Preparation method and application of gold nanometer "garland" with ability of treating acute liver injury

Abstract

The application belongs to the technical field of biological medicine, and discloses a preparation method and application of gold nanometer 'crown' with the ability of treating acute liver injury. Specifically disclosed is a gold nanometer crown, wherein hydroxylamine is loaded in the gold nanometer crown. The application stably loads hydroxylamine in the gold nanometer crown, improves the biocompatibility of hydroxylamine, enhances the treatment effect of hydroxylamine through the characteristics of the gold nanometer crown, and integrates targeting and slow release, thereby showing potential biological efficiency. The gold nanometer crown can specifically target liver cells, and can reduce systemic side effects related to conventional treatment. The targeting method can enhance the effect of the therapeutic agent carried by the nanoparticles.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a method for preparing and applying a gold nano "garland" capable of treating acute liver injury. Background Technology

[0002] Gold, a chemically very stable metallic element, exhibits unique biocompatibility, high electron density, and catalytic properties at the nanoscale. By controlling the morphology of gold nanoparticles, their photothermal properties and electric field strength can be tuned, making them widely applicable in fields such as cancer photothermal therapy, biosensors, and surface plasmon resonance enhanced spectroscopy (SERS). Hydroxylamine (NH₂OH·HCl) is a commonly used antioxidant and stabilizer, widely used in the production and reactions of various materials. However, hydroxylamine has extremely poor biocompatibility, limiting its application in vivo.

[0003] Therefore, this application is hereby submitted. Summary of the Invention

[0004] This invention aims to address at least one of the technical problems existing in the prior art. To this end, this invention proposes a gold nanorod "garland" (Au nanowreaths, AuNwrs) with therapeutic capabilities for acute liver injury. Through research, a method for stably loading hydroxylamine into the gold nanorods was proposed. This not only improves the biocompatibility of hydroxylamine but also enhances its therapeutic effect through the properties of the gold nanorods, thus combining targeted and sustained release, demonstrating potential bioefficacy. By stably loading hydroxylamine into the gold nanorods, new diagnostic and therapeutic approaches can be provided for the clinical treatment of liver injury and the blocking of the progression from acute liver injury to chronic liver fibrosis.

[0005] The first objective of this invention is to provide a gold nanogarland.

[0006] The second objective of this invention is to provide a method for preparing the gold nanogarland of the first aspect of this invention.

[0007] The third aspect of this invention aims to provide an application of the gold nanogarland of the first aspect of this invention.

[0008] The fourth aspect of this invention is to provide a drug.

[0009] The fifth aspect of this invention aims to provide a drug delivery system.

[0010] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0011] In a first aspect, the present invention provides a gold nanorod loaded with hydroxylamine.

[0012] In some embodiments of the present invention, the gold nanorods are obtained by the self-assembly of gold nanocorals via intermolecular hydrogen bonds of hydroxylamine.

[0013] In some embodiments of the present invention, the gold nanocoral is prepared by the following method: mixing a gold salt solution with a reducing agent and a gold seed solution, reacting to obtain the gold nanocoral.

[0014] In some embodiments of the present invention, the preparation process of the gold seed solution includes: mixing a reducing agent with a gold salt solution to obtain the gold seed solution.

[0015] In some embodiments of the present invention, the reducing agent is selected from at least one of citrate, ascorbic acid, sodium borohydride and hydroxylamine.

[0016] In some embodiments of the present invention, the gold salt is at least one of chloroauric acid and chloroaurate; preferably HAuCl4.

[0017] In some embodiments of the present invention, the reducing agent in the preparation of the gold seed solution includes citrate and sodium borohydride.

[0018] In some embodiments of the present invention, the reducing agent in the preparation process of the gold nanocoral includes citrate, sodium borohydride and hydroxylamine.

[0019] In some embodiments of the present invention, the preparation process of the gold seed solution includes: mixing NaBH4 with HAuCl4 and trisodium citrate solution, stirring until the solution turns pink, thus obtaining the gold seed solution.

