A triphenylethylene-xanthene derivative, preparation method and viscosity detection fluorescent probe

Abstract

This invention relates to the field of optical sensing and imaging detection technology, specifically to a triphenylene-oxanthracene derivative, its preparation method, and a fluorescent probe for viscosity detection. The triphenylene-oxanthracene derivative has the general formula shown in Formula I. Compared to Rhodamine B, the synthesized triphenylene-oxanthracene derivative exhibits a larger Stokes shift. Furthermore, the fluorescence intensity of the triphenylene-oxanthracene derivative increases with increasing cell viscosity. In addition, the triphenylene-oxanthracene derivative can be further used to detect mitochondrial viscosity, exhibiting good mitochondrial targeting. In summary, the probe improves the Stokes shift, has a wide optical response pH range, and possesses excellent mitochondrial targeting.

Description

Technical Field

[0001] This invention relates to the field of optical sensing imaging and detection technology, specifically to a triphenylene-oxanthracene derivative, its preparation method, and a viscosity detection fluorescent probe. Background Technology

[0002] Changes in the microenvironment of organelles in biological systems are closely related to cellular function. Viscosity, as an important parameter of the cellular microenvironment, can participate in the regulation of biological processes and reflect the state and function of organelles. Abnormal viscosity values ​​can lead to various diseases, such as atherosclerosis and Alzheimer's disease, while abnormal mitochondrial viscosity may lead to Parkinson's disease. Therefore, monitoring changes in mitochondrial viscosity is of great significance, and developing viscosity detection probes can help elucidate the mechanisms by which abnormal mitochondrial viscosity triggers related diseases and further understand cellular function.

[0003] Fluorescent probes, due to their high selectivity and excellent biocompatibility, can be used for viscosity determination at the subcellular level. Fluorescent molecular rotors are commonly used to detect the viscosity of solvents and biofluids, offering higher accuracy and reliability. However, some current fluorescent probes for viscosity detection suffer from drawbacks such as low photostability, small Stokes shift, and poor organelle targeting, necessitating the design of novel probes for mitochondrial-targeted viscosity monitoring. Summary of the Invention

[0004] The purpose of this invention is to address the problems of poor mitochondrial targeting and small Stokes shift in existing viscosity detection probes, which cannot adequately meet the requirements for photostability, Stokes shift, and organelle targeting in biomolecular detection. This invention provides a triphenylene-oxanthracene derivative, its preparation method, and a fluorescent probe for viscosity detection.

[0005] The first aspect of this invention provides a triphenylene-oxanthracene derivative having the general formula shown in Formula I:

[0006]

[0007] R1, R2, and R3 are each independently selected from C1-C4 alkyl groups; R4, R5, and R6 are each independently selected from hydrogen, deuterium, and C1-C4 alkyl groups.

[0008] Preferably, R1, R2, and R3 are each independently selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl. R4, R5, and R6 are each independently selected from hydrogen, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl.

[0009] Preferably, the triphenylene-oxanthracene derivative is selected from any one of the following:

[0010]

[0011] A second aspect of this invention provides a method for preparing the above-mentioned triphenylene-oxanthracene derivative, wherein the compound of formula I-1 is reacted with the compound of formula I-2 to obtain the triphenylene-oxanthracene derivative, and the reaction equation is as follows:

[0012]

[0013] Wherein, R1, R2, R3, R4, R5, and R6 are as defined in the first aspect of this invention.

[0014] Preferably, the procedure specifically includes the following steps:

[0015] 1) The compound of formula I-1 and the compound of formula I-2 are reacted in the first solvent to obtain the intermediate;

[0016] 2) The intermediate provided in step 1) is refluxed in the presence of a second solvent and a catalyst, and then post-treated to obtain the triphenylene-oxanthracene derivative.

[0017] Preferably, it includes at least one of the following technical features:

[0018] In step 1), the molar ratio of compound I-1 to compound I-2 is 1:(1-1.5).

