A kind of nanoparticle based on polyphenol compound modification and its preparation and application

Abstract

The application relates to a kind of nanoparticles based on polyphenol compound modification and its preparation and application.The nanoparticles are mainly Cu 2‑x Se (0<=x<=1) nanoparticles, the surface is modified using polyphenol compounds, and nanoparticles with antioxidant protection are obtained by reasonable use of different surface modifiers.The preparation method can obtain nanoparticles with adjustable size, good water dispersibility, good biocompatibility and strong antioxidant effect.The application also discloses the application of nanoparticles after different modifications as new antioxidants, effective removal of reactive oxygen species (ROS), protection of neuronal cell damage and improvement of ischemia-reperfusion injury.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to a nanoparticle based on polyphenolic compound modification and its preparation and application. Background Technology

[0002] Neurodegenerative diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), and amyotrophic lateral sclerosis (ALS) have seriously threatened the health of the elderly. Among them, PD has become the second most common neurodegenerative disease in the world. Although there are various treatment methods, including drug therapy and surgical treatment, the complex and diverse pathogenesis of PD makes it difficult to cure it fundamentally. With the increasing aging of the population, many factors such as gene mutation and protein imbalance will accelerate the occurrence and development of PD, making the development of safe and reliable neuroprotective drugs particularly important. Currently, drugs widely used for neuroprotection include calcium ion channel blockers, glutamate receptor antagonists, free radical scavengers, and cell membrane stabilizers. However, these small molecule drugs are generally limited by the following problems: (1) poor water solubility, which greatly reduces their bioavailability; (2) easy degradation and metabolism, resulting in short circulation time in the body; (3) single protective effect, which cannot achieve multi-faceted and multi-angle neuroprotective effects.

[0003] Compared to small-molecule neuroprotective agents with well-defined structures and fixed properties, nanoparticles possess numerous advantages, including a larger specific surface area, modifiability, and therapeutic potential, and have been widely applied in preclinical research for various major diseases. By carefully designing and controlling the size, structure, and composition of nanomaterials, their physical and chemical properties can be effectively improved. Furthermore, surface functionalization of nanoparticles using various surface modifiers is a crucial strategy, not only enhancing their neuroprotective effects but also improving their dispersibility, biocompatibility, and targeting capabilities. Therefore, surface modification of nanoparticles is an important strategy for constructing nanomedicines with good water solubility, high safety, and multiple neuroprotective effects.

[0004] To construct such neuroprotective agents, patent application CN202210835211 discloses a hydrogel material for spinal cord injury protection and repair, synthesized from N-acryloylglycine, methacrylated gelatin, nanoclay, tannic acid, and small extracellular vesicles. Patent application CN202210109925 discloses a nano-drug delivery system developed using endogenous carrier mouse hippocampal cell exosomes to encapsulate the small molecule drug adenosine, demonstrating a protective effect against cognitive impairment caused by cerebral ischemia in mice. Patent application CN202111585161 discloses a Sargassum fusiforme polysaccharide nano-selenium and its application, which exhibits certain neuroprotective effects in a 6-hydroxydopamine-induced Parkinson's disease rat model; this nano-selenium is prepared using Sargassum fusiforme polysaccharide as a template. Patent application CN202110355826 discloses a method for preparing and applying self-assembled nanoparticles of diosgenin derivatives and DHA. The prepared nanoparticles can be used to protect microglia from LPS-induced inflammatory responses. Patent application CN202210411440 discloses phenolic compounds from hawthorn fruit, their preparation methods, and applications. It discloses that phenolic compounds prepared from hawthorn through a series of chemical methods can exert neuroprotective effects against Parkinson's disease. None of these patents involve modifying nanoparticles with antioxidant polyphenol molecules to construct nanoparticles that protect neurons and improve kidney damage. Summary of the Invention

[0005] To address the aforementioned technical problems, this invention provides nanoparticles modified with polyphenolic compounds, their preparation, and applications. This invention effectively enhances antioxidant capacity and increases bioavailability through the strong coordination between copper ions on the nanoparticle surface and polyphenols and flavonoids, while simultaneously providing essential trace elements copper and selenium for maintaining health. The nanoparticles are prepared using low-cost raw materials, have a simple construction method, exhibit strong antioxidant and neuroprotective effects, and demonstrate high biosafety, significantly overcoming challenges such as limited functionality, complex preparation, and difficulty in achieving integrated diagnostic and therapeutic applications.

