A type of phosphorescent metal complex and its application

Abstract

The present invention discloses a class of phosphorescent metal complexes. Experiments have found that, in the presence of different anions, cations, and solvents, the fluorescence of these phosphorescent metal complexes has different degrees of quenching or enhancing effects, that is, the phosphorescent metal complexes can specifically recognize different anions and cations. Therefore, the phosphorescent metal complexes can be used as probes for fluorescence on / off quenching of specific ion species and solution fluorescence probes. They have application prospects in the fields of preparing test strips, fluorescent colorimetric anti-counterfeiting optical materials, and have the characteristics of high sensitivity, low detection cost, easy operation, rapid determination, and real-time detection. The phosphorescent metal complexes of the present invention have an aggregation-induced emission effect and can be used in the preparation of cell diagnosis and treatment imaging reagents. In anti-tumor activity detection, it was found that the phosphorescent metal complexes of the present invention have the potential to prepare anti-tumor drugs.

Description

Technical Field

[0001] The invention relates to a type of phosphorescent metal complex and its application in detecting ion solutions, biomedicine and bioimaging, belonging to the field of biotechnology. Background Art

[0002] Transition metal complexes are considered promising phosphorescent sensors for metal ions, anions, and biomolecules due to their attractive photophysical and chemical properties, including large Stokes shifts, long luminescence lifetimes, resistance to photobleaching, ligand tunability, and efficient cellular uptake. The photophysical properties of square-like planar palladium(II) / platinum(II) complexes have been extensively studied over the past few decades due to their intriguing spectral and luminescence properties, as well as their metal-metal interactions. Fluorescent soft salt complexes are optoelectronic materials composed of oppositely charged metal complexes bound by electrostatic and van der Waals forces, with their emission spectra typically consisting of two distinct components. Compared to single-component fluorescent metal complexes, fluorescent soft salt complexes contain a richer set of excited states, and energy transfer processes occur between the different components. In recent years, these complexes, as an emerging class of optoelectronic materials, have shown promising applications in various optoelectronic fields and have made considerable progress, such as in organic light-emitting diodes, bioimaging, photodynamic therapy, and electroluminescence color change.

[0003] Metal ions are an important component in maintaining the balanced permeability of multiple systems and are also necessary components of a wide range of enzyme reactions. The catalytic activity of some enzymes requires metal ions in addition to the protein part, that is, metal ions are components of the enzyme active center. Therefore, it is very necessary and valuable to design fluorescent probes to detect various metal elements. Summary of the Invention

[0004] The present invention provides a class of phosphorescent metal complexes, which are metal complexes based on platinum (II) and palladium (II) and phosphorescent soft salt compounds based on platinum-palladium, palladium-palladium and platinum-platinum complexes formed by the interaction of the metal complexes.

[0005] The phosphorescent metal complex is a cation a compound wherein Selected from Selected from

[0006] The phosphorescent metal complex is an anion of a compound wherein Selected from

[0007] The preparation method of the above-mentioned phosphorescent metal complex is as follows:

[0008] (1) Under an inert atmosphere, potassium tetrachloroplatinate and an aromatic ring compound are placed in a solvent and mixed, and stirred at 80° C. for 24-48 hours. The reaction product is concentrated, added with ultrapure water, mixed, and then ice-bathed until precipitation occurs. The solid is separated from the liquid to obtain a solid, and the solid is washed and dried to obtain a platinum-chloride bridged dimer;

[0009] Under an inert atmosphere, palladium acetate and an aromatic ring compound are dissolved in an alcohol solvent and mixed. The mixture is stirred at room temperature for 24-48 hours. The reaction product is spin-dried and recrystallized with dichloromethane and petroleum ether. An ice bath is performed and solid-liquid separation is performed to obtain a solid. The solid is washed and dried to obtain a palladium precursor. The palladium precursor and a lithium salt are placed in an acetone-water mixture (volume ratio of 13:4-7) and mixed. The mixture is stirred at room temperature for reaction. The solid-liquid separation is performed to collect a precipitate. The precipitate is dried to obtain a palladium-chloro bridged dimer, wherein the molar ratio of the palladium precursor to the lithium salt is 1:6-20.

[0010] The molar ratio of the potassium tetrachloroplatinate or palladium acetate to the aromatic ring compound is 1:1-3;

[0011] The aromatic ring compound is selected from

[0012] (2) Under an inert atmosphere, the platinum-chloride bridged dimer or the palladium-chloride bridged dimer and the ligand are placed in dichloromethane and mixed evenly, the mixture is reacted at room temperature to 50° C., and a metal platinum complex or a metal palladium complex is obtained by separation and purification;

[0013] The ligands are tetrabutylammonium cyanide and ethylenediamine, and the molar ratio of the platinum chloride bridge precursor or the palladium chloride bridge precursor to the ligand is 1:2-1:4.

[0014] The phosphorescent soft salt metal complex is selected from any one of the following structural formulas:

[0015]

[0016] in Selected from Selected from

[0017] The preparation method of the above-mentioned phosphorescent soft salt metal complex is as follows:

[0018] Under an inert atmosphere, a metal platinum complex and a metal palladium complex are added to an alcohol solvent, ultrasonically reacted at room temperature for 30 minutes, deionized water is added to the reaction product, ultrasonicated for another 30 minutes, solid-liquid separation is performed, and the solid is washed with deionized water and dichloromethane in sequence, and then recrystallized and purified with methanol / diethyl ether to obtain a yellow, yellow-green or red phosphorescent soft salt metal complex.

[0019] The inert atmosphere is N2, Ar or He.

[0020] Another object of the present invention is to apply the above phosphorescent metal complexes to anion detection.

