Preparation and application of nanoparticle HVC NPs for diagnosis and treatment of Parkinson's disease

Abstract

The invention discloses a preparation method and application of nanoparticles HVC NPs for diagnosis and treatment of Parkinson's disease. The probe HV in the HVC NPs can respond to H2O2 and viscosity, the near-infrared fluorescence imaging capability is played, real-time monitoring of the Parkinson's disease biomarker is achieved, and people are helped to better explore occurrence and development of Parkinson's disease. HVC NPs effectively permeates the blood brain barrier under the assistance of RVG29, curcumin in HVC NPs can relieve apoptosis, reduce the oxidative stress level, eliminate excessive active oxygen, resist inflammation, reduce alpha-syn aggregation and the like, and the effect of effectively resisting Parkinson's disease is shown at the cell and experimental animal level. The nano material provides a new strategy for integrated research on diagnosis and treatment of Parkinson's disease.

Description

Technical Field

[0001] The present invention relates to a preparation method and application of a nano drug for integrated diagnosis and treatment of Parkinson's disease, and in particular to a nano drug modified with a targeting peptide and encapsulated with curcumin, which is a nano material used for targeted penetration of the blood-brain barrier in a Parkinson's disease model and for achieving early diagnosis and treatment. Background Art

[0002] Parkinson's disease (PD) is a common neurodegenerative disease characterized by the loss of dopaminergic neurons in the substantia nigra and a decrease in dopamine content in the striatum. The main clinical manifestations of the disease include resting tremor, muscle rigidity, bradykinesia, and postural instability. Although current treatments can relieve symptoms to a certain extent, they cannot prevent or reverse neuronal degeneration. In addition, existing therapeutic drugs such as levodopa and monoamine oxidase inhibitors may have serious side effects with long-term use, and these drugs are often difficult to cross the blood-brain barrier. The early diagnosis of PD also has the disadvantages of complex methods and low accuracy. Therefore, it is urgent to develop an integrated diagnosis and treatment method to achieve early prevention and treatment of PD and improve the quality of life of patients.

[0003] At present, nanomedicines have shown great potential in the diagnosis and treatment of PD due to their unique physical and chemical properties. Based on our previously synthesized near-infrared organic small molecule fluorescent probe HV, it can be used for imaging of PD models to achieve early diagnosis. However, early diagnosis of abnormalities requires early prevention and treatment to achieve the goal of integrated diagnosis and treatment. In response to the pathogenic factors of PD, such as mitochondrial dysfunction, α-syn aggregation, and inflammatory response, we selected curcumin, an active ingredient of polyphenol drugs, for symptomatic treatment. Summary of the invention

[0004] The purpose of the present invention is to provide a nano drug for integrated diagnosis and treatment of Parkinson's disease and a preparation method thereof. The nano drug is modified with a targeting peptide and encapsulated with curcumin to achieve targeted penetration of the blood-brain barrier, and to perform early diagnosis and treatment of Parkinson's disease models. The preparation method of the nano drug is simple and easy, and has good repeatability and stability. The nano drug effectively penetrates the blood-brain barrier with the assistance of RVG29, and exhibits excellent brain targeting and fluorescence imaging capabilities in a Parkinson's disease animal model. In addition, the nano drug can significantly improve the behavioral disorders of the PD mouse model, increase the number of TH neurons, reduce the occurrence of inflammation, and effectively achieve the treatment of Parkinson's disease and the recovery of physiological functions.

