N2O-type BODIPY derivatives, their preparation methods, and applications

Abstract

This invention discloses an N2O-type BODIPY derivative, its preparation method, and its applications. The structure of the N2O-type BODIPY derivative is shown in the following formula, wherein Ar1 is an aryl group; R1 is an aryl group or an aryl derivative; and R2 is selected from one of aryl, aryl derivatives, and nitrogen-aryl groups. This N2O-type BODIPY derivative constructs a new conjugated structure, enabling the boron atom on the original BODIPY molecule to achieve O-B-N six-membered ring closure. This structure promotes the redshift of BODIPY absorption and emission. At the same time, this N2O-type BODIPY derivative has a moderate fluorescence quantum yield (0.07-0.36), and has potential applications in near-infrared cell imaging and other fields. Furthermore, its preparation method has the advantages of mild conditions and simple operation.

Description

Technical Field

[0001] This invention relates to the field of organic synthesis technology, specifically to an N2O type BODIPY derivative, its preparation method, and its application. Background Technology

[0002] BODIPY is one of the most widely used and popular fluorescent dyes, possessing numerous excellent properties such as high fluorescence quantum yield, large molar absorption coefficient, strong absorption and emission bands, and good photostability. It has found increasingly widespread applications in fluorescence imaging and sensing, photodynamic therapy, and optical devices, attracting growing attention from researchers. Because BODIPY fluorophores have abundant functionalization sites, structural modification can enable them to achieve maximum absorption and emission in the near-infrared (NIR) region, thus facilitating their application in near-infrared cell imaging. Summary of the Invention

[0003] The purpose of this invention is to provide an N2O type BODIPY derivative, its preparation method, and its application. This N2O type BODIPY derivative has a novel structure, the raw materials for synthesis are inexpensive and readily available, the synthesis method is simple and efficient, and the preparation method has the advantages of mild conditions and simple operation.

[0004] To achieve the above objectives, the present invention provides an N₂O type BODIPY derivative, the structure of which is shown in the following formula.

[0005]

[0006] Wherein, Ar1 is aryl; R1 is aryl or aryl derivative; R2 is selected from aryl, aryl derivative and nitrogen aryl.

[0007] This invention also provides a method for preparing an N2O type BODIPY derivative, the method comprising:

[0008] (1) The substrate shown in formula (B), acetophenone derivatives and potassium tert-butoxide are reacted in the presence of a solvent to obtain an intermediate product;

[0009] (2) Add boron trifluoride diethyl ether to the intermediate product described in step (1) for coordination reaction, extraction, and column chromatography separation;

[0010]

[0011] Ar1 is an aryl group; R1 is an aryl group or an aryl derivative.

[0012] The present invention also provides an N2O type BODIPY derivative prepared by the above preparation method.

[0013] The present invention further provides an application of the above-mentioned N2O type BODIPY derivative in bioimaging.

[0014] In the above technical solution, the crystal configuration of the N2O type BODIPY derivative of the present invention was obtained by X-ray single crystal diffraction. The N2O type BODIPY derivative of the present invention constructs a new conjugated structure, which enables the boron atoms on the original BODIPY molecule to achieve OBN six-membered ring closure. This structure promotes the redshift of BODIPY absorption and emission.

[0015] Meanwhile, the N2O type BODIPY derivatives of the present invention have good solubility in organic solvents, strong visible light absorption, and large Stokes shift. Compared with BODIPY, their conjugated system is better extended, resulting in a significant redshift in maximum absorption and emission. The maximum absorption and emission wavelengths of some N2O type BODIPY derivatives can reach the near-infrared region.

[0016] Furthermore, the N2O-type BODIPY derivative of this invention has a moderate fluorescence quantum yield (0.07-0.36), and has potential applications in fields such as near-infrared cell imaging.

[0017] Other features and advantages of the present invention will be described in detail in the following detailed description section. Attached Figure Description

[0018] The accompanying drawings are provided to further illustrate the invention and form part of the specification. They are used together with the following detailed description to explain the invention, but do not constitute a limitation thereof. In the drawings:

[0019] Figure 1 These are the crystal structure diagrams of compounds b(a), d(b), f(c), and g(d), respectively.

[0020] Figure 2 These are the molecular structural formulas of product 4a prepared in Example 1, product 4b prepared in Example 2, product 4bb prepared in Example 3, product 4d prepared in Example 5, and product 4f prepared in Example 7, respectively.

