CN110357817A - A kind of invertibity detects pyroracemic aldehyde and glyoxal fluorescence probe and its preparation method and application - Google Patents

Abstract

The invention discloses a kind of fluorescent probes for reversible detection of glyoxal and glyoxal and a preparation method and application thereof. The method comprises: refluxing 4-bromo-1,8-naphthalene anhydride and amine in ethanol for 3.5 hours to obtain N ‑R1‑4‑bromo‑1,8‑naphthalimide, then, under nitrogen protection, N‑R1‑4‑bromo‑1,8‑naphthalimide and guanidine in ethylene glycol methyl ether at 100 Stir overnight at ℃ to obtain a fluorescent probe compound for reversible detection of glyoxal and glyoxal. Using 1,8-naphthylimide as a fluorophore, a guanidine group is used to react with glyoxal and glyoxal to form a reversible dialdehyde. Hydroxyimidazolidine, the excited state intramolecular charge transfer effect is increased, resulting in strong fluorescence. Therefore, the fluorescent probe of the present invention can rapidly and reversibly detect glyoxal and glyoxal.

Description

A kind of fluorescent probe for reversible detection of glyoxal and glyoxal and its preparation method and application

technical field

The invention relates to the technical field of organic synthesis and analytical chemistry, in particular to a type of fluorescent probes for reversible detection of glyoxal and glyoxal and a preparation method and application thereof.

Background technique

Reactive carbonyl compounds, such as formaldehyde, acetaldehyde, glyoxal, glyoxal, acrolein, etc., play an important role in living systems. Due to the high reactivity of these compounds with DNA and proteins, these active carbonyl compounds are related to the occurrence and development of other chronic diseases such as diabetes, cancer, obesity, Alzheimer's disease, and aging. Among them, glyoxal and glyoxal, as active carbonyl compounds, can induce advanced glycation end products.

Glycoaldehyde mainly reacts with lysine, arginine, and cysteine residues to produce glycation end products. This reaction mainly affects different proteins, such as insulin, hemoglobin and growth factors, and can also cause DNA damage. damage, resulting in cytotoxicity. It has been reported that the content of methylglyoxal in normal human serum is 0.4-1.0 μM, while that of diabetic patients can reach 2.2-3.8 μM (Analytical Letters. 2006, 39(10): 2205-2215.). To be able to better understand the function of these reactive carbonyl compounds under physiological and pathological conditions, methods to selectively detect and monitor the levels of reactive carbonyl compounds in biological samples need to be developed. Most of the detection of glyoxal is the reaction of o-phenylenediamine and glyoxal to generate stable groups. For example, HPLC, LC-MS, and GC-MS are all used to detect glyoxal based on this principle. However, these detection methods all require cell lysis and are not suitable for application in complex living systems. Fluorescence detection has received extensive attention due to its simplicity, sensitivity, and biocompatibility, which can detect analytes in living systems.

So far, the fluorescence detection of glyoxal all use o-phenylenediamine as the detection group, which generates fluorescence after irreversible reaction with glyoxal. This method has shortcomings in selectivity, reaction speed and dynamic detection. Therefore, it is necessary and challenging to develop a new group for the detection of glyoxal.

SUMMARY OF THE INVENTION

In the present invention, 1,8-naphthalimide is used as a fluorophore, and the guanidine group is reacted with glyoxal and glyoxal to form a reversible dihydroxyimidazolidine, and the ICT effect is increased, thereby generating strong fluorescence. Therefore, the fluorescent probe of the present invention can rapidly and reversibly detect glyoxal and glyoxal.

In order to realize the main purpose of the present invention, the present invention provides:

A fluorescent probe for reversible detection of glyoxal and glyoxal, the fluorescent probe has the following general formula I or general formula II or general formula III:

Wherein, R1 and R2 are both alkyl groups; n is a natural number of 1, 2 or 3.

Preferably, the R1 and R2 are each independently methyl, ethyl and propyl.