[0020] In some preferred embodiments of the present invention, the concentration ratio of NaBH4, HAuCl4 and trisodium citrate is 1:1 to 2:10 to 12; preferably 1:1:12.

[0021] In some embodiments of the present invention, the gold nanocoral is prepared by the following method: HAuCl4 solution and trisodium citrate are mixed and heated at 30-40°C until the solution turns red or orange, the solution is collected and cooled to room temperature; hydroxylamine (NH2OH·HCl) solution is added, and after mixing, gold seed solution is added and reacted (stirred at 60-90°C for 20-200 min) to obtain the gold nanocoral.

[0022] By adjusting the reaction conditions during the preparation of gold nanocorals, gold nanocorals of different diameters can be prepared. The diameter of the gold nanocorals continuously increases with increasing stirring time.

[0023] In some preferred embodiments of the present invention, the reaction conditions are stirring at 65–85°C for 30–150 min.

[0024] This invention provides a specially designed gold nanorod for targeted hepatocyte therapy, aiming to offer a new approach for the clinical treatment of acute liver injury and the prevention of liver fibrosis. This novel nanorod possesses considerable biocompatibility and antioxidant capacity. Furthermore, by altering the surface of the gold nanoparticles, novel nanorods with different particle sizes and carrying different drugs can be customized to modulate inflammatory responses in different organs, which is crucial in acute organ injury. These nanoparticles not only possess inherent anti-inflammatory properties but also directly deliver antioxidants to hepatocytes, thereby more effectively reducing oxidative stress and avoiding excessive side effects compared to traditional systemic antioxidant therapy.

[0025] The gold nanorods provided by this invention have good biocompatibility and a longer metabolic cycle in vivo than most clinical drugs. They can effectively correct inflammation and oxidative stress products that occur in acute liver injury, reduce tissue damage, and treat acute liver injury and block the process of inflammatory transformation into fibrosis.

[0026] A second aspect of the present invention provides a method for preparing the gold nanorods of the first aspect of the present invention, comprising the following steps: mixing a gold salt solution with citrate and hydroxylamine, mixing with gold nanocoral under stirring conditions, and reacting to obtain gold nanorods.

[0027] In some embodiments of the present invention, the preparation method includes mixing HAuCl4 solution and trisodium citrate at 30-40°C until the solution turns red or orange, collecting the solution and cooling it to room temperature; adding hydroxylamine (NH2OH·HCl) solution, mixing it with gold nanocoral under stirring conditions, and reacting to obtain gold nanogarlands.

[0028] In some embodiments of the present invention, the reaction time is 20 to 150 minutes.

[0029] In some preferred embodiments of the present invention, the reaction time is 30 to 150 minutes.

[0030] In some embodiments of the present invention, the final concentration of the hydroxylamine in the reaction system is 0.01 to 0.1 M.

[0031] In some preferred embodiments of the present invention, the final concentration of the hydroxylamine in the reaction system is 0.01 to 0.08 M.

[0032] In some preferred embodiments of the present invention, the final concentration of the hydroxylamine in the reaction system is 0.01 to 0.06 M.

[0033] The diameter of gold nanorods can be controlled by adjusting the concentration of hydroxylamine in the reaction system.

[0034] Hydrogen chloroaurate (HAuCl4) provides gold ions for reduction by NaBH4. During reduction, these ions form gold atoms, which aggregate into nanoparticles, serving as the primary material for nanocoral structures. Hydroxylamine (NH2OH-HCl) can be used both to reduce and generate gold nanorods and to control the growth kinetics of these structures. It provides a different reduction potential than NaBH4, allowing manipulation of the shape and surface properties of the gold nanoparticles. This method utilizes the unique properties of each reagent to control the nucleation and growth stages of gold nanoparticles, forming complex nanorod structures with potentially high-impact applications.

[0035] The method for preparing gold nanorods provided by this invention is simple to operate, and the entire process requires no reagents other than sodium borohydride, hydrogen tetrachloroaurate, trisodium citrate, and hydroxylamine. This method offers high reproducibility, low cost, and produces gold nanorods with a hollow structure, uniform size, good dispersibility, and adjustable size and wall thickness.

