A bodipy dye with a cyclic structure and preparation method and application thereof
By designing a cyclic BODIPY dye, the instability in aqueous media and the shortcomings of traditional methods have been overcome, achieving stability and safety in vivo and expanding the application scope of photodynamic and photothermal therapy.
Patent Information
- Application Number
- CN202411634705.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-15
- Publication Date
- 2026-02-03
- Estimated Expiration
- 2044-11-15
AI Technical Summary
Existing BODIPY dyes are unstable in aqueous media, which limits their biomedical applications. Furthermore, traditional preparation methods suffer from problems such as dye leakage, carrier toxicity, and performance instability.
We designed BODIPY dyes with cyclic structures to form amphiphilic nanoparticles through ester or ether bonds, thereby improving their self-assembly stability in aqueous solutions and applying them to integrated tumor diagnosis and treatment research using photodynamic and photothermal therapies.
This enhances the stability and safety of BODIPY dyes in vivo, enables integrated tumor diagnosis and treatment, reduces the requirements for excitation light sources, and expands the scope of clinical applications.
Smart Images
Figure QLYQS_1 
Figure QLYQS_2 
Figure QLYQS_3
Abstract
Description
Technical Field
[0001] This invention relates to the field of medical dye technology, specifically to a BODIPY dye with a cyclic structure, its preparation method, and its application. Background Technology
[0002] Bodipy dyes, short for boron-dipyrromethene, are a class of organic dyes with unique optical properties. Their molecular structure consists of two pyrrole rings connected by a four-membered boron bridge. Due to their excellent optical properties, such as high quantum yield, good photostability, broad absorption bands, and narrow emission bands, bodipy dyes have found wide applications in fields such as biolabeling, photodynamic therapy (PDT), photothermal therapy (PTT), and fluorescence imaging.
[0003] However, most BODIPY dyes are hydrophobic and insoluble in biological media, which hinders their biomedical applications to some extent. One approach to address this issue is to prepare highly stable BODIPY aqueous dispersion nanoparticles (BODIPY NPs). Traditional methods for preparing fluorescent nanoparticles typically involve physically encapsulating or chemically bonding the dye into a polymer bulk or other nanoparticles. However, these methods have several drawbacks. For example, dye leakage from the nanoparticles over time may lead to insufficient effective dye concentrations at tumor sites, thus reducing the efficacy of photodynamic therapy (PDT); the potential toxicity of the carrier may damage normal tissues, increasing side effects during treatment; or batch-to-batch variability due to complex preparation processes may result in unstable performance, affecting the consistency and predictability of treatment outcomes.
[0004] Therefore, in the study of BODIPY dyes as photosensitizers for integrated diagnosis and treatment of tumors, the stability of BODIPY dyes in aqueous media and their safety in vivo are key aspects of this research. Summary of the Invention
[0005] To improve the stability of BODIPY dyes in aqueous media and their safety in organisms, this invention proposes a BODIPY dye with a cyclic structure, its preparation method, and its applications.
[0006] The technical solution of the present invention is as follows:
[0007] A BODIPY dye with a cyclic structure has the following structural formula:
[0008]
[0009] Wherein, R1 is selected from H or CH3; R2 is selected from H, I or Br; R3 is selected from H, CH3 or C6H5; R4 is selected from
[0010] The present invention also provides a method for preparing a BODIPY dye with a cyclic structure, wherein compound A and dicarboxylic acid are dissolved in anhydrous dichloromethane, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and a catalyst are added, and the reaction is carried out under nitrogen protection. The crude product is purified by silica gel column chromatography to obtain a BODIPY dye with a cyclic structure.
[0011] The structural formula of compound A is as follows:
[0012]
[0013] Preferably, the dicarboxylic acid is selected from HOOC(C4H8)COOH, HOOCCH2S-SCH2COOH, HOOCCH2PhCH2COOH, or HOOCCH2(OCH2CH2). n OCH2COOH;
[0014] Among them is HOOCCH2 (OCH2CH2). n OCH2COOH is selected from polyethylene glycol derivatives of different molecular weights, including 200, 400, 600, 800, and 1000 Daltons.
[0015] Preferably, the catalyst is 4-dimethylaminopyridine.
[0016] Preferably, the molar ratio of compound A, dicarboxylic acid and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is 0.123:0.148:0.37.
[0017] Preferably, the reaction is carried out at room temperature for 24 hours.
[0018] The present invention also provides an application of the above-mentioned BODIPY dye with a cyclic structure, specifically: as a photosensitizer applied in the integrated research of tumor diagnosis and treatment using photodynamic and photothermal therapy.
[0019] Compared with the prior art, the specific beneficial effects of the present invention are as follows:
[0020] This invention utilizes cyclic BODIPY formed through ester or ether bonds. This increased amphiphilicity allows for easy self-assembly into nanoparticles in aqueous solution with high stability. This enables in vivo delivery, enhances stability in water, prolongs in vivo circulation time, and increases biocompatibility and safety, thus facilitating integrated in vivo tumor diagnosis and treatment.
