Fluoroboron fluorescent dye derivatives and preparation methods and applications thereof

Abstract

The present invention discloses a fluoroboron fluorescent dye derivative. The structure of the fluoroboron fluorescent dye derivative is shown in formula (III), wherein R1 is an aryl group or a cycloalkyne, and R2 is an aryl group or a substituted aryl group. The fluoroboron fluorescent dye derivative has excellent fluorescence quantum yield, solubility and stability. The preparation method has the advantages of simple steps and readily available raw materials, thereby enabling the fluoroboron fluorescent dye derivative to be widely used in fluorescent labeling.

Description

Technical Field

[0001] The present invention relates to the field of fluorine-boron fluorescent dyes, and in particular to a fluorine-boron fluorescent dye derivative and a preparation method and application thereof. Background Art

[0002] Boron dipyrrole (BODIPY) fluorescent dyes, a class of molecules developed only in the past two decades, exhibit excellent photophysical and chemical properties. They possess narrow absorption and emission peaks, high molar absorptivity, high fluorescence quantum yield, and excellent photostability and chemical stability. However, traditional BODIPY fluorescent dyes have certain limitations in their applications, such as their relatively small Stokes shift and susceptibility to fluorescence quenching.

[0003] Therefore, it is of great significance to prepare a new fluoroboron fluorescent dye with high fluorescence quantum yield and good solubility. At the same time, the existing methods for synthesizing new BODIPY fluorescent dye analogs either have complicated steps or raw materials are difficult to obtain, require multiple steps, are difficult to synthesize, and have low yields. Therefore, it is also of great significance to design a method for preparing fluorescent dyes with simple steps and readily available raw materials. Summary of the Invention

[0004] The purpose of the present invention is to provide a fluoroboron fluorescent dye derivative and its preparation method and application. The fluoroboron fluorescent dye derivative has excellent fluorescence quantum yield, solubility and stability. At the same time, the preparation method has the advantages of simple steps and readily available raw materials, thereby enabling the fluoroboron fluorescent dye derivative to be widely used in fluorescent labeling.

[0005] In order to achieve the above object, the present invention provides a fluoroboron fluorescent dye derivative, the structure of which is shown in formula (III):

[0006]

[0007] Wherein, R1 is an aryl group or a cycloalkadiene group, and R2 is an aryl group or a substituted aryl group.

[0008] The present invention also provides a method for preparing the fluoroboron fluorescent dye derivative as described above, wherein the method comprises: in the presence of a catalyst, contacting a fluoroboron fluorescent dye having a structure represented by formula (VIII) with a 2-(tributyltinyl) compound in an organic solution to obtain the fluoroboron fluorescent dye derivative represented by formula (III).

[0009]

[0010] In formula (VIII), R2 is an aryl group or a substituted aryl group.

[0011] The present invention further provides a use of the above-mentioned fluoroboron fluorescent dye derivative in fluorescent labeling.

[0012] In the above technical scheme, the fluoroboron fluorescent dye derivative prepared by the present invention has a maximum fluorescence emission wavelength between 494-505 nm, has broad absorption and emission peaks, excellent fluorescence quantum yield, excellent solubility, and excellent stability, and has potential for wide application in fluorescent labeling; at the same time, the preparation method of the fluoroboron fluorescent dye derivative with the structure shown in formula (III) provided by the present invention has simple steps and the raw materials are easily available.

[0013] Other features and advantages of the present invention will be described in detail in the following detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. Together with the following detailed description, they are used to explain the present invention but do not constitute a limitation of the present invention. In the accompanying drawings:

[0015] Figure 1 These are the front and side views of the crystal structures of fluoroboron fluorescent dye derivatives C4, C6, C9, C7, and C8, respectively. DETAILED DESCRIPTION

[0016] The following is a detailed description of the specific embodiments of the present invention. It should be understood that the specific embodiments described herein are only used to illustrate and explain the present invention and are not intended to limit the present invention.

[0017] The endpoints of the ranges and any values ​​disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoints of each range, the endpoints of each range and individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered to be specifically disclosed herein.

