NIR light-controlled precise drug release and ROS dynamic regulation phase change nano-particles and preparation method and application thereof

Abstract

The invention belongs to the field of biomedical materials and nano medicine, and discloses NIR light-controlled precise drug release and ROS dynamic regulation phase change nanoparticles as well as a preparation method and application of the NIR light-controlled precise drug release and ROS dynamic regulation phase change nanoparticles. According to the technology, a double-fatty-acid phase-change material is adopted as near-infrared response gating, composite gold nanoflower particles of a negative drug are wrapped through a one-pot method, targeting is given to the composite gold nanoflower particles through polyethyleneimine, and the gold-based phase-change nanoflower is constructed. The gold-based phase change nanoflowers can accurately regulate and control the drug release time sequence under NIR irradiation, and integrate efficient treatment and long-acting relapse control ability; meanwhile, active oxygen can be adjusted according to requirements, so that oxidative stress reaction is effectively adjusted, and local inflammation is remarkably relieved. And moreover, the preparation process is simple, the cost is low, and the advantages of excellent stability and biocompatibility are achieved, so that the preparation method has huge application potential and wide prospects in the biomedical fields of cancer treatment, anti-infection treatment, tissue repair and the like.

Description

Technical Field

[0001] The present invention relates to the field of biomedical materials and nanomedicine technology, and more specifically to a gold-based phase-change nanoflower capable of NIR light-controlled precise drug release and ROS dynamic regulation, and a preparation method and application thereof. Background Art

[0002] In the biomedical field, especially in key areas such as cancer treatment, anti-infection treatment and tissue repair, efficient, accurate and low-side effect treatment strategies have always been the goal pursued by researchers. Traditional treatment methods often find it difficult to find the best balance between treatment effects and side effects, especially in achieving precise drug release and regulating biological responses.

[0003] At present, the optimization and innovation of the performance of therapeutic materials for controlled drug release, as a key tool for achieving precision medicine in the fields of cancer treatment, anti-infection treatment and tissue repair, is the core driving force for the advancement of medical technology. However, such therapeutic materials often face many challenges when dealing with complex biological environments: first, the uncontrollable drug release leads to poor treatment effects and may aggravate the side effects of patients; second, the lack of an effective response mechanism to external stimuli (such as light, heat, etc.) limits the dynamic adjustment ability of the material during the treatment process; third, the stability and biocompatibility of the material in the body are still bottlenecks that restrict its widespread application.

[0004] Phase change materials (PCMs), as materials that can change physical states (such as solid-liquid phase transition) within a specific temperature range, have shown great potential in the field of drug control due to their unique temperature sensitivity and energy storage / release properties. However, single phase change materials usually lack active targeting functions and cannot effectively identify and locate diseased tissues or cells, resulting in widespread and unconcentrated distribution of drugs in the body, greatly reducing the therapeutic effect and possibly increasing damage to healthy tissues.

[0005] Although gold nanomaterials have strong drug-carrying capacity, their enzyme activity is relatively single. Carbon dots have attracted much attention in biomedicine due to their multi-enzyme activity, but their surface area limitations and modification problems have become bottlenecks. Researchers have cleverly combined the two to create a new type of composite gold nanoparticles: they retain the multi-enzyme activity of carbon dots, while enhancing drug loading and surface modification, to achieve drug diversity and therapeutic flexibility. However, despite their significant advantages, composite gold nanoparticles still have functional limitations and insufficient stability in drug delivery and biocatalysis. Its single function makes it difficult to cope with complex and changing treatment needs, and the double-edged sword effect of ROS - the right amount promotes treatment, while excessive amounts damage normal cells, induce inflammatory responses or tissue damage, which is a major challenge to treatment safety.

[0006] Therefore, how to effectively integrate the controlled release advantages of phase change materials with the multi-enzyme activity and drug loading capacity of composite gold nanoparticles to prepare a composite diagnostic and therapeutic material that integrates precise targeting, intelligent regulation, and efficient treatment and long-term recurrence control, so as to achieve efficient and accurate drug delivery and dynamic regulation of ROS, thereby ensuring the therapeutic effect while minimizing side effects, has become a key technical problem that technical personnel in this field need to overcome urgently. Summary of the invention

[0007] In view of this, the present invention provides a gold-based phase change nanoflower with NIR light-controlled precise drug release and ROS dynamic regulation, and a preparation method and application thereof. The present invention adopts a difatty acid phase change material (PCM) as a near-infrared (NIR) response gate, encapsulates the composite gold nanoflower particles of negative drugs through a one-pot method, and uses polyethyleneimine (PEI) to give it targeting, thereby constructing a technology of gold-based phase change nanoflowers.

