A fluorescent probe based on 2,5-dioctyl-3,6-diphenylpyrrolopyrrole dione and its preparation method

Abstract

The present invention belongs to the field of probe detection technology, and in particular to a fluorescent probe based on 2,5-dioctyl-3,6-diphenylpyrrolopyrrole diketo and a preparation method thereof. In order to overcome the technical defects of existing instruments that are expensive and complicated to operate, the present application provides a new fluorescent probe based on 2,5-dioctyl-3,6-diphenylpyrrolopyrrole diketo. Aldehyde groups are introduced into 2,5-dioctyl-3,6-diphenylpyrrolopyrrole diketo, and different fluorescent probes are constructed by reacting the aldehyde groups with different recognition groups to achieve the detection of specific substances.

Description

Technical Field

[0001] The present application belongs to the field of fluorescent probes, and specifically relates to a fluorescent probe based on 2,5-dioctyl-3,6-diphenylpyrrolopyrroledione and a preparation method thereof. Background Art

[0002] The advantage of fluorescent probes in the detection field lies in their ability to identify specific chemicals with extremely high sensitivity and selectivity, and to convert this microscopic identification information into intuitive fluorescent signals, allowing for the accurate detection of target substances in complex biological samples. Fluorescent probes are easy to use, cost-effective, and suitable for widespread application in laboratories. In addition, fluorescence imaging technology has a place in the field of biomedical imaging due to its high spatial resolution and ability to provide precise positioning information. The multi-channel detection capability of fluorescent probes allows for the simultaneous detection of multiple targets in the same reaction system, improving the efficiency and accuracy of experiments.

[0003] Diketopyrrolopyrrole (DPP) is a conjugated organic compound composed of two pyrrole rings connected by a diketone group. It exhibits excellent photoelectric properties, high molar absorptivity, good charge transport capabilities, chemical structure modifiability, and good photostability and thermal stability. Due to its unique chemical and physical properties, it exhibits broad application prospects in scientific research, including applications as an organic luminescent material in bioimaging and chemical sensors, potential as a photosensitizer and photothermal agent in tumor therapy, use as a high-performance material in organic field-effect transistors and organic photovoltaic devices, and potential applications in sensors, light-absorbing materials, lithium-sulfur batteries, and optoelectronic devices.

[0004] Mercury ions (Hg 2+ ) is one of the most toxic heavy metal pollutants, which can cause serious environmental and health problems. 2+ It is easily absorbed by plants and accumulated through the food chain and converted into more dangerous organic mercury, especially methylmercury. This form of mercury is extremely destructive to the human nervous system and may cause symptoms such as neurasthenia, memory loss, poor sleep quality, and emotional instability. Mercury also has direct damage to the kidneys. High concentrations of mercury can directly damage renal tubular cells and cause acute tubular necrosis. Long-term exposure may also lead to nephrotic syndrome.

[0005] Endogenous hypochlorous acid (HClO) in organisms mainly comes from the oxidation reaction of hydrogen peroxide and chloride ions catalyzed by myeloperoxidase (MPO) in neutrophils. Neutrophils, as an important part of the immune system, participate in the body's immune defense mechanism. HClO and hypochlorite ions (ClO -) is essential for clearing invading bacteria and viruses, participating in cell differentiation and migration and other life activities when maintaining normal concentration. However, when endogenous ClO - When the content is too high, it may damage nucleic acids, proteins, lipids, etc. in cells through oxidation or chlorination, causing cell apoptosis and tissue damage, which is related to the occurrence of diseases such as acute lung injury, cardiovascular disease, neurodegenerative diseases and even cancer. Therefore, it is necessary to develop accurate and reliable HClO / ClO - Detection methods are of great significance to life science and medical research.

[0006] Hydrazine hydrate (N2H4·H2O) is a colorless, oily liquid with a faint ammoniacal odor that emits fumes and a pungent odor in the air. Its strong nucleophilicity and reducing properties make it an important chemical industry player, widely used as aerospace rocket and jet engine fuel, a dye, an antioxidant, a catalyst, and a polymer crosslinker. It is also used in the synthetic industry, including pharmaceutical and pesticide synthesis, and as a plastic foaming agent. Due to its high toxicity, highly reactive alkalinity, and excellent water solubility, hydrazine hydrate poses a serious threat to the environment and human health. It can enter the human body through inhalation, skin contact, or ingestion, disrupting DNA sequences and damaging the lungs, liver, and central nervous system, causing irreversible damage. Research suggests that the high toxicity of hydrazine hydrate may be related to its easy oxidation into chemically active substances, which can denature proteins, further damage the liver, and even cause death. Summary of the Invention

[0007] To overcome the technical drawbacks of existing instruments, which are expensive and complex to operate, this application provides a new fluorescent probe based on 2,5-dioctyl-3,6-diphenylpyrrolopyrrole dione. By introducing an aldehyde group into 2,5-dioctyl-3,6-diphenylpyrrolopyrrole dione, three fluorescent probes, HG1, DPP-1, and DPP-2, were synthesized through the reaction of the aldehyde group with ethanethiol, diaminomaleonitrile, and malononitrile, enabling the detection of specific substances.

[0008] The present application provides three fluorescent probes HG1, DPP-1 and DPP-2 based on 2,5-dioctyl-3,6-diphenylpyrrolopyrrole dione, which have good fluorescence properties and can achieve simple, efficient and specific detection of specific substances.

[0009] A fluorescent probe based on 2,5-dioctyl-3,6-diphenylpyrrolopyrrole dione, the specific structure of which is as follows:

[0010]

[0011] The synthetic route is as follows:

[0012]

[0013] A fluorescent probe based on 2,5-dioctyl-3,6-diphenylpyrrolopyrrole dione was prepared as follows:

[0014] 1) Weigh 4-cyanobenzaldehyde, aminosulfonic acid, and ethylene glycol and dissolve them in cyclohexane. After reflux reaction for 10-12 hours, cool to room temperature, extract the reaction solution with dichloromethane, wash with saturated sodium bicarbonate solution and water, and remove water from the organic phase with anhydrous magnesium sulfate. Filter and rotary evaporate to obtain a crude product. The crude product is recrystallized from ethanol, filtered, and dried to obtain compound 1;

[0015] 2) Ethyl benzoyl acetate and ethyl chloroacetate were weighed and dissolved in a solvent, anhydrous potassium carbonate and sodium iodide were added, and the mixture was heated to 60-75°C, reacted for 10-14 hours, cooled to room temperature, and filtered. The filtrate was extracted with dichloromethane and washed three times with water. The organic phase was dehydrated with anhydrous magnesium sulfate, filtered, rotary evaporated, and dried to obtain compound 2;

[0016] 3) Weigh ammonium acetate and add it to glacial acetic acid under nitrogen atmosphere. Heat to 65-75°C and stir until completely dissolved. Add compound 2 obtained in step 2) and react for 20-24 hours. Cool to room temperature and pour the reaction solution into ice water. After standing for 15 minutes, filter with suction, wash with water three times, recrystallize with ethanol, filter with suction, and dry to obtain compound 3.

