Near-infrared light driven fluorescence imaging nano composite material, preparation method and application in collaborative treatment
By depositing sheet-like manganese dioxide on the surface of UCNP upconverting the luminescent nanoparticles and loading indocyanine green to form a flower-like or honeycomb-like core-shell structure, a multimodal synergistic treatment driven by near-infrared light is achieved, and the problem of insufficient treatment effect in the prior art is solved, which improves tumor killing effect and reduces side effects.
Patent Information
- Application Number
- CN202510528860.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-25
- Publication Date
- 2025-07-11
AI Technical Summary
Existing nanomaterials are difficult to achieve multimodal synergistic treatment driven by near-infrared light, especially the lack of synergistic effects of photodynamic therapy, photothermal therapy and chemodynamic therapy, which limits the improvement of tumor killing effects and the reduction of side effects.
By constructing the upconversion luminescent nanoparticle UCNP as the core, the flaky manganese dioxide is deposited on the surface to form a flower-like or honeycomb-like core-shell structure, and loading indocyanine green molecules to form a CDT/PTT/PDT multimodal complementary mechanism, and synergistic treatment is achieved using multi-enzyme catalysis and near-infrared light mediation.
It significantly enhances the tumor killing effect, reduces side effects, and realizes the synergy between high-resolution fluorescence imaging, photothermal therapy, photodynamic therapy and chemokinetic therapy, providing technical ideas for integrating early clinical diagnosis and multimodal therapy.
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Figure CN120285238A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the fields of nanomaterials and medical technologies, and particularly relates to a near-infrared light-driven fluorescence imaging nanocomposite material, a preparation method thereof, and an application in multimodal synergistic therapy. Background Art
[0002] Cancer is one of the major threats to global health. At present, most cancer patients diagnosed clinically are in the middle and late stages, and the best treatment time has been missed. Traditional surgical treatment has high risks and a five-year survival rate of less than 50%. Therefore, early and accurate monitoring and efficient treatment of cancer have become the core challenges in current research.
[0003] Chemotherapy and radiotherapy are the two major means of treating cancer clinically at present, but neither of them can completely ablate tumors. In addition to the commonly used chemotherapy and radiotherapy treatment means in clinical practice, emerging tumor treatment technologies mainly include photothermal therapy, photodynamic therapy, and chemodynamic therapy. Indocyanine green (ICG) is a near-infrared (NIR) fluorescent dye with strong absorption in the wavelength range of 700 - 900 nm. Under the irradiation of near-infrared light, it can convert light energy into heat energy and generate a large amount of reactive oxygen species (ROS), which can be further used for photothermal therapy (PTT) and photodynamic therapy (PDT). Chemodynamic therapy (CDT) is a new cancer treatment strategy based on the tumor microenvironment and Fenton chemical reactions. Its core mechanism is to generate highly toxic hydroxyl radicals (•OH) by catalyzing endogenous hydrogen peroxide (H2O2) in tumor cells, thereby selectively killing cancer cells. Compared with traditional treatment methods, CDT has significant advantages such as less side effects on normal tissues and being less likely to induce drug resistance. In CDT, flower-like manganese dioxide (MnO2) nanostructures, as a multifunctional catalytic platform, exhibit characteristics superior to traditional iron-based catalysts, that is, MnO2 nanoflowers have pH-responsive multi-enzyme mimicking activities, including peroxidase, oxidase, catalase, and superoxide dismutase, which can significantly enhance the generation of toxic free radicals through logical catalysis, and the O2 generated during the catalytic process and the consumption of reduced glutathione contribute to further enhancing the therapeutic effect of PDT.
[0004] As an emerging material, nanomaterials have significant advantages in the diagnosis and treatment of tumors due to their unique properties. Among them, upconversion nanoparticles (UCNP) have the advantages of high optical and chemical stability, low biotoxicity, high luminescence intensity, narrow emission band and easy regulation, etc., and have attracted the favor of researchers in the research of precise tumor diagnosis and efficient treatment. The excitation light of upconversion nanoparticles is usually located in the near-infrared light region, which has a relatively deep tissue penetration depth, can effectively avoid the autofluorescence of organisms, and has almost no damage to biological tissues. Therefore, upconversion nanoparticles have good application potential in biological marker detection, disease diagnosis and treatment, etc.
[0005] However, how to skillfully combine the above various nanomaterials and components to enable them to simultaneously support the synergistic diagnosis and treatment of photothermal therapy, photodynamic therapy and chemodynamic therapy is a technical problem in this field.
[0006] In the prior art, a preparation method and application of an open hollow manganese dioxide-based nanozyme composite material disclosed in CN115770295A cover a manganese dioxide layer on the surface of silica nanoparticles through redox reaction, use sodium hydroxide solution as an etching agent for etching, load the open hollow manganese dioxide nanoparticles with ce6 photosensitizer, cover a polydopamine layer on the surface of the open hollow manganese dioxide nanoparticles loaded with ce6 photosensitizer, and simultaneously adsorb glucose oxidase through electrostatic adsorption and π-π stacking adsorption, so as to improve the tumor killing ability through the combination of tumor photodynamic, photothermal therapy, chemodynamic therapy and starvation therapy technologies. However, in practical applications, it is found that this composite material uses open hollow manganese dioxide nanoparticles to load the photosensitizer chlorin e6, which can generate reactive oxygen species through photodynamic therapy to kill tumor cells. Its open hollow manganese dioxide has a large specific surface area and has a certain drug loading capacity. The photosensitizer chlorin e6 is loaded in the hollow manganese dioxide to play a role. However, this diagnostic and therapeutic agent has certain limitations. For example, it is difficult to achieve multimodal synergistic therapy (including photodynamic therapy, photothermal therapy and chemodynamic therapy) of near-infrared light-driven upconversion imaging, thus limiting its diagnostic and therapeutic means and effects, and unable to further improve the comprehensive effect of the material and reduce side effects.