[0019] A2) In step 1), the reaction temperature is 90℃-130℃;

[0020] In step 1) of A3), the reaction time is 8h-16h.

[0021] Preferably, it includes at least one of the following technical features:

[0022] B1) The first solvent is selected from one or more of concentrated sulfuric acid and methanesulfonic acid;

[0023] B2) The second solvent is selected from one or more of methanol and ethanol;

[0024] B3) The catalyst is selected from concentrated sulfuric acid;

[0025] B4) The mass-to-volume ratio of the I-1 compound to the first solvent is (1.5–2) g : (2–8) mL;

[0026] B5) The mass-to-volume ratio of the I-1 compound to the second solvent is (1.5–2) g : (20–40) mL;

[0027] B6) The mass-to-volume ratio of the I-1 compound to the catalyst is (1.5–2) g : (1–3) mL;

[0028] B7) The post-processing steps include cooling the solution after the reaction, distillation, neutralization to neutral, extraction and combining of organic solvents, drying, distillation, and chromatography.

[0029] Preferably, in feature B7), the extraction solvent is dichloromethane.

[0030] Preferably, in feature B7), the mobile phase of the chromatography is dichloromethane and methanol; the volume ratio of dichloromethane to ethyl acetate is 20 / 1 to 10 / 1.

[0031] The third aspect of this invention provides the use of the above-mentioned triphenylene-oxanthracene derivative in the preparation of a viscosity detection fluorescent probe.

[0032] A fourth aspect of the present invention provides a viscosity detection fluorescent probe, comprising the triphenylene-oxanthracene derivative described in the first aspect of the present invention.

[0033] Preferably, the detection solvent system of the viscosity detection fluorescent probe is selected from either an alcohol-water solution or an acetonitrile-water solution.

[0034] The present invention has the following beneficial effects:

[0035] Compared to Rhodamine B, synthesized triphenylxanthracene derivatives (e.g., the dye TRMe) exhibit a larger Stokes shift. Furthermore, the fluorescence intensity of these derivatives increases with increasing cellular viscosity. Additionally, the triphenylxanthracene derivatives (e.g., the dye TRMe) can be further used to detect mitochondrial viscosity, demonstrating good mitochondrial targeting. In summary, the probe exhibits improved Stokes shift, a wide optical response pH range, and excellent mitochondrial targeting. Attached Figure Description

[0036] Figure 1 This is the MS spectrum of the compound from Example 1.

[0037] Figure 2 It is the compound of Example 1. 1 H NMR spectrum.

[0038] Figure 3 It is the compound of Example 1. 13 C NMR spectrum.

[0039] Figure 4 This is the fluorescence spectrum of the probe in Example 3 at different viscosities of glycerol / H2O.

[0040] Figure 5 This is the linearity calculation of the probe at 632 nm in different viscosities of glycerol / H2O in Example 3.

[0041] Figure 6 These are the fluorescence emission spectra of the probe in different solvent systems in Example 4.

[0042] Figure 7 These are the UV absorption spectra of the probe in different solvent systems in Example 5. Detailed Implementation

[0043] The following specific examples illustrate the implementation of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention.

[0044] It should be noted that the process equipment or apparatus not specifically mentioned in the following embodiments are all conventional equipment or apparatus in the art.

[0045] Triphenylene-oxanthracene derivatives

[0046] This invention provides a triphenylene-oxanthracene derivative having the general formula shown in Formula I:

[0047]

[0048] R1, R2, and R3 are each independently selected from C1-C4 alkyl groups; R4, R5, and R6 are each independently selected from hydrogen, deuterium, and C1-C4 alkyl groups.

[0049] Optionally, R1, R2, and R3 are each independently selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl. R4, R5, and R6 are each independently selected from hydrogen, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl.

[0050] In a preferred embodiment, R1, R2, and R3 are each independently methyl or ethyl. R4, R5, and R6 are each independently selected from hydrogen, methyl, and ethyl.