Preferably, the formula I is the following formula I-1:

Preferably, the formula II is the following formula II-1:

Preferably, the formula III is the following formula III-1:

The method of the fluorescent probe shown in the above general formula I is synthesized by the following route:

The method comprises the steps of refluxing 4-bromo-1,8-naphthalene anhydride and amine in ethanol for 3.5 h to obtain N-R1-4-bromo-1,8-naphthalimide, and then, under nitrogen protection, N-R1-4-bromo-1,8-naphthalimide -R1-4-bromo-1,8-naphthalimide and guanidine were stirred overnight at 100°C in ethylene glycol methyl ether to obtain fluorescent probe compounds for reversible detection of glyoxal and glyoxal.

Wherein, the molar ratio of 4-bromo-1,8-naphthalene anhydride to amine is 1:5, and the molar ratio of intermediate NR ¹ -4-bromo-1,8-naphthalimide to guanidine is 1:5.

The method of the fluorescent probe shown in the above general formula II is synthesized by the following route:

The method comprises that under nitrogen protection, NR ² -4-bromo-1,8-naphthalimide and guanidine are stirred in ethylene glycol methyl ether at 100° C. overnight to obtain reversible detection of glyoxal and ethylene glycol. Aldehyde fluorescent probe compound, wherein, the molar ratio of NR ² -4-bromo-1,8-naphthalimide and formidine is 1:5.

The method of the fluorescent probe shown in the above general formula III is synthesized by the following route:

The method comprises that under nitrogen protection, compound 1 and guanidine are stirred in ethylene glycol methyl ether at 100° C. overnight to obtain a fluorescent probe compound for reversible detection of glyoxal and glyoxal, wherein the compound 1 and guanidine are combined with guanidine. The molar ratio is 1:5. Preferably, the present invention also discloses the application of the fluorescent probe prepared by the above method in detecting glyoxal and glyoxal in solution, cells or organisms.

The fluorescent probe for reversible detection of glyoxal and glyoxal according to the present invention, after the probe is combined with glyoxal or glyoxal, the ICT effect is greatly increased, resulting in an increase in fluorescence intensity. The identification response is as follows:

Fluorescent probes: have characteristic fluorescence in the ultraviolet-visible-near-infrared region, and their fluorescence properties (excitation and emission wavelengths, intensity, lifetime, polarization, etc.) can be changed with the properties of the environment, such as polarity, refractive index, viscosity, etc. A class of fluorescent molecules that are sensitively changed; small molecules that interact with nucleic acids (DNA or RNA), proteins or other macromolecular structures non-covalently to change one or more fluorescent properties. It can be used to study the properties and behavior of macromolecular substances.

Compared with the prior art, the present invention has beneficial technical effects:

(1) Reversible, fluorescent probes for reversible detection of glyoxal and glyoxal, realizing rapid and reversible detection of glyoxal and glyoxal levels in cells and serum;

(2) High sensitivity, the fluorescent probe of the invention can selectively react with glyoxal and glyoxal quickly and specifically to generate a product with strong fluorescence. Compared with other common aldehydes, amino acids and metal ions, the present invention The fluorescent probe showed high selectivity and sensitivity for glyoxal and glyoxal;

(3) The reaction time is fast, and the reaction of the fluorescent probe of the present invention with glyoxal and glyoxal can produce a product with strong fluorescence within 15 minutes at room temperature;

(4) It is widely used and has great potential. The probe has good optical stability and specific response to glyoxal and glyoxal. It can be used to detect the levels of glyoxal and glyoxal in the serum of diabetic model animals, which has potential practical Value;

(5) The preparation has obvious advantages, the preparation method of the fluorescent probe is simple, and the yield is high.

