CN106632063B - Compound I and compound II based on phenanthroimidazole and its preparation method and application - Google Patents

Abstract

The present invention relates to compound I and compound II and their preparation methods and applications. By introducing different substituent structures at Ar, the adjustment of the luminous color and efficiency of the above compounds can be realized; the loss of efficiency of the compounds in the solid state can be effectively curbed, and the realization of Its high fluorescence quantum yield under water system conditions; by selecting different aminocarboxylic acid structures as reactants, introducing different carboxylic acid structures at the N1 benzene ring, and preparing various carboxylic acid derivative structures by appropriate methods to improve The molecular compatibility and recognition ability of the above compound structure. Compound I and compound II are a new type of probe structure, phenanthroimidazole carboxylic acid and carboxylic acid derivatives can freely enter and exit cells, and selectively recognize and image specific structures, making them useful in biology, medicine, health and The field of monitoring has extremely broad application prospects.

Description

Compound I and compound II based on phenanthroimidazole and its preparation method and application

technical field

The present invention relates to the technical field of analysis and detection, more specifically, relates to compound I and compound II and their preparation methods and applications.

Background technique

With the rapid development of the national economy and the continuous improvement of people's living standards, real-time monitoring of personal health, disease prevention and treatment have gradually become people's livelihood issues that people pay more attention to. Compared with the delay of traditional in vitro chemical testing and the harmfulness of online radiation detection, fluorescence online imaging technology has gradually entered people's field of vision with its advantages of high efficiency, greenness and strong real-time performance, and has been widely used in cellular immunology, microbiology, etc. Science, molecular biology, genetics, neurobiology, pathology, oncology, clinical laboratory science, medicine, botany and other fields of scientific research and people's livelihood.

The key technology of fluorescence imaging technology is the selection of fluorescent substances as labeled probes (or dyes). Ideal probe molecules are specifically adsorbed on specific cells and tissues through physical or chemical actions, and realize two-dimensional or three-dimensional imaging under low-energy optical irradiation, and judge cells or the health of the organization. Compared with ordinary chemical staining, the sensitivity of fluorescent staining is 100-1000 times higher, and online analysis of living organisms can be realized through appropriate functional modification.

Usually, the structure of a fluorescent probe consists of 2-3 parts, namely a luminescence motif, a link motif (optional) and a recognition motif. Among them, the luminescent element is mainly responsible for achieving high-efficiency luminescence under external environmental stimuli (light, electricity, or magnetic action), and then detected by external signal detection equipment and turned into visual information; the recognition element is mainly for targeting cells or the body. The target plays the role of identification to improve the positioning ability of the probe; the link primitive is mainly to link the two, so that the light-emitting position and the structure to be detected have a stable corresponding relationship, ensuring the accuracy of the detection process. In practical applications, in order to simplify the preparation process of the fluorescent probe structure and reduce the cost, the recognition motif and the luminescent core of the fluorescent probe can also be served by the same chemical structure in the probe molecule, but this has something to do with the luminescent mechanism and recognition of the compound. There is a great relationship, neither mutual interference, but also fast response and high resolution, this will be the mainstream direction of future development of fluorescent probes.

As a new type of luminescent material, phenanthrene imidazole has high luminous efficiency, excellent photothermal stability, relatively balanced carrier injection and transport capabilities, and has shown great application potential in the field of luminescent materials; at the same time, phenanthrene And imidazole is also the parent structure of a class of anticancer drugs, indicating that it has certain molecular recognition and localization capabilities. Although the combination of the above two advantages makes phenanthroimidazole meet the requirements of an ideal fluorescent probe building block, the biocompatibility of highly aromatic fused rings such as phenanthroimidazole is poor, and the ability to recognize and position is poor. Poor, it cannot be used as a fluorescent probe, and other groups need to be introduced to adjust its performance.

