Active oxygen response type tumor diagnosis and treatment integrated microneedle patch as well as preparation method and application thereof

Abstract

The invention discloses an active oxygen response type tumor diagnosis and treatment integrated microneedle patch and a preparation method and application thereof. The microneedle patch comprises a microneedle; the active oxygen response type prodrug FDOCl-19 comprises a phenothiazine structure, and the active oxygen response type prodrug FDOCl-19 is loaded on the microneedle. In a microenvironment with high ROS expression in a tumor area, FDOCl-19 is activated to release a large amount of anti-cancer drug melphalan and fluorophore methylene blue in a short time, melphalan effectively prevents growth of tumors, methylene blue can realize in-situ fluorescence imaging of the tumors, and the microneedle patch has the characteristics of on-demand drug delivery and high safety, and can be applied to the field of tumor treatment. And painless, noninvasive and precise treatment of superficial tumors is realized.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine, in particular to a reactive oxygen species (ROS)-responsive tumor diagnosis and treatment integrated microneedle patch, a preparation method and application thereof. BACKGROUND

[0002] Tumor is a malignant disease that seriously threatens human health, and its pathogenesis is complex and has high heterogeneity and invasiveness. As one of the important means of current tumor treatment, chemotherapy mainly relies on the selective killing of cytotoxic drugs on rapidly proliferating tumor cells. However, there are several key problems in the clinical application of traditional chemotherapy, especially in the treatment of superficial tumors (such as skin melanoma), and the efficacy and safety still face great challenges.

[0003] On the one hand, traditional chemotherapy is mostly administered systemically (such as intravenous injection), and the drug is distributed non-specifically in the body, resulting in a large amount of drug accumulation in non-target tissues, inducing serious systemic toxic side effects such as immunosuppression, liver and kidney damage, etc. In addition, due to the particularity of the tumor microenvironment (such as hypoxia, high ROS, etc.), the efficacy of conventional chemotherapy drugs in the lesion area is limited, the treatment response is greatly different, and drug resistance is easily produced. On the other hand, superficial tumors are often supplemented with surgical resection and local injection for local treatment, but surgery has the problems of large trauma, slow postoperative recovery and high tumor recurrence rate; and local injection has the risk of drug diffusion to the surrounding normal tissues, causing non-target toxicity, which limits its clinical promotion. SUMMARY

[0004] The purpose of the present application is to overcome the defects of the existing drugs and treatment methods for poor treatment of superficial tumors.

[0005] In order to achieve the above purpose, the present application provides a ROS-responsive tumor diagnosis and treatment integrated microneedle patch, which comprises: microneedles; and a ROS-responsive prodrug FDOCl-19 containing a phenothiazine structure, which is loaded on the microneedles.

[0006] Optionally, under the action of ROS, the ROS-responsive prodrug FDOCl-19 releases anticancer drug melphalan and fluorescent group methylene blue.

[0007] The present application also provides a preparation method of a ROS-responsive tumor diagnosis and treatment integrated microneedle patch, which comprises at least the following steps:

[0008] Step one, dissolving FDOCl-19 in dimethyl sulfoxide, and ultrasonic dissolving to obtain FDOCl-19 solution;

[0009] Step two, dissolving polylactic acid-glycolic acid copolymer in 1,4-dioxane to obtain a PLGA polymer solution, dissolving polyvinylpyrrolidone in 1,4-dioxane to obtain a PVP polymer solution, mixing the PLGA polymer solution and the PVP polymer solution uniformly to obtain a PLGA / PVP polymer solution;

[0010] Step three, mixing the PLGA / PVP polymer solution and the FDOCl-19 solution uniformly to obtain a microneedle tip solution, adding the microneedle tip solution into a microneedle mold and vacuum drying to make the microneedle tip solution fill the cavities of the microneedle mold and make the solvent evaporate;

[0011] Step four, demolding the FDOCl-19 loaded microneedle patch to obtain an FDOCl-19@MN microneedle patch.

[0012] Optionally, in step two, the monomer polylactic acid: glycolic acid in the polylactic acid-glycolic acid copolymer is (1:1) to (3:1) in molar ratio.

[0013] Optionally, in step two, the PLGA polymer solution:PVP polymer solution is 100:(0-10) in volume ratio.

