Preparation method and application of a nano diagnosis and treatment platform

Abstract

The present invention discloses a preparation method and application of a nano-diagnosis and treatment platform, which relates to the technical field of bioimaging and treatment. The nano-diagnosis and treatment platform of the present invention uses a fluorinated amphiphilic organic polymer as a carrier, which is tightly combined with a fluorinated organic semiconductor photosensitizer through a self-assembly process to obtain nanoparticles with good hydrophobicity and biocompatibility. These nanoparticles, by virtue of their unique intermolecular fluorine-fluorine interaction and the ability of fluorine atoms to capture oxygen, achieve an improvement in fluorescence quantum yield and significantly enhance the 1 generation ability of O2. The nano-diagnosis and treatment platform of the present invention not only exhibits excellent NIR-II FLI performance, but also has efficient PDT and photothermal conversion capabilities, providing strong technical support for the realization of precise and efficient tumor photodynamic / photothermal combined therapy.

Description

Technical Field

[0001] The present invention relates to the technical field of bioimaging and therapy, and particularly to a preparation method and application of a nanodiagnosis and treatment platform. Background Art

[0002] Compared with traditional surgery, radiotherapy, and chemotherapy, optical diagnosis and treatment technologies can detect cancer cells at an earlier stage. Through optical imaging technologies such as fluorescence imaging and photoacoustic imaging, three-dimensional visualization of tumor tissues can be achieved, providing detailed lesion information for doctors to formulate more personalized treatment plans. In addition, advanced optical treatment methods such as PDT (photodynamic therapy) and PTT (photothermal therapy) can accurately act on cancer cells while reducing damage to surrounding normal tissues, thereby reducing the toxic and side effects during treatment and significantly improving the quality of life of patients. These technologies are not only applicable to the treatment of superficial tumors but also continuously break through limitations and expand into the field of deep tumor treatment by using various innovative means, such as expanding the absorption wavelength range of photosensitizers, enhancing the penetration depth of light in tissues, and promoting the targeted enrichment of photosensitizers at the tumor site. With the continuous progress and deep integration of nanoscience and biotechnology, a series of more precise, efficient, and low-side-effect cancer treatment plans have been developed successively.

[0003] Near-infrared II (NIR-II, 1000 - 1700 nm) fluorescence imaging technology (NIR-II FLI), with its superior penetration depth, high-resolution imaging ability, and the advantage of real-time dynamic monitoring of the internal structure and function changes of organisms, is gradually becoming a cutting-edge tool for organism visualization. In promoting the clinical translation process of NIR-II FLI, it is crucial to develop photosensitizers that integrate high brightness (high fluorescence quantum yield and molar extinction coefficient ε), long wavelength, high photostability, and biocompatibility.

[0004] As an innovative cancer treatment method, the core principle of PDT is to use photosensitizers to convert light energy into reactive oxygen species (ROS) to induce apoptosis and tissue damage. Although type II PDT dominates due to its efficient oxygen-dependent 1 O2 generation mechanism, its efficacy is limited by the hypoxic environment inside tumors. To break through this bottleneck, researchers are actively exploring various strategies to enhance the treatment effect of PDT under hypoxic conditions. For example, by hyperbaric oxygen therapy or developing nano-carriers for oxygen delivery to increase the oxygen content in tumors to increase 1Generation of O2; develop type I photosensitizers that do not rely on oxygen or dual-mechanism photosensitizers that combine type I and type II mechanisms to effectively kill tumor cells in hypoxic environments; at the same time, continuously optimize the performance of photosensitizers, such as increasing the fluorescence quantum yield and molar extinction coefficient, improving photostability, and enhancing biocompatibility, to enhance the overall efficacy of PDT. In addition, PDT is also combined with other treatment methods such as PTT, chemotherapy, radiotherapy, and immunotherapy to further improve the treatment effect and reduce side effects using synergistic effects.

