CN108191902A - Glimmering analog derivative of a kind of 3,5- bisbenzimidazoles base -8- thienyl fluorine boron and its preparation method and application - Google Patents

Abstract

The invention provides a 3,5-dibenzimidazolyl-8-thienyl fluorine fluorine derivative, which belongs to the technical field of fluorine fluorine derivatives. The borofluorine derivatives have the structure shown in the following formula I: The invention also provides a preparation method of flubororfluorine derivatives. The synthesis route used in the present invention is C-H/N-H direct oxidative coupling reaction, compared with the traditional preparation technology of fluoroboron-fluoresceinization, it shortens the organic synthesis steps, avoids the cumbersome process of substrate pre-activation, and improves the The compatibility of the reaction increases the overall yield of the synthesis reaction. By applying the fluoroboron-fluorescein derivatives of the present invention to the specific fluorescence imaging and fluorescent labeling of the endoplasmic reticulum, the cost is much lower than that of commercially available endoplasmic reticulum labeling reagents, which can solve the problem of endoplasmic reticulum targeting in the prior art. The reagents have complex structures, inconvenient synthesis and high prices.

Description

A kind of 3,5-dibenzimidazolyl-8-thienylfluoroborofluorine derivatives and its preparation Methods and Applications

technical field

The invention belongs to the technical field of fluoborofluores derivatives, and specifically relates to a 3,5-dibenzimidazolyl-8-thienyl fluoborofluores derivatives and a preparation method and application thereof.

Background technique

The endoplasmic reticulum is a very important multifunctional organelle in eukaryotic cells. The synthesis, folding and modification of intracellular proteins, the synthesis of lipids and sterols take place in the endoplasmic reticulum. While completing these basic physiological functions, the endoplasmic reticulum has become a hub platform for coordinating signal transduction by virtue of its huge membrane structure, maintaining the stability of the intracellular environment. When genetic or environmental damage causes endoplasmic reticulum stress, excessive endoplasmic reticulum stress or prolonged endoplasmic reticulum stress can cause apoptosis, which can lead to a series of diseases [see: J.E.Vance, Traffic, 2015, 16, 1. ], specifically tracking and detecting the morphology and distribution of endoplasmic reticulum in cells is conducive to in-depth understanding and research on many related physiological activities [see: Z.Yang, Y.He, J.H.Lee, W.-S.Chae, W.X.Ren , J.H. Lee, C. Kang and J.S. Kim, Chem. Commun., 2014, 50, 11672.].

In recent years, there have been some reports on targeting reagents for labeling the endoplasmic reticulum, but the mechanism of the specific targeting and localization of the labeling is not very clear. We need more research to design reliable ER-labeled targeting reagents. Currently, combining targeting groups with fluorophores is commonly used to construct novel ER-targeting reagents [see: W.Xu,Z . Zeng, J.-H. Jiang, Y.-T. Chang and L. Yuan, Angew. Chem., Int. Ed. 2016, 55, 13658].

Among many fluorescent dyes, the dipyrromethane fluorescent dyes complexed with fluoroboron, also known as fluoroboronfluorescein, have a high molar extinction coefficient, high fluorescence quantum yield, stable spectral properties, high photothermal and chemical stability, With the advantages of small molecular weight and low cytotoxicity, it has been widely used as fluorescent probes such as biomolecules and ions and fluorescent reagents for organelle imaging [see: (a) S.Arai, S.-C.Lee, D.Zhai , M.Suzuki and Chang, YTSci.Rep.2014, 4, 6701; (b) X.Kong, F.Su, L.Zhang, J.Yaron, F.Lee, Z.Shi, Y.Tian and Meldrum, DR Angew. Chem., Int. Ed. 2015, 54, 12053; (c) L. Yang, Y.-J. Ji, J.-F. Yin, Y. Wu, H. Fan, Y. Zhang and G. -C. Kuang, Soft Matter, 2016, 12, 8581.]. There are many labeling molecules based on the fluoroborate skeleton that have been widely circulated in the market as standard labeling reagents for organelles, such as LysoTracker ^TM Red, LysoTracker ^TM Green, ER-Tracker ^TM Red and ER-Tracker ^TM Green [see: I. Johnson, MTZ Spence, The Molecular Probes Handbook, A Guide to Fluorescent Probes and Labeling Technologies, 11th Ed.; Molecular Probes: Eugene, OR, 2010.]. However, these common commercially available endoplasmic reticulum targeting reagents are complex in structure, inconvenient to synthesize, and expensive.