[0017] 4) Weighing sodium metal, adding it to anhydrous tert-amyl alcohol under nitrogen atmosphere, heating to 95-105°C, reacting until the sodium is completely dissolved, then reducing the temperature to 50-60°C, adding compound 1 obtained in step 1) and compound 3 obtained in step 3), heating to 95-105°C, reacting for 10-14 hours, cooling to room temperature, adding methanol and acetic acid, stirring at room temperature for 1-2 hours, filtering, repeatedly washing with methanol and water until the filtrate is colorless, and drying to obtain compound 4;

[0018] 5) Weigh potassium tert-butoxide and compound 4 obtained in step 4), add them to anhydrous N-methylpyrrolidone under nitrogen, heat to 55-65° C., stir until the solution turns red, add n-octane bromide, react for 20-24 hours, cool to room temperature, filter, wash the residue with dichloromethane, combine the filtrate and dichloromethane, wash with concentrated brine and water, dehydrate the organic phase with anhydrous magnesium sulfate, filter, rotary evaporation, column chromatography separation, and dry to obtain compound 5;

[0019] 6) Compound 5 obtained in step 5) was weighed and dissolved in 30 ml of tetrahydrofuran. 0.02 mol / L hydrochloric acid was added and refluxed for 10-12 hours. The mixture was cooled to room temperature, and the reaction solution was extracted with dichloromethane and washed with water three times. The organic layer was dehydrated with anhydrous magnesium sulfate, filtered, and rotary evaporated to obtain a crude product. The crude product was recrystallized from ethanol, filtered, and dried to obtain compound 6;

[0020] 7) Ethanethiol, boron trifluoride etherate, and compound 6 obtained in step 6) were weighed and added to anhydrous dichloromethane under nitrogen protection. The mixture was reacted at 0°C for 6-8 hours, then warmed to room temperature. The reaction solution was rotary evaporated to obtain a crude product, which was separated by column chromatography and dried to obtain compound HG1;

[0021] 8) Weigh diaminomaleonitrile and compound 6 obtained in step 6) and dissolve them in anhydrous ethanol. After reflux reaction for 8-10 hours, cool to room temperature, place in a refrigerator for crystallization overnight, filter, wash with cold ethanol, and dry to obtain compound DPP-1;

[0022] 9) Weigh malononitrile and compound 6 obtained in step 6) and dissolve them in anhydrous ethanol. After reflux reaction for 10-12 hours, cool to room temperature, place in a refrigerator for crystallization overnight, filter, wash with cold ethanol, and dry to obtain compound DPP-2.

[0023] In step 1), the molar ratio of 4-cyanobenzaldehyde to ethylene glycol is 1:5 to 1:8; the molar ratio of 4-cyanobenzaldehyde to aminosulfonic acid is 100:8 to 100:10; the organic phase is first washed with saturated sodium bicarbonate 1 to 2 times, and then washed with water 2 to 3 times; a water separator needs to be installed during the reaction.

[0024] In step 2), the molar ratio of ethyl benzoylacetate to ethyl chloroacetate is 1:1 to 1:2; the molar ratio of ethyl benzoylacetate to anhydrous potassium carbonate is 1:1 to 1:2; the molar ratio of ethyl benzoylacetate to sodium iodide is 5:1 to 5:2; and the solvent is ethylene glycol dimethyl ether and acetone, and the volume ratio of ethylene glycol dimethyl ether to acetone is 1:2 to 1:3.

[0025] In step 3), add 2-3 g of ammonium acetate to every 10 ml of glacial acetic acid.

[0026] In step 4), 0.2 to 0.3 g of metallic sodium is added to every 10 ml of anhydrous tert-amyl alcohol; the molar ratio of compound 3 to metallic sodium is 1:6 to 1:10; the molar ratio of compound 3 to compound 1 is 1:1 to 1:1.5; and the volume ratio of acetic acid to methanol is 1:15 to 1:20.

[0027] In step 5), the molar ratio of compound 4 to potassium tert-butoxide is 1:2 to 1:3; the molar ratio of compound 4 to n-octane bromide is 1:5 to 1:8; the concentrated brine is prepared by adding 1 to 2 g of sodium chloride per 10 ml of pure water; the organic phase is first washed with concentrated brine 4 to 6 times, and then washed with water 4 to 6 times; in column chromatography separation, the n-octane bromide is first washed away with a large amount of petroleum ether, and then eluted with ethyl acetate and petroleum ether, with the volume ratio of ethyl acetate to petroleum ether being 3:10.

[0028] In step 6), the volume ratio of 0.02 mol / L hydrochloric acid to tetrahydrofuran is 1:2 to 1:3.

[0029] In step 7), the molar ratio of compound 6 to ethyl mercaptan is 1:2 to 1:2.5; the molar ratio of boron trifluoride ether to ethyl mercaptan is 1:1 to 1:2; and the column chromatography is eluted with ethyl acetate and petroleum ether, with a volume ratio of ethyl acetate to petroleum ether of 3:10.

[0030] In step 8), the molar ratio of compound 6 to diaminomaleonitrile is 1:1 to 1:2.

[0031] In step 9), the molar ratio of compound 6 to malononitrile is 1:1 to 1:2.

[0032] The fluorescent probe based on 2,5-dioctyl-3,6-diphenylpyrrolopyrrole dione is used in the preparation of mercury ion fluorescent probes, hypochlorite ion fluorescent probes and hydrazine hydrate fluorescent probes.

[0033] Beneficial effects

[0034] The present invention synthesizes three compounds HG1, DPP-1 and DPP-2 based on 2,5-dioctyl-3,6-diphenylpyrrolopyrrole dione.

[0035] Hg 2+ The dithioacetal protecting group in compound HG1 can be selectively removed, converting the dithioacetal in the structure of compound HG1 into an aldehyde group and producing compound 6. The reaction is as follows:

[0036]

[0037] After testing, compound HG1 has the ability to inhibit Hg 2+ Good selectivity, can be applied to Hg 2+ Detection field.

[0038] ClO - It can selectively undergo a nucleophilic addition reaction with -C=N- in the compound DPP-1, followed by a hydrolysis reaction and an elimination reaction to generate an aldehyde and produce compound 6. The reaction is as follows:

[0039]

[0040] After testing, the compound DPP-1 has the ability to inhibit ClO - Has good selectivity and can be applied to ClO - Detection field.