[0018] The present invention provides a fluoroboron fluorescent dye derivative, the structure of which is shown in formula (III):

[0019]

[0020] Wherein, R1 is an aryl group or a cycloalkadiene group, and R2 is an aryl group or a substituted aryl group.

[0021] In the above-mentioned fluoroboron fluorescent dye derivatives, the specific types of each substituent can be selected within a wide range, but considering the difficulty of sourcing the raw materials and the yield of the synthesis, preferably, R1 is phenyl, sulfur-substituted cyclopentadiene or oxygen-substituted cyclopentadiene, and R2 is halogen-substituted phenyl, nitro-substituted phenyl, phenyl, or alkyl-substituted phenyl; more preferably, R1 is phenyl, thienyl or furyl, and R2 is 2,6-dichlorophenyl, 2,4,6-trimethylphenyl, phenyl or p-nitrophenyl.

[0022] In the above-mentioned fluoroboron fluorescent dye derivatives, in combination with actual conditions, preferably, the structure of the fluoroboron fluorescent dye derivative is as shown in Formula C1-C9,

[0023]

[0024] The present invention also provides a method for preparing the fluoroboron fluorescent dye derivative as described above, wherein the method comprises: in the presence of a catalyst, contacting the fluoroboron fluorescent dye of the structure represented by formula (VIII) with a 2-(tributyltinyl) compound in an organic solution to react;

[0025]

[0026] In formula (VIII), R2 is an aryl group or a substituted aryl group.

[0027] In the above formulae, the specific type of the substituent can be selected from a wide range, but considering the difficulty and cost of preparation, preferably, R2 is a halogen-substituted phenyl group, a nitro-substituted phenyl group, a phenyl group, or an alkyl-substituted phenyl group;

[0028] More preferably, R2 is 2,6-dichlorophenyl, 2,4,6-trimethylphenyl, phenyl, or p-nitrophenyl.

[0029] In the above embodiment, the amount of each material can be selected within a wide range, but in order to further improve the yield, preferably, the molar ratio of the compound represented by the structure of formula (VIII), the catalyst, and the 2-(tributyltinyl) reagent is 0.2:0.2-0.5:0.3-0.5;

[0030] More preferably, the molar ratio of the compound represented by formula (VIII), the catalyst, and the 2-(tributyltinyl) reagent is 0.2:0.3-0.4:0.4-0.5.

[0031] In the present invention, the specific type of catalyst can be selected within a wide range, but in order to further improve the catalytic efficiency, preferably, the catalyst is a metal chloride;

[0032] More preferably, the catalyst is selected from aluminum chloride and / or copper chloride;

[0033] More preferably, the catalyst is aluminum trichloride.

[0034] In the present invention, in order to further increase the reaction rate and improve the yield of the product, preferably, the preparation method is carried out in an organic solvent, and the organic solvent is selected from at least one of chlorobenzene, dichloromethane, and toluene;

[0035] More preferably, the organic solvent is selected from dichloromethane and / or toluene.

[0036] In the above embodiment, the conditions for the contact reaction can be selected within a wide range. However, in order to further improve the yield, preferably, the conditions for the contact reaction include: a reaction temperature of 20-30° C., and a reaction time of 1-3 h.

[0037] The present invention further provides a use of the above-mentioned fluoroboron fluorescent dye derivative in fluorescent labeling.

[0038] The present invention will be described in detail below through examples.