Therefore, providing a fluorescent molecule for endoplasmic reticulum labeling, which has a simple structure, easy synthesis, and low production cost has become an urgent problem to be solved by those skilled in the art.

Contents of the invention

The purpose of the present invention is to provide a small molecule of fluoroboron derivatives that can be used for endoplasmic reticulum labeling. Using the fluoroboron derivatives of the present invention for fluorescent labeling of endoplasmic reticulum can effectively solve the problem of endoplasmic reticulum. Plasma reticulum targeting reagents have complex structures, inconvenient synthesis, and high prices.

The invention also provides the preparation method and application of the fluorine boron derivatives.

The object of the invention is achieved through the following technical solutions:

A 3,5-dibenzimidazolyl-8-thienylfluoroboron derivative, which has the structure shown in the following formula I:

Wherein: R ₁ , R ₂ , R ₃ are one or two of hydrogen, deuterium, alkyl, alkoxy, carbonyl, ester, halogen, substituted aryl or substituted heteroaryl. Further, the carbon chain of the alkyl group, alkoxy group, ester group or carbonyl group is a straight chain, branched chain or cyclic chain with 0-40 carbons.

Further, the substituents in the substituted aryl and substituted heteroaryl are one or more of alkyl, alkoxy or carbonyl. Furthermore, the carbon chain of the alkyl group, alkoxy group or carbonyl group is a straight chain, branched chain or cyclic chain with 0-40 carbons.

As a specific example of a 3,5-dibenzimidazolyl-8-thienylfluoroboron derivative in the present invention, it has the structure shown in the following formula II:

A preparation method of 3,5-dibenzimidazolyl-8-thienylfluoroboron derivatives, comprising:

1) Mix 8-thienylfluoroborate with benzimidazole derivatives, oxidant and solvent evenly, and then react under anhydrous and oxygen-free conditions. The reaction formula is as follows:

2) After the reaction is completed, cool to room temperature, remove the solvent and add dichloromethane to dissolve the reaction system, then filter through diatomaceous earth, and wash with dichloromethane, combine the filtrates, remove the solvent under reduced pressure, and use a silica gel column for the residue The compound of formula I can be obtained by separation and purification by chromatography and vacuum drying.

Further, in the above step 1), the anhydrous and oxygen-free conditions can be protected by an inert gas, and further preferably, the reaction can be performed under a nitrogen atmosphere.

The synthetic route used in the preparation method of the present invention is C-H/N-H direct oxidative coupling reaction, compared with the traditional preparation technology of fluoborofluoresceinization, it shortens the comparatively tedious organic synthesis steps and avoids the problem of substrate pre-activation The cumbersome process improves the compatibility of the reaction and increases the total yield of the synthesis reaction.

As a specific example of the preparation method of a 3,5-dibenzimidazolyl-8-thienylfluoroboron derivative described in this invention, the oxidizing agent is copper acetate monohydrate, copper acetate, chlorine Copper chloride, copper bromide, ketone trifluoroacetate, copper (II) trifluoromethanesulfonate, copper acetylacetonate, silver carbonate, silver oxide, silver acetate, silver nitrate, silver hexafluoroantimonate, oxygen, iodobenzene acetate, One or more of benzoquinone, dichlorodicyanoquinone, sodium peroxodisulfate, ammonium peroxodisulfate, potassium peroxodisulfate, di-tert-butyl peroxide.

As a specific example of the preparation method of a kind of 3,5-dibenzimidazolyl-8-thienylfluoroborofluoro derivatives described in this invention, the solvent is methanol, ethanol, tetrahydrofuran, dichloromethane , chloroform, ether, dimethyl sulfoxide, benzene, o-dichlorobenzene, chlorobenzene, toluene, xylene, mesitylene, cyclohexane, petroleum ether, tert-amyl alcohol, 1,4-dioxane One or more of cyclic, 1,2-dichloroethane, N,N-dimethylformamide, N,N-dimethylacetamide.

As a specific example of the preparation method of a kind of 3,5-dibenzimidazolyl-8-thienyl fluorine fluorine derivatives described in this invention, the reaction concentration of the 8-thienyl fluorine fluorine is: 0.0001～10mol/L. More preferably, it is 0.1-8 mol/L, 0.5-5 mol/L, 1-3 mol/L.

