Tumor microenvironment activation sound-sensitive agent, preparation method and application
By designing a Cu-IR783 sound-sensitizer activated in the acid tumor microenvironment, the biocompatibility and targeting of existing sound-sensitizers is solved, efficient and visual anti-tumor treatment is achieved, and side effects are minimized.
Patent Information
- Application Number
- CN202510083627.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-20
- Publication Date
- 2025-04-15
AI Technical Summary
During the development of existing sound-sensitizers, biocompatibility and biodegradability have been ignored, resulting in potential safety problems; at the same time, the sound-sensitizers administered intravenously lack active targeting, which may cause irreversible damage to normal tissues, and it is difficult to monitor the accumulation time and therapeutic effect of sound-sensitizers in tumor tissues.
A tumor microenvironment activated sound sensitizer was designed to form a Cu-IR783 sound sensitizer that forms nanoparticles through coordination assembly of copper ions and IR783 near-infrared dyes. Under normal physiological conditions, the acoustic dynamic activity and fluorescence imaging ability of the acoustic sensitive agent are passivated and reactivated only in the acidic tumor microenvironment to achieve in-situ visual acoustic dynamic therapy.
Cu-IR783 sound sensitizer activated by TME can significantly improve ROS levels, enhance anti-tumor performance, and monitor treatment effects through near-infrared imaging to achieve intelligent, visual and in-situ anti-tumor treatment, and minimize treatment side effects.
Smart Images
Figure CN119818673A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the field of biomedical application technology, and in particular to a tumor microenvironment activating sonosensitizer, a preparation method and uses thereof. Background Art
[0002] In the process of tumor progression, reactive oxygen species is a double-edged sword. Low levels of ROS are involved in the growth process of tumors, while high levels of ROS can trigger oxidative stress responses in tumor cells, leading to cell apoptosis. In recent years, nanotechnology that uses reactive oxygen species (ROS) to fight cancer has received widespread attention. Some new tumor treatment methods that use exogenous stimuli or endogenous chemical reactions to generate ROS have been widely studied, such as photodynamic therapy, sonodynamic therapy, and chemodynamic therapy. Among them, sonodynamic therapy (SDT) has the advantages of high tissue penetration depth and treatment is not limited by depth. In addition, the acoustic cavitation effect can promote the penetration of nanomedicines into the tumor through blood vessels, thereby enhancing the therapeutic effect.
[0003] Through the analysis of the SDT mechanism, it can be seen that the performance of sonosensitizers and the influence of tumor microenvironment (TME) play a decisive role in the efficacy of SDT. In order to improve the performance of SDT, people have explored many strategies, such as developing sonosensitizers with narrow band gaps, introducing nanozymes with multi-enzyme activities to break the TME restrictions, and combining with other treatment methods. However, the development of SDT still faces many challenges. First, the current development of sonosensitizers usually only focuses on improving the efficiency of ROS generation, while ignoring the biocompatibility and biodegradability of sonosensitizers, which may lead to potential safety issues. Secondly, the sonosensitizers administered intravenously lack active targeting, and will also accumulate in normal tissues and induce sonodynamic effects, which may cause irreversible damage to tissues or organs. Third, it is difficult to intuitively monitor the precise accumulation time and therapeutic effect of sonosensitizers in tumor tissues. Although it has been reported that small molecule dyes are combined with sonosensitizers for imaging-mediated cancer treatment, the inevitable accumulation in normal tissues makes it difficult to visualize and monitor the therapeutic effect. To address the above issues, it is a major challenge to design a smart sonosensitizer with an “on / off” function similar to the principle of a prodrug, that is, to passivate the sonodynamic activity of the sonosensitizer and quench the imaging ability of the fluorescent probe under normal physiological conditions, and only reactivate it in the tumor microenvironment to achieve in situ visualized sonodynamic therapy.
[0004] In view of this, the present invention provides a tumor microenvironment activating sonosensitizer, a preparation method and use thereof. Summary of the invention
[0005] The purpose of the present invention is to provide a tumor microenvironment activating sonosensitizer, a preparation method and use thereof in view of the deficiencies in the prior art.
[0006] In order to solve the above technical problems, the following technical solutions are adopted: A tumor microenvironment activated sonosensitizer comprises copper ions and IR783 near-infrared dye. The copper ions and IR783 near-infrared dye are coordinated and assembled to obtain a nanoparticle Cu-IR783 sonosensitizer.