[0021] Figure 3 The product 4a prepared in Example 1 exhibits a mirror-image arrangement in its crystal structure;

[0022] Figure 4 The product 4b prepared in Example 2 exhibits a mirror-image arrangement in its crystal structure;

[0023] Figure 5 The product 4bb prepared in Example 3 exhibits a mirror-image arrangement in its crystal structure;

[0024] Figure 6 The product prepared in Example 5 exhibits a mirror-image arrangement in its crystal structure.

[0025] Figure 7 The product 4f prepared in Example 7 exhibits a mirror-image arrangement in its crystal structure;

[0026] Figure 8 This describes the packing pattern of product 4b prepared in Example 2 with adjacent molecules in its crystal structure.

[0027] Figure 9 This describes the packing pattern of product 4d prepared in Example 5 with adjacent molecules in its crystal structure.

[0028] Figure 10 This is the normalized absorption and emission spectrum of test 4a in different solvents in Example 2;

[0029] Figure 11 This is the normalized absorption and emission spectrum of test 4b in different solvents in Example 2;

[0030] Figure 12 This is the normalized absorption and emission spectrum of test 4bb in different solvents in Example 2. Detailed Implementation

[0031] The following provides a detailed description of specific embodiments of the present invention. It should be understood that the specific embodiments described herein are for illustrative and explanatory purposes only and are not intended to limit the scope of the invention.

[0032] The endpoints and any values ​​of the ranges 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 endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0033] This invention provides an N₂O type BODIPY derivative, the structure of which is shown in the following formula.

[0034]

[0035] Wherein, Ar1 is aryl; R1 is aryl or aryl derivative; R2 is selected from aryl, aryl derivative and nitrogen aryl.

[0036] The N2O-type BODIPY derivative of the present invention constructs a new conjugated structure, enabling the boron atoms on the original BODIPY molecule to achieve OBN six-membered ring closure. This structure promotes the redshift of BODIPY absorption and emission.

[0037] In a preferred embodiment of the present invention, Ar1 is phenyl, R1 is selected from phenyl or naphthylcycloyl, and R2 is selected from one of monocyclic aryl, fused aryl and pyridyl.

[0038] In a preferred embodiment of the present invention, Ar1 is selected from mestrimethylphenyl or phenyl, R1 is selected from phenyl, p-methoxyphenyl and 1-naphthyl, and R2 is selected from phenyl, p-morpholinephenyl, 6-methoxy-2-naphthyl, 1-indenyl, 5,6-dimethoxyindenyl, 1-tetrahydronaphthyl, 1-acenaphthyl or 4-pyridyl.

[0039] In a preferred embodiment of the present invention, the structure of the N2O type BODIPY derivative is shown in formula ak.

[0040]

[0041] This invention also provides a method for preparing an N2O type BODIPY derivative, the method comprising:

[0042] (1) The substrate shown in formula (B), acetophenone derivatives and potassium tert-butoxide are reacted in the presence of a solvent to obtain an intermediate product;

[0043] (2) Add boron trifluoride diethyl ether to the intermediate product described in step (1) for coordination reaction, extraction, and column chromatography separation;

[0044]

[0045] Ar1 is an aryl group; R1 is an aryl group or an aryl derivative.

[0046] This invention provides a simple preparation method for obtaining N2O-type BODIPY derivatives. This preparation method is simple to operate and operates under mild conditions.

[0047] In a preferred embodiment of the present invention, Ar1 is a monocyclic aryl group, R1 is selected from monocyclic aryl or fused-ring aryl groups, and the acetophenone derivative is selected from one of acetophenone, 4-morpholinoacetophenone, 6-methoxy-2-acetnaphthalene, 1-indanone, 5,6-dimethoxyindanone, 1-tetrahydronaphthone, 1-acenaphthone, and 4-acetylpyridine.

[0048] In a preferred embodiment of the present invention, Ar1 is selected from mestrimethylphenyl or phenyl, and R1 is selected from phenyl, p-methoxyphenyl or 1-naphthyl.

[0049] In a preferred embodiment of the present invention, in order to obtain more intermediate products, in step (1), the molar ratio of the substrate, acetophenone derivative and potassium tert-butoxide shown in formula (B) is 1:2-4:1-3.

[0050] In a preferred embodiment of the present invention, in step (1), the solvent may be a conventional organic solvent in the art, for example, the solvent may be selected from one or more of acetonitrile, N,N-dimethylformamide and tetrahydrofuran.

[0051] In a preferred embodiment of the present invention, in step (1), the reaction conditions include: a temperature of 20-30°C and a reaction time of 1-3 hours.

[0052] In a preferred embodiment of the present invention, in step (2), the molar ratio of the intermediate product to the boron trifluoride ethyl ether is 1:80-100.

[0053] In a preferred embodiment of the present invention, the conditions for the coordination reaction include: a temperature of 20-30°C and a reaction time of 2-4 hours.