As a specific example of the preparation method of a kind of 3,5-dibenzimidazolyl-8-thienylfluoroborofluoro derivatives described in this invention, the 8-thienylfluoroborofluoro, benzimidazole For derivatives, the molar ratio of the oxidizing agent is 1:(0.01～50):(0.01～100). More preferably, it is 1:(0.1-40):(10-80), 1:(5-30):(20-60).

As a specific example of the preparation method of a 3,5-dibenzimidazolyl-8-thienylfluoroborofluorine derivative described in this invention, the reaction temperature in step 1) is -40 to 160 °C, more preferably 20-100 °C, 40-80 °C; time is 0.1-720 h, more preferably 10-30 h.

The present invention also provides the application of the 3,5-dibenzimidazolyl-8-thienylfluoroborofluorine derivative, which is used in specific fluorescence imaging and fluorescent labeling of endoplasmic reticulum.

As a specific embodiment of the application of a 3,5-dibenzimidazolyl-8-thienylfluoroborophane derivative described in the present invention, the following steps are included:

Step 1: culturing cells in culture medium;

Step 2: removing the culture medium from the cultured cells, adding a buffer solution of 3,5-dibenzimidazolyl-8-thienylfluoroborophane derivatives, and culturing;

Step 3: After the culture in step 2 is completed, image the cultured glass-bottom dish through a fluorescent confocal microscope.

As a specific example of the application of a 3,5-dibenzimidazolyl-8-thienylfluoroborophane derivative in the present invention, the cells are HepG2 cells.

As a specific example of the application of a kind of 3,5-dibenzimidazolyl-8-thienyl fluoresbene derivatives of the present invention, the culture medium is DMEM (H ) culture medium.

As a specific example of the application of a 3,5-dibenzimidazolyl-8-thienylfluoroborofluorine derivative of the present invention, the 3,5-dibenzimidazolyl-8-thiophene The concentration of the buffer solution for the fluorescein-based derivatives is 2.5 μM in phosphate buffered saline.

As a specific example of the application of a 3,5-dibenzimidazolyl-8-thienylfluoroborophane derivative in the present invention, the culture condition in step 1 is 24 hours at 37°C.

As a specific embodiment of the application of a 3,5-dibenzimidazolyl-8-thienylfluoroborophane derivative described in the present invention, the following steps are included:

Add 5% CO ₂ to the DMEM (H) medium containing 10% fetal bovine serum, culture HepG2 cells at 37°C for 24 hours; remove the medium, add 2.5 μM compound 3,5-dibenzimidazole The phosphate buffer solution of 8-thienylfluoroborate, incubate for 30 minutes; after the incubation, take out the culture glass-bottom dish, wash it with phosphate buffer for 2 to 3 times, and then pass the culture glass-bottom dish through fluorescence coagulation. Focusing microscope imaging.

Compared with the prior art, the present invention has the following beneficial effects:

Compared with the existing commercially available fluorescent labeling reagents for endoplasmic reticulum, the synthetic route of 3,5-dibenzimidazolyl-8-thienylfluoroborofluorine derivatives of the present invention is more concise, efficient, environment-friendly, and the product price is Inexpensive and easily available in large quantities. The specific performance is:

1. The synthetic route used in the present invention is C-H/N-H direct oxidative coupling reaction. Compared with the traditional preparation technology of fluoroborate fluoresceinization, it shortens the comparatively tedious organic synthesis steps and avoids the problem of substrate preactivation. The cumbersome process improves the compatibility of the reaction and increases the total yield of the synthesis reaction.

2. Compared with the commercially available endoplasmic reticulum labeling reagents ER-Tracker Red and ER-Tracker Green, the compound of the present invention is simple to synthesize and inexpensive. The price of ER-Tracker Red and ER-Tracker Green is as high as 4863 yuan/100μg (Thermo Fisher Scientific Company), while our products are relatively cheap. Among the synthetic raw materials, even the most expensive AgOAc for oxidants has a cost price of Only 289 yuan/25g (Annaiji Company), and other synthetic raw materials are cheap and easy-to-obtain compounds on the market. The price of fluorine boron derivatives in this application is about 3000-8000 yuan/1g. At the same time, under the reaction conditions of the present invention, the yield can reach more than 20%. Therefore, the cost of using the fluoroborate derivatives of the present invention as endoplasmic reticulum fluorescent markers will be much lower than that of commercially available labeling reagents.

