Water-soluble sound-sensitive agent nanogel as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of nano biomedical materials, and particularly relates to a water-soluble sound-sensitive agent nanogel and a preparation method and application thereof, the nanogel has the following structural formula I: # imgabs0 #, R is H or 5, 10, 15, 20-tetra (pentafluorophenyl) porphyrin, at least one R is 5, 10, 15, 20-tetra (pentafluorophenyl) porphyrin, and n is a positive integer. Compared with the prior art, the problems that in the prior art, a sound-sensitive agent is poor in water solubility, low in sound-sensitive efficiency and difficult to apply practically are solved, on one hand, the water solubility of the sound-sensitive agent is improved, the biocompatibility of the sound-sensitive agent is improved, on the other hand, the large-size cavity provides a cavitation nucleus for the sound-sensitive agent, and the sonodynamic efficiency is enhanced.

Description

Technical Field

[0001] The invention belongs to the technical field of nano-biomedical materials, and specifically relates to a water-soluble sonosensitizer nanogel and a preparation method and application thereof. Background Art

[0002] Cancer is one of the greatest hazards to human health. Currently, clinical methods for cancer treatment include surgery, chemotherapy, radiotherapy, and immunotherapy. Although these strategies are the current standard treatment options, they also have certain side effects. In recent years, with the vigorous development of nanomedicine, a series of new non-invasive anti-cancer strategies have emerged in order to improve the efficacy and reduce adverse side effects. At the same time, a variety of new cancer treatment methods have been developed in recent years, such as photodynamic therapy, photothermal therapy, and sonodynamic therapy. However, the inherent disadvantages of light in photodynamic therapy, such as shallow tissue penetration depth and skin phototoxicity, including near-infrared (NIR) light, also hinder its development in the treatment of deep tumors. In contrast, ultrasound (US) is a highly penetrating mechanical wave, and sonodynamic therapy (SDT) developed based on this is a non-invasive treatment method based on US. It activates sonosensitizers in response to ultrasound stimulation, producing therapeutic effects such as ROS, cavitation, and hyperthermia. Sonodynamic therapy not only has a penetration ability of up to tens of centimeters in tissues, but also minimizes damage to surrounding normal tissues. Due to these ideal characteristics, ultrasound therapy is particularly suitable for the treatment of cancer. Compared with traditional photodynamic therapy, sonodynamic therapy has obvious advantages as a non-invasive tumor treatment method.

[0003] In 1989, Umemura et al. first discovered that hematoporphyrin has high cytotoxicity under ultrasound irradiation and can be used to kill cancer cells, and named this method sonodynamic therapy (SDT). Since then, SDT has rapidly developed for various biomedical applications. The sonosensitizers used in sonodynamic therapy were porphyrin sonosensitizers. Although many scholars have discovered that other drugs have sonosensitivity, the most commonly used sonosensitizers are still porphyrin compounds, such as hematoporphyrin monomethyl ether (HMME), protoporphyrin (PpIX), dihydrochlorin (Ce6), etc. However, these porphyrin sonosensitizers usually have problems such as low water solubility, rapid metabolism, poor bioavailability and stability.

[0004] Therefore, the research and development of modified new porphyrin-based sonosensitizers has always been a hot topic in the field of sonodynamic therapy. Summary of the invention

[0005] The purpose of the present invention is to provide a water-soluble sonosensitizer nanogel and its preparation method and application in order to solve at least one of the above problems, so as to solve the problems of poor water solubility, low sonosensitivity efficiency and difficulty in practical application of sonosensitizers in the prior art. On the one hand, this solution improves the water solubility of the sonosensitizer and enhances the biocompatibility of the sonosensitizer. On the other hand, the large-sized cavity provides a cavitation nucleus for the sonosensitizer and enhances the sonodynamic efficiency.

[0006] The purpose of the present invention is achieved through the following technical solutions:

[0007] The first aspect of the present invention discloses a water-soluble sonosensitizer nanogel (CD-TFPP), wherein the nanogel has the structural formula of the following formula I:

[0008]

[0009] Wherein, R is H or 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin and at least one R is 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin, and n is a positive integer.