Contents of the invention

The object of the present invention is to provide the general structural formula as the compound of formula I and formula II and its preparation method and application, i.e. compound I and compound II and its preparation method and application, specifically to provide a class of carboxyl substituted based on phenanthroimidazole N1 The acid and carboxylic acid derivatives solve the problem that the phenanthroimidazole compounds in the prior art cannot be used as fluorescent probes.

The technical solution adopted by the present invention to solve the technical problem is: a compound of formula I has the following structural formula:

Among them, Ar represents an aromatic group or its derivative structure, and Ar is selected from phenyl, tolyl, naphthyl, furyl, thienyl, pyrrolyl, pyridyl, pyranyl, quinolinyl, indolyl, carbazole R group, anilino group, triphenylethylene, tetraphenylethylene and their corresponding derivative structures; R is selected from hydroxyl, halogen, alkoxy group, aromatic phenol group, amino group and aromatic amino group and its corresponding One of the derivatives; G1 and G2 respectively represent any substituent group at any position on the phenanthrene ring.

In the compounds of formula I according to the invention, G1 and G2 each represent hydrogen.

The present invention also provides a preparation method for the compound of the above formula I, comprising the steps of:

S1, the aromatic aldehyde derivative Phenanthrenequinone Derivatives Ammonium acetate and aminobenzoic acid Heating to 80°C-180°C in an organic solvent, cooling to room temperature after reacting for 1-24 hours, filtering to obtain

Wherein, Ar is selected from phenyl, tolyl, naphthyl, furyl, thienyl, pyrrolyl, pyridyl, pyranyl, quinolinyl, indolyl, carbazolyl, anilino, triphenylethylene, One of tetraphenylethylene and its corresponding derivative structures; G1 and G2 each represent any substituent group at any position on the phenanthrene ring.

Step S2 is also included after step S1:

S2, the prepared in the step S1 The hydroxyl groups in the group are subjected to corresponding substitution reactions to obtain the compound of formula I substituted by R, wherein R is selected from one of halogen, alkoxy group, aromatic phenol group, amino group, aromatic amino group and their corresponding derivatives. kind.

In the preparation method of the compound of formula I of the present invention, in step S1, preferably heating to 100°C-120°C; preferably cooling to room temperature after reacting for 2-4 hours; the organic solvent is preferably selected from acetic acid, tetrahydrofuran, toluene, Any one or more of benzene, N,N-dimethylformamide, N,N-dimethylacetamide and N-methylpyrrolidone, most preferably acetic acid and the mixed system of acetic acid and other solvents, namely acetic acid Mixed system with any one or more of tetrahydrofuran, toluene, benzene, N,N-dimethylformamide, N,N-dimethylacetamide and N-methylpyrrolidone.

Further, step S3 may also be included between step S1 and step S2: for the obtained For further purification, recrystallization and column chromatography can be used, wherein the recrystallization solvent is preferably tetrahydrofuran, ethanol, toluene or N,N-dimethylformamide, more preferably N,N-dimethylformamide.

The present invention also provides another compound of formula II, which has the following general structural formula:

Among them, Ar represents an aromatic group or its derivative structure, and Ar is selected from phenyl, tolyl, naphthyl, furyl, thienyl, pyrrolyl, pyridyl, pyranyl, quinolinyl, indolyl, carbazole _One of base, anilino, triphenylethylene, tetraphenylethylene and their corresponding derivative structures; R and R1 are each selected from hydroxyl, halogen, alkoxy, aromatic phenol group, amino and aromatic One of the amine group and its corresponding derivatives; G1 and G2 each represent any substituent group at any position on the phenanthrene ring.

In the compounds of formula II according to the invention, G1 and G2 each represent hydrogen.