Description of drawings

Fig. 1 is the general structural formula of 3,5-dibenzimidazolyl-8-thienylfluoroboron derivatives.

Fig. 2 is the NMR spectrum H spectrum of 3,5-dibenzimidazolyl-8-thienylfluoroboronfluoride.

Fig. 3 is the NMR spectrum C of 3,5-dibenzimidazolyl-8-thienylfluoroboronfluoride.

Fig. 4 is the ultraviolet-visible light absorption spectrum and the fluorescence emission spectrum of 3,5-dibenzimidazolyl-8-thienylfluoroborate.

Fig. 5 is a cell imaging diagram of endoplasmic reticulum labeled with 3,5-dibenzimidazolyl-8-thienylfluoroborate.

Fig. 6 is the result of the CCK8 cytotoxicity experiment of 3,5-dibenzimidazolyl-8-thienylfluoroborate in HepG2 cells.

Detailed ways

In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

In the embodiment of the present invention, HepG2 cell line was purchased from ATCC (American Type Culture Collection), 10% fetal bovine serum was purchased from Hyclone Company, and DMEM (H) (Dulbecco's modifiedessential medium) medium was purchased from Gibco, USA. The endoplasmic reticulum dye ER-Tracker ^TM Green was purchased from Thermo Fisher Scientific.

Example 1

Synthesis of 3,5-Dibenzimidazolyl-8-thienylfluoroborofluorene

Add 8-thienylfluoroborate (13.7mg, 0.05mmol), benzimidazole (23.6mg, 0.20mmol), AgOAc (33.4mg, 4.0equiv), dimethyl sulfoxide (1.0mL) into the reaction tube, and Stir evenly under nitrogen, heat to 80°C, and react for 12 hours; the specific reaction formula is as follows:

After the reaction is complete, cool the reaction tube to room temperature, remove the solvent and add 10 mL of dichloromethane to dissolve the reaction system, then filter through diatomaceous earth and wash with 10-20 mL of dichloromethane, combine the filtrates, remove the solvent under reduced pressure, The residue was separated and purified by silica gel column chromatography (dichloromethane/petroleum ether/ethyl acetate=10:10:1, v/v/v), and after vacuum drying, the target product 3,5-dibenzimidazole was obtained as a black solid 5.1 mg of yl-8-thienylfluoroborophore, yield 20%.

The H-spectrum and C-spectrum of the product dibenzimidazolyl-8-thienylfluoroboronfluorescein in this example are shown in Figure 2 and Figure 3, respectively, and the structural data are characterized as follows:

¹ H NMR (400MHz, DMSO-d ₆ ) δ=7.13(d, J=4.4Hz, 2H), 7.33-7.39(m, 4H), 7.50-7.53(m, 3H), 7.68(d, J=2.0 Hz, 2H), 7.76-7.79(m, 2H), 7.96(s, 1H), 8.30(d, J=4.4Hz, 1H), 8.42(s, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ=111.4, 115.9, 120.1, 123.5, 124.4, 129.5, 131.7, 132.6, 133.2, 133.8, 134.8, 135.4, 138.6, 142.9, 143.7, 145.8. HRMS (ESI ⁺ ): calculated value C ₂₇ H ₁₈ BF ₂ N ₆ S[M+H] ⁺ : 507.1369, found value 507.1367.

Example 2

The ultraviolet-visible-near-infrared absorption spectrum and the fluorescence emission spectrum of the compound 3,5-dibenzimidazolyl-8-thienylfluoroborate prepared in Example 1

Dissolve the compound 3,5-dibenzimidazolyl-8-thienylfluoroborin in dichloromethane to make 1×10 ^-5 mol/L, put 2.5mL into a cuvette, measure the UV- Visible-near-infrared absorption and fluorescence emission spectra.

Figure 4 shows the UV-visible absorption spectrum and fluorescence emission spectrum of the compound 3,5-dibenzimidazolyl-8-thienylfluoroborate, wherein the black solid line represents the UV-visible absorption spectrum, and the black dotted line represents the fluorescence emission spectrum. As can be seen from Figure 4, the maximum absorption peak of the absorption spectrum of the compound 3,5-dibenzimidazolyl-8-thienylfluoroboron is located at 559nm; the maximum absorption peak of the fluorescence emission spectrum is located at 590nm, and the Stokes shift is 31nm.