[0006] This invention is achieved through the following technical solution:

[0007] The first objective of this invention is to provide nanoparticles modified with polyphenolic compounds, said nanoparticles comprising Cu 2-x Se nanoparticles and their surface modifiers, wherein the surface modifiers are polyphenolic compounds; wherein 0 ≤ x ≤ 1; the surface modifiers interact with Cu through chelation. 2-x Se nanoparticles are linked together.

[0008] In one embodiment of the present invention, the Cu 2-xThe particle size of Se nanoparticles ranges from 1 nm to 100 nm. Furthermore, considering that smaller nanoparticle sizes result in a larger specific surface area, which can provide more surface copper ions, nanoparticles with a size of less than 20 nm are preferred.

[0009] In one embodiment of the present invention, the polyphenolic compound is selected from flavonoid compounds, which are selected from one or more of curcumin, bilobetin, icariin, kaempferol, hesperetin, catechin, and anthocyanidin.

[0010] In one embodiment of the present invention, the Cu 2-x The mass ratio of Se nanoparticles to the surface modifier is 1:1 to 1:50.

[0011] In one embodiment of the present invention, the Cu 2-x Se nanoparticles are modified with stabilizers, including water-soluble thiol compounds and / or biocompatible molecules.

[0012] In one embodiment of the present invention, the water-soluble thiol compound includes mono- or poly-thiol small molecule organic compounds, including one or more of mercaptoethanol, mercaptopropanol, mercaptoacetic acid, mercaptopropionic acid, mercaptosuccinic acid, 2,3-dimercaptosuccinic acid, and mercaptoethylamine; the water-soluble thiol compound also includes a polymer polymer modified with mono- or poly-thiol, the polymer polymer including one or more of polyacrylic acid, polymethacrylic acid, and polyvinyl alcohol.

[0013] In one embodiment of the present invention, the biocompatible molecule includes natural polymers and artificial polymers. The natural polymers include one of dextran and its derivatives, chitosan and its derivatives, bovine serum albumin, and human serum albumin. The artificial polymers include one or more of the following: hydroxyl, carboxyl, amino, thiol, aldehyde, ester polyethylene glycol, homo-terminated bifunctional telechelic polyethylene glycol, hetero-terminated bifunctional telechelic polyethylene glycol, polyethylene glycol-polyacrylic acid copolymer, polyethylene glycol-polymethacrylic acid copolymer, polyethylene glycol-polyvinylamine copolymer, polyethylene glycol-polylactic acid copolymer, polyvinylpyrrolidone, polyethyleneimine, polyacrylic acid, and polyvinyl alcohol. The molecular weight of the polyethylene glycol is 200 to 20,000.

[0014] The second objective of this invention is to provide a method for preparing nanoparticles, which utilizes a one-pot method to prepare Cu nanoparticles of different sizes. 2-x Se nanoparticles, while utilizing Cu 2-xThe abundant divalent copper ions on the surface of Se nanoparticles can chelate with polyphenolic compounds, thus constructing nanoparticles with antioxidant properties. The specific steps include: adding a polyphenolic compound solution dropwise to Cu at a certain rate. 2-x In a Se nanoparticle solution, stirring accelerates the chelation process, ultimately yielding Cu modified with polyphenolic compounds. 2-x Se nanoparticles.

[0015] In one embodiment of the invention, the dripping rate is 1-5 drops per second. Specifically, it is 1 drop / s, 2 drops / s, 3 drops / s, 4 drops / s, 5 drops / s, etc., or any value between these values.

[0016] In one embodiment of the present invention, the solution obtained after stirring is further subjected to dialysis, ultrafiltration and purification to remove excess polyphenols and flavonoids, and finally obtain nanoparticles with antioxidant properties.

[0017] In one embodiment of the present invention, the stirring conditions are: stirring temperature of 0℃-30℃ and stirring time of 10min-12h.

[0018] In one embodiment of the present invention, the solvent for the polyphenol and flavonoid compound solution is one or more selected from ethanol, acetone, dimethyl sulfoxide (DMSO), and dimethylformamide (DMF). More preferably, dimethyl sulfoxide (DMSO) is used.

[0019] In one embodiment of the present invention, the concentration of the polyphenol compound solution is 1 mg / mL to 10 mg / mL.

[0020] In one embodiment of the present invention, the Cu 2-x The solvent for the Se nanoparticle solution is water and / or DMSO.