[0008] One of the purposes of the present invention is to provide a method for preparing a gold-based phase-change nanoflower with NIR light-controlled precise drug release and ROS dynamic regulation, comprising the following steps:

[0009] (1) One-pot synthesis of gold nanoflowers with dopamine: Prepare a Tris-HCl buffer with a pH value of 8.5, add dopamine hydrochloride and chloroauric acid aqueous solution in sequence, stir at room temperature, centrifuge and resuspend to obtain AuNFs solution for later use;

[0010] (2) Alcohol-thermal method for synthesizing CDMn: o-phenylenediamine, dopamine, and potassium permanganate are dissolved in ethanol in sequence, and an alcohol-thermal reaction occurs at high temperature. The reaction is then dialyzed and freeze-dried to obtain dry CDMn.

[0011] (3) Synthesis of AuNFs@CDMn complex: Take an appropriate amount of carbon dots and disperse them in dimethyl sulfoxide. Slowly add AuNFs solution while stirring. Stir the mixture at room temperature. Wash the mixture by centrifugation and resuspend it to obtain AuNFs@CDMn solution for later use.

[0012] (4) Hydrophilic modification: Take an appropriate amount of PEG and dissolve it in ethanol. Slowly add it to the AuNFs@CDMn solution while stirring. Stir the reaction and resuspend it by centrifugation to obtain a hydrophilic AuNFs@CDMn solution for later use.

[0013] (5) One-pot loading of di-fatty acid: Take an appropriate amount of target drug X and dissolve it in deionized water, and slowly add the hydrophilic AuNFs@CDMn solution, stir and react to obtain AuNFs@CDMn@X solution; Take a certain amount of lauric acid and stearic acid and stir and dissolve them at high temperature, add the AuNFs@CDMn@X solution, stir and react quickly, then cool it down quickly in a cold water bath, stir and react at room temperature, centrifuge and resuspend, and obtain P@AMX solution for use;

[0014] (6) Enhance hydrophilicity: Take an appropriate amount of lecithin and PEG and dissolve them in ethanol, stir and react, slowly add P@AMX solution, stir and react, centrifuge and resuspend to obtain a hydrophilic P@AMX solution for later use;

[0015] (7) Imparting positive charge: Prepare PEI aqueous solution, slowly add hydrophilic P@AMX solution, stir to react, obtain P@AMX-P gold-based phase change nanoflowers, centrifuge and wash several times, and store at 4°C.

[0016] Furthermore, in step (1), the solid-liquid ratio of the dopamine hydrochloride to the Tris-HCL buffer is 1 g: 500-1000 mL, the mass percentage of chloroauric acid in the chloroauric acid aqueous solution is 1%-10%, and the mass percentage of the dopamine hydrochloride to the chloroauric acid is 2:1-1:2; the stirring at room temperature is 9-12 h, the stirring speed is 400-600 r / min, the centrifugal speed is 3500-6500 r / min, the centrifugal time is 10-20 min, and the concentration of the AuNFs solution is 0.5-1.8 mg / mL.

[0017] Furthermore, in step (2), the mass ratio of o-phenylenediamine, dopamine and potassium permanganate is 1:0.4-1:0.2-0.8, the solid-liquid ratio of o-phenylenediamine to ethanol solution is 1:15-45mL; the alcohol thermal reaction temperature is 140-220°C, the reaction time is 10-14h, the dialysis time is 12-24h, and the freeze-drying time is 72h.

[0018] Furthermore, in step (3), the solid-liquid ratio of the carbon dots to dimethyl sulfoxide is 1 mg: 5-10 mL; the mass ratio of the carbon dots to the AuNFs is 1: 1-1: 10; the room temperature stirring reaction time is 2-12 h, the centrifugal speed is 3500-6500 r / min, the centrifugal time is 10-20 min, and the concentration of the AuNFs@CDMn solution is 0.5-1.8 mg / mL.

[0019] Furthermore, in step (4), the solid-liquid ratio of the PEG to the ethanol is 1 mg:10-15 mL; the mass ratio of the PEG to the AuNFs@CDMn is 1:1-1:10; the stirring reaction time is 6-24 h, the stirring speed is 400-600 r / min, the centrifugal speed is 3500-6500 r / min, the centrifugal time is 10-20 min, and the concentration of the hydrophilic AuNFs@CDMn solution is 0.5-1.8 mg / mL.