[0051] In a preferred embodiment, the triphenylene-oxanthracene derivative is selected from any one of the following:

[0052]

[0053]

[0054] Preparation method of triphenylene-oxanthracene derivatives

[0055] A second aspect of this invention provides a method for preparing the above-mentioned triphenylene-oxanthracene derivative, wherein the compound of formula I-1 is reacted with the compound of formula I-2 to obtain the triphenylene-oxanthracene derivative, and the reaction equation is as follows:

[0056]

[0057] R1, R2, and R3 are each independently selected from C1-C4 alkyl groups, and R4, R5, and R6 are each independently selected from hydrogen, deuterium, and C1-C4 alkyl groups.

[0058] Optionally, R1, R2, and R3 are each independently selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl, and R4, R5, and R6 are each independently selected from hydrogen, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl.

[0059] In a preferred embodiment, R1, R2, and R3 are each independently methyl or ethyl, and R4, R5, and R6 are each independently selected from hydrogen, methyl, and ethyl.

[0060] In a preferred embodiment, the steps include:

[0061] 1) The compound of formula I-1 and the compound of formula I-2 are reacted in the first solvent to obtain the intermediate;

[0062] 2) The intermediate provided in step 1) is refluxed in the presence of a second solvent and a catalyst, and then post-treated to obtain the triphenylene-oxanthracene derivative.

[0063] In the preparation method of the triphenylene-oxanthracene derivative provided by the present invention, in step 1), the molar ratio of compound I-1 to compound I-2 is 1:(1-1.5). Optionally, the molar ratio of compound I-1 to compound I-2 is 1:(1-1.2) or 1:(1.2-1.5), etc.

[0064] In the preparation method of the triphenylene-oxanthracene derivative provided by the present invention, in step 1), the reaction temperature is 90℃-130℃. Optionally, the reaction temperature can be, for example, 90℃-120℃, 120℃-130℃, 90℃-100℃, 100℃-110℃, 110℃-120℃, 80℃-100℃, or 100℃-130℃, etc.

[0065] In the preparation method of the triphenylene-oxanthracene derivative provided by the present invention, in step 1), the reaction time is 8h-16h, and optionally, the reaction time can be, for example, 8h-12h or 12h-16h.

[0066] In the preparation method of the triphenylene-oxanthracene derivative provided by the present invention, in step 1), the first solvent is selected from one or more of concentrated sulfuric acid and methanesulfonic acid.

[0067] In the preparation method of the triphenylene-oxanthracene derivative provided by the present invention, in step 2), the second solvent is selected from one or more of methanol and ethanol.

[0068] In the preparation method of the triphenylene-oxanthracene derivative provided by the present invention, in step 2), the catalyst is selected from concentrated sulfuric acid.

[0069] In the preparation method of the triphenylene-oxanthracene derivative provided by the present invention, the mass-to-volume ratio of the I-1 compound to the first solvent is (1.5-2) g:(2-8) mL. Optionally, the mass-to-volume ratio of the I-1 compound to the first solvent is (1.5-1.8) g:(2-8) mL, (1.8-2) g:(2-8) mL, (1.5-2) g:(2-5) mL, or (1.5-2) g:(5-8) mL, etc.

[0070] In the preparation method of the triphenylene-oxanthracene derivative provided by the present invention, the mass-to-volume ratio of the I-1 compound to the second solvent is (1.5-2) g:(20-40) mL. Optionally, the mass-to-volume ratio of the I-1 compound to the second solvent is (1.5-1.8) g:(20-40) mL, (1.8-2) g:(20-40) mL, (1.5-2) g:(20-30) mL, or (1.5-2) g:(30-40) mL, etc.

[0071] In the method for preparing the triphenylene-oxanthracene derivative provided by this invention, the mass-to-volume ratio of the I-1 compound to the catalyst is (1.5–2) g : (1–3) mL. Optionally, the mass-to-volume ratio of the I-1 compound to the catalyst can be, for example, (1.5–2) g : (1–2) mL, (1.5–2) g : (2–3) mL, 1.5–1.8) g : (1–3) mL, or (1.8–2) g : (1–3) mL, etc.

