A bodipy derivative small molecule and a preparation method and application thereof
The small molecules of BODIPY derivatives prepared by coupling reaction of responsive BODIPY with triphenylamine analogues generate ROS under ultrasound, which solves the problem of poor deep antibacterial effect of photodynamic therapy in the prior art, and achieves deep antibacterial and anti-drug-resistant bacterial effects. Moreover, the preparation method is simple and the product is stable.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- CHANGCHUN UNIV OF SCI & TECH
- Filing Date
- 2023-06-19
- Publication Date
- 2026-07-21
Smart Images
Figure CN116854716B_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of biochemical technology, specifically relating to a small molecule BODIPY derivative, its preparation method, and its application. Background Technology
[0002] Bacteria, as ancient and tiny organisms, have a profound impact on human daily life. Apart from some beneficial probiotics, most bacteria cause infectious diseases and pose a threat to human health. If left uncontrolled, the acute or chronic inflammatory responses they trigger can further lead to irreversible tissue damage. Currently, antibiotics are the ideal treatment strategy for preventing bacterial infections and reducing bacterial virulence. However, the discovery and development of new antibiotics significantly impacts their antibacterial efficacy compared to bacterial evolution and drug resistance. Furthermore, the application of further therapeutic drugs is hindered due to poor specificity against bacteria and the inevitable damage to normal tissues.
[0003] Nanomedicines refer to drugs that are combined with nanoparticles (NPs), nanospheres (NSs), nanocapsules (NCs), etc., as carriers in a certain way. Their particle size may exceed 100 nm, but should usually be less than 500 nm. Nanomedicines can also be nanoparticles made directly from raw drug materials.
[0004] Besides traditional drugs that can be prepared into nanomedicines, nanoparticles prepared by encapsulating small organic molecules with amphiphilic carriers are also widely used in drug research. Since drugs act directly on the human body, they must meet conditions such as non-toxicity, good biocompatibility, and biodegradability. Although various nanomedicines encapsulated with small organic molecules have been disclosed, further research has revealed several shortcomings. For example, most current methods using small molecules containing BODIPY derivatives employ photodynamic therapy via 660 or 808 excitation, but due to the low light penetration within the body, this method is limited to superficial tissues and cannot achieve deep antibacterial effects.
[0005] Because of the aforementioned shortcomings of common BODIPY derivatives, the field has been dedicated to finding alternative methods to achieve deep antibacterial effects, thereby further improving the therapeutic efficacy of small molecule nanomedicines. Summary of the Invention
[0006] The purpose of this invention is to provide a small molecule of BODIPY derivative, its preparation method, and its application. This small molecule is a BODIPY derivative that can generate ROS in response to achieve anti-inflammatory and antibacterial effects in vivo, thereby solving the problem of achieving deep antibacterial effects through ultrasound and replacing broad-spectrum antibiotics to resist drug-resistant bacteria. This further improves the therapeutic effect of small molecule nanomedicines and can replace commercially available broad-spectrum antibiotics.
[0007] The objective of this invention is achieved through the following technical solution:
[0008] In a first aspect, the present invention provides a small molecule of BODIPY derivative, which is obtained by coupling reaction of responsive BODIPY with a triphenylamine analog and can generate ROS under ultrasonic conditions.
[0009] Furthermore, the molar ratio of the responsive BODIPY to the triphenylamine analog is 1:3.
[0010] Furthermore, the responsive BODIPY is selected from one or more of the following compounds:
[0011]
[0012]
[0013] Furthermore, the triphenylamine analogue is selected from one or more of the following compounds:
[0014]
[0015] A second aspect of the present invention provides a method for preparing the small molecules of the BODIPY derivative, comprising the following steps:
[0016] The BODIPY monomer, triphenylamine monomer, Pd2(dba)3, disodium tert-butyl, P(t-Bu)3 and toluene solution were stirred at 85°C under a nitrogen atmosphere for 24 hours; the reaction mixture was then diluted with ethyl acetate and washed with water and saturated NaCl solution; the organic layer was dried on anhydrous Na2SO4, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography.
[0017] A third aspect of the present invention provides the application of the aforementioned BODIPY derivative small molecules in the preparation of antibacterial nanosolutions.
[0018] Furthermore, small molecules of BODIPY derivatives were prepared into nanoparticles by coating them with amphiphilic polymers. These nanoparticles were then injected intravenously and accumulated in the bloodstream at the wound site. ROS were generated by ultrasound to kill bacteria.
[0019] Furthermore, the amphiphilic polymer is PLGA-PEG, DSPE-PEG, or PEG-b-PPG-b-PEG.
