An Aza-BODIPY-like compound, its preparation method and application

Abstract

This invention discloses an Aza-BODIPY type compound, its preparation method, and its applications. This type of compound has the structure shown in Formula I, wherein R is selected from -(CH2CH2O). n CH3, (CH2) k SO3H, n≥1; k≥1; R1 and R2 are selected from -CH2(OCH2CH2) m CH3; m≥1. This invention provides a novel Aza-BODIPY-type compound, which exhibits strong fluorescence, high water solubility, and high selectivity; it can be used for the detection of carbon monoxide and has a low detection limit.

Description

Technical Field

[0001] This invention relates to the field of fluorescent probe technology, and in particular to an Aza-BODIPY-type compound, its preparation method, and its application. Background Technology

[0002] Carbon monoxide (CO), as an important gaseous signaling molecule, plays a crucial role in various physiological and pathological processes, including neurotransmission and anti-apoptotic sepsis. Abnormal CO metabolism in organisms can lead to diseases such as Alzheimer's disease, hypertension, and heart failure. Furthermore, CO can inhibit cytochrome c oxidase and the mitochondrial respiratory chain, resulting in increased reactive oxygen species (ROS), decreased ATP, damage to mitochondrial function, and apoptosis. In addition, excessive CO in the environment can bind to hemoglobin in the blood, reducing blood oxygenation capacity and causing tissue hypoxia. Therefore, CO detection is of great significance in biomedical research and environmental monitoring. Currently, CO detection methods mainly include electrochemical methods, chromatographic analysis, and colorimetric methods, but these methods suffer from slow detection speed, low sensitivity, complex equipment, high cost, and the inability to perform real-time, non-invasive monitoring of CO in living organisms.

[0003] Therefore, it is urgent to develop a new type of fluorescent probe that has good water solubility, high selectivity, obvious effect, low cost, fast response speed, high sensitivity, and can perform real-time non-invasive monitoring in vivo. Summary of the Invention

[0004] The present invention aims to at least solve one of the technical problems existing in the prior art. To this end, the first aspect of the present invention proposes an Aza-BODIPY type compound, which exhibits good water solubility and high selectivity for the fluorescence detection of carbon monoxide.

[0005] A second aspect of the present invention also provides a method for preparing Aza-BODIPY-type compounds.

[0006] A third aspect of the present invention also provides a fluorescent probe.

[0007] The fourth aspect of the present invention also provides an application of Aza-BODIPY-type compounds.

[0008] The Aza-BODIPY-type compound provided according to the first aspect of the present invention has the structure shown in Formula I:

[0009]

[0010] Wherein, R is selected from -(CH2CH2O). n CH3, (CH2) k SO3H, n≥1;k≥1;

[0011] R1 and R2 are selected from -CH2(OCH2CH2) m CH3; m≥1.

[0012] The Aza-BODIPY compounds according to embodiments of the present invention have at least the following beneficial effects:

[0013] This invention provides a novel Aza-BODIPY-type compound, which exhibits strong fluorescence, high water solubility, and high selectivity; it can be used for the detection of carbon monoxide and has a low detection limit.

[0014] According to some embodiments of the present invention, 1 ≤ n ≤ 20.

[0015] According to some embodiments of the present invention, 1 ≤ m ≤ 20.

[0016] According to some embodiments of the present invention, 1≤k≤20.

[0017] According to some embodiments of the present invention, the Aza-BODIPY-type compounds are selected from one of the following structural formulas:

[0018]

[0019]

[0020] The method for preparing Aza-BODIPY-type compounds according to a second aspect embodiment of the present invention includes the following steps:

[0021] The compound 6, allyl chloroformate, organic base, and organic solvent are mixed and reacted to obtain the product.

[0022] The structural formula of compound 6 is as follows:

[0023]

[0024] According to some embodiments of the present invention, the temperature of the reaction is -5 to 5°C.

[0025] According to some embodiments of the present invention, the temperature of the reaction is 0 to 5°C.

[0026] According to some embodiments of the present invention, the organic base is selected from at least one of DIPEA, DEA, TEA, and piperidine.