[0007] The present invention proposes another technical solution: a method for preparing a tumor microenvironment activated sonosensitizer, comprising the following steps: S1. Add the copper ions and IR783 near-infrared dye to phosphate buffer, stir at room temperature for 20-30 hours to obtain a mixed solution, wherein the mass ratio of the copper ions, IR783 near-infrared dye and phosphate buffer is 1: (3.5-4.5): (95-105).
[0008] S2. The mixed solution in step S1 is centrifuged and washed with deionized water for 2-5 times to obtain pure nanoparticle Cu-IR783 sonosensitizer.
[0009] The present invention proposes another technical solution: a use of a tumor microenvironment activated sonosensitizer, taking the Cu-IR783 sonosensitizer prepared above, testing the sonodynamic activity of the Cu-IR783 sonosensitizer, and using DPBF as 1 O2 probe, detects the Cu-IR783 sonosensitizer produced under ultrasonic irradiation 1 The levels of O2 were compared with the same concentration of IR783 near-infrared dye to verify the passivation of its sonodynamic activity.
[0010] The present invention proposes another technical solution: a use of a tumor microenvironment activated sonosensitizer, taking the Cu-IR783 sonosensitizer prepared as above, testing the glutathione consumption of the Cu-IR783 sonosensitizer, and evaluating the ability of the IR783 near-infrared dye and the Cu-IR783 sonosensitizer to consume GSH under ultrasonic irradiation by using DTNB as a GSH probe.
[0011] The present invention proposes another technical solution: a use of a tumor microenvironment activated sonosensitizer, taking the Cu-IR783 sonosensitizer prepared as above, and testing the near-infrared imaging capability of the Cu-IR783 sonosensitizer.
[0012] The fluorescence intensity of the IR783 near-infrared dye and the Cu-IR783 sonosensitizer at the optimal emission peak was detected using a fluorescence spectrometer to evaluate their near-infrared fluorescence quenching.
[0013] The present invention proposes another technical solution: a use of a tumor microenvironment activated sonosensitizer, taking the Cu-IR783 sonosensitizer prepared as above, and testing the near-infrared imaging capability of the Cu-IR783 sonosensitizer.
[0014] The near-infrared fluorescence changes of the IR783 near-infrared dye and the Cu-IR783 sonosensitizer are detected by an in vivo imaging monitoring system to evaluate their fluorescence imaging capabilities.
[0015] The present invention proposes another technical solution: a use of a tumor microenvironment activated sonosensitizer, taking the Cu-IR783 sonosensitizer prepared as above, and testing the acidic TME response decomposition behavior of the Cu-IR783 sonosensitizer.
[0016] The Cu-IR783 sonosensitizer prepared by the present invention is placed in different pH buffer solutions of pH 6.0, pH 6.5 and pH 7.4 for dialysis for 7 days; the Cu-IR783 sonosensitizer after dissociation every day is measured, and the absorption spectrum and fluorescence spectrum of the Cu-IR783 sonosensitizer are measured to evaluate the dissociation behavior of the Cu-IR783 sonosensitizer.
[0017] The present invention proposes another technical solution: a use of a tumor microenvironment activated sonosensitizer, taking the Cu-IR783 sonosensitizer prepared as above, and testing the acidic TME response decomposition behavior of the Cu-IR783 sonosensitizer.
[0018] The Cu-IR783 sonosensitizer prepared by the present invention was placed in different pH buffer solutions of pH 6.0, pH 6.5 and pH 7.4 for dialysis for 7 days; 1 The O2 probe and the GSH probe are used to detect the sonodynamic activity and GSH consumption ability of the dissociated Cu-IR783 sonosensitizer.
[0019] The present invention proposes another technical solution: a use of a tumor microenvironment activated sonosensitizer, taking the Cu-IR783 sonosensitizer prepared as above, and testing the acidic TME response decomposition behavior of the Cu-IR783 sonosensitizer.
[0020] The Cu-IR783 sonosensitizer prepared by the present invention was placed in different pH buffer solutions of pH 6.0, pH 6.5 and pH 7.4 for dialysis for 7 days; TMB was used as a probe to detect the ability of the Cu-IR783 sonosensitizer to generate hydroxyl radicals in a buffer solution containing H2O2.
