Triphenylamine derivative, aggregation-induced emission material as well as preparation method and application of aggregation-induced emission material
By using a tripaniline derivative containing aromatic glycoside groups as the aggregation-induced luminescent material, the problem of inadvertent and insufficient sensitivity for β-galactosidase detection in the prior art is solved, and a high sensitivity and selectivity detection effect is achieved.
Patent Information
- Application Number
- CN202510034558.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-09
- Publication Date
- 2025-04-29
AI Technical Summary
The existing aggregation-induced luminescent materials are not targeted for the detection of β-galactosidase, and the detection effect is not sensitive.
A triphenylamine derivative is used as an aggregation-induced luminescent material, whose structure contains aromatic glycoside groups. This material is prepared by a specific preparation method for detection of β-galactosidase.
The tripaniline derivative has excellent optical properties and stability, significantly improves the detection sensitivity and selectivity of β-galactosidase, and is suitable for detection in biological organisms.
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Figure CN119899224A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of aggregation-induced emission materials, and in particular to a triphenylamine derivative, an aggregation-induced emission material, and a preparation method and application thereof. Background Art
[0002] Since the phenomenon of aggregation-induced emission (AIE) was first proposed in 2001, scientists have promoted the rapid development of AIE molecular materials by designing specific molecular structures, especially introducing functional groups that can promote molecular stacking and restrict free rotation. Materials with piezoluminescent properties such as triphenylamine derivatives and tetraphenylethylene derivatives have been widely studied. These materials show high fluorescence efficiency in the aggregated state, further broadening their application prospects in various sensing and display devices.
[0003] β-galactosidase is a lysosomal hydrolase that plays an important role in the catalytic hydrolysis of glycosidic bonds. The detection of β-galactosidase activity in vivo based on fluorescence sensors has made significant progress in the past few years, especially the use of aggregation-induced emission technology, which has improved background minimization and cell permeability, and enhanced the sensitivity, selectivity and spatiotemporal resolution of detection. Aggregation-induced emission not only provides a basis for AIE-based cationic sensors, but also provides new possibilities for the detection of enzyme activity in vivo based on fluorescence sensors.
[0004] However, existing aggregation-induced luminescence materials have some shortcomings in the detection of β-galactosidase. For example, most aggregation-induced luminescence materials do not have targeted detection of β-galactosidase, and the detection sensitivity is not high.
[0005] Therefore, there is an urgent need for a new galactose-based aggregation-induced emission material for the detection of β-galactosidase in vivo. Summary of the invention
[0006] The purpose of the present invention is to overcome the above technical deficiencies and provide a method for solving the technical problem of how to detect β-galactosidase in the prior art.
[0007] In order to achieve the above technical purpose, the technical solution of the present invention provides a triphenylamine derivative, whose structural formula is:
[0008] In addition, the present invention also provides a method for preparing the above triphenylamine derivative, comprising the following steps: dissolving compound IV in water; Yudi After adding a solvent, add a base to adjust the pH value to 9-10, stir and react under argon protection, then add an acidic resin to the reaction solution, adjust the pH to 4-5 and continue stirring to obtain the triphenylamine derivative; the structural formula of the compound IV is
[0009]
[0010] In any embodiment, the first solvent is a mixed solution of dichloromethane and anhydrous methanol; and / or the base is sodium methoxide; and / or the stirring reaction time under argon protection is 1 to 5 hours.
[0011] In any embodiment, the process further comprises preparing compound IV before preparing the triphenylamine derivative:
[0012] Compound III and galactose bromide are dissolved in a second solvent, a promoter is added, argon is introduced, and the mixture is stirred to react to obtain compound IV; the structural formula of compound III is
[0013]
[0014] In any embodiment, the promoter is cesium carbonate; the molar ratio of cesium carbonate to compound III is (1-3):1; and / or, the stirring reaction time after the introduction of argon is 5-12 hours; and / or, the second solvent is acetonitrile.
[0015] In any embodiment, the process further comprises preparing compound III before preparing compound IV:
[0016] Compound II is dissolved in a third solvent, cooled to -15 to -5°C, and boron tribromide is added dropwise and stirred to react to obtain compound III; the structural formula of compound II is The third solvent is dichloromethane; and / or the molar ratio of the boron tribromide to the compound II is (1-3):1; and / or the stirring reaction time after the boron tribromide is added dropwise is 3-12 hours.