[0020] Compared with the prior art, the beneficial effects of the present invention are:
[0021] 1. The small molecules of the BODIPY derivative provided by this invention have the ability to generate reactive oxygen species (ROS) in response. They generate ROS under ultrasound conditions. Compared with other BODIPY derivatives that are treated by photothermal and photodynamic therapy, they have the ability to penetrate deeply and can act on deep bacterial infections or excessive inflammation.
[0022] 2. The small molecules provided by this invention have an ultrasonic effect, and the prepared small molecules have strong absorption in the range of 600-1200nm;
[0023] 3. The small molecules provided by this invention can be encapsulated by an amphiphilic carrier and self-assembled into nanoparticles, thereby improving the stability of the drug;
[0024] 4. The present invention provides a corresponding preparation method with a simple process flow and high product stability. Attached Figure Description
[0025] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.
[0026] Figure 1 1H NMR of small molecules of BODIPY derivatives;
[0027] Figure 2 This is a particle size distribution diagram of the nanoparticles.
[0028] Figure 3 This is a potential diagram of nanoparticles;
[0029] Figure 4 The image shows the UV absorption spectrum of a small molecule of the BODIPY derivative.
[0030] Figure 5 The UV absorption spectrum of nanoparticles encapsulating small molecules of BODIPY derivatives;
[0031] Figure 6 shows the in vitro antibacterial effects against different drug-resistant bacteria;
[0032] Figure 7 shows scanning electron microscope images of bacterial cells after different treatments. Figure 7A Here are scanning electron microscope images of drug-resistant Escherichia coli. Figure 7BScanning electron microscope image of drug-resistant Staphylococcus aureus;
[0033] Figures 8-9 Roadmap for the preparation of several small molecules of BODIPY derivatives;
[0034] Figure 10 Roadmap for the synthesis of compound 1;
[0035] Figure 11 Roadmap for the synthesis of compound 2;
[0036] Figure 12 Roadmap for the synthesis of compound 3. Detailed Implementation
[0037] The present invention will be further described below with reference to embodiments:
[0038] The present invention will now be described in further detail with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and not intended to limit it. Furthermore, it should be noted that, for ease of description, the accompanying drawings show only the parts relevant to the present invention, and not all of the structures.
[0039] It should be noted that similar reference numerals and letters in the following figures indicate similar items; therefore, once an item is defined in one figure, it does not need to be further defined and explained in subsequent figures. Furthermore, in the description of this invention, terms such as "first," "second," etc., are used only to distinguish descriptions and should not be construed as indicating or implying relative importance.
[0040] This invention provides a small molecule of BODIPY derivative, which is obtained by coupling reaction of responsive BODIPY with a triphenylamine analog and can generate ROS under ultrasonic conditions.
[0041] The molar ratio of the responsive BODIPY to the triphenylamine analog is 1:3.
[0042] The responsive BODIPY is selected from one or more of the following compounds:
[0043]
[0044] The triphenylamine analogue is selected from one or more of the following compounds:
[0045]
[0046] Secondly, a method for preparing the aforementioned BODIPY derivative small molecules is provided, comprising the following steps:
[0047] 70 mg (0.1 mmol) of BODIPY monomer, 200.0 mg (0.3 mmol) of triphenylamine monomer, 25 mg (0.03 mmol) of Pd2(dba)3, 15 mg (0.16 mmol) of disodium tert-butyl, and 50 μL of P(t-Bu)3 were stirred with 5 mL of toluene solution at 85 °C under a nitrogen atmosphere for 24 hours. The reaction mixture was then diluted with 50 mL of ethyl acetate and washed with 150 mL of water and 30 mL of saturated NaCl solution. The organic layer was dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography.
[0048] Thirdly, the application of the aforementioned BODIPY derivative small molecules in the preparation of antibacterial nanosolutions is provided.
[0049] Specifically, small molecules of BODIPY derivatives are coated with amphiphilic polymers to prepare nanoparticles, which are then injected intravenously and accumulate in the bloodstream at the wound site. ROS are generated by ultrasound, which ruptures the bacterial cell membrane and kills the bacteria.
[0050] The amphiphilic polymer is PLGA-PEG, DSPE-PEG, or PEG-b-PPG-b-PEG.
[0051] This invention relates to a small molecule of BODIPY derivative, obtained by coupling a BODIPY material to a triphenylamine analog. The overuse and misuse of antibiotics leads to bacterial evolution and resistance. The BODIPY derivative small molecule provided by this invention can be used as a sonosensitive agent, replacing antibiotics to achieve antibacterial effects without causing bacterial resistance. This BODIPY derivative small molecule exhibits good antibacterial effects against both common and drug-resistant bacteria. Furthermore, this small molecule can rapidly generate ROS upon ultrasound, which can be used in sonodynamic therapy, photodynamic therapy, etc.