[0036] A third aspect of the present invention provides the application of the gold nanorod of the first aspect of the present invention in at least one of (1) to (5):

[0037] (1) Prepare products for the treatment and / or prevention of liver diseases;

[0038] (2) Drug delivery;

[0039] (3) Preparation of a drug delivery system;

[0040] (4) Preparation of drug carriers;

[0041] (5) Prepare products for monitoring the distribution and / or location of drugs.

[0042] In some embodiments of the present invention, the liver disease described in (1) includes acute liver injury.

[0043] In some preferred embodiments of the present invention, the liver disease described in (1) includes carbon tetrachloride-induced acute liver injury.

[0044] The gold nanorods provided by this invention, characterized by their biocompatibility and non-toxicity at specific concentrations, offer a safer alternative to some potentially serious side effects for patients with impaired liver function. Furthermore, the unique properties of gold allow for real-time in vivo imaging and monitoring of these nanoparticles using techniques such as CT scans or MRI, enabling precise tracking of treatment distribution and adjustments to treatment strategies based on real-time data. The versatility of the gold nanorods also allows for their combination with other therapeutic modalities, such as drug therapy or gene therapy, to create synergistic effects and enhance overall treatment efficacy. This versatile and adaptable nanoparticle system represents a significant advancement in the treatment of acute organ injuries, improving both the effectiveness and safety of treatment modalities.

[0045] In a fourth aspect, the present invention provides a medicament comprising the gold nanogarland of the first aspect of the present invention.

[0046] In some embodiments of the present invention, the drug may also include pharmaceutically acceptable excipients.

[0047] In some embodiments of the present invention, the pharmaceutically acceptable excipients include at least one selected from fillers, disintegrants, diluents, lubricants, binders, humectants, flavoring agents, suspending agents, solvents, sustained-release agents, emulsifiers, absorption enhancers, surfactants, preservatives, pigments, fragrances, and solvents.

[0048] In some embodiments of the present invention, the drug may further include a drug for treating acute liver injury.

[0049] A fifth aspect of the present invention provides a drug delivery system comprising a gold nanorod of the first aspect of the present invention, and a drug loaded on the gold nanorod.

[0050] In some embodiments of the present invention, the drug includes a drug for treating acute organ injury.

[0051] The beneficial effects of this invention are:

[0052] This invention stably loads hydroxylamine into gold nanorods, which not only improves the biocompatibility of hydroxylamine but also enhances its therapeutic effect through the properties of the gold nanorods, thus combining targeted and sustained release and demonstrating potential bioefficacy. These gold nanorods can specifically target hepatocytes, potentially reducing systemic side effects associated with conventional treatments. This targeting approach can enhance the efficacy of therapeutic agents carried by nanoparticles.

[0053] Furthermore, the nanogarland provided by this invention has the following characteristics:

[0054] Anti-inflammatory properties: Gold nanoparticles have been documented to possess anti-inflammatory properties. By modifying their surface, gold nanocoral can be tailored to modulate inflammatory responses in the liver, which is crucial in acute liver injury.

[0055] Antioxidant properties: Oxidative stress plays a crucial role in the progression of liver injury. Gold nanorods can directly deliver and release antioxidants to damaged hepatocytes, thereby effectively alleviating oxidative stress (reducing ALT and AST levels).

[0056] Biocompatibility and safety: Gold atoms are biocompatible and non-toxic at certain concentrations. Gold nanorods loaded with hydroxylamine can greatly improve the biocompatibility of hydroxylamine.

[0057] Real-time monitoring: The unique properties of gold atoms allow for the imaging and tracking of these nanoparticles in vivo using techniques such as CT scans or MRI. This helps monitor the distribution and localization of treatments, providing real-time feedback for adjusting treatment strategies.

[0058] Combination therapy: Gold nanorods can be used in combination with other treatments (such as drug therapy or gene therapy) to create a synergistic effect and enhance the overall treatment outcome. Attached Figure Description

[0059] Figure 1 This is a flowchart of the preparation process for AuNCls and AuNwrs.