[0021] In one embodiment of the present invention, the Cu 2-x The concentration of the Se nanoparticle solution was 0.1 mg / mL to 1 mg / mL.

[0022] In one embodiment of the present invention, the polyphenolic compound and the Cu 2-x The molar ratio of Se nanoparticles is 1:1 to 1:10.

[0023] Further, the preparation method: Cu 2-xSe nanoparticles are dispersed in an aqueous solution, with the copper concentration maintained at 0.1 mg / mL-1 mg / mL. Then, an equal volume of organic solvent (such as DMSO, DMF, etc.) is added, followed by a solution of polyphenols and flavonoids. The mixture is stirred at a set temperature for a certain period of time. After stirring, nanoparticles with different surface modifications can be obtained through steps such as dialysis, centrifugation, and ultrafiltration.

[0024] A third objective of this invention is to provide the application of the aforementioned nanoparticles in antioxidants.

[0025] A fourth objective of this invention is to provide the application of the obtained nanoparticles in the preparation of neuroprotective agents.

[0026] The fifth objective of this invention is to provide the application of the obtained nanoparticles, antioxidants, and neuroprotective agents in drugs for neurodegenerative diseases and drugs for ischemia-reperfusion.

[0027] In one embodiment of the present invention, the neurodegenerative disease is Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), and amyotrophic lateral sclerosis (ALS).

[0028] The technical solution of the present invention has the following advantages compared with the prior art:

[0029] The present invention contains Cu nanoparticles with antioxidant properties. 2-x Se nanoparticles and their surface modifiers are composed of simple components. Cu 2-x The abundant divalent copper ions on the surface of Se nanoparticles can chelate with polyphenols and flavonoids, which not only effectively reduces the surface reduction potential and thus improves the antioxidant properties of the surface modifiers, but also improves their water solubility and bioavailability.

[0030] In addition to curcumin, ginkgol, and icariin, which are described in the examples, the polyphenols and flavonoids of this invention, including kaempferol, hesperidin, catechin, and anthocyanins, all belong to the polyphenol class. The phenolic hydroxyl groups on polyphenols have strong reducing properties; more phenolic hydroxyl groups mean more oxidizable sites, thus increasing reducing power. Furthermore, the redox potential is lowered after copper ions chelate with phenolic hydroxyl groups, significantly enhancing the reducing power of pure polyphenols.

[0031] The present invention provides nanoparticles with antioxidant protection that have adjustable size. The preferred ultra-small nanoparticles have a larger specific surface area than large nanoparticles, allowing for the modification of more ligands and making it easier to cross the blood-brain barrier, thus providing a basis for feasibility and safety in in vivo experiments.

[0032] The nanoparticles of this invention have superior antioxidant properties, and can achieve strong antioxidant capacity even at low concentrations. They not only provide protection, but also reduce the toxic side effects caused by large doses.

[0033] The nanoparticles of this invention have a long blood circulation time and exhibit MPP in vitro. + In induced Parkinson's disease models, it exhibits good protective effects, effectively reducing mitochondrial ROS, protecting mitochondria, maintaining intracellular redox balance, and reducing neuronal apoptosis, demonstrating high potential application value in in vivo treatment of Parkinson's disease.

[0034] The nanoparticles of this invention exhibit a good protective effect in a mouse model of renal ischemia-reperfusion injury, effectively scavenging ROS and demonstrating high potential application value.

[0035] The above description is merely an overview of the technical solution of the present invention. In order to better understand the technical means of the present invention and to implement it in accordance with the contents of the specification, the preferred embodiments of the present invention are described in detail below with reference to the accompanying drawings. Attached Figure Description

[0036] To make the content of this invention easier to understand, the invention will be further described in detail below with reference to specific embodiments and accompanying drawings, wherein...

[0037] Figure 1 This is a transmission electron microscope image of CSP nanoparticles in Example 1 of the present invention.

[0038] Figure 2 This is a transmission electron microscope image of BSA-CS nanoparticles in Example 1 of the present invention.

[0039] Figure 3 This is a transmission electron microscope image of PVP-CS nanoparticles in Example 1 of the present invention.

[0040] Figure 4 These are the CSC and CSP UV-Vis absorption spectra in Example 2 of this invention.

[0041] Figure 5 These are the UV-Vis absorption spectra of CSB and CSP in Example 3 of this invention.