[0052] Example 1 Monomer 1
[0053] 2,4-Dimethylpyrrole (10 g, 1 mmol), trifluoroacetic acid (5.75 g, 0.48 mmol), phenyltrichlorosilane (10.67 g, 0.48 mmol), NEt3 (15 mL, 1.05 mmol), and boron trifluoride diethyl ether (75 mL, 1.65 mmol) were dissolved in 200 mL of CH2Cl2 and reacted at 0°C for 3 h. After the reaction was completed, 100 mL of H2O was added, and the mixture was extracted three times with 100 mL of dichloromethane. The mixture was separated by column chromatography to give a purple solid with a yield of 73%.
[0054] Example 2 Small Molecule
[0055] Fluoroboropyrrole (BODIPY) (250 mg, 1 mmol), 4-bromobenzaldehyde (150 mg, 2.5 mmol), β-alanine (12 mg, 1 mmol), and piperidine (120 μL) were dissolved in 30 mL of anhydrous toluene and reacted at 80°C for 24 h. The mixture was separated by column chromatography to give a dark green solid, as shown in Formula 2, with a yield of 68%. 1 H NMR (400MHz, Chloroform-d 7.20 -6.94 (m, 15H), 6.84 (s, 4H), 6.67 (s, 1H).
[0056] Example 3: Synthesis of Compound 1
[0057] The small molecule (70 mg, 0.1 mmol) from Example 2 was stirred for 24 h at 85 °C under a nitrogen atmosphere with a solution of triphenylamine (200.0 mg, 0.3 mmol), Pd2(dba)3 (25 mg, 0.03 mmol), disodium tert-butyl (15 mg, 0.16 mmol), P(t-Bu)3 (50 μL), and toluene (5 mL). The reaction mixture was then diluted with ethyl acetate (50 mL) and washed with water (150 mL) and saturated NaCl solution (30 mL). The organic layer was dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to give compound 1 (57.7 mg, yield 46.9%) as a black solid. 1 The H NMR (400MHz, deuterated DMSO) spectrum is shown below. Figure 10 As shown.
[0058] Example 4: Synthesis of Compound 2
[0059] The small molecule (50.0 mg, 0.1 mmol) from Example 2 was mixed with a triphenylamine analog (237.0 mg, 0.3 mmol), Pd2(dba)3 (15 mg, 0.03 mmol), disodium tert-butyl (20 mg, 0.16 mmol), P(t-Bu)3 (35 μL), and a toluene (5 mL) solution and stirred at 110 °C under a nitrogen atmosphere for 48 h. The reaction mixture was then diluted with ethyl acetate (50 mL) and washed with water (150 mL) and a saturated NaCl solution (30 mL). The organic layer was dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to give compound 2 (38.3 mg, yield 76.6%) as a gray powder. 1 The ¹H NMR spectrum (400MHz Chlorofomm-d) is shown below: 7.41 (s, 4H), 7.18–7.99 (m, 44H), 6.99–6.73 (m, 18H), 1.95–2.29 (s, 2H). Figure 11 As shown.
[0060] Example 5: Synthesis of Compound 3
[0061] The small molecule (100 mg, 0.1 mmol) from Example 2 was stirred for 48 h at 110 °C under a nitrogen atmosphere with a triphenylamine analogue (345.0 mg, 0.3 mmol) of Formula 16, Pd2(dba)3 (30 mg, 0.03 mmol), disodium tert-butyl (20 mg, 0.16 mmol), P(t-Bu)3 (65 μL), and toluene (10 mL). The reaction mixture was then diluted with ethyl acetate (100 mL) and washed with water (300 mL) and saturated NaCl solution (50 mL). The organic layer was dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to give compound 3 (76 mg, 76% yield) as a black solid. 1 ¹H NMR (400MHz, DMSO-d6): δppm 7.64-7.60 (d, 4H), 7.48-7.46 (d, 2H), 7.39-7.32 (q, 6H), 7.24 (s, 8H), 7.11-7.09 (t, 10H), 6.98-6.96 (d, 4H), 6.87-6.85 (d, 4H), 2.31 (s, 2H). Spectra are shown below. Figure 12 As shown.
[0062] Example 6 Preparation of Nanoparticles
[0063] 10 mg of compound 1BODIPY (prepared in Example 3) and 100 mg of the amphiphilic polymer PLGA-PEG were weighed and placed in a 25 mL vial containing 10 mL of THF. The mixture was sonicated until both components were completely dissolved, rapidly evaporated to dryness, and 5 mL of water was added dropwise under sonication. Dialysis was performed for 12 h using a dialysis bag with a molecular weight cutoff of 3500. The dialysate was centrifuged at 3000 rpm for 3 min, and the precipitate was discarded to obtain a nanomicelle system. The particle size was determined using dynamic light scattering (DLS), and the results are as follows: Figure 2 As shown, its average particle size (z-average) is 113 nm, its polymer dispersity index (PDI) is 0.21, and its potential is negative. Figure 3 As shown. The ultraviolet absorption of the small molecule of compound 1 was measured using a UV-Vis spectrophotometer (UV-2450PC, Shimazu), and the results are shown below. Figure 4 As shown in the figure, the small molecules exhibit strong absorption in the range of 400-1200.