[0010] Preferably, the size of the nanogel is 35 to 110 nm.

[0011] More preferably, the size of the nanogel is 35 nm, 75 nm or 110 nm.

[0012] The second aspect of the present invention discloses a method for preparing the water-soluble sonosensitizer nanogel as described above, wherein the nanogel is prepared by a nucleophilic substitution reaction of a cyclic compound β-cyclodextrin (β-CD) having a large cavity and a sonosensitizer 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin (TFPP) having a sonosensitizing effect.

[0013] Preferably, the method comprises the following steps:

[0014] S1: in a first solvent, β-cyclodextrin and 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin react under the condition of a catalyst, and then dialyze and freeze-dry to obtain a reaction product;

[0015] S2: dissolving the reaction product solvent obtained in step S1 in a second solvent to form an organic phase, and dripping the organic phase into the water phase stirred at a constant temperature until the mixed solution is stable;

[0016] S3: filtering the mixed solution obtained in step S3 through an aqueous phase membrane to obtain the nanogel.

[0017] Preferably, in step S1, the first solvent includes N-methylpyrrolidone, the catalyst includes anhydrous potassium carbonate; and the molar ratio of β-cyclodextrin to 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin is 1:0.5-1.5.

[0018] More preferably, the molar ratio of β-cyclodextrin to 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin is 1:0.5, 1:1 or 1:1.5.

[0019] Preferably, in step S1, the reaction temperature is 80-90°C and the reaction time is 35-50 hours; the dialysis is performed in ultrapure water using a dialysis bag with a molecular weight cutoff of 8000-14000, and the dialysis time is 48-60 hours.

[0020] Preferably, in step S2, the second solvent includes dimethyl sulfoxide; and the volume ratio of the organic phase to the aqueous phase is 1:3-5.

[0021] More preferably, the volume ratio of the organic phase to the aqueous phase is 1:4.

[0022] Preferably, in step S2, the temperature of the aqueous phase is 20-30°C.

[0023] Preferably, in step S3, the aqueous phase membrane is an aqueous phase membrane of 200 to 300 nm.

[0024] The third aspect of the present invention discloses a use of the water-soluble sonosensitizer nanogel as described above in the preparation of a sonosensitizer for sonodynamic therapy.

[0025] Compared with the prior art, the present invention has the following advantages:

[0026] The present invention modifies the sonosensitizer 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin (TFPP) with sonosensitizing effect. The perfluoroporphyrin has a clear structure and high photostability, and the introduced macrocyclic main body β-CD improves the water solubility and biocompatibility, thereby obtaining a new water-soluble sonosensitizer with good biosafety and good sonodynamic effect, overcoming the shortcomings of easy aggregation and low bioavailability of traditional small molecule sonosensitizers, and significantly improving the SDT efficacy.

[0027] Compared with traditional sonosensitizers, the porphyrin sonosensitizer nanogel of the present invention has a simple preparation method, high batch stability, good biocompatibility, low toxic side effects, significant sonotoxicity, etc. In addition, the multiple main molecule cyclodextrins carried by the nanogel can form a host-guest complex with a variety of guest small molecule drugs for combined therapy, and can easily achieve functional customization, providing a platform for personalized treatment, and has potential applications in cancer treatment.

[0028] Therefore, the nanogel sonosensitizer has good application prospects in tumor treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 is the UV-visible absorption spectra of CD-TFPP-1, CD-TFPP-2 and CD-TFPP-3 in aqueous solution in Example 1;

[0030] Figure 2 is the fluorescence emission spectra of CD-TFPP-1, CD-TFPP-2 and CD-TFPP-3 in aqueous solution in Example 2;

[0031] Figure 3 The particle size distribution diagram of CD-TFPP-1, CD-TFPP-2 and CD-TFPP-3 in Example 3;

[0032] Figure 4 The UV-visible absorption spectra of CD-TFPP-1, CD-TFPP-2 and CD-TFPP-3 under continuous ultrasound and the line graph of the acoustic stability of the three nanoparticles in Example 4;

