Tetraphenyl ethylene derivative TPE-1s crystal probe as well as preparation method and application thereof
By preparing a tetraphenylethylene derivative TPE-1s crystal probe with a monoclinic crystal structure, the problem of insufficient selectivity and sensitivity of tetraphenylethylene derivatives in the detection of cysteine in the existing technology is solved, and a highly selective and sensitive detection effect is achieved, which is suitable for biological imaging applications.
Patent Information
- Application Number
- CN202510775073.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-11
- Publication Date
- 2025-09-16
AI Technical Summary
Existing tetraphenylethylene derivatives have problems with selectivity and sensitivity when detecting cysteine, making it difficult to achieve efficient and sensitive detection and bioimaging applications.
A tetraphenylethylene derivative TPE-1s crystal probe was prepared. Through a specific chemical reaction synthesis method, a TPE-1s crystal probe with a monoclinic crystal structure was obtained. The substituents on the benzene ring were used to change the electron cloud distribution and fluorescence properties, thereby achieving highly selective recognition of cysteine and fluorescent signal indication.
It achieves highly selective recognition and highly sensitive detection of cysteine, with obvious changes in fluorescence signals, making it suitable for imaging in cells or organisms, and providing a tool for studying cysteine biological processes.
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Figure CN120647558A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the technical field of tetraphenylethylene derivatives, and particularly relates to a tetraphenylethylene derivative TPE-1s crystal probe and a preparation method and application thereof. Background Art
[0002] Tetraphenylethylene derivatives are a class of compounds derived from tetraphenylethylene by introducing various substituents onto its benzene rings. Tetraphenylethylene possesses a rigid, planar structure, with a central carbon atom connected to four benzene rings, forming a highly symmetrical spatial configuration. By introducing substituents such as hydroxyl, amino, carboxyl, and halogen atoms at various positions on the benzene rings, these derivatives can alter the electron cloud distribution, steric hindrance, and chemical activity of the molecules, thereby imparting them with diverse physicochemical properties and functions.
[0003] Tetraphenylethylene derivatives play an important role in the detection of cysteine (Cys), mainly in the following aspects:
[0004] (1) Highly selective recognition: Tetraphenylethylene derivatives can specifically bind to cysteine through specific chemical reactions or intermolecular interactions, thereby achieving selective recognition of cysteine.
[0005] (2) Fluorescence signal indication: Many tetraphenylethylene derivatives have aggregation-induced emission (AIE) properties. When detecting cysteine, their fluorescence properties will change significantly before and after binding to cysteine, such as increased fluorescence intensity or changed emission wavelength. This change in fluorescence signal can be used as an intuitive indicator for detecting cysteine. Accurate measurement using instruments such as fluorescence spectrometers can achieve high-sensitivity detection of cysteine.
[0006] (3) Bioimaging applications: Due to the good biocompatibility and fluorescence properties of tetraphenylethylene derivatives, they can be used to detect cysteine and perform imaging in cells or organisms. This allows for intuitive observation of the distribution and dynamic changes of cysteine in biological systems, providing a powerful tool for studying the mechanism of action of cysteine in biological processes and contributing to a deeper understanding of related physiological and pathological processes.
[0007] Therefore, it is of great significance to develop tetraphenylethylene derivatives and their preparation methods. Summary of the Invention
[0008] The present invention aims to provide a tetraphenylethylene derivative TPE-1s crystal probe, a preparation method thereof, and application thereof in detecting cysteine.