[0054] The present invention also provides an N2O type BODIPY derivative prepared by the above preparation method.

[0055] The present invention further provides an application of the above-mentioned N2O type BODIPY derivative in bioimaging.

[0056] The present invention will be described in detail below through examples, but the scope of protection of the present invention is not limited thereto. In the following examples, the drugs and pharmaceuticals are all conventional commercially available products.

[0057] Preparation Example 1

[0058] Synthesis of substrate B1:

[0059]

[0060] The high-pressure reaction flask of Xinweier was placed in an ice-water bath. 53 mg of the compound shown in formula A1 (0.2 mmol) was dissolved in 8 mL of CH3CN. 50 mg of 2,3-dichloro-5,6-dicyanobenzoquinone (DDQ) (0.22 mmol, 1.1 equiv) was added in portions. The reaction mixture was stirred at this temperature for 30 minutes. Then, 21 mg of anhydrous Na2CO3 (0.2 mmol, 1.0 equiv), 73 mg of PhB(OH)3 (0.6 mmol, 3.0 equiv), and EtOH (1.5 mL, exc) were quickly added to the reaction mixture. The reaction mixture was further stirred in the high-pressure reaction flask at 115 °C for 10 h. After the reaction was completed, the reaction mixture was poured into 30 mL of water and extracted with CH2Cl2 (3 × 30 mL). The organic layers were combined, dried over anhydrous CaCl2, and the solvent was evaporated to dryness using a rotary evaporator. Column chromatography (using ethyl acetate and petroleum ether as eluents) was used to separate the yellow solid B1 (65 mg, 83%).

[0061] 1 H NMR (500MHz, CDCl) 3, ppm): δ7.67(s,2H),7.39(d,J=6.5Hz,2H),7.25-7.15(m,3H),6.98(s,2H),6.65(d,J=3.5Hz,2H),6.41(d d,J=4.0,1.5Hz,2H),3.26(q,J=7.0Hz,2H),2.38(s,3H),2.15(s,3H),2.13(s,3H),1.14(t,J=7.0Hz,3H). 13 C NMR (125MHz, CDCl) 3, ppm): δ147.2,145.3,138.7,136.5,136.3,135.6,131.3,130.8,128.2(9),128 .2(6),127.6,126.7,118.2,57.6,21.4,20.1,20.0,18.3.HRMS(ESI)Calcd.For C 24 H 22 BN2 + [M-OEt] + :349.1871,found 349.1876.

[0062] Preparation Example 2

[0063] Synthesis of substrate B2:

[0064] Following the method of Preparation Example 1, except that PhB(OH)3 was replaced with an equimolar amount of 4-methoxyphenylboronic acid, the reaction yielded substrate B2.

[0065]

[0066] Preparation Example 3

[0067] Synthesis of substrate B3:

[0068] Following the method of Preparation Example 1, except that PhB(OH)3 was replaced with an equimolar amount of 2-naphthoboric acid, the reaction yielded substrate B3.

[0069]

[0070] Preparation Example 4

[0071] Synthesis of substrate B4:

[0072] Following the method of Preparation Example 1, except that the compound shown in Formula A1 was replaced with an equimolar amount of the compound shown in Formula A2, the reaction yielded substrate B4.

[0073]

[0074] Example 1

[0075] Synthetic method of compound a:

[0076] In a 100 mL round-bottom flask, substrate B1 (79 mg, 0.2 mmol), potassium tert-butoxide (45 mg, 0.4 mmol), and acetophenone (51 μL, 0.6 mmol) were added, followed by 20 mL of acetonitrile as solvent. The mixture was stirred at room temperature for approximately 2 hours, and the reaction was monitored by TLC. After substrate B1 had reacted completely, boron trifluoride diethyl ether (4.7 mL, 18 mmol) was added dropwise to the reaction system. The mixture was stirred at room temperature for another 3 hours. The reaction mixture was then poured into a separatory funnel containing 100 mL of saturated saline solution and extracted three times with 30 mL of CH2Cl2. All organic phases were collected and dried over anhydrous Na2SO4. The organic solvent was removed by vacuum distillation. The crude product was purified by column chromatography (300-400 mesh silica gel, petroleum ether / dichloromethane = 4 / 1) to obtain the target product as a red solid a (62 mg, 66%).