The present invention also provides a preparation method for the compound of the above formula II, comprising the following steps: Sa, aromatic aldehyde derivative Phenanthrenequinone Derivatives Ammonium acetate and aminophthalic acid Heating to 80°C-180°C in an organic solvent, cooling to room temperature after reacting for 1-24 hours, filtering to obtain

Wherein, Ar is selected from phenyl, tolyl, naphthyl, furyl, thienyl, pyrrolyl, pyridyl, pyranyl, quinolinyl, indolyl, carbazolyl, anilino, triphenylethylene, One of tetraphenylethylene and its corresponding derivative structures; G1 and G2 each represent any substituent group at any position on the phenanthrene ring.

In the preparation method of the compound of formula II, step Sb is also included after step Sa:

Sb, prepared in the Sa step The two hydroxyl groups in each undergo corresponding substitution reactions respectively to obtain the compound of formula II substituted by R and R ₁ respectively, wherein R and R ₁ are each selected from halogen, alkoxyl group, aromatic phenol group, amino group and One of aromatic amine groups and their corresponding derivatives.

In the preparation method of the compound of formula II of the present invention, in step Sa, it is preferably heated to 100°C-120°C; it is preferably cooled to room temperature after reacting for 2-4 hours; the organic solvent is preferably selected from acetic acid, tetrahydrofuran, toluene, Any one or more of benzene, N,N-dimethylformamide, N,N-dimethylacetamide and N-methylpyrrolidone, most preferably acetic acid and the mixed system of acetic acid and other solvents, namely acetic acid Mixed system with any one or more of tetrahydrofuran, toluene, benzene, N,N-dimethylformamide, N,N-dimethylacetamide and N-methylpyrrolidone.

Further, step Sc may also be included between step Sa and step Sb: for the step Sa obtained For further purification, recrystallization and column chromatography can be used, wherein the recrystallization solvent is preferably tetrahydrofuran, ethanol, toluene or N,N-dimethylformamide, more preferably N,N-dimethylformamide.

The above-mentioned compounds of formula I and formula II are respectively used for the preparation of fluorescent probes, especially for the preparation of biological fluorescent probes.

Implement the compound of formula I of the present invention and formula II and its preparation method and application, have following beneficial effect: the compound of formula I of the present invention and formula II is by introducing different substituent structures at Ar place (C2 substituent position), Realize the adjustment of the luminescent color and efficiency of the above compounds; focus on the introduction of triphenylethylene or tetraphenylethylene units and their derivative structures to effectively curb the loss of the efficiency of the compound in the solid state and achieve high efficiency under water system conditions. Fluorescence quantum yield; by selecting different aminocarboxylic acid structures as reactants, introducing different carboxylic acid structures at the N1 benzene ring, and preparing various carboxylic acid derivative structures by appropriate methods, such as acyl halides, aromatic acid alkyls Esters, aryl aryl esters, alkyl aryl amide or aryl aryl amide, etc., to improve the molecular compatibility and recognition ability of the above compound structures; emphasizing that a class of water stable Acyl chloride structure, and proved by crystal structure that intramolecular charge separation may be the key factor to stabilize this type of structure; Formula I and Formula II compounds are a new type of probe structure, phenanthroimidazole carboxylic acid and carboxylic acid derivatives can freely In and out of cells, and selective recognition and imaging of specific structures, it has extremely broad application prospects in the fields of biology, medicine, health and monitoring.

Description of drawings

Fig. 1A is a graph showing the relative fluorescence intensity change curve of the fluorescence spectrum of TPE-PA under different ratios of tetrahydrofuran and water;

Figure 1B is a graph showing the relative fluorescence intensity change curve of the fluorescence spectrum of TPE-PAC under different ratios of tetrahydrofuran and water;

Fig. 2 is a curve diagram of the pH value of TPE-PAC in water as a function of time;

Figure 3A is a bright field photograph of TPE-PAC stained Hela cells under a fluorescence microscope;

Figure 3B is a fluorescent photo of TPE-PAC stained Hela cells under a fluorescent microscope;

Fig. 4 is a graph showing the change in fluorescence spectrum of TPE-P2A under different ratios of tetrahydrofuran and water;

Figure 5A is a bright field photograph of TPE-P2A stained Hela cells under a fluorescence microscope;

Fig. 5B is a fluorescent photo of Hela cells stained with TPE-P2A under a fluorescent microscope.