[0007] In the prior art, a nano-material with a three-dimensional flower-like structure capable of loading drugs, its preparation method and application disclosed in CN110856748B is a multi-layer core-shell structure, which includes a core, an intermediate layer, and a shell layer. The core is made of rare earth nano-materials, the intermediate layer is a mesoporous silica layer, and the shell layer is a sodium sulfide layer. This three-dimensional flower-like nano-structure has good drug-loading ability, and drugs can be loaded on it to form a complex, and tumor cells can be killed through photothermal therapy, chemotherapy and other methods. The nickel sulfide of the three-dimensional flower-like nano-structure of this composite material is used as a photothermal material, which can complete photothermal conversion under the irradiation of an 808 nm laser, and then kill cancer cells. Its three-dimensional flower-like structure is used for drug loading. However, this diagnostic agent also has certain limitations. For example, the flower-like nickel sulfide structure can only undertake the function of photothermal therapy and cannot improve the synergistic treatment effect (such as generating oxygen, assisting in consuming reducing substances in cancer cells, etc.), thus limiting the improvement of the comprehensive treatment effect of the material.
[0008] In the prior art, CN117618558A discloses a functional material for a diagnostic and therapeutic integrated nano-composite system, its preparation method and application. The functional material therein uses upconversion luminescent nanoparticles UCNP as the core, and dopamine monomers are coated on the surface of the core to form a mesoporous polydopamine layer. Then, arginine is combined with the mesoporous polydopamine, and indocyanine green is loaded into the mesoporous polydopamine layer. The obtained nano-composite system functional material modified by indocyanine green and arginine, a natural biomolecule, can be applied to biomedical fields such as three-modal synergistic therapy of cancer cell photodynamic therapy, photothermal therapy and NO gas therapy mediated by fluorescence imaging. Specifically, the upconversion nanoparticles serve as an imaging center and can perform optical imaging under the excitation of a 980 nm laser; the coating of mesoporous dopamine endows the system with the ability to load drugs; indocyanine green will generate reactive oxygen species under the irradiation of an 808 nm laser to achieve photodynamic therapy, and at the same time, a large amount of heat will be generated to achieve photothermal therapy; L-arginine, as a NO gas therapy reagent, can be activated by the reactive oxygen species generated by indocyanine green to generate NO, realizing the gas therapy function. However, the multi-modal synergistic treatment performance of the above-mentioned diagnostic and therapeutic integrated reagent still has certain deficiencies. Mesoporous dopamine only acts as a modified coating layer in it, providing more reaction sites for the system, but it cannot play a corresponding therapeutic role. Moreover, compared with the three-dimensional flower-like structure, its drug-loading ability is slightly weaker. Mesoporous dopamine also cannot help other therapeutic drugs play their roles, and the NO gas therapy effect of L-arginine will also weaken the photodynamic therapy effect brought by indocyanine green. The various treatment modes cannot promote each other, limiting the further improvement of the treatment effect of the material.
[0009] In view of the deficiencies of the above-mentioned existing nano-materials, it is particularly important to further research and improve the multi-modal synergistic treatment effect. Summary of the Invention
[0010] The object of the present invention is to propose a newly conceived near-infrared light-driven fluorescence imaging nanocomposite and a preparation method in view of the limitations existing in the existing treatment technologies, especially the problem of being difficult to improve the comprehensive performance of nanomaterials and enhance the tumor killing effect. Through unique component, process design and microstructure optimization, it can simultaneously support near-infrared light-driven upconversion fluorescence imaging and its mediated multi-modal synergistic treatment of cancer cells (such as photodynamic therapy, photothermal therapy and chemodynamic therapy, etc.), and form a CDT / PTT / PDT multi-modal complementary mechanism under the mediation of multi-enzyme catalysis and near-infrared light, significantly enhancing the diagnosis and treatment effect and reducing side effects, and providing new technical ideas and strategies for the field of clinical early diagnosis and multi-modal treatment integration.
[0011] To achieve the above object, the technical solution adopted by the present invention is as follows: A near-infrared light-driven fluorescence imaging nanocomposite, which takes upconversion luminescent nanoparticles UCNP as the core, deposits sheet-like manganese dioxide on the surface of the core to form a flower-like or honeycomb-like core-shell structure, then modifies the manganese dioxide with amino groups, and further loads indocyanine green molecules. The obtained upconversion diagnosis and treatment integrated nanocomposite modified by indocyanine green molecules and flower-like or honeycomb-like manganese dioxide forms a CDT / PTT / PDT multi-modal complementary mechanism under the mediation of multi-enzyme catalysis and near-infrared light.
[0012] The upconversion luminescent nanoparticles UCNP are upconversion luminescent nanoparticles coated with oleic acid. The oleic acid molecules on the surface of UCNP react with potassium permanganate through redox reaction to generate sheet-like manganese dioxide, and the manganese dioxide lamellae further self-assemble to form a flower-like structure. Amino groups are loaded on the surface of manganese dioxide through the hydrolysis and polycondensation reaction of 3-aminopropyltriethoxysilane in a toluene environment, and the obtained amino-functionalized flower-like or honeycomb-like manganese dioxide-coated UCNP is obtained.
[0013] The amino groups modified on the surface of manganese dioxide are condensed or nucleophilically substituted with indocyanine green to load indocyanine green on the surface of flower-like or honeycomb-like manganese dioxide, and a upconversion diagnosis and treatment integrated nanocomposite functional material modified by indocyanine green and flower-like manganese dioxide is obtained.
[0014] A preparation method of a near-infrared light-driven fluorescence imaging nanocomposite, comprising the following steps: S1. Preparation of the first dispersion: Prepare cyclohexane dispersion, potassium permanganate aqueous solution, and oleic acid containing UCNP respectively; add oleic acid to the cyclohexane dispersion, ultrasonicate at room temperature and then stir for a period of time to evaporate cyclohexane, obtaining an oleic acid dispersion containing UCNP; then prepare potassium permanganate, dissolve it in a certain amount of deionized water to obtain a potassium permanganate aqueous solution, mix this potassium permanganate aqueous solution with the oleic acid dispersion containing UCNP, stir at room temperature for a set time, ultrasonically disperse and then centrifuge, take the solid and disperse it in deionized water to prepare the first dispersion; S2. Preparation of the second dispersion: Prepare 3-aminopropyltriethoxysilane, freeze-dry the first dispersion, disperse the obtained powder in a certain amount of toluene solution, ultrasonicate at room temperature and then stir for a period of time; then add an appropriate amount of 3-aminopropyltriethoxysilane to the above stirred mixture, raise the temperature and stir and reflux at a set temperature for a set time, centrifuge and take the lower-layer solid and disperse it in deionized water to prepare the second dispersion; S3. Co-modification: Dissolve indocyanine green in a certain amount of deionized water to obtain an indocyanine green aqueous solution, mix it with the second dispersion in a specified ratio, stir in the dark at room temperature until the set time, centrifuge and take the lower-layer solid and disperse it in deionized water, so that indocyanine green undergoes a condensation reaction or nucleophilic substitution reaction with the amino group on manganese dioxide to form a covalent bond, and then load indocyanine green onto manganese dioxide, obtaining a near-infrared light-driven fluorescence imaging nano-composite functional material co-modified by indocyanine green and flower-like or honeycomb-like manganese dioxide, that is, a near-infrared light-driven fluorescence imaging nano-composite material.