Example 3

Fluorescent confocal confocal imaging of the compound 3,5-dibenzimidazolyl-8-thienylfluoroborin prepared in Example 1 and the commercially available endoplasmic reticulum stain ER-Tracker ^TM Green in HepG2 cells

First, 5% CO ₂ was introduced into the DMEM (H) medium containing 10% fetal bovine serum, and HepG2 cells were cultured at 37°C for 24 hours. Remove the culture medium, add 2.5 μM compound 3,5-dibenzimidazolyl-8-thienylfluoroborin in phosphate buffer, then add 1 μM commercially available endoplasmic reticulum stain ER-Tracker ^TM Green at 37°C co-incubation for 30 minutes. After the culture is over, take out the culture glass-bottom dish, wash it with phosphate buffer saline for 2 to 3 times, and image the culture glass-bottom dish through a fluorescent confocal microscope.

Fig. 5 is a fluorescence imaging diagram of the compound 3,5-dibenzimidazolyl-8-thienylfluoroboron (excitation wavelength: 553nm, emission wavelength collection range: 550-650nm). It ^can be seen visually that the compound 3,5-dibenzimidazole The distribution of yl-8-thienylfluoroborate in cells is basically the same as that of the commercially available endoplasmic reticulum stain ER-Tracker ^TM Green, and the corresponding Pearson's coefficient reaches 0.96, indicating that the compound 3,5-dibenzimidazolyl-8- Thienyl fluoroborate has an excellent endoplasmic reticulum tracer effect, and can specifically mark the endoplasmic reticulum in cells.

Example 4

CCK8 cytotoxicity test of the compound 3,5-dibenzimidazolyl-8-thienylflubororin prepared in Example 1

HepG2 cells in the logarithmic growth phase were inoculated into 96-well culture plates, with 3000 cells per well, at 37°C in DMEM (H) medium containing 10% fetal calf serum with 5% _CO2 Incubate overnight. After the cells were completely adhered to the wall, different concentrations of the compound 3,5-dibenzimidazolyl-8-thienylfluoroborate were added thereto, and three replicate wells and blank control wells were set up for each concentration group. The cells were cultured for 24 hours after adding the sample, and the cell viability was detected by the CCK8 assay.

Fig. 6 is the result of the CCK8 cytotoxicity experiment of 3,5-dibenzimidazolyl-8-thienylfluoroborate in HepG2 cells. As shown in Figure 6, in the concentration range of 0.25-4 μM, the cell survival rate of the compound 3,5-dibenzimidazolyl-8-thienylflubororine was very high (the survival rate was over 90%), and the cell survival rate at 8 μM Only when the survival rate is significantly reduced (survival rate is less than 30%), it shows that the toxicity of the compound 3,5-dibenzimidazolyl-8-thienylfluoroborate is very small and can be ignored in our labeling working concentration .

Example 5

In the preparation method of Example 1, hydrogen in R ₁ , R ₂ and R ₃ is replaced by deuterium, alkyl, alkoxy, carbonyl, ester group, halogen, substituted aryl or substituted heteroaryl, respectively. Other conditions remain unchanged, and a series of 3,5-dibenzimidazolyl-8-thienylfluoroborofluorine derivatives capable of realizing specific fluorescence imaging and fluorescent labeling of endoplasmic reticulum were successfully prepared. Among them, the product prepared in Example 1 has the best effect in fluorescence imaging and fluorescent labeling of endoplasmic reticulum.

Example 6

In the preparation method of embodiment 1 or 5, adjust relevant parameter and carry out series experiment:

In step 1), respectively select -40°C, -20°C, 0°C, 20°C, 40°C, 60°C, 100°C, 120°C, 140°C, 160°C instead of 80°C; the reaction time is controlled at 0.1-720h;

Respectively control 8-thienylfluoroboron derivatives and benzimidazole derivatives, the molar ratio of the oxidizing agent is 1:(0.01～50):(0.01～100);

Copper acetate monohydrate, copper acetate, copper chloride, copper bromide, ketone trifluoroacetate, copper (II) trifluoromethanesulfonate, copper acetylacetonate, silver carbonate, silver oxide, silver nitrate, silver hexafluoroantimonate , oxygen, iodobenzene acetate, benzoquinone, dichlorodicyanoquinone, sodium peroxodisulfate, ammonium peroxodisulfate, potassium peroxodisulfate, di-tert-butyl peroxide instead of silver acetate;

Methanol, ethanol, tetrahydrofuran, dichloromethane, chloroform, ether, benzene, o-dichlorobenzene, chlorobenzene, toluene, xylene, mesitylene, cyclohexane, petroleum ether, tert-amyl alcohol, 1,4 -Dioxane, 1,2-dichloroethane, N,N-dimethylformamide, N,N-dimethylacetamide instead of dimethylsulfoxide.

