Preparation method and application of nano diagnosis and treatment platform

Abstract

The invention discloses a preparation method and application of a nano diagnosis and treatment platform, and relates to the technical field of biological imaging and treatment. According to the nano diagnosis and treatment platform disclosed by the invention, the fluorinated amphiphilic organic polymer is used as a carrier and is tightly combined with the fluorinated organic semiconductor photosensitizer through a self-assembly process, so that nano particles with good hydrophobicity and biocompatibility are obtained; according to the nanoparticles, by virtue of unique fluorine-fluorine interaction among molecules and the oxygen capture capability of fluorine atoms, the yield of fluorescence quantum is improved, and the generation capability of < 1 > O2 is remarkably enhanced. The nano diagnosis and treatment platform disclosed by the invention not only shows excellent NIR-II FLI performance, but also has efficient PDT and photothermal conversion capability, and provides powerful technical support for realizing accurate and efficient tumor photodynamic / photothermal combined treatment.

Description

Technical Field

[0001] The present invention relates to the field of biological imaging and treatment technology, and in particular to a preparation method and application of a nano-diagnosis and treatment platform. Background Art

[0002] Compared with traditional surgery, radiotherapy and chemotherapy, optical diagnosis and treatment technology can detect cancer cells earlier. Through optical imaging technology, such as fluorescence imaging and photoacoustic imaging, three-dimensional visualization of tumor tissue can be achieved, providing doctors with detailed lesion information, so as 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 toxic side effects during treatment and significantly improving the quality of life of patients. These technologies are not only suitable for the treatment of superficial tumors, but also through the use of a variety of innovative methods, 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, etc., constantly breaking through limitations and expanding into the field of deep tumor treatment. With the continuous advancement and deep integration of nanoscience and biotechnology, a series of more accurate, efficient and low-side effect cancer treatment solutions have been developed one after another.

[0003] Near infrared II (NIR-II, 1000-1700nm) fluorescence imaging technology (NIR-II FLI), with its superior penetration depth, high-resolution imaging capabilities, and real-time dynamic monitoring of internal structure and functional changes of organisms, is gradually becoming a cutting-edge tool for biological visualization. In the process of promoting the clinical transformation of NIR-II FLI, the development of high brightness (high fluorescence quantum yield) Photosensitizers that combine molar extinction coefficient ε), long wavelength, high photostability and biocompatibility are crucial.

[0004] As an innovative cancer treatment, the core principle of PDT is to use photosensitizers to convert light energy into reactive oxygen species (ROS) to induce cell apoptosis and tissue damage. 1 O 2 However, its efficacy is limited by the hypoxic environment inside the tumor. To break through this bottleneck, researchers are actively exploring various strategies to enhance the therapeutic effect of PDT under hypoxic conditions. For example, hyperbaric oxygen therapy or the development of nanocarriers for oxygen delivery can increase the oxygen content in the tumor to increase 1 O 2The development of oxygen-independent type I photosensitizers or dual-mechanism photosensitizers combining type I and type II mechanisms to effectively kill tumor cells in anoxic environments; at the same time, the performance of photosensitizers is continuously optimized, such as increasing fluorescence quantum yield and molar extinction coefficient, improving photostability and enhancing biocompatibility, in order to enhance the overall efficacy of PDT. In addition, PDT is also combined with other treatments such as PTT, chemotherapy, radiotherapy and immunotherapy, using synergistic effects to further enhance the therapeutic effect and reduce side effects.

[0005] Fluorination modification has shown significant advantages in the biomedical field, especially in reducing physiological toxicity and improving therapeutic effects. Its unique advantages come from the chemical properties of fluorine atoms, namely its larger electronegativity and smaller atomic radius, which makes fluorinated compounds more stable and less reactive in vivo, thereby effectively reducing toxic side effects on normal cells. At the same time, fluorination can also stabilize the electronic structure of fluorescent materials, reduce non-radiative transitions, and thus improve fluorescence quantum yield and optical properties. In addition, fluorinated polymers have excellent oxygen capture capabilities, so the fluorination strategy is expected to improve the hypoxic environment inside the tumor and provide more oxygen sources for PDT, thereby significantly improving the therapeutic effect. Amphiphilic polymers are widely used in drug delivery systems due to their unique structure (containing both hydrophilic and hydrophobic parts). Fluorination modification, as an effective terminal modification method, can not only enhance the stability and biocompatibility of polymer carriers, but also give them new functional properties. These improvements make fluorinated amphiphilic polymer carriers show broader application potential and prospects in the field of advanced photodiagnosis and treatment. 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 object, the present invention provides the following solutions:

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

[0009]

[0010] 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.