[0041] N2H4 can selectively attack the malononitrile portion of the compound DPP-2 with strong electron-withdrawing ability to generate the corresponding hydrazone. The reaction is as follows:

[0042]

[0043] After testing, the compound DPP-2 has good selectivity for hydrazine hydrate and can be used in the field of hydrazine hydrate detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] (1) Figure 1 is the H NMR spectrum of compound HG1;

[0045] (2) Figure 2 is the mass spectrum of compound HG1;

[0046] (3) Figure 3 This is the fluorescence spectrum of compound HG1's selectivity for metal ion mercury (the horizontal axis is the emission wavelength, and the vertical axis is the fluorescence intensity);

[0047] (4) Figure 4 This is the anti-interference experimental diagram of compound HG1 detecting mercury ions;

[0048] (5) Figure 5 Compound HG1, compound HG1+Hg 2+ , H spectrum comparison of compound 6;

[0049] (6) Figure 6 is the H NMR spectrum of compound DPP-1;

[0050] (7) Figure 7 is the mass spectrum of compound DPP-1;

[0051] (8) Figure 8 This is the fluorescence spectrum of the compound DPP-1's selectivity for hypochlorite (the horizontal axis is the emission wavelength, and the vertical axis is the fluorescence intensity);

[0052] (9) Figure 9 This is the anti-interference experimental diagram of the compound DPP-1 detecting hypochlorite ions;

[0053] (10) Figure 10 Compound DPP-1, compound DPP-1+ClO - , H spectrum comparison of compound 6;

[0054] (11) Figure 11 is the H NMR spectrum of compound DPP-2;

[0055] (12) Figure 12 is the mass spectrum of compound DPP-2;

[0056] (13) Figure 13 This is the fluorescence spectrum of the selectivity of compound DPP-2 for hydrazine hydrate (the horizontal axis is the emission wavelength, and the vertical axis is the fluorescence intensity);

[0057] (14) Figure 14This is a diagram of the anti-interference experiment of compound DPP-2 in detecting hydrazine hydrate;

[0058] (15) Figure 15 This is a high-resolution mass spectrum of compound DPP-2 after adding hydrazine hydrate. DETAILED DESCRIPTION

[0059] In order to better understand the technical solution of the present invention, it is further described in detail below through specific embodiments.

[0060] Example 1

[0061] 1.31 g of 4-cyanobenzaldehyde, 0.097 g of aminosulfonic acid, and 4.96 g of ethylene glycol were dissolved in 60 ml of cyclohexane and refluxed for 12 h. The mixture was then cooled to room temperature and extracted with dichloromethane. The mixture was washed once with 100 ml of saturated sodium bicarbonate solution and twice with 100 ml of water. The organic phase was dehydrated with anhydrous magnesium sulfate, filtered, and rotary evaporated to obtain a crude product. The crude product was recrystallized from ethanol, filtered, and dried to obtain 1.63 g of compound 1 (yield: 93%).

[0062] 1.92 g of ethyl benzoylacetate, 1.22 g of ethyl chloroacetate, 2.07 g of anhydrous potassium carbonate, and 0.60 g of sodium iodide were added to a mixed solvent consisting of 20 ml of ethylene glycol dimethyl ether and 40 ml of acetone, heated to 65°C, reacted for 12 h, cooled to room temperature, filtered, and the filtrate was extracted with dichloromethane and washed three times with water. The organic phase was dehydrated with anhydrous magnesium sulfate, filtered, rotary evaporated, and dried to obtain compound 2;

[0063] 15 g of ammonium acetate was weighed and added to 50 ml of glacial acetic acid under nitrogen atmosphere. The mixture was heated to 65° C. and stirred until completely dissolved. Compound 2 obtained in step 2) was added and allowed to react for 24 h. The mixture was cooled to room temperature and poured into ice water. The reaction mixture was allowed to stand for 15 minutes, filtered, washed with water three times, recrystallized with ethanol, filtered, and dried to obtain 1.50 g of compound 3. The yield was 65% (relative to ethyl benzoyl acetate).

[0064] 1 g of sodium metal was weighed and added to 50 ml of anhydrous tert-amyl alcohol under nitrogen atmosphere. The mixture was heated to 105°C and allowed to react until the sodium was completely dissolved. The temperature was then lowered to 60°C, and 1.05 g of compound 1 obtained in step 1) and 1.16 g of compound 3 obtained in step 3) were added. The mixture was heated to 95°C and allowed to react for 12 h. The mixture was then cooled to room temperature, and 150 ml of methanol and 10 ml of acetic acid were added. The mixture was stirred at room temperature for 2 hours, filtered, and washed repeatedly with methanol and water until the filtrate was colorless. The filtrate was dried to obtain 1.11 g of compound 4, with a yield of 62%.

[0065] 1.12 g of potassium tert-butoxide and 1.74 g of compound 4 were weighed and added to anhydrous N-methylpyrrolidone under nitrogen atmosphere. The mixture was heated to 60°C and stirred until the solution turned red. 5.79 g of octyl bromide was added and the mixture was allowed to react for 24 h. The mixture was then cooled to room temperature and filtered. The residue was washed with dichloromethane. The filtrate and dichloromethane were combined and washed six times with 100 ml of concentrated brine and six times with 100 ml of water. The organic phase was dehydrated with anhydrous magnesium sulfate, filtered, rotary evaporated, separated by column chromatography, and dried to obtain 1.84 g of compound 5 with a yield of 63%.

[0066] 1.43 g of compound 5 obtained in step 5) was weighed and dissolved in 30 ml of tetrahydrofuran. 15 ml of 0.02 mol / L hydrochloric acid was added and refluxed for 12 hours. The reaction mixture was cooled to room temperature and extracted with dichloromethane and washed three times with water. The organic layer was dehydrated with anhydrous magnesium sulfate, filtered, and rotary evaporated to obtain a crude product. The crude product was recrystallized from ethanol, filtered, and dried to obtain 1.28 g of compound 6, with a yield of 95%.

[0067] 0.171 g of ethanethiol, 0.39 g of boron trifluoride etherate, and 0.675 g of compound 6 obtained in step 6) were weighed and added to anhydrous dichloromethane under nitrogen protection. After reacting at 0°C for 8 h, the mixture was warmed to room temperature and the reaction solution was rotary evaporated to obtain a crude product. The crude product was separated by column chromatography and dried to obtain 0.73 g of compound HG1, with a yield of 90%;

[0068] 0.162 g of diaminomaleonitrile and 0.675 g of compound 6 obtained in step 6) were weighed and added to anhydrous ethanol. After reflux for 10 h, the mixture was cooled to room temperature and placed in a refrigerator for crystallization overnight. The mixture was filtered, washed with cold ethanol, and dried to obtain 0.748 g of compound DPP-1, with a yield of 95%.

[0069] 0.099 g of malononitrile and 0.675 g of compound 6 obtained in step 6) were weighed and added to anhydrous ethanol. After reflux for 12 h, the mixture was cooled to room temperature and placed in a refrigerator for crystallization overnight. The mixture was then filtered, washed with cold ethanol, and dried to obtain 0.691 g of compound DPP-2 with a yield of 94%.

[0070] Example 2

[0071] 1.31 g of 4-cyanobenzaldehyde, 0.097 g of aminosulfonic acid, and 4.96 g of ethylene glycol were dissolved in 60 ml of cyclohexane and refluxed for 10 h. The mixture was then cooled to room temperature and extracted with dichloromethane. The mixture was washed once with 100 ml of saturated sodium bicarbonate solution and twice with 100 ml of water. The organic phase was dehydrated with anhydrous magnesium sulfate, filtered, and rotary evaporated to obtain a crude product. The crude product was recrystallized from ethanol, filtered, and dried to obtain 1.56 g of compound 1 (yield: 89%).