Description of drawings

Fig. 1 is the hydrogen nuclear magnetic resonance spectrum of probe I-1;

Fig. 2 is the hydrogen nuclear magnetic resonance spectrum of probe II-1;

Fig. 3 is the hydrogen nuclear magnetic resonance spectrum of probe III-1;

Fig. 4 is the mass spectrum of probe I-1;

Fig. 5 is the mass spectrum of probe II-1;

Fig. 6 is the mass spectrum of probe III-1;

Fig. 7 is the fluorescence change of probe I-1 with the action of different concentrations of acetaldehyde;

Fig. 8 is the fluorescence change of probe I-1 with the action of different concentrations of glyoxal;

Fig. 9 is the fluorescence change over time of the interaction of probe I-1 with acetaldehyde;

Figure 10 is the fluorescence change of probe I-1 interacting with glyoxal over time;

Figure 11 is the reversibility study of the reaction between probe I-1 and acetaldehyde;

Figure 12 is a reversibility study of the reaction between probe I-1 and glyoxal;

Figure 13 is a selectivity study of probe I-1;

Figure 14 is the fluorescence change of probe II-1 with the action of different concentrations of glyoxal;

Figure 15 is a selectivity study of probe II-1;

Figure 16 is a co-localized fluorescence image of probe I-1 with glyoxal and glyoxal.

Detailed ways

The present invention will be further described below through specific embodiments, and it should be understood that the following description is only for illustrating the present invention, and does not limit the content of the present invention.

The raw materials and equipment used in the examples are well known to those skilled in the art, and are commercially available or easily obtained or prepared.

Example 1

Synthesis of Fluorescent Probe I-1:

4-Bromo-1,8-naphthalene anhydride (1.1 g, 4 mmol) was suspended in 10 mL of ethanol, n-propylamine (1.18 g, 20 mmol) was added dropwise, and the reaction was heated under reflux for 3.5 h. Put it to room temperature, filter, wash with ethanol three times, and dry the filter cake to obtain the product, N-propyl-4-bromo-1,8-naphthalimide as a light yellow solid (1.02 g, 80.5%). Under the protection of nitrogen, compound N-propyl-4-bromo-1,8-naphthalimide (382 mg, 1.2 mmol) was suspended in 5 mL of ethylene glycol methyl ether, and then guanidine (355 mg, 6 mmol) was dissolved in In 3 mL of ethylene glycol methyl ether, a solution of guanidine in ethylene glycol methyl ether was slowly added dropwise to the suspension of N-propyl-4-bromo-1,8-naphthalimide at room temperature. The reaction mixture was stirred at 100°C overnight. Ethyl acetate and water were added for extraction. The organic layer was dried over anhydrous sodium sulfate, filtered, spin-dried, and subjected to column chromatography (dichloromethane:methanol=10:1, volume ratio) to obtain compound I-1 (138 mg, 39%) as an orange-red solid.

The structural characterization of compound I-1 by H NMR spectrum and high-resolution mass spectrometry is shown in FIG. 1 and FIG. 4 . Among them, the hydrogen nuclear magnetic resonance spectrum data: ¹ HNMR (400MHz, DMSO-d ₆ ) δ8.56 (dd, J=8.2, 1.1 Hz, 1H), 8.44 (dd, J=7.4, 1.1 Hz, 1H), 8.32 ( d, J＝8.2Hz, 1H), 7.72(dd, J＝8.2, 7.4Hz, 1H), 7.33(d, J＝8.2Hz, 1H), 6.70(bs, 4H), 4.00(m, 2H), 1.63(h, J=7.4Hz, 2H), 0.90(t, J=7.4Hz, 3H);

High resolution mass spectrometry (ESI-HRMS): m/z calculated according to formula C ₁₆ H ₁₇ N ₄ O ₂ [M+H] ⁺ peak calcd. 297.1346 found 297.1353.