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

[0012] The second object of the present invention is to provide a method for preparing the above-mentioned amphiphilic fluorine-containing organic polymer, comprising the following steps:

[0013] A. In an oxygen-free and water-free environment, The commercial compound A has the structural formula

[0014] The commercial compound B and the catalyst 4-dimethylaminopyridine (DMAP) are mixed, tetrahydrofuran (THF) 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 compound with the structural formula The amphiphilic polymer PSMA-PEG;

[0015] B. adding the amphiphilic polymer PSMA-PEG and N,N'-succinimidyl carbonate (DSC) into anhydrous dichloromethane (DCM), then adding N,N-diisopropylethylamine (DIPEA), and stirring at room temperature for 6 hours;

[0016] C. Add excess amino-trifluoroaniline (NH 2 -ph-3F), and continue stirring at room temperature for 48 hours. After the reaction is completed, remove the solvent, dissolve the solid product in deionized water, dialyze, and freeze-dry 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, the heating and stirring temperature in step A is 70° C. and the time is 24 hours; the molar ratio of the amphiphilic polymer PSMA-PEG to N,N`-succinimidyl carbonate is 1:2 to 2:3; the molar ratio of the amphiphilic polymer PSMA-PEG to amino-trifluoroaniline is 1:1.2 to 1:2.

[0020] The third object of the present invention is to provide a fluorine-containing organic photosensitizer 4F having a general structure as shown in formula (II):

[0021]

[0022] In formula (II), R and R 1 Each independently selected from a branched or straight chain of C4 to C24.

[0023] The fourth object of the present invention is to provide a method for preparing the above-mentioned fluorine-containing organic photosensitizer.

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

[0025] Among them, R and R 1 Each independently selected from a branched or straight chain of C4 to C24.

[0026] More specifically, the following steps are included:

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

[0028] Among them, R and R 1 Each independently selected from a branched or straight chain of C4 to C24.

[0029] The fifth object of the present invention is to provide the application of the above-mentioned amphiphilic fluorinated organic polymer as a carrier of a nano-diagnosis and treatment platform.

[0030] The sixth object of the present invention is to provide the application of the above-mentioned fluorine-containing organic photosensitizer as a photosensitizer for a nano-diagnosis and treatment platform.

[0031] The seventh object of the present invention is to provide a nano-diagnosis and treatment platform, which is prepared by the above-mentioned amphiphilic fluorinated organic polymer and fluorinated organic photosensitizer through a nano-coprecipitation method; the co-precipitation method comprises the following steps: mixing the above-mentioned amphiphilic fluorinated organic polymer and fluorinated organic photosensitizer in a tetrahydrofuran aqueous solution, ultrasonicating, removing the solvent under a protective atmosphere to obtain water-soluble nanoparticles, namely the nano-diagnosis 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 objective of the present invention is to provide the application of the above-mentioned nano-diagnosis and treatment platform in the preparation of tumor photothermal and / or photodynamic therapy reagents.

[0034] The ninth objective of the present invention is to provide the application of the above-mentioned nano-diagnosis and treatment platform as a photosensitizer for tumor photothermal and / or photodynamic therapy, and a tumor near-infrared second-zone fluorescence imaging (NIR-II FLI) contrast agent.

[0035] The present invention successfully constructs a nano-diagnosis and treatment platform by using the prepared amphiphilic organic polymer rich in fluorine atoms as a carrier and co-assembling with fluorinated organic semiconductor small molecules with excellent light absorption ability. The nano-diagnosis and treatment platform greatly improves the fluorescence quantum yield (QY) by using the fluorine-fluorine interaction between molecules, and has a stronger oxygen capture ability due to its rich fluorine atoms, effectively enhancing 1 O 2 The production of.