Detailed ways

Below in conjunction with accompanying drawing and embodiment, compound I and compound II of the present invention and preparation method thereof and application are further described:

A class of compounds provided by the present invention can be used as the compound of formula I and formula II of novel fluorescent probe material, i.e. compound I and compound II, this material is phenanthroimidazole carboxylic acid and carboxylic acid derivative structure, and its structure has following general structure Mode:

The above structure takes 9,10-phenanthroimidazole as the core structure, and Ar represents an aromatic group or its derivative structure, which are respectively selected from phenyl, tolyl, naphthyl, furyl, thienyl, pyrrolyl, pyridyl, pyran One of base, quinolinyl, indolyl, carbazolyl, anilino, triphenylethylene, tetraphenylethylene, etc. and the corresponding derivative structures. R and R1 respectively represent the carbonyl link substituent structure _of any substitution position on the N1 substituted benzene ring in imidazole, which are respectively selected from hydroxyl, halogen, alkoxy or aromatic phenol groups, amino groups and aromatic amino groups. Carboxylic acid group derivative structure. G1 and G2 represent any substituent groups at any position on the phenanthrene ring.

Specific synthesis process:

(1)

The derivative structures of aromatic aldehydes and phenanthrenequinones were obtained through various coupling, substitution and condensation reactions. The derivatives of aromatic aldehydes and phenanthrenequinones are all existing compounds, and their synthesis process is prior art, and the specific preparation process will not be repeated here.

(2)

The above-mentioned aromatic aldehyde derivatives, phenanthrenequinone derivatives, ammonium acetate and aminobenzoic acid (or aminophthalic acid) were prepared in a "one-pot method" in an appropriate solvent and temperature to prepare important carboxylic acid structures.

Concrete operation: heating aromatic aldehyde derivatives, phenanthrenequinone derivatives, ammonium acetate and aminobenzoic acid (or aminophthalic acid) to an appropriate temperature in an organic solvent, cooling to room temperature after a period of reaction, and filtering to obtain a carboxylic acid structure ( Further purification using recrystallization and column chromatography). Among them, the organic solvent is preferably acetic acid, tetrahydrofuran, toluene, benzene, N,N-dimethylformamide (DMF), N,N-dimethylacetamide (DMAC) and N-methylpyrrolidone (NMP), etc. or two or a mixed system of the three, most preferably acetic acid and its mixed system with other solvents; the temperature is preferably 80°C to 180°C, and the most preferred is 100°C to 120°C; the reaction time is preferably 1 hour to 24 hours, and the most preferred 2 hours to 4 hours; recrystallization solvent, preferably tetrahydrofuran, ethanol, toluene, DMF, etc., most preferably DMF.

(3)

The above-mentioned carboxylic acid structure adopts the corresponding method to obtain the substituted acyl halide, alkyl arylate, aryl aryl ester, alkyl aryl amide or aryl aryl amide of phenanthroimidazole N1 benzene ring respectively. It should be noted that the specific substitution reaction processes are all prior art, and will not be described in detail here.

Further, G1 and G2 respectively represent hydrogen.

Further, when Ar is any aryl structure, substituent R and R ₁ are hydroxyl, halogen, alkoxyl group or aromatic phenol group, amine group and aromatic amine group and derivatives thereof, its preferred compound structure is:

Wherein, X is a kind of in F, Cl, Br and I, and R ₂ -R ₅ are respectively hydrogen, alkyl, hydroxyl, alkoxy, nitro, cyano, amino, mercapto, halogen atom etc. (further wherein the number of carbon atoms in the alkyl or alkoxy group is 1 to 12), Ar' and Ar" are each selected from phenyl, tolyl, naphthyl, furyl, thienyl, pyrrolyl, pyridyl, pyridyl One of pyryl, quinolinyl, indolyl, carbazolyl, anilino, triphenylethylene, tetraphenylethylene, etc. and the corresponding derivative structures.