[0015] Application of the above-mentioned near-infrared light-driven fluorescence imaging nano-composite material: Use it as a near-infrared light-driven fluorescence imaging contrast agent.
[0016] Application of the above-mentioned near-infrared light-driven fluorescence imaging nano-composite material: Use it as a photothermal therapy reagent, a photodynamic therapy reagent, and a treatment reagent for triple-mode synergistic therapy of chemodynamic under near-infrared light excitation to achieve triple-mode synergistic therapy under near-infrared light excitation.
[0017] Application of the above-mentioned near-infrared light-driven fluorescence imaging nano-composite material: Use it as a treatment reagent for multi-enzyme catalysis and near-infrared light-mediated CDT / PTT / PDT multi-modal complementary synergistic therapy.
[0018] The near-infrared light-driven fluorescence imaging nano-composite material, its preparation method and application provided by the present invention have at least the following beneficial effects: 1. The focus of the present invention is to improve the comprehensive performance of nanomaterials by constructing a clever microstructure, enhance the tumor killing effect and reduce side effects. Specifically, through unique component, process design and microstructure optimization (flower-like or honeycomb-like core-shell structure), the nanocomposite can simultaneously support near-infrared light-driven upconversion fluorescence imaging and its mediated multimodal synergistic therapy for cancer cells (such as photodynamic therapy, photothermal therapy and chemodynamic therapy). Under the mediation of multi-enzyme catalysis and near-infrared light, a CDT / PTT / PDT multimodal complementary mechanism is formed, significantly enhancing the diagnosis and treatment effect and reducing side effects, providing new technical ideas and strategies for the field of clinical early diagnosis and integrated multimodal treatment; in particular, the fluorescence imaging-mediated triple-modal synergistic therapy for cancer cells is a cancer treatment method that combines fluorescence imaging, photodynamic therapy (PDT), photothermal therapy (PTT) and chemodynamic therapy (CDT). This treatment method utilizes the complementary advantages of different treatment modes, which can significantly improve the treatment effect and reduce side effects.
[0019] 2. The nanocomposite for near-infrared light-driven fluorescence imaging and triple-modal synergistic therapy provided by the present invention uses upconversion luminescent nanoparticles as the luminescence center, which can achieve high-resolution, high-stability and non-contact fluorescence imaging, thereby enabling real-time monitoring of cancer cell sites; using upconversion luminescent nanoparticles as the material carrier, through unique microstructure design, indocyanine green with photothermal therapy and photodynamic therapy functions is combined with manganese dioxide with chemodynamic therapy functions, realizing triple-modal synergistic therapy guided by fluorescence imaging; the multifunctional nanocomposite constructed by the present invention, which integrates the functions of diagnosis and triple-modal therapy, has important application value in the field of cancer diagnosis and treatment.
[0020] 3. The present invention breaks through the limitation of single means in the existing treatment technology and solves the problem of unsatisfactory tumor killing effect and side effect control. Through unique component design and microstructure optimization, the material constructs a new platform with multimodal therapy as the core. This platform supports near-infrared light-driven upconversion fluorescence imaging and its mediated triple-modal synergistic therapy for cancer cells (photodynamic therapy, photothermal therapy and chemodynamic therapy), realizing imaging-guided synergistic therapy, which can significantly enhance the treatment effect and reduce side effects, providing innovative technical ideas and strategies for the field of clinical diagnosis and integrated multimodal treatment.
[0021] 4. The nanocomposite for near-infrared light-driven fluorescence imaging and triple-modal synergistic therapy provided by the present invention and its preparation method have the advantages of high thermal stability, good biocompatibility, etc., and high photothermal conversion efficiency. At the same time, it has the functions of chemodynamic therapy, photothermal therapy and chemodynamic therapy, and realizes the integration of diagnosis and treatment under the guidance of fluorescence imaging.
[0022] 5. The preparation method provided by the present invention is characterized by mild and simple synthesis conditions, high reproducibility, and easy industrialization. Its mild and simple preparation conditions endow the preparation of the nanocomposite system with the advantage of high reproducibility.
[0023] 6. The application of the nanocomposite material provided by the present invention has the functions of fluorescence imaging guided by upconversion luminescent nanoparticles, near-infrared light excitation, multi-enzyme-mediated photothermal therapy and photodynamic therapy, and tumor microenvironment-triggered chemodynamic therapy, realizing the integration of diagnosis and multi-modal therapy. It can achieve accurate diagnosis at the early stage of cancer and be supplemented by efficient treatment means, with potential biomedical application prospects. The multi-enzyme mimicking activity brought by flower-like manganese dioxide is of great significance for improving the treatment effect and reducing side effects in cancer treatment.
[0024] The present invention will be further described in detail below in conjunction with the accompanying drawings and specific embodiments. Description of the Drawings
[0025] Figure 1 It is the transmission electron microscope (TEM) image of the nanocomposite material prepared in Example 1 of the present invention; Figure 2 It is the X-ray photoelectron spectra of UM (UCNP coated with flower-like manganese dioxide), UMN (UCNP coated with amino-functionalized flower-like manganese dioxide), and UMI (UCNP modified with indocyanine green and flower-like manganese dioxide) during the preparation process of the nanocomposite material in Example 2 of the present invention; Figure 3 It is the temperature change of the nanocomposite material in Example 6 of the present invention under 808 nm laser excitation. Specific Embodiments
[0026] The following combines the attached Figures 1-3 and multiple embodiments to clearly and completely describe the technical solutions of the present invention.
