A class of polypyridine ruthenium complexes and their applications

Abstract

The present invention discloses a type of polypyridine ruthenium complex, wherein the cation of the compound is the polypyridine ruthenium complex of the present invention, which can efficiently and selectively detect CO3 in an aqueous system. 2‑ In addition, due to its different ligand structure, it also has the ‑ , ClO ‑ 、HCO3 ‑ Specific detection is a volunteer method. The present invention can identify the above-mentioned ions by colorimetry. The method is simple, environmentally friendly, rapid in detection and with stable signals, and is suitable for environmental monitoring, food safety and biomedical testing. The complex of the present invention has an aggregation-induced luminescence effect and can be used in the preparation of cell diagnosis and treatment imaging reagents. In the anti-tumor activity test, it was found that the polypyridine ruthenium complex of the present invention has the potential to prepare anti-tumor drugs.

Description

[0001] Technical Essentials

[0002] The present invention relates to a class of polypyridyl ruthenium complexes and their applications in detecting carbonate, bisulfite, bicarbonate, hypochlorite, preparing cell diagnosis and treatment imaging reagents, and preparing anti-tumor drugs, and belongs to the field of environmental and biomedical detection. Background Art

[0003] Transition metal complexes are considered to have the potential to serve as phosphorescent sensors for metal ions, anions, and biomolecules due to their favorable photophysical and chemical properties, including large Stokes shifts, long luminescence lifetimes, resistance to photobleaching, ligand tunability, and efficient cellular uptake. Polypyridyl ruthenium(II) complexes have been extensively studied over the past few decades. Their octahedral coordination structure allows for excellent control of the spatial volume surrounding the ligand, making them more suitable for intermolecular interactions, such as electrostatic interactions between anions and cations, and interactions with solvent molecules. This has the potential to significantly enhance the sensitivity of phosphorescence to the surrounding environment. Furthermore, this structure imparts a high degree of symmetry and stability to ruthenium(II) complexes, allowing the design of compounds with different recognition functions by modifying the functional groups on the ligands.

[0004] Carbonate is commonly found in water and soil, affecting water quality and soil pH. Bicarbonate is commonly used in the human body for acid-base regulation, respiratory regulation, blood buffering, and calcium ion stabilization. Bicarbonate is the primary component that influences the alkalinity of drinking water. Therefore, changes in carbonate and bicarbonate concentrations are of great significance for environmental monitoring and biological acid-base balance. Bisulfite acts as a catalyst and stabilizer in acid-base reactions and has broad application prospects in food processing, the textile industry, and environmental protection. Hypochlorite is a commonly used bleaching agent, oxidant, and disinfectant. In the past, developing fluorescent probes to detect the presence and concentration changes of anions has also faced significant challenges. Traditional methods for detecting anions include titration, electrode methods, and inductively coupled plasma atomic spectrometry. However, these methods are often time-consuming or require expensive instrumentation, multiple reagents, and complex procedures. In contrast, colorimetric probe technology offers inherent advantages due to its simplicity, visualization, rapid response, real-time monitoring, and high sensitivity and selectivity. In the past decade, many fluorescent probes for detecting anions have been reported, but few colorimetric probes for anions have been reported. Therefore, the development of probes with colorimetric detection performance is of great significance. Summary of the Invention

[0005] In view of the shortcomings of the prior art, the present invention provides a type of polypyridine ruthenium complex, the cation of which is in, Selected from N is selected from N≡C-CH3; Selected from

[0006] The preparation method of the above-mentioned polypyridine ruthenium complex is as follows:

[0007] (1) Under an inert atmosphere, ruthenium trichloride is dissolved in N,N-dimethylformamide, and then a bipyridine compound and a lithium salt are added and mixed to obtain a mixed solution. The mixed solution is refluxed at 110-130°C for 8 hours. After the reaction product is cooled to room temperature, ice acetone is added and heated at 4°C overnight. The solid-liquid separation is performed, and the solid is washed and dried to obtain a ruthenium precursor I or a ruthenium precursor II.

[0008] Ruthenium Precursor I 2H2O or ruthenium precursor II 2H2O; among which bipyridine compounds are The lithium salt is lithium chloride; the molar ratio of ruthenium trichloride to the bipyridine compound is 1:1-3, and the molar ratio of ruthenium trichloride to the lithium salt is 1:7-8;

[0009] (2) Under an inert atmosphere, dissolving the ruthenium precursor I or ruthenium precursor II of step (1) in a mixed solution of ethanol and ultrapure water, adding an imidazole compound, and mixing to obtain a mixed solution. After the mixed solution is refluxed at 70-90° C. for 8 h, an anion exchanger is added and stirred at room temperature for 2 h. A solvent is added to precipitate, the solid-liquid separation is performed, and the solid is washed and dried to obtain a polypyridine ruthenium complex;