[0021] The present invention proposes another technical solution: a use of a tumor microenvironment activated sonosensitizer, taking the Cu-IR783 sonosensitizer prepared as above, testing the in vitro anti-tumor performance of the Cu-IR783 sonosensitizer, and detecting the survival rate of brain glioma cells treated with the Cu-IR783 sonosensitizer and ultrasonic irradiation by the MTT method.
[0022] The brain glioma cells were seeded in a 96-well plate at 5000 cells / well. After incubation overnight, the old culture medium was replaced with fresh culture medium containing different concentrations of the Cu-IR783 sonosensitizer, and the culture was continued for 24 or 48 hours. At the 4th hour, the cells were ultrasonically treated. Then, the in vitro anti-tumor efficacy of the Cu-IR783 sonosensitizer was detected according to the standard MTT protocol.
[0023] The above technical solution has the following beneficial effects: The present invention relates to the field of biomedical applications, and in particular to the preparation of a tumor microenvironment activated sonosensitizer, and discloses the application of the sonosensitizer as an intelligent sonosensitizer in in situ visualized anti-tumor treatment. 2+ ) and the coordination assembly ability of IR783 near-infrared dye, a Cu-IR783 smart sonosensitizer with passivated sonodynamic activity and near-infrared imaging capabilities was designed and constructed. Under normal physiological conditions, the near-infrared imaging and sonodynamic activity of the Cu-IR783 sonosensitizer nanoparticles are passivated, but because the coordination bond between the IR783 near-infrared dye and the copper ion is broken in the acidic TME (acidic tumor microenvironment), the near-infrared imaging and SDT effects are selectively activated and a Fenton-like reaction is triggered. Through TME activation, the ROS level caused by the Cu-IR783 sonosensitizer is greatly increased and copper death occurs, reversing the immunosuppressive tumor microenvironment, and triggering immunogenic cell death to promote systemic immune response, thereby achieving excellent anti-tumor performance.
[0024] The present invention integrates sonodynamic therapy, chemodynamic therapy and copper death in a controllable manner to achieve intelligent, visualized and in situ anti-tumor treatment strategies and minimize treatment side effects. BRIEF DESCRIPTION OF THE DRAWINGS
[0025] The present invention will be further described below in conjunction with the accompanying drawings: Figure 1 TEM image of the Cu-IR783 sonosensitizer of the present invention.
[0026] Figure 2 XPS spectrum analysis diagram of the Cu-IR783 sonosensitizer of the present invention.
[0027] Figure 3 The results of the sonodynamic activity (a) and glutathione consumption capacity (b) of the IR783 near-infrared dye and Cu-IR783 smart sonosensitizer of the present invention are shown in FIG.
[0028] Figure 4 The near-infrared imaging capability passivation diagram of the IR783 near-infrared dye and Cu-IR783 smart sonosensitizer of the present invention.
[0029] Figure 5TME response decomposition behavior diagram of the Cu-IR783 intelligent sonosensitizer of the present invention.
[0030] Figure 6 The result diagram of the sonodynamic activity (a) and GSH consumption capacity (b) of the Cu-IR783 intelligent sonosensitizer of the present invention under different pH conditions (6.0, 7.4) and ultrasonic irradiation.
[0031] Figure 7 The results of the TME response decomposition behavior (a) and cytotoxicity experiment (b) of the Cu-IR783 intelligent sonosensitizer of the present invention at the cellular level are shown in the figure.
[0032] Figure 8 Figure 2 is a graph showing the growth of brain tumor cells in mice during treatment with the present invention.
[0033] Fig. 9 The present invention shows the volume changes of primary tumor (a) and distal tumor (b) in mice after different treatments in the bilateral tumor model. DETAILED DESCRIPTION
[0034] In order to make the purpose, technical scheme and advantages of the present invention clearer, the present invention is further described in detail below through the accompanying drawings and embodiments. However, it should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the scope of the present invention. In addition, in the following description, the description of known structures and technologies is omitted to avoid unnecessary confusion of the concept of the present invention.
[0035] See also Figure 1 and Figure 2 A tumor microenvironment activated sonosensitizer includes copper ions and IR783 near-infrared dye. The copper ions and IR783 near-infrared dye are coordinated and assembled to obtain a nanoparticle Cu-IR783 sonosensitizer.