[0033] Figure 5 The water produced under ultrasonic conditions in Example 5 1 Effect of O2 on UV-visible light absorption of scavenger DPBF;

[0034] Figure 6 The CD-TFPP-1 produced under ultrasonic conditions in Example 5 1 Effect of O2 on UV-visible light absorption of scavenger DPBF;

[0035] Figure 7 The CD-TFPP-2 produced under ultrasonic conditions in Example 5 1 Effect of O2 on UV-visible light absorption of scavenger DPBF;

[0036] Figure 8 The CD-TFPP-3 produced under ultrasonic conditions in Example 5 1 Effect of O2 on UV-visible light absorption of scavenger DPBF;

[0037] Fig. 9 The water and CD-TFPP-1, CD-TFPP-2, CD-TFPP-3 produced under ultrasonic conditions in Example 5 1 Line graph of the absorbance change of O2 to capture agent DPBF;

[0038] Fig.10 This is a line graph of the size change of CD-TFPP in Example 6 after being left for several days;

[0039] Fig.11 This is the confocal imaging image (50 μm) of cell uptake after co-incubation of CD-TFPP and 4T1 cells in Example 7;

[0040] Fig.12 The cell activity of CD-TFPP after co-incubation with L929 determined by the MTT method in Example 8;

[0041] Fig.13 The dark cytotoxicity of CD-TFPP in different cell lines determined by the MTT method in Example 8;

[0042] Fig.14 The cell sonotoxicity of CD-TFPP in different cell lines determined by the MTT method in Example 8;

[0043] Fig.15 These are confocal fluorescence images (300 μm) of 4T1 cells co-stained with calcein AM and propidium iodide PI after different treatments in Example 9. DETAILED DESCRIPTION

[0044] The present invention is described in detail below with reference to the accompanying drawings and specific embodiments.

[0045] Unless otherwise specified in the following description, the reagents used are conventional commercially available products, and the methods used are well known in the art.

[0046] A modifiable water-soluble sonosensitizer CD-TFPP is specifically obtained by reacting a widely used water-soluble macrocyclic main body β-CD and a hydrophobic photosensitizer drug TFPP with sonosensitizing effect. β-CD, TFPP and CD-TFPP have the following structural formulas respectively:

[0047]

[0048] The preparation method of sonosensitizer nanogel comprises: reacting beta-cyclodextrin and 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin in a certain proportion, using anhydrous potassium carbonate (K2CO3) as a catalyst, using N-methylpyrrolidone (NMP) as a solvent, reacting at a temperature of 80-90°C, reacting for 35-50 hours, dialyzing in ultrapure water after the reaction and freeze-drying to obtain a reddish brown product; dissolving the reddish brown product in dimethyl sulfoxide to make it dissolve evenly, slowly dripping the evenly mixed organic phase solution into a water phase with a high-speed stirring constant temperature of 20-30°C by using a syringe, the organic phase to water phase (volume ratio) being 1:3-5, and continuing stirring until the mixed solution is stable; passing the stirred mixed solution through a 200-300 nm water phase membrane to obtain a clear CD-TFPP nanogel solution (CD-TFPP).

[0049] The synthesis route is shown in the following reaction formula, taking the reaction at 85°C for 40h as an example:

[0050]

[0051] in:

[0052] The feed ratio (molar ratio) of β-cyclodextrin and 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin is β-CD:TFPP=1:1.5-0.5, specifically 1:0.5, 1:1 or 1:1.5.

[0053] The dialysis process uses a dialysis bag with a molecular weight cutoff of 8000-14000, and the dialysis time is 48h-60h.

[0054] The sonosensitizer nanogel provided by the present invention is subjected to photophysical property analysis, and the experimental results show that the sonosensitizer nanogel CD-TFPP has good stability.

[0055] The sonosensitizer nanogel provided by the present invention is subjected to singlet oxygen generation performance analysis, and experimental results show that the sonosensitizer nanogel CD-TFPP has a high efficiency in singlet oxygen generation capability.