[0072] 1.92 g of ethyl benzoylacetate, 1.22 g of ethyl chloroacetate, 2.07 g of anhydrous potassium carbonate, and 0.60 g of sodium iodide were added to a mixed solvent consisting of 20 ml of ethylene glycol dimethyl ether and 40 ml of acetone, heated to 65°C, reacted for 12 h, cooled to room temperature, filtered, and the filtrate was extracted with dichloromethane and washed three times with water. The organic phase was dehydrated with anhydrous magnesium sulfate, filtered, rotary evaporated, and dried to obtain compound 2;

[0073] 15 g of ammonium acetate was weighed and added to 50 ml of glacial acetic acid under nitrogen atmosphere. The mixture was heated to 75°C and stirred until completely dissolved. Compound 2 obtained in step 2) was added and allowed to react for 20 h. The mixture was then cooled to room temperature and poured into ice water. The reaction mixture was allowed to stand for 15 minutes, filtered, washed with water three times, recrystallized with ethanol, filtered, and dried to obtain 1.386 g of compound 3. The yield was 60% (relative to ethyl benzoyl acetate).

[0074] 1 g of sodium metal was weighed and added to 50 ml of anhydrous tert-amyl alcohol under nitrogen atmosphere. The mixture was heated to 105°C and allowed to react until the sodium was completely dissolved. The temperature was then lowered to 60°C, and 1.05 g of compound 1 obtained in step 1) and 1.16 g of compound 3 obtained in step 3) were added. The mixture was heated to 95°C and allowed to react for 10 h. The mixture was then cooled to room temperature, and 150 ml of methanol and 10 ml of acetic acid were added. The mixture was stirred at room temperature for 2 hours, filtered, and washed repeatedly with methanol and water until the filtrate was colorless. The filtrate was dried to obtain 1.04 g of compound 4, with a yield of 58%.

[0075] 1.12 g of potassium tert-butoxide and 1.74 g of compound 4 were weighed and added to anhydrous N-methylpyrrolidone under nitrogen atmosphere. The mixture was heated to 65°C and stirred until the solution turned red. 5.79 g of octyl bromide was added and the mixture was allowed to react for 20 h. The mixture was then cooled to room temperature and filtered. The residue was washed with dichloromethane. The filtrate and dichloromethane were combined and washed six times with 100 ml of concentrated brine and six times with 100 ml of water. The organic phase was dehydrated with anhydrous magnesium sulfate, filtered, rotary evaporated, separated by column chromatography, and dried to obtain 1.63 g of compound 5 with a yield of 56%.

[0076] 1.43 g of compound 5 obtained in step 5) was weighed and dissolved in 30 ml of tetrahydrofuran. 10 ml of 0.02 mol / L hydrochloric acid was added and refluxed for 12 hours. The reaction mixture was cooled to room temperature and extracted with dichloromethane and washed three times with water. The organic layer was dehydrated with anhydrous magnesium sulfate, filtered, and rotary evaporated to obtain a crude product. The crude product was recrystallized from ethanol, filtered, and dried to obtain 1.18 g of compound 6, with a yield of 88%.

[0077] 0.171 g of ethanethiol, 0.39 g of boron trifluoride etherate, and 0.675 g of compound 6 obtained in step 6) were weighed and added to anhydrous dichloromethane under nitrogen protection. After reacting at 0°C for 6 h, the mixture was warmed to room temperature and the reaction solution was rotary evaporated to obtain a crude product. The crude product was separated by column chromatography and dried to obtain 0.70 g of compound HG1, with a yield of 87%.

[0078] 0.162 g of diaminomaleonitrile and 0.675 g of compound 6 obtained in step 6) were weighed and added to anhydrous ethanol. After reflux for 8 h, the mixture was cooled to room temperature and placed in a refrigerator for crystallization overnight. The mixture was then filtered, washed with cold ethanol, and dried to obtain 0.717 g of compound DPP-1, with a yield of 91%.

[0079] 0.099 g of malononitrile and 0.675 g of compound 6 obtained in step 6) were weighed and added to anhydrous ethanol. After reflux for 10 h, the mixture was cooled to room temperature and placed in a refrigerator for crystallization overnight. The mixture was then filtered, washed with cold ethanol, and dried to obtain 0.662 g of compound DPP-2 with a yield of 90%.

[0080] Example 3

[0081] 1.31 g of 4-cyanobenzaldehyde, 0.078 g of aminosulfonic acid, and 4.96 g of ethylene glycol were dissolved in 60 ml of cyclohexane and refluxed for 12 h. The mixture was then cooled to room temperature and extracted with dichloromethane. The mixture was washed once with 100 ml of saturated sodium bicarbonate solution and twice with 100 ml of water. The organic phase was dehydrated with anhydrous magnesium sulfate, filtered, and rotary evaporated to obtain a crude product. The crude product was recrystallized from ethanol, filtered, and dried to obtain 1.54 g of compound 1 (yield: 88%).

[0082] 1.92 g of ethyl benzoylacetate, 1.22 g of ethyl chloroacetate, 2.07 g of anhydrous potassium carbonate, and 0.60 g of sodium iodide were added to a mixed solvent consisting of 20 ml of ethylene glycol dimethyl ether and 40 ml of acetone, heated to 65°C, reacted for 12 h, cooled to room temperature, filtered, and the filtrate was extracted with dichloromethane and washed three times with water. The organic phase was dehydrated with anhydrous magnesium sulfate, filtered, rotary evaporated, and dried to obtain compound 2;

[0083] 10 g of ammonium acetate was weighed and added to 50 ml of glacial acetic acid under nitrogen atmosphere. The mixture was heated to 65° C. and stirred until completely dissolved. Compound 2 obtained in step 2) was added and allowed to react for 24 h. The mixture was cooled to room temperature and poured into ice water. The reaction solution was allowed to stand for 15 minutes, filtered, washed with water three times, recrystallized with ethanol, filtered, and dried to obtain 1.04 g of compound 3. The yield was 45% (relative to ethyl benzoyl acetate).

[0084] 1.5 g of sodium metal was weighed and added to 50 ml of anhydrous tert-amyl alcohol under nitrogen atmosphere. The mixture was heated to 105° C. and allowed to react until the sodium was completely dissolved. The temperature was then lowered to 60° C., and 1.05 g of compound 1 obtained in step 1) and 1.16 g of compound 3 obtained in step 3) were added. The mixture was heated to 95° C. and allowed to react for 12 h. The mixture was then cooled to room temperature, and 150 ml of methanol and 10 ml of acetic acid were added. The mixture was stirred at room temperature for 2 hours, filtered, and washed repeatedly with methanol and water until the filtrate was colorless. The filtrate was dried to obtain 0.92 g of compound 4, with a yield of 51%.

[0085] 1.12 g of potassium tert-butoxide and 1.74 g of compound 4 were weighed and added to anhydrous N-methylpyrrolidone under nitrogen atmosphere. The mixture was heated to 60°C and stirred until the solution turned red. 4.82 g of octyl bromide was added and the mixture was allowed to react for 24 h. The mixture was then cooled to room temperature and filtered. The residue was washed with dichloromethane. The filtrate and dichloromethane were combined and washed six times with 100 ml of concentrated brine and six times with 100 ml of water. The organic phase was dehydrated with anhydrous magnesium sulfate, filtered, rotary evaporated, separated by column chromatography, and dried to obtain 1.75 g of compound 5 with a yield of 60%.

[0086] 1.43 g of compound 5 obtained in step 5) was weighed and dissolved in 30 ml of tetrahydrofuran. 10 ml of 0.02 mol / L hydrochloric acid was added and refluxed for 12 hours. The mixture was cooled to room temperature and extracted with dichloromethane and washed three times with water. The organic layer was dehydrated with anhydrous magnesium sulfate, filtered, and rotary evaporated to obtain a crude product. The crude product was recrystallized from ethanol, filtered, and dried to obtain 1.22 g of compound 6, with a yield of 90%.