[0014] Optionally, in step three, the amount of the FDOCl-19 solution is 100-500 μL.

[0015] Optionally, in step three, the vacuum drying time is 10-30 min.

[0016] The application further provides a use of the active oxygen responsive tumor diagnosis and treatment integrated microneedle patch in preparation of a drug for treating superficial tumors.

[0017] Optionally, the superficial tumors at least include any one of breast cancer, skin cancer and melanoma.

[0018] The application further provides a use of the active oxygen responsive tumor diagnosis and treatment integrated microneedle patch in tumor fluorescence imaging.

[0019] Compared with the prior art, the application has at least the following beneficial effects:

[0020] (1) The microneedle patch of the application contains microneedles and active oxygen response type prodrug FDOCl-19 containing phenothiazine structure loaded on the microneedles, when the microneedle patch is used for treating superficial tumors, in the initial administration stage, after the FDOCl-19@MN microneedle patch delivers FDOCl-19 to the tumor area, FDOCl-19 is rapidly activated by ROS in the tumor microenvironment, and a large amount of anticancer drug melphalan and fluorescent group methylene blue is released in a short time, and the growth of the tumor is effectively prevented, and the tumor can be imaged in situ by fluorescence; with the reduction of the tumor, the ROS level in the local area of the tumor decreases, and the release rate of FDOCl-19 decreases, thereby realizing self-adaptive regulation of drug release, and the "on-demand release" mechanism not only helps to maintain the treatment effect, but also effectively avoids the excessive release of drugs in the tumor remission period, and maximally reduces the toxic side effects on normal tissues, which belongs to a dynamic response type intelligent drug delivery mode based on the change of tumor microenvironment.

[0021] (2) The transdermal administration mediated by microneedles can greatly increase the accumulation of drugs in superficial tumors, thereby reducing the drug dosage and maximally reducing the systemic adverse reactions.

[0022] (3) The active oxygen response type prodrug FDOCl-19 shows good stability in the microneedle matrix and can be continuously released for more than 48 days, so that it can realize gradual activation in the tumor microenvironment and realize self-cascading control of intelligent drug release. BRIEF DESCRIPTION OF DRAWINGS

[0023] Figure 1 It is the nuclear magnetic resonance hydrogen spectrum of FDOCl-19 of the application.

[0024] Figure 2 It is the high resolution mass spectrum of FDOCl-19 of the application.

[0025] Figure 3 It is the SEM graph of the FDOCl-19 loaded microneedle patch of the application.

[0026] Figure 4 It is the compression experiment test schematic diagram and stress-displacement curve graph of the FDOCl-19 loaded microneedle patch of the application.

[0027] Figure 5 It is the drug cumulative release rate of the FDOCl-19 loaded microneedle patch (the ratio of PLGA to PVP is 100:0, 100:1 and 100:2) of the application.

[0028] Figure 6 It is the response behavior test schematic diagram, ultraviolet absorption and fluorescence emission spectrum test result graph of the FDOCl-19 loaded microneedle patch of the application.

[0029] Figure 7 Figure 4 is a live fluorescence imaging map of the tumor-bearing mice treated with the FDOCl-19-loaded microneedle patch of the present application.

[0030] Figure 8 Figure 5 is an in vivo anti-tumor treatment effect map of the FDOCl-19-loaded microneedle patch of the present application in a mouse B16 subcutaneous tumor model. DETAILED DESCRIPTION

[0031] The technical solutions of the present application are further described below in combination with the accompanying drawings and examples.

[0032] The experimental materials not mentioned in the present application are commercially available, and the experimental methods not mentioned in the present application are common experimental methods in the art.

[0033] DEFINITIONS

[0034] Chemotherapy

[0035] Chemotherapy is a systemic treatment method that uses chemically synthesized drugs to destroy the growth, proliferation or survival ability of cancer cells. Its core lies in indiscriminately attacking rapidly dividing cells, and is one of the three traditional means of cancer treatment (along with surgery and radiotherapy). The core principle of chemotherapy drugs is to kill rapidly proliferating cancer cells by interfering with the key processes of cell division.