[0021] The compound For quenching detection of NO2 - , ClO - , I - ; For enhanced detection of HSO3 - , ClO - ; For enhanced detection of CH3COO - , ClO - , I - 、NO3 - 、Cl - 、ClO4 - 、SO4 2- 、H2PO4 - ; For enhanced detection of ClO - 、HSO3 - ; For quenching detection of NO2 - , ClO - 、HSO3 - , I - ; For quenching detection of CO3 2- , ClO - ; For enhanced detection of ClO - 、HCO3 - ; For quenching detection of NO2 - , ClO - 、HSO3 - , I - .

[0022] Another object of the present invention is to apply the above phosphorescent metal complexes in cation detection.

[0023] The compound

[0024]

[0025] For quenching detection ;

[0026]

[0027] For enhanced detection ;

[0028]

[0029] For enhanced detection

[0030]

[0031] For enhanced detection

[0032] For quenching detection of Hg + 、Fe 3+ Enhanced detection of Sn 2+ ;

[0033]

[0034] For enhanced detection of Cu + 、Zn 2+ 、Ag + ;

[0035]

[0036] For enhanced detection of Sn 2+ 、Cu + 、Ag + 、Zn 2+ ;

[0037]

[0038] For quenching detection of Hg + 、Fe 3+ 、Cu + Enhanced detection of Sn 2+ 、Zn 2+ 、Ag + .

[0039] Another object of the present invention is to use the above-mentioned phosphorescent metal complex in the preparation of cell diagnosis and treatment imaging reagents, and the complex has an aggregation-induced emission effect.

[0040] Another object of the present invention is to use the above phosphorescent metal complex in the preparation of anti-tumor drugs.

[0041] Advantages and beneficial effects of the present invention:

[0042] 1. The phosphorescent metal complex of the present invention can induce electrostatic interaction with one or more of hypochlorite ions, zinc ions, monovalent copper ions, trivalent iron ions, mercury ions, silver ions, and tin ions in a solution environment at room temperature, causing a significant change in fluorescence. The complex can be used as a fluorescence on / off quenching probe for hypochlorite ions, zinc ions, monovalent copper ions, trivalent iron ions, mercury ions, silver ions, and tin ions. They have the characteristics of high sensitivity, low detection cost, easy operation, rapid determination, and real-time detection for the detection of ions in aqueous solution in vitro. In addition, test paper with the complex can be used for the detection of metal ions, and the naked eye can directly judge whether the ions exist or not. It has application prospects in the preparation of test paper, fluorescent color development anti-counterfeiting materials and other fields;

[0043] 2. The phosphorescent metal complexes of the present invention exhibit aggregation-induced emission effects and can aggregate and self-assemble in aqueous media to form nanoparticles. One type of phosphorescent metal complex exhibits a red-shifted absorption and emission spectrum, emitting yellow-red fluorescence at 500-700nm upon excitation by a 360-450nm light source; another type of phosphorescent metal complex emits green fluorescence at 500-700nm upon excitation by a 360-450nm light source. The nanoparticles self-assembled from these complexes can enter cells via endocytosis, enabling cell monitoring while exerting anti-tumor effects. They have strong resistance to photobleaching and good stability. Therefore, the design of these complexes provides application methods not only for in vitro and in vivo detection, but also for in vivo imaging studies.

[0044] 3. The four soft salts also show different fluorescence in different solvents and can be used to detect different solvents;

[0045] 4. Phosphorescent metal complexes have anti-tumor activity and can exist stably in the body environment, and can be used in the preparation of anti-tumor drugs; therefore, such phosphorescent complexes can be used in the fields of ion detection fluorescent probes, solvent detection probes, tumor treatment and fluorescent color development materials. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 This is the high-resolution mass spectrum of complex 1a;

[0047] Figure 2 This is the high-resolution mass spectrum of complex 1b;

[0048] Figure 3 This is the high-resolution mass spectrum of complex 2a;

[0049] Figure 4 This is the high-resolution mass spectrum of complex 2b;

[0050] Figure 5 is the H NMR spectrum of soft salt SA ( 1H-NMR, d6-DMSO) pattern;

[0051] Figure 6 is the nuclear magnetic resonance hydrogen spectrum of soft salt SB ( 1 H-NMR, d6-DMSO) pattern;

[0052] Figure 7 This is the result of the anion selectivity test of complexes 1a (20 μmol / L) and 1b (20 μmol / L);

[0053] Figure 8 This is the result of the anion selectivity test of complexes 2a (40 μmol / L) and 2b (40 μmol / L);

[0054] Figure 9 This is the result of the anion selectivity test of soft salts SA (20 μmol / L) and SB (40 μmol / L);

[0055] Figure 10 These are the results of the anion selectivity test using soft salts SC (20 μmol / L) and SD (20 μmol / L);

[0056] Figure 11 This is the result of the cation selectivity test of complexes 1a (20 μmol / L) and 1b (20 μmol / L);

[0057] Figure 12 This is the result of the cation selectivity test of complexes 2a (40 μmol / L) and 2b (40 μmol / L);

[0058] Figure 13 This is the result of the cation selectivity test of soft salts SA (20 μmol / L) and SB (40 μmol / L);

[0059] Figure 14 This is the result of the cation selectivity test of soft salt SC (20 μmol / L) and SD (20 μmol / L);

[0060] Figure 15 It is the Zn of complexes 2a, SB, and SD 2+ 、Cu + 、Ag + Ion test filter paper test results;

[0061] Figure 16 It is Zn 2+ Titrate complex 2a (40 μmol / L), soft salt SB (40 μmol / L) and Ag + Fluorescence spectrum of titration of soft salt SD (20 μmol / L);

[0062] Figure 17 is the AIE effect diagram of complexes 1a (20 μmol / L) and 2a (40 μmol / L);

[0063] Figure 18 is the AIE effect diagram of soft salt SA (20 μmol / L) and soft salt SB (40 μmol / L);

[0064] Figure 19 is the AIE effect diagram of soft salt SC (20 μmol / L) and soft salt SD (20 μmol / L);

[0065] Figure 20 is the fluorescence emission spectra of complexes 1a (20 μmol / L) and 1b (20 μmol / L) after reacting with GSH for 2 h;

