CN106188003B - Bis- target spot fluorescence probes of Cu2+ and Fe3+ based on chinoline backbone and its preparation method and application - Google Patents
Bis- target spot fluorescence probes of Cu2+ and Fe3+ based on chinoline backbone and its preparation method and application Download PDFInfo
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Abstract
The present invention provides a kind of Cu based on chinoline backbone2+And Fe3+Double target spot fluorescence probes and its preparation method and application, double target spot fluorescence probes have structure shown in Formulas I.Double target spot fluorescence probes of the invention are to Cu2+And Fe3+Identification have it is highly sensitive and highly selective.
Description
Technical field
The present invention relates to biological fluorescent labeling fields, more particularly, to a kind of Cu based on chinoline backbone2+And Fe3+
Double target spot fluorescence probes and its preparation method and application.
Background technique
Copper and iron are the metals being widely used in human production life, and the two has significant biology as transition elements
Correlation.They participate in the growth metabolism of organism often as coenzyme or other confactors.Excessive Cu2+And Fe3+It can draw
Play organism metabolism obstacle and other serious epidemic diseases.Cu2+And Fe3+Caused dysbolism disease mainly includes Organ Dysfunction
Or neurodegenerative disease.Therefore particularly significant for the detection of transition metal.
Fluorescence detection has the advantages such as highly sensitive, highly selective, the simple, quick response of operation.Cell fluorescence imaging is made
It is concerned for efficient detection means a kind of in biology and pharmaceutical science field.And it is used for the fluorescent material of cell imaging, it answers
When having good water solubility, high brightness, photostability and hypotoxicity.Fluorescent small molecule is due to its good biocompatibility, excellent
Photophysical Behaviors more are applied to intracellular image checking.Quinoline is a big conjugated system, can easily generate π
To the electron transition of π *, quinoline radical derivative not only has stronger fluorescence, but also its heterocyclic nitrogen atom can participate in being coordinated, i.e., together
When have identification and fluorescent functional.It is based on chinoline backbone specific recognition Zn in recent years2+, Ag+, Fe3+, Cu2+Plasma it is small
Fluorescence probe is by wide coverage.However, the fluorescence probe of the double target spots of identification simultaneously or multiple target point is by less report.Double target spots
Fluorescence probe has efficient, low cost, the easy advantage of operation, and therefore, double target spots, biofacies can be identified simultaneously by designing one kind
The good fluorescence probe of capacitive, which seems, to become more and more important.
Summary of the invention
It is an object of the invention to provide a kind of Cu based on chinoline backbone2+And Fe3+Double target spot fluorescence probes, and providing should
The preparation method of double target spot fluorescence probes and its application in cell imaging.
The first aspect of the present invention is to provide a kind of Cu based on chinoline backbone2+And Fe3+Double target spot fluorescence probes are (following
Abbreviation QLBM), which has structure shown in Formulas I:
The second aspect of the present invention is to provide the preparation method of double target spot fluorescence probes, this method comprises:
(i) it in the presence of the first organic solvent, reacts quinaldinic acid with oxalyl chloride, obtains compound shown in Formula II；
(ii) it in the presence of the second organic solvent, reacts compound shown in Formula II with 2- aminobenzimidazole, obtains Formulas I
Shown compound.
The third aspect of the present invention is to provide the application of double target spot fluorescence probes in cell imaging.
Effect of the invention is shown:
(1) QLBM is to Cu2+And Fe3+Identification have it is highly sensitive and highly selective.
(2) reaction system of the probe is pure water phase.
(3) it is put forward for the first time using ascorbic acid to Cu2+And Fe3+It distinguishes.
(4) its mechanism of action is analyzed using mass spectrum, DFT, and as Cu2+And Fe3+Double target spot probe applications are in thin
Born of the same parents' imaging.
Other features and advantages of the present invention will then part of the detailed description can be specified.
Detailed description of the invention
Exemplary embodiment of the invention is described in more detail in conjunction with the accompanying drawings, it is of the invention above-mentioned and its
Its purpose, feature and advantage will be apparent.
Fig. 1 is the hydrogen spectrum of the double target spot fluorescence probes of the present invention.
Fig. 2 is the carbon spectrum of the double target spot fluorescence probes of the present invention.
Fig. 3 is the UV absorption spectrogram of the double target spot fluorescence probes of the present invention.