[0039] The nuclear magnetic resonance measurements were performed using an AV-500 nuclear magnetic resonance instrument from Bruker, Switzerland; the mass spectra were performed using an HPLC / ESI-MS mass spectrometer from the American Instrument Group; the ultraviolet spectra were performed using a UV-2450 ultraviolet / visible spectrophotometer from Shimadzu, Japan; the fluorescence spectra were performed using an F-4500FL fluorescence spectrophotometer from Hitachi, Japan; the relative fluorescence quantum yield was performed using an F-4500FL fluorescence spectrophotometer from Hitachi, Japan; and the single crystal diffraction was performed using a SMAR APEXⅡX-single crystal diffractometer from Bruker AXS, Germany. max represents the maximum absorption wavelength, ε abs represents the molar extinction coefficient, λ em max represents the maximum fluorescence emission wavelength, Φ F Relative fluorescence quantum yield (Φ) and Stokes-shift (Φ) represent Stokes shift. F ) is determined by the relative fluorescence quantum yield Φ F The determination of fluorescent yellow (Φ = 0.90, in 0.1 mol / L sodium hydroxide solution) is used as the standard dye, according to the formula Φ F =Φ S *(I X / I S )*(A S / A X )*(n X / n S ) 2 Calculated, where ΦS is the fluorescence quantum yield of the standard CV, I is the integrated area of ​​the spectrum, A is the absorbance, n is the refractive index of the solvent, the subscript S is the standard, and X is the analyte.

[0040] The raw materials used in the following examples: dichloromethane, toluene, aluminum chloride, pyrrole, and trifluoroacetic acid are products of Sinopharm Chemical Reagent Co., Ltd., DDQ and 2-(tributyltin alkyl) aromatic reagent are products of Anaiji Chemical Co., Ltd.; the fluoroboron dipyrrole shown in formula (VIII) is one of the compounds with structures shown in formulas B1-B5.

[0041]

[0042] Example 1

[0043] Preparation of the fluoroboron fluorescent dye derivative represented by formula C1:

[0044]

[0045] 62 mg (0.2 mmol) of fluoroboron dipyrrole B1 represented by formula (VIII) was weighed and placed in a dry Schlenk reaction flask under argon protection, followed by the addition of 6 mL of toluene. After B1 was completely dissolved under stirring, 40 mg (0.3 mmol, 1.5 equiv) of aluminum trichloride was added. After stirring at room temperature for 20 minutes, tributylphenyltin (0.14 mL, 2.2 equiv) was added. After reacting at 80°C for 2 hours, the reaction mixture was dried using a rotary evaporator and separated by column chromatography (dichloromethane / petroleum ether = 1 / 4) to obtain a yellow solid C1 (yield 72%).

[0046] The characterization data are as follows: 1 H NMR (500MHz, CDCl3, ppm): δ7.55(d,J=1.0Hz,2H),7.35–7.13(m,10H),6.94(s,2H),6.74–6.65(m,2H),6.48–6.37(m,2H),2.36(s,3H),2.02(s,6H). 13 CNMR (125MHz, CDCl3, ppm): δ147.0,144.8,138.3,136.3,135.0,132.8,130.6,128.0,127.7,127.3,126.1,117.5,21.2,19.8.HRMS(APCI)Calcd.For C30H27BN2[M+H] + :427.2337,found 427.2346.

[0047] Example 2

[0048] Preparation of the fluoroboron fluorescent dye derivative represented by formula C2:

[0049]

[0050] 62 mg (0.2 mmol) of fluoroboron dipyrrole B1 represented by formula (VIII) was weighed into a dry Schlenk reaction flask under argon, followed by the addition of 6 mL of dichloromethane. After complete dissolution of B1 with stirring, 40 mg (0.3 mmol, 1.5 equiv) of aluminum trichloride was added. Stirring was continued at room temperature for 20 minutes, followed by the addition of 2-(tributylstannyl)thiophene (0.14 mL, 2.2 equiv). The reaction was monitored by thin-layer chromatography. After approximately 2 hours, the reaction mixture was dried using a rotary evaporator and isolated by column chromatography (dichloromethane / petroleum ether = 1 / 4) to afford C2 as a yellow solid (81% yield).

[0051] The characterization data are as follows: 1 H NMR (500MHz, CDCl3, ppm): δ7.71(s,2H),7.38(d,J=4.5Hz,2H),7.06(t,J=3.5Hz,2H),7.00(d,J= 3.0Hz,2H),6.95(s,2H),6.71(d,J=4.5Hz,2H),6.45(d,J=4.0Hz,2H),2.37(s,3H),2.07(s,6H). 13 CNMR (125MHz, CDCl3, ppm): δ147.0,145.5,138.5,136.4,134.5,130.5,130.4,128.3,128.0,127.4,127.0,117.9,21.2,19.8.HRMS(APCI)Calcd.For C 26 H 23 BN2S2[M+H] + :439.1466,found 439.1465.