[0066] Figure 21 is the fluorescence emission spectra of complexes 2a (40 μmol / L) and 2b (40 μmol / L) after reacting with GSH for 2 h;

[0067] Figure 22 This is the fluorescence emission spectrum of soft salt SA (20 μmol / L) and soft salt SB (40 μmol / L) after 2 hours of interaction with GSH;

[0068] Figure 23 This is the fluorescence emission spectrum of soft salt SC (20 μmol / L) and soft salt SD (20 μmol / L) after 2 hours of interaction with GSH;

[0069] Figure 24 This is the imaging result of phosphorescent soft salt complex SB in tumor cells;

[0070] Figure 25 is the fluorescence emission spectra of soft salt SA (20 μmol / L) and soft salt SB (40 μmol / L) in different solvents;

[0071] Figure 26 is the fluorescence emission spectra of soft salt SC (20 μmol / L) and soft salt SD (20 μmol / L) in different solvents;

[0072] Figure 27 is the fluorescence emission graph of soft salt SA (20 μmol / L) and soft salt SB (40 μmol / L) in response to amino acids;

[0073] Figure 28 is the fluorescence emission graph of soft salt SC (20 μmol / L) and SD (20 μmol / L) in response to amino acids;

[0074] Figure 29The fluorescence emission graphs of soft salt SA (20 μmol / L) and soft salt SB (40 μmol / L) in buffer solutions with different pH values ​​are shown in Figure 2.

[0075] Figure 30 These are the fluorescence emission graphs of soft salt SC (20 μmol / L) and SD (20 μmol / L) in buffer solutions of different pH values. DETAILED DESCRIPTION

[0076] The present invention is further described in detail below by way of examples, but the protection scope of the present invention is not limited to the contents described above.

[0077] The structures of the compounds prepared in the examples of the present invention were confirmed using proton nuclear magnetic resonance spectroscopy and mass spectrometry. Where specific techniques or experimental conditions are not specified in the examples, the procedures were performed according to those described in literature in the art or according to the product specifications. Materials or equipment used, where the manufacturer is not specified, are commercially available, and the methods used are conventional methods unless otherwise noted.

[0078] Example 1: Synthesis of complex 1a

[0079] (1) Under nitrogen atmosphere, potassium tetrachloroplatinate (100 mg, 0.24 mmol) and 2-phenylpyridine (74.8 mg, 0.48 mmol) were mixed and added to a mixture of ethylene glycol ether and water (14 mL of ethylene glycol ether and 7 mL of water) to dissolve the mixture, and the mixture was stirred at 80°C for 48 h. After the reaction, the reaction product was concentrated to 1-2 mL at 60°C, ultrapure water was added, and the mixture was ice-bathed until a large amount of precipitation occurred. Solid-liquid separation was performed to obtain a solid, which was washed with water and ether in sequence and dried in vacuo at 50°C to obtain a platinum-chloride bridged dimer [Pt(ppy)Cl]2 (134.9 g, 72.9%).

[0080] (2) Under nitrogen atmosphere, the platinum-chloride bridged dimer [Pt(ppy)Cl]2 (134.9 mg, 0.18 mmol) obtained in step (1) and ethylenediamine (43.3 mg, 0.72 mmol) were added to 15 mL of dichloromethane and mixed. The mixture was reacted and dechlorinated at 50°C with stirring for 5 h. After cooling, the mixture was extracted three times with dichloromethane and water. The dichloromethane phases were collected and combined, and finally purified by alumina column chromatography to obtain a yellow or green metal platinum complex 1a; the yield was 73.7%, and its high-resolution mass spectrum was shown in FIG. Figure 1 ;

[0081] The structural formula of complex 1a is

[0082] NMR 1 The H spectrum and mass spectrum data are as follows: 1H NMR(600MHz,DMSO-d6)δ8.62(d,J=5.8Hz,1H),8.14-8.03(m,2H),7.74-7.69(m,1H),7.37(t ,J=6.6Hz,1H),7.31-7.24(m,1H),7.15-7.09(m,2H),6.09(s,2H),5.32(s,2H),2.67(s,4H).

[0083] Example 2: Synthesis of Complex 1b

[0084] (1) Under nitrogen atmosphere, palladium acetate (80 mg, 0.36 mmol) and 2-(2,4-difluorophenyl)pyridine (68.1 mg, 0.36 mmol) were added to 15 mL of methanol and mixed. The mixture was stirred at room temperature for 24 h. During the reaction, a yellow solid precipitated. After the reaction, the reaction product was dried at 45 ° C to obtain a yellow solid. The yellow solid was dissolved in 2 mL of CH2Cl2, and then petroleum ether was added for precipitation. The mixture was placed in an ice bath for two hours, and the solid was collected by solid-liquid separation. The solid was washed with petroleum ether and ether in turn, and dried in vacuo at 50 ° C to obtain a bright yellow palladium acetate bridged dimer [Pd(2F-ppy)OAc]2 (190.4 mg, 75.3%).

[0085] (2) Under nitrogen atmosphere, [Pd(2F-ppy)OAc]2 (100 mg, 0.14 mmol) and lithium chloride (38.8 mg, 0.91 mmol) were added to a mixture of acetone and ultrapure water (acetone 26 mL, water 10 mL), mixed and reacted at room temperature for 12 h. The solid-liquid separation was carried out, and the solid was vacuum dried at 50°C to obtain a yellow palladium chloride bridged dimer [Pd(2F-ppy)Cl]2 (93.2 mg 78.6%).

