Use of a near-infrared photosensitizer BOPYIN in the preparation of a drug for treating tumors

Abstract

The present invention discloses the use of a near-infrared photosensitizer BOPYIN in the preparation of a drug for treating tumors, and the tumors include cervical cancer. The photosensitizer is obtained by using a para-substituted derivative of boron dipyrromethene heptamer and p-dimethylaminocinnamaldehyde as raw materials, and through catalysis and condensation with piperidine, acetic acid, and phosphorus oxychloride. The synthesis method is simple, the separation and purification are convenient, and the yield is relatively high. The introduction of p-dimethylaminocinnamaldehyde causes the absorption and emission spectra of the molecule to redshift, which is beneficial to the good application of this structure in photodynamic therapy.

Description

Technical Field

[0001] The present invention relates to a near-infrared photosensitizer, and more specifically, to the preparation and phototherapy application of a near-infrared photosensitizer BOPYIN. Background Art

[0002] Photodynamic Therapy (PDT) is a new emerging tumor treatment technology. By irradiating a photosensitizer with light of a specific wavelength, Reactive Oxygen Species (ROS) are generated, thereby killing tumor cells. PDT has the advantages of non-invasiveness, high selectivity, and no drug resistance, and shows great application potential especially in tumor treatment. However, traditional photosensitizers have some limitations in application. For example, under hypoxic conditions in the tumor microenvironment, the oxygen produced by the photosensitizer is consumed too quickly, affecting the treatment effect. In addition, the treatment effect of PDT is also limited by the accumulation efficiency of the photosensitizer in tumor tissues and the light penetration depth.

[0003] Near-infrared fluorescent dyes (NIR) have become ideal photosensitizer materials in photodynamic therapy due to their deeper penetration in biological tissues and less autofluorescence interference. In recent years, researchers have developed a series of NIR fluorescent dyes with excellent properties through molecular structure design and nanotechnology. These dyes can not only achieve efficient photodynamic therapy but also monitor the treatment process in real time through fluorescence imaging. For example, a novel ratiometric NIR-II fluorescent organic nanoprobe BTz-IC@IR1061 was developed for tumor-activated photodynamic therapy. This probe activates photodynamic therapy through a specific reaction and monitors the treatment process using fluorescence imaging, showing excellent photostability and treatment effect.

[0004] Hela cells are a human cervical cancer cell line widely used in tumor research, with high proliferative and invasive properties. In the treatment of Hela tumors, developing a NIR fluorescent dye that can efficiently accumulate in tumor tissues, has good photostability and biocompatibility is of great significance for improving the effect of photodynamic therapy. At present, although a variety of NIR fluorescent dyes have been applied to photodynamic therapy, the research on specific treatment and real-time imaging monitoring for Hela tumors is still in the development stage. Therefore, developing a new type of NIR fluorescent dye for the photodynamic therapy of Hela tumors has important scientific significance and clinical application value. Summary of the Invention

[0005] The main object of the present invention is to provide the preparation and phototherapy application of a near-infrared photosensitizer BOPYIN. The technical solution of the present invention is as follows:

[0006] Preparation of a near-infrared photosensitizer BOPYIN and its phototherapy applications. The chemical structural formula of the compound is:

[0007] 。

[0008] The substituent R is selected from any one of hydrogen, methoxy, bromine or cyano. The chemical structural formula of the compound is:

[0009] any one of

[0010] The synthesis method of the near-infrared photosensitizer BOPYIN. The method includes the following synthetic route:

[0011] 。

[0012] The method includes the following steps:

[0013] (1) Add compound 1, compound 2, and toluene to a reaction flask at room temperature. After ultrasonic dissolution, add piperidine, and heat to obtain a reaction solution;

[0014] (2) Rotavaporize the reaction solution in step (1), and then separate it by silica gel column chromatography to obtain product Y. Compound 1 is a heptafluoroboron dipyrrole derivative, and compound 2 is p-dimethylaminocinnamaldehyde; the molar ratio of compound 1 to compound 2 in the feed is 1:1~2.