[0052] Example 3

[0053] Preparation of the fluoroboron fluorescent dye derivative represented by formula C3:

[0054]

[0055] 62 mg (0.2 mmol) of fluoroboron dipyrrole B1 represented by formula (VIII) was weighed into a dry Schlenk reaction flask under argon, followed by the addition of 6 mL of dichloromethane. After complete dissolution of B1 with stirring, 40 mg (0.3 mmol, 1.5 equiv) of aluminum trichloride was added. Stirring was continued at room temperature for 20 minutes, followed by the addition of 2-(tributylstannyl)furan (0.14 mL, 2.2 equiv). The reaction was monitored by thin-layer chromatography. After approximately 2 hours, the reaction mixture was dried using a rotary evaporator and isolated by column chromatography (dichloromethane / petroleum ether = 1 / 3) to afford C3, a red solid (90% yield).

[0056] The characterization data are as follows: 1 H NMR (500MHz, CDCl3, ppm): δ7.72(s,2H),7.51(d,J=1.0Hz,2H),6.95(s,2H),6.70(d,J=3.0Hz,2H),6.4 5(dd,J=4.0,1.5Hz,2H),6.30(dd,J=3.0,1.5Hz,2H),6.18(d,J=3.0Hz,2H),2.36(s,3H),2.08(s,6H). 13 C NMR (125MHz, CDCl3, ppm): δ147.0,145.2,143.8,138.4,136.4,134.4,130.5,128.0,117.8,113.6,109.1,21.2,19.9.HRMS(APCI)Calcd.For C 26 H 23 BN2O2[M+H] + :407.1922,found 407.1931.

[0057] Example 4

[0058] Preparation of the fluoroboron fluorescent dye derivative represented by formula C4:

[0059]

[0060] The reaction was carried out in the same manner as in Example 2, except that the fluoroboron dipyrrole represented by formula (VIII) was replaced by B2 (54 mg, 0.2 mmol). Post-reaction treatment was the same as in Example 2. Column chromatography (dichloromethane / petroleum ether = 1 / 4) was used to separate the yellow solid C4 (yield 89%).

[0061] The characterization data are as follows: 1H NMR (500MHz, CDCl3, ppm): δ7.76 (s, 2H), 7.63–7.50 (m, 5H), 7.38 (d, J = 5.0Hz, 2H), 7.0 5(t,J=3.5Hz,2H),6.98(d,J=4.0Hz,2H),6.94(d,J=3.0Hz,2H),6.54(d,J=3.0Hz,2H). 13 C NMR (125MHz, CDCl3, ppm): δ145.6,134.3,134.2,130.6,130.5,130.3,129.8,128.2,127.4,127.1,126.9,125.1,117.9.HRMS(APCI)Calcd.For C 23 H 17 BN2S2[M+H] + :397.0996,found 397.0997.

[0062] Single crystal diffraction pattern Figure 1 shown.

[0063] Example 5

[0064] Preparation of the fluoroboron fluorescent dye derivative represented by formula C5:

[0065]

[0066] The reaction was carried out in the same manner as in Example 3, except that the fluoroboron dipyrrole represented by formula (VIII) was replaced by B2 (54 mg, 0.2 mmol). Post-reaction treatment was the same as in Example 3. Using dichloromethane and petroleum ether as the mobile phase, red solid C5 was obtained by column chromatography (yield 77%).

[0067] The characterization data are as follows: 1 H NMR (500MHz, CDCl3, ppm): δ7.77 (d, J=1.5Hz, 2H), 7.65–7.60 (m, 2H), 7.55–7.51 (m, 5H), 6.96 (dd, J=4.5,1.5Hz,2H),6.53(dd,J=4.5,2.0Hz,2H),6.30(dd,J=3.0,2.0Hz,2H),6.17(d,J=3.0Hz,2H). 13 C NMR (125MHz, CDCl3, ppm): δ146.7,145.4,143.9,134.4,134.0,130.5,130.3,129.5,128.2,117.8,113.7,109.2.HRMS(APCI)Calcd.For C 19H 14 BN2O + [M-C4H3O(furan ring)] + :297.1199,found 297.1200.