[0027] Basic Embodiment
[0028] The near-infrared light-driven fluorescence imaging nanocomposite material provided by the present invention focuses on how to construct a unique microstructure (dual modification, flower-like or honeycomb-like core-shell, etc.) to support synchronous multi-enzyme catalysis and near-infrared laser mediation, and further the laser-mediated triple-modal synergy or multi-modal combination (CDT / PTT / PDT) complementary mechanism to achieve synergistic therapy, significantly improving the tumor treatment effect and reducing side effects.
[0029] The near-infrared light-driven fluorescence imaging nanocomposite provided by this embodiment uses upconversion luminescent nanoparticles UCNP as the core. After depositing flaky manganese dioxide on the surface of the core to form a flower-like or honeycomb-like core-shell structure, the manganese dioxide is then amino-modified, and indocyanine green molecules are further loaded. The obtained upconversion diagnosis and treatment integrated nanocomposite co-modified by indocyanine green molecules and flower-like or honeycomb-like manganese dioxide forms a CDT / PTT / PDT multimodal complementary mechanism under the mediation of multi-enzyme catalysis and near-infrared light.
[0030] The upconversion luminescent nanoparticles UCNP are upconversion luminescent nanoparticles coated with oleic acid. The oleic acid molecules on the surface of UCNP undergo an oxidation-reduction reaction with potassium permanganate to generate flaky manganese dioxide. The manganese dioxide sheets further self-assemble to form a flower-like or honeycomb-like structure. Amines are loaded on the surface of manganese dioxide through the hydrolysis and polycondensation reaction of 3-aminopropyltriethoxysilane in a toluene environment, and the obtained flower-like or honeycomb-like manganese dioxide coated with UCNP is amino-functionalized.
[0031] The amines modified on the surface of manganese dioxide are used to load indocyanine green on the surface of flower-like manganese dioxide through a condensation reaction or a nucleophilic substitution reaction with indocyanine green, and the obtained upconversion diagnosis and treatment integrated nanocomposite functional material co-modified by indocyanine green and flower-like or honeycomb-like manganese dioxide is obtained.
[0032] A preparation method of a near-infrared light-driven fluorescence imaging nanocomposite includes the following steps: S1. Prepare the first dispersion: Prepare a cyclohexane dispersion, an aqueous potassium permanganate solution, and oleic acid containing UCNP respectively. Add oleic acid to the cyclohexane dispersion, ultrasonicate at room temperature and then stir for a period of time to evaporate cyclohexane to obtain an oleic acid dispersion containing UCNP. Then prepare potassium permanganate, dissolve it in a certain amount of deionized water to obtain an aqueous potassium permanganate solution. Mix the aqueous potassium permanganate solution with the oleic acid dispersion containing UCNP, stir at room temperature for a set time, ultrasonically disperse and then centrifuge, and take the solid and disperse it in deionized water to prepare the first dispersion. S2. Prepare the second dispersion: Prepare 3-aminopropyltriethoxysilane, freeze-dry the first dispersion, disperse the obtained powder in a certain amount of toluene solution, ultrasonicate at room temperature and then stir for a period of time. Then add an appropriate amount of 3-aminopropyltriethoxysilane to the above-mentioned stirred mixture, raise the temperature to make it stir and reflux at a set temperature for a set time, centrifuge and take the lower-layer solid and disperse it in deionized water to prepare the second dispersion. S3. Co-modification: Dissolve indocyanine green in a certain amount of deionized water to obtain an indocyanine green aqueous solution. Mix it with the second dispersion liquid in a specified ratio. After stirring in the dark at room temperature for a set time, centrifuge and take the lower-layer solid and disperse it in deionized water, so that indocyanine green undergoes a condensation reaction or nucleophilic substitution reaction with the amino groups on manganese dioxide to form a covalent bond, and then load indocyanine green onto manganese dioxide to obtain an upconversion diagnosis and treatment integrated nano-composite functional material co-modified by indocyanine green and flower-like manganese dioxide, that is, a near-infrared light-driven fluorescence imaging nano-composite material.
[0033] The application of the near-infrared light-driven fluorescence imaging nano-composite material is to use it as a near-infrared light-driven fluorescence imaging contrast agent.
[0034] The application of the near-infrared light-driven fluorescence imaging nano-composite material is to use it as a therapeutic reagent for photothermal therapy, photodynamic therapy, and chemodynamic triple-mode synergistic therapy under near-infrared light excitation, so as to achieve triple-mode synergistic therapy under near-infrared light excitation.
[0035] The application of the near-infrared light-driven fluorescence imaging nano-composite material is to use it as a therapeutic reagent for multi-enzyme catalysis and CDT / PTT / PDT multimodal complementary synergistic therapy mediated by near-infrared light, so as to achieve multi-enzyme catalysis and CDT / PTT / PDT multimodal complementary synergistic therapy mediated by near-infrared light.
[0036] The following combines the attached drawings and multiple specific embodiments to illustrate the detailed technical solutions of the present invention.
[0037] Example 1 The near-infrared light-driven fluorescence imaging nano-composite material, preparation method and its application provided by the embodiment of the present invention are specific embodiments based on the foregoing embodiments. The difference is that the manganese dioxide is constructed in a flower shape, and the preparation method includes the following steps: S1. Add 6 mL of oleic acid solution to the cyclohexane dispersion liquid containing UCNP, stir at 25 °C for 24 hours to obtain an oleic acid dispersion liquid containing UCNP; take 0.031 g of potassium permanganate, add 4 mL of deionized water, ultrasonicate and then stir magnetically until the solution is uniform. Add 8 mL of the oleic acid dispersion liquid containing UCNP to the above solution, ultrasonicate for 10 min and then stir at 25 °C for 5 hours. Use the oleic acid on the surface of UCNP to reduce potassium permanganate to obtain UCNP coated with flower-like manganese dioxide. Then perform ultrasonic dispersion, centrifuge at a speed of 13,000 revolutions per minute to obtain a solid, and disperse it in 10 mL of deionized water to form a first dispersion liquid; S2. Lyophilize 10 mL of the first dispersion to obtain a solid powder. Take 100 mg of the powder and mix it with 30 mL of toluene, stir at 25 °C for 1 hour to form a homogeneous suspension system. Add 500 μL of 3-aminopropyltriethoxysilane to the suspension system, continuously stir at 80 °C for 12 hours to complete the surface amino modification. After ultrasonic dispersion, centrifuge at a speed of 12,000 revolutions per minute, and take the solid and disperse it in 5 mL of deionized water to form a second dispersion; S3. Dissolve 2 mg of indocyanine green in 10 mL of deionized water under light-shielded conditions to obtain an indocyanine green solution; take 5 mL of the second dispersion and mix it with 10 mL of the indocyanine green solution, stir under light-shielded conditions at 25 °C for 24 hours; after ultrasonic dispersion, centrifuge and separate at a rotation speed of 11,000 rpm, take the solid and disperse it in deionized water. The sulfonic acid group in indocyanine green undergoes a condensation reaction or a nucleophilic substitution reaction with the hydroxyl group on the surface of manganese dioxide to form a covalent bond, and then it is loaded onto the surface of manganese dioxide, that is, a diagnostic and therapeutic integrated nanocomposite functional material modified by indocyanine green and manganese dioxide is obtained.