[0036] The nano-diagnosis and treatment platform of the present invention is to encapsulate the fluorine-containing photosensitizer in the NH 2 -ph-3F modified fluorinated amphiphilic organic polymer (PSMA and NH 2 The nano-theranostic platform has fluorination-enhanced NIR-II FLI and PDT effects. The preparation process of the nano-theranostic platform is to first modify amino-polyethylene glycol (NH4+) on isopropylbenzene-terminated poly(styrene-co-maleic anhydride) (PSMA) by amidation reaction. 2 -PEG), thereby obtaining an amphiphilic polymer carrier. Subsequently, through further amidation reaction, the fluorine source molecule amino-trifluoroaniline was introduced into the PSMA-PEG polymer backbone to construct an amphiphilic polymer nanocarrier PSMA-3F containing high-density fluorine. Next, the above-mentioned fluorine-containing organic photosensitizer (abbreviated as 4F) and the polymer nanocarrier PSMA-3F were combined by nanocoprecipitation in a mixed solvent of water and tetrahydrofuran to prepare water-soluble nanoparticles FF NPs. These nanoparticles exhibit fluorination-enhanced NIR-IIFLI performance in water, and under 808nm laser irradiation, they exhibit enhanced singlet oxygen ( 1 O 2 ) generation capacity and good photothermal conversion efficiency. Therefore, this nano-diagnosis and treatment platform is expected to achieve synergistic treatment of tumors with PDT and photothermal therapy (PTT) under the guidance of NIR-II FLI.

[0037] The present invention provides an application of the above-mentioned nano-diagnosis and treatment platform as a tumor NIR-II FLI contrast agent. Specifically, a near-infrared two-window fluorescence imaging method is as follows: a certain concentration of FF NPs aqueous solution is prepared, placed in a small centrifuge tube, and excited with an 808nm laser under a NIR-II fluorescence imager to obtain a NIR-II FLI image.

[0038] The present invention provides the application of the above-mentioned nano-diagnosis and treatment platform as a photosensitizer for photothermal and / or photodynamic therapy of tumors. Specifically, in the application of photodynamic therapy agents, the following testing steps are included:

[0039] A certain concentration of FF NPs aqueous solution was added to the cuvette, and then 1 O 2 Indicator (SOSG, 1-10 μM). Use a laser with set optical power density (808 nm, 0.1, 0.25, 0.5, 0.75, 1 W cm -2 ) The sample was illuminated intermittently, and the fluorescence intensity at 521nm in the fluorescence spectrum under different illumination times was recorded by a spectrometer, and a curve changing with illumination time was drawn. At the same time, a blank group (only adding the same concentration of SOSG) and a control group (using PF NPs nanowater solution plus the same concentration of SOSG) were set up, and the experimental operation and data were recorded according to the same steps.

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

[0041] The small centrifuge tubes containing deionized water (blank group) and the small centrifuge tubes containing the same concentration of FF NPs or PF NPs aqueous solution were illuminated by a laser with a certain light power density, and then cooled to room temperature. At the same time, an infrared thermographic camera was used to record the trend of the solution temperature in the two centrifuge tubes over time.

[0042] The PF NPs aqueous solution is prepared by directly encapsulating the fluorine-containing organic photosensitizer 4F with the fluorine-free amphiphilic organic polymer PSMA-PEG.

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

[0044] The core of the nano-diagnosis and treatment platform of the present invention is to use fluorinated amphiphilic organic polymers as carriers, which are tightly combined with fluorinated organic semiconductor photosensitizers through a self-assembly process to obtain nanoparticles with good hydrophobicity and biocompatibility. These nanoparticles achieve an increase in fluorescence quantum yield and significantly enhance the 1 O 2 generating capacity.

[0045] The nano-diagnosis and treatment platform of the present invention not only exhibits superior NIR-II FLI performance, but also has efficient PDT and photothermal conversion capabilities, providing strong technical support for achieving accurate and efficient combined photodynamic / photothermal treatment of tumors.

[0046] The fluorination-enhanced nano-diagnostic and therapeutic platform of the present invention, with its excellent imaging quality and significant therapeutic effect, can achieve synergistic treatment of PDT and PTT under the precise guidance of NIR-II FLI, opening up a new approach for the treatment of diseases such as inflammation and tumors, and bringing patients safer and more effective treatment options. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

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

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

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

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

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

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

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

[0055] Figure 8 The 10 μM aqueous solutions of FF NPs and PF NPs were subjected to the same light power density (808 nm, 0.5 W / cm 2 ) When the laser is irradiated, each 1 O 2 The production situation.