[0005] In order to solve the technical problem of the present invention, the proposed technical solution is: a method for preparing nanoparticles HVC NPs for integrated PD diagnosis and treatment, comprising the following steps:

[0006] (1) Weigh 2 mg of 2-(2-(6-((4-(4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)oxy)-2,3-dihydro-1H-thioanthracenyl-4-yl)vinyl)-1-(2-carboxyethyl)-3,3-dimethyl-3H-indol-1-ium salt (HV) and RVG. 29 -PEG2k-NH2 10mg;

[0007] (2) Place the above substances in a round-bottom flask containing 10 ml of ultrapure water and stir at room temperature;

[0008] (3) adding condensation agents HATU and DIPEA and carrying out condensation reaction for 3 hours to obtain HV-PEG-RVG29;

[0009] (4) dialyzing HV-RVG29 for 24 hours using a 1000 kD dialysis bag to obtain HV-PEG-RV29;

[0010] (5) Using the microemulsion method, 10 mg of the probe HV containing RVG29 and 2 mg of curcumin (C) were dissolved in 1 mL of acetone, respectively, and the mixture was added to 10 mL of deionized water with constant stirring, and the reaction was stirred for 24 hours to finally obtain HVC nanoparticles (HVC NPs);

[0011] The reaction route diagram is as follows:

[0012]

[0013] Preferably, the following are included:

[0014] (1) Add condensing agents HATU and DIPEA to the solution, stir and carry out condensation reaction at room temperature for 3 hours to form a compound HV-RVG29 connected with RVG29. This step is critical to ensure the subsequent biological activity targeting.

[0015] (2) The reaction product HV-RVG29 was dialyzed for 24 hours using a dialysis bag with a molecular weight cutoff of 1000 kD to remove unreacted small molecules and by-products to obtain a pure HV probe containing RVG29.

[0016] In order to solve the technical problem of the present invention, another technical solution is proposed: a nanoparticle HVC NPs, the structure of the nanoparticle HVC NPs includes a probe HV that responds to H2O2 and viscosity, a modified part connected with RVG29-PEG-NH2, and a loaded drug curcumin; RVG29-PEG-NH2 is composed of a targeting peptide RVG29, polyethylene glycol (PEG) and an amino group (-NH2), and is connected to the HV probe to enhance the stability and targeting of the nanoparticles in vivo; through a self-assembly process, nanoparticles with a core-shell structure are formed, wherein the core mainly contains curcumin, and the shell is composed of the HV probe and RVG29-PEG-NH2; the modification of RVG29 enables the nanoparticles to effectively cross the blood-brain barrier, and PD is treated with curcumin.

[0017] In order to solve the technical problem of the present invention, another technical solution is proposed: the application of the nanoparticles HVC NPs for integrated PD diagnosis and treatment, the nanoparticles HVC NPs can be used to prepare nanomedicines that can perform dual detection of H2O2 and viscosity.

[0018] Preferably, nanoparticles HVC NPs can be used to prepare nanomedicines that can restore damage to Parkinson's disease cell models at the cellular level.

[0019] Preferably, the nanoparticles HVC NPs are used to prepare nanomedicines for treating Parkinson's disease.

[0020] Beneficial effects:

[0021] The present invention provides a nanomedicine for integrated diagnosis and treatment of Parkinson's disease, which is modified with a targeting peptide and encapsulated with curcumin, and can penetrate the blood-brain barrier in a targeted manner and achieve early diagnosis and treatment.

[0022] The present invention designs a "diagnosis and treatment integrated" nanoplatform HVC NPs, which forms the final nanomaterial by connecting the targeting peptide RVG29-PEG2k-NH2 and encapsulating the drug curcumin. After tail vein injection, HVC NPs pass through the blood-brain barrier through the action of the targeting peptide and target the acetylcholine receptors on the surface of dopamine neurons. After entering the cell, HVC NPs release the probe HV, which reacts with the H2O2 and viscosity in the cell to increase fluorescence, thereby achieving diagnostic monitoring of the PD process. At the same time, curcumin removes excess reactive oxygen in the cell, restores mitochondrial function, fights inflammation, and restores the number of TH neurons, thereby achieving the goal of integrated diagnosis and treatment in Parkinson's cell models and animal models.