[0005] Fluorination modification exhibits significant advantages in the biomedical field, especially in reducing physiological toxicity and enhancing treatment effects. Its unique advantages stem from the chemical properties of fluorine atoms, namely their large electronegativity and small atomic radius, which endow fluorinated compounds with higher stability and lower reactivity in living organisms, thereby effectively reducing the toxic and side effects on normal cells. At the same time, fluorination can stabilize the electronic structure of fluorescent materials, reduce non-radiative transitions, and thus increase the fluorescence quantum yield and optical properties. In addition, fluorinated polymers have excellent oxygen capture ability. Therefore, adopting the fluorination strategy is expected to improve the hypoxic environment inside tumors, provide more oxygen sources for PDT, and thus significantly improve the treatment effect. Amphiphilic polymers have been widely used in drug delivery systems due to their unique structure (containing both hydrophilic and hydrophobic parts). As an effective terminal modification method, fluorination modification can not only enhance the stability and biocompatibility of polymer carriers but also endow them with new functional characteristics. These improvements make fluorinated amphiphilic polymer carriers show broader application potential and prospects in advanced photodiagnosis and therapy fields. Summary of the Invention

[0006] The purpose of the present invention is to provide a preparation method and application of a nano-diagnosis and treatment platform to solve the problems existing in the above-mentioned prior art.

[0007] To achieve the above purpose, the present invention provides the following solutions:

[0008] One of the purposes of the present invention: Provide an amphiphilic fluorinated organic polymer with a general formula structure as shown in formula (I):

[0009]

[0010] In formula (I), the value range of z is 11 - 230; m, x, and y are polymerization degrees, the value range of m is 5 - 200, the value range of x is 5 - 200, and the value range of y is 5 - 200.

[0011] More preferably, the value of z is 11, 23, 45, 113, or 226.

[0012] Another purpose of the present invention: Provide a preparation method of the above amphiphilic fluorinated organic polymer, including the following steps:

[0013] A. Under an oxygen - free and water - free environment, a commercial compound A with the structural formula , a commercial compound B with the structural formula

[0014] , and the catalyst 4 - dimethylaminopyridine (DMAP) are mixed. Tetrahydrofuran (THF) is used as the reaction solvent, and the mixture is refluxed under heating and stirring. After the reaction is completed, the reaction solution is separated and purified to obtain the amphiphilic polymer PSMA - PEG with the structural formula ;

[0015] B. The amphiphilic polymer PSMA - PEG and N,N` - succinimidyl carbonate (DSC) are added to anhydrous dichloromethane (DCM), and then N,N - diisopropylethylamine (DIPEA) is added, and the mixture is stirred at room temperature for 6 h;

[0016] C. An excess of amino - trifluoroaniline (NH2 - ph - 3F) is added to the reaction system of step B, and the mixture is continuously stirred at room temperature for 48 h. After the reaction is completed, the solvent is removed, the solid product is dissolved in deionized water, dialyzed, and freeze - dried to obtain the amphiphilic fluorinated organic polymer PSMA - 3F.

[0017] Among them, the value range of z is 11 - 230; m, x, and y are the degree of polymerization, the value range of m is 5 - 200, the value range of x is 5 - 200, and the value range of y is 5 - 200.

[0018] More specifically, the polyethylene glycol grafted on the main chain is amino - polyethylene glycol - methoxy with different molecular weights such as 500, 1000, 2000, 5000, 10000, etc.

[0019] Furthermore, in step A, the temperature of heating and stirring is 70 °C, and the time is 24 h; the molar ratio of the amphiphilic polymer PSMA - PEG to N,N` - succinimidyl carbonate is 1:2 - 2:3; the molar ratio of the amphiphilic polymer PSMA - PEG to amino - trifluoroaniline is 1:1.2 - 1:2.

[0020] The third object of the present invention: To provide a fluorinated organic photosensitizer 4F having a general formula structure as shown in formula (II):

[0021]

[0022] In formula (II), R and R1 are each independently selected from branched or straight - chain C4 - C24.

[0023] The fourth object of the present invention: To provide a preparation method of the above - mentioned fluorinated organic photosensitizer,

[0024] The one with the structural formula Compound 1 and the structural formula is Compound 2 was dissolved in chloroform under an argon atmosphere, then pyridine was added, and the reaction mixture was heated under reflux at 55 °C for 16 h. After the reaction was completed, when the reaction mixture was cooled to room temperature, it was poured into methanol to precipitate the crude product, which was collected by filtration and purified to obtain the fluorinated organic photosensitizer;

[0025] Wherein, R and R1 are each independently selected from branched or straight-chain C4-C24.