[0077]

[0078] The characterization data for compound a are as follows: 1H NMR (500MHz, CDCl3) δ8.07-8.05(m,2H),7.66(s,1H),7.48-7.46(m,3H),7.30(d,J=6.5Hz,2H),7.20-7.11(m,3H),6.97 (d,J=11.5Hz,2H),6.74(d,J=4.5Hz,1H),6.45-6.38(m,3H),6.36-6.31(m,1H),2.37(s,3H),2.28(s,3H),2.09(s,3H). 13 C NMR (125MHz, CDCl3) δ162.9,153.8,138.1,138.0,137.1,136.8,136.1,135.5,134.1,130.5,130 .4,129.9,128.5,128.1,127.4,126.8,126.7,122.4,118.9,115.2,93.6,21.1,20.3,19.9.HRMS calcd.For C 32 H 28 BN2O[M+H] + :467.2289, found 467.2275.

[0079] Example 2

[0080] Synthesis method of compound b:

[0081] The method was followed as in Example 1, except that substrate B1 (79 mg, 0.2 mmol) and 4-morpholinoacetophenone (123 mg, 0.6 mmol) were reacted, and the resulting product was purified by column chromatography to obtain the target product as red solid b (65 mg, 53%).

[0082]

[0083] The characterization data for compound b are as follows: 1 H NMR (500MHz, CDCl3) δ8.00(d,J=9.0Hz,2H),7.61-7.59(m,1H),7.30-7.27(m,2H),7.17-7.11(m,3H),7.98-7.94(m,4H),6.71(d,J=4.5Hz,1H),6. 39(d,J=4.5Hz,1H),6.36(dd,J=4.0,1.5Hz,1H),6.33-6.30(m,2H),3.93 -3.84(m,4H),3.35-3.27(m,4H),2.37(s,3H),2.28(s,3H),2.09(s,3H). 13C NMR (125MHz, CDCl3) δ163.7,154.4,152.7,138.0,137.2,136.9,136.2,134.9,134.3,130.6,130.5,129.9 ,128.4,128.1,127.3,126.7,126.2,121.2,119.3,114.7,114.2,91.8,66.7,48.1,21.1,20.3,19.9.HRMS calcd.For C 36 H 35 BN3O2[M+H] + :552.2817,found 552.2824.

[0084] Example 3

[0085] Synthesis method of compound c:

[0086] The method was the same as in Example 1, except that after reacting substrate B1 (79 mg, 0.2 mmol) and 6-methoxy-2-acetnaphthalene (120 mg, 0.6 mmol), the target product was obtained by column chromatography as a red solid c (65 mg, 67%).

[0087]

[0088] The characterization data for compound c are as follows: 1 H NMR(500MHz, CDCl3)δ8.59(s,1H),8.02(dd,J=8.5,1.5Hz,1H),7.89(d,J=9.0Hz,1H),7 .79(d,J=8.5Hz,1H),7.73(s,1H),7.38-7.31(m,2H),7.22(dd,J=8.5,2.5Hz,1H),7.19 -7.11(m,4H),6.98(d,J=12.5Hz,2H),6.76(d,J=5.0Hz,1H),6.54(s,1H),6.44(dd,J=7 .5,4.5Hz,2H),6.38-6.36(m,1H),3.96(s,3H),2.39(s,3H),2.31(s,3H),2.11(s,3H). 13C NMR (125MHz, CDCl3) δ163.2,158.9,154.0,138.1,137.5,137.1,136.8,136.0,135.9,135.1,134.2,130.7,130.6,130.5(8),130.5(0),1 29.9,128.6,128.1,127.4,126.9,126.8(6),126.8(0),124.2,122.1,119.3,119.1,115.1,105.8,93.5,55.4,21.14,20.3,19.9(8).HRMS calcd.For C 37 H 32 BN2O2[M+H] + :547.2551,found 547.2531.

[0089] Example 4

[0090] Synthesis method of compound d:

[0091] The method was the same as in Example 1, except that after reacting substrate B1 (79 mg, 0.2 mmol) with 1-indanone (79 mg, 0.6 mmol), the target product was obtained by column chromatography as a red solid d (41 mg, 67%).

[0092]

[0093] The characterization data for compound d are as follows: 1 H NMR (500MHz, CDCl3) δ7.90-7.86 (m, 1H), 7.66 (dd, J = 2.0, 1.0Hz, 1H), 7.49-7.42 (m, 2H), 7.40 -7.37(m,1H),7.35-7.29(m,2H),7.16-7.08(m,3H),6.97(d,J=13.0Hz,2H),6.79(d,J=4.5Hz,1H),6.48(d,J=4.5Hz,1 H),6.40(dd,J=3.5,1.0Hz,1H),6.34(dd,J=4.0,2.0Hz,1H),3.67-3.56(m,2H),2.38(s,3H),2.29(s,3H),2.08(s,3H). 13C NMR (125MHz, CDCl3) δ165.9(6),152.1,145.3,138.3,138.1,137.2,136.9,136.8,135.7,134.8,130.6,130.5( 7),128.8,128.1,127.3,127.1,126.7,124.6,121.8,120.7,117.1,115.2,109.8,31.6,21.2,20.3,19.9.HRMS calcd.For C 33 H 28 BN2O[M+H] + :479.2289,found 479.2275.