[0086] (3) Under nitrogen atmosphere, palladium chloride bridged dimer [Pd(2F-ppy)Cl]2 (92.2 mg, 0.14 mmol) and tetrabutylammonium cyanide (113.5 mg, 0.42 mmol) were added to 30 mL of dichloromethane and mixed. The mixture was stirred and refluxed at 50°C for 5 h to obtain a yellow liquid crude product. The liquid was concentrated to 1-2 mL and then recrystallized by adding ether. Finally, the precipitate was washed with ether several times to obtain a yellow / off-white metal palladium complex 1b; the yield was 65.2%; its high-resolution mass spectrum is shown in FIG. Figure 2 ;

[0087] The structural formula of complex 1b is

[0088] NMR 1 The H spectrum and mass spectrum data are as follows: 1H NMR (600MHz, DMSO-d6) δ9.12(d,J=118.2Hz,1H),8.06(d,J=25.7Hz,2H),7.47(s,2H),6.96( s,1H),3.22-3.06(m,8H),1.63-1.51(m,8H),1.30(h,J=7.3Hz,8H),0.93(t,J=7.3Hz,12H).

[0089] Example 3: Synthesis of complex 2a

[0090] (1) Under nitrogen atmosphere, potassium tetrachloroplatinate (100 mg, 0.24 mmol) and 2-(2,4-difluorophenyl)pyridine (92.1 g, 0.48 mmol) were mixed and added to a mixture of ethylene glycol ether and water (ethylene glycol ether 14 mL, water 7 mL) and mixed. The mixture was stirred at room temperature for 48 h and then stirred at 80°C for 48 h. After the reaction, the reaction solution was concentrated to 1-2 mL at 60°C, and then ultrapure water was added and ice-bathed until a large amount of precipitation appeared. Solid-liquid separation was performed to obtain a solid, which was washed with water and ether in sequence and dried in vacuo at 50°C to obtain a platinum-chloride bridged dimer [Pt(2F-ppy)Cl]2 (154.8 mg, 76.5%).

[0091] (2) Under nitrogen atmosphere, the platinum-chloride bridged dimer [Pt(2F-ppy)Cl]2 (100 mg, 0.18 mmol) obtained in step (1) and tetrabutylammonium cyanide (193.3 mg, 0.72 mmol) were added to 15 mL of dichloro solvent and mixed evenly. The mixture was stirred at 50°C for dechlorination for 5 h. After cooling, the mixture was extracted three times with dichloromethane and water. The dichloromethane phases were collected and combined, and finally purified by alumina column chromatography to obtain a yellow or green metal platinum complex 2a with a yield of 75.3%. Its high-resolution mass spectrum is shown in FIG. Figure 3 ;

[0092] The structural formula of complex 2a is

[0093] NMR 1 The H spectrum and mass spectrum data are as follows: 1 H NMR(600MHz,DMSO-d6)δ9.29(d,J=5.4Hz,1H),8.18-8.06(m,2H),7.50(dt,J=12.9,12.4Hz,2H),6 .90(t,J=10.9Hz,1H),3.21-3.10(m,8H),1.55(s,8H),1.34-1.25(m,8H),0.92(t,J=7.2Hz,12H).

[0094] Example 4: Synthesis of complex 2b

[0095] (1) Under nitrogen atmosphere, palladium acetate (80 mg, 0.36 mmol) and 2-phenylpyridine (55.9 mg, 0.36 mmol) were added to 15 mL of methanol and mixed well. The mixture was stirred at room temperature for 24 h. During the reaction, a yellow solid precipitated. After the reaction, the reaction solution was dried at 45°C to obtain a yellow solid. The yellow solid was added to 1-2 mL of CH2Cl2, and then petroleum ether was added for precipitation. The mixture was ice-bathed for two hours, and the solid was collected by solid-liquid separation. The solid was washed with petroleum ether and ether in turn, and dried in vacuo at 50°C to obtain a bright yellow palladium acetate bridged dimer [Pd(ppy)OAc]2 (180.3 mg, 78.5%).

[0096] (2) Under nitrogen atmosphere, [Pd(ppy)OAc]2 (100 mg, 0.16 mmol) and lithium chloride (44.1 mg, 1.04 mmol) were added to a mixture of acetone and ultrapure water (acetone 26 mL, water 10 mL) and mixed. The mixture was reacted at room temperature for 12 h. The solid-liquid phase was separated and the solid was dried in vacuo at 50°C to obtain a yellow palladium chloride bridged dimer [Pd(ppy)Cl]2 (65 mg, 68.9%).

[0097] (3) Under nitrogen atmosphere, palladium chloride bridged dimer [Pd(ppy)Cl]2 (65 mg, 0.11 mmol) and tetrabutylammonium cyanide (259.5 mg, 0.44 mmol) were added to 30 mL of dichloromethane and mixed. The mixture was stirred and refluxed at 50°C for 5 h to obtain a yellow / off-white precipitate as a crude product. The precipitate was washed with dichloromethane several times to obtain a yellow / off-white metal palladium complex 2b; the yield was 74.9%; its high-resolution mass spectrum is shown in FIG. Figure 4 ;

[0098] The structural formula of the complex 2b is

[0099] NMR 1 The H spectrum and mass spectrum data are as follows: 1 H NMR(600MHz, Deuterium Oxide)δ8.40-8.27(m,2H),8.16-8.03(m,2H),7.85-7.73(m,2H),7.67(ddd,J=7.2,5.7,1.4Hz, 1H),7.58-7.49(m,1H),,3.40(dd,J=7.4,4.4Hz,2H),3.43-3.29(m,5H),2.50(t,J=1.7Hz,1H).

[0100] Example 5: Synthesis of soft salt SA

[0101] Under a nitrogen atmosphere, complex 1a and complex 1b were added to 5 mL of ethanol in a molar ratio of 1:1. The mixture was sonicated at room temperature for 30 minutes. 50 mL of deionized water was added to the yellow reaction product, and the mixture was sonicated for another 30 minutes. The solid-liquid separation was performed, and the precipitate was washed three times with a large amount of deionized water and dichloromethane, and then recrystallized and purified from methanol / diethyl ether (volume ratio 1:20) to obtain a yellow solid with a yield of 76.3%. The H NMR spectrum of the yellow solid is shown in FIG. Figure 5 ;

[0102] The structural formula of soft salt SA is

[0103] NMR 1 The H spectrum and mass spectrum data are as follows: 1 H NMR (600MHz, DMSO-d6) δ9.03(d,J=5.5Hz,1H),8.58(d,J=5.8Hz,1H),8.15-8.02(m,4H),7.74-7.68(m,1H),7.50-7.41( m,2H),7.37(t,J=6.6Hz,1H),7.28-7.23(m,1H),7.15-7.09(m,2H),6.90(s,1H),6.06(s,2H),5.25(s,2H),2.67(s,4H).