[0020] Furthermore, in step (5), the concentration of the target drug X in deionized water is 0.5-1.5 mg / mL, the mass ratio of the hydrophilic AuNFs@CDMn to the target drug X is 1:2-1:6, the mass ratio of the lauric acid to the stearic acid is 1:1-8:1, and the mass ratio of the AuNFs@CDMn to the lauric acid is 1:10-1:25; the high temperature is 45-60°C, the stirring and dissolving time is 10-30 min, the stirring reaction time at room temperature is 1-3 h, the centrifugal speed is 3500-6500 r / min, the centrifugal time is 10-20 min, and the concentration of the P@AMX solution is 0.5-1.8 mg / mL.

[0021] Furthermore, in step (6), the mass ratio of the lecithin to the PEG is 1 to 10:1, the solid-liquid ratio of the PEG to the ethanol is 1 mg:1 to 5 mL, and the mass ratio of the lecithin to the P@AMX is 1:0.5 to 5; the stirring reaction time is 6 to 24 h, the centrifugal speed is 3500 to 6500 r / min, the centrifugal time is 10 to 20 min, and the concentration of the hydrophilic P@AMX solution is 0.5 to 1.8 mg / mL.

[0022] Furthermore, in step (7), the mass percentage of the PEI aqueous solution is 25% to 50%, the mass fraction ratio of the PEI to the hydrophilic P@AMV is 1:1 to 1:10; the stirring time is 12 to 48 hours, the centrifugal speed is 3500 to 6500 r / min, and the centrifugal time is 10 to 20 minutes.

[0023] The second purpose of the present invention is to provide a gold-based phase-change nanoflower with NIR light-controlled precise drug release and ROS dynamic regulation, wherein the size of the gold-based phase-change nanoflower is 100-180nm.

[0024] The third purpose of the present invention is to provide a gold-based phase change nanoflower with NIR light-controlled precise drug release and ROS dynamic regulation for use in biomedical materials and nanomedicine.

[0025] Furthermore, the gold-based phase-change nanoflowers can accurately control the timing of drug release under NIR irradiation, combining high-efficiency treatment with long-term inhibitory capabilities; at the same time, they can adjust the generation and clearance process of reactive oxygen species (ROS) according to demand, thereby effectively managing oxidative stress responses and significantly reducing local inflammation, so as to achieve applications in biomedical fields such as cancer treatment, infection control and tissue repair.

[0026] It can be seen from the above technical solution that, compared with the prior art, the beneficial effects of the present invention are:

[0027] This paper uses di-fatty acid phase change material (PCM) as near-infrared (NIR) response gate, encapsulates drug-negative composite gold nanoflower particles through a one-pot method, and uses polyethyleneimine (PEI) to give it targeting, thus constructing the technology of gold-based phase change nanoflowers. Under NIR irradiation, gold-based phase change nanoflowers can accurately control the timing of drug release, combining high-efficiency treatment and long-term inhibition capabilities; at the same time, they can adjust the generation and removal process of reactive oxygen species (ROS) according to demand, thereby effectively managing oxidative stress response and significantly reducing local inflammation. Its unique targeting, temperature sensitivity and photothermal synergistic therapy mechanism not only significantly enhance the therapeutic effect, but also greatly reduce the side effects of drugs. In addition, gold-based phase change nanoflowers have the advantages of simple preparation process, low cost, excellent stability and biocompatibility, which makes it show great application potential and broad prospects in biomedical fields such as cancer treatment, anti-infection treatment and tissue repair. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying creative work.

[0029] Figure 1 This is the TEM image of the P@AMV-P gold-based phase change nanoflower in Example 1.

[0030] Figure 2 This is the photothermal cycle heating diagram of the P@AMV-P gold-based phase change nanoflower in Example 1.

[0031] Figure 3 The drug release diagram of the P@AMV-P gold-based phase-change nanoflower in Example 1, including: (a) drug release under near-infrared on / off, and (b) drug cyclic release within four near-infrared on / off cycles.

[0032] Figure 4 This is the ROS generation diagram of the P@AMV-P gold-based phase change nanoflower in Example 1, including: (a) ROS generation under near-infrared on / off, and (b) ROS generation diagram under continuous near-infrared on / off state. DETAILED DESCRIPTION

[0033] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0034] Example 1: Preparation method of gold-based phase-change nanoflowers with NIR light-controlled precise drug release and ROS dynamic regulation

[0035] (1) Prepare 100 mL of Tris-HCl buffer with a pH value of 8.5, add 0.12 g of dopamine hydrochloride, and then add 0.8 mL of 1% chloroauric acid aqueous solution, stir at room temperature for 9 hours to obtain reduced nano-gold, centrifuge and resuspend for later use;

[0036] (2) 0.12 g of o-phenylenediamine, 0.08 g of dopamine, and 0.05 g of potassium permanganate were weighed and dissolved in 30 mL of ethanol in sequence, and the mixture was subjected to an alcohol thermal reaction at 160 °C for 6 h. The reaction solution was dialyzed for 12 h to remove unreacted small molecules and then freeze-dried to obtain dry carbon dot (CDMn) powder;

[0037] (3) Weigh 1 mg of carbon dot (CDMn) powder and dissolve it in 10 mL of dimethyl sulfoxide. Slowly add 10 mL of 1 mg / mL AuNFs solution while stirring and react for 6 h. After multiple centrifugation and washing, resuspend the solution for later use.

