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

Abstract

This invention discloses a method for preparing HVC NPs for the diagnosis and treatment of Parkinson's disease and their applications. The probe HV in the HVC NPs responds to H2O2 and viscosity, utilizing near-infrared fluorescence imaging capabilities to achieve real-time monitoring of Parkinson's disease biomarkers, helping us better understand the occurrence and development of Parkinson's disease. With the assistance of RVG29, the HVC NPs effectively cross the blood-brain barrier. The curcumin within them can reduce cell apoptosis, lower oxidative stress levels, scavenge excess reactive oxygen species, combat inflammation, and reduce α-synuclein aggregation, demonstrating effective anti-Parkinson's disease effects at the cellular and experimental animal levels. This nanomaterial provides a new strategy for integrated research on the diagnosis and treatment of Parkinson's disease.

Description

Technical Field

[0001] This invention relates to a method for preparing an integrated nanomedicine for the diagnosis and treatment of Parkinson's disease and its application. Specifically, it relates to a nanomedicine modified with a targeted peptide and encapsulated with curcumin, which is used in a Parkinson's disease model to target and penetrate the blood-brain barrier for early diagnosis and treatment. Background Technology

[0002] Parkinson's disease (PD) is a common neurodegenerative disorder characterized by the loss of dopaminergic neurons in the substantia nigra and a decrease in dopamine levels in the striatum. The main clinical manifestations include resting tremor, rigidity, bradykinesia, and postural instability. While current treatments can alleviate symptoms to some extent, they cannot stop or reverse neuronal degeneration. Furthermore, existing medications such as levodopa and monoamine oxidase inhibitors may cause serious side effects with long-term use, and these drugs often have difficulty crossing the blood-brain barrier. Early diagnosis of PD also suffers from complex methods and low accuracy. Therefore, there is an urgent need to develop an integrated diagnostic and therapeutic approach to achieve early prevention and treatment of PD and improve patients' quality of life.

[0003] Currently, nanomedicines have shown great potential in the diagnosis and treatment of Parkinson's disease (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 PD models, thereby enabling early diagnosis. However, early diagnosis of abnormalities necessitates early prevention and treatment to achieve the goal of integrated diagnosis and therapy. Targeting the pathogenic factors of PD, such as mitochondrial dysfunction, α-synuclein aggregation, and inflammatory responses, we selected curcumin, an active ingredient in polyphenolic drugs, for symptomatic treatment. Summary of the Invention

[0004] The purpose of this invention is to provide a nanomedicine for the integrated diagnosis and treatment of Parkinson's disease and its preparation method. This nanomedicine, through targeted peptide modification and curcumin encapsulation, achieves targeted penetration of the blood-brain barrier, enabling early diagnosis and treatment of Parkinson's disease models. The preparation method of this nanomedicine is simple and easy to perform, exhibiting good reproducibility and stability. With the assistance of RVG29, the nanomedicine effectively crosses the blood-brain barrier and demonstrates excellent brain targeting and fluorescence imaging capabilities in animal models of Parkinson's disease. Furthermore, this nanomedicine can significantly improve behavioral disorders in PD mouse models, increase the number of TH neurons, and reduce inflammation, effectively achieving the treatment of Parkinson's disease and the recovery of physiological function.

[0005] To address the technical problem of this invention, the proposed technical solution is as follows: a method for preparing HVC NPs for integrated diagnosis and treatment of PD, comprising the following steps:

[0006] (1) Weigh 2 mg of 2-(2-(6-((4-(4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)benzyl)oxy)-2,3-dihydro-1H-thioanthrayl-4-yl)vinyl)-1-(2-carboxyethyl)-3,3-dimethyl-3H-indole-1-onium inner salt (HV) and RVG 29 -PEG2k-NH2 10mg;

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

[0008] (3) Add condensing agents HATU and DIPEA, and carry out condensation reaction for 3 hours to obtain HV-PEG-RVG29;

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

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

[0011] The reaction route diagram is as follows:

[0012]

[0013] Preferred options include the following:

[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 compound HV-RVG29 linked to RVG29. This step is crucial to ensure subsequent bioactivity targeting.