[0026] More specifically, it includes the following steps:

[0027] The structural formula is Compound 1 (0.1 mmol) and the structural formula is Compound 2 (0.5 mmol) were dissolved in chloroform (30 mL) under an argon atmosphere. Then an appropriate amount of pyridine (0.6 mL) was added, and the reaction mixture was heated under reflux at 55 °C for 16 h. After the reaction was completed, when the reaction mixture was cooled to room temperature, it was poured into methanol to precipitate the crude product, which was collected by filtration. The crude product was further purified by silica gel column chromatography using petroleum ether:dichloromethane (1:1) as the eluent to obtain a black solid 4F with the structural formula

[0028] Wherein, R and R1 are each independently selected from branched or straight-chain C4-C24.

[0029] The fifth object of the present invention: To provide the application of the above amphiphilic fluorinated organic polymer as a carrier of a nanodiagnosis and treatment platform.

[0030] The sixth object of the present invention: To provide the application of the above fluorinated organic photosensitizer as a photosensitizer of a nanodiagnosis and treatment platform.

[0031] The seventh object of the present invention: To provide a nanodiagnosis and treatment platform prepared by the above amphiphilic fluorinated organic polymer and fluorinated organic photosensitizer by a nano-coprecipitation method; the coprecipitation method includes the following steps: mixing the above amphiphilic fluorinated organic polymer and fluorinated organic photosensitizer in an aqueous tetrahydrofuran solution, ultrasonicating, and removing the solvent under a protective atmosphere to obtain water-soluble nanoparticles, which are the nanodiagnosis and treatment platform (FF NPs).

[0032] More preferably, the mass ratio of the amphiphilic fluorinated organic polymer to the fluorinated organic photosensitizer is 5:1 to 8:1.

[0033] The eighth object of the present invention: To provide the application of the above nanodiagnosis and treatment platform in the preparation of tumor photothermal and / or photodynamic therapy reagents.

[0034] Ninth objective of the present invention: To provide the application of the above-mentioned nano-diagnostic and treatment platform as a photosensitizer for tumor photothermal and / or photodynamic therapy, and as a contrast agent for near-infrared second-window fluorescence imaging (NIR-II FLI) of tumors.

[0035] The present invention successfully constructs a nano-diagnostic and treatment platform by co-assembling the prepared fluorine-rich amphiphilic organic polymer as a carrier with a fluorinated organic semiconductor small molecule having excellent light absorption ability. This nano-diagnostic and treatment platform utilizes the fluorine-fluorine interaction between molecules to greatly improve the fluorescence quantum yield (QY), and due to its abundant fluorine atoms, it has stronger oxygen capture ability, effectively enhancing 1 the generation of O2.

[0036] The nano-diagnostic and treatment platform of the present invention is prepared by encapsulating a fluorinated photosensitizer in a fluorine-containing amphiphilic organic polymer modified with NH2-ph-3F (formed by copolymerization of PSMA and NH2-PEG). This nano-diagnostic and treatment platform has enhanced NIR-II FLI and PDT effects due to fluorination. The preparation process of this nano-diagnostic and treatment platform first modifies amino-polyethylene glycol (NH2-PEG) on isopropylbenzene-capped poly(styrene-co-maleic anhydride) (PSMA) through an amidation reaction to obtain an amphiphilic polymer carrier. Subsequently, through a further amidation reaction, the fluorine source molecule amino-trifluoroaniline is introduced into the PSMA-PEG polymer backbone to construct a fluorine-containing amphiphilic polymer nanocarrier PSMA-3F. Then, the above-mentioned fluorinated organic photosensitizer (abbreviated as 4F) is combined with the polymer nanocarrier PSMA-3F by a nano-precipitation method in a mixed solvent of water and tetrahydrofuran to prepare water-soluble nanoparticles FF NPs. These nanoparticles exhibit enhanced NIR-II FLI performance in water, and under 808 nm laser irradiation, they exhibit enhanced singlet oxygen ( 1 O2) generation ability and good photothermal conversion efficiency. Therefore, this nano-diagnostic and treatment platform is expected to achieve the synergistic treatment of tumor PDT and photothermal therapy (PTT) under the guidance of NIR-II FLI.

[0037] The present invention provides the application of the above-mentioned nano-diagnostic and treatment platform as a contrast agent for tumor NIR-II FLI. Specifically, the near-infrared second-window fluorescence imaging method: Prepare an aqueous solution of FF NPs with a certain concentration, place it in a small centrifuge tube, and under an NIR-II fluorescence imaging instrument, excite it with an 808 nm laser to obtain an NIR-II FLI image.