[0060] Figure 2 Characterization results of gold seeds (AuNDs); where a is a TEM electron microscope image; b is a particle size distribution histogram; c is a SAED image;

[0061] Figure 3 The images show TEM electron micrographs of AuNCls at different reaction times when the reactant ratio is 1; where a is the TEM electron micrograph at a reaction time of 30 min; b is the TEM electron micrograph at a reaction time of 60 min; c is the TEM electron micrograph at a reaction time of 90 min; d is the TEM electron micrograph at a reaction time of 120 min; e is the TEM electron micrograph at a reaction time of 150 min; f to j are the high-resolution TEM electron micrographs corresponding to a to e; k is the XRD pattern of AuNCls of different sizes.

[0062] Figure 4The images show schematic diagrams of AuNwrs synthesis and TEM images of AuNwrs at different sizes; where a is a schematic diagram of AuNwrs synthesis; b is a TEM image of 58 nm AuNwrs; c is a TEM image of 79 nm AuNwrs; d and e are TEM images of 92 nm AuNwrs; f is a TEM image of 148 nm AuNwrs; g is a TEM image of 274 nm AuNwrs; and h is a TEM image of 360 nm AuNwrs.

[0063] Figure 5 Fluorescence imaging of ex vivo organs of mice 12, 24, 48 and 72 h after intravenous injection of Cy7-modified 100nm-AuNwrs and 400nm-AuNwrs materials. In the figure, from left to right and from top to bottom, they represent heart, liver, spleen, lung, kidney, thymus, small intestine, muscle and brain.

[0064] Figure 6 HE staining results of heart, liver, spleen and lung tissues of mice 7 days after gold nanorods were injected into mice via the tail vein.

[0065] Figure 7 The figures show the changes in liver function-related indicators in a carbon tetrachloride-induced liver injury model mouse before and after treatment with silymarin and gold nanorods. In the figures, A represents the survival rate of mice in different treatment groups; B represents the NO content in the liver of mice in different treatment groups after 7 days of treatment; C represents the IL-6 content in the liver of mice in different treatment groups after 7 days of treatment; D represents the ALT content in the liver of mice in different treatment groups after 7 days of treatment; E represents the AST content in the liver of mice in different treatment groups after 7 days of treatment; and F represents the bilirubin content in the liver of mice in different treatment groups after 7 days of treatment. In the figures, * and # indicate P < 0.05.

[0066] Figure 8 Comparison of the structure of liver tissue pathological sections stained with HE in a carbon tetrachloride-induced liver injury model mouse before and after treatment with silymarin and gold nanorods under a 200x microscope. Detailed Implementation

[0067] The present invention will be further described in detail below through specific embodiments.

[0068] It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention.

[0069] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0070] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0071] Example 1

[0072] A method for preparing gold nanocoral includes the following steps:

[0073] (1) Preparation of gold seeds (AuNDs)

[0074] Prepare 20 mL of a solution containing 2.5 × 10⁻⁶ ppm in an Erlenmeyer flask. -4 M hydrogen tetrachloroaurate (HAuCl4) and 2.5 × 10 -4 Add 0.6 mL of freshly prepared 0.1 M sodium borohydride (NaBH4) solution (prepared after being frozen) to an aqueous solution of M trisodium citrate (Na3cit), and stir continuously until the solution turns pink, indicating the formation of gold seeds, thus obtaining gold seeds (referred to as seed solution).

[0075] (2) Preparation of gold nanocoral (AuNCls)

[0076] First, prepare 2.5 × 10⁻⁶ ppm in an Erlenmeyer flask. -4 200 mL of M HAuCl4 solution was prepared, and 6 g of Na3cit was added (final concentration 0.08 M). The reaction was carried out at 35 °C until the solution turned orange. The collected solution was cooled to room temperature and recorded as the growth stock solution.