[0042] Figure 6 These are the CSI and CSP UV-Vis absorption spectra in Example 4 of this invention.

[0043] Figure 7 The color is the aqueous solution color of CSP, CSC, CSB, and CSI in Examples 1-4 of this invention.

[0044] Figure 8The hydrated particle size of CSP, CSC, CSB, and CSI in Examples 1-4 of this invention is referred to as the hydrated particle size.

[0045] Figure 9 This is the ultraviolet-visible absorption spectrum of the BSA-CSC nanoparticles in Example 5 of this invention.

[0046] Figure 10 This is the ultraviolet-visible absorption spectrum of the PVP-CSC nanoparticles in Example 5 of this invention.

[0047] Figure 11 This represents the total antioxidant capacity of CSC nanoparticles at different concentrations in Example 6 of this invention.

[0048] Figure 12 This refers to the total antioxidant capacity of different nanoparticles at the same copper concentration in Example 6 of this invention.

[0049] Figure 13 This describes the effect of CSC nanoparticles on the survival rate of SH-SY5Y cells in Example 7 of this invention.

[0050] Figure 14 This describes the effect of CSC nanoparticles on ROS production in cellular mitochondria in Example 8 of this invention.

[0051] Figure 15 This describes the effect of CSC nanoparticles on neuronal cell mitochondria in Example 9 of this invention.

[0052] Figure 16 This describes the effect of CSC nanoparticles on the redox environment of neuronal cells in Example 10 of this invention.

[0053] Figure 17 This describes the effect of CSC nanoparticles on the apoptosis protein Cle-Caspase 3 in Example 11 of this invention.

[0054] Figure 18 This refers to the change in the antioxidant properties of CSC nanoparticles compared to curcumin alone in Example 12 of this invention.

[0055] Figure 19 This describes the effect of CSB nanoparticles on renal function in ischemia-reperfusion mice in Example 13 of this invention. Detailed Implementation

[0056] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, so that those skilled in the art can better understand and implement the present invention. However, the embodiments described are not intended to limit the present invention.

[0057] Preparation method of materials involved in this invention

[0058] 1. The Cu used in this invention 2-x The preparation method of Se nanoparticles (based on the method of Chinese Patent ZL201610213490.2) is as follows:

[0059] (1) Disperse one of selenium powder, sodium selenite, tellurium powder, and sodium tellurite in water at a concentration of 1 mmol / L to 100 mmol / L under inert gas protection.

[0060] (2) Add sodium borohydride powder to the dispersion system in step (1), the concentration of sodium borohydride is 30 mmol / L to 300 mmol / L, mix evenly, and reduce until the solution is colorless;

[0061] (3) Add the mixed aqueous solution of biocompatible molecules and metal cation precursors to a colorless low-valence chalcogenide anion solution, wherein the molar ratio of water-soluble thiol compound or biocompatible polymer to metal cation is 20:1 to 1:1, and stir the reaction at room temperature for 1 min to 3 h; the biocompatible molecules are bovine serum albumin, polyvinylpyrrolidone, etc.

[0062] 4) After the reaction is complete, the solution is separated by ultrafiltration multiple times to obtain the Cu used in this invention. 2-x Se nanoparticles.

[0063] Example 1:

[0064] This invention provides methods for preparing CSP, BSA-CS, and PVP-CS nanoparticles, the specific methods of which are as follows:

[0065] Using Cu of different sizes 2-x Se nanoparticles were used to obtain 1-3 nm polyvinylpyrrolidone-stabilized Cu. 2- x Se nanoparticles (CSP), 10-20nm bovine serum albumin-stabilized Cu 2-x Se nanoparticles (BSA-CS) and 70-100 nm polyvinylpyrrolidone-stabilized Cu 2-x Se nanoparticles (PVP-CS). Figure 1 , 2 Images 1, 2, and 3 are transmission electron microscopy (TEM) images of CSP, BSA-CS, and PVP-CS nanoparticles, respectively.

[0066] Example 2:

[0067] This embodiment provides a curcumin-modified nanoparticle CSC with neuroprotective effects, and the specific method is as follows:

[0068] The PVP-stabilized ultrasmall Cu from Example 1 2-xSe nanoparticle solution (CSP) was mixed with curcumin solution dissolved in DMSO at a molar ratio of 1:2 for nanoparticles to curcumin and a volume ratio of 1:1 for water to DMSO. The mixture was stirred at room temperature for 4 hours and then dialyzed for 24 hours in a dialysis bag with a molecular weight cutoff of 8k-1.4kDa to remove impurities. Finally, the mixture was centrifuged to remove the precipitate and concentrated by ultrafiltration to obtain CSC nanoparticles. Figure 4 The values ​​are CSC and CSP UV-Vis absorption spectra.