[0038] (4) Weigh 2 mg of polyethylene glycol (PEG) and dissolve it in 20 mL of ethanol solution. Slowly add 2 mL of 1 mg / mL AuNFs@CDMn solution while stirring. Stir the mixture for 12 h and resuspend by centrifugation for later use.

[0039] (5) Weigh 10 mg Van and dissolve it in 10 mL deionized water, then slowly add 2 mL of 1 mg / mL hydrophilic AuNFs@CDMn solution and stir for 6 h. Stir and dissolve 40 mg lauric acid and 10 mg stearic acid at 50 °C, then add AuNFs@CDMn@Van solution and stir rapidly at 50 °C for 20 min, then cool in a cold water bath for 3 min, and then stir and react at room temperature for 2 h. The resulting P@AMV solution is centrifuged and resuspended for later use.

[0040] (6) Weigh 15 mg of lecithin and 5 mg of PEG and dissolve them in 10 mL of ethanol and stir for 6 h. Then slowly add 10 mL of 1 mg / mL P@AMV solution and stir for 6 h. Centrifuge and resuspend for later use.

[0041] (7) Take 2 mL of 50% polyethyleneimine (PEI) aqueous solution, slowly add 2 mL of a hydrophilic P@AMV solution with a concentration of 1 mg / mL, stir and react for 24 hours, and after multiple centrifugation, wash and resuspension, the P@AMV-P gold-based phase change nanoflowers can be obtained and stored at 4°C.

[0042] The size and morphology of the gold-based phase-change nanoflowers with NIR light-controlled precise drug release and ROS dynamic regulation are as follows: Figure 1 As shown by Figure 1 It can be seen that the size of P@AMV-P gold-based phase change nanoflowers is 100-180nm.

[0043] Example 2: Preparation method of gold-based phase-change nanoflowers with NIR light-controlled precise drug release and ROS dynamic regulation

[0044] (1) Prepare 100 mL of Tris-HCl buffer with a pH value of 8.5, add 0.12 g of dopamine hydrochloride, then add 0.8 mL of 1% chloroauric acid aqueous solution, stir at room temperature for 9 h to reduce the nano-gold solution, and resuspend by centrifugation for later use;

[0045] (2) 0.12 g of o-phenylenediamine, 0.08 g of dopamine, and 0.05 g of potassium permanganate were weighed and dissolved in 30 mL of ethanol in sequence, and the mixture was subjected to an alcohol thermal reaction at 160 °C for 6 h. The reaction solution was dialyzed for 12 h to remove unreacted small molecules and freeze-dried to obtain dry carbon dot (CDMn) powder;

[0046] (3) Weigh 1 mg of carbon dot (CDMn) powder and dissolve it in 10 mL of dimethyl sulfoxide solution. Slowly add 10 mL of 1 mg / mL AuNFs solution while stirring and react for 6 h. After multiple centrifugation and washing, resuspend the solution for later use.

[0047] (4) Weigh 2 mg of polyethylene glycol (PEG) and dissolve it in 20 mL of ethanol solution. Slowly add 2 mL of 1 mg / mL AuNFs@CDMn solution while stirring. Stir the reaction for 12 h, and resuspend by centrifugation for later use.

[0048] (5) Weigh 10 mg of Kartogenin (KGN) and dissolve it in 10 mL of deionized water. Slowly add 2 mL of 1 mg / mL hydrophilic AuNFs@CDMn solution and stir for 6 h. Dissolve 40 mg of lauric acid and 10 mg of stearic acid at 50 °C. Then add AuNFs@CDMn@KGN solution and stir rapidly at 50 °C for 20 min. Then cool it in a cold water bath for 3 min. Then stir and react at room temperature for 2 h. The resulting P@AMK solution is centrifuged and resuspended for later use.

[0049] (6) Weigh 15 mg of lecithin and 5 mg of PEG and dissolve them in 10 mL of ethanol, stir and react for 6 h, then slowly add 10 mL of 1 mg / mL P@AMK solution, stir and react for 6 h, and centrifuge to resuspend for later use.

[0050] (7) Take 2 mL of 50% polyethyleneimine (PEI) aqueous solution, slowly add 2 mL of a hydrophilic P@AMK solution with a concentration of 1 mg / mL, stir and react for 24 hours, and after multiple centrifugation, wash and resuspension, the P@AMK-P gold-based phase change nanoflowers can be obtained and stored at 4°C.