Fig. 4 (a) shows Tris-HCl (50mM, the pH that 200 μM of different metal ions are added to QLBM (20 μM)
7.2) fluorescent emission spectrogram after solution；Fig. 4 (b), which is shown, is added QLBM (20 μM) for 200 μM of different metal ions
Color diagram after Tris-HCl (50mM, pH 7.2) solution under ultraviolet light.Fig. 4 (c) and Fig. 4 (d) respectively illustrate QLBM (20 μ
M) in Tris-HCl (50mM, pH 7.2) solution with Cu2+(200 μM), Fe3+(200 μM) and other metal ions (500 μM)
Fluorescent emission spectrogram.Secret note represents other competition metal ions.Red item, which is represented, is added Cu after competition metal ion is added2+
(200 μM) and Fe3+(200μM)。
Fig. 5 (a), which is shown, is constantly added dropwise Cu in Tris-HCl (50mM, pH 7.2) solution of QLBM (20 μM)2+It is glimmering
Light emitting spectrogram.Fig. 5 (b), which is shown, is constantly added dropwise Cu in Tris-HCl (50mM, pH 7.2) solution in QLBM (20 μM)2+
Fluorescent value and Cu at 508nm2+The titration curve of concentration is added dropwise.Inner curve is QLBM fluorescent value and Cu2+Ion concentration
Linearly interval curve.Fig. 5 (c), which is shown, is constantly added dropwise Fe in Tris-HCl (50mM, pH 7.2) solution of QLBM (20 μM)3+
Fluorescent emission spectrogram.Fig. 5 (d) is shown constantly to be added dropwise in Tris-HCl (50mM, pH 7.2) solution in QLBM (20 μM)
Fe3+Fluorescent value and Fe at 508nm3+The titration curve of concentration is added dropwise.Inner curve is QLBM fluorescent value and Fe3+Ion is dense
The linearly interval curve of degree.
Fig. 6 (a), which is shown, is added Fe in Tris-HCl (50mM, pH 7.2) solution of QLBM (20 μM)3+(200 μM) from
After son, and the fluorescent emission spectrogram variation after ascorbic acid (500 μM) is added.Fig. 6 (b) is shown QLBM's (20 μM)
Cu is added in Tris-HCl (50mM, pH 7.2) solution2+After (200 μM) ion, and it is added glimmering after ascorbic acid (500 μM)
The variation of light emitting spectrogram.Fig. 6 (c) is shown comprising Cu2+(200 μM) and Fe3+The Tris- of the QLBM (20 μM) of (200 μM)
Be added in HCl (50mM, pH 7.2) solution ascorbic acid (500 μM) afterwards at maximum emission wavelength (508nm) time gradient it is glimmering
Light value variation.Fig. 6 (d), which is shown, is added Cu in Tris-HCl (50mM, pH 7.2) solution of QLBM (20 μM)2+(200μM)
And Fe3+The color change under ascorbic acid (500 μM) ultraviolet light irradiation is added in (200 μM) afterwards.
Fig. 7 shows QLBM-CuCl2With 2 equivalent QLBM-FeCl3Mass spectral results.
Fig. 8 (a), Fig. 8 (b), Fig. 8 (c) respectively illustrate QLBM, QLBM-Cu that DFT calculates analysis2+And QLBM-Fe3+'s
Optimization structure.
Fig. 9 shows the cytotoxicity of various concentration QLBM.
Figure 10 (a) is that HeLa cell and 20 μM of QLBM are incubated for the Laser Scanning Confocal Microscope imaging after 6h altogether.Figure 10 (b) is
After HeLa cell and 20 μM of QLBM are incubated for 6h altogether, 200 μM of Cu are added2+It is incubated for the Laser Scanning Confocal Microscope imaging of 30min.Figure 10
(c) after being incubated for 6h altogether for HeLa cell and 20 μM of QLBM, 200 μM of Fe are added3+It is incubated for the Laser Scanning Confocal Microscope imaging of 30min.
Scale: 50 μm.
Specific embodiment
The present invention will be described in more detail below with reference to accompanying drawings.
The present invention provides a kind of Cu based on chinoline backbone2+And Fe3+Double target spot fluorescence probes, double target spot fluorescence probes
With structure shown in Formulas I:
The present invention provides the preparation method of above-mentioned double target spot fluorescence probes, this method comprises:
(i) it in the presence of the first organic solvent, reacts quinaldinic acid with oxalyl chloride, obtains compound shown in Formula II；
(ii) it in the presence of the second organic solvent, reacts compound shown in Formula II with 2- aminobenzimidazole, obtains Formulas I
Shown compound.
