Application of near-infrared photosensitizer BOPYIN in preparation of medicine for treating tumors

Abstract

The invention discloses an application of a near-infrared photosensitizer BOPYIN in preparation of a medicine for treating tumors, wherein the tumors comprise cervical cancer. The photosensitizer is prepared by taking a para-substituted derivative of seven-membered boron-dipyrromethene and p-dimethylamino cinnamyl aldehyde as raw materials through catalysis and condensation of piperidine, acetic acid and phosphorus oxychloride, the synthesis method is simple, separation and purification are convenient, and the yield is relatively high. By introducing dimethylamino cinnamyl aldehyde, the absorption and emission spectrum of the molecule is subjected to red shift, which is beneficial for the structure to be well applied to 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 an emerging tumor treatment technology that uses photosensitizers to produce reactive oxygen species (ROS) under light irradiation of a specific wavelength, thereby killing tumor cells. PDT has the advantages of being non-invasive, highly selective, and non-resistant, and has shown great application potential in tumor treatment. However, traditional photosensitizers have some limitations in their application. For example, under hypoxic conditions in the tumor microenvironment, the oxygen produced by the photosensitizer is consumed too quickly, affecting the therapeutic effect. In addition, the therapeutic effect of PDT is also limited by the accumulation efficiency of the photosensitizer in the tumor tissue and the depth of light penetration.

[0003] Near-infrared fluorescent dyes (NIR) have become ideal photosensitizer materials in photodynamic therapy because of their deeper penetration and less autofluorescence interference in biological tissues. In recent years, researchers have developed a series of NIR fluorescent dyes with excellent performance 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 new ratiometric NIR-II fluorescent organic nanoprobe BTz-IC@IR1061 was developed for tumor-activated photodynamic therapy. The probe activates photodynamic therapy through specific reactions and monitors the treatment process using fluorescence imaging, showing excellent photostability and therapeutic effects.

[0004] Hela cells are a human cervical cancer cell line widely used in tumor research, which is highly proliferative and invasive. In the treatment of Hela tumors, the development of a NIR fluorescent dye that can efficiently accumulate in tumor tissues and 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 used in photodynamic therapy, research on specific treatment and real-time imaging monitoring of Hela tumors is still in the development stage. Therefore, the development of a new type of NIR fluorescent dye for photodynamic therapy of Hela tumors has important scientific significance and clinical application value. Summary of the invention

[0005] The main purpose of the present invention is to provide a preparation method of a near-infrared photosensitizer BOPYIN and its phototherapy application. The technical solution of the present invention is as follows: A preparation method of a near-infrared photosensitizer BOPYIN and its phototherapy application, wherein the chemical structural formula of the compound is: .

[0006] The substituent R is selected from any one of hydrogen, methoxy, bromine or cyano. The chemical structural formula of the compound is: Any one of .

[0007] The synthesis method of the near-infrared photosensitizer BOPYIN comprises the following synthesis path: .

[0008] The method comprises the following steps: (1) Add compound 1, compound 2, and toluene to a reaction bottle at room temperature, dissolve them by ultrasonication, add piperidine, and heat to obtain a reaction solution; (2) The reaction solution in step (1) is subjected to rotary evaporation, and then separated by silica gel column chromatography to obtain product Y. Compound 1 is a seven-membered fluoroborane dipyrrole derivative, and compound 2 is p-dimethylaminocinnamaldehyde; the molar ratio of compound 1 to compound 2 is 1:1-2.

[0009] The order of feeding in the step (1) is compound 1, compound 2, toluene, and piperidine; the feeding ratio of compound 1 to piperidine is 1:0.3~1.

[0010] The feeding sequence and ratio are optimized and designed. Adding the reactants (i.e., compound 1 and compound 2) first can ensure their full contact, adding the solvent later is conducive to controlling the reaction conditions, and adding the catalyst last can effectively regulate the reaction rate. The feeding ratio range of 1:0.3~1 can ensure complete reaction and avoid side reactions. Changing the feeding sequence or ratio may lead to problems such as incomplete reaction, increased by-products or out-of-control reaction.

[0011] The heating temperature of step (1) is 20-100° C. and the heating time is 2-5 hours.

[0012] The beneficial effects of the present invention are as follows: (1) The compound of the present invention is a near-infrared photosensitizer. The phototoxicity experiment verified that the photosensitizer can efficiently generate reactive oxygen species (ROS) under light irradiation and exhibit significant phototoxicity to Hela tumor cells. In addition, the BOPYIN photosensitizer has low toxicity to normal cells under non-light conditions and exhibits good biocompatibility, which is beneficial to improving the safety and selectivity of phototherapy. This shows that BOPYIN has great application potential in the field of photodynamic therapy (PDT).

[0013] (2) The synthetic reaction conditions of the near-infrared photosensitizer BOPYIN described in the present invention are easy to control, the product purification operation is simple, and it has universal applicability. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 It is the hydrogen spectrum of compound Y-1 obtained in Example 1.