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

[0016] To address the technical problem of this invention, another technical solution is proposed: a nanoparticle HVC NPs, the structure of which includes a probe HV responsive to H2O2 and viscosity, a modified portion connected to RVG29-PEG-NH2, and loaded with the drug curcumin; RVG29-PEG-NH2 is composed of the targeting peptide RVG29, polyethylene glycol (PEG), and amino group (-NH2), and by connecting with the HV probe, the stability and targeting of the nanoparticles in vivo are enhanced; 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 curcumin is used to treat PD.

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

[0018] Preferably, HVC NP nanoparticles can be used to prepare nanomedicines for detecting the recovery of damage in a Parkinson's disease cell model at the cellular level.

[0019] Preferably, HVC NP nanoparticles are used to prepare nanomedicines for the treatment of Parkinson's disease.

[0020] Beneficial effects:

[0021] This invention provides a nanomedicine for the integrated diagnosis and treatment of Parkinson's disease. By modifying and encapsulating curcumin with targeted peptides, it can penetrate the blood-brain barrier and achieve early diagnosis and treatment.

[0022] This invention designs a "therapeutic" 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 cross the blood-brain barrier through the targeting peptide and target acetylcholine receptors on the surface of dopamine neurons. Upon entering the cell, HVC NPs release the probe HV, which reacts with intracellular H2O2 and viscosity, resulting in increased fluorescence and enabling diagnostic monitoring of Parkinson's disease progression. Simultaneously, curcumin scavenges excess reactive oxygen species within the cell, restores mitochondrial function, combats inflammation, and restores the number of TH neurons, thus achieving the goal of therapeutic integration in Parkinson's cell and animal models.

[0023] This invention yields a therapeutically integrated nanoparticle, HVC NPs. The probe HV within the HVC NPs responds to H2O2 and viscosity, utilizing near-infrared fluorescence imaging capabilities to achieve real-time monitoring of Parkinson's disease biomarkers, helping us better understand the occurrence and development of Parkinson's disease. With the assistance of RVG29, the HVC NPs effectively cross the blood-brain barrier. The curcumin within them can reduce cell apoptosis, lower oxidative stress levels, scavenge excess reactive oxygen species, combat inflammation, and reduce α-synuclein aggregation, demonstrating effective anti-Parkinson's disease effects at the cellular and experimental animal levels. This nanomaterial provides a new strategy for integrated diagnostic and therapeutic research on Parkinson's disease. Attached Figure Description

[0024] Figure 1 The ultraviolet absorption spectrum (a) and TEM electron microscopy particle size distribution (b) of the HVC NPs nanomedicines of this invention.

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

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

[0027] Figure 4 The nanomedicine HVC NPs of this invention improves cell survival rate in a rotenone-induced Parkinson's disease cell model.

[0028] Figure 5 The nanomedicine HVC NPs of this invention inhibits apoptosis in a rotenone-induced Parkinson's disease cell model.

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

[0030] Figure 7 The nanomedicine HVC NPs of this invention improve behavioral disorders in PD model mice.

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

[0032] Figure 9 The nanomedicine HVC NPs of this invention improve the inflammatory response in PD model mice.

[0033] Figure 10 The HVC NPs nanomedicines of this invention exhibit good biocompatibility.