[0068] Example 6

[0069] Preparation of the fluoroboron fluorescent dye derivative represented by formula C6:

[0070]

[0071] The reaction was carried out in the same manner as in Example 2, except that the fluoroboron dipyrrole represented by formula (VIII) was replaced by B3 (67 mg, 0.2 mmol). Post-reaction treatment was the same as in Example 2. Using dichloromethane and petroleum ether as the mobile phase, yellow solid C6 (79 mg, 85%) was obtained by column chromatography.

[0072] The characterization data are as follows: 1 H NMR (500MHz, CDCl3, ppm): δ7.76 (s, 2H), 7.49 (d, J = 8.0Hz, 2H), 7.45–7.31 (m, 3H), 7.06 (dd, J=5.0,2.5Hz,2H),7.00(d,J=2.5Hz,2H),6.73(d,J=1.0Hz,2H),6.50(dd,J=4.5,2.5Hz,2H). 13 C NMR (125MHz, CDCl3, ppm): δ146.6,140.2,135.3,133.8,132.0,131.0,130.7,128.1,128.0,127.4,127.1,118.5.HRMS(APCI)Calcd.ForC 23 H 15 BCl2N2S2[M+H] + :465.0217,found 465.0213.

[0073] Single crystal diffraction pattern Figure 1 shown.

[0074] Example 7

[0075] Preparation of fluoroboron fluorescent dye derivatives represented by formula C7:

[0076]

[0077] The reaction was carried out in the same manner as in Example 3, except that the fluoroboron dipyrrole represented by formula (VIII) was replaced by B3 (67 mg, 0.2 mmol). Post-reaction treatment was the same as in Example 3. Using dichloromethane and petroleum ether as the mobile phase, column chromatography was performed to obtain C7 (73 mg, 82%) as a red solid.

[0078] The characterization data are as follows: 1 H NMR (500MHz, CDCl3, ppm): δ7.78 (s, 2H), 7.52–7.46 (m, 4H), 7.40 (dd, J = 9.0, 7.0Hz, 1H), 6.71 (dd, J=4.0,1.0Hz,2H),6.50(dd,J=4.0,1.5Hz,2H),6.30(dd,J=3.0,1.5Hz,2H),6.20(d,J=3.0Hz,2H). 13 C NMR (125MHz, CDCl3, ppm): δ146.3,143.9,140.2,135.3,133.7,132.1,130.9,128.1,127.7,118.4,113.9,109.2.HRMS(APCI)Calcd.For C 19 H 12 BCl2N2O + [M-C4H3O(furan ring)] + :365.0414, found 365.0417. Single crystal diffraction pattern Figure 1 shown.

[0079] Example 8

[0080] Preparation of the fluoroboron fluorescent dye derivative represented by formula C8:

[0081]

[0082] The reaction was carried out according to the method of Example 3, except that the fluoroboron dipyrrole represented by formula (VIII) was replaced by B4 (63 mg, 0.2 mmol). Post-reaction treatment was the same as in Example 3. Dichloromethane and petroleum ether were used as mobile phases, and column chromatography was used to obtain C8, a red solid (yield 81%).

[0083] The characterization data are as follows: 1H NMR (500MHz, CDCl3, ppm): δ8.39(d,J=8.5Hz,2H),7.80(d,J=9.0Hz,4H),7.51(d,J=1.0Hz,2H),6.8 5(d,J=4.5Hz,2H), 6.57(dd,J=4.0,1.0Hz,2H), 6.31(dd,J=3.0,1.5Hz,2H), 6.19(d,J=3.0Hz,2H). 13 C NMR (125MHz, CDCl3, ppm): δ148.9,146.7,144.1,143.2,140.6,133.5,131.3,129.1,123.5,118.7,114.0,109.3.HRMS(APCI)Calcd.For C 19 H 13 BN3O3 + [M-C4H3O(furan ring)] + :342.1044,found 342.1048.