[0010] The imidazole compound is The molar ratio of ruthenium precursor I or ruthenium precursor II: imidazole compound is 1:1-2; the molar ratio of ruthenium precursor I or ruthenium precursor II: anion exchanger is 1:9-11;

[0011] Alternatively, under an inert atmosphere, the ruthenium precursor I or ruthenium precursor II in step (1) is placed in acetonitrile, a dechlorinating agent is added, and the mixture is stirred at room temperature for dechlorination for 24 hours. The solid-liquid separation is performed, and the filtrate is dried to obtain a polypyridine ruthenium complex (N is N≡C-CH3), wherein the molar ratio of ruthenium precursor I or ruthenium precursor II: dechlorinating agent is 1:1-3; and the dechlorinating agent is silver trifluoromethanesulfonate.

[0012] The inert atmosphere is N2, Ar or He.

[0013] Another object of the present invention is to apply the above-mentioned polypyridine ruthenium complex to the detection of carbonate, bicarbonate, bisulfite, and hypochlorite. Such polypyridine ruthenium complex is used as a colorimetric probe. When induced by carbonate, bicarbonate, bisulfite, and hypochlorite in an aqueous solution, the structure between the complex body and the ligand changes, causing the color of the complex solution to change, thereby achieving specific colorimetric detection of carbonate, bicarbonate, bisulfite, and hypochlorite. Test paper method, titration ultraviolet detection, and other methods can also be used.

[0014] Another object of the present invention is to use the above-mentioned polypyridine ruthenium complex in the preparation of cell diagnosis and treatment imaging reagents.

[0015] Another object of the present invention is to use the polypyridine ruthenium complex in the preparation of anti-tumor drugs.

[0016] The advantages and technical effects of the present invention are as follows:

[0017] The polypyridine ruthenium complex of the present invention not only has anti-tumor activity and can be used in the preparation of anti-tumor drugs, but also the polypyridine ruthenium (II) complex synthesized by the present invention undergoes structural changes between the complex body and the ligand under the induction of carbonate and bicarbonate, bisulfite and hypochlorite in the solution environment, and the color change of the solution of such substance can be clearly observed under natural light; and the polypyridine ruthenium (II) complex has the characteristics of high sensitivity, low detection cost, convenient operation, rapid determination and real-time detection. Therefore, such polypyridine ruthenium (II) complex can be applied to the fields of colorimetric detection of ions, tumor treatment and the like. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is the high-resolution mass spectrum of the complex Ru1;

[0019] Figure 2 The H NMR spectrum of the complex Ru2 is ( 1 H-NMR, d6-DMSO) pattern;

[0020] Figure 3 This is the high-resolution mass spectrum of the complex Ru2;

[0021] Figure 4 This is the high-resolution mass spectrum of the complex Ru3;

[0022] Figure 5 The H NMR spectrum of the complex Ru4 is ( 1 H-NMR, d6-DMSO) pattern;

[0023] Figure 6 This is the high-resolution mass spectrum of the complex Ru4;

[0024] Figure 7 This is the high-resolution mass spectrum of the complex Ru5;

[0025] Figure 8 The H NMR spectrum of the complex Ru6 is ( 1 H-NMR, d6-DMSO) pattern;

[0026] Figure 9 This is the high-resolution mass spectrum of the complex Ru6;

[0027] Figure 10 The fluorescence spectra of the complex Ru1 (40 μmol / L) (left) and the complex Ru2 (40 μmol / L) (right) after irradiation with 463 nm wavelength for different time periods;

[0028] Figure 11 The fluorescence spectra of the complex Ru3 (40 μmol / L) (left) and the complex Ru5 (40 μmol / L) (right) after irradiation with 463 nm wavelength for different time periods;

[0029] Figure 12 The UV spectra and color development of the complex Ru1 (40 μmol / L) (left) and the complex Ru2 (40 μmol / L) (right) after 10 min of illumination with the addition of different anions;

[0030] Figure 13 The UV spectra and color development of the complex Ru3 (40 μmol / L) (left) and the complex Ru5 (40 μmol / L) (right) after adding different anions.

[0031] Figure 14 The UV spectra and color development of the complex Ru3 (40 μmol / L) (left) and the complex Ru5 (40 μmol / L) (right) after 60 min of illumination with the addition of different anions;

[0032] Figure 15 The UV spectra and color development of the complex Ru4 (40 μmol / L) (left) and the complex Ru6 (40 μmol / L) (right) after adding different anions.

[0033] Figure 16 The fluorescence spectra (left) and histogram (right) of the complex Ru3 (40 μmol / L) with different anions added are shown.

[0034] Figure 17 The fluorescence spectra (left) and histogram (right) of the complex Ru5 (40 μmol / L) with different anions added are shown;

[0035] Figure 18The UV spectra and color development of the complex Ru2 (40 μmol / L) (left) and the complex Ru4 (40 μmol / L) (right) after 10 min of illumination with the addition of different carbonates.