In accordance with the present invention it is preferred that the molar ratio of quinaldinic acid and oxalyl chloride is 1:5-20 in step (i).
First organic solvent in step (i) can be the aprotic organic solvent of this field routine, preferably two
At least one of chloromethanes, toluene and n-hexane.It is further preferred that above-mentioned organic solvent is organic molten after being dried
Agent.
Specifically, step (i) can include:
The oxalyl chloride being dissolved in the first organic solvent is added drop-wise in quinaldinic acid solution, after mixing, in inertia
In the presence of gas, distillation is stirred to react.
Wherein, the quinaldinic acid solution is preferably that quinaldinic acid is dissolved in the solution formed in the first organic solvent, quinoline
The concentration of which pyridine acid solution can according to need determination.
The dropwise addition preferably carries out at low temperature, such as condition of ice bath.
The inert gas for example can be nitrogen.
Step (i) can also include conventional post-processing step, such as cooling, revolving solvent etc..
According to the present invention, in step (ii), the molar ratio of compound shown in Formula II and 2- aminobenzimidazole is preferably 1:
1-2。
Second organic solvent in step (ii) can be the aprotic organic solvent of this field routine, preferably two
At least one of chloromethanes, toluene and n-hexane.It is further preferred that above-mentioned organic solvent is organic molten after being dried
Agent.
Preferably, step (ii) includes: that acid binding agent is added into reaction system.The acid binding agent for example can be trimethylamine
Or triethylamine.
According to the present invention, the reaction condition of step (ii) preferably includes: temperature is -4 to 5 DEG C, time 2-10h.
Step (ii) can also include conventional post-processing step, such as cooling, revolving solvent, column chromatography etc..
The present invention also provides application of the above-mentioned double target spot fluorescence probes in cell imaging.
A kind of preferred embodiment according to the present invention has synthesized fluorescence probe QLBM by two-step reaction design, had synthesized
Journey is shown below:
Synthetic method: quinaldinic acid being dissolved in dry methylene chloride, and oxalyl chloride is then dissolved in dry dichloromethane
In alkane, under condition of ice bath and magnetic agitation, oxalyl chloride solution is added dropwise in quinaldinic acid solution, later in a nitrogen environment
Distillation reaction.Then, reaction solution is cooled to room temperature, revolving removes solvent, obtains crude product quinaldine acyl chlorides (referred to as C2).
Then obtained product C2 and 2- aminobenzimidazole are added in the mixed solution of dry methylene chloride and trimethylamine,
It is stirred to react, revolving removes solvent, and by silica gel column chromatography separating purification, obtains QLBM.
By following embodiment, invention is further explained.
Embodiment 1
Quinaldinic acid (C1,0.38g, 2mM) is dissolved in the two-mouth bottle that dry methylene chloride (20mL) is added to 100mL
In, it is put into magnetic rotor.Then oxalyl chloride (2.60g, 20mM) is dissolved in the dry methylene chloride of 10mL, in ice bath item
It is added dropwise in quinaldinic acid solution, stirs 30 minutes, 5 hours are stirred in distillation in a nitrogen environment later under part.It then, will be anti-
Liquid is answered to be cooled to room temperature, revolving removes solvent, obtains crude product quinaldine acyl chlorides (referred to as C2).Then the product C2 that will be obtained
It is added to 2- aminobenzimidazole (0.27g, 2mM) molten with the mixing of the trimethylamine of 0.1mL in the dry methylene chloride of 30mL
In liquid, 5 hours are stirred to react under the conditions of 0 DEG C, revolving removes solvent, and by silica gel column chromatography separating purification, obtains QLBM
Yellow solid 0.42g, yield 73%.
Fig. 1 and Fig. 2 is respectively the hydrogen spectrum and carbon spectrum of QLBM.
Test case 1
The optical physics of 1 QLBM characterizes
The absorption spectrogram of 1.1 test QLBM.Tris-HCl (50 mM, pH 7.2) solution of the QLBM of 20 μM of configuration, takes
2mL is added in quartz colorimetric utensil, tests it using ultraviolet-visible spectrophotometer and absorbs spectrogram.
The fluorescent emission spectrogram of 1.2 test QLBM.The QLBM solution of 20 μM of configuration in 50mM Tris-HCl (pH 7.2),
It takes 1mL to be added in cuvette, the fluorescent emission spectrogram of QLBM is tested on sepectrophotofluorometer.Excitation wavelength is 370nm,
Transmitting section is 400-700nm.