[0015] The feeding order in step (1) is compound 1, compound 2, toluene, piperidine; the feeding ratio of compound 1 to piperidine is 1:0.3~1.

[0016] The feeding order and ratio are optimized. Adding the reactants (i.e., compound 1 and compound 2) first can ensure their full contact. Adding the solvent later is beneficial for controlling the reaction conditions. Adding the catalyst finally can effectively regulate the reaction rate. And the feeding ratio range of 1:0.3~1 can not only ensure the complete reaction but also avoid side reactions. Changing the feeding order or ratio may lead to problems such as incomplete reaction, increased by-products or out-of-control reaction.

[0017] The heating temperature in step (1) is 20~100 °C, and the heating time is 2~5 hours.

[0018] The beneficial effects of the present invention are as follows:

[0019] (1) The compound of the present invention is a near-infrared photosensitizer. Through phototoxicity experiments, it is verified that this photosensitizer can efficiently generate reactive oxygen species (ROS) under light irradiation and exhibits significant phototoxicity to Hela tumor cells. Moreover, this BOPYIN photosensitizer has low toxicity to normal cells under dark conditions, showing good biocompatibility, which is beneficial to improving the safety and selectivity of phototherapy. This indicates that BOPYIN has great application potential in the field of photodynamic therapy (PDT).

[0020] (2) The synthesis reaction conditions of the near-infrared photosensitizer BOPYIN of the present invention are easy to control, and the product purification operation is simple, with universal applicability. Brief Description of the Drawings

[0021] Figure 1 It is the hydrogen spectrum of compound Y-1 obtained in Example 1.

[0022] Figure 2 It is the hydrogen spectrum of compound Y-2 obtained in Example 5.

[0023] Figure 3 It is the hydrogen spectrum of compound Y-3 obtained in Example 6.

[0024] Figure 4 It is the hydrogen spectrum of compound Y-4 obtained in Example 7.

[0025] Figure 5 It is the ultraviolet absorption and fluorescence spectrum of compound Y-1 obtained in Example 1.

[0026] Figure 6 It is the ultraviolet absorption and fluorescence spectrum of compound Y-2 obtained in Example 5.

[0027] Figure 7 It is the ultraviolet absorption and fluorescence spectrum of compound Y-3 obtained in Example 6.

[0028] Figure 8 It is the ultraviolet absorption and fluorescence spectrum of compound Y-4 obtained in Example 7.

[0029] Figure 9 It is the live / dead cell staining image of compound Y-4 obtained in Example 8. Detailed Embodiments

[0030] The following examples are used to further illustrate the present invention, but the scope claimed by the present invention is not limited to the scope described in the examples.

[0031] Example 1

[0032] Weigh the heptacyclic boron difluoride dipyrromethene compound of Compound 1 (298 mg, 1 mmol), 4-dimethylaminocinnamaldehyde (175 mg, 1 mmol), dissolve them in 20 mL of toluene, then add piperidine (59 μL, 0.6 mmol), heat and stir at 100 °C in an oil bath for 5 hours until the reaction is complete. Rotate the reaction solution to dryness, and after purification by column chromatography, obtain a purple-black solid Y-1 (218.4 mg) with a yield of 48%.

[0033] 。

[0034] Example 2

[0035] Weigh the heptacyclic boron difluoride dipyrromethene compound of Compound 1 (298 mg, 1 mmol), 4-dimethylaminocinnamaldehyde (175 mg, 1 mmol), dissolve them in 20 mL of toluene, then add piperidine (99 μL, 1 mmol), heat and stir at 100 °C in an oil bath for 5 hours until the reaction is complete. Rotate the reaction solution to dryness, and after purification by column chromatography, obtain a purple-black solid Y-1 (218.4 mg) with a yield of 42%. When the amount of piperidine is increased by 0.4 mmol relative to Example 1, the yield is reduced by 6%.