[0056] Fig. 9 The 12 μM aqueous solutions of FF NPs and PF NPs were subjected to the same light power density (808 nm, 0.5 W / cm 2 ) When irradiated by laser, the photothermal temperature rises and falls.

[0057] Fig.10 Confocal fluorescence imaging of 4T1 cells' cellular uptake of FF NPs or PF NPs (a) and quantitative analysis results of their fluorescence intensity (b).

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

[0059] Fig.12 NIR-II tumor fluorescence imaging of living mice after tail vein injection of FF NPs (a) and the curve of fluorescence intensity change over time at the tumor site (b). DETAILED DESCRIPTION

[0060] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0061] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0062] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may 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 associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0063] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0064] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

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

[0066] The following is a specific description with reference to the embodiments.

[0067] Example 1

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

[0069]

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

[0071] Figure 2 The fluorine-containing organic photosensitizer 4F prepared in Example 1 1 H NMR spectrum, Figure 3 The following is the MALDI-TOF mass spectrum.

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

[0073]

[0074] Poly(styrene-co-maleic anhydride) (isopropylbenzene capped, 0.1mmol, Mn~1900), amino-polyethylene glycol (Mn~5000, 0.3mmol) and catalyst DMAP were added to the reaction flask. Subsequently, sufficient THF was added to the reaction flask as the reaction solvent. Stirring and heating to 70°C, condensing and refluxed to prevent solvent evaporation. The reaction was carried out under this condition for 24 hours. After the reaction was completed, the reaction mixture was cooled to room temperature and precipitated with ether. Subsequently, the precipitate was re-dissolved with deionized water and freeze-dried to obtain the amphiphilic polymer PSMA-PEG (Mn~6974kDa).

[0075] (3)PSMA-3F synthesis:

[0076]

[0077] The product PSMA-PEG (0.1 mmol) and DSC (0.2 mmol) were added to a reaction flask, and sufficient anhydrous DCM was added as a reaction solvent, and then a small amount of DIPEA was slowly added as a catalyst. The reaction mixture was stirred for 6 hours at room temperature. Then, excess NH 2 -ph-3F (0.2 mmol) was added into the reaction flask which had been stirred for 6 hours, and the stirring was continued at room temperature for 48 hours to introduce a fluorine source into the polymer skeleton.

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

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

[0080] (3) Preparation of nano-diagnostic and therapeutic platform: The PSMA-3F and 4F prepared above (mass ratio 5:1) were placed in a binary solution of THF and deionized water (volume ratio 1:9) using a nano-coprecipitation method, and nanoparticles FFNPs were obtained under ultrasound. The THF was then evaporated and removed under nitrogen protection.

[0081] Comparative Example 1

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

[0083] By using a 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 ultrasound, and then THF was evaporated and removed under nitrogen protection.

[0084] Performance Testing:

[0085] (1) UV absorption and fluorescence emission test of FF NPs and PF NPs. The specific steps are as follows:

[0086] 3 mL of FF NPs and PF NPs aqueous solutions of the same concentration were placed in two cuvettes, and then placed in a UV spectrophotometer to collect UV absorption spectra. Next, in the same way, the two samples were placed in a fluorescence spectrometer to collect their respective emission spectra. The results are shown in Figure 2. 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 exhibit strong absorption 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 their emission tail bands extend to 1300nm in aqueous solution, which can effectively avoid fluorescence interference in in vivo imaging and is conducive to high-contrast NIR-II FLI of deep tissues. It is worth noting that at the same concentration, FF NPs exhibited significantly higher fluorescence intensity than PF NPs, which clearly indicates the significant enhancement effect of fluorination on fluorescence properties.

[0088] (2) NIR-II FLI test of FF NPs and PF NPs. The specific steps are as follows:

[0089] 300 μL of FF NPs and PF NPs aqueous solutions with the same concentration were placed in two 0.5 mL centrifuge tubes, and then placed in the NIR-II fluorescence imager and irradiated with 808 nm laser at a power density of 0.5 W / cm 2 , 1000nm long pass filter, and fluorescence image acquisition. Figure 7 shown.