[0009] The implementation process of the present invention is as follows:
[0010] A tetraphenylethylene derivative TPE-1s crystal probe, the molecular formula of the crystal probe is C32 H 23 N3O6S, with a melting point of 113-115°C, belongs to the monoclinic system, with a space group of P 21 / n and unit cell parameters of α=90°, β=93.19°, γ=90°,
[0011] The preparation method of the tetraphenylethylene derivative TPE-1s crystal probe comprises the following steps:
[0012] (1) 1-(4-aminophenyl)-1,2,2-triphenylethylene and 2,4-dinitrobenzenesulfonyl chloride were used as raw materials, dichloromethane was used as solvent, cesium carbonate was added, and the reaction was carried out at room temperature. After filtration, purification, and drying, a yellow powder solid TPE-1 was obtained;
[0013] (2) The yellow powder solid TPE-1 was dissolved in methanol, filtered, and the clear liquid was allowed to stand and evaporate to obtain a dark yellow block-like tetraphenylethylene derivative TPE-1s crystal probe.
[0014] Furthermore, in step (1), the molar ratio of 1-(4-aminophenyl)-1,2,2-triphenylethylene, 2,4-dinitrobenzenesulfonyl chloride and cesium carbonate is 0.1:0.76:0.091.
[0015] Furthermore, in step (1), the reaction time at room temperature is 1 hour.
[0016] Furthermore, in step (1), the eluent of the purification process is a mixture of dichloromethane and petroleum ether in a volume ratio of 100:1; and a basic alumina column is used in the purification process.
[0017] Furthermore, in step (2), the clear liquid is allowed to stand for volatilization for 1 week.
[0018] Application of the above-mentioned tetraphenylethylene derivative TPE-1s crystal probe in the detection of cysteine.
[0019] Furthermore, the detection limit of the tetraphenylethylene derivative TPE-1s crystal probe for cysteine is 26.7 nmol·L -1 .
[0020] Positive effects of the present invention:
[0021] (1) The method of the present invention is simple and easy to operate.
[0022] (2) The present invention obtains two isomer crystals TPE-1s and TPE-1t by recrystallization. Among them, in the structure of TPE-1s, the angle between the fluorescent group TPE and the recognition group 2,4-dinitrobenzenesulfonyl structure is 80.399°, which is stretched. The benzene rings of two adjacent molecules are perpendicular to each other and parallel to each other, and there is a π-π stacking effect. The distance between the two adjacent benzene rings is and Furthermore, TPE-1s has high selectivity for Cys. BRIEF DESCRIPTION OF THE DRAWINGS
[0023] Figure 1 This is the synthesis route of the tetraphenylethylene derivative TPE-1s crystal probe of the present invention;
[0024] Figure 2 This is the H NMR spectrum of the yellow powder solid TPE-1 obtained in Example 1;
[0025] Figure 3 This is the NMR carbon spectrum of the yellow powder solid TPE-1 obtained in Example 1;
[0026] Figure 4 This is the mass spectrum of the yellow powder solid TPE-1 obtained in Example 1;
[0027] Figure 5 Actual images of TPE-1s crystal and TPE-1t crystal, where (a) TPE-1s crystal, (b) TPE-1t crystal;
[0028] Figure 6 Crystal structure diagrams of TPE-1s and TPE-1t, where (a) (c) TPE-1s crystal, (b) (d) TPE-1t crystal;
[0029] Figure 7 (a) Fluorescence emission spectra of TPE-1s and TPE-1t interacting with different substances; (b) Fluorescence intensity changes at 553 nm before and after the addition of Cys to a TPE-1s solution in EtOH / PBS (V:V = 7:3, pH = 7.4) in the presence of other substances.
[0030] Figure 8 Figure 2 is the fluorescence selectivity of TPE-1s and TPE-1t to Cys;
[0031] Figure 9 (a) Fluorescence spectra of probe TPE-1s to different concentrations of Cys (0-5.0 equiv.), (b) linear plot of fluorescence intensity of probe TPE-1s at 553 nm versus different Cys concentrations;
[0032] Figure 10The morphology changes before and after the action of TPE-1s and Cys, where (a, c) TPE-1s, (b, d) TPE-1s+Cys. DETAILED DESCRIPTION
[0033] The present invention will be further described below with reference to the embodiments.