[0036] Chemotherapy Side Effects

[0037] Chemotherapy side effects refer to the unintended damage to normal tissues caused by chemotherapy drugs while killing cancer cells, and the core reason lies in the lack of tumor selectivity of chemotherapy drugs, which indiscriminately attacks normal dividing cells while killing rapidly proliferating cancer cells. The manifestations of chemotherapy side effects vary depending on the type of drug, dose and patient differences, and the high-frequency symptoms mainly include hematological toxicity (incidence > 85%, specific manifestations include neutropenia, thrombocytopenia and anemia, etc.), gastrointestinal toxicity (incidence 70-90%, specific manifestations include nausea and vomiting, mucosal damage and taste abnormalities, etc.), skin and accessory organ damage (alopecia, bullae and nail separation, etc.), neurotoxicity (dose cumulative, specific manifestations include peripheral, autonomic neuropathy and central neurotoxicity, etc.), organ-specific toxicity (heart, lung, kidney and bladder toxicity, etc.), other special toxicity (allergic reactions, secondary tumors and fatigue, etc.).

[0038] Prodrug

[0039] Prodrugs are a class of drug molecules that have been chemically modified to lose their biological activity, and require specific biological transformations (such as enzymatic, metabolic, hydrolytic, etc.) in the body to release the active ingredient to exert therapeutic effects. The core principle of prodrugs is to convert active drugs into inert forms through chemical modification, which can be released under specific conditions in the body to overcome the pharmacokinetic defects, toxicity or delivery barriers of drugs.

[0040] Microneedles (MNs)

[0041] Microneedles are an array of minimally invasive transdermal drug delivery devices with a length of 50-1500 μm and a needle tip diameter of <50 μm, usually made of biocompatible materials (polymers, metals, silicon, etc.), which can penetrate the stratum corneum through physical puncture and form microchannels at the epidermal-dermal junction. The core advantage is to overcome the limitations of traditional drug delivery routes, achieving efficient, precise and painless drug delivery and sustained and precise delivery of drugs / cells.

[0042] In recent years, the prodrug strategy has shown significant advantages in improving the targeting and safety of chemotherapy drugs. Prodrugs are chemical derivatives that can be activated to release the original active ingredient in specific pathological environments in the body, such as enzymatic reactions, pH changes or active oxygen (ROS) enrichment. This strategy not only improves the stability and selective release of drugs, but also effectively reduces the toxic side effects on normal tissues. For example, FDOCl-19 (Release of Amino-orCarboxy-Containing Compounds Triggered by HOCl: Application for Imaging and Drug Design. Angewandte Chemie International Edition 2019, 58(14), 4547-4551) is a ROS-responsive prodrug based on a phenothiazine skeleton. In the tumor microenvironment with high active oxygen levels, FDOCl-19 can be selectively activated to release active drugs with cytotoxicity, accompanied by an increase in fluorescence signal, thereby achieving real-time visualization of the drug release process. This feature provides a theoretical basis and imaging support for precise treatment and dynamic monitoring. However, relying solely on the prodrug strategy still has certain limitations. For example, the enrichment efficiency of drugs at the target site is low, the activation and release process may be limited, the drug efficacy maintenance time is short, and repeated drug administration is required, which is particularly prominent in the treatment of superficial tumors such as melanoma.

[0043] Therefore, developing an efficient, precise and visual local drug delivery system to assist the targeted delivery and responsive release of prodrugs has become an important direction of current research.

[0044] Microneedle patch technology is a new transdermal drug delivery platform, which has the advantages of non-invasive or minimally invasive, easy operation, high compliance, etc. The microneedle structure can effectively penetrate the stratum corneum of the skin and deliver drugs to the epidermis or dermis, thereby bypassing the gastrointestinal metabolism and liver first-pass effect, and significantly improving the bioavailability of drugs. Loading ROS-responsive prodrug FDOCl-19 in the microneedle patch can achieve specific response to local ROS levels in tumors, thereby precisely releasing drugs in the lesion area, and at the same time using its self-contained fluorescent signal for real-time imaging monitoring, constructing a "diagnosis and treatment integrated" treatment strategy. Further, through reasonable design of microneedle morphology, material composition and drug loading method, the accumulation efficiency of drugs in the target tissue can be improved, the therapeutic effect can be enhanced and the impact on healthy tissues can be reduced.