[0015] Figure 2 It is the hydrogen spectrum of compound Y-2 obtained in Example 5.

[0016] Figure 3 It is the hydrogen spectrum of compound Y-3 obtained in Example 6.

[0017] Figure 4 It is the hydrogen spectrum of compound Y-4 obtained in Example 7.

[0018] Figure 5 It is the ultraviolet absorption and fluorescence spectrum of compound Y-1 obtained in Example 1.

[0019] Figure 6 It is the ultraviolet absorption and fluorescence spectrum of compound Y-2 obtained in Example 5.

[0020] Figure 7 It is the ultraviolet absorption and fluorescence spectrum of compound Y-3 obtained in Example 6.

[0021] Figure 8 It is the ultraviolet absorption and fluorescence spectrum of compound Y-4 obtained in Example 7.

[0022] Fig. 9 This is a live-dead staining image of compound Y-4 obtained in Example 8. DETAILED DESCRIPTION

[0023] The present invention is further described below with reference to embodiments, but the scope of protection claimed by the present invention is not limited to the scope described in the embodiments.

[0024] Example 1 Compound 1 heptafluoroboranedipyrrole compound (298 mg, 1 mmol) and 4-dimethylaminocinnamaldehyde (175 mg, 1 mmol) were weighed and dissolved in 20 mL of toluene. Piperidine (59 μL, 0.6 mmol) was added and heated in an oil bath at 100 °C with stirring for 5 hours until the reaction was complete. The reaction solution was spin-dried and purified by column chromatography to obtain a purple-black solid Y-1 (218.4 mg) with a yield of 48%.

[0025] .

[0026] Example 2 Compound 1 heptafluoroboranedipyrrole (298 mg, 1 mmol) and 4-dimethylaminocinnamaldehyde (175 mg, 1 mmol) were weighed and dissolved in 20 mL of toluene. Piperidine (99 μL, 1 mmol) was added and heated and stirred in an oil bath at 100° C. for 5 hours until the reaction was complete. The reaction solution was spin-dried and purified by column chromatography to obtain a purple-black solid Y-1 (218.4 mg) with a yield of 42%. When the amount of piperidine was increased by 0.4 mmol relative to Example 1, the yield decreased by 6%.

[0027] .

[0028] Example 3 Compound 1 heptafluoroborane dipyrrole (298 mg, 1 mmol) and 4-dimethylaminocinnamaldehyde (175 mg, 1 mmol) were weighed and dissolved in 20 mL of toluene. Piperidine (29 μL, 0.3 mmol) was added and heated and stirred in an oil bath at 100° C. for 5 hours until the reaction was complete. The reactant was spin-dried and purified by column chromatography to obtain a purple-black solid Y-1 (218.4 mg) with a yield of 36%. When the amount of piperidine was reduced by 0.3 mmol relative to Example 1, the yield was reduced by 12%.

[0029] .

[0030] Example 4 Compound 1 heptafluoroborane dipyrrole (298 mg, 1 mmol) and 4-dimethylaminocinnamaldehyde (262 mg, 1.5 mmol) were weighed and dissolved in 20 mL of toluene, and piperidine (59 μL, 0.6 mmol) was added. The mixture was heated and stirred at 100° C. in an oil bath for 5 hours until the reaction was complete. The reactant was spin-dried and purified by column chromatography to obtain a purple-black solid Y-1 (218.4 mg) with a yield of 46%. When the amount of 4-dimethylaminocinnamaldehyde was increased by 0.5 mmol relative to Example 1, the yield did not change significantly.

[0031] The black solid Y-1 (4.55 mg, 0.01 mmol) was dissolved in 1 mL of dichloromethane to prepare a mother solution. 6 μL of the mother solution was added to 3 mL of toluene, dichloromethane, tetrahydrofuran, acetone, acetonitrile, and dimethyl sulfoxide to measure the absorption wavelength of the compound ( Figure 5 ). Use the maximum absorption wavelength to excite the compound to obtain its emission wavelength ( Figure 5 ). This compound has a long absorption wavelength, with an absorption peak between 400-700nm. Irradiation with laser light in this wavelength band will produce singlet oxygen (ROS), which is beneficial for its application in the field of photodynamic therapy.

[0032] .

[0033] Example 5 Compound 1 heptafluoroboranedipyrrole (328 mg, 1 mmol) and 4-dimethylaminocinnamaldehyde (210 mg, 1.2 mmol) were weighed and dissolved in 20 mL of toluene. Piperidine (99 μL, 1 mmol) was added and heated in an oil bath at 100 °C with stirring for 3 hours until the reaction was complete. The reactants were rotary evaporated and purified by column chromatography to obtain a purple-black solid Y-2 (203.7 mg) with a yield of 42%.