Further, Ar is limited to a benzene ring or a benzene ring derivative, a naphthalene ring or a naphthalene ring derivative, an anthracycline or an anthracycline derivative substituent, and its compound general formula and preferred compound structure are as follows, and use Represents the derivative structure of carboxylic acid groups constructed by hydroxyl, halogen, alkoxy or aromatic phenol groups, amine groups and aromatic amine groups on the N1 benzene ring:

Wherein, A ₁ -A ₉ are each hydrogen, alkyl, hydroxyl, alkoxy, nitro, cyano, amino, mercapto, halogen atom, phenyl, tolyl, naphthyl, furyl, thienyl, pyrrole One of base, pyridyl, pyranyl, quinolinyl, indolyl, carboxylic acid or carboxylic acid derivatives, carbazolyl or anilino and corresponding derivative structures (further, wherein alkyl or alkoxy The number of carbon atoms in the group is 1 to 12).

Further, Ar is limited to triphenylethylene and triphenylethylene derivatives, its general formula and preferred compound structure are as follows, and use Represents the derivative structure of carboxylic acid groups constructed by hydroxyl, halogen, alkoxy or aromatic phenol groups, amine groups and aromatic amine groups on the N1 benzene ring:

Among them, Ar"' is selected from phenyl, tolyl, naphthyl, furyl, thienyl, pyrrolyl, pyridyl, pyranyl, quinolinyl, indolyl, carbazolyl, anilino, benzothio Oxadiazole and the corresponding derivative structure or one of the new aromatic groups composed of any two; B ₁ -B ₁₅ are each hydrogen, alkyl, hydroxyl, alkoxy, nitro, cyano, amino, Mercapto, halogen atom, phenyl, tolyl, naphthyl, furyl, thienyl, pyrrolyl, pyridyl, pyranyl, quinolinyl, indolyl, carboxylic acid or carboxylic acid derivatives, carbazolyl or One of the anilino groups or one of the corresponding derivatives (further, the number of carbon atoms in the alkyl or alkoxy group is 1 to 12).

Further, Ar is limited to tetraphenylethylene and tetraphenylethylene derivatives, its general formula and preferred compound structure are as follows, and use Represents the derivative structure of carboxylic acid groups constructed by hydroxyl, halogen, alkoxy or aromatic phenol groups, amine groups and aromatic amine groups on the N1 benzene ring:

Among them, Ar"' is selected from phenyl, tolyl, naphthyl, furyl, thienyl, pyrrolyl, pyridyl, pyranyl, quinolinyl, indolyl, carbazolyl, anilino, benzothio Oxadiazole and the corresponding derivative structure or one of the new aromatic groups composed of any two; B ₁ -B ₁₅ are each hydrogen, alkyl, hydroxyl, alkoxy, nitro, cyano, amino, Mercapto, halogen atom, phenyl, tolyl, naphthyl, furyl, thienyl, pyrrolyl, pyridyl, pyranyl, quinolinyl, indolyl, carboxylic acid or carboxylic acid derivatives, carbazolyl or One of the anilino groups or one of the corresponding derivatives (further, the number of carbon atoms in the alkyl or alkoxy group is 1 to 12).