[0017] In office In an embodiment, the preparation of compound II is further included before the preparation of compound III: triphenylamine and p-methoxybenzoyl chloride are dissolved in a fourth solvent, and a Lewis acid is added and stirred to react to obtain compound II; the fourth solvent is dichloromethane; and / or the Lewis acid is aluminum chloride; the molar ratio of the Lewis acid to the triphenylamine is (1 to 1.5):1; the stirring reaction time after the Lewis acid is added is 3 to 12 hours.
[0018] In addition, the present invention also provides an aggregation-induced emission material, comprising the above triphenylamine derivative or the triphenylamine derivative prepared by the above preparation method.
[0019] In addition, the present invention also proposes the use of a triphenylamine derivative or the triphenylamine derivative prepared by the above preparation method as an aggregation-induced emission material in the detection of β-galactosidase.
[0020] In any embodiment, the detection temperature of β-galactosidase is 0-40° C.; and / or the detection pH value ranges from 4-9.
[0021] Compared with the prior art, the beneficial effects of the present invention include: the triphenylamine derivative proposed by the present invention has an aromatic glycoside group, excellent optical properties, stable properties, and can be used as an aggregation-induced luminescence material for the detection of β-galactosidase. BRIEF DESCRIPTION OF THE DRAWINGS
[0022] Figure 1 is the hydrogen nuclear magnetic resonance spectrum of compound II prepared in Example 1;
[0023] Figure 2 is the carbon NMR spectrum of compound II prepared in Example 1;
[0024] Figure 3 is a high-resolution mass spectrum of compound II prepared in Example 1;
[0025] Figure 4 This is the fluorescence spectrum of Example 2 when different β-galactosidases were added in PBS:DMSO=1:1.
[0026] Figure 5 This is the fluorescence spectrum of Example 2 when different β-galactosidases were added in PBS:DMSO=1:9.
[0027] Figure 6 This is the fluorescence spectrum of Example 2 when different β-galactosidases were added in PBS:DMSO=9:1. DETAILED DESCRIPTION
[0028] The "range" disclosed in the present application is defined in the form of a lower limit and an upper limit, and a given range is defined by selecting a lower limit and an upper limit, and the selected lower limit and upper limit define the boundaries of the particular range. The range defined in this way can be inclusive or exclusive of the end values, and can be arbitrarily combined, that is, any lower limit can be combined with any upper limit to form a range. For example, if a range of 60 to 120 and 80 to 110 is listed for a particular parameter, it is understood that a range of 60 to 110 and 80 to 120 is also expected. In addition, if the minimum range values 1 and 2 are listed, and if the maximum range values 3, 4 and 5 are listed, the following ranges can all be expected: 1 to 3, 1 to 4, 1 to 5, 2 to 3, 2 to 4 and 2 to 5. In the present application, unless otherwise specified, the numerical range "a to b" represents an abbreviation of any real number combination between a and b, where a and b are both real numbers. For example, the numerical range "0-5" means that all real numbers between "0-5" are listed in this document, and "0-5" is just an abbreviation of these numerical combinations. In addition, when a parameter is expressed as an integer ≥ 2, it is equivalent to disclosing that the parameter is, for example, an integer of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, etc.
[0029] If there is no special explanation, the "include" and "comprising" mentioned in this application are open-ended or closed-ended. For example, the "include" and "comprising" may mean that other components not listed may also be included or only the listed components may be included or only the listed components may be included.
[0030] If not specifically stated, in this application, the term "or" is inclusive. For example, the phrase "A or B" means "A, B, or both A and B". More specifically, any of the following conditions satisfies the condition "A or B": A is true (or exists) and B is false (or does not exist); A is false (or does not exist) and B is true (or exists); or both A and B are true (or exist).