[0056] The sonosensitizer nanogel provided by the present invention is subjected to an in vitro cell biocompatibility experiment, and the experimental results show that the sonosensitizer nanogel CD-TFPP has almost no toxic effect on normal cells, and its bioavailability is greatly improved compared with the unmodified sonosensitizer TFPP.

[0057] The sonosensitizer nanogel provided by the present invention is subjected to an in vitro cell uptake experiment, and the experimental results show that the sonosensitizer nanogel CD-TFPP can effectively enter cells, and the cell uptake behavior is time-dependent.

[0058] The sonosensitizer nanogel provided by the present invention is subjected to an in vitro cytotoxicity experiment, and the experimental results show that the sonosensitizer nanogel CD-TFPP has the effect of inhibiting the growth and reproduction of cancer cells, and exhibits a good in vitro anti-tumor effect.

[0059] The sonosensitizer nanogel CD-TFPP provided by the present invention is used to improve the water solubility of the sonosensitizer and enhance the biocompatibility of the sonosensitizer on the one hand, and on the other hand, the large-sized cavity provides cavitation nuclei for the sonosensitizer to enhance the sonodynamic efficiency. At the same time, the simple synthesis method can conveniently adjust the ratio of the sonosensitizer and the cavitation nuclei and the size of the nanogel. In addition, the introduction of the main molecule can conveniently carry other therapeutic drugs that cooperate with sonodynamic therapy to realize a personalized treatment plan with easy-to-customize functions.

[0060] The following describes the preferred embodiments of the present invention with reference to the drawings in the specification to prove that the present invention can be implemented. The embodiments can fully introduce the present invention to those skilled in the art, making its technical content clearer and easier to understand. The present invention can be embodied in many different forms of embodiments, and the protection scope of the present invention is not limited to the embodiments mentioned in the text.

[0061] Example 1

[0062] Preparation method of CD-TFPP sonosensitizer:

[0063] This embodiment adopts a nucleophilic substitution synthesis method to prepare a customizable water-soluble sonosensitizer nanogel, comprising:

[0064] β-CD and 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin were reacted at a certain ratio (molar ratio, β-CD:TFPP=1:1.5, β-CD:TFPP=1:1, β-CD:TFPP=1:0.5), anhydrous potassium carbonate was used as a catalyst, N-methylpyrrolidone was used as a solvent, the reaction temperature was 85°C, and the reaction time was 40h. After the reaction, the product was dialyzed in ultrapure water using a dialysis bag with a molecular weight cutoff of 8000-14000 and freeze-dried to obtain a reddish brown product.

[0065] The reddish-brown product was redissolved in dimethyl sulfoxide to make it dissolve evenly, and the evenly mixed organic phase solution was slowly dripped into the aqueous phase which was stirred at high speed and kept at a constant temperature of 25°C using a syringe. The volume ratio of the organic phase to the aqueous phase was 1:4, and stirring was continued for one hour.

[0066] The stirred liquid was passed through a 220 nm aqueous membrane to obtain a clear CD-TFPP nanogel solution. According to different feed ratios (β-CD:TFPP=1:1.5, β-CD:TFPP=1:1, β-CD:TFPP=1:0.5), the obtained different nanoparticles were named CD-TFPP-1, CD-TFPP-2, and CD-TFPP-3.

[0067] Figure 1 The UV-visible absorption spectra of CD-TFPP aqueous solutions (CD-TFPP-1, CD-TFPP-2, CD-TFPP-3) with different molar ratios are shown, indicating that β-cyclodextrin successfully reacts with 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin. The instrument model used for the test is Evolution 220UV-Vis, and the spectrum scanning range is from 300nm to 700nm.

[0068] Example 2

[0069] Determination of fluorescence emission spectrum of CD-TFPP nanoparticles:

[0070] In this example, the fluorescence emission spectra of CD-TFPP nanoparticles of different sizes prepared in Example 1 were tested. A certain amount of CD-TFPP-1, CD-TFPP-2, and CD-TFPP-3 were prepared into deionized water solutions of the same concentration (prepared according to the operating specifications and instrument requirements), and according to their absorption curves in the ultraviolet spectrum, 415 nm was selected as the excitation wavelength, and then the excitation spectrum was measured using a fluorescence spectrophotometer. The results are shown in FIG. Figure 2 As shown, it can be seen that as the amount of modified cyclodextrin increases, its fluorescence emission is significantly enhanced, indicating that the steric hindrance of a large amount of cyclodextrin can avoid fluorescence quenching caused by the stacking of porphyrins and has high photostability.