[0094] Example 5

[0095] Synthetic method of compound e:

[0096] The method was the same as in Example 1, except that after reacting substrate B1 (79 mg, 0.2 mmol) with 5,6-dimethoxyindanone (115 mg, 0.6 mmol), the target product was obtained by column chromatography as a red solid e (52 mg, 48%).

[0097]

[0098] The characterization data for compound e are as follows: 1 H NMR (500MHz, CDCl3) δ7.62(s,1H),7.38(s,1H),7.31(d,J=6.0Hz,2H),7.17-7.09(m,3H),7.04(s,1H),6.97(d,J=13.0Hz,2H),6.77(d,J=4.5Hz ,1H),6.46(d,J=4.5Hz,1H),6.39-6.29(m,2H),4.04(s,3H),3.96(s,3H ),3.56(dd,J=32.5,21.0Hz,2H),2.37(s,3H),2.29(s,3H),2.09(s,3H). 13C NMR (125MHz, CDCl3) δ167.2,152.0,151.1,149.2,139.6,137.9,137.3,137.0,135.02,134.9,134.8,134.5,130.7(7),130.7(1),1 30.6,128.1,127.3,126.7,120.6,117.4,114.7,108.8,107.9,103.1(6),103.1(1),56.2(6),56.2(0),31.4,21.1,20.3,19.9.HRMS calcd.For C 35 H 32 BN2O3[M+H] + :539.2500,found 539.2518.

[0099] Example 6

[0100] Synthetic method of compound f:

[0101] The method of Example 1 was followed, except that after reacting substrate B1 (79 mg, 0.2 mmol) and 1-tetrahydronaphthone (88 mg, 0.6 mmol), the target product was obtained by column chromatography separation and purification as a red solid f (75 mg, 48%).

[0102]

[0103] The characterization data for compound f are as follows: 1 H NMR (500MHz, CDCl3) δ8.16 (dd, J=7.5, 1.5Hz, 1H), 7.65 (dd, J=2.0, 1.0Hz, 1H), 7.40-7.29 (m, 4H), 7.23 -7.02(m,1H),7.18-7.11(m,3H),6.98(d,J=12.0Hz,2H),6.75(d,J=4.5Hz,1H),6.49(d,J=4.5Hz,1H),6.44-6.40(m,1H),6.35(dd,J=4.0 2.0Hz,1H),2.97-2.88(m,2H),2.70-2.65(m,2H),2.39(s,3H),2.31(s,3H). 13CNMR(126MHz, CDCl3)δ156.9,153.7,139.4,138.1,137.7,137.1,136.9,136.2,135.1,134.6,130.6,129.7(8),129.7(5),12 8.1(1),128.1(0),127.3(4),127.2(8),126.8,126.6,124.6,122.2,117.0,115.3,104.6,27.9,22.0,21.1,20.3,19.9.HRMS calcd.ForC 34 H 30 BN2O[M+H] + :493.2446,found 493.2448.

[0104] Example 7

[0105] Synthesis method of compound g:

[0106] The method was the same as in Example 1, except that after reacting substrate B1 (79 mg, 0.2 mmol) and 1-acenaphthene (101 mg, 0.6 mmol), the target product was obtained by column chromatography as a red solid g (84 mg, 82%).

[0107]

[0108] The characterization data for compound g are as follows: 1 H NMR (500MHz, CDCl3) δ8.16(d,J=7.0Hz,1H),7.97(d,J=8.0Hz,1H),7.75(s,1H),7. 72-7.66(m,2H),7.55(d,J=6.5Hz,1H),7.50-7.47(m,1H),7.38(d,J=6.5Hz,2H),7. 18-7.10(m,3H),7.01(d,J=14.5Hz,2H),6.89(d,J=4.5Hz,1H),6.81(d,J=4.5Hz,1H ),6.47(d,J=3.5Hz,1H),6.42-6.38(m,1H),2.40(s,3H),2.32(s,3H),2.12(s,3H). 13CNMR(125MHz, CDCl3)δ168.2,150.4,138.1,137.3,137.2,136.9,136.0,135.0,134.6,133.9,132.4,130.9,130.8(6),130.7,130 .5,129.1,128.4,128.1,127.8,127.3,126.9,124.7,123.5,122.1,119.6,117.4,115.4,109.8,21.2,20.3,19.9.HRMScalcd.For C 36 H 28 BN2O[M+H] + :515.2289,found 515.2308.