Example 2

Synthesis of Fluorescent Probe II-1:

Under the protection of nitrogen, compound N-propyl-4-bromo-1,8-naphthalimide (100 mg, 0.314 mmol) was suspended in 1 mL of ethylene glycol methyl ether, and then formin (115 mg, 1.57 mmol) was suspended in 1 mL of ethylene glycol methyl ether. ) was dissolved in 1 mL of ethylene glycol methyl ether, and the ethylene glycol methyl ether solution of formin was slowly added dropwise to the suspension of N-propyl-4-bromo-1,8-naphthalimide at room temperature. . The reaction mixture was stirred at 100°C overnight. Ethyl acetate and water were added for extraction. The organic layer was dried over anhydrous sodium sulfate, filtered, spin-dried, and subjected to column chromatography (dichloromethane:methanol=10:1, volume ratio) to obtain compound II-1 (27 mg, 28%) as an orange-red solid.

The structural characterization of compound II-1 by H NMR and high-resolution mass spectrometry is shown in Figure 2 and Figure 5: ¹ HNMR (400MHz, DMSO-d ₆ )δ8.63 (dd, J=8.2, 1.2 Hz, 1H) ,8.40(dd,J＝7.4,1.2Hz,1H),8.24(d,J＝8.3Hz,1H),7.62(dd,J＝8.2,7.4Hz,1H),7.08(d,J＝8.3Hz, 1H), 6.41(bs, 1H), 6.07(bs, 2H), 3.99(m, 2H), 2.82(s, 3H), 1.62(h, J=7.4Hz, 2H), 0.90(t, J=7.4 Hz,3H);

High resolution mass spectrometry (ESI-HRMS): m/z calculated according to formula C ₁₇ H ₁₉ N ₄ O ₂ [M+H] ⁺ peak 311.1503 found 311.1510.

Example 3

Synthesis of Fluorescent Probe III-1:

Under the protection of nitrogen, compound a (947 mg, 2 mmol) was suspended in 15 mL of ethylene glycol methyl ether, and then guanidine (591 mg, 10 mmol) was dissolved in 3 mL of ethylene glycol methyl ether, and the guanidine was slowly dissolved at room temperature. Ethylene glycol methyl ether solution was added dropwise to the suspension of a. The reaction mixture was stirred at 100°C overnight. Ethyl acetate and water were added for extraction. The organic layer was dried over anhydrous sodium sulfate, filtered, spin-dried, and subjected to column chromatography (dichloromethane:methanol:triethylamine=200:20:1) to obtain compound III-1 (160 mg, 18%), which was orange-red solid.

The structural characterization of compound III-1 is shown in Figure 3 and Figure 6: ¹ HNMR (400MHz, DMSO-d ₆ )δ8.61 (dd, J=8.3, 1.2 Hz, 1H), 8.36 (dd, J=7.3, 1.2 Hz,1H),8.22(d,J=8.3Hz,1H),7.72(bs,1H),7.64–7.59(m,3H),7.26(d,J=8.1Hz,2H),7.12(d,J =8.3Hz,1H),6.17(bs,4H),4.09(t,J=6.7Hz,2H),3.03(bs,2H),2.28(s,3H);

High resolution mass spectrometry (ESI-HRMS): m/z calculated according to _{formula C22H22N5O4S} [M ₊ H] ⁺ peak calcd. _452.1387 found 452.1393.

Example 4

Fluorescence Spectrum Changes of Fluorescent Probe I-1 Reacting with Different Concentrations of Glyoxal and Glyoxal