[0034] In the first aspect, the present invention provides a tetraphenylethylene derivative TPE-1s crystal probe, the molecular formula of the crystal probe is C 32 H 23 N3O6S, with a melting point of 113-115°C, belongs to the monoclinic system, with a space group of P 21 / n and unit cell parameters of α=90°, β=93.19°, γ=90°,
[0035] In the second aspect, the present invention provides a method for preparing the above-mentioned tetraphenylethylene derivative TPE-1s crystal probe, the synthesis route is shown in Figure 1 , including the following steps:
[0036] (1) 1-(4-aminophenyl)-1,2,2-triphenylethylene and 2,4-dinitrobenzenesulfonyl chloride were used as raw materials, dichloromethane was used as solvent, cesium carbonate was added, and the reaction was carried out at room temperature. After filtration, purification, and drying, a yellow powder solid TPE-1 was obtained;
[0037] (2) The yellow powder solid TPE-1 was dissolved in methanol, filtered, and the clear liquid was allowed to stand and evaporate to obtain a dark yellow block-like tetraphenylethylene derivative TPE-1s crystal probe.
[0038] The 1-(4-aminophenyl)-1,2,2-triphenylethylene described in the present invention was prepared according to the McMurray reaction reported in the literature (Y. Cai, L. Li, Z. Wang, J. Z. Sun, A. Qin and B. Z. Tang, Chem. Commun., 2014, 50, 8892–8895). The synthesis method of the 1-(4-aminophenyl)-1,2,2-triphenylethylene is as follows: Under nitrogen protection, 0.985 g (15 mmol) of zinc powder and 20 mL of dry tetrahydrofuran (THF) were added to a three-necked flask. The suspension was cooled to -5 to 0°C, and then 0.83 mL of titanium tetrachloride (TiCl4, 7.5 mmol) was slowly added. The suspension was warmed to room temperature and stirred for 0.5 h, then heated to reflux for 2.5 h. The mixture was cooled to -5 to 0 ° C again, and a solution of 4-aminobenzophenone (0.495 g, 2.5 mmol) and benzophenone (0.455 g, 2.5 mmol) dissolved in 7.5 mL of tetrahydrofuran was added dropwise. After the addition was completed, the reaction mixture was heated to reflux and monitored by thin layer chromatography (TLC) until the carbonyl compound was completely consumed. The reaction was quenched with a 10% by mass aqueous solution of potassium carbonate (K2CO3) and extracted with ethyl acetate (EtOAc). The organic layer was collected and concentrated. The crude product was purified by column chromatography to obtain a white product 1-(4-aminophenyl)-1,2,2-triphenylethylene (TPE-NH2) with a yield of 38.0% and 0.33 g.
[0039] In a third aspect, the present invention provides the use of the tetraphenylethylene derivative TPE-1s crystal probe in detecting cysteine. The detection limit of the tetraphenylethylene derivative TPE-1s crystal probe for cysteine is 26.7 nmol·L -1 .
[0040] Example 1
[0041] A method for preparing a tetraphenylethylene derivative TPE-1s crystal probe comprises the following steps:
[0042] (1) 1-(4-Aminophenyl)-1,2,2-triphenylethylene (65.96 mg, 0.1 mmol) and 2,4-dinitrobenzenesulfonyl chloride (203.4 mg, 0.76 mmol) synthesized according to the literature were added to a 50 mL round-bottom flask and dissolved in 10 mL of dry dichloromethane. Cesium carbonate (30.1 mg, 0.091 mmol) was then added and stirred at room temperature for 1 hour. The solvent was removed under reduced pressure to obtain the crude product, which was eluted on a basic alumina column with dichloromethane and petroleum ether (volume ratio 100:1) to obtain TPE-1, a yellow powder solid, with a yield of 60%.