[0038] Figure 1 is the TEM photograph of the near-infrared light-driven fluorescence imaging and multimodal synergistic therapy nanocomposite material prepared in Example 1 of the present invention. It can be observed from Figure 1 that a flower-like manganese dioxide layer loaded with indocyanine green molecules is coated outside the upconversion nanoparticles, indicating that the preparation method can obtain a diagnostic and therapeutic integrated nanocomposite material with good morphology, uniform growth and high experimental repeatability. The average diameter of the nanomaterial is about 55 nm. This morphology helps the nanomaterial to stably exist in the organism and reduces the risk of being rapidly cleared by the organism, which is of great significance for improving its persistence in biological imaging and treatment.
[0039] Example 2 The near-infrared light-driven fluorescence imaging nanocomposite material, preparation method and application provided by the embodiment of the present invention are basically the same as those in Example 1, except that the manganese dioxide is in a honeycomb structure. In this nanocomposite material, oleic acid molecules on the surface of upconversion luminescent nanoparticles (UCNP) undergo an oxidation-reduction reaction with potassium permanganate to generate sheet-like manganese dioxide, and the manganese dioxide lamellae further self-assemble to form a honeycomb structure. After the surface of manganese dioxide is modified with amino groups, a condensation reaction or a nucleophilic substitution reaction occurs with indocyanine green, and indocyanine green is loaded onto the surface of the flower-like manganese dioxide, that is, a diagnostic and therapeutic integrated nanocomposite material modified by honeycomb-like manganese dioxide with multi-enzyme catalytic action and natural molecule indocyanine green is obtained.
[0040] The preparation method of the nanocomposite material includes the following steps: S1. Prepare a cyclohexane dispersion containing UCNP, an aqueous potassium permanganate solution, and oleic acid respectively. Add oleic acid to the cyclohexane dispersion of UCNP, ultrasonicate at room temperature and then stir for a period of time to evaporate cyclohexane, obtaining an oleic acid dispersion of UCNP. Then prepare potassium permanganate, dissolve it in a certain amount of deionized water to obtain an aqueous potassium permanganate solution, mix this aqueous potassium permanganate solution with the oleic acid dispersion of UCNP, stir at room temperature for a set time, ultrasonically disperse and then centrifuge, and disperse the solid in deionized water to prepare the first dispersion. Specifically: Add 6 mL of oleic acid solution to the cyclohexane dispersion containing UCNP, stir at 25 °C for 24 hours to obtain an oleic acid dispersion of UCNP. Take 0.062 g of potassium permanganate, add 4 mL of deionized water, ultrasonicate and then magnetically stir until the solution is uniform. Add 8 mL of the oleic acid dispersion of UCNP to the above solution, ultrasonicate for 10 min and then stir at 25 °C for 5 hours. Use the oleic acid on the surface of UCNP to reduce potassium permanganate to obtain UCNP coated with flower-like manganese dioxide. Then perform ultrasonic dispersion, centrifuge at a speed of 13000 revolutions per minute to obtain a solid, and disperse it in 10 mL of deionized water to form the first dispersion. S2. Prepare 3-aminopropyltriethoxysilane. Freeze-dry the first dispersion, disperse the obtained powder in a certain amount of toluene solution, ultrasonicate at room temperature and then stir for a period of time. Then add an appropriate amount of 3-aminopropyltriethoxysilane to the above stirred mixture, raise the temperature to make it stir and reflux react at a set temperature for a set time, centrifuge and take the lower-layer solid and disperse it in deionized water to prepare the second dispersion. Specifically: Freeze-dry 10 mL of the first dispersion to obtain a solid powder. Take 100 mg of the powder and mix it with 30 mL of toluene, stir at 25 °C for 1 hour to form a uniform suspension system. Add 500 μL of 3-aminopropyltriethoxysilane to the system, continuously stir at 80 °C for 12 hours to complete surface amination modification. Ultrasonically disperse and then centrifuge at a speed of 12000 revolutions per minute, and disperse the solid in 5 mL of deionized water to form the second dispersion. S3. Dissolve indocyanine green in a certain amount of deionized water to obtain an aqueous indocyanine green solution, mix it with the second dispersion in a specified ratio, stir in the dark at room temperature for a set time, centrifuge and take the lower-layer solid and disperse it in deionized water, so that indocyanine green undergoes a condensation reaction or nucleophilic substitution reaction with the amino group on manganese dioxide to form a covalent bond, and then indocyanine green is loaded onto manganese dioxide to obtain a theranostic integrated nanocomposite functional material modified by indocyanine green and honeycomb-like manganese dioxide. Specifically: Dissolve 2 mg of indocyanine green in 10 mL of deionized water under light-shielded conditions. Take 5 mL of the second dispersion and mix it with 10 mL of the indocyanine green solution, and stir it at 25 °C in the dark for 24 hours. After ultrasonic dispersion, centrifuge it at a speed of 11,000 revolutions, take the solid and disperse it in deionized water. The sulfonic acid group in indocyanine green undergoes a condensation reaction or a nucleophilic substitution reaction with the hydroxyl group on the surface of manganese dioxide to form a covalent bond, and then it is loaded onto the surface of manganese dioxide, that is, a diagnostic and therapeutic integrated nano-composite functional material modified by indocyanine green and manganese dioxide is obtained.