[0071] Example 3

[0072] Determination of particle size distribution of CD-TFPP nanoparticles:

[0073] In this example, dynamic light scattering was used to measure the particle size distribution of CD-TFPP nanoparticles dispersed in an aqueous solution. The specific operation was as follows: the CD-TFPP nanoparticle aqueous solution prepared in Example 1 was added to a cuvette dedicated to measuring hydration kinetic particle size, and a dynamic light scattering instrument was used to detect its hydration particle size and dispersion index. The results are shown in FIG. Figure 3 As shown, the size distribution of CD-TFPP-1, CD-TFPP-2, and CD-TFPP-3 nanoparticles is 35nm, 75nm, and 110nm, respectively. Nanoparticles of different sizes can be easily assembled with other specific objects to form nanomedicines of different particle sizes, which is customizable.

[0074] Example 4

[0075] Determination of acoustic stability of CD-TFPP nanoparticles:

[0076] In this example, the nanoparticles CD-TFPP-1, CD-TFPP-2, and CD-TFPP-3 prepared in Example 1 were subjected to UV-visible light absorption spectrum measurement under continuous ultrasound. Figure 4 As shown in Figure 2, the UV absorption spectra of CD-TFPP-1, CD-TFPP-2, and CD-TFPP-3 nanoparticles remained almost unchanged within 30 min of continuous ultrasonic treatment (ultrasonic parameters were 1.0 MHz, 50% duty cycle, and 2.5 W·cm -2 ). Figure 4 The line graph of the acoustic stability of the nanoparticles shows that they can exist stably under ultrasonic treatment.

[0077] Example 5

[0078] Determination of Singlet Oxygen in Water and Nanoparticles in Vitro:

[0079] In this example, 1,3-diphenylisobenzofuran (DPBF) was used as a singlet oxygen scavenger to detect whether the nanoparticles could generate singlet oxygen after ultrasound. The ultrapure water and DPBF solution of the PBS group were added to a quartz test tube and ultrasound was applied for 30 seconds each time (ultrasound parameters were 1.0 MHz, 50% duty cycle, 2.5 W·cm -2 ), monitor the UV-visible absorption spectrum of DPBF at 425nm, such as Figure 5 As shown, the efficiency of singlet oxygen generation in a simple aqueous solution can be evaluated.

[0080] Similarly, taking the sample CD-TFPP-3 nanoparticles prepared in Example 1 as an example, the DPBF aqueous solution containing CD-TFPP-3 nanoparticles was treated with ultrasound for 30 s (ultrasound parameters were 1.0 MHz, 50% duty cycle, 2.5 W·cm -2 ), the absorbance of the DPBF aqueous solution at 425 nm was measured every 30 s to obtain a set of absorbance decreasing curves.

[0081] The concentration of the sample solution was 1.0×10 -5 M.

[0082] like Figure 6 , Figure 7 , Figure 8 and Fig. 9 As shown, the extent of DPBF degradation by CD-TFPP nanoparticles under ultrasound conditions is much greater than that of water in the control group, indicating that CD-TFPP nanoparticles have the effect of generating singlet oxygen by sonodynamics, which can effectively enhance the killing effect on tumors.

[0083] Example 6

[0084] Determination of CD-TFPP nanogel stability:

[0085] In this example, the CD-TFPP nanoparticles (prepared in Example 1) dispersed in the aqueous solution were left to stand for several days, during which the particle size of the CD-TFPP nanoparticles was measured using a dynamic light scattering instrument. Fig.10 As shown, it was shown that CD-TFPP nanoparticles could be stable for fifteen days.