Dissolve the fluorescent probe I-1 prepared in Example 1 in dimethyl sulfoxide (DMSO) to prepare a fluorescent probe mother solution with a concentration of 1 mM; add a 40% mass fraction of glyoxal solution to distilled water , to prepare glyoxal mother solutions with concentrations of 10 mM and 100 mM; 8.8 M glyoxal solution was added to distilled water to form glyoxal mother solutions with concentrations of 10 M and 100 M. Calculate the required PBS aqueous solution (10mM, pH=7.4) according to the concentration of the fluorescent probe and glyoxal/glyoxal and add it to a 1cm×1cm quartz cuvette (volume 3.5mL), take 6μL of the fluorescent probe stock solution and add it To the PBS aqueous solution, add glyoxal/glyoxal stock solutions (0–500 μM) of different concentrations to prepare a total of 3 mL of the test solution with a probe concentration of 2 μM. After 30 minutes of reaction, the fluorescence spectrum changes of the fluorescent probe I-1 reacted with different concentrations of glyoxal/glyoxal were measured by a fluorescence spectrometer (excitation wavelength was 425 nm). Fluorescence spectral changes are shown in Figures 7 and 8. It can be seen that the fluorescence peak of the probe solution at 564 nm is gradually enhanced with the increasing concentration of glyoxal/glyoxal.

Example 5

Changes of fluorescence spectra of fluorescent probe I-1 reacting with glyoxal and glyoxal with time

Calculate the required PBS aqueous solution (10mM, pH=7.4) according to the concentration of the fluorescent probe and glyoxal/glyoxal and add it to a 1cm×1cm quartz cuvette (volume 3.5mL), take 6μL of Example 1 The fluorescent probe stock solution was added to the PBS aqueous solution, and then 60 μL (10 mM) glyoxal/glyoxal stock solution was added to prepare a total of 3 mL of test solution with a probe concentration of 2 μM and a glyoxal/glyoxal concentration of 200 μM. Using 425nm as the excitation wavelength, the fluorescence spectrum changes with time were tested. As shown in Figures 9 and 10, the fluorescence peak at 564 nm gradually increased with time.

Example 6

Study on the reversibility of fluorescent probe Ⅰ-1 with glyoxal and glyoxal.

Calculate the required PBS aqueous solution (10mM, pH=7.4) according to the concentration of fluorescent probes and glyoxal/glyoxal and add it to a 1cm×1cm quartz cuvette (volume 3.5mL), take aminoguanidine (AG) 110mg To aqueous solutions, 10 mM and 100 mM stock solutions were prepared. Take 6 μL of the fluorescent probe stock solution in Example 1 and add it to the PBS aqueous solution, then add 60 μL (10 mM) of glyoxal/pyruvaldehyde stock solution to prepare a probe concentration of 2 μM, and then add different concentrations of aminoguanidine hydrochloride (150 μM- 1 mM) of the test solution in a total of 3 mL. Among them, aminoguanidine is used as inhibitor. Using 425nm as the excitation wavelength, the fluorescence spectrum changes with different inhibitor concentrations were tested, as shown in Figures 11 and 12, it can be found that after adding the inhibitor, no matter whether the mother solution is glyoxal or glyoxal, the fluorescence intensity at 564nm does decrease, and The magnitude of the decrease increases with increasing inhibitor concentration.

Example 7

Selectivity of Probe I-1 to Different Interfering Analytes

Calculate the required PBS aqueous solution (10mM, pH=7.4) according to the concentrations of fluorescent probes and different interfering analytes, add them to a 1cm×1cm quartz cuvette (volume 3.5mL), and take 6μL of the fluorescent probe in Example 2. The needle stock solution was added to the aqueous PBS solution, followed by the addition of 200 μM of different analytes: formaldehyde, salicylaldehyde, benzaldehyde, triphosgene, nitric oxide donor NOC-18, glyoxal, glyoxal. A total of 3 mL of test solutions with a probe concentration of 2 μM and a concentration of 200 μM of different interfering analytes were prepared, while leaving a blank test sample with only probes added. After 30 minutes of reaction, the fluorescence emission intensity at 564 nm (excitation wavelength of 425 nm) of different samples was measured with a fluorescence spectrometer. As shown in Figure 13, relative to the blank test solution, the fluorescence of the test solutions of glyoxal and glyoxal increased significantly, while the fluorescence enhancement of other analytes was not much. The experimental results show that the fluorescent probe I-1 has good selectivity for glyoxal and glyoxal.