[0036] Based on the above-mentioned Cu-IR783 sonosensitizer, a method for preparing a tumor microenvironment activated sonosensitizer is provided, comprising the following steps: S1. Add the copper ions and IR783 near-infrared dye to phosphate buffer, stir at room temperature for 20-30 hours to obtain a mixed solution, wherein the mass ratio of the copper ions, IR783 near-infrared dye and phosphate buffer is 1: (3.5-4.5): (95-105).
[0037] Specifically, in this embodiment, 0.1 g of IR783 near-infrared dye and 0.4 g of copper chloride (CuCl2) were added to 10 mL of phosphate buffer, wherein the pH value of the phosphate buffer was controlled to be 7.4, and stirred at room temperature for 24 h.
[0038] S2. The mixed solution in step S1 is centrifuged and washed with deionized water for 2-5 times to obtain pure nanoparticle Cu-IR783 sonosensitizer. Specifically, in this embodiment, the number of washing times with deionized water is 3 times.
[0039] In this embodiment, Depend on Figure 1 It can be seen that the Cu-IR783 intelligent sonosensitizer prepared by the present invention is a well-dispersed spherical nanoparticle with a size of about 50-60 nm.
[0040] Depend on Figure 2 It can be seen that the XPS spectrum shows the presence of Cu, C, S, O and N elements, proving the successful preparation of the Cu-IR783 intelligent sonosensitizer.
[0041] Based on the above-mentioned Cu-IR783 sonosensitizer, a use of a tumor microenvironment activation sonosensitizer is provided. The Cu-IR783 sonosensitizer prepared above is taken to test the sonodynamic activity of the Cu-IR783 sonosensitizer. By using DPBF as 1 O2 probe, detects the Cu-IR783 sonosensitizer produced under ultrasonic irradiation 1 The levels of O2 were compared with the same concentration of IR783 near-infrared dye to verify the passivation of its sonodynamic activity.
[0042] in, 1 The O2 probe is singlet oxygen ( 1 O2) probe.
[0043] Based on the above-mentioned Cu-IR783 sonosensitizer, a use of a tumor microenvironment-activated sonosensitizer is provided. The Cu-IR783 sonosensitizer prepared above is taken to test the glutathione consumption of the Cu-IR783 sonosensitizer. By using DTNB as a GSH probe, the ability of the IR783 near-infrared dye and the Cu-IR783 sonosensitizer to consume GSH under ultrasound irradiation is evaluated.
[0044] Among them, DTNB is 5,5'-dithiobis(2-nitrobenzoic acid).
[0045] Depend on Figure 3 It can be seen that compared with the IR783 near-infrared dye with high sonodynamic activity, the Cu-IR783 intelligent sonosensitizer prepared by the present invention has lost its sonodynamic activity and cannot generate 1 O2 and consumption of GSH. This indicates that the sonodynamic activity of the IR783 near-infrared dye is passivated after assembly with Cu ions.
[0046] Based on the above-mentioned Cu-IR783 sonosensitizer, a use of a tumor microenvironment activated sonosensitizer is provided, the Cu-IR783 sonosensitizer prepared above is taken, and the near-infrared imaging ability of the Cu-IR783 sonosensitizer is tested, The fluorescence intensity of the IR783 near-infrared dye and the Cu-IR783 sonosensitizer at the optimal emission peak was detected using a fluorescence spectrometer to evaluate their near-infrared fluorescence quenching.
[0047] Based on the above-mentioned Cu-IR783 sonosensitizer, a use of a tumor microenvironment activated sonosensitizer is provided, the Cu-IR783 sonosensitizer prepared above is taken, and the near-infrared imaging ability of the Cu-IR783 sonosensitizer is tested, The near-infrared fluorescence changes of the IR783 near-infrared dye and the Cu-IR783 sonosensitizer are detected by an in vivo imaging monitoring system to evaluate their fluorescence imaging capabilities.
[0048] Depend on Figure 4 It can be seen that after assembly with Cu ions, the near-infrared fluorescence of IR783 is quenched, indicating that the near-infrared imaging ability of IR783 disappears.