[0109] Example 8

[0110] Synthetic method of compound h:

[0111] The method was the same as in Example 1, except that after reacting substrate B2 (85 mg, 0.2 mmol) and acetophenone (51 μL, 0.6 mmol), the target product was obtained by column chromatography as a red solid h (62 mg, 63%).

[0112]

[0113] The characterization data for compound h are as follows: 1 H NMR (500MHz, CDCl3) δ8.09-8.03(m,2H),7.64(dd,J=2.0,1.0Hz,1H),7.49-7.44(m,3H),7.23-7.17(m,2H),6.97(d,J=11 .5Hz,2H),6.76-6.68(m,3H),6.44-6.38(m,3H),6.35-6.30(m,1H),3.70(s,3H),2.37(s,3H),2.27(s,3H),2.09(s,3H). 13 C NMR (125MHz, CDCl3) δ163.0,158.8,153.8,138.1,137.9,137.1,136.8,136.2,135.6,135.2,134.1,131.7 ,130.5,130.4,129.8,128.5,128.1,126.7,122.3,118.9,115.2,113.0,93.5,54.9,21.1,20.3,19.9.HRMS calcd.For C 33 H 30BN2O2[M+H] + :497.2395,found497.2391.

[0114] Example 9

[0115] Synthetic method of compound i:

[0116] The method was the same as in Example 1, except that after reacting substrate B3 (89 mg, 0.2 mmol) and acetophenone (51 μL, 0.6 mmol), the target product was obtained by column chromatography as a red solid i (56 mg, 54%).

[0117]

[0118] The characterization data for compound i are as follows: 1 H NMR (500MHz, CDCl3) δ8.12-8.08(m,2H),7.72-7.68(m,3H),7.67-7.61(m,2H),7.53(d,J=8.0Hz,1H),7.51-7.47(m,3H),7.35-7.31 (m,2H),6.99(d,J=10.0Hz,2H),6.79(d,J=4.5Hz,1H),6.46-6.42(m,3H),6.35-6.33(m,1H),2.39(s,3H),2.34(s,3H),2.12(s,3H). 13 CNMR(125MHz, CDCl3)δ163.0,153.9,138.2,138.1(7),137.1,136.8,136.3,135.6,135.2,134.2,133.2,133.0,130.5,130. 0,129.6,128.8,128.5,128.2,127.9,127.4,126.7,126.6,125.0,124.8,122.6,119.0,115.3,93.7,21.1,20.3,19.9.HRMS calcd.For C 36 H 30 BN2O[M+H] + :517.2446,found 517.2437.

[0119] Example 10

[0120] Synthetic method of compound j:

[0121] The method was the same as in Example 1, except that after reacting substrate B3 (71 mg, 0.2 mmol) and acetophenone (51 μL, 0.6 mmol), the target product was obtained by column chromatography as a red solid j (51 mg, 60%).

[0122]

[0123] The characterization data for compound j are as follows: 1 H NMR (500MHz, CDCl3) δ8.09 -8.01(m,2H),7.71-7.64(m,3H),7.56-7.49(m,3H),7.48-7.44(m,3H),7.28-7.25(m,2H),7.17-7.10(m,3H),6. 96(d,J=5.0Hz,1H),6.81(dd,J=4.0,1.0Hz,1H),6.48(d,J=4.5Hz,1H),6.44(dd,J=4.0,2.0Hz,1H),6.41(s,1H). 13 C NMR (125MHz, CDCl3) δ163.2,158.2,153.9,152.9,138.3,136.3,135.5,134.7,134.6,133.9,131.3,1 30.5(2),130.4(6),130.3,129.8,128.5,128.4,127.4,126.8,126.7,123.5,119.2,115.4,93.5.HRMS calcd.For C 29 H 22 BN2O[M+H] + :425.1820,found 425.1826.

[0124] Example 11

[0125] Synthetic method of compound k:

[0126] The method was the same as in Example 1, except that after reacting substrate B1 (79 mg, 0.2 mmol) and 4-acetylpyridine (73 mg, 0.6 mmol), the target product was obtained by column chromatography as a red solid k (64 mg, 69%).