[0031] This specific embodiment provides a triphenylamine derivative, which has a chemical formula of C 31 H 29 NO7, its structural formula is:
[0032] This specific embodiment also proposes a method for preparing a triphenylamine derivative, comprising the following steps:
[0033] S1, dissolving triphenylamine and p-methoxybenzoyl chloride in a fourth solvent, dichloromethane, adding Lewis acid aluminum chloride and stirring to react for 3 to 12 hours; after the reaction, adding ice water, extracting with dichloromethane, concentrating the organic phase and purifying it with a thin layer chromatography silica gel column to obtain a white powder solid compound II; the molar ratio of the Lewis acid to the triphenylamine is (1 to 1.5):1;
[0034] S2, dissolving compound II in a third solvent, dichloromethane, cooling to -15 to -5°C, adding boron tribromide dropwise, stirring and reacting for 3-12 hours, after the reaction, cooling the reaction system with an ice-water bath, adding water dropwise to quench the reaction, filtering, and placing the filter cake in a vacuum drying oven to obtain a yellow-brown powdery solid compound III; the structural formula of the compound II is The molar ratio of the boron tribromide to the compound II is (1-3):1;
[0035] S3, dissolving compound III and galactose bromide in the second solvent acetonitrile, adding a promoter, passing argon gas and stirring to react for 5 to 12 hours; after the reaction, concentrating the solvent, extracting with CH2Cl2, and purifying the obtained crude product by thin layer chromatography on a silica gel column to obtain a yellow-green solid compound IV; the structural formula of the compound III is The promoter is cesium carbonate; the molar ratio of cesium carbonate to compound III is (1-3):1;
[0036] After compound IV is dissolved in the first solvent, sodium methoxide is added to adjust the pH value to 9-10, and the reaction is stirred for 1-5 hours under argon protection. Then, an acidic resin is added to the reaction solution, the pH is adjusted to 4-5, stirring is continued, and suction is filtered. The filtrate is spin-dried to obtain the triphenylamine derivative; the structural formula of compound IV is The first solvent is a mixed solution of dichloromethane and anhydrous methanol.
[0037] The relevant reaction formula is as follows:
[0038]
[0039] The method for preparing the aromatic glycoside triphenylamine derivative proposed in this specific embodiment uses triphenylamine and p-methoxybenzene as starting materials and is prepared through four steps of Friedel-Crafts acylation, demethoxylation, glycosylation, and deacetylation. It has the characteristics of simple operation, innovative method, and good substrate compatibility.
[0040] This specific embodiment also provides an aggregation-induced emission material, including the above triphenylamine derivative or the triphenylamine derivative prepared by the above preparation method.
[0041] This specific embodiment also proposes the use of the above triphenylamine derivative or the triphenylamine derivative prepared by the above preparation method as an aggregation-induced emission material in the detection of β-galactosidase; the detection temperature of β-galactosidase is 0-40°C; the detection pH value ranges from 4 to 9.
[0042] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.
[0043] In the present invention, reference is made to “some embodiments”, “this embodiment”, examples, etc., which describe a subset of all possible embodiments, but it can be understood that “some embodiments” may be the same subset or different subsets of all possible embodiments and may be combined with each other without conflict.
[0044] If similar descriptions of "first / second" appear in the application documents, the following instructions are added. In the following description, the terms "first\second\third" involved are merely used to distinguish similar objects and do not represent a specific ordering of the objects. It can be understood that "first\second\third" can be interchanged in a specific order or sequence where permitted, so that the embodiments described herein can be implemented in an order other than that illustrated or described herein.
[0045] In this embodiment, the term "and / or" is merely a description of the association relationship between associated objects, indicating that three relationships may exist. For example, object A and / or object B may represent three situations: object A exists alone, object A and object B exist at the same time, and object B exists alone.
[0046] Hereinafter, the embodiments of the present application will be described. The embodiments described below are exemplary and are only used to explain the present application, and should not be construed as limiting the present application. If no specific techniques or conditions are indicated in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are used. If the manufacturer is not indicated in the reagents or instruments used, they are all conventional products that can be obtained commercially.
[0047] The acidic resin in the following examples is cation exchange resin IR120 (sodium type), Amberlite IR120, CAS No.: 9002-23-7, manufacturer: Adamas.