[0027] According to some embodiments of the present invention, the organic solvent is selected from at least one of dichloromethane, tetrahydrofuran, dimethyl sulfoxide, N'N-dimethylformamide, acetonitrile, or ethanol.

[0028] According to some embodiments of the present invention, the reaction time is 20 to 80 minutes.

[0029] According to some embodiments of the present invention, compound 6 is prepared by the following method:

[0030] The compound 5, an inorganic base, an organic solvent, and a compound substituted with the R group are mixed and reacted to obtain the product.

[0031] The structural formulas of compound 5 and the compounds substituted with the R group are as follows:

[0032]

[0033] According to some embodiments of the present invention, the inorganic base is selected from at least one of cesium carbonate, potassium carbonate, and sodium hydroxide.

[0034] According to some embodiments of the present invention, the organic solvent is selected from at least one of acetonitrile, ethanol, and tetrahydrofuran.

[0035] According to some embodiments of the present invention, the reaction temperature is 60–100°C.

[0036] According to some embodiments of the present invention, compound 5 is prepared by commercially available methods or according to methods disclosed in the following documents:

[0037] Gorman,A.,Killoran,J.,O'Shea,C.,Kenna,T.,Gallagher,WM,&O'Shea,DFJOURNAL OF THE AMERICAN CHEMICAL SOCIETY 6(2004)10619-10631.

[0038] A third aspect of the present invention provides a fluorescent probe comprising the Aza-BODIPY class compounds described in the first aspect of the present invention.

[0039] Furthermore, since it incorporates all the technical features of the Aza-BODIPY class compounds described in the first aspect of the present invention, the fluorescent probe of the present invention has all the technical effects of the Aza-BODIPY class compounds of the present invention.

[0040] According to some embodiments of the present invention, the concentration of the fluorescent probe in the solvent is 10. -7 ~10 -2 mol / L.

[0041] According to some embodiments of the present invention, the solvent includes DMSO and / or H2O.

[0042] The fourth aspect of this invention provides the use of the aforementioned Aza-BODIPY compounds; or the aforementioned fluorescent probe in the detection of carbon monoxide for non-diagnostic or therapeutic purposes.

[0043] Other features and advantages of the invention will be set forth in the description which follows, and will be apparent in part from the description, or may be learned by practicing the invention. Attached Figure Description

[0044] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the description of the embodiments taken in conjunction with the following drawings, in which:

[0045] Figure 1 This is the 1H NMR spectrum of compound AB1-CO prepared in Example 1 of this invention;

[0046] Figure 2 This is the 1H NMR spectrum of compound AB2-CO prepared in Example 2 of this invention;

[0047] Figure 3 This is a high-resolution mass spectrum of compound AB1-CO prepared in Example 1 of this invention;

[0048] Figure 4 This is a high-resolution mass spectrum of compound AB2-CO prepared in Example 2 of this invention;

[0049] Figure 5 This is a particle size distribution diagram of compound AB1-CO prepared in Example 1 of this invention in an aqueous phase.

[0050] Figure 6 This is a particle size distribution diagram of compound AB2-CO prepared in Example 2 of this invention in the aqueous phase.

[0051] Figure 7 The time-dependent UV absorption spectra of the AB1-CO solution prepared in Example 1 of this invention before and after the reaction with CO are shown.

[0052] Figure 8 The time-dependent UV absorption spectra of the AB2-CO solution prepared in Example 2 of this invention before and after the reaction with CO are shown.

[0053] Figure 9 This is a time-dependent fluorescence intensity change diagram of the AB1-CO solution prepared in Example 1 of the present invention before and after the reaction with CO;

[0054] Figure 10 This is a time-dependent fluorescence intensity change diagram of the AB2-CO solution prepared in Example 2 of the present invention before and after the reaction with CO;

[0055] Figure 11The graph shows the linear relationship between fluorescence intensity and CO concentration in the reaction of AB1-CO prepared in Example 1 of this invention with different concentrations of CO.

[0056] Figure 12 The graph shows the linear relationship between fluorescence intensity and CO concentration in the reaction of AB2-CO prepared in Example 2 of this invention with different concentrations of CO.