[0069] Example 3:

[0070] This embodiment provides a ginkgolic acid-modified CSB nanoparticle with neuroprotective effects, and the specific method is as follows:

[0071] The PVP-stabilized ultrasmall Cu from Example 1 2-x Se nanoparticle solution was mixed with ginkgol dissolved in DMSO at a molar ratio of 1:0.5, and the volume ratio of water to DMSO was maintained at 61:39. The mixture was stirred in an ice-water bath for 8 hours, and then dialyzed for 24 hours in a dialysis bag with a molecular weight cutoff of 8k-14kDa to remove impurities. Finally, the precipitate was removed by centrifugation, and the mixture was concentrated by ultrafiltration to obtain CSB nanoparticles. Figure 5 The images show the UV-Vis absorption spectra of CSB and CSP.

[0072] Example 4:

[0073] This embodiment provides icariin-modified CSI nanoparticles with neuroprotective effects, and the specific method is as follows:

[0074] The PVP-stabilized ultrasmall Cu from Example 1 2-x Se nanoparticle solution was mixed with icariin dissolved in DMSO, and PVP stabilized ultrasmall Cu 2-x The molar ratio of Se nanoparticles to icariin was 1:1, and the volume ratio of water to DMSO was maintained at 1:1. The mixture was stirred at room temperature for 4 hours, and then placed in a dialysis bag with a molecular weight cutoff of 8k-14kDa for dialysis for 24 hours to fully remove impurities. Finally, the precipitate was removed by centrifugation, and the mixture was concentrated by ultrafiltration to obtain CSI nanoparticles. Figure 6 The values ​​are CSI and CSP UV-Vis absorption spectra. Figure 7 , 8 The aqueous solutions of CSP, CSC, CSB, and CSI are respectively labeled with their colors and hydrated particle sizes.

[0075] Example 5:

[0076] This embodiment provides large-size Cu modified with curcumin. 2-x Se nanoparticles, the specific method is as follows:

[0077] The two large-sized Cu synthesized in Example 12-x Se nanoparticles were mixed with curcumin dissolved in DMSO, and Cu 2- x The molar ratio of Se nanoparticles to curcumin was 1:2, and the volume ratio of water to DMSO was maintained at 1:1. The mixture was stirred at room temperature for 4 hours, and an equal volume of DMSO was added and stirred until the solution was clear and transparent. After centrifugation at 20,000 rpm for 10 minutes, the supernatant was discarded, and the solution was reconstituted with water. This process was repeated twice to obtain curcumin-modified BSA-CS nanoparticles (BSA-CSC nanoparticles) and curcumin-modified PVP-CS nanoparticles (PVP-CSC nanoparticles), respectively. Figure 9 , 10 The UV-Vis absorption spectra of BSA-CSC nanoparticles and PVP-CSC nanoparticles are shown, respectively.

[0078] Example 6:

[0079] This embodiment provides the in vitro antioxidant capacity detection of the obtained nanoparticles.

[0080] The antioxidant properties of nanoparticles with concentrations of 6.25 μM, 12.5 μM, and 25 μM were determined using the total antioxidant capacity assay kit (ABTS rapid method), as well as the antioxidant properties of nanoparticles with different modifications at the same copper concentration. Figure 11 To demonstrate the antioxidant capacity of CSC nanoparticles at different concentrations, by Figure 11 It can be seen that the antioxidant capacity of CSC nanoparticles increases with increasing concentration. Figure 12 The total antioxidant capacity of nanoparticles with different modifications at the same copper concentration is calculated by... Figure 12 It was found that CSC nanoparticles exhibited the best antioxidant properties under the same copper concentration; therefore, CSC nanoparticles were selected for subsequent experimental testing. A one-way ANOVA was conducted using Graph Pad, with ***P<0.001 and ****P<0.0001.

[0081] Example 7:

[0082] This embodiment provides a detection method for how nanoparticles improve neuronal cell survival.