[0048] Example 1
[0049] This embodiment provides a triphenylamine derivative, which has a chemical formula of C 31 H 29 NO7, the structural formula is The preparation method of the triphenylamine derivative is as follows:
[0050] (1) Preparation of Compound II
[0051] Add triphenylamine (2.4514 g, 10 mmol, 1 eq) and p-methoxybenzoyl chloride (1.3623 mL, 10 mmol, 1 eq) to a dry reaction bottle, then add 10 mL of CH2Cl2, then add aluminum chloride (1.3324 g, 10 mmol, 1 eq) and 5 mL of CH2Cl2, and stir magnetically at room temperature.
[0052] The reaction was complete after monitoring by TLC plate. The mixture in the reaction flask was poured into a separatory funnel, 50 mL of ice water was added to the separatory funnel, and then 10 mL of CH2Cl2 was used for extraction. The organic phase was separated, dried over anhydrous Na2SO4, and the organic phase was concentrated to finally obtain a crude product. The crude product was purified by thin layer chromatography on a silica gel column to obtain a white powder solid compound II with a purity of 99.2%. The yield was 2.8843 g, and the yield was 76.01%. The hydrogen spectrum and carbon spectrum data of compound II are as follows:
[0053] 1 H NMR(400MHz,Chloroform-d)δ7.85–7.78(m,2H),7.71–7.65(m,2H),7.38–7.26(m,4H),7.19(d,J=1.4H z,2H),7.17(d,J=1.1Hz,2H),7.15–7.09(m,2H),7.03(d,J=8.8Hz,2H),6.99–6.92(m,2H),3.88(s,3H).
[0054] 13 C NMR(101MHz,Chloroform-d)δ194.21,162.76,151.57,146.69,132.33,132. 18,131.70,130.92,130.38,129.61,125.87,124.48,119.88,113.45,55.47.
[0055] (2) Preparation of Compound III
[0056] Add compound II (1.0304 g, 2.64 mmol, 1 eq) to the reaction flask, evacuate it, fill it with argon, repeat 3 times. Add 10 mL of CH2Cl2 to the reaction flask, cool it to -15°C, then slowly drop 2 mL of boron tribromide solution, and stir at room temperature after the dropwise addition is complete.
[0057] The reaction was complete after monitoring by TLC plate. After cooling the reaction bottle with ethanol at -15°C, 3-5 drops of water were slowly added to quench the reaction, and then 5 mL of water was added, filtered, and the filter cake was dried in a vacuum oven at 40°C to finally obtain a yellow-brown powdery solid compound III with a purity of 98.1%. The yield was 0.7479 g and the yield was 75.43%. The hydrogen spectrum and carbon spectrum data of compound III are as follows:
[0058] 1 H NMR(400MHz,Chloroform-d)δ7.75(d,J=8.7Hz,2H),7.67(d,J=8.8Hz,2H),7.36–7.28(m,4H),7. 17(d,J=7.1Hz,4H),7.18–7.09(m,2H),7.01(d,J=8.8Hz,2H),6.90(d,J=8.6Hz,2H),6.62(s,1H).
[0059] 13 C NMR (101MHz, Chloroform-d) δ194.48,151.68,132.50,131.77,130.85,130.14,129.60,125.91,124.53,119.76,115.05.
[0060] (3) Preparation of Compound IV
[0061] Galactoside (0.1001 g, 0.29 mmol, 1.4 eq), compound IV (0.0753 g, 0.21 mmol, 1 eq) and cesium carbonate (0.1358 g, 0.42 mmol, 2 eq) were dissolved in 5 mL of CH3CN, introduced with argon, and stirred at room temperature.
[0062] The reaction was complete after monitoring by TLC plate. The reaction solution was concentrated by rotary evaporator, and after removing the solvent, it was extracted with CH2Cl2, and the organic phase was dried with anhydrous Na2SO4 and concentrated again to obtain a crude product. The crude product was purified by thin layer chromatography on silica gel column to obtain yellow-green solid compound IV with a purity of 99.5%. The yield was 0.0860g and the yield was 60.00%. The hydrogen spectrum and carbon spectrum data of compound IV are as follows:
[0063] 1H NMR(400MHz,Chloroform-d)δ7.74(d,J=8.8Hz,2H),7.62(d,J=8.8Hz,2H),7.32–7.20(m,4H),7.14(s,2H),7.13–7.05(m,4H),6.99(dd,J=20.6,8.8Hz ,4H),5.41–5.47(m,1H),5.09(d,J=3.4Hz,1H),4.21–4.16(m,1H),4.14(d, J=6.0Hz,1H),4.11–4.09(m,1H),4.06(t,J=7.1Hz,2H),2.03–1.98(m,12H).