[0045] Therefore, the present application provides a microneedle patch with FDOCl-19 as the core active ingredient, which has ROS responsiveness and fluorescence imaging function. Under the activation of ROS, the anti-cancer drug Melphalan and the fluorescent group Methylene blue (MB) are released, which is used for local precise treatment of superficial tumors.

[0046] The active oxygen-responsive prodrug FDOCl-19 of the present application is synthesized by the inventors' team in the early stage of research, and its synthetic route is shown in formula I, and the specific synthesis steps are described in the literature Chem.Sci., 2018, 9, 495; J. Mater. Chem. B, 2022, 10, 5211-5217. The preparation method is as follows:

[0047] The intermediate acyl chloride FDOCl-2 dissolved in aprotic solvent is added dropwise under ice water bath, and the reaction is carried out under nitrogen after the dropwise addition is completed. The reaction progress is monitored by thin layer chromatography (TLC), and the reaction is continuously stirred until it is completed. After the reaction is completed, the reaction product is extracted with an organic solvent, washed with water, dried with anhydrous sodium sulfate, and evaporated to dryness. The solvent is evaporated to dryness, and the target product FDOCl-19 is obtained by column chromatography. In the preparation stage of compound FDOCl-19, the solvent can be dichloromethane, tetrahydrofuran, or other aprotic solvents. The base can be a carbonate, bicarbonate inorganic salt, or an organic base such as triethylamine, dimethylaminopyridine (DMAP), and the like. The amount of base is calculated in terms of molar amount, and the molar ratio of the acyl chloride is 1:1-5:1. The reaction time is 1-10 h, and the reaction temperature of the system is 0-50°C. In the preparation stage of compound FDOCl-19, the amount of melphalan is calculated in terms of molar amount, and the molar ratio of the acyl chloride is 1:1-5:1. In the extraction stage after the reaction is completed, the amount of organic solvent is 50-200 mL per 1 g of the amount of the material, and FDOCl-19 is finally obtained.

[0048]

[0049] 1. Structure characterization of FDOCl-19

[0050] The compound FDOCl-19 is verified by nuclear magnetic resonance hydrogen spectrum ( Figure 1 ) and high resolution mass spectrum ( Figure 2 ), which is consistent with the literature report, and the results show that the structure of the compound is correct.

[0051] 2. Preparation of microneedle patch

[0052] Step one, dissolve FDOCl-19 in dimethyl sulfoxide (DMSO) and ultrasonicate to dissolve it to obtain FDOCl-19 solution. The amount of dimethyl sulfoxide is 100-500 μL.

[0053] Step two, dissolve polylactic acid-glycolic acid copolymer (PLGA) in 1,4-dioxane to obtain PLGA polymer solution, dissolve polyvinylpyrrolidone in a solvent to obtain PVP polymer solution, and mix the PLGA polymer solution and the PVP polymer solution uniformly to obtain PLGA / PVP polymer solution.

[0054] In some embodiments, the amount of 1,4-dioxane is 0.5-5 mL, and the amount of PLGA is 300-3000 mg. In terms of molar ratio, the monomer polylactic acid: glycolic acid in the polylactic acid-glycolic acid copolymer is (1:1)-(3:1). The viscosity of the polylactic acid-glycolic acid copolymer is 0.1 dl·g-1 ~1.0 dl·g -1 .

[0055] In some embodiments, the PVP has a molecular weight of 4000-7000, the PVP is used in an amount of 300 mg-3000 mg, and the solvent is 1,4-dioxane, which is used in an amount of 0.5 mL-5 mL.

[0056] In some embodiments, the PLGA polymer solution:PVP polymer solution = 100:(1-10) by volume ratio to obtain different ratios of PLGA / PVP polymer solution; the PLGA / PVP polymer solution is used in an amount of 0.5 mL-5 mL.

[0057] Step three, the PLGA / PVP polymer solution is mixed with the FDOCl-19 solution to obtain a microneedle tip solution, the microneedle tip solution is added to the polydimethylsiloxane microneedle mold using a syringe, and vacuum drying is performed to fill the microneedle tip solution into the cavities of the microneedle mold and evaporate the solvent.