[0041] Figure 2 It is the near-infrared light-driven fluorescence imaging nano-composite material prepared in this example. During the preparation process, the X-ray photoelectron spectra of UM (UCNP coated with honeycomb-like manganese dioxide), UMN (UCNP coated with amino-functionalized flower-like manganese dioxide), and UMI (UCNP modified with indocyanine green and honeycomb-like manganese dioxide) are shown. Five obvious peaks are shown at 685, 642, 399, 285, and 159 eV, corresponding to F, Mn, N, C, and S elements respectively. The F element comes from the UCNP nanoparticles, while the Mn, N, C, and S elements come from the manganese dioxide coating layer, the amino-functionalized modification reagent, and indocyanine green. The accurate attribution of these peak positions can illustrate the successful preparation of the composite material provided by the present invention. This benefits from the simple, mild, and easy-to-control conditions of the preparation method provided by the present invention, so that the preparation of the nano-composite material has high repeatability. After testing, the honeycomb-like manganese dioxide layer provided in this example and the flower-like manganese dioxide layer in Example 1 have highly similar CDT effects and catalytic characteristics, so they will not be elaborated here.
[0042] Example 3 The near-infrared light-driven fluorescence imaging nano-composite material, preparation method, and application provided by the embodiments of the present invention are basically the same as those in Examples 1 and 2. The difference is that the oleic acid molecules on the surface of the upconversion luminescent nanoparticles (UCNP) undergo an oxidation-reduction reaction with potassium permanganate to generate sheet-like manganese dioxide, and then the manganese dioxide sheets further self-assemble to form a stacked flower-like structure. After the surface of the manganese dioxide is modified with amino groups, a condensation reaction or a nucleophilic substitution reaction occurs with indocyanine green, and indocyanine green is loaded onto the surface of the stacked flower-like manganese dioxide, that is, a diagnostic and therapeutic integrated nano-composite material modified by stacked flower-like manganese dioxide with multi-enzyme catalytic action and the natural molecule indocyanine green is obtained. Among them, the amino-functionalized modification reagent is 3-aminopropyltrimethoxysilane.
[0043] The preparation method of the above-mentioned near-infrared light-driven fluorescence imaging nano-composite material includes the following steps: S1. Prepare a cyclohexane dispersion containing UCNP, an aqueous potassium permanganate solution, and oleic acid respectively. Add oleic acid to the cyclohexane dispersion containing UCNP, ultrasonicate at room temperature and then stir for a period of time to evaporate cyclohexane, obtaining an oleic acid dispersion of UCNP. Then prepare potassium permanganate, dissolve it in a certain amount of deionized water to obtain an aqueous potassium permanganate solution, mix this aqueous potassium permanganate solution with the oleic acid dispersion of UCNP, stir at room temperature for a set time, ultrasonically disperse and then centrifuge, take the solid and disperse it in deionized water to prepare the first dispersion. Specifically: Add 6 mL of oleic acid solution to the cyclohexane dispersion containing UCNP, stir at 25 °C for 24 hours to obtain an oleic acid dispersion of UCNP. Take 0.062 g of potassium permanganate, add 4 mL of deionized water, ultrasonicate and then magnetically stir until the solution is uniform. Add 8 mL of the oleic acid dispersion of UCNP to the above solution, ultrasonicate for 10 min and then stir at 25 °C for 5 hours. Use the oleic acid on the surface of UCNP to reduce potassium permanganate to obtain UCNP coated with flower-like manganese dioxide. Then perform ultrasonic dispersion, centrifuge at a speed of 13000 revolutions per minute to obtain a solid, and disperse it in 10 mL of deionized water to form the first dispersion. S2. Prepare 3-aminopropyltrimethoxysilane. Freeze-dry the first dispersion, disperse the obtained powder in a certain amount of toluene solution, ultrasonicate at room temperature and then stir for a period of time. Then add an appropriate amount of 3-aminopropyltriethoxysilane to the above stirred mixture, raise the temperature and stir and reflux at a set temperature for a set time, centrifuge and take the lower-layer solid and disperse it in deionized water to prepare the second dispersion. Specifically: Freeze-dry 10 mL of the first dispersion to obtain a solid powder. Take 100 mg of the powder and mix it with 30 mL of toluene, stir at 25 °C for 1 hour to form a uniform suspension system. Add 500 μL of 3-aminopropyltrimethoxysilane to the system, continuously stir at 110 °C for 12 hours to complete the surface amino modification. After ultrasonic dispersion, centrifuge at a speed of 12000 revolutions per minute, take the solid and disperse it in 5 mL of deionized water to form the second dispersion. S3. Dissolve indocyanine green in a certain amount of deionized water to obtain an aqueous indocyanine green solution, mix it with the second dispersion in a specified ratio, stir in the dark at room temperature until the set time, centrifuge and take the lower-layer solid and disperse it in deionized water to enable indocyanine green to undergo a condensation reaction or nucleophilic substitution reaction with the amino group on manganese dioxide to form a covalent bond, and then indocyanine green is loaded onto manganese dioxide to obtain an upconversion diagnosis and treatment integrated nano-composite functional material modified with indocyanine green and layered flower-like manganese dioxide. Specifically: Dissolve 2 mg of indocyanine green in 10 mL of deionized water under light - avoiding conditions. Take 5 mL of the second dispersion and mix it with 10 mL of the indocyanine green solution, and stir magnetically for 24 hours at 25 °C under light - avoiding conditions. After ultrasonic dispersion, centrifuge at a speed of 11,000 rpm, take the solid and disperse it in deionized water. The sulfonic acid group in indocyanine green undergoes a condensation reaction or a nucleophilic substitution reaction with the hydroxyl group on the surface of manganese dioxide to form a covalent bond, and then it is loaded onto the surface of manganese dioxide, that is, a diagnosis - treatment integrated nano - composite functional material modified by indocyanine green and manganese dioxide is obtained.