[0072] In the preparation method of the triphenylene-oxanthracene derivative provided by the present invention, the post-processing steps include cooling the reaction solution, distillation, neutralization to neutrality, extraction and combining organic solvents, drying, distillation, and chromatography.

[0073] In some embodiments, the extraction solvent is dichloromethane.

[0074] In some embodiments, the mobile phase of the chromatography is dichloromethane and methanol; the volume ratio of dichloromethane to ethyl acetate is 20 / 1 to 10 / 1. Optionally, the volume ratio of dichloromethane to ethyl acetate is 20 / 1 to 15 / 1 or 15 / 1 to 10 / 1, etc.

[0075] In a specific embodiment,

[0076]

[0077] Compounds 2 and 3 were mixed thoroughly, and concentrated sulfuric acid of a certain concentration was added. The mixture was heated to react. After the reaction, the mixture was slowly poured into ice water. The purple-red solid was collected, dried, dissolved in methanol, and then refluxed with concentrated sulfuric acid as a catalyst. After the solution cooled, it was distilled under reduced pressure. The residue was poured into the ice-water mixture and neutralized with a saturated sodium bicarbonate solution. The mixture was extracted with dichloromethane, and the organic solvents were combined and dried over anhydrous sodium sulfate. After most of the solvent was distilled off under reduced pressure, the product was purified by silica gel column chromatography to obtain the target product.

[0078] Viscosity detection fluorescent probe

[0079] The viscosity detection fluorescent probe of the triphenylene-oxanthracene derivative in this invention includes the above-mentioned triphenylene-oxanthracene derivative, and the detection solvent system of the viscosity detection fluorescent probe is selected from either alcohol-water solution or acetonitrile-water solution.

[0080] The present invention will be further described below with reference to specific embodiments. It should be understood that the following embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or according to the product instructions. All reagents and raw materials used in this invention are commercially available.

[0081] Compound 2 Brand: Aladdin, Name, Catalog Number: M401539-250mg, 1-(4-methoxyphenyl)-1,2,2-triphenylene, CAS: 70592-05-1;

[0082] Compound 3 Brand: Aladdin Product No.: D589552-5g, Cas No.: 5809-23-4.

[0083] Example 1

[0084] The specific method is as follows:

[0085]

[0086] Compound 2 (1.8 g, 5 mmol) and compound 3 (1.6 g, 5 mmol) were mixed thoroughly, and 5 mL of 98% concentrated sulfuric acid was added. The mixture was reacted at 90 °C for 12 hours, and then slowly poured into 150 g of ice water. The purple-red solid was collected, dried, and dissolved in 30 mL of methanol. 2 mL of 98% concentrated sulfuric acid was added as a catalyst, and the mixture was refluxed for 24 hours. After cooling, most of the solvent was evaporated under reduced pressure, and the residue was poured into 100 g of the ice-water mixture. The mixture was neutralized to neutral with saturated sodium bicarbonate solution. Extraction was performed with dichloromethane (50 mL, three times). The organic solvents were combined and dried over anhydrous sodium sulfate. After evaporation of most of the solvent under reduced pressure, the residue was separated by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to obtain 2.4 g of the target product, yield: 75%. MS: 640.2848.

