aldehyde compounds.

5. The method according to claim 4, characterized in that The concentration of the photocage molecules is 0.1 μM-1M.

6. The method according to claim 4, characterized in that The light source irradiates for longer than 1 minute.

7. The method according to claim 4, characterized in that The reaction temperature is room temperature, and the reaction pressure is normal pressure.

8. Use of the method according to claim 1 in preparing a reagent for monitoring the release of aldehyde compounds, wherein the BODIPY-aldehyde compound is used as a biothiol fluorescent probe to monitor the release progress of the aldehyde compound.

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