[0036] Figure 19 The UV spectra and color development of the complex Ru5 (40 μmol / L) (left) and the complex Ru5 (40 μmol / L) after 60 min of illumination (right) when different carbonates were added;

[0037] Figure 20 The fluorescence spectra (left) and histogram (right) of the complex Ru5 (40 μmol / L) added with different carbonates are shown;

[0038] Figure 21 It is CO3 2- UV spectra of the titrated complexes Ru2 (60 μmol / L) (left) and Ru5 (60 μmol / L) (right);

[0039] Figure 22 The complex Ru5 (800μmol / L) is exposed to different concentrations of CO3 2- Filter paper detection diagram in the presence of;

[0040] Figure 23 The UV spectra of the complexes Ru3 (10 μmol / L) (left) and Ru5 (10 μmol / L) (right) in different solvents are shown;

[0041] Figure 24 The fluorescence spectra of the complexes Ru1 (10 μmol / L) (left) and Ru2 (10 μmol / L) (right) in different solvents are shown;

[0042] Figure 25 The fluorescence spectra of the complexes Ru3 (10 μmol / L) (left) and Ru5 (10 μmol / L) (right) in different solvents are shown;

[0043] Figure 26 The fluorescence spectra of the complexes Ru1 (50 μmol / L) (left) and Ru2 (30 μmol / L) (right) in mixed solutions of glycerol and water at different ratios are shown;

[0044] Figure 27 The fluorescence spectra of the complexes Ru3 (40 μmol / L) (left) and Ru5 (40 μmol / L) (right) in mixed solutions of glycerol and water at different ratios are shown;

[0045] Figure 28 It is the imaging of the complex Ru2 in tumor cells. Specific implementation methods

[0046] The present invention is further described in detail below through the examples, but the scope of protection of the present invention is not limited to the contents described herein. The structures of the compounds prepared in the following examples were confirmed by proton nuclear magnetic resonance spectroscopy and mass spectrometry. Where specific techniques or conditions are not specified in the examples, the methods described in the literature in the art or in the product instructions were used. Materials or equipment used without manufacturer indication are commercially available, and the methods used are conventional methods unless otherwise specified.

[0047] Example 1: Synthesis of polypyridine ruthenium complex Ru1

[0048] (1) Under nitrogen atmosphere, ruthenium trichloride is dissolved in N,N-dimethylformamide, and then 2,2'-bipyridine (bpy) and lithium chloride are added, wherein the molar ratio of ruthenium trichloride to 2,2'-bipyridine is 1:2, and the molar ratio of ruthenium trichloride to lithium chloride is 1:7.5. After mixing, the mixture is refluxed at 120°C for 8 hours, cooled to room temperature, added with a large amount of ice acetone, and refrigerated at 4°C overnight. In this process, a black precipitate will precipitate. After solid-liquid separation, the solid is washed with ice acetone and water in turn, and dried at 50°C to obtain a black solid, namely (bpy). 2- Ru precursor;

[0049] (2) Under nitrogen atmosphere, step (1) (bpy) 2- The Ru precursor was dissolved in a mixed solution of anhydrous ethanol and ultrapure water (10 mL of anhydrous ethanol and 10 mL of ultrapure water) at a molar ratio of 1:2. Imidazole was added and the mixture was refluxed at 80°C for 8 h. After cooling to room temperature, the mixture was heated to 100 °C and heated to 100 °C. 2- The Ru precursor: anion exchanger was added with ammonium hexafluorophosphate in a molar ratio of 1:10, stirred at room temperature for 2 hours, and then a large amount of ultrapure water was added and placed in a refrigerator at 4°C for 2 hours to precipitate. The solid was separated into liquid and washed with water and ethanol, and then dried at 50°C to obtain a red solid complex Ru1; the yield was 69.8%; its mass spectrum is shown in FIG. Figure 1 ;

[0050] The structural formula of the complex Ru1 is

[0051] NMR 1 The H spectrum and mass spectrum data are as follows: 1H NMR (600MHz, DMSO-d6) δ12.80(s,2H),8.95(d,J=5.6Hz,2H),8.64(d,J=8.2Hz,2H),8.56(d,J=8.1Hz,2H),8.1 5(t,J=7.9Hz,2H),7.96-7.88(m,4H),7.77(d,J=9.0Hz,4H),7.39(t,J=6.7Hz,2H),7.25(s,2H),6.70(s,2H).