[0057] Figure 13 The time-dependent UV absorption spectra of the solutions of AB1-CO prepared in Example 1 of this invention before and after reaction with other analytes are shown.

[0058] Figure 14 The time-dependent UV absorption spectra of the AB2-CO solution prepared in Example 2 of this invention before and after reaction with other analytes are shown. Detailed Implementation

[0059] The following are specific embodiments of the present invention, and the technical solutions of the present invention will be further described in conjunction with the embodiments, but the present invention is not limited to these embodiments.

[0060] Unless otherwise specified, the reagents, methods and equipment used in this invention are all conventional reagents, methods and equipment in this technical field.

[0061] Example 1

[0062] This example provides an Aza-BODIPY-type compound, with the following structural formula and preparation method:

[0063]

[0064] Preparation of compounds substituted with R groups, with the following structural formula:

[0065]

[0066] Polyethylene glycol monomethyl ether (100 mg, 0.83 mmol) and NaOH (499 mg, 12.48 mmol) were weighed into a 100 mL round-bottom flask. THF (10 mL) was added, and the mixture was stirred for 30 min in an ice bath. p-Toluenesulfonyl chloride (238 mg, 1.25 mmol) was dissolved in a small amount of THF and added dropwise to the round-bottom flask in an ice bath. The reaction was monitored by TLC. After the reaction was terminated, dilute hydrochloric acid solution was added to the filtrate, and the mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain 199 mg of a colorless oil, with a yield of 87%. 1H NMR(500MHz,Chloroform-d)δ7.80(d,J=8.3Hz,2H),7.34(d,J=8.1Hz,2H),4.18-4.15(m ,2H),3.71-3.67(m,2H),3.59-3.56(m,2H),3.50-3.46(m,2H),3.35(s,3H),2.44(s,3H).

[0067] Compound 5 was prepared according to the method disclosed in the literature Gorman, A., Killoran, J., O'Shea, C., Kenna, T., Gallagher, WM, & O'Shea, DF JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 6 (2004) 10619-10631.

[0068] Synthesis of compound 6:

[0069] The reaction equation and preparation method are as follows:

[0070]

[0071] Compound 5 (100 mg, 0.19 mmol) and Cs₂CO₃ (154 mg, 47.2 mmol) were weighed into a 100 mL double-necked flask. MeCN (10 mL) was added, the flask was evacuated, purged with nitrogen, and heated to reflux at 80 °C. After stirring for 30 min, the R-substituted compound (51.8 mg, 0.19 mmol) was slowly added dropwise to the flask. The reaction was monitored by TLC. After the reaction was terminated, dilute hydrochloric acid solution (pH = 1) was added to the filtrate, and the mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain 77 mg of a red solid, with a yield of 65%. 1 H NMR (500MHz, DMSO-d6) δ10.59(s,1H),8.22-8.08(m,8H),7.64(s,1H),7.59-7.42(m,7H),7.15(d,J=8.6Hz,2H),6.95(d, J=8.5Hz,2H),4.24(t,J=4.6Hz,2H),3.78(t,J=4.5Hz,2H),3.65-3.57(m,2H),3.48(dd,J=5.8,3.6Hz,2H),3.26(s,3H).

[0072] Synthesis of AB1-CO

[0073] Compound 6 (100 mg, 0.158 mmol) was weighed into a 100 mL flask, and DCM (10 mL) was added. Under ice bath conditions, DIPEA (61.4 mg, 47.5 mmol) and allyl chloroformate (28.6 mg, 0.24 mmol) were added dropwise. The reaction was monitored by TLC. After the reaction was terminated, dilute hydrochloric acid solution at pH=1 was added to the filtrate, and the mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain 101 mg of red solid, with a yield of 89%.