[0049] Based on the above-mentioned Cu-IR783 sonosensitizer, a use of a tumor microenvironment-activated sonosensitizer is provided. The Cu-IR783 sonosensitizer prepared above is taken, and the acidic TME response decomposition behavior of the Cu-IR783 sonosensitizer is tested. The Cu-IR783 sonosensitizer prepared by the present invention is placed in different pH buffer solutions of pH 6.0, pH 6.5 and pH 7.4 for dialysis for 7 days; the Cu-IR783 sonosensitizer after dissociation every day is taken for measurement, and the absorption spectrum and fluorescence spectrum of the Cu-IR783 sonosensitizer are measured to evaluate the dissociation behavior of the Cu-IR783 sonosensitizer.
[0050] The Cu-IR783 sonosensitizer after dissociation is taken for measurement every day. Different time points of each day can be selected as needed, or the same time point of each day can be selected.
[0051] See also Figure 5 , Absorption spectra of Cu-IR783 intelligent sonosensitizer dissociated under different pH conditions ( Figure 5 a) and fluorescence spectra ( Figure 5 b); Near infrared images of Cu-IR783 intelligent sonosensitizer after incubation for different time periods under different pH conditions (6.0, 7.4) ( Figure 5 c). Figure 5It can be seen that the characteristic peak intensity in the absorption spectrum of the Cu-IR783 smart sonosensitizer increases with the decrease of pH value, and the near-infrared fluorescence of IR783 is restored. At the same time, the Cu-IR783 smart sonosensitizer will not decompose in a neutral environment, indicating that the Cu-IR783 smart sonosensitizer only releases IR783 under acidic conditions and the near-infrared imaging ability is restored.
[0052] Based on the above-mentioned Cu-IR783 sonosensitizer, a use of a tumor microenvironment activation sonosensitizer is provided, the Cu-IR783 sonosensitizer prepared above is taken, and the acidic TME response decomposition behavior of the Cu-IR783 sonosensitizer is tested, and the Cu-IR783 sonosensitizer prepared by the present invention is placed in different pH buffer solutions of pH 6.0, pH 6.5 and pH 7.4 for 7 days; by using 1 O2 probe and GSH probe, detecting the sonodynamic activity and GSH consumption ability of the dissociated Cu-IR783 sonosensitizer; Based on the above-mentioned Cu-IR783 sonosensitizer, a use of a tumor microenvironment-activated sonosensitizer is provided. The Cu-IR783 sonosensitizer prepared above is taken, and the acidic TME response decomposition behavior of the Cu-IR783 sonosensitizer is tested. The Cu-IR783 sonosensitizer prepared by the present invention is placed in different pH buffer solutions of pH 6.0, pH 6.5 and pH 7.4 for dialyzation for 7 days; TMB is used as a probe to detect the ability of the Cu-IR783 sonosensitizer to generate hydroxyl radicals in a buffer solution containing H2O2.
[0053] The TMB probe is 3, 3', 5, 5'-tetramethylbenzidine probe, and the hydroxyl radical king is •OH.
[0054] Depend on Figure 6 It can be seen that in an acidic environment, the sonodynamic activity of the Cu-IR783 smart sonosensitizer is reactivated and can generate 1 O2 and consumes GSH.
[0055] Based on the above-mentioned Cu-IR783 sonosensitizer, a use of a tumor microenvironment activating sonosensitizer is provided. The Cu-IR783 sonosensitizer prepared above is taken to test the in vitro anti-tumor performance of the Cu-IR783 sonosensitizer, and the survival rate of brain glioma cells treated with the Cu-IR783 sonosensitizer and ultrasonic irradiation is detected by the MTT method.
[0056] The brain glioma cells were seeded in a 96-well plate at 5000 cells / well. After incubation overnight, the old culture medium was replaced with fresh culture medium containing different concentrations of the Cu-IR783 sonosensitizer, and the culture was continued for 24 or 48 hours. At the 4th hour, the cells were ultrasonically treated. Then, the in vitro anti-tumor efficacy of the Cu-IR783 sonosensitizer was detected according to the standard MTT protocol.
[0057] The ultrasonic treatment conditions were 50 kHz, 1.0 W / cm 2 , 5 min.