[0051] Example 3: Preparation method of gold-based phase-change nanoflowers with NIR light-controlled precise drug release and ROS dynamic regulation

[0052] (1) Prepare 100 mL of Tris-HCl buffer with a pH value of 8.5, add 0.12 g of dopamine hydrochloride, then add 0.8 mL of 1% chloroauric acid aqueous solution, stir at room temperature for 9 h to reduce the nano-gold solution, and resuspend by centrifugation for later use;

[0053] (2) 0.12 g of o-phenylenediamine, 0.08 g of dopamine, and 0.05 g of potassium permanganate were weighed and dissolved in 30 mL of ethanol in sequence, and the mixture was subjected to an alcohol thermal reaction at 160 °C for 6 h. The reaction solution was dialyzed for 12 h to remove unreacted small molecules and freeze-dried to obtain dry carbon dot (CDMn) powder;

[0054] (3) Weigh 1 mg of carbon dot (CDMn) powder and dissolve it in 10 mL of dimethyl sulfoxide solution. Slowly add 10 mL of 1 mg / mL AuNFs solution while stirring and react for 6 h. After multiple centrifugation and washing, resuspend and set aside.

[0055] (4) Weigh 2 mg of polyethylene glycol (PEG) and dissolve it in 20 mL of ethanol solution. Slowly add 2 mL of 1 mg / mL AuNFs@CDMn solution while stirring. Stir the reaction for 12 h, and resuspend by centrifugation for later use.

[0056] (5) Weigh 10 mg of ampicillin (Amp) and dissolve it in 10 mL of deionized water, and slowly add 2 mL of a hydrophilic AuNFs@CDMn solution with a concentration of 1 mg / mL, and stir and react for 6 hours; 40 mg of lauric acid and 10 mg of stearic acid are stirred and dissolved at 50°C, and then the AuNFs@CDMn@Amp solution is added, and the reaction is rapidly stirred at 50°C for 20 minutes, then cooled in a cold water bath for 3 minutes, and then stirred and reacted at room temperature for 2 hours. The resulting P@AMA solution is centrifuged and resuspended for use.

[0057] (6) Weigh 15 mg of lecithin and 5 mg of PEG and dissolve them in 10 mL of ethanol, stir and react for 6 h, then slowly add 10 mL of 1 mg / mL P@AMA solution, stir and react for 6 h, and collect by centrifugation for later use.

[0058] (7) Take 2 mL of 50% polyethyleneimine (PEI) aqueous solution, slowly add 2 mL of a hydrophilic P@AMK solution with a concentration of 1 mg / mL, stir and react for 24 hours, and after multiple centrifugation, wash and resuspension, the P@AMA-P gold-based phase change nanoflowers can be obtained and stored at 4°C.

[0059] Example 4: Preparation method of gold-based phase-change nanoflowers with NIR light-controlled precise drug release and ROS dynamic regulation

[0060] (1) Prepare 100 mL of Tris-HCl buffer with a pH value of 8.5, add 0.12 g of dopamine hydrochloride, then add 0.8 mL of 1% chloroauric acid aqueous solution, stir at room temperature for 9 h to reduce the nano-gold solution, and resuspend by centrifugation for later use;

[0061] (2) 0.12 g of o-phenylenediamine, 0.08 g of dopamine, and 0.05 g of potassium permanganate were weighed and dissolved in 30 mL of ethanol in sequence, and the mixture was subjected to an alcohol thermal reaction at 160 °C for 6 h. The reaction solution was dialyzed for 12 h to remove unreacted small molecules and freeze-dried to obtain dry carbon dot (CDMn) powder;

[0062] (3) Weigh 1 mg of carbon dot (CDMn) powder and dissolve it in 10 mL of dimethyl sulfoxide solution. Slowly add 10 mL of 1 mg / mL AuNFs solution while stirring and react for 6 h. After multiple centrifugation and washing, resuspend the solution for later use.

[0063] (4) Weigh 2 mg of polyethylene glycol (PEG) and dissolve it in 20 mL of ethanol solution. Slowly add 2 mg / mL of 1 mg / mL AuNFs@CDMn solution while stirring. Stir the reaction for 12 h, and resuspend by centrifugation for later use.

[0064] (5) Weigh 10 mg of doxorubicin hydrochloride (DOX) and dissolve it in 10 mL of deionized water, then slowly add 2 mL of 1 mg / mL hydrophilic AuNFs@CDMn solution and stir for 6 h. Stir and dissolve 40 mg of lauric acid and 10 mg of stearic acid at 50 °C, then add AuNFs@CDMn@DOX solution and stir rapidly at 50 °C for 20 min, then cool in a cold water bath for 3 min, and then stir and react at room temperature for 2 h. The resulting P@AMD solution is centrifuged and resuspended for later use.

