Preparation method and application of gold nanoparticle garland with acute liver injury treatment capacity

Abstract

The invention belongs to the technical field of biological medicine, and discloses a preparation method and application of gold nano garland with the acute liver injury treatment capacity. In particular to a gold nano garland, and hydroxylamine is loaded in the gold nano garland. The hydroxylamine is stably loaded in the gold nano garland, so that the biocompatibility of the hydroxylamine is improved, and the treatment effect of the hydroxylamine is enhanced through the characteristics of the gold nano garland, so that targeting and slow release are integrated, and potential biological efficiency is shown. The gold nano garland can specifically target liver cells, and systemic side effects related to conventional treatment can be reduced. The targeting method can enhance the effect of the therapeutic agent carried by the nanoparticles.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to a preparation method and application of a gold nano "roset" capable of treating acute liver injury. Background Art

[0002] Gold, as a metal element with very stable chemical properties, 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 intensity can be regulated, making them widely used in 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 reaction of various materials. However, the biocompatibility of hydroxylamine is extremely poor, which limits its application in living bodies.

[0003] Therefore, this application is specially filed. Summary of the invention

[0004] The present invention aims to solve at least one of the technical problems existing in the prior art. To this end, the present invention proposes a gold nano "rosette" (Au nanowreaths, AuNwrs) with the ability to treat acute liver injury. Through research, a method for stably loading hydroxylamine in the gold nanorosette is proposed, which not only improves the biocompatibility of hydroxylamine, but also enhances its therapeutic effect through the characteristics of the gold nanorosette, thereby integrating targeting and sustained release, showing potential biological efficacy. By stably loading hydroxylamine in the gold nanorosette, a new diagnosis and treatment idea can be provided for the treatment of liver injury in clinical practice and the blocking of the process of acute liver injury to chronic liver fibrosis.

[0005] The first aspect of the present invention aims to provide a gold nanorosette.

[0006] The purpose of the second aspect of the present invention is to provide a method for preparing the gold nanorosettes of the first aspect of the present invention.

[0007] The purpose of the third aspect of the present invention is to provide an application of the gold nanorosette of the first aspect of the present invention.

[0008] The fourth aspect of the present invention aims to provide a medicine.

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

[0010] In order to achieve the above object, the technical solution adopted by the present invention is:

[0011] The first aspect of the present invention provides a gold nanorosette, wherein the nanorosette is loaded with hydroxylamine.

[0012] In some embodiments of the present invention, the gold nanorosette is obtained by self-assembly of gold nanocorals through intermolecular hydrogen bonds of hydroxylamine.

[0013] In some embodiments of the present invention, the gold nanocoral is prepared by the following preparation method: mixing a gold salt solution with a reducing agent and a gold seed solution, and reacting them 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 a 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 process 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, and stirring until the solution turns pink, thereby obtaining the gold seed solution.

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

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

[0022] By adjusting the reaction conditions during the preparation of gold nanocorals, gold nanocorals with different diameters can be prepared. As the stirring time increases, the diameter of the gold nanocorals continues to expand.

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

[0024] The present invention provides a gold nanoroset specifically designed for targeted targeting of hepatocytes, aiming to provide a new idea for clinical treatment of acute liver damage and prevention of liver fibrosis. This new nanoroset itself has considerable biocompatibility and antioxidant capacity. In addition, by changing the surface of gold nanoparticles, new nanorosets with different particle sizes and carrying different drugs can be customized to regulate inflammatory responses in different organs, which is crucial in acute organ damage. These nanoparticles not only have inherent anti-inflammatory properties, but can also directly deliver antioxidants to hepatocytes, thereby more effectively reducing oxidative stress than traditional systemic antioxidant therapy and avoiding excessive side effects.

[0025] The gold nanoroset provided by the present invention has good biocompatibility and a longer metabolic cycle in the body than most clinical drugs. It can correct the inflammation and oxidative stress products that appear in acute liver injury in a long-term manner, reduce tissue damage, and treat acute liver injury and block the process of inflammatory transformation of acute liver injury to fibrosis.

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

[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 with gold nanocorals under stirring conditions, reacting to obtain gold nanoroselets.

[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 hydroxylamine in the reaction system is 0.01-0.1M.

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

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

[0033] By adjusting the concentration of hydroxylamine in the reaction system, the diameter of the gold nanorosette can be controlled.