[0087] 0.155 g of ethanethiol, 0.39 g of boron trifluoride etherate, and 0.675 g of compound 6 obtained in step 6) were weighed and added to anhydrous dichloromethane under nitrogen protection. After reacting at 0°C for 8 h, the mixture was warmed to room temperature and the reaction solution was rotary evaporated to obtain a crude product. The crude product was separated by column chromatography and dried to obtain 0.65 g of compound HG1, with a yield of 81%;

[0088] 0.135 g of diaminomaleonitrile and 0.675 g of compound 6 obtained in step 6) were weighed and added to anhydrous ethanol. After reflux for 10 h, the mixture was cooled to room temperature and placed in a refrigerator for crystallization overnight. The mixture was then filtered, washed with cold ethanol, and dried to obtain 0.701 g of compound DPP-1, with a yield of 89%.

[0089] 0.082 g of malononitrile and 0.675 g of compound 6 obtained in step 6) were weighed and added to anhydrous ethanol. After reflux for 12 h, the mixture was cooled to room temperature and placed in a refrigerator for crystallization overnight. The mixture was then filtered, washed with cold ethanol, and dried to obtain 0.647 g of compound DPP-2 with a yield of 88%.

[0090] Example 4

[0091] 1.31 g of 4-cyanobenzaldehyde, 0.097 g of aminosulfonic acid, and 3.10 g of ethylene glycol were dissolved in 60 ml of cyclohexane and refluxed for 12 h. The mixture was then cooled to room temperature and extracted with dichloromethane. The mixture was washed once with 100 ml of saturated sodium bicarbonate solution and twice with 100 ml of water. The organic phase was dehydrated with anhydrous magnesium sulfate, filtered, and rotary evaporated to obtain a crude product. The crude product was recrystallized from ethanol, filtered, and dried to obtain 1.47 g of compound 1 (yield: 84%).

[0092] 1.92 g of ethyl benzoylacetate, 1.22 g of ethyl chloroacetate, 1.38 g of anhydrous potassium carbonate, and 0.60 g of sodium iodide were weighed and added to a mixed solvent consisting of 20 ml of ethylene glycol dimethyl ether and 40 ml of acetone. The mixture was heated to 65°C and reacted for 12 h. The mixture was then cooled to room temperature and filtered. The filtrate was extracted with dichloromethane and washed three times with water. The organic phase was dehydrated with anhydrous magnesium sulfate, filtered, rotary evaporated, and dried to obtain compound 2.

[0093] 15 g of ammonium acetate was weighed and added to 50 ml of glacial acetic acid under nitrogen atmosphere. The mixture was heated to 65° C. and stirred until completely dissolved. Compound 2 obtained in step 2) was added and allowed to react for 24 h. The mixture was cooled to room temperature and poured into ice water. The reaction mixture was allowed to stand for 15 minutes, filtered, washed with water three times, recrystallized with ethanol, filtered, and dried to obtain 1.32 g of compound 3. The yield was 57% (relative to ethyl benzoyl acetate).

[0094] 1 g of sodium metal was weighed and added to 50 ml of anhydrous tert-amyl alcohol under nitrogen atmosphere. The mixture was heated to 105°C and allowed to react until the sodium was completely dissolved. The temperature was then lowered to 60°C, and 1.31 g of compound 1 obtained in step 1) and 1.16 g of compound 3 obtained in step 3) were added. The mixture was heated to 95°C and allowed to react for 12 h. The mixture was then cooled to room temperature, and 150 ml of methanol and 10 ml of acetic acid were added. The mixture was stirred at room temperature for 2 hours, filtered, and washed repeatedly with methanol and water until the filtrate was colorless. The filtrate was dried to obtain 1.08 g of compound 4, with a yield of 60%.

[0095] 1.12 g of potassium tert-butoxide and 1.74 g of compound 4 were weighed and added to anhydrous N-methylpyrrolidone under nitrogen atmosphere. The mixture was heated to 60°C and stirred until the solution turned red. 7.72 g of octyl bromide was added and the mixture was allowed to react for 24 h. The mixture was then cooled to room temperature and filtered. The residue was washed with dichloromethane. The filtrate and dichloromethane were combined and washed six times with 100 ml of brine and six times with 100 ml of water. The organic phase was dehydrated with anhydrous magnesium sulfate, filtered, rotary evaporated, separated by column chromatography, and dried to obtain 1.81 g of compound 5 with a yield of 62%.

[0096] 1.43 g of compound 5 obtained in step 5) was weighed and dissolved in 30 ml of tetrahydrofuran. 15 ml of 0.02 mol / L hydrochloric acid was added and refluxed for 12 hours. The reaction mixture was cooled to room temperature and extracted with dichloromethane and washed three times with water. The organic layer was dehydrated with anhydrous magnesium sulfate, filtered, and rotary evaporated to obtain a crude product. The crude product was recrystallized from ethanol, filtered, and dried to obtain 1.28 g of compound 6, with a yield of 95%.

[0097] 0.171 g of ethanethiol, 0.53 g of boron trifluoride etherate, and 0.675 g of compound 6 obtained in step 6) were weighed and added to anhydrous dichloromethane under nitrogen protection. After reacting at 0°C for 8 h, the mixture was warmed to room temperature and the reaction solution was rotary evaporated to obtain a crude product. The crude product was separated by column chromatography and dried to obtain 0.73 g of compound HG1, with a yield of 90%;

[0098] 0.202 g of diaminomaleonitrile and 0.675 g of compound 6 obtained in step 6) were weighed and added to anhydrous ethanol. After reflux for 8 h, the mixture was cooled to room temperature and placed in a refrigerator for crystallization overnight. The mixture was then filtered, washed with cold ethanol, and dried to obtain 0.740 g of compound DPP-1, with a yield of 94%.

[0099] 0.165 g of malononitrile and 0.675 g of compound 6 obtained in step 6) were weighed and added to anhydrous ethanol. After reflux for 10 h, the mixture was cooled to room temperature and placed in a refrigerator for crystallization overnight. The mixture was then filtered, washed with cold ethanol, and dried to obtain 0.676 g of compound DPP-2 with a yield of 92%.

[0100] Example 5

[0101] 1.31 g of 4-cyanobenzaldehyde, 0.078 g of aminosulfonic acid, and 3.10 g of ethylene glycol were dissolved in 60 ml of cyclohexane and refluxed for 12 h. The mixture was then cooled to room temperature and extracted with dichloromethane. The mixture was washed once with 100 ml of saturated sodium bicarbonate solution and twice with 100 ml of water. The organic phase was dehydrated with anhydrous magnesium sulfate, filtered, and rotary evaporated to obtain a crude product. The crude product was recrystallized from ethanol, filtered, and dried to obtain 1.38 g of compound 1 (yield: 79%).