[0034] Figure 11 The HVC NPs nanomedicines of this invention have good biosafety. Detailed Implementation

[0035] Example 1

[0036] Synthesis steps of HVC NPs nanomedicines:

[0037] Weigh 2 mg of HV. HV is from a previously applied and published patent of our team, entitled "A Near-Infrared Probe Responding to Both Viscosity and Hydrogen Peroxide and Its Preparation and Application," publication number CN117486913A. 1-(2-Carboxyethyl)-2-(2-(6-hydroxy-2,3-dihydro-1H-thioxanthracene-4-yl)vinyl)-3,3-dimethyl-3H-indole-1-onium inner salt (100 mg, 0.2 mmol) and Cs₂CO₃ (71 mg, 0.2 mmol) were placed in a round-bottom flask, DMF was added, and the mixture was reacted 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 mixture, and the reaction was continued for 3 h. After the reaction was complete, the organic phase was washed with saturated brine, the organic phases were combined, and dried over anhydrous Na₂SO₄. After removing the solvent, 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 containing 10 ml of ultrapure water and stirred at room temperature. Then, condensation agents HATU and DIPEA were added, and the condensation reaction was carried out for 3 hours to obtain HV-RVG29. The mixture was then dialyzed for 24 hours using a 1000 kDa dialysis bag to obtain an HV probe containing RVG29.

[0039] HVC NPs were prepared using a microemulsion method. 10 mg of probe HV containing RVG29 and 2 mg of curcumin were dissolved separately in 1 mL of acetone. Both were then added to 10 mL of deionized water under constant stirring, and the reaction was carried out for 24 h. HVC NPs were obtained.

[0040] Example 2

[0041] UV absorption spectroscopy and transmission electron microscopy of HVC NPs nanomedicines:

[0042] This invention employs ultraviolet absorption spectroscopy and transmission electron microscopy (TEM) techniques to characterize HVC NPs in detail. Ultraviolet absorption spectroscopy analysis revealed that HVC NPs exhibited a distinct curcumin characteristic absorption peak at 420 nm, while displaying a characteristic absorption peak specific to the probe HV at 720 nm. Figure 1 (a) This result not only confirms the successful loading of curcumin with the probe HV, but also indirectly verifies the accuracy of the synthesis of HVC NPs and the correct ratio of components. Furthermore, through the application of TEM technology, it was observed that the HVC NPs have a uniform spherical appearance and a uniform particle size, with an average particle size of approximately 100 nanometers. Figure 1 (b). Based on the above analysis results, the synthesis of HVC NPs was successfully demonstrated.

[0043] Example 3

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

[0045] To test the properties of HVC nanoparticles as HV probes, 10 μM HVC NPs were added to 2 mL of phosphate-buffered saline (PBS), followed by the addition of 100 μM hydrogen peroxide (H₂O₂) and 4% glycerol. After 30 minutes of reaction, fluorescence and UV absorption were measured. Fluorescence testing was performed with an excitation wavelength of 725 nm and excitation and emission slit widths of 5 nm and 10 nm, respectively. UV spectroscopy analysis showed that absorbance changes were minimal when hydrogen peroxide or glycerol were present alone; however, a significant increase in absorbance was observed when both were present simultaneously, exhibiting a clear synergistic effect (see [reference needed]). Figure 2 (a). Similarly, fluorescence intensity analysis showed that hydrogen peroxide or glycerol alone had little effect on fluorescence intensity, but when both coexisted, the fluorescence intensity was significantly enhanced, also showing a synergistic effect, see [reference needed]. Figure 2 (b). Based on the above analysis results, HVC NPs exhibit a good dual response to H2O2 and viscosity.

[0046] Example 4

[0047] Cellular imaging of H2O2 and viscosity of nanomedicine HVC NPs in a Parkinson's cell model:

[0048] To investigate the dual cellular response of HVC NPs to H2O2 and viscosity, SHSY-5Y cells were seeded in confocal microscope dishes and cultured for 24 hours. Subsequently, the cells were treated with different drugs: 100 μM H2O2 was used to simulate the effect of exogenous hydrogen peroxide, and 10 μM rotenone (ROT) was used to simulate endogenous hydrogen peroxide and viscosity changes. After treatment, the cells were washed with PBS, fixed with 1 ml paraformaldehyde for 20 minutes, stained with 1 ml DAPI, and washed three times with PBS. Finally, cell imaging was performed using confocal microscopy. The imaging results showed that the blank control group had no fluorescence, while the fluorescence signal increased to some extent after the addition of H2O2 compared to the blank control group, demonstrating that HVC NPs (10 μM) can respond to exogenous H2O2 on cells. In the rotenone-treated group, the red fluorescence signal was stronger. This demonstrates that HVC NPs can respond to the increase in intracellular H2O2 levels and mitochondrial viscosity induced by rotenone. Figure 3 .