Further, Ar is furan, thiophene, pyrrole, pyridine, pyran, quinoline (containing isoquinoline), indole, carbazole, anilino, benzothiadiazole and their corresponding derivative structures, or any two One of the new aromatic groups composed of those whose compound general formula and preferred compound structure are as follows, and use Represents the derivative structure of carboxylic acid groups constructed by hydroxyl, halogen, alkoxy or aromatic phenol groups, amine groups and aromatic amine groups on the N1 benzene ring:

In all above-mentioned structural formulas, wherein the preferred structure of aryl and alkyl can be selected from one of the 29 shown in the following formula or a hydrogen atom:

Embodiment 1: the synthesis of phenyl-phenanthroimidazole-N1-carboxylic acid and derivatives

React an appropriate amount of benzaldehyde, phenanthrenequinone, p-aminobenzoic acid and excess ammonium acetate in glacial acetic acid at 120°C for 3 hours, cool to room temperature, and filter to obtain a carboxylic acid derivative. Dissolve it in a mixed solvent of N,N-dimethylformamide and thionyl chloride, react at 80°C for 4 hours, and then spin off unreacted thionyl chloride. Excessive aniline was added to reflux for 4 hours, and the structure of the target compound was obtained through chromatographic column separation. ¹ H NMR(DMSO,500MHz):δ:10.6(s,1H),8.96(d,1H),8.90(d,1H),8.70(d,1H),8.21(d,2H),7.85(d, 2H),7.79(t,1H),7.71(t,1H),7.60-7.56(m,5H),7.40-7.37(m,7H),7.11(d,1H).MALDI-TOF(m/z) :[M+]calcd.C ₃₄ H ₂₃ N ₃ O, 489.5659; found, 490.59.

Embodiment 2: Synthesis of 1-naphthyl-phenanthroimidazole-N1-carboxylic acid and derivatives

React an appropriate amount of 1-naphthaldehyde, phenanthrenequinone, p-aminobenzoic acid and excess ammonium acetate in glacial acetic acid at 120°C for 3 hours, cool to room temperature, and filter to obtain a carboxylic acid derivative. Dissolve it in a mixed solvent of N,N-dimethylformamide and thionyl chloride, react at 80°C for 4 hours, and then spin off unreacted thionyl chloride. Excessive aniline was added to reflux for 4 hours, and the structure of the target compound was obtained through chromatographic column separation. ¹ H NMR (500MHz, DMSO-d ₆ ) δ13.27(s, 1H), 9.02-8.97(m, 1H), 8.94(d, J=8.3Hz, 1H), 8.67(dd, J=7.9, 1.4 Hz,1H), 8.02-7.93(m,4H),7.91(dd,J＝8.2,1.4Hz,1H),7.83-7.65(m,5H),7.61(ddd,J＝8.4,7.0,1.4Hz, 1H),7.60-7.45(m,3H),7.41(ddd,J＝8.1,6.9,1.1Hz,1H),7.14(dd,J＝8.3,1.2Hz,1H).MALDI-TOF(m/z) : [M+]calcd.C ₃₈ H ₂₅ N ₃ O, 539.6246; found, 540.77.

Embodiment 3: Synthesis of 2-naphthyl-phenanthroimidazole-N1-carboxylic acid and derivatives

React an appropriate amount of 2-naphthaldehyde, phenanthrenequinone, p-aminobenzoic acid and excess ammonium acetate in glacial acetic acid at 120°C for 3 hours, cool to room temperature, and filter to obtain a carboxylic acid derivative. Dissolve it in a mixed solvent of N,N-dimethylformamide and thionyl chloride, react at 80°C for 4 hours, and then spin off unreacted thionyl chloride. Excess aniline was added to reflux for 4 hours, and the structure of the target compound was obtained through chromatographic column separation. ¹ HNMR (500MHz, DMSO-d ₆ )δ10.36(s, 1H), 8.98(dd, J＝28.1, 8.4Hz, 2H), 8.67(dd, J＝8.0, 1.4Hz, 1H), 8.04-7.90 (m,5H),7.83-7.69(m,7H),7.66-7.49(m,4H),7.43(ddd,J＝8.2,7.0,1.1Hz,1H),7.35(t,J＝7.9Hz,2H ), 7.18 (dd, J=8.4, 1.2Hz, 1H), 7.11 (dd, J=8.0, 6.7Hz, 1H). MALDI-TOF (m/z): [M+] calcd. C ₃₈ H ₂₅ N ₃ O, 539.6246; found, 540.45.