[0102] 1.92 g of ethyl benzoylacetate, 1.22 g of ethyl chloroacetate, 2.07 g of anhydrous potassium carbonate, and 0.30 g of sodium iodide were weighed and added to a mixed solvent consisting of 20 ml of ethylene glycol dimethyl ether and 40 ml of acetone. The mixture was heated to 65°C and reacted for 12 h. The mixture was then cooled to room temperature and filtered. The filtrate was extracted with dichloromethane and washed three times with water. The organic phase was dehydrated with anhydrous magnesium sulfate, filtered, rotary evaporated, and dried to obtain compound 2.

[0103] 15 g of ammonium acetate was weighed and added to 50 ml of glacial acetic acid under nitrogen atmosphere. The mixture was heated to 65° C. and stirred until completely dissolved. Compound 2 obtained in step 2) was added and allowed to react for 24 h. The mixture was cooled to room temperature and poured into ice water. The reaction mixture was allowed to stand for 15 minutes, filtered, washed with water three times, recrystallized with ethanol, filtered, and dried to obtain 1.39 g of compound 3. The yield was 60% (relative to ethyl benzoyl acetate).

[0104] 1 g of sodium metal was weighed and added to 50 ml of anhydrous tert-amyl alcohol under nitrogen atmosphere. The mixture was heated to 105°C and allowed to react until the sodium was completely dissolved. The temperature was then lowered to 60°C, and 1.05 g of compound 1 obtained in step 1) and 1.16 g of compound 3 obtained in step 3) were added. The mixture was heated to 95°C and allowed to react for 12 h. The mixture was then cooled to room temperature, and 150 ml of methanol and 15 ml of acetic acid were added. The mixture was stirred at room temperature for 2 hours, filtered, and washed repeatedly with methanol and water until the filtrate was colorless. The filtrate was dried to obtain 1.11 g of compound 4 with a yield of 62%.

[0105] 1.12 g of potassium tert-butoxide and 1.74 g of compound 4 were weighed and added to anhydrous N-methylpyrrolidone under nitrogen atmosphere. The mixture was heated to 60°C and stirred until the solution turned red. 5.79 g of octyl bromide was added and the mixture was allowed to react for 24 h. The mixture was then cooled to room temperature and filtered. The residue was washed with dichloromethane. The filtrate and dichloromethane were combined and washed six times with 100 ml of concentrated brine and six times with 100 ml of water. The organic phase was dehydrated with anhydrous magnesium sulfate, filtered, rotary evaporated, separated by column chromatography, and dried to obtain 1.84 g of compound 5 with a yield of 63%.

[0106] 1.43 g of compound 5 obtained in step 5) was weighed and dissolved in 30 ml of tetrahydrofuran. 15 ml of 0.02 mol / L hydrochloric acid was added and refluxed for 12 hours. The reaction mixture was cooled to room temperature and extracted with dichloromethane and washed three times with water. The organic layer was dehydrated with anhydrous magnesium sulfate, filtered, and rotary evaporated to obtain a crude product. The crude product was recrystallized from ethanol, filtered, and dried to obtain 1.28 g of compound 6, with a yield of 95%.

[0107] 0.155 g of ethanethiol, 0.26 g of boron trifluoride etherate, and 0.675 g of compound 6 obtained in step 6) were weighed and added to anhydrous dichloromethane under nitrogen protection. After reacting at 0°C for 8 h, the mixture was warmed to room temperature and the reaction solution was rotary evaporated to obtain a crude product. The crude product was separated by column chromatography and dried to obtain 0.65 g of compound HG1, with a yield of 81%;

[0108] 0.27 g of diaminomaleonitrile and 0.675 g of compound 6 obtained in step 6) were weighed and added to anhydrous ethanol. After reflux for 10 h, the mixture was cooled to room temperature and placed in a refrigerator for crystallization overnight. The mixture was then filtered, washed with cold ethanol, and dried to obtain 0.740 g of compound DPP-1, with a yield of 94%.

[0109] 0.165 g of malononitrile and 0.675 g of compound 6 obtained in step 6) were weighed and added to anhydrous ethanol. After reflux for 12 h, the mixture was cooled to room temperature and placed in a refrigerator for crystallization overnight. The mixture was then filtered, washed with cold ethanol, and dried to obtain 0.691 g of compound DPP-2 with a yield of 94%.

[0110] HG1 compound HNMR spectrum (attached Figure 1 ) and mass spectrometry analysis (attached Figure 2 ):

[0111] Table 1 was obtained by analyzing the structural formula and H NMR spectrum of compound HG1. This compound has 15 kinds of hydrogen. The signal peak appearing near 0.79ppm is the signal peak of proton 1, and its peak area is 6.00; the signal peak appearing in the range of 1.07-1.21ppm is the signal peak of protons 2, 3, 4, 5, 6 and 15, and its peak area is 26.15; the signal peak appearing near 1.36ppm is the signal peak of proton 7, and its peak area is 4.05; the signal peak appearing near 3.67ppm is the signal peak of proton 8, and its peak area is 4.07; the signal peak appearing near 7.56ppm is the signal peak of proton 1, and its peak area is 2. The peak is the signal peak of proton 9, and its peak area is 2.99; the signal peak appearing near 7.62ppm is the signal peak of proton 10, and its peak area is 2.101; the signal peak appearing near 7.81ppm is the signal peak of protons 11 and 12, and its peak area is 4.05; the signal peak appearing near 5.28ppm is the signal peak of proton 13, and its peak area is 1.00; the signal peak appearing near 2.50-2.65ppm is the signal peak of proton 14, and its peak area is 4.05. It can be seen from this that the H NMR spectrum of the compound is in good agreement with the structure of the compound.

[0112] Compound structure C 39 H 54 The molecular weight of N2O2S2 was calculated to be 646.3627 and the molecular weight was tested to be 647.3693 (C 39 H 54 N2O2S2) + From this, it can be concluded that the mass spectrum of the compound is consistent with the expected molecular weight.

[0113] Table 1 Compound HQ1 1 Chemical shifts and peak assignments of HNMR.

[0114] Table 1

[0115]

[0116] DPP-1 compound H NMR spectrum (attached Figure 5 ) and mass spectrometry analysis (attached Figure 6 ):

[0117] Table 2 was obtained by analyzing the structural formula and H NMR spectrum of compound DPP-1. This compound has 14 kinds of hydrogen. Among them, the signal peak appearing near 0.80ppm is the signal peak of proton 1, and its peak area is 6.27; the signal peak appearing in the range of 1.08-1.21ppm is the signal peak of proton 2, 3, 4, 5 and 6, and its peak area is 20.40; the signal peak appearing near 1.38ppm is the signal peak of proton 7, and its peak area is 4.09; the signal peak appearing near 3.72ppm is the signal peak of proton 8, and its peak area is 4.06; the signal peak appearing near 7.59ppm is the signal peak of proton 9, and its peak area is The peak area of ​​the protons is 3.07; the peak at 7.82 ppm is the peak area of ​​proton 10, and its peak area is 2.10; the peak at 7.92 ppm is the peak area of ​​proton 11, and its peak area is 2.05; the peak at 8.23 ​​ppm is the peak area of ​​proton 12, and its peak area is 1.92; the peak at 8.34 ppm is the peak area of ​​proton 13, and its peak area is 1.00; the peak at 8.17 ppm is the peak area of ​​proton 14, and its peak area is 1.90. It can be seen that the H NMR spectrum of the compound is consistent with the structure of the compound.