[0049] Example 5

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

[0051] To evaluate the protective potential of HVC NPs against rotenone-induced apoptosis in a Parkinson's disease (PD) model of SHSY-5Y cells, we conducted the following experiment: SHSY-5Y cells were first co-treated with 2 μM rotenone, followed by the addition of different concentrations of HVC NPs. The concentration of HVC NPs was increased from 0 μM to 4 μM to assess its effect on cell viability. Cell viability was determined using the MTT assay, which indirectly reflects cell proliferation and survival status by measuring mitochondrial activity. The results showed that the viability of SHSY-5Y cells significantly increased with gradually increasing HVC NP concentration. Particularly at a concentration of 4 μM, the anti-rotenone-induced apoptosis effect was most significant. This result indicates that HVC NPs can, to some extent, counteract the toxic effects of rotenone on SHSY-5Y cells. Figure 4 .

[0052] Example 6

[0053] Nanomedicine HVC NPs combat apoptosis in a Parkinson's cell model:

[0054] SHSY-5Y cells were seeded into six-well plates, and the experiment was divided into four groups: a blank control group, a rotenone-induced PD model group, a curcumin-only treatment group, and an HVC NPs treatment group. After treatment with different drugs, the cells were washed three times with PBS. Calcein-AM stock solution was diluted with PBS to a concentration of 3 μM. Subsequently, 1 ml of 3 μM Calcein-AM solution was added to each well of cells, and the cells were incubated for 30 minutes. After washing the cells again, the cells were photographed under a microscope to explore the anti-apoptotic effect of HVC NPs. Live / dead cell staining showed that the control group had better cell morphology and a larger number of cells. After rotenone treatment, the cells shrank and became rounded, and the cell morphology changed. After curcumin treatment and HVC NPs treatment, both morphology and number improved. The anti-apoptotic effect of HVC NPs was significantly better than that of the curcumin-only group. Figure 5 .

[0055] Example 7

[0056] The nanomedicine HVC NPs demonstrated brain targeting and fluorescence imaging capabilities in an animal model of Parkinson's disease:

[0057] A mouse model of Parkinson's disease (PD) was established using 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine hydrochloride (MPTP hydrochloride, MPTP). All mice were 4-6 weeks old and induced into the PD model after one week of homeostasis. The mice were intraperitoneally injected with MPTP at a dose of 25 mg / kg for 7 consecutive days. Hair was removed from the head of the PD model mice beforehand. HVCNPs were injected via the tail vein at a dose of 10 mg / kg per injection, for a total of 6 injections (once a day). The fluorescence imaging ability of the mice was observed at different time points. In PD mice, fluorescence signals appeared on the head after 3 hours, with the strongest fluorescence signals at 6-9 hours. The fluorescence signals began to weaken at 12 hours, and some fluorescence signals were still present at 24 hours, demonstrating that HVC NPs have the ability to produce fluorescence imaging in PD mice. (See below) 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 a PD mouse model (MPTP-based PD mouse model). 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 administered intraperitoneally at a dose of 25 mg / kg for 7 consecutive days. During this period, HVC NPs and curcumin (10 mg / kg) were injected via tail vein on the first three days, the third and sixth days after MPTP injection, and the second day after the end of MPTP injection. HVC NPs were injected via tail vein at a dose of 10 mg / kg per injection, for a total of 6 injections (once a day), the same dose as curcumin alone, for a total of 6 tail vein injections. Mice were placed in a 40*40*40cm box. The mice's crawling trajectories were collected using a camera and analyzed using software. The control group mice wandered randomly in the open field, exhibiting no fixed pattern, showing greater interest in exploring unknown areas, traveling longer distances, moving faster, and spending more time in the central area. Mice in the MPTP group exhibited sluggish movement, lacked interest in exploring unknown territories, walked along the edges, spent less time in the central area, and covered a shorter total distance. In the curcumin treatment group, MPTP-induced sluggishness was improved, total distance covered increased, and movement speed slightly improved. In the HVC NPs treatment group, total distance covered was significantly improved compared to the MPTP group, movement speed was significantly increased, time spent in the central area was longer, and interest in exploring unknown areas was significantly enhanced. Therefore, HVC NPs can significantly improve the behavioral impairments caused by MPTP. Figure 7 .