[0077] Next, prepare a 50mL conical flask and mix 7.5mL of the growth stock solution with 0.05mL of freshly prepared 0.1M NH2OH·HCl solution. Add 3.5mL of the seed solution obtained in step (1) and stir at 85℃ for 30min. The solution turns wine red, and AuNCls is obtained.

[0078] Analysis of the TEM electron microscope images showed that the diameter of the AuNCls was 23 nm.

[0079] Example 2

[0080] A method for preparing gold nanocoral differs from Example 1 only in that the reaction conditions during the preparation of AuNCls are changed from stirring at 85°C for 30 min to stirring at 85°C for 150 min.

[0081] The diameter of the AuNCls was determined to be 48 nm.

[0082] Example 3

[0083] A method for preparing gold nanocoral differs from Example 1 only in that the reaction conditions during the preparation of AuNCls are changed from stirring at 85°C for 30 min to stirring at 85°C for 60 min.

[0084] The diameter of the AuNCls was measured to be 27 nm.

[0085] Example 4

[0086] A method for preparing gold nanocoral differs from Example 1 only in that the reaction conditions during the preparation of AuNCls are changed from stirring at 85°C for 30 min to stirring at 85°C for 90 min.

[0087] The diameter of the AuNCls was determined to be 35 nm.

[0088] Example 5

[0089] A method for preparing gold nanocoral differs from Example 1 only in that the reaction conditions during the preparation of AuNCls are changed from stirring at 85°C for 30 min to stirring at 85°C for 120 min.

[0090] The diameter of the AuNCls was determined to be 43 nm.

[0091] Example 6

[0092] An antioxidant gold nano "garland" (AuNwrs) is formed by the self-assembly of gold nano corals prepared in Example 1 through intermolecular hydrogen bonds between NH2OH·HCl.

[0093] The preparation method of the above-mentioned gold nano "garlands" (AuNwrs) includes the following steps: Take 9 mL of growth stock solution (prepared in an Erlenmeyer flask with a concentration of 2.5 × 10⁻⁶ ppm) -4 200 mL of M HAuCl4 solution was mixed with 6 g of Na3cit (final concentration 0.08 M). The reaction was carried out at 35 °C until the solution turned orange, and the collected solution was cooled to room temperature. This mixture was then combined with 0.45 mL of 0.1 M NH2OH·HCl solution, and 1.0 mL of AuNCls from Example 1 was added under vigorous stirring. After reacting at room temperature for 30 min, AuNwrs was obtained.

[0094] Analysis of high-resolution TEM electron microscopy images revealed that the diameter of this AuNwrs is 148 nm.

[0095] Example 7

[0096] An antioxidant gold nano "garland" (AuNwrs) differs from Example 6 only in that the gold nano coral was prepared in Example 2, and the amount of 0.1M NH2OH·HCl solution used in the preparation of AuNwrs is different (0.45 mL of 0.1M NH2OH·HCl solution was replaced with 10.8 mL of 0.1M NH2OH·HCl solution).

[0097] Analysis of high-resolution TEM electron microscopy images revealed that the diameter of AuNwrs is 360 nm.

[0098] Example 8

[0099] An antioxidant gold nano "garland" (AuNwrs) differs from Example 6 only in that the gold nano coral was prepared in Example 3.

[0100] Analysis of high-resolution TEM electron microscopy images revealed that the diameter of this AuNwrs is 125 nm.

[0101] Example 9

[0102] An antioxidant gold nano "garland" (AuNwrs) differs from Example 7 only in that the gold nano coral was prepared in Example 4.

[0103] Analysis of high-resolution TEM electron microscopy images revealed that the diameter of this AuNwrs is 150 nm.

[0104] Example 10

[0105] An antioxidant gold nano "garland" (AuNwrs) differs from Example 6 only in that the gold nano coral was prepared in Example 5, and the room temperature reaction time during the preparation of AuNwrs was 150 min.

[0106] Analysis of high-resolution TEM electron microscopy images revealed that the diameter of this AuNwrs is 250 nm.