[0034] Hydrogen chloroaurate (HAuCl4) provides gold ions for NaBH4 reduction. During the reduction process, these ions form gold atoms that aggregate into nanoparticles, which serve as the main material for the nanocoral structure. Hydroxylamine (NH2OH-HCl) can be used to reduce and generate gold nanorosettes on the one hand, and to control the growth kinetics of the gold nanorosette structure on the other hand. It provides a different reduction potential from NaBH4 and can be used to manipulate the shape and surface properties of gold nanoparticles. This method uses the unique properties of each reagent to control the nucleation and growth stages of gold nanoparticles to form complex nanorosette structures with potential high-impact applications.

[0035] The preparation method of the gold nanorosette provided by the invention is simple to operate, and the whole process does not need to use any reagents other than sodium borohydride, hydrogen tetrachloroaurate, trisodium citrate, and hydroxylamine. The method has high repeatability and low cost, and the gold nanorosette has a hollow structure, uniform size, good dispersibility, and the size and wall thickness can be adjusted.

[0036] The third aspect of the present invention provides the use of the gold nanorosette of the first aspect of the present invention in at least one of (1) to (5):

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

[0038] (2) Drug delivery;

[0039] (3) preparing drug delivery systems;

[0040] (4) preparing drug carriers;

[0041] (5) Prepare products for monitoring the distribution and / or localization 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 acute liver injury induced by carbon tetrachloride.

[0044] The gold nanorosettes provided by the present invention, characterized by their biocompatibility and non-toxicity at specific concentrations, provide patients with impaired liver function with a safer alternative to certain drugs that may have serious side effects. In addition, the unique properties of gold allow these nanoparticles to be imaged and monitored in real time in the body using technologies such as CT scanning or MRI, thereby accurately tracking the distribution of treatment and adjusting treatment strategies based on real-time data. The versatility of gold nanorosettes can also be used in combination with other treatment methods (such as drug therapy or gene therapy) to create synergistic effects and enhance the overall therapeutic effect. This multifunctional and adaptable nanoparticle system represents a major advancement in targeted treatment of acute organ damage, improving the effectiveness and safety of treatment methods.

[0045] The fourth aspect of the present invention provides a drug, which includes the gold nanorose ring of the first aspect of the present invention.

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

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

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

[0049] A fifth aspect of the present invention provides a drug delivery system, which includes the gold nanoroset according to the first aspect of the present invention, and a drug loaded on the gold nanoroset.

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

[0051] The beneficial effects of the present invention are:

[0052] The present invention stably loads hydroxylamine in gold nanorosettes, which not only improves the biocompatibility of hydroxylamine, but also enhances its therapeutic effect through the characteristics of gold nanorosettes, thereby integrating targeting and sustained release, and showing potential biological efficacy. The gold nanorosettes can specifically target liver cells, which may reduce the systemic side effects associated with conventional treatments. This targeting method can enhance the effect of the therapeutic agent carried by the nanoparticles.

[0053] Furthermore, the nano rosette provided by the present invention has the following characteristics:

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

[0055] Antioxidant properties: Oxidative stress plays a key role in the progression of liver damage. Gold nanorosettes can directly deliver and release antioxidants to damaged liver cells, 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 nanorosetties loaded with hydroxylamine can greatly improve the biocompatibility of hydroxylamine.

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

[0058] Combination therapy: Gold nanorosetties can be used in combination with other treatments, such as drug therapy or gene therapy, to create a synergistic effect and enhance the overall therapeutic effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0059] Figure 1 Flow chart for the preparation of AuNCls and AuNwrs.

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

[0061] Figure 3 TEM electron microscope images of AuNCls after different reaction times when the reactant ratio is Ratio 1; among them, a is the TEM electron microscope image with a reaction time of 30 min; b is the TEM electron microscope image with a reaction time of 60 min; c is the TEM electron microscope image with a reaction time of 90 min; d is the TEM electron microscope image with a reaction time of 120 min; e is the TEM electron microscope image with a reaction time of 150 min; f~j are high-resolution TEM electron microscope images corresponding to a~e; k is the XRD spectrum of AuNCls of different sizes.

[0062] Figure 4Schematic diagram of the synthesis of AuNwrs and TEM electron micrographs of AuNwrs of different sizes; among them, a is a schematic diagram of the synthesis of AuNwrs; b is a TEM electron micrograph of 58nm AuNwrs; c is a TEM electron micrograph of 79nm AuNwrs; d and e are TEM electron micrographs of 92nm AuNwrs; f is a TEM electron micrograph of 148nm AuNwrs; g is a TEM electron micrograph of 274nm AuNwrs; h is a TEM electron micrograph of 360nm AuNwrs.

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

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

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

[0066] Figure 8 Structural comparison of pathological sections of liver tissue under HE staining at 200x magnification in mice with carbon tetrachloride-induced liver damage model before and after treatment with silymarin and gold nanorose wreath. DETAILED DESCRIPTION

[0067] The present invention is further described in detail below through specific examples.