[0034] The black solid Y-2 (4.85 mg, 0.01 mmol) was dissolved in 1 mL of dichloromethane to prepare a mother solution. 6 μL of the mother solution was added to 3 mL of toluene, dichloromethane, tetrahydrofuran, acetone, acetonitrile, and dimethyl sulfoxide to measure the absorption wavelength of the compound ( Figure 6 ). Use the maximum absorption wavelength to excite the compound to obtain its emission wavelength ( Figure 6 ). This compound has a long absorption wavelength, with an absorption peak between 400-700nm. Irradiation with laser light in this wavelength band will produce singlet oxygen (ROS), which is beneficial for its application in the field of photodynamic therapy.

[0035] .

[0036] Example 6 Compound 1 heptafluoroboranedipyrrole (377 mg, 1 mmol) and 4-dimethylaminocinnamaldehyde (210 mg, 1.2 mmol) were weighed and dissolved in 20 mL of toluene. Piperidine (59 μL, 0.6 mmol) was added and heated in an oil bath at 100 °C with stirring for 3 hours until the reaction was complete. The reactant was rotary evaporated and purified by column chromatography to obtain a purple-black solid Y-3 (224.3 mg) with a yield of 42%.

[0037] The black solid Y-3 (5.34 mg, 0.01 mmol) was dissolved in 1 mL of dichloromethane to prepare a mother solution. 6 μL of the mother solution was added to 3 mL of toluene, dichloromethane, tetrahydrofuran, acetone, acetonitrile, and dimethyl sulfoxide to measure the absorption wavelength of the compound ( Figure 7 ). Use the maximum absorption wavelength to excite the compound to obtain its emission wavelength ( Figure 7 ). This compound has a long absorption wavelength, with an absorption peak between 400-700nm. Irradiation with laser light in this wavelength band will produce singlet oxygen (ROS), which is beneficial for its application in the field of photodynamic therapy.

[0038] .

[0039] Example 7 Compound 1 heptafluoroboranedipyrrole (323 mg, 1 mmol) and 4-dimethylaminocinnamaldehyde (210 mg, 1.2 mmol) were weighed and dissolved in 20 mL of toluene. Piperidine (99 μL, 1 mmol) was added and heated in an oil bath at 100°C for 3 hours with stirring until the reaction was complete. The reactant was rotary evaporated and purified by column chromatography to obtain a purple-black solid Y-4 (190 mg) with a yield of 40%.

[0040] The black solid Y-4 (4.80 mg, 0.01 mmol) was dissolved in 1 mL of dichloromethane to prepare a mother solution. 6 μL of the mother solution was added to 3 mL of toluene, dichloromethane, tetrahydrofuran, acetone, acetonitrile, and dimethyl sulfoxide to measure the absorption wavelength of the compound ( Figure 8 ). Use the maximum absorption wavelength to excite the compound to obtain its emission wavelength ( Figure 8 ). This compound has a long absorption wavelength, with an absorption peak between 400-700nm. Irradiation with laser light in this wavelength band will produce singlet oxygen (ROS), which is beneficial for its application in the field of photodynamic therapy.

[0041] .

[0042] Example 8 Cell live-dead staining experiment of compound Y-4 The cell activity detection experiment was carried out using the Hela cell line. Cells in good growth condition were placed at 1.0×10 5 The cells were seeded in six-well plates at a density of 10 cells / mL and incubated at 37°C and 5% CO 2 The cells were incubated overnight in an incubator. Four treatment groups were set up in the experiment: (1) control group (fresh culture medium only); (2) light-only group (635 nm, 1.0 W / cm 2 ); (3) Y-4 treatment group alone (medium containing Y-4); (4) Y-4 combined with light treatment group. After replacing the corresponding medium, the cells were incubated for another 4 hours. Subsequently, the cells in each group were stained with 10 μL Calein-AM (1.0 mM, live cell staining) and 10 μL PI (1.0 mM, dead cell staining), respectively. After staining, the cells in groups 2 and 4 were irradiated with 635 nm laser (1.0 W / cm 2 ). After all groups were incubated for 30 minutes, they were observed and photographed using a fluorescence microscope (Olympus, BX51). The experimental results showed that the control group had the highest cell survival rate; a small amount of cell death occurred in the light-only group; some cell death occurred in the Y-4-only treatment group; and 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.

[0043] The above-mentioned embodiments are only preferred technical solutions of the present invention and should not be regarded as limitations of the present invention. The embodiments and features in the embodiments of the present application can be arbitrarily combined with each other without conflict. The protection scope of the present invention shall 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, 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 structure of the near-infrared photosensitizer BOPYIN is: Wherein, the substituent R is selected from any one of hydrogen, methoxy, bromine or cyano.

2. The use according to claim 1, characterized in that The tumors include cervical cancer.

3. The use according to claim 1, characterized in that The near-infrared photosensitizer BOPYIN is used in preparing drugs for treating tumors under visible light irradiation.

4. The use according to claim 3, characterized in that The wavelength of the visible light is 460-700nm.

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