A photosensitizer, a preparation method and application thereof
By preparing photosensitizer compound C with a longer near-infrared fluorescence emission wavelength and a high singlet oxygen yield, the problem of low efficiency of existing photosensitizers has been solved, enabling more efficient tumor treatment and monitoring and reducing the treatment burden on patients.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- 亳州优开生物医药科技有限公司
- Filing Date
- 2024-12-23
- Publication Date
- 2026-06-23
AI Technical Summary
Existing photosensitizers have low singlet oxygen quantum yields, resulting in low treatment efficiency and a heavy burden on patients during the treatment process.
A novel photosensitizer compound C is provided, which is prepared by reacting a compound A with a specific structure and a compound B under reflux in acetonitrile, adding acetic acid and pyrrolidine as catalysts, and purifying by column chromatography to produce a photosensitizer with a long near-infrared fluorescence emission wavelength and a high singlet oxygen yield.
Compound C has a longer near-infrared fluorescence emission wavelength, which improves the signal-to-noise ratio, making targeted imaging and lesion site monitoring more sensitive and accurate. Its high singlet oxygen production rate shortens treatment time, reduces patient burden, and improves treatment efficiency.
Smart Images
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Abstract
Description
Technical Field
[0001] This invention relates to the field of biomedical technology, and in particular to a photosensitizer, its preparation method, and its application. Background Technology
[0002] Malignant tumors have become a major threat to human health and life. How to effectively diagnose and treat cancer has been a hot topic of research for scholars.
[0003] Photosensitizers are molecules that can be activated by light of specific wavelengths, thereby producing singlet oxygen or other reactive oxygen species (ROS). ROS have significant cytotoxicity, directly inducing oxidative stress, apoptosis, and necrosis in tumor cells, thus achieving a killing effect on tumor cells. Photodynamic therapy (PDT) offers better selectivity and local control compared to traditional chemotherapy and radiotherapy, reducing damage to normal tissues and effectively avoiding drug resistance. In recent years, in-depth research into the tumor microenvironment has enabled photosensitizers to accumulate more precisely in tumor tissue, enhancing therapeutic efficacy. The application of photosensitizers in tumors has become an important direction in modern tumor treatment research, especially in photodynamic therapy, which shows great potential. Furthermore, photosensitizers also have immunomodulatory effects; by releasing tumor-associated antigens, they can activate the body's immune system, thereby further inhibiting tumor growth both locally and systemically.
[0004] Therefore, the multifunctionality of photosensitizers in tumor treatment not only provides a new approach for precision medicine, but also brings hope for improving patients' survival rate and quality of life. Summary of the Invention
[0005] To address the problem of low singlet oxygen quantum yield in current photosensitizers, this invention provides a photosensitizer, its preparation method, and its application. This invention provides a photosensitizer with the structural formula shown in formula (C), which exhibits excellent properties such as high singlet oxygen quantum yield and a relatively long emission wavelength.
[0006] The specific technical solution of this invention is as follows: This invention provides a photosensitizer, namely compound C, whose structural formula is shown below: , The R base is selected from either H or I.
[0007] The compound C provided by this invention has a longer near-infrared fluorescence emission wavelength, such as 745 nm for compound (4). Longer wavelength light avoids background autofluorescence in the visible light band, thus improving the signal-to-noise ratio, obtaining clearer images, and making detection in tumor tissue more sensitive and accurate, which is helpful for targeted imaging and real-time monitoring of lesions. Simultaneously, compound C has a higher singlet oxygen yield, with an 1O2 yield of up to 78.37%. Photosensitizers with high singlet oxygen yields can achieve the desired killing effect more quickly, thereby shortening irradiation time, reducing the burden on patients during treatment, and improving treatment efficiency.
[0008] As a preferred method, compound C can be any one of the following compounds: This invention provides a method for preparing a photosensitizer, comprising the following steps: Compound A and compound B are reacted to obtain compound C; in: The structural formula of compound A is: The structural formula of compound B is: The structural formula of compound C is:
[0009] As a preferred embodiment of the above preparation method, compound A and compound B are dissolved in acetonitrile for reaction.
[0010] As a preferred embodiment of the above preparation method, a catalyst is added to the reaction.