[0068] It should be understood that these embodiments are only used to illustrate the present invention and are not used to limit the scope of the present invention.

[0069] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0070] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0071] Example 1

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

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

[0074] Prepare 20 mL of 2.5 × 10 -4 M hydrogen tetrachloroaurate (HAuCl4) and 2.5×10 -4 0.6 mL of 0.1 M sodium borohydride (NaBH4) solution freshly prepared after freezing was added to 0.4 M trisodium citrate (Na3cit) aqueous solution, and the mixture was stirred continuously until the solution turned pink, indicating the formation of gold seeds, thus obtaining gold seeds (referred to as seed solution);

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

[0076] First, prepare 2.5 × 10 -4 200 mL of HAuCl4 solution was prepared, and 6 g of Na3cit was added thereto (final concentration: 0.08 M). The mixture was reacted at 35°C until the solution turned orange. The collected solution was cooled to room temperature and recorded as the growth stock solution.

[0077] Subsequently, a 50 mL conical flask was prepared, and 7.5 mL of the growth stock solution was mixed with 0.05 mL of freshly prepared 0.1 M NH2OH·HCl solution. 3.5 mL of the seed solution obtained in step (1) was added, and the mixture was stirred at 85°C for 30 min. The solution turned into wine red, and AuNCls were obtained.

[0078] TEM image analysis showed that the diameter of the AuNCls was 23 nm.

[0079] Example 2

[0080] A method for preparing gold nanocorals, which is different from Example 1 only in that the reaction conditions in the preparation process 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 measured to be 48 nm.

[0082] Example 3

[0083] A method for preparing gold nanocorals, which is different from Example 1 only in that the reaction conditions in the preparation process 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 nanocorals, which is different from Example 1 only in that the reaction conditions in the preparation process 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 measured to be 35 nm.

[0088] Example 5

[0089] A method for preparing gold nanocorals, which is different from Example 1 only in that the reaction conditions in the preparation process 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 measured to be 43 nm.

[0091] Example 6

[0092] A gold nanorod (AuNwrs) with antioxidant properties is formed by self-assembly of the gold nanorods prepared in Example 1 through intermolecular hydrogen bonds between NH2OH·HCl.

[0093] The preparation method of the above-mentioned gold nanorods (AuNwrs) comprises the following steps: taking 9 mL of the growth stock solution (preparing 2.5×10 -4 200 mL of 0.1 M HAuCl4 solution was prepared, and 6 g of Na3Cit (final concentration 0.08 M) was added thereto. The solution was reacted at 35°C until the color of the solution turned orange, and the collected solution was cooled to room temperature) and mixed with 0.45 mL of 0.1 M NH2OH·HCl solution, and 1.0 mL of AuNCls of Example 1 was added under vigorous stirring. After reacting at room temperature for 30 min, AuNwrs was obtained.

[0094] Through high-resolution TEM electron microscope image analysis, the diameter of the AuNwrs was found to be 148 nm.

[0095] Example 7

[0096] A gold nanocoral (AuNwrs) with antioxidant properties, which is different from Example 6 only in that the gold nanocoral is prepared in Example 2 and the amount of 0.1M NH2OH·HCl solution used in the preparation of AuNwrs is different (replaced from 0.45mL of 0.1M NH2OH·HCl solution to 10.8mL of 0.1M NH2OH·HCl solution).

[0097] High-resolution TEM image analysis showed that the diameter of AuNwrs was 360 nm.

[0098] Example 8

[0099] A gold nano "roset" (AuNwrs) with antioxidant properties, which is different from Example 6 only in that the gold nanocoral is prepared in Example 3.

[0100] High-resolution TEM image analysis showed that the diameter of the AuNwrs was 125 nm.

[0101] Example 9

[0102] A gold nano "roset" (AuNwrs) with antioxidant properties, which is different from Example 7 only in that the gold nanocoral is prepared in Example 4.

[0103] High-resolution TEM image analysis showed that the diameter of the AuNwrs was 150 nm.

[0104] Example 10

[0105] A gold nanocoral (AuNwrs) with antioxidant properties, which is different from Example 6 only in that the gold nanocoral is prepared in Example 5, and the reaction time at room temperature during the preparation of AuNwrs is 150 minutes.

[0106] High-resolution TEM image analysis showed that the diameter of the AuNwrs was 250 nm.

[0107] Embodiment 11

[0108] An antioxidant gold nanocoral (AuNwrs) is different from Example 10 in that Cy7 dye is added during the preparation process. The gold nanocoral is prepared in Example 1. Specifically, 9 mL of the growth stock solution is 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 in Example 1 are added under vigorous stirring. After reacting for 150 min, AuNwrs (denoted as Cy7-AuNwrs) is obtained.