[0107] Example 11

[0108] An antioxidant gold nano-garland (AuNwrs) differs from Example 10 only in that Cy7 dye was added during the preparation process. The gold nano-coral was prepared in Example 1. Specifically, 9 mL of the growth stock solution was mixed with 0.45 mL of 0.1 M NH2OH·HCl solution, and 0.5 mg of Cy7 dye and 1.0 mL of AuNCls from Example 1 were added under vigorous stirring. After reacting for 150 min, AuNwrs (denoted as Cy7-AuNwrs) were obtained.

[0109] Analysis of high-resolution TEM electron microscopy images revealed that the diameter of this AuNwrs is 148 nm.

[0110] Example 12

[0111] An antioxidant gold nano "garland" (AuNwrs) differs from Example 11 only in that the amount of 0.1M NH2OH·HCl solution used in the preparation of AuNwrs is 10.8 mL.

[0112] Analysis of high-resolution TEM electron microscopy images revealed that the diameter of this AuNwrs is 360 nm.

[0113] Effect Example

[0114] 1. Preparation and characterization of gold nanocorals and gold nanogarlands

[0115] Figure 1 The preparation process of AuNCls and AuNwrs is shown in the figure. AuNCls is prepared by a two-step bottom-up method. First, gold seeds (i.e., gold nanodots, AuNDs) are synthesized using NaBH4 as a reducing agent. Figure 2 As shown in a and b, the gold seed is a nanodot structure of approximately 7.5 nm. Figure 2 c is the selected area electron diffraction (SAED) pattern of AuNDs. As can be seen from the figure, the diffraction peak of Au(111) plane is the strongest. This crystal plane has a low energy and is the main site for subsequent secondary growth.

[0116] Subsequently, AuNDs were mixed with HAuCl4, Na3cit, and the reducing agent NH2OH·HCl. With increasing reaction time, the size and number of AuNDs coral branches increased, exhibiting a multi-branched structure. For example... Figure 3 As shown in Figures a to e, when the reaction time increases from 30 min to 150 min, the size of AuNCls increases from about 23 nm (Example 1) to about 48 nm (Example 2) with the reaction time. Figure 3 High-resolution TEM images in f-j show that AuNCls obtained at different growth times all exhibit lattice fringes with spacings of 0.235 nm and 0.203 nm, which is consistent with the lattice structures of the Au(111) and Au(100) crystal planes. This result indicates that the series of nanoparticles prepared by changing the reaction time have the same crystal orientation and lattice structure. Further XRD diffraction of AuNCls yielded the following results: Figure 3As shown in Figure k, AuNCls exhibits XRD diffraction peaks at 38.19°, 44.39°, 64.58°, 77.57°, and 81.72°, which are assigned to the Au(111), (200), (220), (111), and (420) planes, respectively. This result is consistent with the results shown by the lattice fringes above.

[0117] like Figure 4 As shown in b to d, with N H2 Increasing the OH ratio alters the reactant ratio by changing the amount of NH₂OH·HCl. Figure 4 As can be seen from steps e to f, the reaction system at this time will assemble AuNWrs structures with a size of approximately 148–360 nm using AuNCls (Example 2) as the basic unit. These results demonstrate that the size of AuNWrs can be controlled by changing the reaction time.

[0118] The above conclusions suggest that the driving force for the self-assembly of AuNWrs is the intermolecular hydrogen bonds formed between excess NH2OH·HCl. When the amount of NH2OH·HCl is insufficient and the amount of Na3cit is abundant, the electrostatic repulsion between the -COOH in Na3cit exists in the form of -COO- in solution, thus maintaining the AuNCls in the form of single particles in solution. As the amount of NH2OH·HCl gradually increases, intermolecular hydrogen bonds will form between them, thereby initiating the assembly of AuNCls. Excess hydroxylamine is also adsorbed into the gold nanorods and released as the gold nanorods degrade.