Through the screening of experimental parameters, it was found that under the above conditions, 3,5-dibenzimidazolyl-8-thienylfluoroborofluorine derivatives capable of specific fluorescence imaging and fluorescent labeling of the endoplasmic reticulum of cells could be successfully prepared. Among them, the product prepared in Example 1 has the best effect in fluorescence imaging and fluorescent labeling of endoplasmic reticulum.

The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements and improvements made within the spirit and principles of the present invention should be included in the protection of the present invention. within range.

Claims (9)

1. A 3,5-dibenzimidazolyl-8-thienylfluoroboron derivative, characterized in that it has the structure shown in the following formula I: Wherein: R ₁ , R ₂ , R ₃ are one or two of hydrogen, deuterium, alkyl, alkoxy, carbonyl, ester, halogen, substituted aryl or substituted heteroaryl. 2. A kind of 3,5-dibenzimidazolyl-8-thienylfluoroboron derivatives as claimed in claim 1, characterized in that, it has the structure shown in the following formula II: 3. The preparation method of a kind of 3,5-dibenzimidazolyl-8-thienyl fluoroboron derivatives as claimed in claim 1 or 2, characterized in that it comprises the following steps: 1) Mix 8-thienylfluoroborofluorine derivatives with benzimidazole derivatives, oxidant and solvent evenly, and then react under anhydrous and oxygen-free conditions. The reaction formula is as follows: 2) After the reaction is completed, cool to room temperature, remove the solvent and add dichloromethane to dissolve the reaction system, then filter through diatomaceous earth, and wash with dichloromethane, combine the filtrates, remove the solvent under reduced pressure, and use a silica gel column for the residue The compound of formula I can be obtained by separation and purification by chromatography and vacuum drying. 4. as claimed in claim 3, a kind of preparation method of 3,5-dibenzimidazolyl-8-thienyl fluorine boron derivatives is characterized in that, the oxidizing agent is copper acetate monohydrate, copper acetate, Copper chloride, copper bromide, ketone trifluoroacetate, copper (II) trifluoromethanesulfonate, copper acetylacetonate, silver carbonate, silver oxide, silver acetate, silver nitrate, silver hexafluoroantimonate, oxygen, iodobenzene acetate , benzoquinone, dichlorodicyanoquinone, sodium peroxodisulfate, ammonium peroxodisulfate, potassium peroxodisulfate, and di-tert-butyl peroxide. 5. as claimed in claim 3, a kind of preparation method of 3,5-dibenzimidazolyl-8-thienyl fluorine boron derivatives, is characterized in that, the solvent is methanol, ethanol, tetrahydrofuran, dichloro Methane, chloroform, ether, dimethyl sulfoxide, benzene, o-dichlorobenzene, chlorobenzene, toluene, xylene, mesitylene, cyclohexane, petroleum ether, tert-amyl alcohol, 1,4-diox One or more of hexacyclo, 1,2-dichloroethane, N,N-dimethylformamide, N,N-dimethylacetamide. 6. A kind of preparation method of 3,5-dibenzimidazolyl-8-thienyl fluorine fluoride derivatives as claimed in claim 3, characterized in that the reaction concentration of the 8-thienyl fluorine fluorine 0.0001～10mol/L. 7. The preparation method of a kind of 3,5-dibenzimidazolyl-8-thienyl fluorine fluorine derivatives as claimed in claim 3, characterized in that, the 8-thienyl fluorine fluorine derivatives , benzimidazole derivatives, the molar ratio of the oxidizing agent is 1:(0.01-50):(0.01-100). 8. A method for preparing a 3,5-dibenzimidazolyl-8-thienylfluoroboron derivative as claimed in claim 3, characterized in that the reaction temperature in step 1) is -40 to 160°C, the time ranges from 0.1 to 720 hours. 9. The application of a kind of 3,5-dibenzimidazolyl-8-thienyl fluorine boron derivatives as claimed in claim 1 or 2, characterized in that, its specific fluorescence imaging in endoplasmic reticulum and fluorescent labeling applications.