[0064] 13 C NMR(101MHz,Chloroform-d)δ193.93,171.17,170.37,170.22,170.11,169.38,159.47,151.85,146.50,133.36,131.87,131.79 ,129.63,125.97,124.66,119.57,116.02,98.91,71.22,70.76,68.52,66.83,61.37,60.40,21.05,20.74,20.68,20.66,20.59.
[0065] HRMS(ESI):calcd for[C 39 H 37 NO 11 +H] + m / z 696.2439,found m / z696.2433.
[0066] (4) Preparation of Compound I
[0067] Compound IV (0.0614 g, 0.08 mmol, 1 eq) was dissolved in a mixed solution of dichloromethane / anhydrous methanol (3 mL / 9 mL), and 0.03 g of sodium methoxide was added to adjust the pH to 9-10, and the mixture was stirred at room temperature under argon protection.
[0068] The reaction was complete after monitoring by TLC plate. 2.7 g of acidic resin was added to the reaction solution, pH was adjusted to 5, stirred at room temperature for 10 min, then filtered, and the filtrate was spin-dried. Finally, a yellow-green solid compound I was obtained with a purity of 99.0%. The yield was 0.0372 g, and the yield was 79.90%. The hydrogen spectrum and carbon spectrum data of compound I are as follows:
[0069] 1H NMR (400MHz, Methanol-d4) δ7.71(d,J=8.4Hz,2H),7.64(s,1H),7.61(d,J=8.5Hz,1H),7.33(t,J=7.8Hz,4H) ,7.16(dd,J=17.6,9.8Hz,9H),6.94(d,J=8.5Hz,2H),3.82–3.74(m,3H),3.72(s,1H),3.31(d,J=15.9Hz,4H).
[0070] 13 C NMR (101MHz, Methanol-d4) δ196.40,170.43,132.94,130.83,127.27,126.06,120.17,117.15,102.25,77.06,72.11,70.18,62.38,48.74,30.72.
[0071] HRMS(ESI):calcd for[C 31 H 29 NO7+H] + m / z 528.2017,found m / z528.2012.
[0072] from Figure 1 , Figure 2 The NMR spectroscopy data and Figure 3 It can be judged from the high-resolution mass spectrometry data that the final product synthesized by the present invention is indeed compound I.
[0073] The reaction formula is as follows:
[0074]
[0075] Embodiment 2:
[0076] Application of aromatic glycoside triphenylamine derivatives as aggregation-induced emission materials
[0077] Considering the solubility of sugar compounds, a mixed solvent of phosphate buffer solution (PBS) and dimethyl sulfoxide (DMSO) was used.
[0078] (1) The solvent volume ratio is PBS:DMSO = 1:1
[0079] Five sample bottles were taken, compound I (10 uM) was added respectively, and after adding PBS / DMSO mixed solvent, 0, 25 U, 50 U, 100 U, and 200 U of β-galactosidase were added in sequence. After standing for 1 hour, the fluorescence spectrum was tested.
[0080] Figure 4The fluorescence spectra of different β-galactosidase added in PBS:DMSO=1:1 are shown. The fluorescence intensity increases after adding β-galactosidase in the range of 25U to 200U. The corresponding wavelength of the maximum fluorescence intensity before and after adding β-galactosidase is 470nm.
[0081] (2) The solvent volume ratio is PBS:DMSO = 1:9
[0082] Three sample bottles were taken, compound I (10 uM) was added respectively, and after adding PBS / DMSO mixed solvent, 0, 100 U, and 200 U of β-galactosidase were added in sequence. After standing for 1 hour, the fluorescence spectrum was tested.