[0023] The present invention obtains a kind of nanoparticle HVC NPs that integrates diagnosis and treatment. The probe HV in HVC NPs can respond to H2O2 and viscosity, exert the ability of near-infrared fluorescence imaging, realize real-time monitoring of Parkinson's disease biomarkers, and help us better explore the occurrence and development of Parkinson's disease. HVC NPs effectively penetrate the blood-brain barrier with the assistance of RVG29. The curcumin in it can reduce cell apoptosis, reduce oxidative stress levels, remove excessive reactive oxygen, fight inflammation, reduce α-syn aggregation, etc., and it has demonstrated effective effects against Parkinson's disease at the cell and experimental animal levels. This nanomaterial provides a new strategy for the integrated diagnosis and treatment of Parkinson's disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 Ultraviolet absorption graph (a) and TEM electron microscope particle size distribution (b) of the nanomedicine HVC NPs of the present invention.

[0025] Figure 2 The ultraviolet-visible absorption spectrum (a) and fluorescence spectrum (b) of the nanomedicine HVC NPs of the present invention under the action of glycerol and H2O2.

[0026] Figure 3 The nanomedicine HVC NPs of the present invention has the ability to respond to both H2O2 and viscosity.

[0027] Figure 4 The nanomedicine HVC NPs of the present invention improves the cell survival rate of the Parkinson's disease cell model induced by rotenone.

[0028] Figure 5 The nanomedicine HVC NPs of the present invention resists cell apoptosis in a Parkinson's disease cell model induced by rotenone.

[0029] Figure 6 The nanomedicine HVC NPs of the present invention showed good brain targeting and fluorescence imaging capabilities in PD model mice.

[0030] Figure 7 The nanomedicine HVC NPs of the present invention improves behavioral disorders in PD model mice.

[0031] Figure 8 The nanomedicine HVC NPs of the present invention increases the number of TH neurons in PD model mice.

[0032] Fig. 9 The nanomedicine HVC NPs of the present invention improves the inflammatory response of PD model mice.

[0033] Fig.10 The nano drug HVC NPs of the invention has good biocompatibility.

[0034] Fig.11 The nanomedicine HVC NPs of the present invention has good biosafety. DETAILED DESCRIPTION

[0035] Example 1

[0036] Synthesis steps of nanomedicine HVC NPs:

[0037] Weigh 2 mg of HV. HV is a patent previously applied for and published by our team. The patent name is a near-infrared probe with dual response to viscosity and hydrogen peroxide and its preparation and application. The publication number is CN117486913A. 1-(2-carboxyethyl)-2-(2-(6-hydroxy-2,3-dihydro-1H-thioxanthene-4-yl)vinyl)-3,3-dimethyl-3H-indol-1-ium salt (100 mg, 0.2 mmol) and Cs2 CO3 (71 mg, 0.2 mmol) were placed in a round-bottom flask, DMF was added, and the reaction was carried out at room temperature for 10 min. Then 2-(4-bromomethylphenyl)-4,4,5,5-tetramethyl-[1,3,2]dioxaborane (59.4 mg, 0.2 mmol) was added to the above mixture, and the reaction was continued for 3 h. After the reaction was completed, the organic phase was washed with saturated brine, the organic phases were combined, and dried over anhydrous Na2 SO4. The solvent was removed and the crude product was purified by silica gel column chromatography (anhydrous methanol: dichloromethane = 1:5, v / v) to obtain a dark blue solid HV.

[0038] 10 mg of RVG29-PEG2k-NH2 was placed in a round-bottom flask with 10 ml of ultrapure water, stirred at room temperature, and then condensation agents HATU and DIPEA were added for 3 hours to obtain HV-RVG29. The mixture was then dialyzed with a 1000 kD dialysis bag for 24 hours to obtain the HV probe containing RVG29.

[0039] HVC NPs were prepared by microemulsion method. 10 mg of probe HV containing RVG29 and 2 mg of curcumin were dissolved in 1 mL of acetone, respectively, and both were added to 10 ml of deionized water with constant stirring, and stirred for 24 h to obtain HVC NPs.