[0065] (6) Weigh 15 mg of lecithin and 5 mg of PEG and dissolve them in 10 mL of ethanol, stir and react for 6 h, then slowly add 10 mL of 1 mg / mL P@AMD solution, stir and react for 6 h, and centrifuge to resuspend for later use.

[0066] (7) Take 2 mL of 50% polyethyleneimine (PEI) aqueous solution, slowly add 2 mL of a hydrophilic P@AMD solution with a concentration of 1 mg / mL, stir and react for 24 hours, and after multiple centrifugation, wash and resuspension, the P@AMD-P gold-based phase change nanoflowers can be obtained and stored at 4°C.

[0067] The content of the present invention is not limited to the content of the above embodiments. The combination of one or more embodiments can also achieve the purpose of the present invention.

[0068] In order to further verify the excellent effect of the present invention, the inventors also conducted the following experiments:

[0069] Experiment 1: To illustrate the photothermal effect of P@AMV-P gold-based phase-change nanoflowers

[0070] In order to study the photothermal heating ability of P@AMV-P gold-based phase change nanoflowers under low power of 808nm near-infrared irradiation, 500μg / mL P@AMV-P was placed in a quartz dish with four sides transparent to light. The laser power was set to 1.5W / cm 2 . Use near-infrared light 808nm for 6 minutes, then turn off the near-infrared light, wait until the temperature drops to room temperature and turn on the near-infrared light again. Repeat this step three times. Use an infrared imager to record the temperature every 30 seconds. The temperature-time curve is as follows Figure 3 shown.

[0071] Depend on Figure 2 It can be seen that under laser illumination with a wavelength of 808nm, P@AMV-P exhibits photothermal and cyclic heating characteristics.

[0072] Experiment 2: Used to illustrate the drug release of P@AMV-P gold-based phase change nanoflowers under near-infrared on / off

[0073] In order to study the drug release of P@AMV-P gold-based phase change nanoflowers under near-infrared on / off, 4 mg of P@AMV-P was dissolved in 8 mL of PBS, and 4 mL of P@AMV-P solution was placed at room temperature. 1 mL of P@AMV-P solution was centrifuged at 0 min, 2 min, 4 min, and 6 min to collect the supernatant, and the Van content in the supernatant was detected using a UV spectrophotometer; 1 mL of P@AMV-P solution was taken and near-infrared (1.5 W / cm 2) for 0 min, 2 min, 4 min and 6 min respectively. After irradiation, the supernatant was collected by centrifugation immediately, and the Van content in the supernatant was detected by ultraviolet spectrophotometer.

[0074] Depend on Figure 3 (a) It can be seen that under the stimulation of near-infrared light, PCM undergoes a phase change and releases Van.

[0075] Experiment 3: To illustrate the drug release cycle of P@AMV-P gold-based phase change nanoflowers in four near-infrared on / off cycles

[0076] In order to study the cyclic release characteristics of drugs from P@AMV-P gold-based phase-change nanoflowers under continuous near-infrared light (NIR) on / off conditions, 10 mg of P@AMV-P gold-based phase-change nanoflowers were dissolved in 20 mL of PBS solution, and four groups of experiments were conducted, all of which started at the same time. Each group contained five samples, and the volume of each sample was 1 mL.

[0077] The first set of experiments: Five 1 mL samples were taken from the total solution. The first four samples were exposed to near infrared light (1.5 W / cm 2 ) for 0 min, 2 min, 4 min, and 6 min. Each sample was immediately centrifuged after irradiation to collect the supernatant. The fifth sample was also irradiated for 6 min and allowed to stand at room temperature for 6 min, then centrifuged to collect the supernatant, and the Van content in all the collected supernatants was measured using a UV spectrophotometer.

[0078] The second group of experiments: All five samples were first exposed to near-infrared light (1.5 W / cm 2 ) for 6 min. After the irradiation, all samples were left standing at room temperature for 24 h. The first four samples were irradiated again under near-infrared light for 0 min, 2 min, 4 min, and 6 min, and the supernatant was immediately collected by centrifugation. The fifth sample was also irradiated for 6 min and then left standing for 6 min and then centrifuged. The Van content in the supernatant was also measured using a UV spectrophotometer.