[0119] The properties of NH₂OH·HCl and Na₃cit on gold surfaces were further investigated using density functional theory (DFT). Calculations showed that the relative adsorption energy of Na₃cit on Au(100) was 3.59 kJ / mol, higher than its relative adsorption energy on the Au(111) crystal plane (3.52 kJ / mol). The adsorption of NH₂OH·HCl on Au(100) and Au(111) was exactly the opposite. Figure 3 These results indicate that NH2OH·HCl is more readily attached to the Au(111) surface, while Na3cit is more readily adsorbed onto the Au(100) surface, allowing both to coexist on the AuNCls surface. Intermolecular hydrogen bonds of Na3cit drive self-assembly, and the electrostatic repulsion between Na3cit molecules helps maintain the hollow structure inside AuNwrs.

[0120] 2. Kidney-targeting studies of AuNwrs

[0121] The prerequisite for nanocarriers to achieve efficient treatment is good organ targeting. Here, male C57BL / 6 mice were selected as experimental mice. Cy7-labeled 0.5 mg / kg AuNwrs (i.e., the AuNwrs in Examples 11 and 12, 100 nm and 400 nm respectively) were administered via tail vein injection. Mice were euthanized at different time points, and their hearts, livers, spleens, lungs, kidneys, thymus, small intestines, muscles, and brains were collected for ex vivo organ fluorescence imaging.

[0122] The results are as follows Figure 5 As shown, Figure 5 Fluorescence imaging of isolated organs from mice at 0.5, 1, 2, and 3 days after drug administration. Figure 5 It can be seen that 100nm-AuNwrs has a significant liver enrichment effect and can maintain liver targeting for up to 48 hours. 400nm-AuNwrs exhibits the classic liver-kidney metabolic pathway, that is, it is mainly concentrated in the liver before 12 hours, and can be enriched in the kidneys after 12 hours and remain there until it disappears after 72 hours.

[0123] Tissue samples from the heart, liver, spleen, and lungs of mice injected via the tail vein (100 nm of AuNwrs) were collected 7 days later and stained with hematoxylin and eosin (HE). The results are as follows: Figure 6 As shown.

[0124] The above results fully demonstrate that the controllable particle size of gold nanorods brings unlimited prospects for targeted therapy. The change in particle size allows it to easily select the desired target organ site, providing important support for the next step of disease treatment.

[0125] 3. Therapeutic effect of 100nm-AuNwrs on a mouse model of carbon tetrachloride-induced acute liver injury

[0126] To investigate the therapeutic effect of 100 nm diameter AuNwrs (Example 6) on a mouse model of carbon tetrachloride-induced acute liver injury, male C57BL / 6 mice were selected as experimental animals. A mouse model of liver disease simulating acute liver injury was established by intraperitoneal injection of carbon tetrachloride solution. The experimental mice were divided into four groups: sham operation group, model group (0.5% carbon tetrachloride intraperitoneal injection 12 mL / kg), model + Yangshen control group (silymarin 50 mg / mL), and model + experimental group (AuNwrs 0.5 mg / kg from Example 6). After a single intraperitoneal injection of 12 mL / kg body weight of 0.5% carbon tetrachloride, various therapeutic drugs were injected via tail vein according to the group, and this was continued for 7 days, every other day. At 7 days, blood and liver samples were collected from 5 euthanized mice in each group and compared at both the molecular level (using ELISA kits to determine the molecular levels of NO and IL-6, and using a Beckman blood biochemistry analyzer to detect the levels of ALT, AST, and bilirubin) and the tissue level (HE staining).

[0127] The results are as follows Figure 7 As shown, the mortality rate of mice in the model group during the 7-day liver injury experiment was significantly higher than that in the treatment group (i.e., the model + Yangshen control group and the model + experimental group) and the sham group. Figure 7 Among them, gold nanorods showed effective therapeutic effects on carbon tetrachloride-induced liver injury, and their NO, IL-6, ALT, AST, and bilirubin levels were similar to those of the positive control group. Figure 7 (Middle B~F). From Figure 8 HE staining results also showed that both the positive control group and the nano-garland administration group had a certain therapeutic effect on acute liver injury, and the degree of liver inflammatory cell infiltration was much lower than that of the liver tissue of the model group mice.