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

[0091] (3) FF NPs and PF NPs 1 O 2 Generate capability test, the specific steps are as follows:

[0092] SOSG (Singlet Oxygen Sensor Green) singlet oxygen probe is used to detect 1 O 2Generation. 10 μL SOSG (working concentration: 5 μM) was mixed with FF NPs or PF NPs aqueous solution (working concentration: 10 μM). 10 μL SOSG was mixed with 3 mL deionized water as a control group. Afterwards, the mixture was exposed to 808 nm laser (0.5 W / cm 2 ), and the fluorescence intensity at about 521 nm was recorded in a fluorescence spectrometer (excitation wavelength: 488 nm) for 30 seconds each time. Figure 8 shown.

[0093] Figure 8 FF NPs or PF NPs in water phase with 1 O 2 After mixing, the fluorescence intensity at 521nm wavelength changes with illumination time. Figure 8 It can be seen that when only SOSG exists, the fluorescence signal intensity of the solution has almost no significant change after illumination. In contrast, when 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 when FF NPs exist. This shows that both FF NPs and PF NPs have good 1 O 2 The FF NPs showed better performance in this regard, which may be because the fluorinated FF NPs enhanced their ability to capture oxygen.

[0094] (4) Photothermal conversion efficiency test of FF NPs and PF NPs. The specific steps are as follows:

[0095] The photothermal conversion efficiency of FF NPs and PF NPs was calculated. 300 μL of FF NPs and PF NPs aqueous solution with the same concentration was irradiated with 808 nm laser (0.5 W / cm 2 ) for 5 min, and monitor the temperature changes during the illumination period. Subsequently, turn off the laser, cool the solution to room temperature, and continue to monitor the temperature changes during the cooling process. During this process, the solution temperature changes were recorded every 30 seconds. In addition, 300 μL of deionized water was set as a blank control group. The results are shown in Fig. 9 shown.

[0096] according to Fig. 9 The obtained data can be used to calculate the photothermal conversion efficiency of FF NPs and PF NPs, respectively: PCE FF NPs =52.8% and PCE PF NPs =51.6%. This indicates that FF NPs and PF NPs have excellent photothermal properties and show potential for in vivo tumor treatment.

[0097] (5) Cellular uptake test of FF NPs or PF NPs. The specific steps are as follows:

[0098] Mouse breast cancer cells (4T1) were inoculated in a confocal microscope dish and incubated for at least 24 hours. FF NPs or PF NPs (20 μM) were then added and incubated for 9 hours. At the same time, a blank group without any material was set up and recorded as the Control group. After the incubation, the cells were washed 3 times with PBS. 100 μL of 1×Hoechst33342 nuclear stain was added to each well and incubated at room temperature in the dark for 30 minutes. The stain was then removed and washed 3 times with PBS. Finally, the cells were observed using a confocal microscope (Zeiss) in sequential scanning mode. FF NPs or PF NPs (λex=565nm, λem=780nm), Hoechst33342 (λex=346nm, λem=461nm). The results are shown in Fig.10 shown.

[0099] Fig.10 The results of confocal imaging confirmed that FF NPs or PF NPs could be well taken up by cells, and FF NPs in 4T1 cells showed stronger fluorescence after 9 hours, which may be related to the longer retention of FF NPs in cells.

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

[0101] The CCK-8 cell viability test kit was used. 4T1 cells were cultured with DMEM complete medium (containing 10% FBS, 1% penicillin / streptomycin) in a humidified incubator (37°C, 5% CO 2 , 20% O 2 ) were cultured. Then, the cells in the logarithmic growth phase were harvested and cultured at 1×10 4 The cells were seeded in a 96-well plate at a density of 10 cells / mL 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, they were exposed to 808 nm laser irradiation for 5 min (0.5 W / cm 2), recorded as FF NPs+L or PF NPs+L, respectively. At the same time, cells incubated with FF NPs or PF NPs without laser irradiation were set as dark control groups. After further incubation for 18 hours, the culture medium was removed and washed three times with PBS. The cells were then incubated with fresh serum-free basic medium containing 10% CCK-8 in the dark for 30 minutes. Finally, the absorbance of the product was measured at a wavelength of 450nm using an enzyme reader. The results are expressed as the survival percentage of cells after different treatments relative to the control group cells without any treatment. The results are shown in Fig.11 shown.