[0052] Example 2: Synthesis of polypyridine ruthenium complex Ru2

[0053] (1) Same as step (1) in Example 1;

[0054] (2) Under nitrogen atmosphere, step (1) (bpy) 2- The Ru precursor was dissolved in a mixed solution of anhydrous ethanol and ultrapure water (10 mL of anhydrous ethanol and 10 mL of ultrapure water) at a molar ratio of 1:2. Ethylimidazole was added and the mixture was refluxed at 80°C for 8 h. After cooling to room temperature, the mixture was heated to 100 °C and heated to 100 °C. 2- Add ammonium hexafluorophosphate in a molar ratio of Ru precursor to anion exchanger of 1:10, stir at room temperature for 2 hours, add a large amount of ultrapure water and place in a refrigerator at 4°C for 2 hours to precipitate, separate the solid and liquid, wash the solid with water and ethanol, and dry it at 50°C to obtain a red solid complex Ru2; the yield is 67.9%; its H NMR spectrum is shown in Figure 2 , mass spectrum see Figure 3 ;

[0055] The structural formula of the complex Ru2 is

[0056] NMR 1 The H spectrum and mass spectrum data are as follows: 1 H NMR (600MHz, DMSO-d6) δ8.93(d,J=5.5Hz,2H),8.65(d,J=8.1Hz,2H),8.56(d,J=8.1Hz,2H),8.17(t,J=7.9Hz,2H),7.94(t,J=7. 8Hz,2H),7.86(d,J=5.7Hz,2H),7.84-7.75(m,4H),7.41-7.29(m,4H),6.68(s,2H),3.92(q,J=7.2Hz,4H),1.22(t,J=7.3Hz,6H).

[0057] Example 3: Synthesis of polypyridine ruthenium complex Ru3

[0058] (1) Same as step (1) in Example 1;

[0059] (2) Under nitrogen atmosphere, step (1) (bpy) 2- The Ru precursor was dissolved in a mixed solution of anhydrous ethanol and ultrapure water (10 mL of anhydrous ethanol and 10 mL of ultrapure water) at a molar ratio of 1:1. Biimidazole was added and the mixture was refluxed at 80°C for 8 h. After cooling to room temperature, the mixture was heated to 100 °C. 2- The Ru precursor: anion exchanger was added with ammonium hexafluorophosphate in a molar ratio of 1:10, stirred at room temperature for 2 hours, the reaction product was dried at 60°C, a small amount of acetonitrile was added to completely dissolve it, and then ether was added and placed in a refrigerator at 4°C for 2 hours to precipitate. The solid was separated into liquid, the solid was washed with ether, and dried at 50°C to obtain a red solid complex Ru3; the yield was 79%; its mass spectrum is shown in FIG. Figure 4 ;

[0060] The structural formula of the complex Ru3 is

[0061] NMR 1 The H spectrum and mass spectrum data are as follows: 1 H NMR (600MHz, DMSO-d6) δ8.77(t,J=7.2Hz,4H),8.16-8.04(m,4H),7.85(dd,J=30.2,5.6Hz,4H),7.63-7.55(m,2H),7.47(d,J=5.8Hz,4H),6.45(s,2H).

[0062] Example 4: Synthesis of polypyridine ruthenium complex Ru4

[0063] (1) Same as step (1) in Example 1;

[0064] (2) Under nitrogen atmosphere, step (1) (bpy) 2- The Ru precursor was dissolved in 15 mL of acetonitrile, and silver trifluoromethanesulfonate was added in a molar ratio of 1:2. The reaction was carried out at room temperature for 24 h, filtered, and the filtrate was rotary evaporated at 50 ° C to obtain a yellow concentrated liquid, which was dried at 50 ° C to obtain a yellow solid Ru4; the yield was 76.4%; its H NMR spectrum is shown in Figure 5 , mass spectrum see Figure 6 ;

[0065] The structural formula of the complex Ru4 is

[0066] NMR 1 The H spectrum and mass spectrum data are as follows: 1H NMR (600MHz, DMSO-d6) δ9.39(d,J=5.4Hz,2H),8.86(d,J=8.2Hz,2H),8.73(d,J=8.1Hz,2H),8.39(t,J=8.0Hz ,2H),8.07(t,J=7.9Hz,2H),7.96(t,J=6.6Hz,2H),7.60(d,J=5.6Hz,2H),7.40(t,J=6.7Hz,2H),2.48(s,6H).

[0067] Example 5: Synthesis of polypyridine ruthenium complex Ru5

[0068] (1) Under nitrogen atmosphere, ruthenium trichloride is dissolved in N,N-dimethylformamide, and then 1,10-phenanthroline (phen) is added, wherein the molar ratio of ruthenium trichloride to 1,10-phenanthroline is 1:2, and the molar ratio of ruthenium trichloride to lithium chloride is 1:7.5. The mixture is refluxed at 120°C for 8 hours. After the reaction product is cooled to room temperature, a large amount of ice acetone is added and the product is refrigerated at 4°C overnight. The solid is filtered, and the solid is washed with ice acetone and water in turn, and dried at 50°C to obtain a black solid, namely (phen). 2- Ru precursor;