[0038] The present invention provides the application of the above-mentioned nano-diagnostic and treatment platform as a photosensitizer for tumor photothermal and / or photodynamic therapy. Specifically, in terms of the application of photodynamic therapy reagents, it includes the following test steps:

[0039] A certain concentration of FF NPs aqueous solution was added to the cuvette, and then 1 the O2 indicator (SOSG, 1 - 10 μM) was added dropwise. A laser with a set light power density (808 nm, 0.1, 0.25, 0.5, 0.75, 1 W cm -2 ) was used to intermittently irradiate the sample, and the fluorescence intensity at 521 nm in the fluorescence spectrum under illumination at different times was recorded by a spectrometer, and a curve of the change with illumination time was plotted. At the same time, a blank group (only adding the same concentration of SOSG) and a control group (using PF NPs aqueous nano - solution plus the same concentration of SOSG) were set up, and the experimental operations were carried out according to the same steps and the data was recorded.

[0040] Specifically, in the application of the photothermal therapy reagent, the following test steps are included:

[0041] Small centrifuge tubes containing deionized water (blank group) and small centrifuge tubes containing the same concentration of FF NPs or PF NPs aqueous solution were respectively irradiated with a laser with a certain light power density, and then they were cooled to room temperature. At the same time, an infrared thermosensitive thermal imager was used to record the trend of the solution temperature change with time in these two centrifuge tubes.

[0042] Among them, the PF NPs aqueous solution was prepared by directly encapsulating the fluorinated organic photosensitizer 4F with the fluorine - free amphiphilic organic polymer PSMA - PEG.

[0043] The present invention discloses the following technical effects:

[0044] The nano - diagnosis and treatment platform of the present invention is centered on using the fluorinated amphiphilic organic polymer as a carrier, which is tightly combined with the fluorinated organic semiconductor photosensitizer through a self - assembly process to obtain nanoparticles with good hydrophobicity and biocompatibility. These nanoparticles, relying on their unique intermolecular fluorine - fluorine interaction and the oxygen - capturing ability of fluorine atoms, have achieved an improvement in the fluorescence quantum yield and significantly enhanced the 1 ability to generate O2.

[0045] The nano - diagnosis and treatment platform of the present invention not only exhibits excellent NIR - II FLI performance but also has high - efficiency PDT and photothermal conversion capabilities, providing strong technical support for realizing precise and efficient tumor photodynamic / photothermal combined therapy.

[0046] The fluorination - enhanced nano - diagnosis and treatment platform of the present invention, with excellent imaging quality and significant treatment effects, can achieve the synergistic therapy of PDT and PTT under the precise guidance of NIR - II FLI, opening up a new way for the treatment of diseases such as inflammation and tumors, and bringing a safer and more effective treatment plan for patients. Brief Description of the Drawings

[0047] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can be obtained based on these drawings.

[0048] Figure 1 Schematic diagram of the preparation process of the nano-diagnosis and treatment platform FF NPs of the present invention.

[0049] Figure 2 1H NMR spectrum of the fluorinated organic photosensitizer 4F prepared in Example 1 of the present invention 1 H NMR spectrum.

[0050] Figure 3 MALDI-TOF mass spectrum of the fluorinated organic photosensitizer 4F prepared in Example 1 of the present invention.

[0051] Figure 4 1H NMR spectra of the fluorinated amphiphilic organic polymer carrier PSMA-3F and its non-fluorinated control group PSMA-PEG prepared in Example 1 of the present invention 1 H NMR spectrum.

[0052] Figure 5 Gel permeation chromatography (GPC) diagrams of the fluorinated amphiphilic organic polymer carrier PSMA-3F and its non-fluorinated control group PSMA-PEG prepared in Example 1 of the present invention.

[0053] Figure 6 UV absorption and NIR-II fluorescence emission spectra of FF NPs and PF NPs.

[0054] Figure 7 NIR-II FLI diagrams of FF NPs and PF NPs.

[0055] Figure 8 When 10 μM aqueous solutions of FF NPs and PF NPs are irradiated with a laser of the same optical power density (808 nm, 0.5 W / cm 2 ), the respective 1 generation of O2.

[0056] Figure 9 When 12 μM aqueous solutions of FF NPs and PF NPs are irradiated with a laser of the same optical power density (808 nm, 0.5 W / cm 2 ), the photothermal heating and cooling conditions.

[0057] Figure 10Confocal fluorescence imaging of the cellular uptake of FF NPs or PF NPs by 4T1 cells (a) and the quantitative analysis results of their fluorescence intensities (b).