[0118] Compound structure C 39 H 46 The molecular weight of N6O2 was calculated to be 630.3682 and the molecular weight was tested to be 631.3747 (C 39 H 46 N6O2) + From this, it can be concluded that the mass spectrum of the compound is consistent with the expected molecular weight.

[0119] Table 2 Compound DPP-1 1 Chemical shifts and peak assignments of HNMR.

[0120] Table 2

[0121]

[0122] DPP-2 compound H NMR spectrum (attached Figure 9 ) and mass spectrometry analysis (attached Figure 10 ):

[0123] Table 3 was obtained by analyzing the structural formula and H NMR spectrum of compound DPP-2. This compound has 13 hydrogen species. The signal peak appearing near 0.81ppm is the signal peak of proton 1, and its peak area is 6.08; the signal peak appearing in the range of 1.09-1.23ppm is the signal peak of protons 2, 3, 4, 5 and 6, and its peak area is 20.27; the signal peak appearing near 1.39ppm is the signal peak of proton 7, and its peak area is 4.04; the signal peak appearing near 3.73ppm is the signal peak of proton 8, and its peak area is 4.09; the signal peak appearing near 7.60ppm is the signal peak of proton 1, and its peak area is 4.09; The signal peak at 7.83ppm is the signal peak of proton 10, and its peak area is 2.05; the signal peak at 8.06ppm is the signal peak of proton 11, and its peak area is 2.01; the signal peak at 8.12ppm is the signal peak of proton 12, and its peak area is 2.03; the signal peak at 8.64ppm is the signal peak of proton 13, and its peak area is 1.00. It can be seen that the H NMR spectrum of the compound is consistent with the structure of the compound.

[0124] Compound structure C 38 H 44 The molecular weight of N4O2 was calculated to be 588.3464 and the molecular weight was tested to be 589.3535 (C 38 H 44 N4O2) + From this, it can be concluded that the mass spectrum of the compound is consistent with the expected molecular weight.

[0125] Table 3 Compound DPP-2 1 Chemical shifts and peak assignments of HNMR.

[0126] Table 3

[0127]

[0128] Compound HG1 prepared in Example 1 was dissolved and diluted in tetrahydrofuran: water (1 / 1, v / v) to prepare 1.0×10 -5 mol / L sample solution. The fluorescence excitation wavelength of the compound was measured using an F-700 fluorescence spectrophotometer, and the fluorescence spectrum of the compound was measured. Equivalent amounts of different metal ions Hg 2+ 、Ag + 、Ba + 、Cd + 、Co + 、Cu 2+ 、Fe 2+ 、Fe 3+ , K+ 、Na + 、Ni 2+ , Pb 2+ 、Sr 2+ 、Zn 2+ Determine the fluorescence emission spectrum of the fluorescent probe in the presence of various metal ions (such as the attached Figure 3 As shown), the probe emits yellow fluorescence, and Hg 2+ After that, the fluorescence quickly turned orange and the fluorescence intensity weakened. However, the fluorescence intensity did not change much after adding other ions, and no fluorescence red shift occurred. This result shows that the probe HG1 can effectively inhibit Hg 2+ It has a good identification function.

[0129] The compound DPP-1 prepared in Example 1 was dissolved and diluted in tetrahydrofuran: water (1 / 1, v / v) to prepare 1.0X 10 -5 mol / L sample solution. The fluorescence excitation wavelength of the compound was measured using an F-700 fluorescence spectrophotometer, and the fluorescence spectrum of the compound was measured. Equivalent amounts of different ions ClO were added to the solution. - 、F - Br - 、Cl - 、NO3 - 、HSO3 - 、ACO - 、HCO3 - 、CO3 2- 、S 2- 、SO4 2- , GSH, Cys and H2O2 were measured in the presence of each ion, and the fluorescence emission spectrum of the fluorescent probe (such as the attached Figure 7 As shown), the probe emits red fluorescence, and ClO - After that, the fluorescence quickly turned orange and the fluorescence intensity became stronger. However, the fluorescence intensity did not change much after adding other ions, and no fluorescence blue shift occurred. This result shows that the probe DPP-1 has a strong effect on ClO - It has a good identification function.

[0130] The compound DPP-2 prepared in Example 1 was dissolved and diluted with ethanol: water (2 / 1, v / v) to prepare 1.0×10 -5 mol / L sample solution. The fluorescence excitation wavelength of the compound was measured using an F-700 fluorescence spectrophotometer, and the fluorescence spectrum of the compound was measured. Equivalent amounts of N2H4, Mg 2+ 、Cu 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ , Ca 2+ 、Ba 2+Cr 3+ , ClO - 、F - Br - 、Cl - 、NO3 - 、HSO3 - 、ACO - 、HCO3 - 、CO3 2- 、S 2- 、SO4 2- , GSH, Cys and H2O2 were measured in the presence of each ion, and the fluorescence emission spectrum of the fluorescent probe (such as the attached Figure 11 The probe emits red fluorescence. After adding hydrazine hydrate, the fluorescence quickly turns yellow and the fluorescence intensity becomes stronger. However, when other ions are added, the fluorescence intensity does not change much and no fluorescence blue shift occurs. This result shows that the probe DPP-2 has a good recognition effect on hydrazine hydrate.

[0131] In the presence of other ions, the fluorescent probe HG1 selectively recognizes mercury ions, i.e., ion competition experiments (e.g., Figure 4 The compound HG1 prepared in Example 1 was dissolved and diluted in tetrahydrofuran: water (1 / 1, v / v) to prepare 1.0X 10 -5 mol / L sample solution. Equivalent amounts of different ions (Ag + 、Ba + 、Cd + 、Co + 、Cu 2 + 、Fe 2+ 、Fe 3+ , K + 、Na + 、Ni 2+ , Pb 2+ 、Sr 2+ 、Zn 2+ ), the fluorescence intensity of probe HG1 did not change significantly, while the addition of an equivalent amount of Hg 2+ After the fluorescence intensity increased significantly, it proved that the probe HG1 could still detect Hg even in the presence of other ions. 2+ Has higher selectivity.

[0132] In the presence of other ions, the fluorescent probe DPP-1 selectively recognizes hypochlorite ions, i.e., ion competition experiments (e.g., Figure 8 The compound DPP-1 prepared in Example 1 was dissolved and diluted in tetrahydrofuran: water (1 / 1, v / v) to prepare 1.0X 10 -5mol / L sample solution. Equivalent amounts of different ions (F - Br - 、Cl - 、NO3 - 、HSO3 - 、ACO - 、HCO3 - 、CO3 2- 、S 2- 、SO4 2- , GSH, Cys, H2O2), the fluorescence intensity of the probe DPP-1 did not change significantly, while the addition of an equivalent amount of ClO - After the addition of ions, the fluorescence intensity increased significantly, which proved that the probe HG1 could still detect ClO even in the presence of other ions. - Has higher selectivity.