[0011] More preferably, the catalyst is acetic acid and pyrrolidine.
[0012] As a preferred embodiment of the above preparation method, the reaction is a reflux reaction at 70–90°C.
[0013] As a preferred embodiment of the above preparation method, the reaction time is 4 to 6 hours.
[0014] That is, the reaction process of the above preparation method is as follows: As a preferred embodiment of the above preparation method, the reaction solution obtained from the reaction is processed as follows: extracted with ethyl acetate, the upper layer is collected, concentrated under reduced pressure, purified by column chromatography using a mixed solution of petroleum ether and ethyl acetate as the eluent, the eluent containing the target product is collected, and concentrated under reduced pressure.
[0015] Further preferred, the volume ratio of petroleum ether to ethyl acetate is 4:1.
[0016] In addition to the above, the present invention also provides the application of the above-mentioned compound C / photosensitizer in the preparation of formulations for eliminating cancer cells.
[0017] Compared with the prior art, the present invention has the following technical effects: The compound C provided by this invention has a longer near-infrared fluorescence emission wavelength, such as 745 nm for compound (4). Longer wavelength light avoids background autofluorescence in the visible light band, thus improving the signal-to-noise ratio, obtaining clearer images, and making detection in tumor tissue more sensitive and accurate, which is helpful for targeted imaging and real-time monitoring of lesions. Simultaneously, compound C has a higher singlet oxygen yield, with an 1O2 yield of up to 78.37%. Photosensitizers with high singlet oxygen yields can achieve the desired killing effect more quickly, thereby shortening irradiation time, reducing the burden on patients during treatment, and improving treatment efficiency. Attached Figure Description
[0018] Figure 1 The 1H NMR spectrum of compound (1) prepared in Example 1 of this invention.
[0019] Figure 2 The 1H NMR spectrum of compound (2) prepared in Example 2 of this invention.
[0020] Figure 3 The 1H NMR spectrum of compound (3) prepared in Example 3 of this invention.
[0021] Figure 4 The 1H NMR spectrum of compound (4) prepared in Example 4 of this invention.
[0022] Figure 5 The fluorescence absorption and emission spectra of compounds (1), (2), (3), and (4) in this invention are shown.
[0023] Figure 6 These are the singlet oxygen measurement curves for compounds (1), (2), (3), and (4) in this invention.
[0024] Figure 7 This is a spectrum of the antitumor activity of compound (4) prepared in Example 4 of this invention. Detailed Implementation
[0025] This invention provides a photosensitizer, namely compound C, whose structural formula is shown below: The R base is selected from either H or I.
[0026] This invention also provides a method for preparing a photosensitizer, comprising the following steps: Compound A and compound B are reacted to obtain compound C; in: The structural formula of compound A is: The structural formula of compound B is:
[0027] Compound A is a publicly disclosed compound, and its synthetic method can be found in the following references: Radunz S, Wedepohl S. M, et al. pH-Activatable singlet oxygen-generating boron-dipyrromethenes (BODIPYs) for photodynamic therapy and bioimaging[J]. Journal of MedicinalChemistry, 2020, 63(4): 1699-1708.
[0028] Compound B is a publicly disclosed compound, and its synthesis method can be found in the following reference: Harshini D, Angela VM, Imran PM, et al. Arylacetylene End-Capped Triarylamine-Based D–A System for Switching from Binary to Ternary Memory Behavior[J].ACS Applied Electronic Materials, 2023, 5(11): 6420-6432.
[0029] The present invention will be further described using the following compounds (1) to (4) as examples, in conjunction with the embodiments:
[0030] Those skilled in the art will be able to implement the present invention based on these descriptions. Furthermore, the embodiments of the invention described below are generally only a part of the embodiments of the invention, and not all of them. Therefore, all other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort should fall within the scope of protection of the present invention.