Embodiment 4: Synthesis of tetraphenylethylene-phenanthroimidazole-N1-carboxylic acid and derivatives

Appropriate amount of tetrastyryl formaldehyde, phenanthrenequinone, p-aminobenzoic acid and excess ammonium acetate were reacted in glacial acetic acid at 120°C for 3 hours, cooled to room temperature, filtered, and column chromatographed to obtain the carboxylic acid derivative TPE-PA. MALDI-TOF (m/z): [M+] calcd. C ₄₈ H ₃₂ N ₂ O ₂ , 668.7799; found, 668.8732. Anal Calc. for C ₄₈ H ₃₂ N ₂ O ₂ : C, 86.20; H, 4.82; N, 4.19; O, 4.78. Found: C, 86.12; H, 4.77; N, 4.20; O, 4.74.

Dissolve TPE-PA in a mixed solvent of N,N-dimethylformamide (a small amount) and thionyl chloride, react at 80°C for 4 hours, spin off unreacted thionyl chloride, extract with water and dichloromethane After drying, chromatographic column separation to obtain the acid chloride compound structure TPE-PAC. MALDI-TOF (m/z): [M+]calcd. C ₄₈ H ₃₁ ClN ₂ O, 687.2255; found, 687.2345. AnalCalc. for C ₄₈ H ₃₁ ClN ₂ O: C, 83.89; H, 4.55; ; N, 4.08; O, 2.33 Found: C, 83.87; H, 4.53; N, 4.09; O, 2.32.

Example 5: Research on Aggregation-Induced Emission (AIE) Properties of TPE-PA and TPE-PAC

As shown in Figures 1A and 1B, they are the relative fluorescence intensity curves of the fluorescence spectra of TPE-PA and TPE-PAC under different ratios of tetrahydrofuran and water. As the water content increases, the fluorescence intensity of the two increases significantly, with The obvious aggregation-induced luminescent phenomenon is a kind of potential fluorescent molecular probe.

Embodiment 6: Study on the stability of TPE-PAC water system

As shown in Figure 2, it is the curve of the pH value of TPE-PAC in water versus time. Under normal conditions, acid chlorides will be rapidly hydrolyzed into corresponding carboxylic acids and hydrogen chloride in water, thereby losing high reactivity, but TPE-PAC can show remarkable stability in water system conditions due to the particularity of imidazole. Under the condition of stirring, its decomposition rate is estimated to be only 25% according to the change of pH value within 1500 minutes, and it is a kind of stable acid chloride structure in water system. Moreover, the acid chloride structure can be normally subjected to extraction and column chromatography, indicating that it can be used as a fluorescent molecular probe motif under aqueous conditions.

Embodiment 7: TPE-PAC cell staining experiment

After making TPE-PAC into a certain concentration of DMSO solution, drop it into the cell culture medium, select Hela cells as the research object, and observe its polarity with a fluorescence microscope after culturing for a period of time, as shown in Figure 3A and 3B, it was found that TPE -PAC can pass through the cell wall smoothly, and is enriched in the cytoplasm in a directional manner, thereby showing blue fluorescence, indicating that this type of acid chloride structure can be used as a fluorescent probe under aqueous conditions.

Example 8: Synthesis of tetraphenylethylene-phenanthroimidazole-N1-carboxylic acid derivative TPE-P2A

Appropriate amount of tetrastyryl formaldehyde, phenanthrenequinone, p-aminophthalic acid and excess ammonium acetate were reacted in glacial acetic acid at 120°C for 3 hours, cooled to room temperature, filtered, and column chromatographed to obtain the carboxylic acid derivative TPE-P2A. MALDI-TOF (m/z): [M+] calcd. C ₄₉ H ₃₂ N ₂ O ₄ , 712.7894; found, 712.7902. Anal Calc. for C ₄₉ H ₃₂ N ₂ O ₄ : C, 82.57; H, 4.53; N, 3.93; O, 8.98. Found: C, 82.48; H, 4.45; N, 3.9387; O, 8.68.