[0043] The reaction equation is as follows:
[0044]
[0045] (2) Dissolve the yellow powder solid TPE-1 (10 mg) in 5 mL of methanol, then filter and transfer the clear solution to a clean test tube and slowly let it stand for evaporation. After one week, a dark yellow block of tetraphenylethylene derivative TPE-1s crystal probe is obtained. Figure 5 a. TPE-1s crystal structure data were collected using a Bruker-AXS SMART APEX II instrument. Mo Kα diffraction (λ = 0.154184 nm) was collected at 298 K. The structure was refined using the SHELX-2014 / 7 program.
[0046] The yellow powder solid TPE-1 obtained in step (1) of the present invention has mp 113~115℃ and its H NMR spectrum is Figure 2 , 1 H NMR (DMSO-d6, 400MHz) δ: 10.92 (s, 1H), 8.92 (s, 1H), 8.43 (m, 7H), 8.40 (d, 6H), 8.10 (m, 6H), 6.90 (d, 2H).
[0047] Its NMR carbon spectrum is shown Figure 3 , 13 C NMR (100MHz, d6-DMSO) δ: 141.78, 139.78, 132.39, 136.65, 130.99, 128.46, 127.84, 128.41, 127.20, 123.06, 109.41.
[0048] Its mass spectrum is shown in Figure 4 , HR-ESI-MS calculation for C 32 H 23 N3O6SNa[M+Na] + ,600.1205,found600.1199.
[0049] The molecular formula of the TPE-1s crystal probe was C 32 H 23 N3O6S, with a melting point of 113-115°C, is dark yellow and blocky. It belongs to the monoclinic system, with a space group of P21 / n and unit cell parameters of α=90°, β=93.19°, γ=90°, In the TPE-1s structure, the angle between the fluorescent group TPE and the recognition group 2,4-dinitrobenzenesulfonyl is 80.399°, showing a stretched shape. The benzene rings of the two adjacent molecules are perpendicular and parallel to each other, and there is a π-π stacking effect. The distance between the two adjacent benzene rings is and Crystal structure see Figure 6 a. Figure 6 c shows the hydrogen bonding between TPE-1s molecules.
[0050] Comparative Example 1
[0051] The preparation method is the same as that described in Example 1, except that the solvent methanol is replaced by a mixed solvent of dichloromethane and petroleum ether with a volume ratio of 1:1, then filtered, the clear liquid is transferred to a clean test tube and slowly allowed to evaporate. After one week, orange needle-shaped crystals of tetraphenylethylene derivative TPE-1t are obtained, as shown. Figure 5 b. TPE-1t crystal structure data were collected using a Bruker-AXSSMART APEX II instrument, Mo Kα diffraction (λ = 0.154184 nm) was collected at 298 K, and the structure was refined using the SHELX-2014 / 7 program.
[0052] The molecular formula of the TPE-1t crystals was C 32 H 23 N3O6S, with a melting point of 113-115°C, is orange and needle-shaped, belongs to the triclinic system, has a space group of P-1, and a unit cell parameter of α=90°, β=94.37°, γ=90°, In the TPE-1t structure, the angle between the fluorescent group TPE and the recognition group 2,4-dinitrobenzenesulfonyl is 48.956°, which is more distorted than the spatial result of TPE-1s. The benzene rings of two adjacent molecules are only perpendicular to each other, and there is a π-π stacking effect. The distance between the two adjacent benzene rings is and The crystal structure is shown in Figure 6 b, Figure 6 d shows the hydrogen bonding between TPE-1t molecules.
[0053] Performance testing:
[0054] (1) Fluorescence selectivity of TPE-1s and TPE-1t crystals for ions or thiol small molecules
[0055] The fluorescence selectivity of TPE-1s crystal and TPE-1t crystal for Cys was studied by fluorescence competition experiment. + ,Na+ ,Mg 2+ ,F - ,Cl - ,Br - ,I - ,ClO4 - ,ClO - ,HSO3 - ,SO3 2- ,SO4 2- ,S2O3 2- ,CO3 2- ,HCO3 - ,SeO3 2- ) and thiol small molecules (Hcy, Cys, GSH, HS - ,S 2- ).