Example 8 Fluorescence spectrum changes of probe II-1 with different concentrations of acetaldehyde

Dissolve the fluorescent probe II-1 prepared in Example 2 in DMSO to prepare a fluorescent probe II stock solution with a concentration of 1 mM; calculate the required PBS aqueous solution (10 mM, pH =7.4) was added to a 1cm×1cm quartz cuvette (volume 3.5mL), 6μL of fluorescent probe stock solution was added to the PBS aqueous solution, and then different concentrations of glyoxal stock solution (0–1200μM) were added to configure the probe concentration A total of 3 mL of 2 μM test solution. After 30 minutes of reaction, the fluorescence spectrum changes of fluorescent probe II-1 reacted with different concentrations of glyoxal were tested by a fluorescence spectrometer (excitation wavelength was 391 nm). Fluorescence spectrum changes are shown in Figure 14. It can be seen that the fluorescence peak of the probe solution at 564 nm is gradually enhanced with the gradual increase of the concentration of glyoxal.

Example 8

Selectivity of Probe II-1 for Different Interfering Analytes

Calculate the required PBS aqueous solution (10mM, pH=7.4) according to the concentrations of fluorescent probes and different interfering analytes, add them to a 1cm×1cm quartz cuvette (volume 3.5mL), and take 6μL of the fluorescent probe in Example 2. The needle stock solution was added to the PBS aqueous solution, and then 400 μM of different analytes were added: formaldehyde, glyoxal, acetaldehyde, glyoxal, o-phthalaldehyde, glyoxylic acid, benzaldehyde, glutathione, glucose, cysteine Amino acid, hydrogen peroxide, potassium ion, calcium ion, sodium ion, copper ion, zinc ion, aluminum ion, prepared into a test solution with a probe concentration of 2 μM and a concentration of 400 μM of different interfering analytes in a total of 3 mL, while leaving a Blank test sample with probe added. After 60 minutes of reaction, the fluorescence emission intensity at 564 nm (excitation wavelength of 391 nm) of different samples was measured with a fluorescence spectrometer. As shown in Figure 15, relative to the blank test solution, the fluorescence of the test solutions of glyoxal and formaldehyde increased significantly, while the fluorescence of other analytes was not enhanced much. The experimental results show that the fluorescent probe II-1 has good selectivity for glyoxal and glyoxal.

Example 9

Colocalization Fluorescence Imaging of Probe I-1 with Pyruvaldehyde and Glyoxal in HeLa Cells

HeLa cells were seeded in T-25 cell culture flasks in DMEM medium (containing phenol red, Gibco/Invitrogen) containing 10% fetal bovine serum and 1% glutamine for 48 hours at 37°C. The day before the experiment, cells were transferred to 3 Nunc 35mm glass bottom cell culture dishes (ThermoScientific) to allow cells to adhere. Probe I-1 was diluted in PBS buffer and added to the cell culture medium at a final concentration of 5 μM. After 1 hour incubation in the cell incubator at 37°C, the medium containing probe I-1 was removed. After washing twice with PBS buffer, cell culture medium containing 20 μM glyoxal or glyoxal was added to the glyoxal and glyoxal dishes. The same amount of fresh medium was added to the control (probe dishes only). After 30 min incubation at 37°C in a cell incubator, cells were imaged on a Leica TCSSP8 equipped with a 40× objective and a photomultiplier tube gain of 800 (excitation and emission wavelengths λ _ex/em = 405/530-590 nm) . After the first imaging step, 5 mM acetylcysteine was added to the cell culture medium of the glyoxal and glyoxal dishes. Cells were then incubated for an additional 30 min at 37°C in a cell incubator. At the end of the incubation, cell images were captured again on a Leica TCSSP8 equipped with a 40× objective and a photomultiplier tube gain of 800 (excitation and emission wavelengths λ _ex/em = 405/530-590 nm). As shown in FIG. 16 , the first group had no fluorescence, the second and fourth groups had obvious fluorescence, and the third and fifth groups were less fluorescent than the second and fifth groups. The experimental results show that the fluorescent probe Ⅰ-1 can reversibly perform fluorescence imaging on glyoxal and glyoxal in HeLa cells.