Example 9: Research on Aggregation-Induced Emission (AIE) Properties of TPE-P2A

As shown in Figure 4, it is the fluorescence spectrum change curve of TPE-P2A under different ratios of tetrahydrofuran and water. As the water content increases, the fluorescence intensity of the two increases significantly, and there is obvious aggregation-induced luminescence phenomenon, which can be used as a Fluorescent molecular probes.

Embodiment 10: TPE-P2A cell staining experiment

After making TPE-P2A into a certain concentration of DMSO solution, drop it into the cell culture medium, select Hela cells as the research object, and observe its polarity with a fluorescence microscope after culturing for a period of time, as shown in Figures 5A and 5B, it was found that TPE -P2A can pass through the cell wall smoothly and be enriched in the cytoplasm in a directional manner, thereby showing blue fluorescence, indicating that the dicarboxylic acid structure has self-activation properties and can be used as a fluorescent probe.

It should be understood that those skilled in the art can make improvements or changes based on the above description, and all these improvements or changes should fall within the protection scope of the appended claims of the present invention.

Claims (8)

1. a kind of compound of Formulas I, has the following structure general formula:

Wherein, Ar is selected from naphthalene, triphenylethylene or tetraphenylethylene；R is selected fromOr one of halogen；G1 and G2 respectively indicates hydrogen.

2. a kind of preparation method of the compound of Formulas I, which comprises the steps of:

S1, by fragrant aldehyde derivativesPhenanthrenequione derivativeAmmonium acetate and aminobenzoic acid? 80 DEG C -180 DEG C are heated in organic solvent, reaction is cooled to room temperature after 1-24 hours, is filtered, is obtained

Wherein, Ar is selected from naphthalene, triphenylethylene or tetraphenylethylene；G1 and G2 respectively indicate hydrogen.

3. the preparation method of compound according to claim 2, which is characterized in that further include step S2 after step S1:

S2, by what is be prepared in S1 stepIn hydroxyl carry out corresponding substitution reaction respectively and obtain By the compound of the R Formulas I replaced, wherein R is selected fromOr one of halogen.

4. the preparation method of compound according to claim 2, which is characterized in that in step sl, be heated to 100 DEG C- 120℃；Reaction is cooled to room temperature after 2-4 hours；The organic solvent is selected from acetic acid, tetrahydrofuran, toluene, benzene, N, N- diformazan Any one or more in base formamide, DMAC N,N' dimethyl acetamide and N-Methyl pyrrolidone.

5. a kind of compound of Formula II, has the following structure general formula:

Wherein, Ar indicates naphthalene, triphenylethylene or tetraphenylethylene；R and R₁Respectively it is selected fromOr in halogen One kind；G1 and G2 respectively indicate hydrogen.

6. a kind of preparation method of the compound of Formula II, which comprises the steps of:

Sa, by fragrant aldehyde derivativesPhenanthrenequione derivativeAmmonium acetate and amino phthalic acidIt is heated to 80 DEG C -180 DEG C in organic solvent, reaction is cooled to room temperature after 1-24 hours, is filtered, is obtained

Wherein, Ar is selected from naphthalene, triphenylethylene or tetraphenylethylene；G1 and G2 respectively indicate hydrogen.

7. the preparation method of compound according to claim 6, which is characterized in that further include step Sb after step Sa:

Sb, by what is be prepared in Sa stepIn two hydroxyls respectively carry out it is corresponding Substitution reaction obtains respectively by R and R₁The compound of substituted Formula II, wherein R and R₁Respectively it is selected fromOr halogen One of.

8. the purposes that compound described in claim 1 or 5 is used to prepare fluorescence probe.