[0058] In some embodiments, the FDOCl-19 solution is used in an amount of 100 μL-500 μL; the microneedle tip solution is used in an amount of 100 μL-500 μL.

[0059] Also included is: the PLGA solution is added to the mold groove using a syringe, and then the filled mold is placed in a vacuum drying oven for drying for 10 min-30 min to evaporate the solvent, and the PLGA solution is used in an amount of 0.2 mL-2 mL.

[0060] Step four, after the solvent is completely evaporated, the FDOCl-19-loaded microneedle patch is demolded to obtain the FDOCl-19@MN microneedle patch, which is stored in a -20°C refrigerator in the dark until use.

[0061] A piece of prepared FDOCl-19@MN microneedle patch is ground into powder, 1 mL-10 mL of ethanol is added, and ultrasonic oscillation is performed to prepare the FDOCl-19 microneedle patch extraction solution for subsequent testing.

[0062] 3. Characterization of microneedle patch morphology

[0063] A digital camera (Canon, Japan) is used to take macroscopic morphological characteristics of the FDOCl-19@MN microneedle patch. The FDOCl-19@MN microneedle patch is adhered to the sample stage using conductive adhesive, and gold is sprayed, and then a field emission scanning electron microscope (FE-SEM, S-4800, Hitachi) is used to observe the microscopic morphological characteristics of the FDOCl-19@MN microneedle patch, and the results are as follows Figure 3As shown, a) is a real picture of the microneedle patch, b) is a scanning electron micrograph of the microneedle patch, and the results show that the volume of different PLGA and PVP does not affect the macroscopic and microscopic morphology of the microneedle patch.

[0064] 4. Mechanical strength test of the microneedle patch

[0065] A universal material testing machine (HY-940FS, Hounsfield, UK) was used to test the mechanical strength of the FDOCl-19@MN microneedle patch. Before testing, a piece of microneedle patch was placed on a water platform with the needle tip pointing upwards, the initial distance between the sensor and the needle tip was 1 cm, and the sensor approached the needle tip at a speed of 0.1 mm / s. When the sensor contacted the needle tip, the compression performance was measured, and when the sensor moved 0.6 mm from the needle tip to the backing in the vertical direction, the compression performance measurement was ended, and the stress-displacement relationship curve was drawn by software, as shown in Figure 4 , wherein a) is a test schematic, and b) is a stress-displacement relationship curve.

[0066] 5. In vitro release behavior of FDOCl-19 loaded in the microneedle patch

[0067] Establishment of MB standard curve (dissolved in 50% 1,4-dioxane PBS): A certain amount of MB was weighed, and MB solutions of different concentrations (0.3125, 0.625, 1.25, 2.5, 5 μM) (dissolved in 50% 1,4-dioxane PBS) were prepared. The absorbance was measured by ultraviolet visible spectrophotometer, and the absorbance value was fitted with the concentration to obtain the MB standard curve. Since the microneedle matrix contains components that are not soluble in the water phase, an organic solvent, 1,4-dioxane, is needed to help dissolve the entire microneedle matrix.

[0068] Establishment of MB standard curve (dissolved in 50% ethanol PBS): A certain amount of MB was weighed, and MB solutions of different concentrations (0.3125, 0.625, 1.25, 2.5, 5, 10 μg / mL) (dissolved in 50% ethanol PBS) were prepared. The absorbance was measured by ultraviolet visible spectrophotometer, and the absorbance value was fitted with the concentration to obtain the MB standard curve. This curve is used to determine the drug release kinetics of the microneedle patch. Since the loaded drug FDOCl-19 has poor solubility, ethanol is added to dissolve the released FDOCl-19 from the microneedle patch, while the microneedle matrix is not dissolved. Therefore, the medium system for sustained release is selected as 50% ethanol PBS.

[0069] Determination of drug loading of FDOCl-19@MN microneedle patches: Different MB@MN patches (PLGA:PVP = 100:0, 100:1, and 100:2, V:V) were prepared to estimate the drug loading of different FDOCl-19@MN patches (PLGA:PVP = 100:0, 100:1, and 100:2, V:V). To determine the drug loading of different types of MB@MN patches (PLGA:PVP = 100:0, 100:1, 100:2, V:V), the absorption spectra of MB@MNs were completely dissolved in a mixed solvent (PBS: 1,4-dioxane = 50:50, V / V). The absorbance values ​​(664nm) of each group were then substituted into the MB standard curve (PBS: 1,4-dioxane = 50:50, V / V) to calculate the amount of MB, thereby estimating the drug loading of each group of FDOCl-19@MN microneedle patches.