[0058] Figure 11 Photodynamic / photothermal synergistic killing analysis test of FF NPs or PF NPs on 4T1 cells.

[0059] Figure 12 In vivo NIR-II tumor fluorescence imaging of a live mouse after tail vein injection of FF NPs (a) and the curve of the fluorescence intensity at the tumor site over time (b). Detailed implementation manners

[0060] The various exemplary implementation manners of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, characteristics, and implementation schemes of the present invention.

[0061] It should be understood that the terms described in the present invention are only for describing specific implementation manners and are not used to limit the present invention. Additionally, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded from the range.

[0062] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention pertains. Although the present invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein can also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0063] Without departing from the scope or spirit of the present invention, various improvements and changes can be made to the specific implementation manners of the present invention specification, which are obvious to those skilled in the art. Other implementation manners obtained from the present invention specification are also obvious to those skilled in the art. The present invention specification and examples are only exemplary.

[0064] Regarding the terms "comprising", "including", "having", "containing", etc. used herein, they are all open-ended terms, meaning including but not limited to.

[0065] Figure 1 Schematic diagram of the preparation process of the FF NPs of the nano-diagnosis and treatment platform of the present invention.

[0066] Specific descriptions will be given below in conjunction with embodiments.

[0067] Example 1

[0068] (1) Synthesis of fluorinated organic photosensitizer 4F:

[0069]

[0070] Compound 1 (0.1 mmol) with the structural formula and compound 2 (0.5 mmol) with the structural formula were dissolved in chloroform (30 mL) under an argon atmosphere. Then, an appropriate amount of pyridine (0.6 mL) was added, and the reaction mixture was heated under reflux at 55 °C for 16 h. After the reaction was completed, the reaction mixture was cooled to room temperature and poured into methanol to precipitate the crude product, which was collected by filtration. The crude product was further purified by silica gel column chromatography using petroleum ether:dichloromethane (1:1) as the eluent to obtain the black solid 4F with the structural formula .

[0071] Figure 2 The 1 1H NMR spectrum of the fluorinated organic photosensitizer 4F prepared in Example 1 Figure 3 is shown in Figure 28, and

[0072] (2) Synthesis of PSMA-PEG:

[0073]

[0074] Poly(styrene-co-maleic anhydride) (end-capped with cumene, 0.1 mmol, Mn ~ 1900), amino-polyethylene glycol (Mn ~ 5000, 0.3 mmol), and the catalyst DMAP were added to a reaction flask. Subsequently, sufficient THF was added to the reaction flask as the reaction solvent. The mixture was stirred and heated to 70 °C, and condensed reflux was carried out to prevent solvent evaporation. The reaction was carried out under these conditions for 24 hours. After the reaction was completed, the reaction mixture was cooled to room temperature and precipitated with ether. Subsequently, the precipitate was redissolved in deionized water and freeze-dried to obtain the amphiphilic polymer PSMA-PEG (Mn ~ 6974 kDa).

[0075] (3) Synthesis of PSMA-3F:

[0076]

[0077] The product PSMA-PEG (0.1 mmol) and DSC (0.2 mmol) were added into a reaction flask, and sufficient anhydrous DCM was added as the reaction solvent. Then, a small amount of DIPEA was slowly added dropwise as a catalyst. At room temperature, the reaction mixture was continuously stirred for 6 hours. Subsequently, an excess of NH2-ph-3F (0.2 mmol) was added to the above-mentioned reaction flask that had been stirred for 6 hours, and stirring was continued at room temperature for 48 hours to introduce a fluorine source into the polymer backbone.

[0078] After the reaction was completed, the crude product was dried by rotary evaporation. Subsequently, the solid substance was dissolved in deionized water and then dialyzed in deionized water using a dialysis bag (MW 7000 Da) for 3 days. Finally, it was freeze-dried to obtain the amphiphilic fluorinated organic polymer carrier PSMA-3F (Mn ~ 9714 kDa).

[0079] Figure 4 For the fluorinated amphiphilic organic polymer carrier PSMA-3F prepared in Example 1 and its PSMA-PEG (non-fluorinated control group) 1 1H NMR spectra, Figure 5 are their gel permeation chromatography (GPC) diagrams.

[0080] (3) Preparation of the nanotheranostic platform: By the nano-coprecipitation method, the PSMA-3F prepared above and 4F (mass ratio 5:1) were placed in a binary solution of THF and deionized water (volume ratio 1:9), and nanoparticles FFNPs were obtained under ultrasonic action. Subsequently, THF was removed by evaporation under nitrogen protection.