[0021] The BODIPY dyes provided by this invention have a simple and well-defined structure, a straightforward synthesis method, and low production costs. BODIPY-based photosensitizers exhibit a high molar extinction coefficient and the advantage of tunable spectra. Their strong absorption in the visible light range reduces the requirements for excitation light sources, which is beneficial for expanding the clinical application of BODIPY photosensitizers and bringing greater social and economic benefits. Attached Figure Description
[0022] Figure 1 The hydrogen nuclear magnetic resonance spectrum of BODIPY-1;
[0023] Figure 2 Electrospray ionization mass spectrometry for BODIPY-1;
[0024] Figure 3 The hydrogen nuclear magnetic resonance spectrum of BODIPY-2;
[0025] Figure 4 The 1H NMR spectrum of BODIPY-4;
[0026] Figure 5 Matrix-assisted laser desorption / ion-time mass spectrometry for BODIPY-4;
[0027] Figure 6 The graphs show the toxicity of BODIPY-2 nanoparticles to mouse breast cancer cells (4T1) under dark and light conditions (C, D) and to normal mouse fibroblasts (L929) under dark and light conditions (A, B).
[0028] Figure 7 The graph shows the toxicity of BODIPY-3 nanoparticles to mouse breast cancer cells (4T1) under dark and light conditions.
[0029] Figure 8 The graph shows the toxicity of BODIPY-4 nanoparticles to mouse breast cancer cells (4T1) under dark and light conditions. Detailed Implementation
[0030] To make the technical solutions of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. It should be noted that the following embodiments are only used to better understand the technical solutions of the present invention and should not be construed as limiting the present invention.
[0031] Example 1.
[0032] In this embodiment, compound A was synthesized through the following steps:
[0033] BODIPY (200 mg, 0.25 mmol) and p-hydroxybenzaldehyde (102 mg, 0.75 mmol) were dissolved in 30 mL of anhydrous DMF, and acetic acid (100 μL) and piperidine (200 μL) were added. The mixture was reacted under nitrogen protection at 80 °C for 20 min. The crude product was purified by silica gel column chromatography to obtain product (compound A).
[0034] Its synthetic route is as follows:
[0035]
[0036] Example 2.
[0037] In this embodiment, compound A prepared in Example 1 is reacted with adipic acid in anhydrous dichloromethane to generate BODIPY-1.
[0038] The specific preparation process is as follows:
[0039] Compound A (100 mg, 0.123 mmol) and HOOC(C4H8)COOH (20 mg, 0.148 mmol) were dissolved in 50 mL of anhydrous dichloromethane. 1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl, 58 mg, 0.37 mmol) and 4-dimethylaminopyridine (DMAP, catalytic amount) were added. The reaction was carried out under nitrogen protection at room temperature for 24 h. The crude product was purified by silica gel column chromatography to obtain product (BODIPY-1).
[0040] Its synthetic route is as follows:
[0041]
[0042] The proton NMR spectrum of BODIPY-1 is shown below. Figure 1 The results of electrospray ionization mass spectrometry are shown in […]. Figure 2 This can prove the structure and purity of BODIPY-1.
[0043] Example 3.
[0044] In this embodiment, compound A prepared in Example 1 is reacted with dicarboxylic acid HOOCCH2S-SCH2COOH in anhydrous dichloromethane to generate BODIPY-2.
[0045] The specific preparation process is as follows:
[0046] Compound A (100 mg, 0.123 mmol) and HOOCCH2S-SCH2COOH (20 mg, 0.148 mmol) were dissolved in 50 mL of anhydrous dichloromethane. 1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl, 58 mg, 0.37 mmol) and 4-dimethylaminopyridine (DMAP, catalytic amount) were added. The reaction was carried out under an inert atmosphere at room temperature for 24 h. The crude product was purified by silica gel column chromatography to obtain product (BODIPY-2).
[0047] Its synthetic route is as follows:
[0048]
[0049] The proton NMR spectrum of BODIPY-2 is shown below. Figure 3 This can prove the structure of BODIPY-2.
[0050] Example 4.
[0051] In this embodiment, compound A prepared in Example 1 is reacted with dicarboxylic acid HOOCCH2PhCH2COOH in anhydrous dichloromethane to generate BODIPY-3.
[0052] The specific preparation process is as follows:
[0053] Compound A (100 mg, 0.103 mmol) and HOOCCH2PhCH2COOH (32 mg, 0.165 mmol) were dissolved in 50 mL of anhydrous dichloromethane. 1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl, 60 mg, 0.38 mmol) and 4-dimethylaminopyridine (DMAP, catalytic amount) were added. The reaction was carried out under an inert atmosphere at room temperature for 24 h. The crude product was purified by silica gel column chromatography to obtain product (BODIPY-3).