[0128] In summary, this invention provides a specially designed gold nanorods with specific tracing particle sizes and morphologies for targeted hepatocytes, aiming to offer a new approach for the clinical treatment of acute liver injury and the prevention of liver fibrosis. These novel nanorods possess considerable biocompatibility and antioxidant capacity. Furthermore, utilizing the passive targeting principle of different organ uptake of nanoparticles with varying particle sizes, by altering the surface of the gold nanoparticles, novel nanorods with different particle sizes and carrying different drugs can be customized to modulate inflammatory responses in different organs, which is crucial in acute organ injury. These nanoparticles not only possess inherent anti-inflammatory properties but can also directly deliver antioxidants to hepatocytes, thereby more effectively reducing oxidative stress and avoiding excessive side effects compared to traditional systemic antioxidant therapy.

[0129] The gold-based nanosystem used in this invention, called gold nanorods, is characterized by its biocompatibility and non-toxicity at specific concentrations, providing a safer alternative to some drugs that may have serious side effects for patients with impaired liver function. Furthermore, the unique properties of gold allow for real-time in vivo imaging and monitoring of these nanoparticles using techniques such as CT scans or MRI, enabling precise tracking of treatment distribution and adjustment of treatment strategies based on real-time data.

[0130] Furthermore, this invention leverages the versatility of gold nanorods, combining them with other therapeutic modalities (such as drug therapy or gene therapy) to create synergistic effects and enhance overall therapeutic efficacy. This versatile and adaptable nanoparticle system represents a significant advancement in targeted therapy for acute organ injury, improving both the effectiveness and safety of treatment approaches.

[0131] The embodiments of the present invention have been described in detail above with reference to the accompanying drawings. However, the present invention is not limited to the above embodiments, and various changes can be made within the scope of knowledge possessed by those skilled in the art without departing from the spirit of the present invention. Furthermore, the embodiments of the present invention and the features thereof can be combined with each other unless otherwise specified.

Claims

1. A gold nanogarland, characterized in that, The gold nanorods are loaded with hydroxylamine; the gold nanorods are obtained by the self-assembly of gold nanocorals through intermolecular hydrogen bonds of hydroxylamine; The gold nano coral is prepared by the following method: mixing gold salt solution with reducing agent and gold seed solution, stirring at 60-90℃ for 20-200 min to obtain gold nano coral; The method for preparing the gold nanorod includes the following steps: A gold salt solution is mixed with citrate and hydroxylamine, and then mixed with gold nano corals under stirring conditions. The reaction yields gold nano rosettes. The reaction time is 20–150 min; the final concentration of hydroxylamine in the reaction system is 0.01–0.1 M.

2. The gold nanogarland according to claim 1, characterized in that, The preparation process of the gold seed solution includes: mixing a reducing agent with a gold salt solution to obtain the gold seed solution.

3. The gold nanogarland according to claim 2, characterized in that, The reducing agent is selected from at least one of citrate, ascorbic acid, sodium borohydride and hydroxylamine; and / or, the gold salt is at least one of chloroauric acid and chloroaurate.

4. The gold nanogarland according to claim 3, characterized in that, The reducing agent in the preparation of the gold nanocoral includes citrate, sodium borohydride and hydroxylamine; and / or, the reducing agent in the preparation of the gold seed solution includes citrate and hydroxylamine.

5. The use of the gold nanorod according to any one of claims 1 to 4 in at least one of (1) to (5): (1) To prepare products for the treatment and / or prevention of liver diseases; (2) Drug delivery; (3) Preparation of a drug delivery system; (4) Preparation of drug carriers; (5) Prepare products for monitoring the distribution and / or location of drugs.

6. The application according to claim 5, characterized in that, The liver diseases described in (1) include acute liver injury.

7. A drug comprising the gold nanogarland according to any one of claims 1 to 4.

8. A drug delivery system, the gold nanorod of any one of claims 1 to 4, and a drug loaded on the gold nanorod.

Images