[0081] Comparative Example 1

[0082] The difference from Example 1 was only that the fluorine-free amphiphilic organic polymer PSMA-PEG was directly used to encapsulate 4F to obtain PF NPs:

[0083] By the nano-coprecipitation method, the PSMA-PEG prepared in Example 1 and 4F (mass ratio 5:1) were placed in a binary solution of THF and deionized water (volume ratio 1:9), and nanoparticles PF NPs were obtained under ultrasonic action. Subsequently, THF was removed by evaporation under nitrogen protection.

[0084] Performance testing:

[0085] (1) UV absorption and fluorescence emission tests of FF NPs and PF NPs were carried out as follows:

[0086] Take 3 mL of aqueous solutions of FF NPs and PF NPs with the same concentration and place them in two cuvettes respectively. Subsequently, put them into an ultraviolet spectrophotometer respectively to collect the images of the ultraviolet absorption spectra. Next, in the same way, place these two samples in a fluorescence spectrophotometer respectively to collect their respective emission spectra. The results are as Figure 6 shown.

[0087] From Figure 6 it can be observed that the maximum absorption wavelengths of FF NPs and PF NPs are basically the same, and both show strong absorption ability in the near-infrared region, which is attributed to the unique properties of the photosensitizer 4F. In addition, the emission wavelengths of both show typical NIR-II fluorescence emission, and its emission tail band extends to 1300 nm in aqueous solution, which can effectively avoid the fluorescence interference of in vivo imaging and is beneficial to high-contrast NIR-II FLI of deep tissues. It should be noted that at the same concentration, FF NPs show significantly higher fluorescence intensity than PF NPs, which clearly indicates the significant enhancement effect of fluorination on fluorescence performance.

[0088] (2) NIR-II FLI tests of FF NPs and PF NPs are carried out as follows:

[0089] Take 300 μL of aqueous solutions of FF NPs and PF NPs with the same concentration and place them in two 0.5 mL centrifuge tubes respectively. Subsequently, place them in a NIR-II fluorescence imager respectively, irradiate them with an 808 nm laser, and the power density is 0.5 W / cm 2 , with a 1000 nm long-pass filter, to collect fluorescence images. The results are as Figure 7 shown.

[0090] From Figure 7 it can be observed that there are obvious intensity differences in the NIR-II fluorescence imaging maps of FF NPs and PF NPs. The results are similar to the fluorescence spectra, and FF NPs show brighter fluorescence emission, which may be attributed to their own fluorination.

[0091] (3) The 1 O2 generation ability tests of FF NPs and PF NPs are carried out as follows:

[0092] The SOSG (Singlet Oxygen Sensor Green) singlet oxygen probe is used to detect 1 the generation of O2. Mix 10 μL of SOSG (working concentration: 5 μM) with the aqueous solution of FF NPs or PF NPs (working concentration: 10 μM). Use 10 μL of SOSG and 3 mL of deionized water as a control group. Then, expose the mixture to an 808 nm laser (0.5 W / cm2 ) Every 30 s of illumination, the fluorescence intensity at approximately 521 nm was recorded in a fluorescence spectrometer (excitation wavelength: 488 nm). The results are as Figure 8 shown.

[0093] Figure 8 The variation of the fluorescence intensity at 521 nm with the illumination time after the mixing of FF NPs or PF NPs with 1 O2 in the aqueous phase. As Figure 8 can be seen, when only SOSG is present, the fluorescence signal intensity of the solution shows almost no significant change after illumination. In contrast, when the FF NPs or PF NPs solution coexists with SOSG, the fluorescence signal intensity of the solution gradually increases, and this enhancement effect is more significant in the presence of FF NPs. This indicates that both FF NPs and PF NPs have good 1 O2 generation ability, and FF NPs exhibit more excellent performance in this regard, probably because the fluorinated FF NPs enhance their oxygen capture ability.

[0094] (4) The test of the photothermal conversion efficiency of FF NPs and PF NPs was carried out as follows:

[0095] Calculate the photothermal conversion efficiency of FF NPs and PF NPs. 300 μL of aqueous solutions of FF NPs and PF NPs with the same concentration were irradiated with an 808 nm laser (0.5 W / cm 2 ) for 5 min, and the temperature change during illumination was monitored. Subsequently, the laser was turned off, and the solution was cooled to room temperature, and the temperature change during the cooling process was continuously monitored. During this process, the temperature change of the solution was recorded every 30 s. Additionally, 300 μL of deionized water was set as a blank control group. The results are as Figure 9 shown.