[0109] Through high-resolution TEM electron microscope image analysis, the diameter of the AuNwrs was found to be 148 nm.

[0110] Example 12

[0111] A gold nanorod (AuNwrs) with antioxidant properties, which is different from Example 11 only in that the amount of 0.1M NH2OH·HCl solution used in the preparation process of AuNwrs is 10.8 mL.

[0112] High-resolution TEM image analysis showed that the diameter of the AuNwrs was 360 nm.

[0113] Effect Example

[0114] 1. Preparation and Characterization of Gold Nanocorals and Gold Nanorosettes

[0115] Figure 1 The preparation process of AuNCls and AuNwrs is shown in the figure. As shown in the figure, AuNCls are prepared from the bottom to the top in a two-step process. First, gold seeds (i.e., gold nanodots, Aunanodots, AuNDs) are synthesized using NaBH4 as a reducing agent. Figure 2 As shown in a and b, the gold seeds are nanodot structures of about 7.5 nm. Figure 2 Figure c is the selected area electron diffraction (SAED) pattern of AuNDs. It can be seen from the figure that the diffraction peak of the Au (111) plane is the strongest. The energy of this crystal plane is relatively low and it is the main place for subsequent secondary growth.

[0116] Subsequently, AuNDs were mixed with HAuCl4, Na3cit and the reducing agent NH2OH·HCl. As the reaction time increased, the size and number of AuNDs coral branches increased, showing a multi-branched structure. Figure 3 As shown in 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 The high-resolution TEM images in f to j show that the AuNCls obtained at different growth times all have lattice fringes with lattice spacing of 0.235nm and 0.203nm, which is consistent with the lattice structure of Au(111) and Au(100) crystal planes. This result shows that the series of nanoparticles obtained by changing the reaction time have the same crystal orientation and lattice structure. Further XRD diffraction of AuNCls is performed, and the results are as follows Figure 3As shown in figure k, AuNCls exhibit XRD diffraction peaks at 38.19°, 44.39°, 64.58°, 77.57° and 81.72°, which are attributed to Au (111), (200), (220), (111) and (420) planes, respectively. This result is consistent with the results shown by the above lattice fringes.

[0117] like Figure 4 As shown in b to d, as N H2 By changing the amount of NH2OH·HCl, the ratio of the reactants can be changed. Figure 4 As can be seen from Figures e to f, at this time, AuNCls (Example 2) are used as primitives to assemble into AuNWrs structures with a size of about 148 to 360 nm in the reaction system. The above results show that the size of AuNWrs can be regulated by changing the reaction time.

[0118] The above conclusions indicate that the driving force for the self-assembly of AuNWrs is the intermolecular hydrogen bonds formed between the excess NH2OH·HCl. When the amount of NH2OH·HCl is insufficient and the amount of Na3cit is large, since the -COOH in Na3cit exists in the form of -COO- in the solution, the electrostatic repulsion between them can maintain the existence of AuNCls in the form of single particles in the solution. As the amount of NH2OH·HCl gradually increases, intermolecular hydrogen bonds will form between them, thereby initiating the assembly of AuNCls, and the excess hydroxylamine will be adsorbed in the gold nanorosettes and released as the gold nanorosettes degrade.

[0119] Density functional theory (DFT) was further used to explore the properties of NH2OH·HCl and Na3cit on the gold surface. The calculation results show that the relative adsorption energy of Na3cit on Au(100) is 3.59 kJ / mol, which is higher than its relative adsorption energy on Au(111) crystal surface (3.52 kJ / mol). The adsorption of NH2OH·HCl on Au(100) and Au(111) is just the opposite ( Figure 3 These results indicate that NH2OH·HCl is more likely to attach to the Au(111) surface, while Na3cit is more likely to adsorb to the Au(100) surface, which allows the two to coexist on the surface of AuNCls. The intermolecular hydrogen bonds of Na3cit drive the self-assembly, and the electrostatic repulsion between Na3cit helps to maintain the hollow structure inside AuNwrs.

[0120] 2. Study on kidney targeting of AuNwrs

[0121] The premise for nanocarriers to achieve efficient treatment is to have good organ targeting. Here, male C57BL / 6 mice were selected as experimental mice, and 0.5 mg / kg AuNwrs labeled with Cy7 (i.e., AuNwrs of Example 11 and Example 12, 100 nm AuNwrs and 400 nm AuNwrs, respectively) were administered by tail vein injection. The mice were killed (euthanized) at different time points and their hearts, livers, spleens, lungs, kidneys, thymus, small intestines, muscles and brains were taken for in vitro organ fluorescence imaging.