[0056] The fluorescence selectivity experimental process of TPE-1s crystals was as follows: 10 mL of 10 μmol·L -1 100 μL of 1 mmol·L TPE-1s crystals in EtOH / PBS (V:V=7:3, pH=7.4) was added. -1 (1 equivalent) different ions (K + ,Na + ,Mg 2+ ,F - ,Cl - ,Br - ,I - ,ClO4 - ,ClO - ,HSO3 - ,SO3 2- ,SO4 2- ,S2O3 2- ,CO3 2- ,HCO3 - ,SeO3 2- ) or thiol small molecules (Hcy, Cys, GSH, HS - ,S 2- ).
[0057] The fluorescence selectivity experimental process of the TPE-1t crystal obtained in Comparative Example 1 was as follows: 10 mL of 10 μmol·L -1 100 μL of 1 mmol·L TPE-1t crystals in EtOH / PBS (V:V=7:3, pH=7.4) was added. -1 (1 equivalent) different ions (K + ,Na + ,Mg 2+ ,F - ,Cl - ,Br- ,I - ,ClO4 - ,ClO - ,HSO3 - ,SO3 2- ,SO4 2- ,S2O3 2- ,CO3 2- ,HCO3 - ,SeO3 2- ) or thiol small molecules (Hcy, Cys, GSH, HS - ,S 2- ).
[0058] The results are as follows Figure 7 As shown in a, TPE-1s and TPE-1t themselves have weak fluorescence signals at 450nm. After adding Cys, the excitation wavelength is 395nm, and the probe TPE-1s shows a strong fluorescence emission peak at 553nm (yellow line). The fluorescence intensity is higher than that of TPE-1t+Cys (blue line). At the same time, the fluorescence intensity is also higher than that of TPE-1s or TPE-1t with the addition of any other ions or thiol small molecules (other colors). Under ultraviolet light, the solution changes from colorless to bright blue ( Figure 7 (a, inset). These results demonstrate that TPE-1s crystals are highly selective for Cys.
[0059] (2) Study on the interference of other ions or thiol small molecules on Cys detection
[0060] Interference experiment process of TPE-1s crystal: add 10mL 10μmol·L -1 100 μL of 1 mmol·L TPE-1s crystals in EtOH / PBS (V:V=7:3, pH=7.4) was added. -1 (1 equivalent) of Cys, and then 100 μL of 1 mmol·L -1 (1 equivalent) of other ions (K + ,Na + ,Mg 2+ ,F - ,Cl - ,Br - ,I - ,ClO4 - ,ClO - ,HSO3 - ,SO3 2- ,SO4 2- ,S2O3 2- ,CO3 2- ,HCO3 - ,SeO32- ) or small molecules (Hcy, GSH, HS - ,S 2- ), and measure its fluorescence emission spectrum, see Figure 7 b. Comparative analysis: TPE-1s plus other ions or small molecules, without Cys. The results show that the presence of other ions or small molecules has no significant effect on Cys detection by TPE-1s.
[0061] (3) Fluorescence selectivity of TPE-1s and TPE-1t crystals for Cys
[0062] The fluorescence selective detection ability of TPE-1s crystals and TPE-1t crystals for Cys in ethanol solution was studied through fluorescence emission experiments.
[0063] Experimental procedure: Add 10 mL of 10 μmol·L -1 Add 100 μL of 1 mmol·L TPE-1s crystal or TPE-1t crystal EtOH / PBS (V:V=7:3, pH=7.4) solution -1 (1 equivalent) of Cys, and measured its fluorescence emission spectrum.
[0064] from Figure 8 As can be seen, when one equivalent of Cys is added, TPE-1s exhibits a distinct fluorescence emission peak at 525 nm, with an intensity reaching 1000. In contrast, when one equivalent of Cys is added to TPE-1t, no significant fluorescence emission change is observed at 525 nm. This demonstrates the high fluorescence selectivity of TPE-1s crystals for Cys.