[0044] Example 4 In this example, the near - infrared light - driven fluorescence imaging nano - composite materials prepared in the above Examples 1 - 3 are specifically applied. Specifically, the near - infrared light - driven fluorescence imaging and triple - modality synergistic therapy nano - composite materials prepared in one of Examples 1 - 3 are used. The up - conversion luminescent nanoparticles therein are used as the luminescence center of the material for fluorescence imaging with both high resolution and high stability, and can realize real - time monitoring of cancer cell sites; the up - conversion luminescent nanoparticles therein are used as the material carrier. Under the premise of unique component and microstructure design, they are combined with indocyanine green with photothermal therapy and photodynamic therapy functions and manganese dioxide with chemodynamic therapy functions to achieve triple - modality therapy guided by fluorescence imaging; the multifunctional nano - composite material integrating diagnosis and multi - modality therapy functions constructed in this example has good application potential and provides a new strategy for the field of clinical diagnosis and multi - mode therapy integration.
[0045] Example 5 In this example, the composite material prepared in Example 1 is specifically used as a material for preparing photothermal reagents, thermokinetic therapy reagents, and chemokinetic therapy reagents for near - infrared light - excited photothermal therapy. This composite material simultaneously supports laser - mediated imaging and triple - modality therapy and can be used as a reagent or medicine, which is widely used in in vitro photothermal imaging diagnosis and treatment activities.
[0046] This example is a method for using the prepared near - infrared light - driven fluorescence imaging nano - composite material for in vitro photothermal therapy under 808 nm laser excitation, which includes the following steps: (1) Prepare the near - infrared light - driven fluorescence imaging nano - composite material in Example 1, a quartz cuvette, and a continuously - excited 808 nm laser; (2) Take the nano - composite material prepared in Example 1 and prepare solutions with concentrations of 0, 50 μg / mL, 100 μg / mL, 200 μg / mL, and 400 μg / mL with deionized water, and disperse them evenly by ultrasonic treatment to form third dispersions with different concentrations; (3) Respectively take 1.5 mL of the fourth dispersions with different concentrations and add them to the quartz cuvette, and fix it on an iron stand. Use 1 W / cm 2The third dispersion liquid was continuously irradiated with an 808 nm laser, and the real-time temperature of the third dispersion liquid at different times was recorded using a temperature sensor.
[0047] Figure 3 It is a schematic diagram of the test results of the near-infrared light-driven fluorescence imaging nanocomposite obtained in Example 1 of the present invention for in vitro photothermal experiments. It can be seen from the figure that under the excitation of an 808 nm laser, as the concentration of the third dispersion liquid increases, the temperature also gradually rises; at the same time, the temperature of the dispersion liquid also increases with the delay of the irradiation time. After 8 minutes, the temperature of the solution has tended to be stable, reaching as high as 43 °C. Existing studies have shown that under the condition of 42 °C, culturing with tumor cells for 1 hour can achieve excellent photothermal therapy and kill cancer cells. This characterization indicates that the nanocomposite provided by the present invention has great application prospects in improving the photothermal therapy effect of tumor cells and reducing side effects.
[0048] The near-infrared light-driven fluorescence imaging nanocomposite, preparation method and application provided in the above embodiments of the present invention mainly involve upconversion luminescent nanoparticles coated with oleic acid. The oleic acid molecules on the surface undergo an oxidation-reduction reaction with potassium permanganate to form flaky manganese dioxide. The flaky manganese dioxide further accumulates to form a flower-shaped core-shell structure. The hydroxyl groups on the surface of manganese dioxide form silicon-oxygen-manganese covalent bonds with silanol, the hydrolysis product of the silane coupling agent, and further fix amino groups on the surface of manganese dioxide. Indocyanine green undergoes a condensation or nucleophilic substitution reaction with the surface amino groups and is loaded on the surface of flower-shaped or honeycomb-shaped manganese dioxide to obtain a near-infrared light-driven fluorescence imaging and triple-modal synergistic therapy nanocomposite. The preparation method of the nanocomposite provided by the present invention is simple, mild and highly reproducible. The obtained nanocomposite has a uniform particle size and good biocompatibility, and can be applied to biomedical fields such as triple-modal synergistic therapy including fluorescence imaging-mediated photodynamic therapy, photothermal therapy and chemodynamic therapy of cancer cells.
[0049] The present invention provides an application of an infrared light-driven fluorescence imaging nanocomposite. The fluorescence imaging-mediated triple-modal synergistic therapy for cancer cells is a cancer treatment method that combines fluorescence imaging, photodynamic therapy (PDT), photothermal therapy (PTT), and chemodynamic therapy (CDT). This diagnostic and treatment method utilizes the complementary advantages of different treatment modalities to improve the treatment effect and reduce side effects. The fluorescence imaging-mediated triple-modal synergistic therapy for cancer cells achieves multiple strikes on cancer cells by combining three treatment modalities: PDT, PTT, and CDT. Specifically, fluorescence imaging is used to localize and monitor tumor tissues. PDT and CDT generate reactive oxygen species (ROS) to destroy cancer cells, while PTT generates heat to kill cancer cells. This synergistic effect can enhance the treatment effect, reduce the limitations of a single treatment modality, and reduce damage to normal tissues. It combines the advantages of different treatment modalities and provides a new technical idea for improving the cancer treatment effect and reducing side effects.
[0050] As described above, these are merely preferred embodiments of the present invention and do not impose any form of limitation on the present invention. Any person skilled in the art can make many possible changes or equivalent changes to the technical solution of the present invention within the scope of the technical solution of the present invention by using the methods and technical contents disclosed above. Therefore, any equivalent modification made based on the structure, construction, and principle of the present invention without departing from the content of the present technical invention solution should be covered within the protection scope of the present invention.
Claims
1. A near-infrared light-driven fluorescence imaging nanocomposite, characterized in that, It is a core-shell structure with the above-mentioned upconversion luminescent nanoparticles UCNP as the core, and flaky manganese dioxide is deposited on the surface of the core to form a flower-like or honeycomb-like core-shell structure. Then, the manganese dioxide is amino-modified, and indocyanine green molecules are further loaded. The obtained upconversion diagnosis and treatment integrated nanocomposite material modified by indocyanine green molecules and flower-like or honeycomb-like manganese dioxide forms a CDT / PTT / PDT multimodal complementary mechanism under the mediation of multi-enzyme catalysis and near-infrared light.