[0084] Single crystal diffraction pattern Figure 1 shown.

[0085] Example 9

[0086] Preparation of the fluoroboron fluorescent dye derivative represented by formula C9:

[0087]

[0088] The reaction was carried out according to the method of Example 2, except that the fluoroboron dipyrrole represented by formula (VIII) was replaced by B5 (41 mg, 0.2 mmol). Post-reaction treatment was the same as in Example 2. Dichloromethane and petroleum ether were used as mobile phases, and column chromatography was used to obtain C9, a red solid (yield 72%).

[0089] The characterization data are as follows: 1 H NMR (500MHz, CDCl3, ppm): δ7.66 (s, 2H) 7.34 (d, J = 3.5Hz, 4H), 7.02 (m, 2H), 6.86 (d, J = 2.5Hz, 2H), 6.53 (dd, J = 3.0, 1.5Hz, 2H), 2.66 (t, J = 4.5Hz, 3H). 13 C NMR (125MHz, CDCl3, ppm): δ145.0,144.7,134.8,130.5,127.3,127.0,126.3,117.3,16.3.HRMS(APCI)Calcd.For C 18 H 15BN2S2[M+H] + :335.0840,found 335.0843.

[0090] Single crystal diffraction pattern Figure 1 shown.

[0091] Test Example 1

[0092] The spectral properties of C1-C9 in dichloromethane were tested, and the test results are shown in Table 1:

[0093] Table 1

[0094] dyes <![CDATA[λ abs max (nm)]]> <![CDATA[ε abs a ]]> <![CDATA[λ em max (nm)]]> <![CDATA[Ф b ]]> <![CDATA[Stokes shift(cm -1 ) c ]]> C1 498 516 4.86 0.44 700 C2 500 515 4.84 0.49 580 C3 497 515 5.02 0.03 700 C4 500 521 4.78 0.02 810 C5 497 518 4.72 <0.01 820 C6 511 528 4.83 0.05 630 C7 507 530 4.73 <0.01 860 C8 505 523 4.65 <0.01 680 C9 494 506 4.96 0.47 480

[0095] In Table 1: Stokes-shift = 1 / λ max –1 / λ em max (cm -1 ), ε max is the molar absorptivity; b Φ is the fluorescence quantum yield.

[0096] It can be seen from the above Examples C1-C9 that the fluoroboron fluorescent dye derivative provided by the present invention and the fluoroboron fluorescent dye derivative prepared by the method provided by the invention have a maximum fluorescence emission wavelength between 494-505 nm, and also have excellent fluorescence quantum yield and excellent solubility, indicating that it has good application prospects in the field of fluorescent labeling. At the same time, the preparation method is simple in steps and the raw materials are easily available.

[0097] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the scope of protection of the present invention.

[0098] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any appropriate manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

[0099] In addition, the various embodiments of the present invention may be arbitrarily combined, and as long as they do not violate the concept of the present invention, they should also be regarded as the contents disclosed by the present invention.

Claims

1. Use of a fluoroboron fluorescent dye derivative in the preparation of a fluorescent marker, characterized in that: The structure of the fluoroboron fluorescent dye derivative is shown in Formula C1-C2. 。 2. The use according to claim 1, characterized in that The preparation method of the fluoroboron fluorescent dye derivative comprises: in the presence of a catalyst, contacting the fluoroboron fluorescent dye of the structure represented by formula (VIII) and a 2-(tributyltinyl) reagent in an organic solution to react, ， In formula (VIII), R2 is 2,4,6-trimethylphenyl; The molar ratio of the compound represented by formula (VIII), the catalyst, and the 2-(tributyltinyl) compound is 0.2:0.2-0.5:0.3-0.5; The catalyst is aluminum chloride; The organic solvent is selected from dichloromethane or toluene; The conditions of the contact reaction include: reaction temperature of 20-30° C., and reaction time of 1-3 h.

3. The use according to claim 2, wherein: The molar ratio of the compound represented by formula (VIII), the catalyst, and the 2-(tributyltinyl) compound is 0.2:0.3-0.4:0.4-0.5.

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