[0083] The neuroprotective effect of nanomaterials was detected using an enhanced CCK-8 assay kit: SH-SY5Y cells were seeded into 96-well plates, and then 25 μM CSC nanoparticles were incubated with SH-SY5Y cells for 2 h before being replaced with 3 mM MPP. + Continue co-incubation for 24 hours. After the incubation period, add 10 μL of CCK-8 solution to each well and incubate at 37°C in the dark for 1 hour. Then, measure the absorbance at 450 nm using a microplate reader. The experimental results are shown below. Figure 13As shown in the figure, CSC nanoparticles enhance the anti-MPP effect of SH-SY5Y cells. + Cell survival rate after damage. Graph Pad used one-way ANOVA, ****P<0.0001.

[0084] Example 8:

[0085] This embodiment provides a test of the ability of nanoparticles to scavenge free radicals in cellular mitochondria.

[0086] The antioxidant protective effect of nanoparticles was verified by detecting ROS in mitochondria using immunofluorescence staining: SH-SY5Y cells were seeded in an eight-well confocal dish, and 25 μM CSC nanoparticles were incubated with SH-SY5Y cells for 2 h. The cells were then replaced with 3 mM MPP. + Continue co-incubation for 24 hours. Then add 100 μL of mitochondrial ROS probe working solution to each well and incubate at 37°C in the dark for 1 hour. After washing twice with PBS, observe the fluorescence distribution and intensity in the cells using a laser confocal scanning microscope. Figure 14 The results show that CSC nanoparticles can effectively reduce MPP. + Damage caused by excessive ROS should be removed.

[0087] Example 9:

[0088] This embodiment provides the detection of the protective effect of nanoparticles on neuronal mitochondria.

[0089] The protective effect of nanoparticles on intracellular mitochondria of neurons was detected by immunofluorescence staining: SH-SY5Y cells were seeded in an eight-well confocal dish, and 25 μM CSC nanoparticles were pre-incubated with SH-SY5Y cells for 2 h, followed by replacement with 3 mM MPP. + Continue co-incubation for 24 hours. Then, add 100 μL of mitochondrial tracer probe working solution to each well and incubate at 37°C in the dark for 30 minutes. Afterward, wash twice with PBS. Observe the fluorescence distribution and intensity in the cells using a laser confocal scanning microscope. The experimental results are shown below. Figure 15 . Figure 15 The results show that CSC nanoparticles can effectively reduce mitochondrial damage in neuronal cells.

[0090] Example 10:

[0091] This embodiment provides a method for detecting the impact of nanoparticles on the intracellular redox environment of neuronal cells.

[0092] The effects of nanoparticles on intracellular redox activity in neurons were detected using a kit for detecting reduced glutathione (GSH) and oxidized glutathione (GSSG). The contents of total glutathione and reduced glutathione were measured sequentially according to the kit instructions; the difference between the two values ​​represented the oxidized glutathione content.

[0093] Finally, the GSH / GSSG ratio was obtained. The experimental group was divided into a control group (Ctrl) and an MPP group. + Group, CSC+MPP +

[0094] Group, experimental results are shown in Figure 16 . Figure 16 The results showed that CSC nanoparticles can protect the redox environment of neurons. A one-way ANOVA was performed on Graph Pad; *P<0.05, **P<0.01, and no significant difference was found in ns.

[0095] Example 11:

[0096] This embodiment provides the detection of the effect of nanoparticles on neuronal cell apoptosis.

[0097] The neuroprotective effect of nanoparticles was verified by detecting the activation expression of Cleaved-Caspase 3 using immunofluorescence: SH-SY5Y cells were seeded onto glass slides in 6-well plates, and then 25 μM CSCs were added.

[0098] Nanoparticles were incubated with SH-SY5Y cells for 2 hours, and then replaced with 3mM MPP. + Afterwards, continue co-incubation for 24 hours. After the culture is completed, perform the following steps: fixation, membrane rupture, blocking, incubation with primary antibody, binding with secondary antibody, nucleoside staining, and mounting.

[0099] Finally, the expression of Cleaved-Caspase 3 in cells was observed using laser confocal scanning microscopy. The experimental results are shown in [Figure number missing]. Figure 17 . Figure 17 The results show that CSC nanoparticles effectively reduce MPP. + Apoptosis-inducing proteins

[0100] Expression of Cleaved-Caspase 3.

[0101] Example 12:

[0102] This embodiment provides a comparative analysis of the ability of nanoparticles and curcumin alone to scavenge free radicals in cellular mitochondria.