Test case 2
Selection Journal of Sex Research of 2 QLBM to different metal ions
The aqueous solution of the Tris-HCl (50mM, pH 7.2) of 2.1 configurations QLBM (20 μM), is added into each cuvette
The solution of 1mL sequentially adds Na in each cuvette+, K+, Mg2+, Zn2+, Ca2+, Fe2+, Mn2+, Hg2+, Cd2+, Co2+, Al3+,
Pd2+, Cu2+, Fe3+Salting liquid.It is uniformly mixed after addition, concentration of metal ions is Na in each ware+(500 μM), K+(500 μM),
Mg2+(500 μM), Zn2+(500 μM), Ca2+(500 μM), Fe2+(500 μM), Mn2+(500 μM), Hg2+(500 μM), Cd2+(500μ
M), Co2+(500 μM), Al3+(500 μM), Pd2+(500 μM) ion salt solution.
2.2 carry out the detection (excitation wavelength 370nm) of fluorescence intensity on sepectrophotofluorometer.
2.3 acquire and handle data.
Test case 3
3 Cu2+And Fe3+To the titration experiments of QLBM
3.1 20 μM of QLBM solution of configuration, solvent are 50mM Tris-HCl (pH 7.2), and 1mL is taken to be added in cuvette.
The CuCl of 3.2 configuration 10mM2And FeCl3Aqueous solution.
3.3 Cu is added dropwise into cuvette respectively2+, Cu2+Concentration range be 0~300 μM, on sepectrophotofluorometer
Successively test fluorescent emission spectrogram.
Fe is added dropwise in 3.4 same methods, test3+Fluorescent emission spectrogram.
3.5 acquire and handle data.
Test case 4
4 ascorbic acid are to Cu2+And Fe3+Differentiation
4.1 20 μM of QLBM solution of configuration, solvent are 50mM Tris-HCl (pH 7.2), and 1mL is taken to be separately added into two ratios
In color ware.
4.2 are separately added into 200 μM of Cu2+And Fe3+Salting liquid tests fluorescent emission spectrogram.
4.3 are then separately added into 500 μM of ascorbic acid solutions, are uniformly mixed, the fluorescent emission map in record 1h, and every 10
Minute test is primary.
4.4 acquire and handle data.
Test case 5
5 QLBM and Cu2+And Fe3+Mechanism of action analysis
The CuCl of 5.1 configuration QLBM and two equivalents2And FeCl3Solution, be analyzed by mass spectrometry.
5.2 mass spectral analyses obtain QLBM and Cu2+/Fe3+In conjunction with ratio.
5.3, which carry out density functional theory (DFT) using 09 software of Gauss on computers, calculates analysis.
Test case 6
6 cell culture processes
HeLa cell is placed in 37 DEG C, carbon dioxide content is in 5% incubator.Culture medium is containing 10%FBS's
RPMI-1640.It is passed on when cell density is about 90% using pancreatin cell dissociation buffer, passage in average 2 days is primary.
Test case 7
The detection of 7 cytotoxicities
Logarithmic phase HeLa cell is collected, adjustment cell solution concentration to 50000/mL, every hole is added 100 in 96 orifice plates
μL.In 5%CO2, after 37 DEG C of incubator overnight incubation, the 100 μ L of QLBM of various concentration gradient is added, makes each gradient concentration
For (0,2 μM, 5 μM, 10 μM, 20 μM, 50 μM, 100 μM).Five multiple holes of each gradient.After culture for 24 hours, every hole is added 20 μ L's
3- (4,5- dimethylthiazole -2) -2,5- diphenyltetrazolium bromide bromide (MTT) PBS solution (5mg/mL), is incubated for 4h altogether.It inhales
Supernatant is removed, the DMSO of 150 μ L is added in every hole, sets low speed on shaking table and shakes 10 minutes, dissolves crystal sufficiently.In enzyme linked immunological
The light absorption value in each hole is measured at detector OD 490nm.
Test case 8
8 cell imagings
8.1 QLBM and HeLa cytosis imaging experiment.HeLa cell is passaged to the copolymerization coke glass that diameter is 20mm
Bottom culture dish, in 5%CO2, after 37 DEG C of incubator culture for 24 hours.QLBM is added in culture medium, the concentration of QLBM is 20 μM,
After being incubated for 6h, culture solution is removed, is cleaned cell six times with 1 × PBS.In Laser Scanning Confocal Microscope (Olympus FV1000-IX81
Confocal laser scanning microscope) under observe.Exciting light selects mercury laser (72.0%405nm).