[0040] Example 2

[0041] Ultraviolet absorption spectrum and transmission electron microscopy test of nanomedicine HVC NPs:

[0042] The present invention uses ultraviolet absorption spectroscopy and transmission electron microscopy TEM technology to characterize HVC NPs in detail. Through ultraviolet absorption spectroscopy analysis, HVC NPs show obvious curcumin characteristic absorption peaks at a wavelength of 420nm, and characteristic absorption peaks exclusive to probe HV at a wavelength of 720nm. Figure 1 (a). This result not only confirms the successful loading of curcumin and probe HV, but also indirectly verifies the synthetic accuracy of HVC NPs and the correct ratio of components. Furthermore, through the application of TEM technology, it was observed that HVC NPs have a uniform spherical appearance and a uniform particle size with an average particle size of about 100 nanometers. Figure 1 (b) Based on the above analysis results, it is proved that the synthesis of HVC NPs is successful.

[0043] Example 3

[0044] UV and fluorescence spectra of nanomedicine HVC NPs under the action of H2O2 and glycerol:

[0045] In order to test the properties of HVC nanoparticles as HV probes, 10 μM HVC NPs were added to 2 mL of phosphate buffer solution (PBS), and 100 μM hydrogen peroxide (H2O2) and 4% glycerol were added respectively. After 30 minutes of reaction, its fluorescence and UV absorption were measured. The conditions of the fluorescence test were set to an excitation wavelength of 725 nm, and the excitation and emission slit widths were set to 5 nm and 10 nm, respectively. The results of UV spectral analysis showed that when hydrogen peroxide or glycerol existed alone, the absorbance changed little; however, when both existed at the same time, the absorbance increased significantly, showing an obvious synergistic effect, as shown in Figure 2. Figure 2 (a). Similarly, the fluorescence intensity analysis results show that hydrogen peroxide or glycerol alone has little effect on the fluorescence intensity, but when the two coexist, the fluorescence intensity is significantly enhanced, also showing a synergistic effect, see Figure 2 (b) Based on the above analysis results, HVC NPs have good dual responses to H2O2 and viscosity.

[0046] Example 4

[0047] Cellular Imaging of H2O2 and Viscosity of Nanomedicine HVC NPs in Parkinson's Cell Model:

[0048] In order to study the ability of HVC NPs to dually respond to H2O2 and viscosity at the cellular level, SHSY-5Y cells were seeded in a small dish for confocal microscopy and cultured for 24 hours. Afterwards, the cells were intervened with different drugs: 100μM H2O2 was used to simulate the effects of exogenous hydrogen peroxide, and 10μM rotenone (ROT) was used to simulate endogenous hydrogen peroxide and viscosity changes. After treatment, the cells were washed with PBS, and then fixed with 1ml of paraformaldehyde for 20 minutes, stained with 1ml DAPI for nuclei, and washed three times with PBS. Finally, confocal microscopy was used for cell imaging. The imaging results showed that there was no fluorescence in the blank control group. After adding H2O2, the fluorescence signal increased to a certain extent compared with the blank control group, proving that HVC NPs (10μM) can respond to exogenous H2O2 on cells. In the rotenone-treated group, the red fluorescence signal was stronger. It is proved that HVCNPs can respond to the increased intracellular H2O2 levels and mitochondrial viscosity caused by rotenone, see Figure 3 .