[0064] The ultraviolet absorption was measured using a UV-2450PC spectrophotometer (Shimazu), and the results are as follows: Figure 5 As shown, by Figure 5 It can be seen that the loaded nanomicelles have strong absorption in the range of 400-1200 nm.
[0065] Example 7 Antibacterial Performance Test
[0066] Dilute drug-resistant Staphylococcus aureus or drug-resistant Escherichia coli in LB liquid medium to OD100 during the logarithmic growth phase. 600 =1.0 colony-forming units (CFU) were used as the working suspension. The volume was 100 μL, with 100 μL of different groups of nanoparticles added (diluted in PBS buffer), followed by 300 μL of PBS, for a final volume of 500 μL. Incubation was performed on a shaker at 37°C for 4 hours. For groups requiring sonication, a 1.5 W cm⁻¹ power was applied after 4 hours. -2 The bacteria were sonicated at 3 MHz for 1 min, and then returned to a shaker for 1 h of incubation. 10 μL of each of the different treatment groups was diluted 10,000 times with PBS. 100 μL of the diluted bacterial solution was spread onto a solid culture medium and placed in an incubator. After 12 h of incubation, the colony count was performed. Figure 6 shows the growth of different bacteria (A is drug-resistant Escherichia coli, B is drug-resistant Staphylococcus aureus) on a solid culture medium after different treatments. It can be seen that the antibacterial effect of the BODIPY molecule of compound 1 increased under ultrasound, indicating that ultrasound action on the OH· produced by the BODIPY molecule disrupts the bacterial cell membrane.
[0067] The concentration of the solid culture medium in which bacterial survival rate is 0 was established as the minimum inhibitory concentration (MIC). After MIC determination, 5 μL of the mixture with no visible bacterial growth was spread onto LB agar plates, which were incubated at 37°C. After 18 hours, bacterial activity was visually observed. Each experiment was repeated three times. The number of E. coli and S. aureus colonies on the agar plates was counted using the plate counting method, and the bacterial survival rate of different treatment groups was calculated according to the following formula:
[0068] Bacterial survival rate (%) = (Number of bacterial colonies in experimental group / Number of bacterial colonies in control group) × 100%
[0069] The antibacterial properties of compound 1BODIPY (small molecule) in Example 3 are shown in Table 1:
[0070] Table 1
[0071]
[0072] Bacterial morphology after treatment in Example 6
[0073] Dilute the bacterial culture to an appropriate concentration (1×10⁻⁶). 6 ~1×10 7 CFU mL -1Different groups of nanoparticles were added to PBS, PBS+US, NP, and NP+US and incubated on a shaker at 37°C for 4 hours. For groups requiring sonication, an ultrasound treatment was administered at a power of 1.5 W / cm² after 4 hours. 2 Sonication for 1 min. After washing the bacterial cells three times with PBS, they were successively treated with 30%, 50%, 70%, 80%, 90%, 95%, and 100% ethanol for 10 min each. Finally, the samples were dried in a vacuum freeze-drying oven and photographed using a scanning electron microscope. As shown in Figure 7, it can be seen that under the action of ultrasound, the surfaces of both bacterial groups showed wrinkling and depressions compared to the PBS group. Furthermore, the treatment with small molecules of compound 1BODIPY from Example 3, encapsulated into nanoparticles and then subjected to ultrasound, caused the bacterial cell membrane to rupture. When the bacterial cell membrane ruptured, nutrients were lost, ultimately leading to bacterial death.
[0074] Note that the above description is merely a preferred embodiment of the present invention and the technical principles employed. Those skilled in the art will understand that the present invention is not limited to the specific embodiments described herein, and various obvious changes, readjustments, and substitutions can be made without departing from the scope of protection of the present invention. Therefore, although the present invention has been described in detail through the above embodiments, the present invention is not limited to the above embodiments, and may include many other equivalent embodiments without departing from the concept of the present invention, the scope of which is determined by the scope of the appended claims.
Claims
1. A small molecule of a BODIPY derivative, characterized in that, Its structure is as follows: ； ； ； ； ； ； ； ； 。 2. The application of a BODIPY derivative small molecule according to claim 1 in the preparation of antibacterial nanosolutions.