The above embodiments are only preferred examples of the present invention, and are not intended to limit the scope of implementation of the present invention. Therefore, any equivalent changes or modifications made according to the structures, features and principles described in the scope of the patent application of the present invention shall be included in the present invention. within the scope of the invention patent application.

Claims (15)

1. a fluorescent probe for reversible detection of glyoxal and glyoxal, wherein the fluorescent probe has the following general formula I or general formula II or general formula III: Wherein, R1 and R2 are both alkyl groups; n is a natural number of 1, 2 or 3. 2 . The fluorescent probe for reversibly detecting glyoxal and glyoxal according to claim 1 , wherein the R1 and R2 are each independently methyl, ethyl or propyl. 3 . 3. reversible detection glyoxal and glyoxal fluorescent probe as claimed in claim 2 is characterized in that, described formula I is following formula I-1: 4. The reversible detection glyoxal and glyoxal fluorescent probe of claim 2, wherein the formula II is the following formula II-1: 5. reversible detection glyoxal and glyoxal fluorescent probe as claimed in claim 1, is characterized in that, described formula III is following formula III-1: 6. a method for preparing the fluorescent probe shown in general formula I in claim 1, is characterized in that, this method is synthesized by following route: 7. A method for preparing the fluorescent probe shown in general formula II in claim 1, wherein the method is synthesized by the following route: 8. A method for preparing the fluorescent probe shown in the general formula III in claim 1, wherein the method is synthesized by the following route: 9. The method for preparing a fluorescent probe for reversible detection of glyoxal and glyoxal as claimed in claim 6, wherein the method comprises refluxing 4-bromo-1,8-naphthalene anhydride and amine in ethanol for 3.5 After h, N-R1-4-bromo-1,8-naphthalimide was obtained. Then, under nitrogen protection, N-R1-4-bromo-1,8-naphthalimide and guanidine were dissolved in ethylene glycol. Methyl ether was stirred at 100°C overnight to obtain fluorescent probe compounds for reversible detection of glyoxal and glyoxal. 10. The method for preparing a fluorescent probe for reversible detection of glyoxal and glyoxal as claimed in claim 9, wherein the molar ratio of the 4-bromo-1,8-naphthalene anhydride to the amine is 1:5 , the molar ratio of intermediate N-R1-4-bromo-1,8-naphthalimide and guanidine is 1:5. 11. The method for preparing reversible detection of glyoxal and glyoxal fluorescent probes as claimed in claim 7, characterized in that, the method comprises, under nitrogen protection, N-R2-4-bromo-1,8 - Naphthalimide and formidine were stirred in ethylene glycol methyl ether at 100°C overnight to obtain fluorescent probe compounds for reversible detection of glyoxal and glyoxal. 12. The method for preparing a fluorescent probe for reversible detection of glyoxal and glyoxal as claimed in claim 11, wherein the moles of N-R2-4-bromo-1,8-naphthalimide and formidine The ratio is 1:5. 13. The method for preparing a fluorescent probe for reversible detection of glyoxal and glyoxal as claimed in claim 12, wherein the method comprises, under nitrogen protection, compound (1) and guanidine in ethylene glycol methyl ether Stir overnight at 100°C to obtain fluorescent probe compounds for reversible detection of glyoxal and glyoxal. 14 . The method for preparing a fluorescent probe for reversible detection of glyoxal and glyoxal according to claim 13 , wherein the molar ratio of the compound (1) to guanidine is 1:5. 15 . 15. A use of the fluorescent probe according to any one of claims 1-14 in detecting glyoxal and glyoxal in a solution, a cell or a living organism.