[0086] Example 7

[0087] Uptake of CD-TFPP nanoparticles in cells:

[0088] In this example, three different CD-TFPP nanoparticles (prepared in Example 1) were subjected to cell uptake experiments. In this example, confocal laser scanning microscopy (CLSM) was used to study the cell uptake behavior of 4T1 tumor cells for each sample. 4 Cells / well) were inoculated into glass-bottomed culture dishes and cultured for 24 hours. Then the cells were cultured for 24 hours with fresh culture medium containing sonosensitizers with the same concentration of porphyrins (CD-TFPP-1, CD-TFPP-2, CD-TFPP-3, 20 μg / mL). The cells were carefully washed with PBS and then treated with 4% paraformaldehyde. After removing the paraformaldehyde, the cell nuclei were stained with Hoechst for 3 minutes and washed three times with PBS to terminate the staining. Finally, the fluorescence of porphyrins in the cells was observed by CLSM under 404 nm excitation and 660 nm emission. Fig.11 As shown, after 24 hours of cell incubation, red fluorescence intensity of porphyrin appeared in each group of cell samples, indicating that tumor cells can effectively take up nanoparticles of different sizes.

[0089] Example 8

[0090] In vitro cytotoxicity assay verification:

[0091] In this example, the biocompatibility of CD-TFPP prepared in Example 1 was studied. The toxicity of CD-TFPP to L929 cells was studied using the MTT method. L929 cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) containing 1% penicillin / streptomycin and 10% fetal bovine serum. The temperature was constant (37°C), and the culture environment was humid and contained 5% carbon dioxide. The MTT method studied the cytotoxicity of the two samples to L929 cells. In a 96-well plate, 200 μL of DMEM culture medium was added to each well, and the cells were seeded in a 96-well plate at a density of 5000 per well. After incubation at 37°C for 24 hours, the culture medium was discarded and washed with PBS. Then, different concentrations of samples (dissolved in fresh DMEM culture medium) were added to each well, and all samples were incubated with cells for 24 hours. The next day, the culture medium containing the sample was washed away with PBS, and then washed with 5 mg mL -1 The MTT-containing RPMI DMEM medium was further incubated for 4 hours. After discarding the medium, 150 μL DMSO was added to each well to extract the formazan product. Finally, the absorbance (OD) at 492 nm was measured using a spectrophotometric microplate reader (Thermo Multiskan MK3 spectrometer).

[0092] The calculation method of cell viability is as follows (1-1):

[0093] Cell viability (%) = (OD test ) / (OD control )×100(1-1);

[0094] OD test is the absorbance of the sample group, OD control is the absorbance of the untreated cell group.

[0095] like Fig.12 As shown in the figure, the cell activity diagram after CD-TFPP nanoparticles were co-incubated with L929 cells showed that CD-TFPP nanoparticles had almost no toxicity in a normal cell environment, especially when the porphyrin concentration in the nanogel was as high as 300 μg / mL, more than 80% of L929 cells still survived. This result shows that CD-TFPP nanoparticles have good biocompatibility.

[0096] Similarly, various cancer cell lines were used as research objects, and the cytotoxicity was tested by MTT method, and the absorbance was measured by spectrophotometric microplate reader. Various cancer cell lines were used as research objects, and the cytotoxicity of CD-TFPP nanoparticles under conditions without and with ultrasonic treatment was measured by MTT method, respectively. Fig.13 and Fig.14 The data of dark cytotoxicity and acoustic toxicity of the cells are shown in Figure 2. The experiment shows that: in the absence of ultrasound ( Fig.13 ), CD-TFPP nanoparticles have little toxicity to cells even at higher concentrations. Fig.14 , the ultrasonic parameters were 1.0 MHz, 50% duty cycle, 1.0 W·cm -2 ), with the increase of porphyrin concentration, the cell activity gradually decreased, that is, the SDT effect gradually increased.