[0096] According to Figure 9 the obtained data, the photothermal conversion efficiencies of FF NPs and PF NPs were calculated to be PCE FF NPs = 52.8% and PCE PF NPs = 51.6%, respectively. This indicates that FF NPs and PF NPs have excellent photothermal properties and show potential for in vivo tumor therapy.

[0097] (5) The cell uptake test of FF NPs or PF NPs was carried out as follows:

[0098] Mouse breast cancer cells (4T1) were seeded in confocal microscope dishes and incubated for at least 24 h. Then, FF NPs or PF NPs (20 μM) were added and incubated for 9 h. Meanwhile, a blank group without any materials was set as the Control group. After incubation, the cells were washed 3 times with PBS. Then, 100 μL of 1× Hoechst33342 nuclear stain was added to each well and incubated for 30 min at room temperature in the dark. Subsequently, the stain was removed and the cells were washed 3 times with PBS. Finally, the cells were observed under a confocal microscope (Zeiss) in sequential scanning mode. FF NPs or PF NPs (λex = 565 nm, λem = 780 nm), Hoechst33342 (λex = 346 nm, λem = 461 nm). The results are as Figure 10 shown.

[0099] Figure 10 Analysis of the uptake of FF NPs or PF NPs by 4T1 cells. The results of confocal imaging confirmed that FF NPs or PF NPs could be well taken up by the cells, and stronger fluorescence of FF NPs was observed in 4T1 cells after 9 h, which might be related to the longer retention time of FF NPs in the cells.

[0100] (6) Test of the photodynamic / photothermal synergistic killing ability of FF NPs or PF NPs on 4T1 cells. The specific steps are as follows:

[0101] A CCK-8 cell viability detection kit was used. 4T1 cells were cultured in DMEM complete medium (containing 10% FBS, 1% penicillin / streptomycin) in a humidified incubator (37 °C, 5% CO2, 20% O2). Then, cells in the logarithmic growth phase were harvested and seeded in 96-well plates at a density of 1×10 4 cells and incubated for 24 h. Subsequently, the medium was replaced with fresh complete medium containing different concentrations of FF NPs or PF NPs. After further incubation for 6 h, the cells were exposed to 808 nm laser irradiation for 5 min (0.5 W / cm 2 ), denoted as FF NPs+L or PF NPs+L, respectively. Meanwhile, cells incubated with FF NPs or PF NPs without laser irradiation were set as the dark control group. After further incubation for 18 h, the medium was removed and the cells were washed 3 times with PBS. Then, the cells were incubated with fresh serum-free basal medium containing 10% CCK-8 in the dark for 30 min. Finally, the absorbance of the product was measured at a wavelength of 450 nm using a microplate reader. The results were expressed as the percentage of cell survival relative to the control group of cells without any treatment. The results are as Figure 11 shown.

[0102] Figure 11Analysis and test of the photodynamic / photothermal synergistic killing ability of FF NPs or PF NPs against 4T1 cells. It can be seen that as the concentration of the nanomaterials increases, the cell viability in the non-irradiated group does not decrease. On the contrary, whether incubated with FF NPs or PF NPs, the viability of 4T1 cells decreases with the increase of the material concentration under light irradiation. When the concentration of both is 15 μM, the cell killing effect is obvious, and FF NPs show stronger cell killing ability. According to the aforementioned in vitro performance tests, it can be analyzed that under the condition of basically the same photothermal conversion efficiency, that is, almost the same photothermal effect, the stronger 1 O2 generation ability of FF NPs endows it with more excellent 4T1 cell killing ability. The results at the cellular level further verify that fluorinated FF NPs have excellent synergistic phototherapy effects and show more excellent tumor killing potential.

[0103] (7) NIR-II fluorescence imaging test of the tumor site in tumor-bearing mice:

[0104] To study the feasibility of using FF NPs for in vivo fluorescence imaging-guided tumor therapy, a solid tumor model was established by subcutaneous injection of 4T1 cells. After intravenous injection of the aqueous solution of FF NPs, fluorescence imaging maps of the tumor sites of the mice were collected at different time points (0 h, 1 h, 3 h, 6 h, 12 h, 24 h, 48 h) after injection. The results are as Figure 12 shown.