[0103] The antioxidant protective effect of nanoparticles was verified by detecting ROS in mitochondria using immunofluorescence staining: SH-SY5Y cells were seeded into an eight-well confocal dish, and 25 μM CSC nanoparticles and 25 μM...

[0104] Curcumin alone was incubated with SH-SY5Y cells for 2 hours, and then 3mM MPP was added. + Continue co-incubation for 245 h. Then add 100 μL of mitochondrial ROS probe working solution to each well and incubate at 37°C in the dark for 1 h. Afterward, wash twice with PBS and finally observe the fluorescence distribution and intensity in the cells using a laser confocal scanning microscope. Figure 18 The results showed that CSC nanoparticles significantly enhanced the ability to scavenge excess mitochondrial ROS compared to curcumin alone.

[0105] Example 13:

[0106] This embodiment demonstrates the therapeutic effect of CSB nanoparticles on renal ischemia-reperfusion injury (I / R) in mice.

[0107] The effect of CSB nanoparticles on renal function in a renal I / R mouse model was observed. C57BL / 6 mice were randomly divided into three groups: Sham group, IRI+PBS group, and IRI+CSB group (n=3). Mice were fasted for 8-12 hours preoperatively. Mice were anesthetized by intraperitoneal injection of 2.5% sodium pentobarbital. The renal pedicle was exposed through a dorsal incision, and the pedicle was quickly clamped with a non-invasive vascular clamp. The renal ischemia was maintained for 30 minutes before the clamp was released to restore blood perfusion. The surgical incision was then sutured in layers. The sham-operated group did not have the renal pedicle clamped; all other procedures were the same. The model was successfully established. Postoperatively, mice were fed routinely, and medication was administered: the CSB nanoparticle group received 100 μg / mL (200 μL) via tail vein administration at 2 hours and 24 hours, respectively. The model group received the same PBS. Weight changes were recorded throughout the process, and blood was collected from the eyeballs for renal function testing. Figure 19 This indicates that CSB nanoparticles can effectively improve kidney injury caused by renal ischemia-reperfusion in mice.

[0108] In summary, this invention presents a method for preparing highly biocompatible antioxidant nanoparticles and their application in inhibiting neurotoxicity and improving renal function. The method involves a simple process of stirring, mixing, dialysis, ultrafiltration, and centrifugation to obtain antioxidant nanoparticles. This preparation method is mild, inexpensive, rapid, and efficient, producing nanoparticles with uniform size, good water solubility, and good biocompatibility. Furthermore, due to the presence of Cu... 2-x The excellent photothermal conversion properties of Se nanoparticles can also be used for potential in vivo photoacoustic imaging, enabling integrated targeted therapy.

[0109] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the technical principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A nanoparticle based on modification of polyphenolic compounds, characterized by, The nanoparticles include Cu 2-x Se nanoparticles and their surface modifiers, wherein the surface modifiers are polyphenolic compounds; wherein 0 ≤ x ≤ 1; the surface modifiers interact with Cu through chelation. 2-x Se nanoparticles are connected; the polyphenolic compound is selected from flavonoids; the flavonoid compound is selected from one or more of curcumin, ginkgol, icariin, kaempferol, hesperidin, catechin and anthocyanin; the Cu 2-x Se nanoparticles are modified with a stabilizer, namely polyvinylpyrrolidone; the Cu... 2-x The Se nanoparticles have a particle size of 1 nm-100 nm; the Cu 2-x The molar ratio of Se nanoparticles to the surface modifier is 1:0.5-1:

2.

2. A method of producing nanoparticles as claimed in claim 1, characterized in that, Comprising the steps of: The polyphenolic compound solution is added to Cu 2-x The polyphenolic compound-modified Cu 2-x Se nanoparticles are obtained by stirring in the Se nanoparticle solution.

3. The preparation method according to claim 2, characterized in that, The solution obtained after stirring is also subjected to dialysis, ultrafiltration and purification.

4. Use of the polyphenol compound-based modified nanoparticle of claim 1 in the preparation of a neuroprotective preparation.

5. The polyphenol-modified nanoparticles according to claim 1, and the polyphenol-modified Cu nanoparticles obtained by the preparation method according to any one of claims 2-3. 2-x The application of Se nanoparticles and the neuroprotective agent described in claim 4 in the preparation of drugs for neurodegenerative diseases and drugs for ischemia-reperfusion.

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