[0058] Depend on Figure 7 As shown in a, the Cu-IR783 intelligent sonosensitizer cannot release IR783 in LO2 normal cells, but releases a large amount of IR783 in U87-MG cells. Figure 7 As shown in Figure b, at a concentration of 200 μg / mL, no cytotoxicity of Cu-IR783 intelligent sonosensitizer on LO2 cells was detected, indicating that Cu-IR783 intelligent sonosensitizer has good biocompatibility. In contrast, the cell viability of U87-MG cells treated with the same concentration of Cu-IR783 intelligent sonosensitizer decreased to about 45%, indicating that Cu-IR783 intelligent sonosensitizer has tumor cell-specific cytotoxicity. Under US irradiation, the cytotoxic effect of Cu-IR783 intelligent sonosensitizer on cancer cells was further enhanced, indicating that IR783 can be released from Cu-IR783 intelligent sonosensitizer to exert sonodynamic activity.
[0059] Based on the above-mentioned Cu-IR783 sonosensitizer, a use of a tumor microenvironment-activated sonosensitizer is provided, and the above-prepared Cu-IR783 sonosensitizer is used to perform an in vivo anti-tumor performance test.
[0060] a) 2×10 5 U87-MG cells were implanted into the striatum of 4-6 week-old Balb / c nude mice to construct an orthotopic implantable glioma model. Five days after implantation, Cu-IR783 intelligent sonosensitizer was injected through the tail vein, and the mouse brain was ultrasonically treated for 5 minutes. During this period, luciferase was injected intraperitoneally and the growth of tumor cells in the mouse brain was detected using an in vivo bioluminescence / fluorescence imaging system to evaluate the in vivo anti-tumor therapeutic effect of Cu-IR783 intelligent sonosensitizer.
[0061] Depend on Figure 8It can be seen that during the treatment period, the bioluminescent signal of the mouse brain in the control group increased over time, indicating that tumor growth was not restricted. In contrast, the bioluminescent signal of the mouse brain treated with Cu-IR783 intelligent sonosensitizer injection and ultrasound irradiation was much smaller than that of the other groups, indicating that Cu-IR783 intelligent sonosensitizer-mediated SDT, CDT and copper death can effectively treat brain glioma.
[0062] b) To construct a bilateral tumor model, 4-6 week-old Balb / c nude mice were injected with 10 6 4T1 mouse breast cancer cells were used to construct primary tumors. Seven days later, 10 5 4T1 cells were used to establish distal tumors. When the primary tumor volume reached 100 mm 2 After that, the tumor-bearing mice were injected with Cu-IR783 intelligent sonosensitizer and irradiated with ultrasound (50 kHz, 1.0 W / cm 2 , 5 min) treatment, during which the changes in tumor volume of mice were regularly monitored to evaluate the in vivo anti-tumor therapeutic effect of Cu-IR783 intelligent sonosensitizer.
[0063] Depend on Fig. 9 It can be seen that under ultrasound irradiation, the Cu-IR783 intelligent sonosensitizer can completely eradicate primary tumors and effectively induce ex vivo effects to inhibit the growth of distal tumors.
[0064] The Cu-IR783 sonosensitizer prepared in this example is a specially designed and synthesized substance with potential application value. From the perspective of components: IR783 is a near-infrared fluorescent dye with unique optical properties and strong absorption and emission capabilities in the near-infrared region. Near-infrared light has good penetration into biological tissues, which can make IR783-based materials have advantages in fields such as biological imaging. Copper ions can interact with IR783 dyes, and this interaction may change the optical properties, stability or other physical and chemical properties of the dye, thereby giving the material special functions and properties.
[0065] Cu-IR783 has potential application value as a sonosensitizer in cancer treatment. Due to its near-infrared characteristics and responsiveness to ultrasound, the Cu-IR783 intelligent sonosensitizer can accumulate at the tumor site and then be activated by ultrasound irradiation. The reactive oxygen species and other substances produced can destroy the structure and function of tumor cells and achieve the purpose of treating tumors. In addition, the good tissue penetration of near-infrared light enables the sonosensitizer to play a role in tumor treatment in deep tissues.
[0066] In tumor treatment, Cu-IR783 intelligent sonosensitizer has the following advantages: 1. Good tissue penetration: IR783 in the Cu-IR783 intelligent sonosensitizer is a near-infrared fluorescent dye, and near-infrared light has good tissue penetration. This allows the sonosensitizer to penetrate deep into the tumor site in the deep tissue and effectively treat tumors that are difficult to reach with traditional treatments. Compared with other treatments with shorter wavelengths of light excitation, the Cu-IR783 intelligent sonosensitizer can act on deeper tumor tissues, expanding the scope of treatment.