[0074] The obtained product was analyzed by proton nuclear magnetic resonance spectroscopy and high-resolution mass spectrometry, and the results are as follows: Figure 1 and Figure 3 As shown: 1 HNMR(500MHz,Chloroform-d)δ8.13-8.03(m,8H),7.50-7.39(m,6H),7.32(d,J=8.8Hz,2H),7.1 2(d,J=1.1Hz,1H),7.04(d,J=8.9Hz,2H),6.97(d,J=1.2Hz,1H),6.03(ddt,J=17.3,10.5,5.9Hz, 1H),5.46(dd,J=17.2,1.4Hz,1H),5.36(dd,J=10.4,1.2Hz,1H),4.78(dt,J=5.9,1.3Hz,2H),4. 27-4.21(m,2H),3.92-3.87(m,2H),3.77-3.71(m,2H),3.62-3.57(m,2H),3.41(s,3H),HRMS:m / z calcd for[C 41 H 36 BF2N3O6+H] + 715.2774, found: 715.2769.

[0075] Example 2

[0076] This example provides an Aza-BODIPY-type compound, with the following structural formula and preparation method:

[0077]

[0078] The synthesis of compounds substituted with R groups, along with their reaction equations and preparation methods, are as follows:

[0079]

[0080] The specific synthesis steps are as follows:

[0081] (1) Synthesis of Compound 1

[0082] 3-Chloro-2-chloromethylpropene (100 mg, 0.8 mmol) and 60% NaH (96 mg, 2.4 mmol) were weighed into a double-necked flask. THF (10 mL) was added, and the mixture was stirred at 65 °C for 30 min. Triethylene glycol monomethyl ether (263 mg, 1.6 mmol) was dissolved in a small amount of THF and added dropwise to the double-necked flask. The reaction was monitored by TLC. After the reaction was terminated, dilute hydrochloric acid solution (pH = 1) was added to the filtrate, and the mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain 278 mg of a colorless oil, with a yield of 87%. 1 H NMR (500MHz, Chloroform-d) δ 5.19-5.17 (m, 2H), 4.01 (d, J = 1.1Hz, 4H), 3.68-3.62 (m, 16H), 3.56 (ddd, J = 16.2, 5.9, 3.8Hz, 8H), 3.37 (s, 6H).

[0083] (2) Synthesis of compounds with R-group substitution

[0084] Compound 1 (100 mg, 0.263 mmol) was weighed into a 100 mL double-necked flask, THF (10 mL) was added, the flask was evacuated and purged with nitrogen, and 9-BBN (48.1 mg, 0.394 mmol) was added dropwise under ice bath conditions. After stirring for 1 h, NaOH solution (26.3 mg, 0.657 mmol) was added dropwise under ice bath conditions. After stirring for 45 min, H2O2 solution (22.3 mg, 0.657 mmol) was added dropwise. The reaction was monitored by TLC. After the reaction was terminated, dilute hydrochloric acid solution with pH=1 was added to the filtrate, and the mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product compound 2.

[0085] Compound 2 (100 mg, 0.25 mmol) and NaOH (151 mg, 3.8 mmol) were weighed into a 100 mL double-necked flask. THF (10 mL) was added, and p-toluenesulfonyl chloride (71.8 mg, 0.38 mmol) was added dropwise under conditions of T < 5 °C. The mixture was stirred at room temperature, and the reaction was monitored by TLC. After the reaction was terminated, a dilute hydrochloric acid solution with pH = 1 was added to the filtrate, and the mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain 87 mg of a colorless oily substance, with a yield of 63%. 1 H NMR(500MHz,Chloroform-d)δ7.80-7.75(m,2H),7.36-7.32(m,2H),4.11(d,J=5.5Hz,2H),3.66-3. 60(m,12H),3.57-3.52(m,8H),3.50-3.39(m,8H),3.37(s,6H),2.44(s,3H),2.24(p,J=6.0Hz,1H).