[0054] Its synthetic route is as follows:
[0055]
[0056] Example 5.
[0057] This embodiment uses compound A prepared in Example 1 and dicarboxylic acid HOOCCH2(OCH2CH2). n OCH2COOH reacts in anhydrous dichloromethane to produce BODIPY-4.
[0058] The specific preparation process is as follows:
[0059] Compound A (40.6 mg, 0.05 mmol) and HOOCCH2 (OCH2CH2) were mixed.n OCH₂COOH (20 mg, 0.05 mmol) was dissolved in 50 mL of anhydrous dichloromethane, and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl, 48 mg, 0.25 mmol) and 4-dimethylaminopyridine (DMAP, catalytic amount) were added. The reaction was carried out under an inert gas atmosphere at room temperature for 48 h. After the reaction was completed, the product (BODIPY-4) was purified by column chromatography.
[0060] Its synthetic route is as follows:
[0061]
[0062] The proton NMR spectrum of BODIPY-4 is shown below. Figure 4 Matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF-MS) is Figure 5 This can prove the structure and purity of BODIPY-4.
[0063] Example of results.
[0064] The antitumor activity of cyclic BODIPY-2 was evaluated using the MTT assay.
[0065] Mouse breast cancer cells (4T1 cells) and fibroblasts (L929 cells) were seeded into 96-well plates and cultured until cell adhesion was achieved. Different concentrations of BODIPY-2 nanoparticles (0–20 μM) were added to the culture dishes, and after incubation for 4 h, each well was illuminated for 5 min, followed by 24 h of further culture. Then, MTT (20 ml / well) was added. After 4 h, the culture medium was removed from each well, and DMSO (150 μL / well) was added. Finally, the plates were shaken for 3 min, and the relative cell viability was calculated by measuring the absorbance at 492 nm using an enzyme-labeled assay. Each experiment was repeated at least three times.
[0066] The experimental results are shown in Figure 6 The results showed that BODIPY-1 and BODIPY-2 exhibited good biocompatibility under no-light conditions; under light conditions, a low concentration of BODIPY-2 (10 μM) was sufficient to eliminate tumor cells without damaging normal cells.
[0067] The same procedure was used to evaluate the antitumor activity of cyclic BODIPY-3 and BODIPY-4, such as... Figure 7 The cytotoxicity of BODIPY-3 under light and no light conditions was investigated. The results showed that it was essentially non-toxic under no light conditions, while under light conditions, it could kill tumor cells at 20 μM. Figure 8 The cytotoxicity of BODIPY-4 under light and no light conditions was investigated. The results showed that it was essentially non-toxic under no light conditions, while under light conditions, it could kill tumor cells at 60 nM.
[0068] It is understood that the present invention has been described through some embodiments, and those skilled in the art will recognize that various changes or equivalent substitutions can be made to these features and embodiments without departing from the spirit and scope of the invention. Furthermore, under the teachings of the present invention, these features and embodiments can be modified to adapt to specific situations and materials without departing from the spirit and scope of the invention. Therefore, the present invention is not limited to the specific embodiments disclosed herein, and all embodiments falling within the scope of the claims of this application are within the protection scope of the present invention.
Claims
1. A BODIPY dye having a cyclic structure, characterized in that, The structural formula is as follows: ， Wherein, R1 is selected from H or CH3; R2 is selected from H, I or Br; R3 is C6H5; R4 is selected from , or .
2. A method for preparing a BODIPY dye with a cyclic structure as described in claim 1, characterized in that, Compound A and dicarboxylic acid were dissolved in anhydrous dichloromethane, and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and a catalyst were added. The reaction was carried out under nitrogen protection. The crude product was purified by silica gel column chromatography to obtain BODIPY dye with a cyclic structure. The structural formula of compound A is as follows: 。 3. The method for preparing a BODIPY dye with a cyclic structure according to claim 2, characterized in that, The dicarboxylic acid is selected from HOOC(C4H8)COOH, HOOCCH2S-SCH2COOH, or HOOCCH2PhCH2COOH.
4. The method for preparing a BODIPY dye with a cyclic structure according to claim 2, characterized in that, The catalyst is 4-dimethylaminopyridine.
5. The method for preparing a BODIPY dye with a cyclic structure according to claim 2, characterized in that, The molar ratio of compound A, dicarboxylic acid, and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is 0.123:0.148:0.
37.
6. The method for preparing a BODIPY dye with a cyclic structure according to claim 2, characterized in that, The reaction was carried out at room temperature for 24 hours.
7. The application of a BODIPY dye with a cyclic structure as described in claim 1, or a BODIPY dye with a cyclic structure prepared by any one of claims 2 to 6, characterized in that, As a photosensitizer, it is used in the preparation of drugs for the integrated diagnosis and treatment of tumors using photodynamic and photothermal therapy.