[0067] 2. Accurately locate tumors: 1. Near-infrared fluorescence properties: The near-infrared fluorescence properties of the Cu-IR783 intelligent sonosensitizer can generate strong fluorescence signals in vivo, and through specific imaging equipment, can clearly show the location and size of the tumor. This helps doctors accurately determine the scope of the tumor and provides an important basis for formulating personalized treatment plans.
[0068] 2. Aggregation: Cu-IR783 intelligent sonosensitizer can selectively aggregate at the tumor site. This is because the vascular structure and metabolic characteristics of tumor tissue are different from those of normal tissue, making it easier for Cu-IR783 to accumulate at the tumor site. This selective aggregation improves the targetedness of treatment and reduces damage to normal tissue.
[0069] 3. Low side effects: 1. Small impact on normal tissues: Since the Cu-IR783 intelligent sonosensitizer mainly accumulates at the tumor site and exerts a therapeutic effect under the action of ultrasound, the impact on the surrounding normal tissues is relatively small. Compared with traditional chemotherapy, radiotherapy and other treatment methods, it reduces systemic side effects caused by treatment, such as hair loss, nausea, vomiting, and decreased immunity.
[0070] 2. Good biocompatibility: The ingredients in the Cu-IR783 intelligent sonosensitizer are rationally designed and have good biocompatibility. It will not cause strong immune responses or other adverse biological reactions in the body, reducing the risks during treatment.
[0071] 4. Efficient tumor killing ability 1. Reactive oxygen production: Under the action of ultrasound, Cu-IR783 intelligent sonosensitizer can produce reactive oxygen species (ROS) and other active substances. These active substances have high reactivity and can destroy the structure and function of tumor cells, such as damaging cell membranes, destroying organelles, inducing cell apoptosis, etc., thereby achieving efficient killing of tumor cells.
[0072] 2. Synergistic effect: The synergistic effect of Cu-IR783 and ultrasound can enhance the therapeutic effect. Ultrasound can not only activate sonosensitizers to produce reactive oxygen species, but also directly or indirectly damage tumor cells through mechanical effects, thermal effects and other methods, complementing the effects of sonosensitizers and improving the overall effect of tumor treatment.
[0073] 5. Repeatability and controllability: 1. Repeatable treatment: Cu-IR783 intelligent sonosensitizer can be used multiple times during the treatment process according to the condition of the tumor. Due to its small side effects, patients can withstand multiple treatments, which improves the success rate of tumor treatment and the survival rate of patients.
[0074] 2. Controllable treatment parameters: By adjusting the parameters of ultrasound, such as frequency, intensity, irradiation time, etc., the amount of active oxygen produced by Cu-IR783 and the degree of treatment can be precisely controlled. This allows doctors to make personalized treatment adjustments based on the patient's specific situation and the tumor's response, improving the safety and effectiveness of treatment.
[0075] The above are only specific embodiments of the present invention, but the technical features of the present invention are not limited thereto. Any simple changes, equivalent replacements or modifications made based on the present invention to solve basically the same technical problems and achieve basically the same technical effects are all included in the protection scope of the present invention.
Claims
1. A tumor microenvironment activation sonosensitizer, characterized in that: The invention comprises copper ions and IR783 near-infrared dye, and the copper ions and IR783 near-infrared dye are coordinated and assembled to obtain a Cu-IR783 sonosensitizer of nanoparticles.
2. A method for preparing a tumor microenvironment activated sonosensitizer as claimed in claim 1, characterized in that The following steps are involved: S1. adding the copper ions and IR783 near-infrared dye to a phosphate buffer solution, stirring at room temperature for 20-30 h to obtain a mixed solution, wherein the mass ratio of the copper ions, IR783 near-infrared dye and phosphate buffer solution is 1: (3.5-4.5): (95-105); S2. The mixed solution in step S1 is centrifuged and washed with deionized water for 2-5 times to obtain pure nanoparticle Cu-IR783 sonosensitizer.
