A near-infrared absorbing cerium-doped carbon quantum dot nanoscale enzyme, a preparation method and applications thereof

Abstract

The application provides a kind of near-infrared absorption cerium-doped carbon quantum dots nanoscale enzyme, preparation method and application, it is related to biomedical technology field, wherein the preparation method steps are as follows: S1: take hydrochloric acid dopamine, melamine, cerium nitrate, use anhydrous ethanol to soak thoroughly;S2: the mixture obtained in S1 is transferred to polytetrafluoroethylene lining hydrothermal reactor for reaction;S3: the solution in S2 is reacted with high-speed centrifugation, take supernatant and wash with anhydrous ethanol;After washing, dialysis is carried out with anhydrous ethanol, after dialysis, the solution in dialysis bag is spin-dried in a rotary evaporator, and the CeCDs solid is stored at-20 DEG C.The cerium-doped carbon quantum dots constructed by the application have good biocompatibility, can be applied to anti-parkinson disease, the carbon-based nanoscale enzyme CeCDs has SOD and CAT nanoscale enzyme catalytic activity, is used for the treatment of Parkinson's disease, and can pass through blood-brain barrier, and has good biocompatibility and treatment potential.

Description

Technical Field

[0001] This application relates to the field of biomedical technology, and in particular to a near-infrared absorbing cerium-doped carbon quantum dot nanozyme, its preparation method, and its applications. Background Technology

[0002] Parkinson's disease (PD) is a common neurodegenerative disorder characterized by the progressive loss of dopaminergic neurons in the substantia nigra of the midbrain and the abnormal accumulation of α-synuclein. Oxidative stress, mitochondrial dysfunction, and neuroinflammation are considered key factors in the pathogenesis of PD. Currently, treatment relies on drugs such as levodopa, but these drugs have difficulty crossing the blood-brain barrier (BBB), and most remain in the periphery, causing numerous side effects. Summary of the Invention

[0003] The purpose of this application is to solve the technical problem that existing drugs for treating Parkinson's disease have difficulty crossing the blood-brain barrier and causing side effects.

[0004] To address the aforementioned technical problems, this application provides the following technical solution:

[0005] A method for preparing a near-infrared absorbing cerium-doped carbon quantum dot biomimetic nanozyme includes the following steps:

[0006] S1: Weigh out dopamine hydrochloride, melamine, and cerium ammonium nitrate, and thoroughly soak them with anhydrous ethanol;

[0007] S2: Transfer the mixture obtained in S1 to a polytetrafluoroethylene-lined hydrothermal reactor for reaction;

[0008] S3: Centrifuge the solution from S2 after the reaction is complete at high speed, collect the supernatant and wash it with anhydrous ethanol;

[0009] After washing, the solution was dialyzed with anhydrous ethanol. After dialysis, the solution in the dialysis bag was dried by rotary evaporation to obtain CeCDs solid, which was stored at -20°C.

[0010] Preferably, the ratio of dopamine hydrochloride, melamine, cerium ammonium nitrate, and anhydrous ethanol in S1 is 2 g: 2 g: 1 g: 300 mL.

[0011] Preferably, the reaction conditions in the polytetrafluoroethylene-lined hydrothermal reactor in S2 are 180°C for 12 hours.

[0012] Preferably, the high-speed centrifugation conditions in S3 are 20,000 rpm for 15 min, followed by two washings after centrifugation, and the dialysis bag used for dialysis after washing has a retention mass of 1000 kDa.

[0013] This application also provides a near-infrared absorbing cerium-doped carbon quantum dot biomimetic nanozyme, which is prepared using the preparation method described above.

[0014] Preferably, the cerium-doped carbon quantum dot biomimetic nanozyme has a quantum dot structure with Ce-doped carbon dots as its core.

[0015] Preferably, the cerium-doped carbon quantum dot biomimetic nanozyme has nanozyme activity and can responsively produce hydrogen in response to 808nm near-infrared light.

[0016] This application also provides the application of the aforementioned near-infrared absorbing cerium-doped carbon quantum dot biomimetic nanozyme in the preparation of drugs for treating Parkinson's disease.

[0017] Preferably, the cerium-doped carbon quantum dot biomimetic nanozyme has SOD and CAT catalytic activity, which can reduce ROS generation.

[0018] Preferably, the nanozyme is an SOD,CAT nanozyme used for the treatment of Parkinson's disease.

[0019] Compared with the prior art, this application has at least the following beneficial effects:

[0020] This invention presents a method for preparing near-infrared absorbing cerium-doped carbon quantum dot nanozymes. The cerium-doped carbon quantum dots have a quantum dot structure, with carbon dots as the core and modified by cerium.

[0021] The cerium-doped carbon quantum dots constructed in this application have good biocompatibility, which is beneficial for their application in the field of biomedical technology. They also have nanozyme activity and can responsively produce hydrogen in response to 808nm near-infrared light.

[0022] The cerium-doped carbon quantum dots provided by this invention can be used to treat Parkinson's disease and represent a novel material. The preparation method of this invention is simple, has good reproducibility, and shows promising prospects for industrial production.