[0102] Fig.11 This is an analysis test of the photodynamic / photothermal synergistic killing ability of FF NPs or PF NPs on 4T1 cells. It can be seen that with the increase in the concentration of nanomaterials, the cell viability in the non-illuminated group did not decrease. On the contrary, whether incubated with FF NPs or PF NPs, under illumination, the viability of 4T1 cells decreased with the increase in material concentration. When the concentration of both was 15μM, the cell killing effect was obvious, and FF NPs showed a stronger cell killing ability. According to the aforementioned in vitro performance test, it can be analyzed that when the photothermal conversion efficiency is basically the same, that is, the photothermal effect is almost the same, FF NPs are stronger. 1 O 2 The fluorinated FF NPs have excellent synergistic phototherapy effects and show better tumor killing potential.

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

[0104] In order to study the feasibility of using FF NPs for in vivo fluorescence imaging to guide tumor therapy, a solid tumor model was established by subcutaneous injection of 4T1 cells. After tail vein injection of FF NPs aqueous solution, fluorescence imaging images were collected at different time points (0h, 1h, 3h, 6h, 12h, 24h, 48h) after injection of the mouse tumor site. The results are shown in Fig.12 shown.

[0105] Due to the enhanced permeability and retention (EPR) effect of tumors, Fig.12It can be seen that after the tail vein injection of FF NPs, the mouse tumor site quickly showed bright fluorescence, and the fluorescence signal intensity reached a peak within 3 hours. Although it weakened slightly afterwards, it could still maintain a relatively obvious fluorescence brightness for a long time. This result shows that FF NPs can quickly penetrate the tumor blood vessel wall during blood circulation and accumulate in the tumor tissue, and show excellent tumor retention performance. This indicates that these nanoparticles have great potential as fluorescent imaging probes to guide photodynamic / photothermal therapy of deep tumors.

[0106] In summary, the cancer nano-diagnosis and treatment platform for fluorination-enhanced NIR-II FLI and PDT efficiency provided by the present invention has simple synthesis steps, simple separation and purification operations, and has obvious fluorescence enhancement and improved 1 O 2 It has high photothermal conversion performance, strong killing ability against breast cancer cells, can be used for in vivo vascular imaging, and has application potential in constructing anticancer drugs with efficient NIR-II FLI-guided photodynamic / photothermal synergistic therapeutic effects.

[0107] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all 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): 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 to anhydrous dichloromethane, then adding N,N-diisopropylethylamine, and stirring at room temperature for 6 hours; C. adding excess amino-trifluoroaniline to the reaction system of step B, continuing stirring at room temperature for 48 hours, after the reaction is completed, removing the solvent, dissolving the solid product in deionized water, dialyzing, and freeze-drying to obtain the amphiphilic fluorine-containing 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 hours; the molar ratio of the amphiphilic polymer PSMA-PEG to N,N`-succinimidyl carbonate is 1:2 to 2:3; the molar ratio of the amphiphilic polymer PSMA-PEG to amino-trifluoroaniline is 1:1.2 to 1:

2.

4. A fluorine-containing organic photosensitizer 4F, characterized in that: It has the general structure shown in formula (II): In formula (II), R and R1 are independently selected from branched or straight chains of C4 to C24.

5. The method for preparing a fluorine-containing organic photosensitizer according to claim 4, characterized in that: The following steps are involved: The structural formula is The compound 1 and the structural formula are Compound 2 is dissolved in chloroform under an argon atmosphere, and then pyridine is added, and the reaction mixture is heated to reflux at 55° C. for 16 hours. After the reaction is completed, the reaction mixture is cooled to room temperature, poured into methanol to precipitate a crude product, which is collected by filtration and purified to obtain the fluorine-containing organic photosensitizer; Wherein, R and R1 are independently selected from branched or straight chains of C4 to C24.

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

7. Use of the fluorine-containing organic photosensitizer as claimed in claim 4 as a photosensitizer for a nano-diagnosis and treatment platform.

8. A nano-diagnosis and treatment platform, characterized in that: The amphiphilic fluorine-containing organic polymer according to claim 1 and the fluorine-containing organic photosensitizer according to claim 3 are prepared by nano-coprecipitation method; the coprecipitation method comprises the following steps: The amphiphilic fluorinated organic polymer described in claim 1 and the fluorinated organic photosensitizer described in claim 4 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.

9. Use of the nano-diagnosis and treatment platform as described in claim 8 in the preparation of tumor photothermal and / or photodynamic therapy agents.

10. Application of the nano-diagnosis and treatment platform as claimed in claim 8 as a photosensitizer for photothermal and / or photodynamic therapy of tumors, and as a contrast agent for tumor near-infrared zone II fluorescence imaging.

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