[0069] (2) Under nitrogen atmosphere, the step (1) (phen) 2- The Ru precursor was placed in a mixed solution of anhydrous ethanol and ultrapure water (10 mL of anhydrous ethanol and 10 mL of ultrapure water), and biimidazole was added in a molar ratio of 1:1 and mixed. The mixture was refluxed at 80°C for 8 hours. After cooling to room temperature, ammonium hexafluorophosphate was added in a molar ratio of 1:10. After stirring at room temperature for 2 hours, the reaction product was spin-dried at 60°C, a small amount of acetonitrile was added to completely dissolve it, and a large amount of ether was added and placed in a refrigerator at 4°C for 2 hours to precipitate. The precipitate was filtered, and the solid was washed with ether and dried at 50°C to obtain a red solid complex Ru5; the yield was 80.6%; its mass spectrum is shown in FIG. Figure 7 ;

[0070] The structural formula of the complex Ru5 is

[0071] NMR 1 The H spectrum and mass spectrum data are as follows: 1H NMR (600MHz, DMSO-d6) δ8.79(d,J=8.2Hz,2H),8.65(d,J=8.1Hz,2H),8.40-8.34(m,4H),8.33(d,J=8.8Hz,2H ),8.10(d,J=5.3Hz,2H),8.00(dd,J=8.3,5.2Hz,2H),7.67(dd,J=8.2,5.3Hz,2H),7.47(s,2H),6.39(s,2H).

[0072] Example 6: Synthesis of polypyridine ruthenium complex Ru6

[0073] (1) Same as step (1) of Example 5;

[0074] (2) Under nitrogen atmosphere, the step (1) (phen) 2- The Ru precursor was dissolved in 15 mL of acetonitrile, and silver trifluoromethanesulfonate was added in a molar ratio of 1:2. The reaction was carried out at room temperature for 24 h, filtered, and the filtrate was rotary evaporated at 50 ° C to obtain a yellow concentrated liquid, which was dried at 50 ° C to obtain a yellow solid Ru6; the yield was 74.7%; its H NMR spectrum is shown in Figure 8 , mass spectrum see Figure 9 ;

[0075] The structural formula of the complex Ru6 is

[0076] NMR 1 The H spectrum and mass spectrum data are as follows: 1 H NMR(600MHz,DMSO-d6)δ9.86(d,J=5.0Hz,2H),9.06(d,J=8.1Hz,2H),8.64(d,J=8.0Hz,2H),8.46(dd,J=8.9,4.2Hz,2H), 8.36(dd,J=8.4,5.1Hz,2H),8.31(dd,J=9.0,3.8Hz,2H),7.85(d,J=5.1Hz,2H),7.61-7.56(m,2H),2.45(d,J=3.9Hz,6H).

[0077] Example 7: Ion Selectivity Test of Polypyridine Ruthenium (II) Complexes

[0078] 1. Fluorescence emission of polypyridine ruthenium (II) complexes under different illumination times

[0079] 1 mg of Ru1, Ru2, Ru3, Ru4, Ru5, and Ru6 complexes were weighed respectively, and dimethyl sulfoxide was added to prepare a 40 mmol / L mother solution. 3 μL of each was added to 6 5 mL centrifuge tubes, 27 μL of dimethyl sulfoxide was added, and the mixture was diluted with water to 40 μmol / L. Ru1 and Ru2 were irradiated with light of 463 nm for 0 s, 10 s, 30 s, 1 min, 5 min, and 10 min, and Ru3 and Ru5 were irradiated for 0 min, 1 min, 5 min, 10 min, 30 min, and 60 min, and fluorescence detection was performed respectively to obtain fluorescence emission spectra. The results are shown in the table. Figure 10 、 Figure 11 ;

[0080] From the figure, we can see that the fluorescence of Ru1 decreased by about 1.4 times after 1 minute of illumination at 463 nm wavelength and was completely quenched after 5 minutes of illumination; the fluorescence of Ru2 decreased by about 5.5 times after 1 minute of illumination and was completely quenched after 5 minutes of illumination; the fluorescence of Ru3 decreased by about 9 times after 60 minutes of illumination; the fluorescence of Ru5 decreased by about 30 times after 60 minutes of illumination;

[0081] The Ru4 and Ru6 complexes showed no fluorescence.

[0082] 2. Anion selectivity test of complexes Ru1, Ru2, Ru3, Ru4, Ru5, and Ru6

[0083] Weigh 1 mg of each Ru1, Ru2, Ru3, and Ru5 complex, add dimethyl sulfoxide to prepare a 40 mmol / L stock solution, take 3 μL of each and add it to 30 5 mL centrifuge tubes, add 27 μL of dimethyl sulfoxide, and dilute with water to 40 μmol / L. Fifteen of the centrifuge tubes were not illuminated, and 15 were illuminated at a wavelength of 463 nm until fluorescence quenching (10 min for Ru1 and Ru2 complexes, 60 min for Ru3 and Ru5 complexes).