[0031] Example 1 Preparation of compound (1) BDP (50 mg) and 4-diphenylaminobenzaldehyde (48 mg) were dissolved in 10 mL of anhydrous acetonitrile. Acetic acid (24 μL) and tetrahydropyrrole (35 μL) were added to the mixed solution. The reaction was heated under reflux at 80 °C for 4 h, and the reaction progress was monitored by TLC. After the reaction was completed, the mixture was extracted with ethyl acetate, and the supernatant was washed with water and saturated brine. The organic phase was concentrated and separated by column chromatography with petroleum ether / ethyl acetate as the eluent in a 4:1 ratio. The eluent containing the target product (purple eluent) was collected, concentrated under reduced pressure, and dried to obtain an oily liquid product, namely compound (1).
[0032] The obtained product was subjected to NMR analysis, and the results are shown in the figure. Figure 1 .
[0033] In this embodiment, the structural formula of compound (1) is as follows:
[0034] Example 2 Preparation of compound (2) BDP (70 mg) and 4-[bis(4-iodophenyl)amino]benzaldehyde (129 mg) were dissolved in 10 mL of anhydrous acetonitrile. Acetic acid (24 μL) and tetrahydropyrrole (35 μL) were added to the mixed solution. The reaction was heated under reflux at 80 °C for 4 h, and the reaction progress was monitored by TLC. After the reaction was completed, the mixture was extracted with ethyl acetate, and the supernatant was washed with water and saturated brine. The organic phase was concentrated and separated by column chromatography with petroleum ether / ethyl acetate as the eluent in a 5:1 ratio. The eluent containing the target product was collected, concentrated under reduced pressure, and dried to obtain product compound (2).
[0035] The obtained product was subjected to NMR analysis, and the results are shown in the figure. Figure 2 .
[0036] In this embodiment, the structural formula of compound (2) is as follows:
[0037] Example 3 Preparation of compound (3) 2I-BDP (50 mg) and 4-[bis(4-iodophenyl)amino]benzaldehyde (27.7 mg) were dissolved in 10 mL of anhydrous acetonitrile. Acetic acid (24 μL) and tetrahydropyrrole (35 μL) were added to the mixed solution. The reaction was heated under reflux at 80 °C for 4 h, and the reaction progress was monitored by TLC. After the reaction was completed, the mixture was extracted with ethyl acetate, and the supernatant was washed with water and saturated brine. The organic phase was concentrated and separated by column chromatography with petroleum ether / ethyl acetate as the eluent in a 4:1 ratio. The eluent containing the target product was collected, concentrated under reduced pressure, and dried to obtain the product compound (3).
[0038] The obtained product was subjected to NMR analysis, and the results are shown in the figure. Figure 3 .
[0039] In this embodiment, the structural formula of compound (3) is as follows:
[0040] Example 4 Preparation of compound (4) 2I-BDP (50 mg) and 4-diphenylaminobenzaldehyde (53.2 mg) were dissolved in 10 mL of anhydrous acetonitrile. Acetic acid (24 μL) and tetrahydropyrrole (35 μL) were added to the mixed solution. The reaction was heated under reflux at 80 °C for 4 h, and the reaction progress was monitored by TLC. After the reaction was completed, the mixture was extracted with ethyl acetate, and the supernatant was washed with water and saturated brine. The organic phase was concentrated and separated by column chromatography with petroleum ether / ethyl acetate as the eluent at a ratio of 4:1. The eluent containing the target product was collected, concentrated under reduced pressure, and dried to obtain the product compound (4).
[0041] The obtained product was subjected to NMR analysis, and the results are shown in the figure. Figure 4 .
[0042] In this embodiment, the structural formula of compound (4) is as follows:
[0043] Example 5: Fluorescence Spectroscopy Detection of Compounds (1), (2), (3), and (4) A certain amount of compounds (1), (2), (3), and (4) were weighed and prepared into 1 mM probe stock solutions using dimethyl sulfoxide. 2 μL of each stock solution was added to 398 μL of PBS buffer and transferred to a 96-well plate. The absorbance was measured using a UV spectrophotometer. The fluorescence absorption spectra and fluorescence spectroscopy of compounds (1), (2), (3), and (4) were also determined. The experimental results are shown in [Figure number missing]. Figure 5 Among them, compounds 1, 2, 3 and 4 are the experimental results of compounds (1), (2), (3) and (4), respectively.
[0044] from Figure 5 As can be seen, compound (4) has the longest near-infrared fluorescence emission.