[0127]

[0128] The characterization data for compound k are as follows: 1H NMR (500MHz, DMSO) δ8.73(d,J=4.5Hz,2H),8.03(d,J=5.0Hz,2H),8.01(s,1H),7.19(d,J=6.5Hz,2H),7.15-7.03(m,5H),6.98(s,1H ),6.87(d,J=4.5Hz,1H),6.69(d,J=4.5Hz,1H),6.43(d,J=2.5Hz,1H),6.37(d,J=4.0Hz,1H),2.34(s,3H),2.17(s,3H),2.01(s,3H). 13 C NMR (125MHz, DMSO) δ158.0,152.1,150.1,141.6,139.2,138.1,137.8,135.8,135.5,134.5,133.3,129 .7,129.6,127.9(9),127.9(5),127.1,126.5,123.2,119.6,119.3,116.2,96.1,20.5,19.4,19.2.HRMS calcd.ForC 31 H 27 BN3O[M+H] + :468.2242,found 468.2244.

[0129] Detection Example 1

[0130] The optical properties of substrate B1 in dichloromethane and the optical properties of products aj prepared in Examples 1-10 in n-hexane, toluene, dichloromethane, tetrahydrofuran, and acetonitrile were tested at room temperature. The results are shown in Table 1 and... Figure 2-11 As shown.

[0131] Table 1. Spectroscopic and photophysical data of B1 in dichloromethane and compounds 5a-5j in different organic solvents at room temperature.

[0132]

[0133]

[0134]

[0135] Depend on Figure 2-11As can be observed from Table 1, compared to the control compound B1, the N2O-type BODIPY derivatives prepared in Examples 1-10 of this invention exhibit a significant red shift in absorption and emission peaks after the boron atom of BODIPY achieves OBN six-membered ring closure. The maximum absorption peak shifts from 500 nm to over 600 nm, and the maximum emission also red-shifts to 650 nm, showing a red shift of over 120 nm, reaching the near-infrared region. This indicates that constructing an OBN six-membered ring closure at the boron site is an effective strategy to promote the red shift of BODIPY absorption and emission. Increasing the electron-donating ability of the phenyl group on the OBN six-membered ring is beneficial to red-shifting the absorption and emission wavelength of BODIPY. For example, in n-hexane solution, the maximum absorption and emission peaks of a are located at 617 and 644 nm, respectively. When an electron-donating morpholino group is installed at the para-position of the phenyl group on the OBN six-membered ring of a, i.e., molecule b, its absorption and emission are red-shifted by 15 nm and 17 nm, respectively. When the phenyl group is replaced with a p-naphthyl group, its absorption and emission are red-shifted to 632 nm and 652 nm, respectively.

[0136] Although α exhibits a significant redshift in both absorption and emission, it still maintains a high fluorescence quantum yield (0.28-0.36) in various organic solvents. Considering that fixing the phenyl group on the OBN six-membered ring of α with a methyl or ethyl group through conformational restriction to the OBN six-membered ring might also achieve a redshift in absorption and emission peaks and improve fluorescence quantum yield, molecules d, e, and f were synthesized. Consistent with expectations, conformational restriction of the phenyl group with a methyl or ethyl group did indeed achieve a redshift in absorption and emission. The absorption and emission peaks of d were 638 nm and 678 nm, those of e were 652 nm and 684 nm, and those of f were 642 nm and 678 nm. However, the fluorescence was not enhanced as expected, especially the fluorescence of e, which was further quenched to 0.07. This may be because the loss of fluorescence ability in α originates from the rotational nonradiative transition loss of the other phenyl group on the boron ring, while the two methoxy substituents on the phenyl group of e's O-BN six-membered ring quenched the molecule's fluorescence due to a strong electron-donating effect.

[0137] To further enhance the conjugation of the OBN six-membered ring phenyl group, resulting in a redshift in the absorption and emission of the product, the substrate acetophenone of compound a was replaced with 1-acenaphthene to synthesize compound g. The maximum absorption and emission wavelengths of g reached 680 nm and 712 nm, respectively, representing redshifts of 63 nm and 68 nm compared to a. In addition, the other phenyl group on the boron of compound a was modified by replacing it with p-methoxyphenyl and naphthyl groups, respectively, yielding compounds h and i. Experiments showed that the modification of the other phenyl group on the boron did not significantly improve the absorption and emission of the compounds; the absorption and emission peaks of h and i in n-hexane were both 616 nm and 642 nm, respectively. Their fluorescence quantum yields were also not significantly different from those of compound a.

[0138] Application Example 1

[0139] Given that the morpholine group contained in product b prepared in Example 2 has potential lysosomal localization ability, product b was selected for further cell imaging studies. The specific steps are as follows:

[0140] Cell culture: HeLa cells were cultured for 24 h in RPMI-1640 medium containing 10% FBS and 1% penicillin / streptomycin in an incubator at 37°C, 5% CO2 and 95% humidity.

[0141] Cellular colocalization: Compound b prepared in Example 2 was used for intracellular lysosomal localization studies.