[0084] Ru4 and Ru6 complexes were prepared into 40 μmol / L solutions in 15 centrifuge tubes according to the above method and were not exposed to light;

[0085] Take appropriate amount of H2PO4 - Br - 、SO3 2- 、F - 、CO3 2- 、ClO4 - , I - 、CH3COO - 、HSO3 - 、Cl - 、HCO3 -、SO4 2- , ClO - 、NO3 - The same anion was added to water to prepare a 5 mol / L stock solution, and 3 μL was taken and added to the centrifuge tubes of the above different compounds, and the final ion concentration was 5 mmol / L. At the same time, the solution without adding anions was used as a control (free). The above solution was detected by ultraviolet absorption in the range of 200-800 nm to obtain an ultraviolet spectrum, and the fluorescence spectrum was obtained by fluorescence detection in the range of 430-860 nm. The results are shown in Figure 12-17 ;

[0086] Ru1 after illumination in CO3 2- (yellow turns pink), HSO3 - (yellow turns light green), ClO - (yellow to pink) showed obvious color change in the presence of light; compared with the control, there was no color change in the absence of light;

[0087] Ru2 after illumination in CO3 2- (yellow turns pink), HSO3 - (yellow to green), ClO - (yellow to pink) showed obvious color change in the presence of light, while no color change was observed in the absence of light compared with the control;

[0088] Ru3 in SO3 2- (yellow to red), CO3 2- (yellow turns pink), HCO3 - (yellowish to reddish), ClO - The color changes significantly in the presence of SO3 2- (yellowish red (light)), CO3 2- (yellow turns pink), HCO3 - (yellowish red (light)), ClO - (yellow to reddish (light)) with obvious color reaction;

[0089] Ru4 in CO3 2- (yellow turns darker and reddish), HSO3 - (yellow turns lighter green), ClO - (yellow to reddish) with obvious color change in the presence of

[0090] Ru5 in SO3 2- (yellow turns darker), CO3 2- (yellow to red), HCO3 - (yellow turns darker), ClO -The color changes significantly in the presence of SO3 2- (yellow becomes darker (less obvious)), CO3 2- (yellow turns darker), HCO3 - (yellow turns darker), ClO - (yellow becomes darker) in the presence of an obvious color change;

[0091] Ru6 in CO3 2- (yellow turns darker and reddish), HSO3 - (color becomes lighter green), ClO - (yellowish to reddish) shows obvious color change.

[0092] 3. Experiment on the influence of cations on the detection of carbonate ions by complexes

[0093] 1 mg of Ru2, Ru4, and Ru5 complexes were weighed respectively, and dimethyl sulfoxide was added to prepare a 40 mmol / L mother solution. 3 μL of each solution was added to 14 5 mL centrifuge tubes, 27 μL of dimethyl sulfoxide was added, and the solution was diluted with water to 40 μmol / L. 7 tubes of Ru2 and Ru5 complex solutions were set up without light treatment, and 7 tubes were set up with 463 nm wavelength light treatment (Ru2 was illuminated for 10 minutes and Ru5 was illuminated for 60 minutes); the Ru4 complex solution was prepared to a concentration of 40 μmol / L, and carbonate was directly added to 7 tubes;

[0094] Take appropriate amounts of Na2CO3, CaCO3, ZnCO3, MgCO3, (NH4)2CO3, and K2CO3 and add water to prepare a 5 mol / L stock solution. Take 3 μL of each and add it to the centrifuge tubes of the above different compounds. The final ion concentration is 5 mmol / L. At the same time, a control without adding cations is used (free). The above solutions are detected by ultraviolet absorption in the range of 200-800 nm to obtain ultraviolet spectra, and by fluorescence detection in the range of 430-860 nm to obtain fluorescence spectra. The results are shown in Figure 2. Figure 18-20 ;

[0095] Compared with the control, Ru2 + (yellow to red), Mg 2+ (yellow becomes darker), K + (yellow to red) shows obvious color change in the presence of Na + Mg 2+ , K + The presence of light has no effect on the detection of carbonate ions, and color changes still occur; the solution of the complex Ru2 without light treatment has no color change or fluorescence change in the presence of different carbonates; compared with the control, the color change of Ru4 in the presence of carbonates is weaker ( Figure 18 );

[0096] Ru5 in Na + (yellow turns darker and reddish), Mg 2+ (yellow turns darker) and a significant color change occurs; Ru5 fluorescence is quenched to varying degrees in the presence of different carbonates, among which Na + , Ca 2+ Mg 2+ , K + In the presence of different carbonates, the fluorescence is completely quenched; the color change of Ru5 solution treated with light is weak in the presence of different carbonates ( Figure 19-20 );

[0097] The above shows that the presence of different cations has an impact on the response of carbonate ions.