[0036] 。

[0037] Example 3

[0038] Weigh the heptacyclic boron difluoride dipyrromethene compound of Compound 1 (298 mg, 1 mmol), 4-dimethylaminocinnamaldehyde (175 mg, 1 mmol), dissolve them in 20 mL of toluene, then add piperidine (29 μL, 0.3 mmol), heat and stir at 100 °C in an oil bath for 5 hours until the reaction is complete. Rotate the reaction mixture to dryness, and after purification by column chromatography, obtain a purple-black solid Y-1 (218.4 mg) with a yield of 36%. When the amount of piperidine is decreased by 0.3 mmol relative to Example 1, the yield is reduced by 12%.

[0039] 。

[0040] Example 4

[0041] Weigh the heptacyclic boron difluoride dipyrromethene compound of Compound 1 (298 mg, 1 mmol), 4-dimethylaminocinnamaldehyde (262 mg, 1.5 mmol), dissolve them in 20 mL of toluene, then add piperidine (59 μL, 0.6 mmol), heat and stir at 100 °C in an oil bath for 5 hours until the reaction is complete. Rotate the reaction mixture to dryness, and after purification by column chromatography, obtain a purple-black solid Y-1 (218.4 mg) with a yield of 46%. When the amount of 4-dimethylaminocinnamaldehyde is increased by 0.5 mmol relative to Example 1, the yield shows no obvious change.

[0042] The black solid Y-1 (4.55 mg, 0.01 mmol) was dissolved in 1 mL of dichloromethane to prepare a mother liquor. 6 μL of the mother liquor was added to 3 mL of toluene, dichloromethane, tetrahydrofuran, acetone, acetonitrile, and dimethyl sulfoxide respectively, and the absorption wavelength of the compound was measured ( Figure 5 ). The obtained maximum absorption wavelength was used to excite the compound to obtain its emission wavelength ( Figure 5 ). The compound has a relatively long absorption wavelength, with an absorption peak between 400 - 700 nm. Irradiating with a laser lamp in this wavelength band will cause it to generate singlet oxygen (ROS), which is beneficial for its application in the field of photodynamic therapy.

[0043] .

[0044] Example 5

[0045] Weigh the heptacyclic fluoroboron dipyrromethene compound of Compound 1 (328 mg, 1 mmol) and 4-dimethylaminocinnamaldehyde (210 mg, 1.2 mmol), dissolve them in 20 mL of toluene, then add piperidine (99 μL, 1 mmol), heat and stir at 100 °C in an oil bath for 3 hours until the reaction is complete. Rotate evaporate the reactants and purify by column chromatography to obtain a purple-black solid Y-2 (203.7 mg) with a yield of 42%.

[0046] The black solid Y-2 (4.85 mg, 0.01 mmol) was dissolved in 1 mL of dichloromethane to prepare a mother liquor. 6 μL of the mother liquor was added to 3 mL of toluene, dichloromethane, tetrahydrofuran, acetone, acetonitrile, and dimethyl sulfoxide respectively, and the absorption wavelength of the compound was measured ( Figure 6 ). The obtained maximum absorption wavelength was used to excite the compound to obtain its emission wavelength ( Figure 6 ). The compound has a relatively long absorption wavelength, with an absorption peak between 400 - 700 nm. Irradiating with a laser lamp in this wavelength band will cause it to generate singlet oxygen (ROS), which is beneficial for its application in the field of photodynamic therapy.

[0047] .

[0048] Example 6

[0049] Weigh the heptacyclic fluoroboron dipyrromethene compound of Compound 1 (377 mg, 1 mmol) and 4-dimethylaminocinnamaldehyde (210 mg, 1.2 mmol), dissolve them in 20 mL of toluene, then add piperidine (59 μL, 0.6 mmol), heat and stir at 100 °C in an oil bath for 3 hours until the reaction is complete. Rotate evaporate the reactants and purify by column chromatography to obtain a purple-black solid Y-3 (224.3 mg) with a yield of 42%.