[0049] Example 5

[0050] Nanodrug HVC NPs improve cell survival in Parkinson's cell model:

[0051] In order to evaluate the protective potential of HVC NPs against rotenone-induced SHSY-5Y cell death in the Parkinson's disease (PD) model, we conducted the following experiment. SHSY-5Y cells were first co-treated with 2μM rotenone, and then different concentrations of HVCNPs were added. In the experiment, the concentration of HVC NPs increased from 0μM to 4μM, aiming to evaluate its effect on cell survival. The cell survival rate was determined using the MTT assay, which indirectly reflects the proliferation ability and survival status of cells by measuring mitochondrial activity. The experimental results showed that with the gradual increase in the concentration of HVC NPs, the survival rate of SHSY-5Y cells was significantly improved. In particular, when the concentration of HVC NPs reached 4μM, its effect on anti-rotenone-induced cell apoptosis was most significant. This result shows that HVCNPs can offset the toxic effects of rotenone on SHSY-5Y cells to a certain extent, see Figure 4 .

[0052] Example 6

[0053] Nanomedicine HVC NPs against apoptosis in Parkinson's cell models:

[0054] SHSY-5Y cells were seeded on six-well plates, and the experiment was divided into 4 groups: blank control group, rotenone-induced PD model group, curcumin alone treatment group and HVC NPs treatment group. After treatment with different drugs, the cells were washed three times with PBS. The Calcein-AM mother solution was diluted with PBS to a concentration of 3μM. Subsequently, 1 ml of 3μM Calcein-AM solution was added to each well of cells and incubated for 30 minutes. The cells were washed again and then photographed under a microscope to explore the anti-apoptotic effect of HVC NPs. Through live / dead cell staining, it can be seen that the cells in the control group had better morphology and a larger number. After rotenone treatment, the cells shrank and rounded, and the cell morphology changed. After treatment with curcumin and HVC NPs, the morphology and number were improved. The anti-apoptotic effect of HVCNPs is significantly better than that of the curcumin alone group, as shown in Figure 2. Figure 5 .

[0055] Example 7

[0056] Nanodrug HVC NPs exhibit brain targeting and fluorescence imaging capabilities in Parkinson's disease animal models:

[0057] 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine hydrochloride (MPTP hydrochloride, MPTP) was used to construct a Parkinson's disease mouse model. All mice were 4-6 weeks old and were raised under constant temperature conditions for one week before modeling. MPTP was injected intraperitoneally for 7 consecutive days at a dose of 25 mg / kg. The PD model mice were partially depilated on the head in advance, and HVCNPs were injected into the tail vein of the PD model mice at a dose of 10 mg / kg / time, for a total of 6 injections (once a day), and the fluorescence imaging ability of the mice was observed at different time periods. In PD mice, a fluorescent signal appeared on the head after 3 hours, the strongest fluorescent signal of the mice was 6-9 hours later, the fluorescent signal began to weaken at 12 hours, and some fluorescent signals still existed at 24 hours, proving that HVC NPs have the ability of fluorescence imaging in PD mice, see Figure 6 .

[0058] Example 8

[0059] Nanomedicine HVC NPs improve behavioral disorders in animal models of Parkinson's disease:

[0060] C57 mice aged 4-6 weeks were used as models (MPTP-constructed PD mouse models), and the mice were randomly divided into four groups: control group, MPTP group, MPTP plus curcumin group, and MPTP plus HVC NPs group, with 10 mice in each group. MPTP was injected intraperitoneally at a dose of 25 mg / kg for 7 consecutive days. During this period, HVC NPs and curcumin (10 mg / kg) were injected into the tail vein three days before MPTP injection, the third day after MPTP injection, the sixth day after MPTP injection, and the second day after MPTP injection. HVCNPs were injected into the tail vein at a dose of 10 mg / kg / time, for a total of 6 injections (once a day). The dose was consistent with the dose of curcumin alone, and the tail vein was injected 6 times in total. The mice were placed in a 40*40*40cm box. The crawling trajectory of the mice was collected with a camera and analyzed with software. The mice in the control group walked around randomly in the open field, without a fixed form, had a greater interest in exploring unknown areas, had a longer movement distance, a faster speed, and stayed in the central area for a longer time. The mice in the MPTP group moved slowly, had no interest in exploring unknown areas, walked along the edges, spent less time in the central area, and had a shorter total distance of movement. In the curcumin treatment group, the slow movement caused by MPTP was improved, the total distance of movement increased, and the movement speed also increased slightly. In the HVC NPs treatment group, the total distance of movement was significantly improved compared to the MPTP group, the movement speed was significantly increased, the time spent in the central area was longer, and the interest in exploring unknown areas was also greatly improved. It can be seen that HVC NPs can significantly improve the behavioral damage caused by MPTP. Figure 7 .