[0104] Example 6: Synthesis of soft salt SB

[0105] Under nitrogen atmosphere, complex 2a and complex 2b were added to 5 mL of ethanol in a molar ratio of 1:1 and mixed. The mixture was sonicated at room temperature for 30 minutes. 50 mL of deionized water was added to the yellow reaction product, and the mixture was sonicated for another 30 minutes. The solid-liquid separation was carried out, and the solid was washed three times with deionized water and dichloromethane, and then recrystallized and purified with methanol / diethyl ether (volume ratio 1:20) to obtain a yellow solid with a yield of 78.6%. Its H NMR spectrum is shown in FIG. Figure 6 ;

[0106] The structural formula of soft salt SB is

[0107] NMR 1 The H spectrum and mass spectrum data are as follows: 1H NMR(600MHz, DMSO-d6)δ9.29(d,J=5.7Hz,1H),8.32(d,J=5.5Hz,1H),8.19-8.05(m,4H),7.83-7.68(m,1H),7. 59-7.37(m,3H),7.19-6.86(m,4H),5.20(t,J=5.5Hz,2H),4.43(t,J=5.6Hz,2H),2.72(dq,J=28.6,5.7Hz,4H).

[0108] Example 7: Synthesis of Soft Salt SC

[0109] Under nitrogen atmosphere, complex 1b and complex 2b were added to 5 mL of ethanol in a molar ratio of 1:1 and mixed. The mixture was sonicated at room temperature for 30 minutes. 50 mL of deionized water was added to the yellow / off-white reaction product, and sonicated for another 30 minutes. The solid-liquid separation was performed, and the solid was washed three times with deionized water and dichloromethane, and then recrystallized from methanol / diethyl ether (volume ratio 1:20) to obtain a yellow-green product with a yield of 77.7%.

[0110] The structural formula of soft salt SC is

[0111] NMR 1 The H spectrum and mass spectrum data are as follows: 1 H NMR(600MHz,DMSO-d6)δ9.02(s,1H),8.35(dd,J=28.0,5.6Hz,1H),8.15(t,J=7.9Hz,1H),8 .08(tt,J=15.3,8.7Hz,4H),7.76-7.69(m,1H),7.52-7.46(m,1H),7.44-7.39(m,1H),7.17- 7.10(m,1H),7.06(dd,J=14.4,8.0Hz,2H),6.95-6.74(m,1H),5.31(d,J=6.4Hz,1H),5.20( d,J=6.7Hz,1H),4.60(t,J=5.6Hz,1H),4.44(d,J=6.2Hz,1H),2.72(dq,J=34.0,5.5Hz,4H).

[0112] Example 8: Synthesis of soft salt SD

[0113] Under nitrogen atmosphere, complex 1a and complex 2a were added to 5 mL of ethanol in a molar ratio of 1:1 and mixed. The mixture was sonicated at room temperature for 30 minutes. 50 mL of deionized water was added to the red reaction product, and sonication was continued for another 30 minutes. The solid-liquid separation was performed, and the solid was washed three times with deionized water and dichloromethane, followed by recrystallization and purification from methanol / diethyl ether (volume ratio 1:20) to obtain a red solid with a yield of 75.8%.

[0114] The structural formula of soft salt SD is

[0115] NMR 1 The H spectrum and mass spectrum data are as follows: 1 H NMR (600MHz, DMSO-d6) δ9.30(d,J=5.7Hz,1H),8.58(d,J=5.8Hz,1H),8.18-8.03(m,4H),7.71(dd,J=6.2,2.8Hz,1H),7.51(dd,J=8.5,2.5Hz,1H), 7.47(t,J=6.5Hz,1H),7.37(t,J=6.1Hz,1H),7.29-7.21(m,1H),7.16-7. 08(m,2H),6.89(d,J=3.2Hz,1H),6.06(s,2H),5.25(s,2H),2.67(s,4H).

[0116] Example 9: Selective Detection of Anions and Cations by Phosphorescent Metal Complexes

[0117] 1. Anion-selective detection of complexes 1a, 1b, 2a, 2b and phosphorescent soft salt complexes SA, SB, SC, SD

[0118] Weigh 1 mg each of complexes 1a, 1b, 2a, 2b and soft salts SA, SB, SC, and SD, add them to dimethyl sulfoxide to make a 20 mmol / L mother solution, take 3 μL of the mother solution into a 3 mL centrifuge tube, add 27 μL of dimethyl sulfoxide, and then dilute with water to a concentration of 20 μmol / L or 40 μmol / L, and add F - 、Cl - Br - , I - 、NO2 - 、NO3 - 、SO4 2- 、HSO3 - 、CO3 2- 、HCO3 - 、H2PO4 - 、CH3COO - , ClO - 、ClO4- The final concentration of ions was 5 mmol / L. The fluorescence of the above solution was measured under 320 nm / 400 nm excitation light to obtain the fluorescence spectrum. The results are shown in Figure 7-10 ;

[0119] It can be seen from the figure that complex 1a can specifically detect NO2 - , ClO - , I - , in NO2 - , ClO - , I - Green fluorescence was significantly quenched in the presence of HSO3; complex 1b can specifically detect HSO3 - , ClO - , in HSO3 - , ClO - The blue fluorescence of complex 2a was significantly enhanced in the presence of CH3COO - , ClO - , I - 、NO3 - 、Cl - 、ClO4 - 、SO4 2- 、H2PO4 - The orange fluorescence is enhanced in the presence of I - 、NO3 - 、SO4 2- In the presence of ClO, the emission is obviously red-shifted, and the fluorescence red-shift is 57nm; complex 2b is in the presence of ClO - 、HSO3 - In the presence of , the blue fluorescence is enhanced, and the intensity increases by about 2 times.