[0023] The carbon-based nanozymes CeCDs provided by this invention have SOD and CAT nanozyme catalytic activities and can be used for the treatment of Parkinson's disease.

[0024] The CeCDs provided by this invention can cross the blood-brain barrier and have good biocompatibility and therapeutic potential. Attached Figure Description

[0025] Figure 1 The image shows the X-ray diffraction (XRD) pattern of the synthesized product CeCDs according to one embodiment of the present invention.

[0026] Figure 2This is the ultraviolet absorption spectrum of cerium-doped carbon dots (CeCDs) in one embodiment of the present invention.

[0027] Figure 3 This is a transmission electron microscope (TEM) image of cerium-doped carbon dots (CeCDs) in one embodiment of the present invention.

[0028] Figure 4 This is a particle size distribution of cerium-doped carbon dots (CeCDs) in one embodiment of the present invention.

[0029] Figure 5 Evaluation of the SOD and CAT nanozyme catalytic performance of cerium-doped carbon dot nanozymes (CeCDs) in one embodiment of the present invention.

[0030] Figure 6 This is a near-infrared responsive hydrogen production diagram of cerium-doped carbon dots (CeCDs) in one embodiment of the present invention.

[0031] Figure 7 This is a CLSM imaging (400X) of cerium-doped carbon dots (CeCDs) in one embodiment of the present invention, showing the blood-penetrating brain barrier cell compartments.

[0032] Figure 8 This is a live imaging image of cerium-doped carbon dot nanozymes (CeCDs) according to one embodiment of the present invention. Detailed Implementation

[0033] This application provides a near-infrared absorbing cerium-doped carbon quantum dot biomimetic nanozyme, wherein the cerium-doped carbon quantum dot biomimetic nanozyme has a quantum dot structure with Ce-doped carbon dots as its core.

[0034] In one embodiment, the cerium-doped carbon quantum dot biomimetic nanozyme has nanozyme activity and can responsively produce hydrogen in response to 808 nm near-infrared light.

[0035] In one embodiment, the cerium-doped carbon quantum dot biomimetic nanozyme has SOD and CAT catalytic activities, which can reduce ROS generation.

[0036] In one embodiment, the cerium-doped carbon quantum dot biomimetic nanozyme can also be used to prepare drugs for treating Parkinson's disease.

[0037] The preparation method of the near-infrared absorbing cerium-doped carbon quantum dot biomimetic nanozyme is as follows:

[0038] S1: Weigh out dopamine hydrochloride, melamine, and cerium ammonium nitrate, dissolve them in anhydrous ethanol, and thoroughly soak them.

[0039] In one embodiment, the ratio of dopamine hydrochloride, melamine, cerium ammonium nitrate, and anhydrous ethanol is 2 g: 2 g: 1 g: 300 mL.

[0040] S2: Transfer the mixture obtained in S1 to a polytetrafluoroethylene-lined hydrothermal reactor for reaction.

[0041] The reaction conditions were 180°C for 12 h.

[0042] S3: Centrifuge the solution from S2 after the reaction is complete at high speed, take the supernatant and wash it twice with anhydrous ethanol; the high speed centrifugation conditions are 20000 rpm for 15 min.

[0043] After washing, the sample was dialyzed with anhydrous ethanol. The dialysis bag used for dialysis had a retention capacity of 1000 kDa.

[0044] After dialysis, the solution in the dialysis bag was evaporated in a rotary evaporator to obtain CeCDs solid, which was stored at -20°C.

[0045] The above content will be elaborated below with specific verification experiments:

[0046] I. Experimental Materials and Sources

[0047]

[0048] II. Verification Experiment

[0049] Example 1: Preparation of CeCDs

[0050] (1) Weigh 0.1 g of dopamine hydrochloride, 0.1 g of melamine, and 0.05 g of cerium ammonium nitrate, and wet them with 15 mL of anhydrous ethanol. Transfer them to a polytetrafluoroethylene-lined hydrothermal reactor and react at 180 °C for 12 h.

[0051] (2) After the reaction was completed, the solution was cooled to room temperature and centrifuged at 20,000 rpm for 15 min. The supernatant was collected and washed twice with anhydrous ethanol. The solution was dialyzed for 24 h using a dialysis bag with a cutoff mass of 1000 kDa. After the dialysis was completed, the solution was evaporated in a rotary evaporator to obtain CeCDs solid, which was stored at -20℃.

[0052] Please see Figure 1 and Figure 2 In this application, Ce was successfully doped into carbon dots without significant change in the crystal structure of the nanosheets; furthermore, the absorption of CeCDs in the NIR region was significantly enhanced. Figure 3 and Figure 4 It is evident that the obtained CeCDs have a particle size of less than 10 nm, which is beneficial for crossing the blood-brain barrier to treat Parkinson's disease.

[0053] Example 2: Verification of the catalytic activity of SOD and CAT nanozymes

[0054] In this application, the SOD and CAT catalytic activities of CeCDs nanozymes were evaluated using kits according to the instructions.

[0055] The results are as follows Figure 5 As shown, cerium-doped carbon dot nanozymes (CeCDs) exhibit SOD and CAT nanozyme catalytic activities.