[0061] Example 9

[0062] Nanomedicine 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 against MPTP-induced PD model mice. Brain tissue was sectioned in paraffin and subjected to immunohistochemical and immunofluorescence staining for TH neurons. Immunohistochemical results showed that the control group had intact and normal TH neurons, while the MPTP group showed a significant reduction in the number of TH neurons. The curcumin treatment group showed some recovery in the number of TH neurons, and the HVC NPs treatment group (10 mg / kg, injected 6 times daily) showed significant recovery in the number of TH neurons, significantly different from the MPTP group but not significantly different from the control group. Figure 8 .

[0064] Example 10

[0065] Nanomedicine HVC NPs against inflammatory response in an animal model of Parkinson's disease:

[0066] In the inflammatory response of the brain in Parkinson's disease, CD206-labeled M2 macrophages play a crucial role in regulating and alleviating inflammation. These cells contribute to mitigating the pathological condition by promoting inflammation resolution and tissue repair. To assess the effects of different treatments on these cells, we performed immunofluorescence staining on brain tissue sections. The results showed that in an MPTP-induced Parkinson's disease model, the number of CD206-positive macrophages was reduced compared to the control group, suggesting that MPTP may inhibit the activity or survival of M2 macrophages. In contrast, the number of CD206-positive macrophages increased in both the curcumin and HVC NPs treatment groups (10 mg / kg, 6 injections once daily), 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 was greater than that in the curcumin treatment group, suggesting that HVC NPs may have a stronger mechanism for promoting M2 macrophage-mediated anti-inflammatory effects. Figure 9 .

[0067] Example 11

[0068] The nanomedicine HVC NPs exhibits good biocompatibility.

[0069] To assess the blood compatibility of HVC NPs with erythrocytes, erythrocytes were first isolated from whole blood and resuspended in PBS to prepare erythrocyte suspensions. Next, HVC nanoparticle solutions of different concentrations (0–40 μM) were prepared. Pure water was used as a positive control, and PBS as a negative control. The erythrocyte suspensions were mixed with the nanoparticle solutions of each concentration and incubated at 37°C for a certain period. After incubation, unruptured erythrocytes were separated from the supernatant containing hemolytic products by centrifugation. The absorbance of hemoglobin was measured at 540 nm using a spectrophotometer to assess the degree of hemolysis. The hemolysis rate was calculated, and the data were analyzed to determine the blood compatibility of HVC NPs. The results showed that no significant hemolysis was observed within the concentration range of 0–40 μM, indicating that HVC NPs have good blood compatibility. Figure 10 .

[0070] Example 12

[0071] The nanomedicine HVC NPs exhibits good biosafety.

[0072] To assess the biosafety of HVC nanoparticles, serum samples were collected from four groups of mice: a control group, an MPTP group, an MPTP plus curcumin group, and an MPTP plus HVC NPs group, for blood biochemical analysis. The biochemical parameters, including aspartate aminotransferase (AST), alanine aminotransferase (ALT), blood urea nitrogen (BUN), and creatinine (CRE), were all within the normal range. Figure 11 Furthermore, no significant abnormalities were observed in the levels of AST, ALT, BUN, and CRE across the different experimental groups. These indicators reflect the functional status of the liver and kidneys. AST and ALT are primarily used to assess liver damage, while BUN and CRE are important indicators for assessing kidney function. The stability of these indicators across all groups indicates that HVC NPs did not cause significant damage to the liver and kidneys of experimental mice at our experimental dose (10 mg / kg), thus demonstrating their good biocompatibility. This result not only demonstrates the protective effect of HVC nanoparticles on the function of critical organs but also provides a solid experimental basis for their safety in clinical applications.