[0120] Soft salt SA and SD can specifically detect NO2 - , ClO - 、HSO3 - , I - , in NO2 - , ClO - 、HSO3 - , I - Green fluorescence is significantly quenched in the presence of SB; soft salt SB can specifically detect CO3 2- , ClO - , in CO3 2- , ClO - The red fluorescence is quenched in the presence of ClO; - 、HCO3 - The blue fluorescence is enhanced in the presence of

[0121] 2. Cation-selective detection of complexes 1a, 1b, 2a, 2b and phosphorescent soft salt complexes SA, SB, SC, SD

[0122] Weigh 1 mg each of complexes 1a, 1b, 2a, 2b and soft salts SA, SB, SC, and SD, add them to dimethyl sulfoxide to make a 20 mmol / L mother solution, take 3 μL of the mother solution into a 3 mL centrifuge tube, add 27 μL of dimethyl sulfoxide, and then use

[0123] 2+ water was diluted to 20μmol / L or 40μmol / L, and Hg + 、Cu + 、Fe 3+ NH4 + 、Co 2+ 、Fe 2+ 、Na + , Ca 2 + , Rb 2+ 、Cd 2+ 、Sn 2+ 、Cs + , K + 、Zn 2+ Mg 2+ 、Al 3+ 、Ba 2+ 、Li + 、Cu 2+ 、Ag + The final concentration of ions was 5 mmol / L. The fluorescence of the above solution was measured under 320 nm / 400 nm excitation light to obtain the fluorescence spectrum. The results are shown in Figure 11-14 ;

[0124] It can be seen from the figure that complex 1a in Hg + 、Cu + 、Fe 3+ Green fluorescence was significantly quenched in the presence of Zn 2+ Green fluorescence emission is enhanced in the presence of Sn 2+ 、Hg + 、Fe 2+ , K + In the presence of Sn 2+ 、Cd 2+ 、Zn 2+ 、Cu + 、Ag + In the presence of Hg + 、Sn 2+The blue fluorescence was significantly enhanced in the presence of

[0125] Soft salt SA in Hg + 、Fe 3+ Green fluorescence was significantly quenched in the presence of Sn 2+ In the presence of Cu + 、Zn 2+ 、Ag + In the presence of Cu + 、Ag + 、Zn 2+ In the presence of Sn 2+ In the presence of Hg + 、Cu + 、Fe 3+ Green fluorescence was significantly quenched in the presence of Sn 2+ 、Ag + 、Zn 2+ The red fluorescence was significantly enhanced in the presence of

[0126] 3. Filter paper test of complex 2a, phosphorescent soft salt complexes SB and SD

[0127] Weigh 1 mg of phosphorescent complex 2a, SB, and SD, add them to dimethyl sulfoxide to prepare 20 mmol / L solution, and then dilute it to 100 μmol / L solution. 2+ 、Cu + 、Ag + Prepare ion solutions with final concentrations of 2.5mmol / L, 5mmol / L, and 10mmol / L. Cut the filter paper into rectangles of the same size. Add the solutions of 2a, SB, and SD onto the filter paper respectively. After drying in an oven, add the prepared ion solutions respectively. The results are as follows: Figure 15 As shown;

[0128] It can be seen from the figure that the filter paper with soft salt SB has no color under natural light and fluorescence before adding ions (2.5, 5, 10mM). + After ionization, there is no color change in natural light, but the color of the filter paper changes under fluorescence, and the color turns orange; the filter paper with soft salt SD, plus Ag + Before ions, natural light is colorless and fluorescence is yellow-green; when Ag is added + After the addition of ions, the natural light filter paper turns light red and the fluorescence turns orange-red.2+ Before ionization, the filter paper is colorless under both natural light and fluorescence. After ion addition, there is no color change under natural light, but under fluorescence, the filter paper changes color to a light red. Therefore, test paper with the complex can be used for metal ion detection. The naked eye can directly determine the presence of ions, making this method more convenient and quicker.

[0129] 4. Zn in complex 2a, phosphorescent soft salt complexes SB and SD 2+ 、Ag + Fluorescence titration

[0130] Weigh 1 mg each of complex 2a, soft salt SB, and SD and add them to dimethyl sulfoxide to prepare a 20 mmol / L stock solution. Take 3 μL into a 5 mL centrifuge tube, add 27 μL of dimethyl sulfoxide to each tube, and then add water to prepare 40 μmol / L and 20 μmol / L solutions, respectively. Add a series of Zn 2+ 、Ag + Ion solution, under 400nm excitation, obtain its fluorescence emission spectrum, the results are as follows Figure 16 As shown;

[0131] It can be seen from the figure that Zn 2+ Titration of soft salt SB, with the increase of metal ion concentration, the fluorescence intensity of soft salt SB at 573nm increased by about 9 times. + Titration of soft salt SD, with the increase of metal ion concentration, at 556nm, when the metal concentration is 80μM, the fluorescence intensity is the strongest, increasing by about 6 times. + When the complex 2a was titrated, as the metal ion concentration increased, the fluorescence intensity at 555 nm reached its highest value when the metal concentration was 500 μM, increasing by about 3 times.

[0132] The calculation shows that the complex 2a has a strong affinity for Zn 2+ The detection limit of soft salt B for Zn 2+ The detection limit of soft salt D for Ag is 9.4 μM. + The detection limit was 362 nM.

[0133] In summary, the phosphorescent metal complex prepared in the present invention can be used in the detection of anions and cations with significant effects.