[0086] Synthesis of Compound 6

[0087] The reaction equation and preparation method are as follows:

[0088]

[0089] Compound 5 (100 mg, 0.19 mmol) and Cs₂CO₃ (154 mg, 0.47 mmol) were weighed into a 100 mL double-necked flask. 10 mL of MeCN was added, the flask was evacuated, purged with nitrogen, and heated to reflux at 80 °C. After stirring for 30 min, the water-soluble chain R₂ (104 mg, 0.19 mmol) was slowly added dropwise to the flask. The reaction was monitored by TLC. After the reaction was terminated, dilute hydrochloric acid solution (pH = 1) was added to the filtrate, and the mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain 101 mg of a red solid, with a yield of 59%. 1 H NMR(500MHz,Chloroform-d)δ8.05-7.98(m,8H),7.44-7.33(m,6H),7.01(s,1H),6.99-6.90 (m,5H),4.12(dd,J=6.4,4.1Hz,2H),3.67-3.53(m,28H),3.37(s,6H),2.41(p,J=5.9Hz,1H).

[0090] Synthesis of probe AB2-CO

[0091] Compound 6 (100 mg, 0.11 mmol) was weighed into a 100 mL flask, and DCM (10 mL) was added. Under ice bath conditions, DIPEA (42.6 mg, 0.33 mmol) and allyl chloroformate (19.9 mg, 0.16 mmol) were added dropwise. The reaction was monitored by TLC. After the reaction was terminated, dilute hydrochloric acid solution at pH=1 was added to the filtrate, and the mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain 99 mg of red solid, with a yield of 91%.

[0092] The prepared product was analyzed by 1H NMR and high-resolution mass spectrometry, and its 1H NMR spectrum is shown below. Figure 2 As shown, high-resolution mass spectrometry is as follows Figure 4 As shown.

[0093] 1H NMR(500MHz,Chloroform-d)δ8.14-8.02(m,8H),7.50-7.38(m,7H),7.32(d,J=8.5Hz,2H ),7.12(s,1H),7.03(d,J=8.7Hz,2H),6.96(s,1H),6.02(ddt,J=16.5,10.9,5.8Hz,1H), 5.46(d,J=17.2Hz,1H),5.36(d,J=10.4Hz,1H),4.77(d,J=5.9Hz,2H),4.14(d,J=5.5Hz, 2H),3.68–3.58(m,24H),3.56-3.52(m,4H),3.36(s,6H),2.45(p,J=5.9Hz,1H),HRMS:m / z calcd for[C 54 H 62 BF2N3O 12 -H] + 991.4347, found: 991.4347.

[0094] Performance testing

[0095] Water solubility test

[0096] To verify the water solubility of the two compounds prepared in Examples 1 and 2 of this invention, AB1-CO and AB2-CO were dissolved in THF solution to obtain probe stock solution (10 -3 mol / L), take 2 mL of ultrapure water into a sample bottle, add the probe under ultrasonic conditions to obtain 10 -5 A mol / L aqueous solution of AB1-CO and AB2-CO nanoparticles was prepared, followed by heating to remove residual THF, and the solution was filtered through a filter membrane. Dynamic light scattering assays showed that AB1-CO and AB2-CO exhibited good dispersion properties in the aqueous phase, thus demonstrating the high water solubility of the two probes (e.g., mol / L). Figure 5 and Figure 6 Furthermore, the two probes, AB1-CO and AB2-CO, have sizes of 50 nm and 100 nm, respectively. This probe size range indicates good blood circulation capability in live animal experiments, and the probes have great potential for live imaging.

[0097] Fluorescence performance detection

[0098] The AB1-CO and AB2-CO prepared in Examples 1 and 2 of this invention were dissolved in THF solution to obtain probe stock solution (10 - 3 After preparing the test system of DMSO:H2O = 7:3 (mol / L), the probe was added to obtain 10 -5Add 10 mol / L AB1-CO and AB2-CO solutions to PdCl2 solution (10 mol / L). -5 mol / L) and CORM-3 (carbon monoxide releasing molecules) solution (20×10) -3 The UV spectrum of the reaction (mol / L) after heating for 30 minutes was changed from... Figure 7 and Figure 8 It can be seen that when Pd 2+ Reduced to Pd by CO 0 The Tsuji-Trost reaction then occurred, causing a red shift in the wavelengths of AB1-CO and AB2-CO, resulting in a new absorption peak at 695 nm.