[0142] HeLa cells were co-incubated for 3 hours with b (100 nM) and the lysosomal dye LysoTracker-blue (as reported in the literature). Images were then captured using confocal fluorescence microscopy (blue channel: LysoTracker-blue fluorescence, excitation at 405 nm; green channel: b fluorescence, excitation at 638 nm). The results are as follows: Figure 12 As shown.

[0143] Before cell imaging, the cytotoxicity of compound b was detected by standard CCK-8 assay, and the results showed that compound b has good biocompatibility.

[0144] Depend on Figure 12 Subsequent colocalization experiments showed that b covered the lysosomal probe well, and the subcellular localization of b was almost identical to that of the lysosomal-specific probe LysoTracker-blue in live HeLa cells. Furthermore, when the region of interest (white line) was crossed out in HeLa cells, the fluorescence intensity distributions of b and LysoTracker-blue were very similar, with Pearson correlation coefficients of 0.80 and 0.83, respectively, indicating that b has the potential for lysosomal-specific imaging.

[0145] Detection Example 2

[0146] Cytotoxicity assay:

[0147] HeLa cells were dispersed in 1640 complete medium (7000 cells / well) and seeded in 96-well plates. After culturing for 24 hours, the medium in the 96-well plates was removed, and fresh medium containing 0-100 μM gradient concentrations of 4bb or 4f was added and cultured for another 24 hours. Then, the working solution was removed, and the cells were carefully washed once with PBS. After removing the PBS, a 10-fold diluted CCK-8 solution (Cell Counting Kit-8, BIOMIKY) was added to each well. The cells were then transferred to an incubator for 1 hour. After shaking the 96-well plates for 5 minutes, the absorbance at 450 nm was read using a microplate reader (Multiskan Sky). Before cell imaging, the cytotoxicity of 4bb and 4f was assessed using standard CCK-8.

[0148] As can be seen from the above, more than 90% of the cells can still survive at a concentration as high as 40 μM, indicating that the BOSPY of the present invention has good biocompatibility.

[0149] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of 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 protection scope of the present invention.

[0150] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

[0151] Furthermore, various different embodiments of the present invention can be combined in any way, as long as they do not violate the spirit of the present invention, they should also be regarded as the content disclosed by the present invention.

Claims

1. An N2O type BODIPY derivative, characterized in that, The structure of the N2O type BODIPY derivative is shown in the following formula. ； Wherein, Ar1 is selected from mestrimethylphenyl or phenyl, R1 is selected from phenyl, p-methoxyphenyl and 1-naphthyl, and R2 is selected from phenyl, p-morpholinephenyl, 6-methoxy-2-naphthyl, 1-indenyl, 5,6-dimethoxyindenyl, 1-tetrahydronaphthyl, 1-acenaphthyl or 4-pyridyl.

2. The N2O type BODIPY derivative according to claim 1, characterized in that, The structure of the N2O type BODIPY derivative is shown in formula ak. 。 3. A method for preparing the N2O type BODIPY derivative as described in claim 1 or 2, characterized in that, The preparation method includes: (1) The substrate shown in formula (B), acetophenone derivatives and potassium tert-butoxide are reacted in the presence of a solvent to obtain an intermediate product; (2) Add boron trifluoride diethyl ether to the intermediate product described in step (1) for coordination reaction, extraction, and column chromatography separation; ； The acetophenone derivative is selected from one of acetophenone, 4-morpholinoacetophenone, 6-methoxy-2-acetnaphthalene, 1-indanone, 5,6-dimethoxyindanone, 1-tetrahydronaphthone, 1-acenaphthene, and 4-acetylpyridine; Ar1 ​​is selected from mestrimethylphenyl or phenyl, and R1 is selected from phenyl, p-methoxyphenyl, or 1-naphthyl.

4. The preparation method according to claim 3, characterized in that, In step (1), the molar ratio of the substrate shown in formula (A), the acetophenone derivative, and potassium tert-butoxide is 1:2-4:1-3; the solvent is selected from one or more of acetonitrile, N,N-dimethylformamide, and tetrahydrofuran.

5. The preparation method according to claim 3, characterized in that, In step (1), the reaction conditions include: a temperature of 20-30°C and a reaction time of 1-3 hours.

6. The preparation method according to claim 3, characterized in that, In step (2), the molar ratio of the intermediate product to the boron trifluoride diethyl ether is 1:80-100; the conditions for the coordination reaction include: a temperature of 20-30℃ and a reaction time of 2-4h.

7. The use of an N2O type BODIPY derivative as described in claim 1 or 2 in the preparation of bioimaging reagents.

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