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

[0123] The heart, liver, spleen, lung and other tissues of mice injected with 100 nm AuNwrs by tail vein were collected 7 days later for HE staining. Figure 6 shown.

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

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

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

[0127] The results are as follows Figure 7 As shown in Figure 2, the mortality rate of mice in the model group during the 7-day liver injury experiment was much higher than that in the treatment group (i.e., model + Yangshen control group and model + experimental group) and the sham group ( Figure 7 A), where gold nanoroset had an effective therapeutic effect on carbon tetrachloride-induced liver injury, and its NO, IL-6, ALT, AST and bilirubin indicators were similar to those of the positive control group ( Figure 7 From Figure 8 The HE staining results also show that both the positive control group and the nano-roset 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 modeling group mice.

[0128] In summary, the present invention provides a gold nanoroset specially designed to use a specific particle size and morphology for targeted targeting of hepatocytes, aiming to provide a new idea for the clinical treatment of acute liver damage and the prevention of liver fibrosis. This new nanoroset itself has considerable biocompatibility and antioxidant capacity. In addition, by utilizing the passive targeting principle that different organs take up different nanoparticle sizes, by changing the surface of gold nanoparticles, new nanorosets with different particle sizes and carrying different drugs can be customized to regulate the inflammatory response in different organs, which is crucial in acute organ damage. These nanoparticles not only have inherent anti-inflammatory properties, but can also directly deliver antioxidants to hepatocytes, thereby more effectively reducing oxidative stress than traditional systemic antioxidant therapy and avoiding excessive side effects.

[0129] The gold-based nanosystem used in the present invention, called gold nanorosettes, is characterized by its biocompatibility and non-toxicity at specific concentrations, providing patients with impaired liver function with a safer alternative to certain drugs that may have serious side effects. In addition, the unique properties of gold allow these nanoparticles to be imaged and monitored in real time in the body using technologies such as CT scans or MRI, allowing precise tracking of therapeutic distribution and adjustment of treatment strategies based on real-time data.

[0130] In addition, the present invention also utilizes the multifunctionality of gold nanorosettes and combines them with other treatments (such as drug therapy or gene therapy) to create a synergistic effect and enhance the overall therapeutic effect. This multifunctional and adaptable nanoparticle system represents a major advancement in targeted organ acute injury therapy, improving the efficacy and safety of treatments.

[0131] The embodiments of the present invention are described in detail above in conjunction with the accompanying drawings, but the present invention is not limited to the above embodiments. Various changes can be made within the knowledge of ordinary technicians in the relevant technical field without departing from the purpose of the present invention. In addition, the embodiments of the present invention and the features in the embodiments can be combined with each other without conflict.

Claims

1. A gold nanoroset, characterized in that: The gold nanorosette is loaded with hydroxylamine.

2. The gold nanorosette according to claim 1, characterized in that: The gold nanorosette is obtained by self-assembly of gold nanocorals through intermolecular hydrogen bonds of hydroxylamine.

3. The gold nanorosette according to claim 2, characterized in that: The gold nanocoral is prepared by the following preparation method: mixing a gold salt solution with a reducing agent and a gold seed solution, and reacting them to obtain a gold nanocoral; Preferably, the preparation process of the gold seed solution comprises: mixing a reducing agent with a gold salt solution to obtain a gold seed solution.

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

5. The gold nanorosette according to claim 3, characterized in that: The reaction conditions are stirring at 60-90° C. for 20-200 min.

6. The method for preparing the gold nanorosette according to any one of claims 2 to 5, comprising the following steps: The gold salt solution is mixed with citrate and hydroxylamine, and then mixed with the gold nanocorals under stirring conditions to react to obtain the gold nanocorals; Preferably, the reaction time is 20 to 150 min; and / or the final concentration of hydroxylamine in the reaction system is 0.01 to 0.1 M.

7. Use of the gold nanorosette according to any one of claims 1 to 5 in at least one of (1) to (5): (1) Preparation of products for the treatment and / or prevention of liver diseases; (2) Drug delivery; (3) preparing drug delivery systems; (4) preparing drug carriers; (5) Prepare products for monitoring the distribution and / or localization of drugs.

8. The use according to claim 7, characterized in that: The liver diseases mentioned in (1) include acute liver injury.

9. A medicine comprising the gold nanorosette according to any one of claims 1 to 5.

10. A drug delivery system, comprising the gold nanoroset according to any one of claims 1 to 5, and a drug loaded on the gold nanoroset.

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