[0099] Further, the AB1-CO and AB2-CO prepared in Examples 1 and 2 of this invention were dissolved in THF solution to obtain probe stock solution (10 -3 After preparing the test system of DMSO:H2O = 7:3 (mol / L), the probe was added to obtain 10 -5 Add 10 mol / L AB1-CO solution and AB2-CO solution to PdCl2 solution (10 mol / L) -5 mol / L) and CORM-3 solution (20×10 -3 The fluorescence intensity at 716 nm was measured after heating the reaction (mol / L) for 30 minutes. Figure 9 and Figure 10 It can be seen that when Pd 2+ Reduced to Pd by CO 0 The Tsuji-Trost reaction then occurred, causing a redshift of approximately 25 nm in the maximum emission wavelengths of AB1-CO and AB2-CO, enabling the detection of CO through spectral response.

[0100] Further, the AB1-CO and AB2-CO prepared in Examples 1 and 2 of this invention were dissolved in THF solution to obtain probe stock solution (10 -3 After preparing the test system of DMSO:H2O = 7:3 (mol / L), the probe was added to obtain 10 -5 Add 10 mol / L AB1-CO and AB2-CO solutions to PdCl2 solution (10 mol / L). -5 The fluorescence intensity of CORM-3 solutions at different concentrations (mol / L) was measured, and the changes were observed from... Figure 11 and Figure 12 It can be seen that there is a good linear relationship between the emission peak intensity at 716 nm and the CORM-3 concentration. Based on this linear relationship, the detection limit of the AB1-CO probe is as low as 104 nM, and the detection limit of the AB2-CO probe is as low as 43.4 nM. The detection limits of AB1-CO and AB2-CO are at a low level.

[0101] Selective detection

[0102] The AB1-CO and AB2-CO prepared in Examples 1 and 2 of this invention were dissolved in THF solution to obtain probe stock solution (10 - 3 After preparing the test system of DMSO:H2O = 7:3 (mol / L), the probe was added to obtain 10 -5 Add 10 mol / L AB1-CO and AB2-CO solutions to PdCl2 solution (10 mol / L). -5 The absorbance changes of solutions of other analytes (mol / L) and solutions with a concentration twice that of CORM-3 were measured. Figure 13 and Figure 14 It can be seen that the maximum absorption wavelengths of the analytes other than CO did not undergo a red shift, indicating that AB1-CO and AB2-CO have high selectivity for CO.

[0103] The present invention has been described in detail above with reference to the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of the present invention.

Claims

1. An Aza-BODIPY type compound, characterized in that, It has the structure shown in Equation I: ； Wherein, R is selected from -(CH2CH2O). n CH3, n≥1; R1 and R2 are selected from -CH2(OCH2CH2) m CH3; m≥1.

2. The Aza-BODIPY compound according to claim 1, characterized in that, 1≤n≤20。 3. The Aza-BODIPY compound according to claim 1, characterized in that, 1≤m≤20。 4. The Aza-BODIPY compound according to claim 1, characterized in that, The Aza-BODIPY class compounds are selected from the following structural formulas: 、 、 。 5. The method for preparing Aza-BODIPY-type compounds according to any one of claims 1 to 4, characterized in that, Includes the following steps: The compound 6, allyl chloroformate, organic base, and organic solvent are mixed and reacted to obtain the product. The structural formula of compound 6 is as follows: 。 6. The preparation method according to claim 5, characterized in that, The reaction temperature is -5~5℃.

7. The preparation method according to claim 5, characterized in that, The organic base is selected from at least one of DIPEA, DEA, TEA, and piperidine.

8. The preparation method according to claim 5, characterized in that, The organic solvent is selected from at least one of dichloromethane, tetrahydrofuran, dimethyl sulfoxide, N'N-dimethylformamide, acetonitrile, or ethanol.

9. A fluorescent probe, characterized in that, Including the Aza-BODIPY type compounds as described in any one of claims 1 to 4.

10. The Aza-BODIPY compound according to any one of claims 1 to 4; or the fluorescent probe according to claim 9 in the detection of carbon monoxide for non-diagnostic or therapeutic purposes.

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