[0079] The third group of experiments: Repeat the initial steps of the second group of experiments, that is, all samples were irradiated with near-infrared light for 6 minutes and then left to stand for 24 hours. This process was repeated twice. Subsequently, the first four samples were irradiated with near-infrared light for 0 minutes, 2 minutes, 4 minutes, and 6 minutes, and the supernatant was immediately collected by centrifugation. The fifth sample was irradiated for 6 minutes, then left to stand for 6 minutes and then centrifuged. The Van content in the supernatant was detected using a UV spectrophotometer.

[0080] The fourth group of experiments: The initial and static steps of the third group were repeated, that is, all samples were subjected to three cycles of near-infrared irradiation for 6 minutes and then static for 24 hours. After that, the first four samples were subjected to near-infrared irradiation for 0 minutes, 2 minutes, 4 minutes, and 6 minutes and immediately centrifuged, and the fifth sample was irradiated for 6 minutes and then static for 6 minutes and then centrifuged. The Van content in each supernatant was measured by UV spectrophotometer to evaluate the cyclic release effect.

[0081] Depend on Figure 3 (b) It can be seen that P@AMV-P gold-based phase change nanoflowers can achieve drug cyclic release under multiple on / off stimulation of near-infrared light.

[0082] Experiment 4: used to illustrate the ROS generation of P@AMV-P gold-based phase change nanoflowers under near-infrared on / off

[0083] In order to study the ROS generation of P@AMV-P gold-based phase change nanoflowers under near-infrared on / off, 50ug / mL 100uL P@AMV-P solution was added to 100uL 100mM DMPO solution, mixed well, and sealed with a capillary tube. Then, the sample tube was placed in an electron spin resonance spectrometer for testing and data collection, divided into dark and near-infrared light (1.5W / cm 2 ) Group with 10 min illumination.

[0084] Depend on Figure 4 (a) It can be seen that P@AMV-P gold-based phase change nanoflowers can produce ROS under near-infrared light stimulation.

[0085] Experiment 5: To illustrate the ROS generation of P@AMV-P gold-based phase change nanoflowers under near-infrared continuous on / off

[0086] In order to study the ROS generation of P@AMV-P gold-based phase change nanoflowers under continuous near-infrared on / off, 50ug / mL 300uL P@AMV-P solution was added to a mixed solution of 360uL TMB solution (20mM), 100uL H2O2 solution (20mM), and 740uL acetic acid-sodium acetate solution (pH = 4.5), and the mixture was irradiated with near-infrared light (1.5W / cm 2 ) light, and use a UV spectrophotometer to detect the absorption value; then let it stand in the dark for 10 minutes, and use a UV spectrophotometer to detect the absorption value; repeat the above operation four times.

[0087] Depend on Figure 4 (b) It can be seen that P@AMV-P gold-based phase change nanoflowers can regulate the generation of ROS under continuous near-infrared light on / off, realizing dynamic regulation of ROS regulated by near-infrared light.

[0088] The various embodiments in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referenced to each other.

Claims

1. A method for preparing a gold-based phase-change nanoflower with NIR light-controlled precise drug release and ROS dynamic regulation, characterized in that: The following steps are involved: (1) One-pot synthesis of gold nanoflowers with dopamine: Prepare a Tris-HCl buffer with a pH value of 8.5, add dopamine hydrochloride and chloroauric acid aqueous solution in sequence, stir at room temperature, centrifuge and resuspend to obtain AuNFs solution for later use; (2) Alcohol-thermal method for synthesizing CDMn: o-phenylenediamine, dopamine, and potassium permanganate are dissolved in ethanol in sequence, and an alcohol-thermal reaction occurs at high temperature. The reaction is then dialyzed and freeze-dried to obtain dry CDMn. (3) Synthesis of AuNFs@CDMn complex: Take an appropriate amount of carbon dots and disperse them in dimethyl sulfoxide. Slowly add AuNFs solution while stirring. Stir the mixture at room temperature. Wash the mixture by centrifugation and resuspend it to obtain AuNFs@CDMn solution for later use. (4) Hydrophilic modification: Take an appropriate amount of PEG and dissolve it in ethanol. Slowly add it to the AuNFs@CDMn solution while stirring. Stir the reaction and resuspend it by centrifugation to obtain a hydrophilic AuNFs@CDMn solution for later use. (5) One-pot loading of di-fatty acid: Take an appropriate amount of target drug X and dissolve it in deionized water, and slowly add the hydrophilic AuNFs@CDMn solution, stir and react to obtain AuNFs@CDMn@X solution; Take a certain amount of lauric acid and stearic acid and stir and dissolve them at high temperature, add the AuNFs@CDMn@X solution, stir and react quickly, then cool it down quickly in a cold water bath, stir and react at room temperature, centrifuge and resuspend, and obtain P@AMX solution for use; (6) Enhance hydrophilicity: Take an appropriate amount of lecithin and PEG and dissolve them in ethanol, stir and react, slowly add P@AMX solution, stir and react, centrifuge and resuspend to obtain a hydrophilic P@AMX solution for later use; (7) Imparting positive charge: Prepare PEI aqueous solution, slowly add hydrophilic P@AMX solution, stir to react, obtain P@AMX-P gold-based phase change nanoflowers, centrifuge and wash several times, and store at 4°C.