[0083] Figure 5 The fluorescence spectra of different β-galactosidases added in PBS:DMSO=1:9 are shown. In the solvent of PBS:DMSO=1:9, the addition of β-galactosidase increases the fluorescence intensity, and increases with the increase of the amount of β-galactosidase added.
[0084] (3) The solvent volume ratio is PBS:DMSO = 9:1
[0085] Four sample bottles were taken, compound I (10 uM) was added respectively, and after adding PBS / DMSO mixed solvent, 0, 5.4, 8.1, and 10.8 U / mL of β-galactosidase were added in sequence. After standing for 1 hour, the fluorescence spectrum was tested.
[0086] Figure 6 The fluorescence spectra of different β-galactosidases added in PBS:DMSO=9:1 are shown. In PBS:DMSO=9:1 solvent, the fluorescence intensity increases with the increase of the concentration of β-galactosidase added.
[0087] The above experimental results show that aromatic glycoside triphenylamine derivatives have excellent optical properties and good stability, and can be used as aggregation-induced emission materials for the detection of β-galactosidase, laying the foundation for the promotion of this type of material.
[0088] The specific implementation of the present invention described above does not constitute a limitation on the protection scope of the present invention. Any other corresponding changes and modifications made based on the technical concept of the present invention should be included in the protection scope of the claims of the present invention.
Claims
1. A triphenylamine derivative, characterized in that Its structural formula is:
2. A method for preparing the triphenylamine derivative according to claim 1, characterized in that: The following steps are involved: Combination Material IV After being dissolved in the first solvent, a base is added to adjust the pH value to 9-10, and the reaction is stirred under argon protection, and then an acidic resin is added to the reaction solution, and the pH is adjusted to 4-5 and continued to stir to obtain the triphenylamine derivative; the structural formula of the compound IV is 3. The method for preparing a triphenylamine derivative according to claim 2, characterized in that: The first solvent is a mixed solution of dichloromethane and anhydrous methanol; and / or the base is sodium methoxide; and / or the stirring reaction time under argon protection is 1 to 5 hours.
4. The method for preparing a triphenylamine derivative according to claim 2, characterized in that: In making three Aniline derivatives Before It also includes the preparation of compound IV: Compound III and galactose bromide are dissolved in a second solvent, a promoter is added, argon is introduced, and the mixture is stirred to react to obtain compound IV; the structural formula of compound III is 5. The method for preparing a triphenylamine derivative according to claim 4, characterized in that: The promoter is cesium carbonate; the molar ratio of cesium carbonate to compound III is (1-3):1; and / or, the stirring reaction time after the introduction of argon is 5-12 hours; and / or, the second solvent is acetonitrile.
6. The method for preparing a triphenylamine derivative according to claim 4, characterized in that: Before preparing compound IV, the method further includes preparing compound III: Compound II is dissolved in a third solvent, cooled to -15 to -5°C, and boron tribromide is added dropwise and stirred to react to obtain compound III; the structural formula of compound II is The third solvent is dichloromethane; and / or the molar ratio of the boron tribromide to the compound II is (1-3):1; and / or the stirring reaction time after the boron tribromide is added dropwise is 3-12 hours.
7. root The method for preparing a triphenylamine derivative according to claim 6, characterized in that: Before preparing compound III, the method also includes preparing compound II: dissolving triphenylamine and p-methoxybenzoyl chloride in a fourth solvent, adding Lewis acid and stirring to react to obtain compound II; the fourth solvent is dichloromethane; and / or the Lewis acid is aluminum trichloride; the molar ratio of the Lewis acid to the triphenylamine is (1-1.5):1; and / or the stirring reaction time after adding the Lewis acid is 3-12 hours.
8. An aggregation-induced emission material, characterized in that: The invention comprises the triphenylamine derivative according to claim 1 or the triphenylamine derivative prepared by the preparation method according to any one of claims 2 to 7.
9. Use of the triphenylamine derivative according to claim 1 or the triphenylamine derivative prepared by the preparation method according to any one of claims 2 to 8 as an aggregation-induced emission material in the detection of β-galactosidase.
10. The use according to claim 9, characterized in that: The detection temperature of β-galactosidase is 0-40°C; and / or the detection pH value ranges from 4-9.