[0087] 1 H NMR(CDCl3,400MHz)8.28(d,J=7.9Hz,1H),7.82(td,J=7.6,1.1Hz,1H),7.73(td,J=7.8,1.1Hz,1H),7.45(dd,J=7.5,2.3Hz,1H),7.3 5-7.29(m,3H),7.17-7.00(m,16H),6.87(d,J=8.5Hz,1H),6.79(d,J=2.3Hz,1H),3.99-3.70(m,4H),3.66(S,3H),1.40-1.35(m,6H);

[0088] 13 C NMR(CDCl3,100MHz)165.41,159.78,159.27,158.51,153.79,153.50,145 .88,142.57,142.47,142.39,138.75,133.55,133.20,132.84,131.48,13 1.30,130.90,130.49,129.80,129.77,128.40,127.93,127.58,127.52,1 19.98,119.90,119.40,118.52,96.45,54.30,52.65,47.46,13.67,12.40;

[0089] MS results are shown below. Figure 1 , 1 H NMR see Figure 2 , 13 C NMR (see) Figure 3 .

[0090] The compound prepared in Example 1 was used as probe 1 for determination in Examples 2-5.

[0091] Example 2: Determination of the optical properties of probe 1 in solutions with different glycerol ratios:

[0092] The specific method is as follows:

[0093] Optical properties determination of triphenylene-oxanthracene derivatives in solutions with different glycerol / H2O ratios: Triphenylene-oxanthracene derivatives were dissolved in glycerol / H2O solvents with glycerol contents of 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 99%, and prepared into solutions with a concentration of 1×10⁻⁶ ppm. -5 3 mL of the test solution with a concentration of mol / L was used to obtain the fluorescence emission spectrum. The excitation wavelength was 520 nm, and the slit width was 10 nm.

[0094] Example 3: Determination of the optical properties of probe 1 in solutions with different glycerol ratios:

[0095] The specific method is as follows:

[0096] Optical properties determination of triphenylene-oxanthracene derivatives in solutions with different glycerol / H2O ratios: Triphenylene-oxanthracene derivatives were dissolved in glycerol / H2O solvents with glycerol contents of 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 99%, and prepared into solutions with a concentration of 1×10⁻⁶ ppm. -5 3 mL of the test solution with a concentration of mol / L was used to obtain the fluorescence emission spectrum, as shown below. Figure 4 As shown. Excitation wavelength 520nm, slit width 10nm, 10nm. Viscosity changes from 1.0cp to 1238.0cp, corresponding to different glycerol contents, as plotted on the x-axis. The fluorescence intensity at 632nm is plotted on the y-axis to calculate the linearity R of the probe's fluorescence intensity at different viscosities. 2 It is 0.999, such as Figure 5 .

[0097] Example 4: Fluorescence emission spectroscopy determination of probe 1 in different solvent systems:

[0098] The specific method is as follows:

[0099] Optical properties determination of triphenylene-oxanthracene derivatives in different solvent systems: Triphenylene-oxanthracene derivatives were dissolved in different solvents, including acetone, ethanol (EtOH), acetonitrile (CH3CN), tetrahydrofuran (THF), water (H2O), dimethylformamide (DMF), dichloromethane (CH2Cl2), and methanol (MeOH), to prepare solutions with a concentration of 1×10⁻⁶. -5 3 mL of the test solution with a concentration of mol / L was used to obtain the fluorescence emission spectrum, as shown below. Figure 6As shown. Excitation wavelength 500nm, slit width 10nm, 10nm. Test results show that fluorescence is strongest in THF and weakest in H2O.

[0100] Example 5: Ultraviolet absorption spectroscopy determination of probe 1 in different solvent systems:

[0101] The specific method is as follows:

[0102] UV absorption spectra of triphenylene-oxanthracene derivatives in different solvent systems: Triphenylene-oxanthracene derivatives were dissolved in different solvents, including acetone, ethanol (EtOH), acetonitrile (CH3CN), tetrahydrofuran (THF), water (H2O), dimethylformamide (DMF), dichloromethane (CH2Cl2), and methanol (MeOH), to prepare solutions of 1×10⁻⁶. -5 3 mL of the test solution with a concentration of mol / L was used to obtain the ultraviolet absorption spectrum, as shown below. Figure 7 As shown in the figure. The test results show that the absorption peaks at 520nm and 550nm are the strongest.