8.2 intracellular detection QLBM and Cu2+And Fe3+Cell imaging experiment.It is 20mm's that HeLa cell, which is passaged to diameter,
It is copolymerized burnt Glass bottom culture dish, in 5%CO2, after 37 DEG C of incubator culture for 24 hours.QLBM is added in culture medium, QLBM's is dense
Degree is 20 μM, and after being incubated for 6h, 200 μM of Cu are added2+And Fe3+Culture solution is removed after being incubated for 30min altogether, cleans cell with 1 × PBS
Six times.At Laser Scanning Confocal Microscope (Olympus FV1000-IX81 confocal laser scanning microscope)
Observation.Exciting light selects mercury laser (72.0%405nm).
9 data and analysis
UV absorption and fluorescent emission spectrogram of 9.1 QLBM in Tris-HCl (50mM, pH 7.2) solution system
As shown in figure 3, QLBM possesses the absorption region of 300~400nm.It is its maximum emission wavelength at 508nm.
The selection Journal of Sex Research of 9.2 QLBM and different metal ions
QLBM is designed based on chinoline backbone, and the quinoline of heterocyclic nitrogen atom can be used as the chela of metal ion
Coincidence point, so carrying out the selection Journal of Sex Research between QLBM and different metal ions, the present invention has chosen 14 kinds of common metals altogether
Ion (Na+, K+, Mg2+, Zn2+, Ca2+, Fe2+, Mn2+, Hg2+, Cd2+, Co2+, Al3+, Pd2+, Cu2+, Fe3+)。
As shown in Fig. 4 (a) -4 (d), after joined different metal ions, Cu2+And Fe3+The fluorescence display of QLBM is gone out
It is significantly quenched, and other ions is not changed significantly.It is corresponding to it, QLBM-Cu under ultraviolet light irradiation2+With
QLBM-Fe3+Fluorescence have and be significantly quenched, and the not apparent color change of the QLBM solution that other ions are added.Other from
Cu is then separately added into the QLBM solution of son2+And Fe3+, the QLBM solution fluorescence of other ions shows and is significantly quenched.With
Upper result illustrates QLBM to Cu2+And Fe3+There is a special response, and under the conditions of existing for other ions, QLBM will not be interfered to Cu2 +And Fe3+Special response.
9.3 Cu2+And Fe3+To the titration experiments of QLBM
As shown in Fig. 5 (a) -5 (d), in Tris-HCl (50mM, pH 7.2) solution system of QLBM (20 μM), with
Cu2+And Fe3+Dropwise addition concentration be continuously increased, the fluorescent value at the maximum emission wavelength (508nm) of QLBM constantly declines, and works as Cu2+
And Fe3+Dropwise addition concentration when reaching 200 μM, the fluorescent value of QLBM is quenched efficiency and reaches about 90%.Work as Cu2+And Fe3+Dropwise addition
Concentration is at 0~50 μM, the maximum emission wavelength (508nm) and Cu of QLBM2+/Fe3+Dropwise addition concentration present it is preferable linear.
Its detection line is calculated by LOD=3*Sb/S, QLBM is to Cu2+Detection be limited to 1.35 × 10-7M, to Fe3+Detection be limited to
1.24×10-7M.In Tris-HCl (50mM, pH 7.2) solution system of QLBM (20 μM), with Cu2+And Fe3+Dropwise addition
Concentration is continuously increased, and the fluorescent value at the maximum emission wavelength (508nm) of QLBM constantly declines, and works as Cu2+And Fe3+Dropwise addition it is dense
When degree reaches 200 μM, the fluorescent value of QLBM is quenched efficiency and reaches about 90%.Work as Cu2+And Fe3+Dropwise addition concentration at 0~50 μM
When, the maximum emission wavelength (508nm) and Cu of QLBM2+/Fe3+Dropwise addition concentration present it is preferable linear.Pass through LOD=3*Sb/
S calculates its detection line, and QLBM is to Cu2+Detection be limited to 1.35 × 10-7M, to Fe3+Detection be limited to 1.24 × 10-7M。
9.4 ascorbic acid (AA) are to Cu2+And Fe3+Differentiation
As shown in Fig. 6 (a) -6 (d), Cu2+And Fe3+To QLBM all show preferable quenching effects (be quenched rate~
90%) after excessive ascorbic acid (500 μM), are then added, QLBM-Fe3+Solution shows that apparent fluorescence is replied, and
QLBM-Cu2+Fluorescence have no significant changes.This is mainly due to ascorbic acid to have stronger reproducibility, can be quickly by Fe3 +It is reduced to Fe2+.According to Fig. 4 the results show that QLBM is to Fe2+Do not obvious response to.And ascorbic acid is to Cu2+Reduction need
More harsh condition, thus QLBM-Cu2+Fluorescence have no significant changes.Under ultraviolet light irradiation solution colour variation also with it is upper
It is consistent to state result.Ascorbic acid and Fe3+Reaction belong to redox reaction, which needs the regular hour, but investigate
Ascorbic acid and Fe3+Action time, find after reaching 1h, fluorescence is returned to maximum value.