Claims

1. A method for preparing HVC NPs nanoparticles for the diagnosis and treatment of Parkinson's disease, characterized in that, Includes the following steps: (1) Weigh 2 mg of 2-(2-(6-((4-(4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)benzyl)oxy)-2,3-dihydro-1H-thioanthrayl-4-yl)vinyl)-1-(2-carboxyethyl)-3,3-dimethyl-3H-indole-1-onium inner salt (HV) and RVG 29 -PEG2k-NH2 10 mg; (2) Place the above substances into a round-bottom flask containing 10 ml of ultrapure water and stir at room temperature; (3) Add condensing agents HATU and DIPEA, and carry out condensation reaction for 3 hours to obtain HV-PEG-RVG. 29 ; (4) HV-PEG-RVG29 was dialyzed for 24 hours using a 1000 kD dialysis bag to obtain HV-PEG-RVG29. 29 HV probe; (5) Using the microemulsion method, 10 mg of RVG-containing 29 The probe HV and 2 mg of curcumin (C) were dissolved in 1 mL of acetone, and the mixture was then added to 10 mL of deionized water under constant stirring. The mixture was stirred for 24 hours to finally obtain HVC nanoparticles (HVC NPs). The reaction route diagram is as follows: 。 2. The method for preparing HVC NPs for the diagnosis and treatment of Parkinson's disease according to claim 1, characterized in that, Including the following: (1) To a medium containing HV and RVG 29 HATU and DIPEA were added to an ultrapure aqueous solution of PEG2k-NH2, and the mixture was stirred and subjected to a condensation reaction at room temperature for 3 hours to form RVG-linked PEG2k-NH2. 29 The compound HV-PEG-RVG 29 This step is crucial for ensuring the subsequent targeting of biological activity; (2) Dialysis bags with a molecular weight cutoff of 1000 kD were used to analyze the reaction product HV-PEG-RVG. 29 Dialysis was performed for 24 hours to remove unreacted small molecules and byproducts, yielding a product containing RVG. 29 HV probe.

3. A nanoparticle HVC NPs for the diagnosis and treatment of Parkinson's disease, characterized in that, The nanoparticle HVC NPs structure includes a probe 2-(2-(6-((4-(4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)benzyl)oxy)-2,3-dihydro-1H-thioanthrayl-4-yl)vinyl)-1-(2-carboxyethyl)-3,3-dimethyl-3H-indole-1-onium inner salt (HV) responsive to H2O2 and viscosity, and RVG linked to it. 29- The modified portion of PEG-NH2 and the loaded drug curcumin; RVG 29 -PEG-NH2 is derived from the targeting peptide RVG 29 Composed of polyethylene glycol (PEG) and amino groups (-NH2), the nanoparticles enhance their stability and targeting in vivo by being linked to an HV probe. Through a self-assembly process, nanoparticles with a core-shell structure are formed, where the core mainly contains curcumin and the shell consists of an HV probe and RVG. 29 -PEG-NH2 composition; RVG 29 The modification enables nanoparticles to effectively cross the blood-brain barrier, allowing curcumin to be used to treat Parkinson's disease.

4. The application of the HVC NPs nanoparticles as described in claim 3, characterized in that, HVC NP nanoparticles are used to prepare nanomedicines that can be dually detected for H2O2 and viscosity.

5. The application of the HVC NPs nanoparticles as described in claim 3, characterized in that, HVC NP nanoparticles are used to prepare nanomedicines for the detection of recovery of damage in a Parkinson's disease cell model at the cellular level.

6. The application of the HVC NPs nanoparticles as described in claim 3, characterized in that, HVC NP nanoparticles are used to prepare nanomedicines for the treatment of Parkinson's disease.

Images