3. A use of a sonosensitizer for activating a tumor microenvironment, characterized in that: Take the Cu-IR783 sonosensitizer prepared in claim 2, test the sonodynamic activity of the Cu-IR783 sonosensitizer, and use DPBF as 1 O2 probe, detects the Cu-IR783 sonosensitizer produced under ultrasonic irradiation 1 The levels of O2 were compared with the same concentration of IR783 near-infrared dye to verify the passivation of its sonodynamic activity.
4. A use of a sonosensitizer for activating a tumor microenvironment, characterized in that: The Cu-IR783 sonosensitizer prepared according to claim 2 is taken to test the glutathione consumption of the Cu-IR783 sonosensitizer, and the ability of the IR783 near-infrared dye and the Cu-IR783 sonosensitizer to consume GSH under ultrasonic irradiation is evaluated by using DTNB as a GSH probe.
5. A use of a sonosensitizer for activating a tumor microenvironment, characterized in that: Take the Cu-IR783 sonosensitizer prepared in claim 2, and test the near-infrared imaging capability of the Cu-IR783 sonosensitizer. The fluorescence intensity of the IR783 near-infrared dye and the Cu-IR783 sonosensitizer at the optimal emission peak was detected using a fluorescence spectrometer to evaluate their near-infrared fluorescence quenching.
6. A use of a sonosensitizer for activating a tumor microenvironment, characterized in that: Take the Cu-IR783 sonosensitizer prepared in claim 2, and test the near-infrared imaging capability of the Cu-IR783 sonosensitizer. The near-infrared fluorescence changes of the IR783 near-infrared dye and the Cu-IR783 sonosensitizer are detected by an in vivo imaging monitoring system to evaluate their fluorescence imaging capabilities.
7. A use of a sonosensitizer for activating a tumor microenvironment, characterized in that: The Cu-IR783 sonosensitizer prepared in claim 2 is taken, and the acidic TME response decomposition behavior of the Cu-IR783 sonosensitizer is tested. The Cu-IR783 sonosensitizer prepared by the present invention is placed in different pH buffer solutions of pH 6.0, pH 6.5 and pH 7.4 for dialysis for 7 days; the Cu-IR783 sonosensitizer after dissociation every day is measured, and the absorption spectrum and fluorescence spectrum of the Cu-IR783 sonosensitizer are measured to evaluate the dissociation behavior of the Cu-IR783 sonosensitizer.
8. A use of a sonosensitizer for activating a tumor microenvironment, characterized in that: The Cu-IR783 sonosensitizer prepared in claim 2 is taken, and the acidic TME response decomposition behavior of the Cu-IR783 sonosensitizer is tested. The Cu-IR783 sonosensitizer prepared by the present invention was placed in different pH buffer solutions of pH 6.0, pH 6.5 and pH 7.4 for dialysis for 7 days; 1 The O2 probe and the GSH probe are used to detect the sonodynamic activity and GSH consumption ability of the dissociated Cu-IR783 sonosensitizer.
9. A use of a sonosensitizer for activating a tumor microenvironment, characterized in that: The Cu-IR783 sonosensitizer prepared in claim 2 is taken, and the acidic TME response decomposition behavior of the Cu-IR783 sonosensitizer is tested. The Cu-IR783 sonosensitizer prepared by the present invention was placed in different pH buffer solutions of pH 6.0, pH 6.5 and pH 7.4 for dialysis for 7 days; TMB was used as a probe to detect the ability of the Cu-IR783 sonosensitizer to generate hydroxyl radicals in a buffer solution containing H2O2.
10. A use of a sonosensitizer for activating a tumor microenvironment, characterized in that: Taking the Cu-IR783 sonosensitizer prepared in claim 2, testing the in vitro anti-tumor performance of the Cu-IR783 sonosensitizer, and detecting the survival rate of brain glioma cells treated with the Cu-IR783 sonosensitizer and ultrasonic irradiation by MTT method; The brain glioma cells were seeded in a 96-well plate at 5000 cells / well. After incubation overnight, the old culture medium was replaced with fresh culture medium containing different concentrations of the Cu-IR783 sonosensitizer, and the culture was continued for 24 or 48 hours. At the 4th hour, the cells were ultrasonically treated. Then, the in vitro anti-tumor efficacy of the Cu-IR783 sonosensitizer was detected according to the standard MTT protocol.