[0098] 4. CO3 2- UV titration

[0099] Weigh 1 mg of Ru2 and Ru5 complexes respectively, add dimethyl sulfoxide to prepare a 60 mmol / L mother solution, take 3 μL of each into a 5 mL centrifuge tube, add 27 μL of dimethyl sulfoxide, dilute with water to 60 μmol / L, and irradiate the Ru2 solution with 463 nm wavelength light; take an appropriate amount of Na2CO3 and dissolve it in water to make a 10 mmol / L mother solution;

[0100] The Ru2 complex solution after illumination was subjected to UV detection to obtain a UV absorption spectrum of a Na2CO3 concentration of 0 μmol / L. 12 μL of Na2CO3 solution was added to the solution to obtain a solution with a Na2CO3 concentration of 40 μmol / L. The UV absorption spectrum at this Na2CO3 concentration was obtained by UV detection. Similarly, the UV absorption spectra of Ru2 complex solutions containing 0-2000 μmol / L Na2CO3 were detected. Similarly, the Na2CO3 solution was added dropwise to the Ru5 complex solution, and the UV absorption spectra of solutions with a Na2CO3 concentration of 0-160 μmol / L were detected. The results are shown in FIG. Figure 21 ;

[0101] In the solution of Ru2 complex after illumination, CO3 2- As the concentration increases, the ultraviolet absorption at 488nm decreases by about 1 times. When the intensity is 0.1au, the corresponding curve red-shifts from 529nm to 544nm, a total red-shift of 15nm.

[0102] In Ru5 solution (without light), with CO3 2- As the concentration increases, the ultraviolet absorption at 451nm decreases by about 1 times. When the intensity is 0.1au, the corresponding curve red-shifts from 495nm to 520nm, a total red-shift of 25nm.

[0103] 5. Complex Ru5 filter paper diagnosis

[0104] Weigh 0.5 mg of Ru5 complex and add water to prepare a solution with a concentration of 800 μmol / L. Take 1 mg of sodium carbonate to prepare solutions with concentrations of 10 mmol / L, 5 mmol / L, and 2.5 mmol / L. Cut the filter paper into 1 cm long and 0.5 cm wide pieces, take 10 μL of the complex solution and drop it in the middle of the filter paper. After the filter paper is dried, add CO3 at each concentration. 2- 10 μL of the solution was placed on filter paper, dried, and the color and fluorescence changes of the filter paper were observed under white light and fluorescence. Figure 22 ;

[0105] Complex Ru5 with CO3 2- As the concentration increases, the filter paper changes from yellow to pink under natural light, and the fluorescence of the complex is quenched under fluorescence.

[0106] Example 8: Solvent Effect

[0107] 1. Solvent effect of complexes Ru1, Ru2, Ru3, and Ru5

[0108] 1 mg of Ru1, Ru2, Ru3, and Ru5 complexes were weighed respectively, and dimethyl sulfoxide was added to prepare a 10 mmol / L mother solution. 3 μL of each solution was added to a 5 mL centrifuge tube, 27 μL of dimethyl sulfoxide was added, and then 3 mL of water, dimethyl sulfoxide, ethanol, acetonitrile, ethyl acetate, tetrahydrofuran, N,N-dimethylformamide, toluene, acetone, cyclohexane, dichloromethane, methanol, ether, and chloroform were added to the centrifuge tube to prepare a solution with a complex concentration of 10 μmol / L; the above solution was subjected to ultraviolet absorption detection in the range of 200-800 nm to obtain an ultraviolet spectrum; and fluorescence detection in the range of 430-860 nm to obtain a fluorescence emission spectrum; the results are shown in FIG. Figure 23-25 ;

[0109] Ru1 has strong fluorescence in dichloromethane and chloroform solutions. Compared with the complex in aqueous solution, the fluorescence in dichloromethane is enhanced by about 7 times, and the fluorescence in chloroform is enhanced by about 3 times;

[0110] Ru2 has strong fluorescence in dichloromethane, chloroform, and ethanol. Compared with the complex in aqueous solution, the fluorescence in dichloromethane is enhanced by about 7 times, the fluorescence in chloroform is enhanced by about 4 times, and the fluorescence in ethanol is enhanced by about 3 times.

[0111] Ru3 is red in dimethyl sulfoxide, N,N-dimethylformamide, and methanol. Compared with water, the red shift in dimethyl sulfoxide is from 479nm to 523nm, a total red shift of 44nm; in N,N-dimethylformamide, it is red shifted from 479nm to 522nm, a total red shift of 43nm; in methanol, it is red shifted from 479nm to 514nm, a total red shift of 35nm; Ru3 does not emit light in dimethyl sulfoxide, N,N-dimethylformamide, and methanol; its fluorescence is enhanced by about 4 times in dichloromethane and about 2 times in chloroform.