[0061] Example 9

[0062] Nanodrug HVC NPs increase the number of TH neurons in animal models of Parkinson's disease:

[0063] Tyrosine hydroxylase (TH) is an important marker of dopaminergic neurons. Therefore, detecting the number of TH neurons can determine the protective effect of HVC NPs on PD model mice caused by MPTP. The brain tissue was taken for paraffin sectioning, and immunohistochemical staining and immunofluorescence staining of TH neurons were performed. The immunohistochemical results showed that the TH neurons in the control group were intact and normal in number, while the number of TH neurons in the MPTP group was significantly reduced. The number of TH neurons in the curcumin treatment group recovered to a certain extent, and the number of TH neurons in the HVC NPs treatment group (10 mg / kg, injected 6 times a day) recovered significantly, which was significantly different from the MPTP group, and there was no significant difference from the control group, see Figure 8 .

[0064] Example 10

[0065] Nanomedicine HVC NPs fights inflammatory response in animal models of Parkinson's disease:

[0066] In the brain inflammatory response of Parkinson's disease, CD206-labeled M2 macrophages play a key role in regulating and alleviating inflammation. These cells help alleviate pathological conditions by promoting inflammatory resolution and tissue repair. To evaluate the effects of different treatments on these cells, we performed immunofluorescence staining on brain tissue sections. The results showed that in the MPTP-induced Parkinson's disease model, the number of CD206-positive macrophages was reduced compared with the control group, indicating that MPTP may inhibit the activity or survival of M2 macrophages. In contrast, the number of CD206-positive macrophages in the curcumin and HVC NPs treatment groups (10 mg / kg injected once a day for 6 times) increased, indicating that these two treatment strategies may exert anti-inflammatory effects by promoting the recovery or activation of M2 macrophages. In particular, the increase in CD206-positive cells in the HVC NPs treatment group exceeded that in the curcumin treatment group, suggesting that HVC NPs may have a stronger anti-inflammatory mechanism mediated by promoting M2 macrophages, see Fig. 9 .

[0067] Embodiment 11

[0068] Nanomedicine HVC NPs has good biocompatibility:

[0069] To evaluate the blood compatibility of HVC NPs with red blood cells, red blood cells were first separated from whole blood and resuspended in PBS to prepare a red blood cell suspension. Next, HVC nanoparticle solutions of different concentrations (0 to 40 μM) were prepared. In the experiment, pure water was set as the positive control group, and PBS was set as the negative control group. After the red blood cell suspension was mixed with the nanoparticle solutions of various concentrations, they were incubated at 37°C for a certain period of time. After the incubation, the unbroken red blood cells and the supernatant containing the hemolyzed product were separated by centrifugation, and the absorbance of hemoglobin was measured at 540 nm using a spectrophotometer to evaluate the degree of hemolysis. The hemolysis rate was calculated, and the data was analyzed to determine the blood compatibility of HVC NPs. The results showed that no obvious hemolysis was observed in the concentration range of 0-40 μM, indicating that HVC NPs have good blood compatibility, see Fig.10 .