[0133] In the presence of other ions, the fluorescent probe DPP-2 selectively recognizes mercury ions, i.e., ion competition experiments (e.g., Figure 12 The compound DPP-2 prepared in Example 1 was dissolved and diluted with ethanol: water (2 / 1, v / v) to prepare 1.0X 10 -5 mol / L sample solution. Equivalent amounts of different ions (Mg 2+ 、Cu 2+ 、Fe 2+ 、Fe 3 + 、Al 3+ , Ca 2+ 、Ba 2+ Cr 3+ , ClO - 、F - Br - 、Cl - 、NO3 - 、HSO3 - 、ACO - 、HCO3 - 、CO3 2- 、S 2- 、SO4 2- , GSH, Cys, H2O2), the fluorescence intensity of probe DPP-2 did not change significantly, while after adding an equivalent amount of N2H4, the fluorescence intensity was significantly enhanced, which proved that probe HG1 still had high selectivity for N2H4 even in the presence of other ions.

[0134] The probe HG1 was compared with Hg 2+ 1HNMR spectra before and after the reaction (see attached Figure 5 Probe HG1 and Hg2+ After the reaction, the signal peaks attributed to hydrogen 13 and hydrogen 14 disappeared, and a new peak attributed to aldehyde hydrogen was observed. 2+ The H NMR spectrum after the reaction is consistent with that of compound 6. 2+ Under the promotion of , the thioacetal group is eliminated to produce the corresponding aldehyde group.

[0135] The probe DPP-1 was compared with ClO - Before and after reaction 1 HNMR spectrum (see attached Figure 10 Probe DPP-1 and ClO - After the reaction, the signal peaks attributed to hydrogen 13 and hydrogen 14 disappeared, and a new peak attributed to aldehyde hydrogen was observed. - The H NMR spectrum after the reaction is consistent with that of compound 6. The results show that in ClO - Under the promotion of , the maleonitrile part of the probe DPP-1 is eliminated, thereby generating the corresponding aldehyde group.

[0136] The reaction mixture of probe DPP-2 and excess hydrazine hydrate was detected by high-resolution mass spectrometry (see attached). Figure 15 As shown in Figure 2, the molecular ion peak of probe DPP-2 disappears, and a new molecular ion peak appears at m / z = 555.3691. This peak can be attributed to the hydrazone generated by the reaction, corresponding to the structure C of the hydrazone. 35 H 46 N4O2, its molecular weight is calculated to be 554.3621.

Claims

1. A fluorescent probe based on 2,5-dioctyl-3,6-diphenylpyrrolopyrrole diketo, characterized in that its specific structure is as follows: or .

2. The method for preparing a fluorescent probe based on 2,5-dioctyl-3,6-diphenylpyrrolopyrrole diketo as claimed in claim 1, wherein: The synthetic route is as follows: 。 3. The method for preparing a fluorescent probe based on 2,5-dioctyl-3,6-diphenylpyrrolopyrrole diketo as claimed in claim 2, characterized in that: Here are the steps: 1) Sodium metal was weighed and added to anhydrous tert-amyl alcohol under nitrogen atmosphere. The mixture was heated to 95-105°C and allowed to react until the sodium was completely dissolved. The temperature was then lowered to 50-60°C, compound 1 and compound 3 were added, the mixture was heated to 95-105°C, and the reaction was continued for 10-14 hours. The mixture was then cooled to room temperature, methanol and acetic acid were added, and the mixture was stirred at room temperature for 1-2 hours. The mixture was then filtered and washed repeatedly with methanol and water until the filtrate was colorless. The filtrate was then dried to obtain compound 4; 2) Weighing potassium tert-butoxide and compound 4 obtained in step 1) are added to anhydrous N-methylpyrrolidone under nitrogen atmosphere, heated to 55-65°C, stirred until the solution turns red, and then added with n-octyl bromide. After reacting for 20-24 hours, the mixture is cooled to room temperature and filtered. The filter residue is washed with dichloromethane. The filtrate and dichloromethane are combined and washed with concentrated brine and water. The organic phase is dehydrated with anhydrous magnesium sulfate, filtered, rotary evaporated, separated by column chromatography, and dried to obtain compound 5; 3) Compound 5 obtained in step 2) was weighed and dissolved in 30 ml of tetrahydrofuran. 0.02 mol / L hydrochloric acid was added and refluxed for 10-12 hours. The mixture was cooled to room temperature, and the reaction solution was extracted with dichloromethane and washed with water three times. The organic layer was dehydrated with anhydrous magnesium sulfate, filtered, and rotary evaporated to obtain a crude product. The crude product was recrystallized from ethanol, filtered, and dried to obtain compound 6. 4) Weighing diaminomaleonitrile and compound 6 obtained in step 3) were dissolved in anhydrous ethanol, refluxed for 8-10 hours, cooled to room temperature, placed in a refrigerator for crystallization overnight, filtered, washed with cold ethanol, and dried to obtain compound DPP-1; 5) Weigh malononitrile and compound 6 obtained in step 3) and dissolve them in anhydrous ethanol. After reflux reaction for 10-12 hours, cool to room temperature, place in a refrigerator for crystallization overnight, filter, wash with cold ethanol, and dry to obtain compound DPP-2.

4. The method for preparing a fluorescent probe based on 2,5-dioctyl-3,6-diphenylpyrrolopyrrole diketo as claimed in claim 3, wherein: In step 1), 0.2-0.3 g of metallic sodium is added to every 10 ml of anhydrous tert-amyl alcohol; the molar ratio of compound 3 to metallic sodium is 1:6-1:10; the molar ratio of compound 3 to compound 1 is 1:1-1:1.5; and the volume ratio of acetic acid to methanol is 1:15-1:

20.

5. The method for preparing a fluorescent probe based on 2,5-dioctyl-3,6-diphenylpyrrolopyrrole diketo as claimed in claim 3, wherein: In step 2), the molar ratio of compound 4 to potassium tert-butoxide is 1:2 to 1:3; the molar ratio of compound 4 to n-octane bromide is 1:5 to 1:8; concentrated brine is prepared by adding 1 to 2 g of sodium chloride per 10 ml of water, and the organic phase is first washed with concentrated brine 4 to 6 times and then with pure water 4 to 6 times; in column chromatography separation, n-octane bromide is first washed away with a large amount of petroleum ether, and then eluted with ethyl acetate and petroleum ether, with the volume ratio of ethyl acetate to petroleum ether being 3:

10.

6. The method for preparing a fluorescent probe based on 2,5-dioctyl-3,6-diphenylpyrrolopyrrole diketo as claimed in claim 3, wherein: In step 3), the volume ratio of 0.02 mol / L hydrochloric acid to tetrahydrofuran is 1:2 to 1:

3.

7. The method for preparing a fluorescent probe based on 2,5-dioctyl-3,6-diphenylpyrrolopyrrole diketo as claimed in claim 3, wherein: In step 4), the molar ratio of compound 6 to diaminomaleonitrile is 1:1 to 1:

2.

8. The method for preparing a fluorescent probe based on 2,5-dioctyl-3,6-diphenylpyrrolopyrrole diketo as claimed in claim 3, wherein: In step 5), the molar ratio of compound 6 to malononitrile is 1:1 to 1:

2.

9. The use of a fluorescent probe based on 2,5-dioctyl-3,6-diphenylpyrrolopyrrole diketo as claimed in claim 1, characterized in that: The DPP-1 is a fluorescent probe for preparing hypochlorite ions; and the DPP-2 is a fluorescent probe for preparing hydrazine hydrate.

Images