[0050] The black solid Y-3 (5.34 mg, 0.01 mmol) was dissolved in 1 mL of dichloromethane to prepare a stock solution. 6 μL of the stock solution was added to 3 mL of toluene, dichloromethane, tetrahydrofuran, acetone, acetonitrile, and dimethyl sulfoxide respectively, and the absorption wavelength of the compound was measured ( Figure 7 ). The maximum absorption wavelength obtained was used to excite the compound to obtain its emission wavelength ( Figure 7 ). The compound has a relatively long absorption wavelength, with an absorption peak between 400 - 700 nm. Irradiating with a laser lamp in this wavelength band will cause it to generate singlet oxygen (ROS), which is beneficial for its application in the field of photodynamic therapy.

[0051] .

[0052] Example 7

[0053] Weighed 1 mmol (323 mg) of the heptafluoroboron dipyrromethene compound of Compound 1 and 1.2 mmol (210 mg) of 4-dimethylaminocinnamaldehyde, dissolved them in 20 mL of toluene, then added piperidine (99 μL, 1 mmol), and heated and stirred at 100 °C in an oil bath for 3 hours until the reaction was complete. The reaction product was rotary evaporated and purified by column chromatography to obtain a purple-black solid Y-4 (190 mg), with a yield of 40%.

[0054] The black solid Y-4 (4.80 mg, 0.01 mmol) was dissolved in 1 mL of dichloromethane to prepare a stock solution. 6 μL of the stock solution was added to 3 mL of toluene, dichloromethane, tetrahydrofuran, acetone, acetonitrile, and dimethyl sulfoxide respectively, and the absorption wavelength of the compound was measured ( Figure 8 ). The maximum absorption wavelength obtained was used to excite the compound to obtain its emission wavelength ( Figure 8 ). The compound has a relatively long absorption wavelength, with an absorption peak between 400 - 700 nm. Irradiating with a laser lamp in this wavelength band will cause it to generate singlet oxygen (ROS), which is beneficial for its application in the field of photodynamic therapy.

[0055] .

[0056] Example 8 Cell Viability and Cytotoxicity Staining Experiment of Compound Y-4

[0057] The cell viability detection experiment was carried out using the Hela cell line. Cells in good growth state were seeded in a six-well plate at a density of 1.0×10 5 cells per well and incubated overnight in an incubator at 37 °C and 5% CO2. Four groups of treatments were set up in the experiment: (1) Control group (only fresh medium); (2) Simple light irradiation group (635 nm, 1.0 W / cm 2); (3) simple Y-4 treatment group (culture medium containing Y-4); (4) Y-4 combined with light treatment group. After changing the corresponding culture medium, continue to incubate for 4 hours. Subsequently, the cells in each group were stained with 10 μL of Calein-AM (1.0 mM, live cell staining) and 10 μL of PI (1.0 mM, dead cell staining), and the cells in the second and fourth groups were irradiated with 635 nm laser (1.0 W / cm 2 ). After all groups were incubated for another 30 minutes, they were observed and photographed using a fluorescence microscope (Olympus, BX51). The experimental results showed that the cell survival rate was the highest in the control group; a small number of cells died in the simple light treatment group; partial cell death occurred in the simple Y-4 treatment group; while in the Y-4 combined with light treatment group, almost all cancer cells were killed, indicating that Y-4 has a significant photodynamic therapy effect under light conditions.

[0058] The above embodiments are only the preferred technical solutions of the present invention and should not be regarded as limitations to the present invention. The embodiments in this application and the features in the embodiments can be arbitrarily combined with each other without conflict. The protection scope of the present invention should be the technical solutions recorded in the claims, including the equivalent replacement solutions of the technical features in the technical solutions recorded in the claims. That is, the equivalent replacement improvements within this scope are also within the protection scope of the present invention.

Claims

1. Use of a near-infrared photosensitizer BOPYIN in the preparation of a drug for treating tumors, characterized in that, The chemical structural formula of the near-infrared photosensitizer BOPYIN is as follows: Among them, the substituent R is selected from any one of hydrogen, methoxy, bromine or cyano, and the tumor is cervical cancer.

2. The use according to claim 1, wherein The near-infrared photosensitizer BOPYIN is used in the preparation of drugs for treating tumors under visible light irradiation.

3. The use according to claim 2, wherein The wavelength of the visible light is 460 - 700 nm.

Images