[0070] The in vitro transdermal release behavior of FDOCl-19@MN microneedle patches was determined on the dorsal skin of mice using a transdermal diffusion apparatus (TP-6, Tianjin Jingtuo Instrument Technology Co., Ltd.). Before the experiment, mouse skin was pretreated by removing hair and subcutaneous fat, and the skin surface was cleaned with absorbent cotton. One FDOCl-19@MN microneedle patch was inserted into the skin and then mounted onto the Franz diffusion apparatus. Throughout the test, the Franz compartment was filled with 8.5 mL of PBS (containing 50% ethanol) and maintained at 37°C with continuous stirring at 80 rpm. At predetermined time intervals, 2 mL of the supernatant was taken and replaced with an equal volume of fresh PBS (containing 50% ethanol). After adding hypochlorous acid (HOCl, 1 mM, 1 μL each time), the MB content in the PBS (containing 50% ethanol) was measured using a UV-Vis spectrophotometer until the absorbance no longer increased. The absorbance values ​​(664 nm) of each group were substituted into the MB standard curve (PBS: ethanol = 50:50, V / V) to estimate the amount of MB in PBS containing 50% ethanol. Then, the MB amount was substituted into the MB release efficiency in HPLC to calculate the cumulative release rate of the FDOCl-19@MN microneedle patch within the preset time interval. The experimental results are as follows: Figure 5 As shown, the larger the PVP volume, the faster the microneedle patch releases the drug, which is beneficial for rapid drug release in the early stages of tumor growth, thereby inhibiting tumor growth.

[0071] 6. Testing the reactive oxygen species response behavior of microneedle patch extract

[0072] like Figure 6 (a) 100 μL of FDOCl-19 microneedle patch extract was added to 2.9 mL of phosphate-buffered saline (PBS, pH 7.4), and HOCl (10, 20, 30, 50, and 100 μM) was added respectively. The fluorescence or absorption spectrum of the solution was detected using a fluorescence or UV spectrophotometer.Figure 6 As shown, the experimental results matched the literature reports, and with the addition of HOCl, the urea bond of FDOCl-19 was broken, releasing methylene blue and melphalan. As shown in b) of Figure 6 , with the release of methylene blue, there was a significant absorption enhancement in the range of 550-700 nm, as shown in c) of Figure 6 , and a significant fluorescence enhancement in the range of 640-800 nm.

[0073] 7. Fluorescent imaging of tumors in tumor-bearing mice

[0074] When the tumor volume of the mice reached 150 mm 3 , the FDOCl-19-loaded microneedle patch was pressed against the tumor surface and remained for about 30 seconds, and then a commercial transparent dressing was used to fix the FDOCl-19-loaded microneedle on the skin surface. Before the fluorescent imaging of the tumor, the FDOCl-19-loaded microneedle patch was removed from the tumor surface, and as shown in Figure 7 , the fluorescent imaging of the tumor was performed at different time points (10, 20, 30, 40, 50, and 60 minutes), and the excitation wavelength was 660 nm at the time of testing.

[0075] 8. Treatment of tumors in tumor-bearing mice

[0076] When the tumor volume of the mice reached 100 mm 3 , the FDOCl-19-loaded microneedle patch was pressed against the tumor surface and remained for about 30 seconds, and then a commercial transparent dressing was used to fix the FDOCl-19-loaded microneedle on the skin surface, and the tumor volume and body weight information of the mice were recorded every two days. As shown in Figure 8 , after the end of the treatment period, the mice were euthanized, and the main organs and tumor tissues were dissected for photography and pathological section analysis Figure 8 ). Among them, A: PBS (i.v., 200 μL, containing 1% DMSO), B: tail vein injection of melphalan (i.v., 500 μM, 200 μL, containing 1% DMSO), C: tail vein injection of FDOCl-19 (i.v., 500 μM, 200 μL, containing 1% DMSO); D: MN (without drug); E: melphalan@MN; F: FDOCl-19@MN, three parallel experiments in each group. It was found that the treatment of group F had the best effect, and the tumor disappeared or was significantly reduced, indicating that the microneedle patch of the application had good efficacy.