[0097] Example 9

[0098] Intracellular SDT Effects:

[0099] In order to further verify the effect of sonosensitizer CD-TFPP nanoparticles on cells, this example takes CD-TFPP-3 prepared in Example 1 as an example and conducts AM / PI staining experiments. Fig.15 The results showed that the PBS group and CD-TFPP-3 group without ultrasound treatment showed obvious green fluorescence, indicating that the sonosensitizer CD-TFPP-3 was almost non-toxic to cells. In the PBS ultrasound group, the mechanical force of ultrasound also caused cell damage. In the CD-TFPP-3 ultrasound group, a clear red color was shown, indicating that almost all cells died. The sonosensitizer CD-TFPP had a good effect in sonodynamic therapy.

[0100] In summary, the present invention provides a water-soluble sonosensitizer nanogel with easily customizable functions and a preparation method thereof. The sonosensitizer nanogel (CD-TFPP) is prepared by nucleophilic substitution of β-cyclodextrin and 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin (TFPP). The sonosensitizer nanogel introduces a macrocyclic β-cyclodextrin (β-CD) with an inherent cavity, which not only improves the water solubility and stability of porphyrin, but also provides a cavitation nucleus for the porphyrin sonosensitizer, thereby obtaining a higher singlet oxygen yield and sonodynamic efficiency. In addition, the nanogel containing the main molecule can be used as a drug-carrying platform, carrying different drug-acting guest molecules as needed, providing new opportunities for personalized tumor treatment.

[0101] The above description of the embodiments is to facilitate the understanding and use of the invention by those skilled in the art. It is obvious that those skilled in the art can easily make various modifications to these embodiments and apply the general principles described herein to other embodiments without creative work. Therefore, the present invention is not limited to the above embodiments, and improvements and modifications made by those skilled in the art based on the disclosure of the present invention without departing from the scope of the present invention should be within the scope of protection of the present invention.

Claims

1. A water-soluble sonosensitizer nanogel, characterized in that: The nanogel has the following structural formula I: Wherein, R is H or 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin and at least one R is 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin, and n is a positive integer.

2. A water-soluble sonosensitizer nanogel according to claim 1, characterized in that: The size of the nano gel is 35-110 nm.

3. A method for preparing a water-soluble sonosensitizer nanogel according to claim 1 or 2, characterized in that: The nano gel is prepared from a cyclic compound beta-cyclodextrin with a large cavity and a sonosensitizer 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin with a sonosensitizing effect through a nucleophilic substitution reaction.

4. The method for preparing a water-soluble sonosensitizer nanogel according to claim 3, characterized in that: The steps include: S1: in a first solvent, β-cyclodextrin and 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin react under the condition of a catalyst, and then dialyze and freeze-dry to obtain a reaction product; S2: dissolving the reaction product solvent obtained in step S1 in a second solvent to form an organic phase, and dripping the organic phase into the water phase stirred at a constant temperature until the mixed solution is stable; S3: filtering the mixed solution obtained in step S3 through an aqueous phase membrane to obtain the nanogel.

5. The method for preparing a water-soluble sonosensitizer nanogel according to claim 4, characterized in that: In step S1, the first solvent includes N-methylpyrrolidone, the catalyst includes anhydrous potassium carbonate, and the molar ratio of β-cyclodextrin to 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin is 1:0.5-1.

5.

6. The method for preparing a water-soluble sonosensitizer nanogel according to claim 4, characterized in that: In step S1, the reaction temperature is 80-90°C and the reaction time is 35-50 hours; the dialysis is performed in ultrapure water using a dialysis bag with a molecular weight cutoff of 8000-14000 and the dialysis time is 48-60 hours.

7. The method for preparing a water-soluble sonosensitizer nanogel according to claim 4, characterized in that: In step S2, the second solvent includes dimethyl sulfoxide; the volume ratio of the organic phase to the aqueous phase is 1:3-5.

8. The method for preparing a water-soluble sonosensitizer nanogel according to claim 4, characterized in that: In step S2, the temperature of the water phase is 20-30°C.

9. The method for preparing a water-soluble sonosensitizer nanogel according to claim 4, characterized in that: In step S3, the aqueous phase membrane is an aqueous phase membrane of 200 to 300 nm.

10. Use of the water-soluble sonosensitizer nanogel according to claim 1 or 2 in the preparation of sonosensitizers.

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