[0112] Ru5 is red in dimethyl sulfoxide, N,N-dimethylformamide, and methanol. Compared with water, the complex red-shifts from 473nm to 518nm in dimethyl sulfoxide, a total red-shift of 45nm; in N,N-dimethylformamide, it red-shifts from 473nm to 516nm, a total red-shift of 43nm; in methanol, it red-shifts from 473nm to 504nm, a total red-shift of 31nm; Ru5 does not emit light in dimethyl sulfoxide, N,N-dimethylformamide, and methanol; the fluorescence is enhanced by about 4 times in dichloromethane and about 2 times in chloroform.

[0113] 2. Viscosity effect

[0114] 1 mg of Ru1, Ru2, Ru3, and Ru5 complexes were weighed respectively, and dimethyl sulfoxide was added to prepare a 40 mmol / L mother solution. 3 μL of each solution was added to a 5 mL centrifuge tube, 27 μL of dimethyl sulfoxide was added, and 3 mL of glycerol-water mixture was added according to the glycerol content of 0%, 20%, 40%, 60%, 80%, and 100%. The solution was then subjected to fluorescence detection in the range of 430-860 nm to obtain a fluorescence emission spectrum. The results are shown in FIG. Figure 26 、 Figure 27 ;

[0115] In Ru1 solution, as the glycerol content increases, the fluorescence at 488 nm increases by about 39 times; the fluorescence at 639 nm increases by about 3 times;

[0116] In Ru2 solution, as the glycerol content increases, the fluorescence at 488nm increases by about 100 times; the fluorescence at 639nm increases by about 5 times;

[0117] In Ru3 solution, as the glycerol content increases, the fluorescence at 503nm increases by about 27.5 times; the fluorescence at 635nm increases by about 3 times;

[0118] In Ru5 solution, as the glycerol content increases, the fluorescence at 504nm increases by about 28 times; the fluorescence at 615nm increases by about 4 times.

[0119] Example 9: Application of polypyridyl ruthenium (II) complexes in cell imaging

[0120] HeLa cells (human cervical cancer cell line) in good growth condition were digested with trypsin, inoculated into confocal culture dishes, and cultured in an incubator containing 5% CO2 at 37°C. When the density of HeLa cells reached 70%, the complex Ru2 was added. The concentration of Ru2 in the incubation of cells was 40μmol / L. The cells in the culture dish were cultured for 2h and 4h. After incubation for 10-15min, the culture medium was removed, the cells were washed twice with PBS, and immediately observed with a laser confocal microscope. The results are shown in Figure 28 After culturing for another 2 hours and using an excitation wavelength of 631 nm, Ru2 emitted red fluorescence in the cells, but the red fluorescence did not colocalize well. After culturing for another 4 hours, the fluorescence was slightly enhanced, but there was still no colocalization. This experiment demonstrates that our ruthenium (II) complex can be used as a red fluorescence probe for imaging and tracing living cells in the future.

[0121] Example 10: Determination of antitumor activity of polypyridine ruthenium (II) complexes

[0122] The cytotoxicity of Ru1, Ru2, Ru3, Ru4, Ru5, and Ru6 complexes to HeLa (human cervical cancer cell line) was determined using the experimental group. The specific determination method is as follows:

[0123] The MTT colorimetric assay was used to determine the concentration of cells. The tumor cells were digested with trypsin to form single cell suspensions, and counted using a hemocytometer. The cell concentration was adjusted to 5 × 10 4 / mL, inoculated into 96-well plates, 160 μL per well, cultured for 24 h, then added with different concentrations of drugs, placed in a 5% CO2, 37°C incubator for 48 h and 72 h, and added MTT 20 μL / well 4 h before the end of incubation; after 4 h, discarded the supernatant, added DMSO 150 μL / well, and vibrated for 5 min. OD values ​​were measured using a microplate reader at a wavelength of 492 nm.

[0124] Calculate the survival rate of the tested tumor cells, plot and calculate IC 50 The antitumor activity of the complex was evaluated by PCR (Table 1). The complex had no antitumor activity after 48 h of co-incubation, but had antitumor activity after 72 h of co-incubation. Its toxicity was weaker than that of cisplatin.

[0125] Table 1 IC values ​​of polypyridine ruthenium (II) complexes prepared by the present invention 50 value

[0126]

Claims

1. Application of a polypyridine ruthenium complex in the preparation of anion detection reagents, characterized in that: For detecting CO3 2- 、HSO3 - , ClO - ; For detecting CO3 2- 、HSO3 - , ClO - ; For detection of SO3 2- 、CO3 2- 、HCO3 - , ClO - ; For detecting CO3 2- 、HSO3 - , ClO - ; For detection of SO3 2- 、CO3 2- 、HCO3 - , ClO - ; For detecting CO3 2- 、HSO3 - , ClO - .

2. Use of a polypyridine ruthenium complex in the preparation of a cell diagnosis and treatment imaging reagent, characterized in that: The polypyridine ruthenium complex is .

Images