[0103] The above description, in conjunction with specific embodiments, provides a further detailed explanation of the present invention. It should not be construed that the specific implementation of the present invention is limited to these descriptions. For those skilled in the art, several simple deductions or substitutions can be made without departing from the concept of the present invention, and all such deductions or substitutions should be considered within the scope of protection of the present invention. Using the triphenylene-oxanthracene derivative of the present invention as a fluorescent dye is one application; it should not be construed that the triphenylene-oxanthracene derivative of the present invention is only used for fluorescent dyes. For those skilled in the art, considering the same mechanism of action of the triphenylene-oxanthracene derivative of the present invention as a fluorescent dye, several simple inferences can be made to derive other applications of the triphenylene-oxanthracene derivative of the present invention, and all such applications should be considered within the scope of protection of the present invention.

Claims

1. A triphenylene-oxanthracene derivative, characterized in that, It has the general formula shown in Equation I: R1, R2, and R3 are each independently selected from C1-C4 alkyl groups; R4, R5, and R6 are each independently selected from hydrogen, deuterium, and C1-C4 alkyl groups.

2. The triphenylene-oxanthracene derivative according to claim 1, characterized in that, R1, R2, and R3 are each independently selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl. And / or, R4, R5, and R6 are each independently selected from hydrogen, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl.

3. The triphenylene-oxanthracene derivative according to claim 1 or 2, characterized in that, The triphenylene-oxanthracene derivative is selected from any one of the following:

4. The method for preparing the triphenylene-oxanthracene derivative according to any one of claims 1-3, characterized in that, Specifically, the following steps are included: 1) The compound of formula I-1 and the compound of formula I-2 are reacted in the first solvent to obtain the intermediate; 2) The intermediate provided in step 1) is refluxed in the presence of a second solvent and a catalyst, and then post-treated to obtain the triphenylene-oxanthracene derivative. The reaction equation is as follows: R1, R2, R3, R4, R5, and R6 are as defined in claims 1 to 2.

5. The method for preparing the triphenylene-oxanthracene derivative according to claim 4, characterized in that, Includes at least one of the following technical features: In step 1), the molar ratio of compound I-1 to compound I-2 is 1:(1-1.5). A2) In step 1), the reaction temperature is 90℃-130℃; In step 1) of A3), the reaction time is 8h-16h.

6. The method for preparing the triphenylene-oxanthracene derivative according to claim 4, characterized in that, Includes at least one of the following technical features: B1) The first solvent is selected from one or more of concentrated sulfuric acid and methanesulfonic acid; B2) The second solvent is selected from one or more of methanol and ethanol; B3) The catalyst is selected from concentrated sulfuric acid; B4) The mass-to-volume ratio of the I-1 compound to the first solvent is (1.5–2) g : (2–8) mL; B5) The mass-to-volume ratio of the I-1 compound to the second solvent is (1.5–2) g : (20–40) mL; B6) The mass-to-volume ratio of the I-1 compound to the catalyst is (1.5–2) g : (1–3) mL; B7) The post-processing steps include cooling the solution after the reaction, distillation, neutralization to neutral, extraction and combining of organic solvents, drying, distillation, and chromatography.

7. The method for preparing the triphenylene-oxanthracene derivative according to claim 6, characterized in that, Includes at least one of the following technical features: In characteristic B7), the extraction solvent is dichloromethane; In feature B7), the mobile phase of the chromatography is dichloromethane and methanol; the volume ratio of dichloromethane to ethyl acetate is 20 / 1 to 10 / 1.

8. The triphenylene-oxanthracene derivative according to any one of claims 1-3 is used to prepare a viscosity detection fluorescent probe.

9. A fluorescent probe for viscosity detection, characterized in that, Includes the triphenylene-oxanthracene derivatives as described in any one of claims 1-3.

10. The viscosity detection fluorescent probe according to claim 9, characterized in that, The detection solvent system of the viscosity detection fluorescent probe is selected from either alcohol-water solution or acetonitrile-water solution.

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