2. The near-infrared light-driven fluorescence imaging nanocomposite according to claim 1, wherein The above-mentioned upconversion luminescent nanoparticles UCNP are upconversion luminescent nanoparticles coated with oleic acid. The oleic acid molecules on the surface of UCNP undergo an oxidation-reduction reaction with potassium permanganate to generate flaky manganese dioxide. The manganese dioxide lamellae further self-assemble to form a flower-like or honeycomb-like structure. Amino groups are loaded on the surface of manganese dioxide through the hydrolysis and polycondensation reaction of 3-aminopropyltriethoxysilane in a toluene environment, and the obtained UCNP is coated with amino-functionalized manganese dioxide.
3. The near-infrared light-driven fluorescence imaging nanocomposite according to claim 1, wherein The amino groups modified on the surface of manganese dioxide load indocyanine green onto the surface of flower-like or honeycomb-like manganese dioxide through a condensation reaction or a nucleophilic substitution reaction with indocyanine green, and a upconversion diagnosis and treatment integrated nanocomposite functional material modified by indocyanine green and manganese dioxide is obtained.
4. A method for preparing a near-infrared light-driven fluorescence imaging nanocomposite, characterized in that, It includes the following steps: S1. Prepare the first dispersion: Prepare a cyclohexane dispersion, an aqueous potassium permanganate solution, and oleic acid containing UCNP respectively; add oleic acid to the cyclohexane dispersion, ultrasonically stir at room temperature for a period of time, and then evaporate cyclohexane to obtain an oleic acid dispersion containing UCNP; then prepare potassium permanganate, dissolve it in a certain amount of deionized water to obtain an aqueous potassium permanganate solution, mix the aqueous potassium permanganate solution with the oleic acid dispersion containing UCNP, stir at room temperature for a set time, ultrasonically disperse, and then centrifuge. Take the solid and disperse it in deionized water to prepare the first dispersion. S2. Prepare the second dispersion: Prepare 3-aminopropyltriethoxysilane, freeze-dry the first dispersion, and disperse the obtained powder in a certain amount of toluene solution, ultrasonically stir at room temperature for a period of time; then add an appropriate amount of 3-aminopropyltriethoxysilane to the above-mentioned stirred mixture, raise the temperature to make it stir and reflux at a set temperature for a set time, centrifuge, and take the lower-layer solid and disperse it in deionized water to prepare the second dispersion. S3. Co-modification: Dissolve indocyanine green in a certain amount of deionized water to obtain an indocyanine green aqueous solution, mix it with the second dispersion in a specified ratio, stir in the dark at room temperature for a set time, centrifuge, and take the lower-layer solid and disperse it in deionized water to make indocyanine green undergo a condensation reaction or a nucleophilic substitution reaction with the amino groups on manganese dioxide to form a covalent bond, and further load indocyanine green onto manganese dioxide to obtain a upconversion diagnosis and treatment integrated nanocomposite functional material modified by indocyanine green and flower-like or honeycomb-like manganese dioxide, that is, a near-infrared light-driven fluorescence imaging nanocomposite material.
5. The preparation method according to claim 4, characterized in that, The specific steps for preparing the first dispersion in step S1 are as follows: S1-1 Add 6 mL of oleic acid solution to the cyclohexane dispersion containing UCNP, stir at 25 °C for 24 hours to obtain an oleic acid dispersion containing UCNP. S1-1 Take 0.02 - 0.04 g of potassium permanganate, add 4 mL of deionized water, ultrasonicate and then stir magnetically until the solution is homogeneous. S1-2 Add 8 mL of oleic acid dispersion containing UCNP to the above solution, ultrasonicate for 10 min and then stir at 25 °C for 5 - 8 hours. Reduce potassium permanganate with the oleic acid on the surface of UCNP to obtain UCNP coated with flower-like manganese dioxide. S1-3 Then perform ultrasonic dispersion, centrifuge at a speed of 13,000 revolutions per minute to obtain a solid, and disperse it in 10 mL of deionized water to obtain the first dispersion.
6. The preparation method according to claim 4, characterized in that, The specific steps for preparing the second dispersion in step S2 are as follows: S2-1 Take 10 mL of the first dispersion and freeze-dry it to obtain a solid powder. Take 50 - 100 mg of the powder and mix it with 20 - 30 mL of toluene, and stir at 25 °C for 1 hour to form a homogeneous suspension system. S2-2 Add 400 - 600 μL of 3-aminopropyltriethoxysilane to the suspension system, continuously stir at 75 - 85 °C for 12 hours to complete surface amination modification. After ultrasonic dispersion, centrifuge at a speed of 11,000 revolutions per minute, and take the solid and disperse it in 5 mL of deionized water to obtain the second dispersion.
7. The preparation method according to claim 4, characterized in that, The specific steps for obtaining the final material in step S3 are as follows: S3-1 Dissolve 1 - 3 mg of indocyanine green in 10 mL of deionized water under light-shielded conditions to obtain an indocyanine green solution. S3-2 Take 5 mL of the second dispersion and mix it with 10 mL of the indocyanine green solution, and stir at 25 °C in the dark for 24 hours to obtain a mixed solution. After ultrasonic dispersion of the mixed solution, centrifuge and separate it at a speed of 11,000 rpm. Take the solid and disperse it in deionized water, so that the sulfonic acid group in indocyanine green undergoes a condensation reaction or nucleophilic substitution reaction with the hydroxyl group on the surface of manganese dioxide to form a covalent bond, and then load it onto the surface of manganese dioxide, that is, a diagnosis and treatment integrated nano-composite functional material co-modified by indocyanine green and manganese dioxide is obtained.
8. Use of the near-infrared light-driven fluorescence imaging nanocomposite according to any one of claims 1 to 3, characterized in that, Use it as a near-infrared light-driven fluorescence imaging contrast agent.
9. Use of the near-infrared light-driven fluorescence imaging nanocomposite according to any one of claims 1 to 3, characterized in that Use it as a photothermal therapy reagent, a photosensitizer for photodynamic therapy and an enhancing reagent, and a treatment reagent for triple-mode synergistic therapy of chemodynamic therapy under near-infrared light excitation.
10. Use of the near-infrared light-driven fluorescence imaging nanocomposite according to any one of claims 1 to 3, characterized in that, Use it as a treatment reagent for multi-enzyme catalysis and multi-modal complementary synergistic therapy of CDT / PTT / PDT under near-infrared light mediation.