2. The method for preparing a gold-based phase-change nanoflower with NIR light-controlled precise drug release and ROS dynamic regulation according to claim 1, characterized in that: In step (1), the solid-liquid ratio of the dopamine hydrochloride to the Tris-HCL buffer is 1g:500-1000mL, the mass percentage of chloroauric acid in the chloroauric acid aqueous solution is 1%-10%, and the mass percentage of the dopamine hydrochloride to the chloroauric acid is 2:1-1:2; the stirring is carried out at room temperature for 9-12h, the stirring speed is 400-600r / min, and the concentration of the AuNFs solution is 0.5-1.8mg / mL.

3. The method for preparing a gold-based phase-change nanoflower with NIR light-controlled precise drug release and ROS dynamic regulation according to claim 1, characterized in that: In step (2), the mass ratio of o-phenylenediamine, dopamine and potassium permanganate is 1:0.4-1:0.2-0.8, the solid-liquid ratio of o-phenylenediamine to ethanol solution is 1:15-45mL; the alcohol thermal reaction temperature is 140-220°C, the reaction time is 10-14h, the dialysis time is 12-24h, and the freeze-drying time is 72h.

4. The method for preparing a gold-based phase-change nanoflower with NIR light-controlled precise drug release and ROS dynamic regulation according to claim 1, characterized in that: In step (3), the solid-liquid ratio of the carbon dots to dimethyl sulfoxide is 1 mg:5-10 mL; the mass ratio of the carbon dots to the AuNFs is 1:1-1:10; the room temperature stirring reaction time is 2-12 h, and the concentration of the AuNFs@CDMn solution is 0.5-1.8 mg / mL.

5. The method for preparing a gold-based phase-change nanoflower with NIR light-controlled precise drug release and ROS dynamic regulation according to claim 1, characterized in that: In step (4), the solid-liquid ratio of the PEG to the ethanol is 1 mg:10-15 mL; the mass ratio of the PEG to the AuNFs@CDMn is 1:1-1:10; the stirring reaction time is 6-24 h, the stirring speed is 400-600 r / min, and the concentration of the hydrophilic AuNFs@CDMn solution is 0.5-1.8 mg / mL.

6. The method for preparing a gold-based phase-change nanoflower with NIR light-controlled precise drug release and ROS dynamic regulation according to claim 1, characterized in that: In step (5), the concentration of the target drug X in deionized water is 0.5-1.5 mg / mL, the mass ratio of the hydrophilic AuNFs@CDMn to the target drug X is 1:2-1:6, the mass ratio of the lauric acid to the stearic acid is 1:1-8:1, and the mass ratio of the AuNFs@CDMn to the lauric acid is 1:10-1:25; the high temperature is 45-60°C, the stirring and dissolving time is 10-30 min, the stirring reaction time at room temperature is 1-3 h, and the concentration of the P@AMX solution is 0.5-1.8 mg / mL.

7. The method for preparing a gold-based phase-change nanoflower with NIR light-controlled precise drug release and ROS dynamic regulation according to claim 1, characterized in that: In step (6), the mass ratio of the lecithin to the PEG is 1 to 10:1, the solid-liquid ratio of the PEG to the ethanol is 1 mg:1 to 5 mL, and the mass ratio of the lecithin to the P@AMX is 1:0.5 to 5; the stirring reaction time is 6 to 24 h, and the concentration of the hydrophilic P@AMX solution is 0.5 to 1.8 mg / mL.

8. The method for preparing a gold-based phase-change nanoflower with NIR light-controlled precise drug release and ROS dynamic regulation according to claim 1, characterized in that: In step (7), the mass concentration of the PEI aqueous solution is 25% to 50%, the mass ratio of the PEI to the hydrophilic P@AMX is 1:1 to 1:10; and the stirring time is 12 to 48 hours.

9. A gold-based phase-change nanoflower with NIR light-controlled precise drug release and ROS dynamic regulation as claimed in any one of claims 1 to 8, characterized in that: The size of the gold-based phase-change nanoflower is 100-180 nm.

10. Application of a gold-based phase-change nanoflower for NIR light-controlled precise drug release and ROS dynamic regulation, characterized in that: Application of the gold-based phase-change nanoflowers as claimed in claim 9 in biomedical materials and nanomedicine.

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