[0045] Example 6: Determination of Singlet Oxygen Yield of Compounds (1), (2), (3), and (4) Compounds (1), (2), (3), and (4) were weighed and added to DMSO to prepare a 10 mmol / L stock solution. Then, 10 μL of each stock solution was dissolved in PBS to prepare a 10 μmol / L test solution. SOSG was dissolved in DMSO to prepare a 10 mmol / L stock solution. The SOSG stock solution was diluted to 5 μM with PBS and stored in the dark to prevent premature photodegradation. The prepared SOSG solution was added to the pre-set photosensitizer solution in a 96-well plate. The sample was placed under a light source for irradiation. After irradiation, the fluorescence intensity of the sample was measured immediately. SOSG emits strong green light after detecting singlet oxygen, with an emission wavelength of 530 nm. The fluorescence intensity of compounds (1), (2), (3), and (4) at 530 nm was recorded every 10 seconds after the addition of SOSG. The experimental results are shown in [Figure 1]. Figure 6 Among them, compounds 1, 2, 3 and 4 are the experimental results of compounds (1), (2), (3) and (4), respectively.
[0046] from Figure 6 As can be seen from the data, compound (4) has a high singlet oxygen quantum yield. The 1O2 yield of compound (4) is 78.37%, while the 1O2 yield of the common commercial photosensitizer MB is 52%.
[0047] Example 7: Detection of the antitumor activity of compound (4) The tumor cell line was seeded into 96-well plates, with approximately 5000-10000 cells per well, and cultured for 24 hours to allow adhesion. The cells were incubated at 37°C and 5% CO2. Solutions of compound (4) at different concentrations were prepared, and the photosensitizer was added to each well, ensuring a concentration gradient. Experimental and control groups were set up. The experimental group was used for comparison experiments with and without light, while the control group only had culture medium added without the photosensitizer. The cells were incubated at 37°C for 2 hours to ensure sufficient absorption of the photosensitizer. The group with light was placed in a PDT light source for specific wavelength illumination, while the group without light was kept in darkness. Cell viability was measured using CCK-8 or MTT reagents, and cell survival rate was calculated. The results of the group with and without light were compared. Experimental results are shown below. Figure 7 .
[0048] Depend on Figure 7 The experimental results showed that compound (4) had low toxicity to tumor cells in the dark, but when treated with light, it had a good photodynamic ablation effect on tumor cells due to the photodynamic therapy effect of compound (4), and the photodynamic therapy effect gradually increased with the increase of photosensitizer concentration.
[0049] Unless otherwise specified, the raw materials and equipment used in this invention are all commonly used in the field; unless otherwise specified, the methods used in this invention are all conventional methods in the field.
[0050] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Any simple modifications, alterations, and equivalent transformations made to the above embodiments based on the technical essence of the present invention shall still fall within the protection scope of the present invention.
Claims
1. A photosensitizer, characterized in that: The structural formula is as follows: The R base is selected from either H or I.
2. A method for preparing a photosensitizer, characterized in that: Includes the following steps: Compound A and compound B are reacted to obtain compound C; in: The structural formula of compound A is: The structural formula of compound B is: The structural formula of compound C is:
3. The preparation method according to claim 2, characterized in that: Compound A and compound B are dissolved in acetonitrile to carry out the reaction.
4. The preparation method according to claim 2, characterized in that: A catalyst is added to the reaction.
5. The preparation method according to claim 4, characterized in that: The catalyst is acetic acid and pyrrolidine.
6. The preparation method according to claim 2, characterized in that: The reaction is a reflux reaction at 70–90°C.
7. The preparation method according to claim 6, characterized in that: The reaction takes 4 to 6 hours.
8. The preparation method according to claim 2, characterized in that: The reaction solution obtained from the reaction is processed as follows: extracted with ethyl acetate, the upper layer is collected, concentrated under reduced pressure, purified by column chromatography using a mixed solution of petroleum ether and ethyl acetate as the eluent, the eluent containing the target product is collected, and concentrated under reduced pressure.
9. The preparation method according to claim 8, characterized in that: The volume ratio of petroleum ether to ethyl acetate is 4–5:
1.
10. The use of the photosensitizer as described in claim 1 in the preparation of a formulation for eliminating cancer cells.