[0070] Example 12

[0071] Nanomedicine HVC NPs has good biosafety:

[0072] In order to evaluate the biosafety of the nanomedicine HVC NPs, serum was collected from four groups of mice, including the control group, MPTP group, MPTP plus curcumin group, and MPTP plus HVC NPs group, for blood biochemical analysis. The measurement results of biochemical indices including aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea nitrogen (BUN), and creatinine (CRE) were all within the normal range, see Fig.11 . Furthermore, there were no significant abnormalities in the levels of AST, ALT, BUN and CRE between the different experimental groups, and these indicators reflect the functional status of the liver and kidneys. Among them, AST and ALT are mainly used to evaluate liver damage, while BUN and CRE are important indicators for evaluating kidney function. The stability of indicators in all groups showed that HVC NPs did not cause obvious damage to the liver and kidneys of experimental mice at our experimental dose (10 mg / kg), thus proving its good biosafety. This result not only shows the protection of HVC nanoparticles on key organ functions, but also provides a solid experimental basis for its safety in clinical applications.

Claims

1. A method for preparing nanoparticles HVC NPs for diagnosis and treatment of Parkinson's disease, characterized in that: The steps include: (1) Weigh 2 mg of 2-(2-(6-((4-(4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)oxy)-2,3-dihydro-1H-thioanthracenyl-4-yl)vinyl)-1-(2-carboxyethyl)-3,3-dimethyl-3H-indol-1-ium salt (HV) and RVG. 29 -PEG2k-NH2 10mg; (2) Place the above substances in a round-bottom flask containing 10 ml of ultrapure water and stir at room temperature; (3) adding condensation agents HATU and DIPEA and carrying out condensation reaction for 3 hours to obtain HV-PEG-RVG29; (4) dialyzing HV-RVG29 for 24 hours using a 1000 kD dialysis bag to obtain HV-PEG-RV29; (5) Using the microemulsion method, 10 mg of RVG29-containing probe HV and 2 mg of curcumin (C) were dissolved in 1 mL of acetone, respectively. The mixture was then added to 10 mL of deionized water with constant stirring and reacted for 24 hours to finally obtain HVC nanoparticles (HVC NPs). The reaction route diagram is as follows:

2. The method for preparing nanoparticles HVC NPs for diagnosis and treatment of Parkinson's disease according to claim 1, characterized in that: These include: (1) adding condensing agents HATU and DIPEA to the solution, stirring and carrying out condensation reaction at room temperature for 3 hours to form a compound HV-RVG29 connected with RVG29. This step is critical to ensure subsequent biological activity targeting; (2) The reaction product HV-RVG29 was dialyzed for 24 hours using a dialysis bag with a molecular weight cutoff of 1000 kD to remove unreacted small molecules and by-products to obtain a pure HV probe containing RVG29.

3. A nanoparticle HVC NPs for diagnosis and treatment of Parkinson's disease, characterized in that: The structure of nanoparticles HVC NPs includes a probe HV that responds to H2O2 and viscosity, a modified part connected with RVG29-PEG-NH2, and loaded drug curcumin; RVG29-PEG-NH2 is composed of targeting peptide RVG29, polyethylene glycol (PEG) and amino group (-NH2), and is connected to the HV probe to enhance the stability and targeting of the nanoparticles in vivo; through the self-assembly process, nanoparticles with a core-shell structure are formed, in which the core mainly contains curcumin and the shell is composed of HV probe and RVG29-PEG-NH2; the modification of RVG29 enables the nanoparticles to effectively cross the blood-brain barrier, and use curcumin to treat Parkinson's disease.

4. The use of nanoparticles HVC NPs as claimed in claim 3, characterized in that: Nanoparticles HVC NPs can be used to prepare nanomedicines with dual detection of H2O2 and viscosity.

5. The use of nanoparticles HVC NPs as claimed in claim 2, characterized in that: Nanoparticles HVC NPs can be used to prepare nanomedicines that can restore damage to Parkinson's disease cell models at the cellular level.

6. The use of nanoparticles HVC NPs as claimed in claim 2, characterized in that: Nanoparticles HVC NPs are used to prepare nanomedicines for the treatment of Parkinson's disease.

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