[0134] Example 10: Application of phosphorescent metal complexes in cell imaging

[0135] 1. AIE effect detection of complexes 1a, 1b, 2a, 2b and phosphorescent soft salt complexes SA, SB, SC, SD

[0136] Weigh 1 mg each of complexes 1a, 1b, 2a, 2b and soft salts SA, SB, SC, and SD, add them to dimethyl sulfoxide to prepare a 20 mmol / L mother solution, take 3 μL of the mother solution into a 5 mL centrifuge tube, add 27 μL of dimethyl sulfoxide to each tube, and then add a mixture of water and dimethyl sulfoxide (1:9, 3:7, 5:5, 7:3, 9:1) respectively. The above solutions are measured for fluorescence under 320 nm / 400 nm excitation light to obtain fluorescence spectra. The results are shown in Figure 2. Figure 17-19 ;

[0137] It can be seen from the figure that in the mixed solution of DMSO and water, the fluorescence intensity of complex 1a is the highest when the water content increases, and the green fluorescence intensity at 523nm increases by about 4 times at 70% water content (7:3); in the solution of complex 1b, the fluorescence intensity gradually decreases with the increase of water content, and the fluorescence is the lowest when the water content is 90% (9:1), and the blue fluorescence intensity at 411nm decreases by about 5 times; in the solution of complex 2a, the fluorescence is the highest when the water content increases, and the green fluorescence intensity at 487nm increases by about 3 times at 90% water content (9:1); in the solution of complex 2b, the fluorescence is the lowest when the water content is 90% (9:1), and the blue fluorescence intensity at 423nm decreases by about 4 times.

[0138] In a mixed solution of DMSO and water, soft salt A showed a maximum fluorescence intensity at 90% water content (9:1) with increasing water content, and the green fluorescence intensity at 513 nm increased by about 6 times; in soft salt B solution, the fluorescence gradually red-shifted with increasing water content, and reached a maximum fluorescence at 90% water content (9:1), and the red fluorescence intensity at 576 nm increased by about 50 times; in soft salt C solution, the fluorescence reached a maximum fluorescence at 50% water content (5:5) with increasing water content, and the red fluorescence intensity at 538 nm increased by about 123 times; in soft salt D solution, the fluorescence reached a maximum fluorescence at 70% water content (7:3) with increasing water content, and the green fluorescence intensity at 515 nm increased by about 7 times;

[0139] In summary, complexes 1a, 2a, and the four soft salts all exhibit aggregation-induced emission effects and have strong resistance to photobleaching in the aggregated state, and are expected to become good bioimaging materials.

[0140] 2. Effects of glutathione on complexes 1a, 1b, 2a, 2b and phosphorescent soft salt complexes SA, SB, SC, SD

[0141] Weigh 1 mg each of complexes 1a, 1b, 2a, 2b and soft salts SA, SB, SC, and SD, add them to dimethyl sulfoxide to prepare a 20 mmol / l stock solution, take 3 μL of the stock solution into a 5 mL centrifuge tube, add 27 μL of dimethyl sulfoxide to each tube, dilute with water to 20 μmol / L or 40 μmol / L, then add GSH, respectively, to a final concentration of 5 mmol / L. After mixing at room temperature for 0, 10, 30, 60, and 120 minutes, excite at 320 nm / 400 nm to obtain its fluorescence emission spectrum. The results are as follows. Figure 20-23 As shown;

[0142] Complexes 1a, 1b, and 2a showed no significant fluorescence changes in the presence of glutathione over time. Complex 2b exhibited a 17-fold increase in blue fluorescence intensity at 361 nm over time in the presence of glutathione, with the intensity increasing by approximately 17 times at 120 minutes. This suggests that 1a, 1b, and 2a are less stable than glutathione, indicating that 2b exhibits poor stability.

[0143] Soft salts SA and SB showed no significant fluorescence changes in the presence of glutathione at different times. However, in the presence of glutathione, the blue fluorescence intensity of soft salt C at 361 nm increased approximately sevenfold over time, at 120 minutes. In the presence of glutathione, the green fluorescence of soft salt D shifted red from 509 nm to 566 nm over time, with the peak red-shifting by 57 nm. This suggests that soft salts SA and SB are less likely to bind to glutathione and have good stability, while SC is less stable. However, after binding to glutathione, the fluorescence intensity of SD remains unchanged but shifts red, enhancing its penetration.

[0144] 3. Imaging of phosphorescent soft salt complexes in tumor cells

[0145] HeLa cells (human cervical cancer cell line) in good growth condition were digested with trypsin and inoculated into confocal culture dishes. They were cultured in an incubator containing 5% CO2 at 37°C. When the density of HeLa cells reached 70%, soft salt SB was added. The concentration of SB for incubating cells was 40 μmol / L. The cells in the dish were cultured for 24 hours. After incubation for 10-15 minutes, the culture medium was removed, and the cells were washed twice with PBS. The cells were immediately observed with a laser confocal microscope. The results are shown in Fig. Figure 24 Under excitation conditions of different wavelengths, soft salt SB can have dual emission of green and red fluorescence in cells, and the two fluorescences are not well co-localized, which means that the soft salt molecules emit two different colors of light in the cells. After incubation for 6 hours, the two phosphorescence intensities of soft salt SB are significantly enhanced, and the degree of co-localization of green and red fluorescence increases. The experiment proves that our soft salt material can be used for imaging and tracing of red and green fluorescence probes in living cells in the future.