[0077] To sum up, the microneedle patch of the present application contains microneedles and active oxygen response prodrug FDOCl-19 containing phenothiazine structure loaded on the microneedles, when the microneedle patch is used for treating superficial tumors, in the initial administration stage, after FDOCl-19@MN microneedle patch delivers FDOCl-19 to the tumor area, FDOCl-19 is rapidly activated by ROS in the tumor microenvironment, and a large amount of anticancer drug melphalan is released in a short time, effectively preventing the growth of tumors, as the tumor shrinks, the ROS level in the local tumor area decreases, and the release rate of FDOCl-19 decreases, thereby realizing self-adaptive regulation of drug release, this "on-demand release" mechanism not only helps to maintain the therapeutic effect, but also effectively avoids the excessive release of drugs in the tumor remission period, maximally reduces the toxic side effects on normal tissues, and belongs to a dynamic response type intelligent drug delivery mode based on the change of tumor microenvironment. Not only has important technical innovation significance, but also shows good clinical transformation potential and broad application prospect.

[0078] Although the content of the present application has been described in detail through the above preferred embodiments, it should be recognized that the above description should not be considered as limiting the present application. After reading the above content, various modifications and alternatives of the present application will be obvious to those skilled in the art. Therefore, the protection scope of the present application should be defined by the appended claims.

Claims

1. A reactive oxygen species-responsive integrated microneedle patch for tumor diagnosis and treatment, characterized in that, The microneedle patch comprises: microneedles; and a reactive oxygen species responsive prodrug FDOCl-19 containing a phenothiazine structure, wherein the reactive oxygen species responsive prodrug FDOCl-19 is loaded onto the microneedles.

2. The microneedle patch as described in claim 1, characterized in that, Under the action of ROS, the reactive oxygen species-responsive prodrug FDOCl-19 is activated to release the anticancer drug melphalan and the fluorophore methylene blue.

3. A method for preparing a reactive oxygen species-responsive tumor diagnostic and therapeutic integrated microneedle patch as described in claim 1 or 2, characterized in that, Include at least the following steps: Step 1: Dissolve FDOCl-19 in dimethyl sulfoxide and sonicate to dissolve it, thus obtaining an FDOCl-19 solution; Step 2: Dissolve polylactic acid-glycolic acid copolymer in 1,4-dioxane to obtain PLGA polymer solution, dissolve polyvinylpyrrolidone in 1,4-dioxane to obtain PVP polymer solution, and mix the PLGA polymer solution and the PVP polymer solution evenly to obtain PLGA / PVP polymer solution. Step 3: Mix the PLGA / PVP polymer solution with the FDOCl-19 solution to obtain a microneedle tip solution. Add the microneedle tip solution to the microneedle mold and then vacuum dry it so that the microneedle tip solution fills the cavity of the microneedle mold and the solvent evaporates. Step 4: Demold the FDOCl-19 loaded microneedle patch to obtain the FDOCl-19@MN microneedle patch.

4. The preparation method according to claim 3, characterized in that, In step two, the molar ratio of polylactic acid to glycolic acid in the polylactic acid-glycolic acid copolymer is (1:1) to (3:1).

5. The preparation method according to claim 3, characterized in that, In step two, the volume ratio of PLGA polymer solution to PVP polymer solution is 100:(0-10).

6. The preparation method according to claim 3, characterized in that, In step three, the amount of FDOCl-19 solution used is 100 μL to 500 μL.

7. The preparation method according to claim 3, characterized in that, In step three, the vacuum drying time is 10 min to 30 min.

8. The application of the reactive oxygen species-responsive tumor diagnostic and therapeutic microneedle patch as described in claim 1 or 2 in the preparation of drugs for treating superficial tumors.

9. The application as described in claim 8, characterized in that, The superficial tumor includes at least one of breast cancer, skin cancer, and melanoma.

10. The application of a reactive oxygen species-responsive tumor diagnostic and therapeutic integrated microneedle patch as described in claim 1 or 2 in tumor fluorescence imaging.

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