9.5 QLBM and Cu2+And Fe3+Study on mechanism
The mass spectral results of Fig. 7 (a) -7 (b) show that m/z=289.1106 (calculated value=289.1084) corresponds to [QLBM+
H]+, m/z=350.0238 (calculated value=350.0218) is corresponding to [QLBM+Cu2++ H]+, m/z=385.9989 (calculated value
=385.9990) correspond to [QLBM+Cu2++Cl-]+, m/z=378.0009 (calculated value=378.0410) correspond to [QLBM+
Fe3++Cl-+OH-]+, m/z=396.0105 (calculated value=396.0071) is corresponding to [QLBM+Fe3++2OH-]+, m/z=
413.9745 (calculated value=413.9732) correspond to [QLBM+Fe3++2Cl-]+.QLBM and Cu as the result is shown2+And Fe3+Knot
Composition and division in a proportion example is 1:1.
It is calculated and is analyzed by DFT, QLBM, QLBM and Cu2+And QLBM and Fe3+Binding site such as Fig. 8 (a), 8 (b) and figure
Shown in 8 (c).Cu2+Distance with the N (1) in QLBM structure, N (2), Cl (3), Cl (4) atom is respectivelyWithFe3+With the N (1) in QLBM structure, N (2), Cl (3), Cl (4) atom
Distance is respectively WithStructure is QLBM in figure,
QLBM-Cu2+Compound and QLBM-Fe3+The optimization structure of compound.
9.6 cell imaging
9.6.1 cytotoxicity
The cytotoxicity of QLBM is studied by mtt assay, as shown in figure 9, after being incubated for for 24 hours when QLBM concentration reaches 20 μM,
Cell survival rate is up to 90% or more.
9.6.2 the intracellular imaging of QLBM
It is in Figure 10 (a) -10 (c) the results show that carry out confocal microscopic image after 6h after HeLa cell is incubated for altogether,
HeLa cell shows bright green fluorescence.After being incubated for 6h, it is separately added into 200 μM of Cu2+Ion and 200 μM of Fe3+Ion is laggard
Row confocal microscopic image, the fluorescence presentation compared with Figure 10 (a) of HeLa cell are significantly quenched.QLBM can be used as the result is shown
Make intracellular image checking Cu2+And Fe3+。
Claims (9)
1. pair target spot fluorescence probe is preparing the application in cell imaging reagent, which is characterized in that double target spot fluorescence probes are
Cu2+And Fe3+Double target spot fluorescence probes, with structure shown in Formulas I:
2. application according to claim 1, which is characterized in that double target spot fluorescence probes, which pass through, to be included the following steps
Method is made:
(i) it in the presence of the first organic solvent, reacts quinaldinic acid with oxalyl chloride, obtains compound shown in Formula II；
(ii) it in the presence of the second organic solvent, reacts compound shown in Formula II with 2- aminobenzimidazole, obtains shown in Formulas I
Compound.
3. application according to claim 2, wherein in step (i), the molar ratio of quinaldinic acid and oxalyl chloride is 1:5-
20。
4. application according to claim 2, wherein first organic solvent is in methylene chloride, toluene and n-hexane
At least one.
5. application according to claim 2, wherein step (i) includes:
The oxalyl chloride being dissolved in the first organic solvent is added drop-wise in quinaldinic acid solution, after mixing, in inert gas
In the presence of, distillation is stirred to react.
6. application according to claim 2, wherein in step (ii), compound shown in Formula II and 2- aminobenzimidazole
Molar ratio be 1:1-2.
7. application according to claim 2, wherein second organic solvent is in methylene chloride, toluene and n-hexane
At least one.
8. application according to claim 2, wherein step (ii) includes: that acid binding agent is added into reaction system.
9. application according to claim 2, wherein the reaction condition of step (ii) includes: that temperature is -4 to 5 DEG C, the time
For 2-10h.
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