[0105] Due to the enhanced permeability and retention (EPR) effect of tumors, Figure 12 It can be seen that after intravenous injection of FF NPs, the tumor sites of the mice quickly showed bright fluorescence, and the fluorescence signal intensity reached the peak within 3 hours. Subsequently, although it decreased slightly, it still maintained a relatively obvious fluorescence brightness for a long time. This result indicates that FF NPs can quickly penetrate the tumor vascular wall and accumulate in the tumor tissue during blood circulation, and show excellent tumor retention performance. This indicates that these nanoparticles have great potential as fluorescence imaging probes to guide deep tumor photodynamic / photothermal therapy.

[0106] In summary, the synthesis steps of the cancer nanodiagnostic platform provided by the present invention for enhancing NIR-II FLI and PDT efficiency are simple, the separation and purification operations are simple, with obvious fluorescence enhancement and improved 1 O2 generation ability; high-efficiency photothermal conversion performance, strong killing ability against breast cancer cells, can be used for in vivo vascular imaging, and has application potential in constructing anti-cancer drugs with efficient NIR-II FLI-guided photodynamic / photothermal synergistic therapy effects.

[0107] The embodiments described above are only descriptions of the preferred embodiments of the present invention and do not limit the scope of the present invention. Without departing from the spirit of the present invention's design, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention shall fall within the protection scope determined by the claims of the present invention.

Claims

1. An amphiphilic fluorinated organic polymer, characterized in that It has the general structure shown in formula (I): Formula (I); In formula (I), the value range of z is 11-230; m, x and y are the degree of polymerization, the value range of m is 5-200, the value range of x is 5-200, and the value range of y is 5-200.

2. The method for preparing an amphiphilic fluorinated organic polymer according to claim 1, characterized in that: The following steps are involved: A. In an oxygen-free and water-free environment, Compound A, structural formula: Compound B and catalyst 4-dimethylaminopyridine are mixed, tetrahydrofuran is used as the reaction solvent, and condensed and refluxed under heating and stirring. After the reaction is completed, the reaction liquid is separated and purified to obtain a structural formula The amphiphilic polymer PSMA-PEG; B. adding the amphiphilic polymer PSMA-PEG and N,N'-succinimidyl carbonate into anhydrous dichloromethane, then adding N,N-diisopropylethylamine, and stirring at room temperature for 6 h; C. adding excess amino-trifluoroaniline to the reaction system of step B, continuing stirring at room temperature for 48 h, after the reaction is completed, removing the solvent, dissolving the solid product in deionized water, dialyzing, and freeze-drying to obtain the amphiphilic fluorinated organic polymer; Among them, the value range of z is 11-230; m, x and y are the degree of polymerization, the value range of m is 5-200, the value range of x is 5-200, and the value range of y is 5-200.

3. The preparation method according to claim 2, characterized in that: The heating and stirring temperature in step A is 70° C. and the time is 24 h; the molar ratio of the amphiphilic polymer PSMA-PEG to N,N`-succinimidyl carbonate is 1:2~2:3; the molar ratio of the amphiphilic polymer PSMA-PEG to amino-trifluoroaniline is 1:1.2~1:

2.

4. Use of the amphiphilic fluorinated organic polymer as claimed in claim 1 as a carrier of a nano-diagnosis and treatment platform.

5. A nano-diagnosis and treatment platform, characterized in that: The amphiphilic fluorinated organic polymer and the fluorinated organic photosensitizer according to claim 1 are prepared by a nano-coprecipitation method; the coprecipitation method comprises the following steps: The amphiphilic fluorinated organic polymer and the fluorinated organic photosensitizer described in claim 1 are mixed in a tetrahydrofuran aqueous solution, and the solvent is removed under ultrasonication and a protective atmosphere to obtain water-soluble nanoparticles, which are the nano-diagnosis and treatment platform; The fluorine-containing organic photosensitizer has a general structure as shown in formula (II): Formula (II); In formula (II), R and R1 are independently selected from a branched or straight chain of C4 to C24.

6. Use of the nano-diagnosis and treatment platform as described in claim 5 in the preparation of tumor photothermal and / or photodynamic therapy agents.

7. Application of the nano-diagnosis and treatment platform as described in claim 5 as a photosensitizer for tumor photothermal and / or photodynamic therapy, and a tumor near-infrared zone II fluorescence imaging contrast agent.

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