[0146] Example 11: Solvent Effects of Phosphorescent Soft Salt Complexes SA, SB, SC, and SD

[0147] Weigh 1 mg each of soft salts SA, SC, and SD, add them to dimethyl sulfoxide to prepare a 20 mmol / L stock solution. Weigh 1 mg of soft salt SB and add it to dimethyl sulfoxide to prepare a 40 mmol / L stock solution. Take 3 μL into a 5 mL centrifuge tube, add 27 μL of dimethyl sulfoxide to each tube, and then add water, toluene, N,N'-dimethylformamide, cyclohexane, dimethyl sulfoxide, ethanol, methanol, ethyl acetate, acetonitrile, acetone, ether, chloroform, and dichloromethane to prepare a 20 μmol / L or 40 μmol / L solution. Under 320 / 400 nm excitation, obtain its fluorescence emission spectrum. The results are as follows. Figures 25-26 As shown;

[0148] Compared with aqueous solutions, the green fluorescence of soft salt SA is enhanced in ethanol, N,N'-dimethylformamide, methanol, and acetone, and weakened in acetonitrile, toluene, dimethyl sulfoxide, chloroform, ethyl acetate, diethyl ether, and cyclohexane. Soft salt SB emits blue light in toluene, N,N'-dimethylformamide, cyclohexane, and dimethyl sulfoxide, with the strongest blue light emission in dimethyl sulfoxide. Soft salt SC has enhanced blue fluorescence in toluene, diethyl ether, N,N'-dimethylformamide, cyclohexane, and dichloromethane. Soft salt SD emits blue fluorescence in dimethyl sulfoxide and cyclohexane, and emits red fluorescence in dichloromethane, ethyl acetate, diethyl ether, and toluene. This shows that the four soft salts can also be used to detect different solvents.

[0149] Example 12: Application of phosphorescent complexes in anti-tumor activity

[0150] 1. Anti-tumor activity detection of phosphorescent complexes

[0151] The cytotoxicity of the complexes prepared in Examples 1-8 to HeLa (human cervical cancer cell line) was determined, with cisplatin (Cis) as the positive control group, and the specific method was as follows:

[0152] 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 drugs at different concentrations, incubated in a 5% CO2, 37°C incubator for 48 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. The OD value was measured using a microplate reader with the wavelength set to 492 nm;

[0153] Calculate the survival rate of the tested tumor cells, plot and calculate IC50 The anti-tumor activity of the complexes was evaluated by quantifying the toxicity of the complexes. The results are shown in the table below. Compared with cisplatin, the anti-tumor activity of complexes 1a-2b was weaker than that of cisplatin. After interaction with each other, except for the soft salt SC which had no anti-tumor activity, the anti-tumor toxicity of soft salt SA was stronger than that of cisplatin, while SB and SD were slightly weaker than cisplatin.

[0154]

[0155] 2. Effect of amino acids on phosphorescent soft salt complexes SA, SB, SC, and SD

[0156] Weigh 1 mg each of soft salts SA, SB, SC, and SD, add them to dimethyl sulfoxide to prepare a 20 mmol / L stock solution, take 3 μL of the stock solution into a 5 mL centrifuge tube, add 27 μL of dimethyl sulfoxide to each tube, dilute with water to 20 μmol / L or 40 μmol / L, add L-methionine, L-proline, L-tryptophan, phenylalanine, L-aspartic acid, L-histidine, L-glutamic acid, L-cysteine ​​and other amino acids, and the final concentration of the amino acids is 5 mmol / L. Measure the fluorescence of the above solution under 320 nm / 400 nm excitation light to obtain the fluorescence spectrum. The results are shown in Figure 2. Figures 27-28 ;

[0157] Amino acids have no effect on the luminescence of soft salts SA, SB, and SD, while histidine itself emits blue light. Tryptophan increases the blue fluorescence intensity of soft salt SC at 361 nm by about 3 times.

[0158] 3. Effect of solution pH on the luminescence of phosphorescent soft salt complexes SA, SB, SC, and SD

[0159] Weigh 1 mg each of soft salts SA, SB, SC, and SD, add them to dimethyl sulfoxide to prepare a 20 mmol / L stock solution. Take 3 μL of the stock solution into a 5 mL centrifuge tube, add 27 μL of dimethyl sulfoxide to each tube, and dilute to 20 μmol / L or 40 μmol / L with sodium hydrogen phosphate-citric acid buffer solutions of pH 5.8, 6.2, 6.6, 7.0, and 7.4. Measure the fluorescence of the above solutions under 320 nm / 400 nm excitation light to obtain fluorescence spectra. The results are shown in Figures 29-30.

[0160] The change of solution pH had no effect on the luminescence of soft salts SA, SB, SC and SD.

Claims

1. A phosphorescent metal complex having any of the following structural formulas: 、 、 、 。 2. Application of a phosphorescent metal complex in anion detection, characterized in that: For quenching detection of NO2 - , ClO - , I - ; For enhanced detection of HSO3 - , ClO - ; For enhanced detection of CH3COO - , ClO - , I - 、NO3 - 、Cl - 、ClO4 - 、SO4 2- 、H2PO4 - ; For enhanced detection of ClO - 、HSO3 - ; For quenching detection of NO2 - , ClO - 、HSO3 - , I - ; For quenching detection of CO3 2- , ClO - ; For enhanced detection of ClO - 、HCO3 - ; For quenching detection of NO2 - , ClO - 、HSO3 - , I - .

3. Application of a phosphorescent metal complex in cation detection, characterized in that: For quenching detection of Hg + 、Cu + 、Fe 3+ ; For enhanced detection of Sn 2+ 、Hg + 、Fe 2+ , K + ; For enhanced detection of Sn 2+ 、Cd 2+ 、Cu + 、Ag + 、Zn 2+ ; For enhanced detection of Hg + 、Sn 2+ ; For quenching detection of Hg + 、Fe 3+ ; For enhanced detection of Cu + 、Zn 2+ 、Ag + ; For enhanced detection of Cu + 、Ag + 、Zn 2+ ; For quenching detection of Hg + 、Fe 3+ 、Cu + , enhanced detection Sn 2+ 、Zn 2+ 、Ag + .

4. Use of a phosphorescent metal complex in the preparation of a cell diagnosis and treatment imaging reagent, characterized in that: The complex has aggregation-induced emission effect; The phosphorescent metal complex is 、 、 、 .

5. Use of a phosphorescent metal complex in the preparation of an anti-human cervical cancer drug, characterized in that: The phosphorescent metal complex is 、 、 、 、 、 .

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