[0065] (4) Fluorescence titration experiment of TPE-1s crystals
[0066] In 10 mL 10 μmol·L -1 In TPE-1s EtOH / PBS (V:V=7:3, pH=7.4) solution, as the Cys concentration increases from 0 to 5 equivalents, the fluorescence intensity of the system increases continuously ( Figure 9 a), according to the fluorescence titration data, the linear relationship between Cys concentration change and fluorescence intensity at 553 nm was obtained ( Figure 9 b)y=11.955x-25.242,R 2 =0.9987, and the slope k is 11.995 μmol·L -1 The detection limit of TPE-1s crystal probe for Cys is 26.7 nmol·L -1 .
[0067] (5) Morphology before and after the interaction of TPE-1s with Cys
[0068] The morphological changes of the probe TPE-1s before and after the interaction with Cys were studied by scanning electron microscopy (SEM). TPE-1s alone aggregated into spherical structures in H2O / THF (9:1, v / v) ( Figure 10 a, c), add 100 μL 1 mmol·L -1 After Cys, the spherical structure disappears, a layered morphology is presented, and the aggregate size becomes smaller ( Figure 10 b,d), which is also the reason for the change in probe fluorescence intensity after the addition of Cys.
[0069] Those skilled in the art will readily conceive of other implementations of the present application after considering the specification and practicing the contents disclosed herein. This application is intended to cover any variations, uses, or adaptations of the present application that follow the general principles of the present application and include common knowledge or customary techniques in the art that are not disclosed in this application. The description and examples are to be considered merely as exemplary, and the present application is not limited to the precise structures described above and shown in the drawings, and various modifications and changes may be made without departing from the scope thereof.
Claims
1. A tetraphenylethylene derivative TPE-1s crystal probe, characterized by: The molecular formula of the crystal probe is C 32 H 23 N3O6S, with a melting point of 113-115°C, belongs to the monoclinic system, with a space group of P 21 / n and unit cell parameters of α=90°, β=93.19°, γ=90°, 2. The method for preparing the tetraphenylethylene derivative TPE-1s crystal probe according to claim 1, characterized in that: The steps include: (1) 1-(4-aminophenyl)-1,2,2-triphenylethylene and 2,4-dinitrobenzenesulfonyl chloride were used as raw materials, dichloromethane was used as solvent, cesium carbonate was added, and the reaction was carried out at room temperature. After filtration, purification, and drying, a yellow powder solid TPE-1 was obtained; (2) The yellow powder solid TPE-1 was dissolved in methanol, filtered, and the clear liquid was allowed to stand and evaporate to obtain a dark yellow block-like tetraphenylethylene derivative TPE-1s crystal probe.
3. The method for preparing the tetraphenylethylene derivative TPE-1s crystal probe according to claim 2, characterized in that: In step (1), the molar ratio of 1-(4-aminophenyl)-1,2,2-triphenylethylene, 2,4-dinitrobenzenesulfonyl chloride and cesium carbonate is 0.1:0.76:0.
091.
4. The method for preparing the tetraphenylethylene derivative TPE-1s crystal probe according to claim 2, characterized in that: In step (1), the reaction time at room temperature is 1 hour.
5. The method for preparing the tetraphenylethylene derivative TPE-1s crystal probe according to claim 2, characterized in that: In step (1), the eluent for the purification process is a mixture of dichloromethane and petroleum ether in a volume ratio of 100:1; and a basic alumina column is used in the purification process.
6. The method for preparing the tetraphenylethylene derivative TPE-1s crystal probe according to claim 2, characterized in that: In step (2), the clear solution is allowed to stand for 1 week for volatilization.
7. Use of the tetraphenylethylene derivative TPE-1s crystal probe according to claim 1 in detecting cysteine.
8. The use according to claim 7, characterized in that: The detection limit of the tetraphenylethylene derivative TPE-1s crystal probe for cysteine is 26.7 nmol·L -1 .