[0056] Example 3: Verification of extracellular hydrogen production

[0057] SH-SY5Y cells were seeded in six-well plates and cultured for 24 h. The culture medium was then changed, and methylene blue was added and cultured for another 24 h. CeCDs were then added to a final concentration of 50 μg / mL and treated for 6 h. The changes in the intensity of methylene blue color within the cells were observed under a microscope.

[0058] Please see Figure 6 ,Depend on Figure 6 It is evident that the blue color in CeCDs-treated SH-SY5Y cells became lighter with increasing 808 nm laser irradiation time, indicating a positive correlation between H2 generation and irradiation time. The cells exhibited optimal hydrogen production after 15 minutes of laser irradiation.

[0059] Example 4: Simulated in vitro blood transfusion brain experiment

[0060] Bend.3 cells were seeded in the upper chamber of the cell culture chamber, and SH-SY5Y cells were seeded in the lower chamber. After a dense barrier was formed in the upper chamber by the Bend.3 cells, the cells were cultured in the upper chamber with 50 μg / mL RhB-labeled nanozymes (CeCDs) for 24 h. The cells in the lower chamber were fixed with 4% paraformaldehyde and gently washed three times with PBS to remove the fixative. Intracellular RhB fluorescence was observed using a confocal laser scanning microscope (CLSM).

[0061] Please see Figure 7 ,Depend on Figure 7 It was observed that after co-culturing for a period of time, RhB-labeled nanozymes CeCDs were able to cross Bend.3 cells and enter SH-SY5Y cells. This indicates that CeCD nanozymes can cross the blood-brain barrier.

[0062] Example 5: Mouse Experiment Validation

[0063] The RhB-labeled nanozymes CeCDs prepared in Example 1 of this invention, at a dose of 3 mg / kg, were injected via the tail vein into a mouse model of Parkinson's disease. Analysis was performed using an in vivo imaging system 6 h after tail vein injection.

[0064] Please see Figure 8 ,Depend on Figure 8It is evident that the cerium-doped carbon-based nanozymes (CeCDs) prepared in this invention can target brain regions and achieve specific treatment.

[0065] In summary, this application constructs a novel cerium-doped carbon quantum dot nanozyme with near-infrared light absorption, which has good biocompatibility and biometabolism, making it suitable for application in the biomedical field. Furthermore, the cerium-doped carbon quantum dot nanozyme can absorb near-infrared light. The cerium-doped carbon quantum dot nanozyme constructed in this application has a simple preparation method, good reproducibility, and good prospects for industrial production.

Claims

1. A method for preparing a near-infrared absorbing cerium-doped carbon quantum dot biomimetic nanozyme, characterized in that: Includes the following steps: S1: Weigh out dopamine hydrochloride, melamine, and cerium ammonium nitrate, and thoroughly soak them with anhydrous ethanol; S2: Transfer the mixture obtained in S1 to a polytetrafluoroethylene-lined hydrothermal reactor for reaction; S3: Centrifuge the solution from S2 after the reaction is complete at high speed, collect the supernatant and wash it with anhydrous ethanol; After washing, the solution was dialyzed with anhydrous ethanol. After dialysis, the solution in the dialysis bag was dried by rotary evaporation to obtain CeCDs solid, which was stored at -20°C.

2. The method for preparing a near-infrared absorbing cerium-doped carbon quantum dot biomimetic nanozyme according to claim 1, characterized in that: The ratio of dopamine hydrochloride, melamine, cerium ammonium nitrate, and anhydrous ethanol in S1 is 2 g: 2 g: 1 g: 300 mL.

3. The method for preparing a near-infrared absorbing cerium-doped carbon quantum dot biomimetic nanozyme according to claim 1, characterized in that: The reaction conditions in the polytetrafluoroethylene-lined hydrothermal reactor in S2 are 180°C for 12 hours.

4. The method for preparing a near-infrared absorbing cerium-doped carbon quantum dot biomimetic nanozyme according to claim 1, characterized in that: The high-speed centrifugation conditions in S3 are 20,000 rpm for 15 min. After centrifugation, the cells are washed twice. After washing, the dialysis bag used for dialysis has a retention mass of 1000 kDa.

5. A near-infrared absorbing cerium-doped carbon quantum dot biomimetic nanozyme, characterized in that: It is prepared using the preparation method described in any one of claims 1-4.

6. The near-infrared absorbing cerium-doped carbon quantum dot biomimetic nanozyme according to claim 5, characterized in that: The cerium-doped carbon quantum dot biomimetic nanozyme has a quantum dot structure with Ce-doped carbon dots as its core.

7. The near-infrared absorbing cerium-doped carbon quantum dot biomimetic nanozyme according to claim 5, characterized in that: The cerium-doped carbon quantum dot biomimetic nanozyme possesses nanozyme activity and can responsively generate hydrogen in response to 808nm near-infrared light.

8. The application of a near-infrared absorbing cerium-doped carbon quantum dot biomimetic nanozyme prepared by the preparation method described in claims 1-4 in the preparation of drugs for treating Parkinson's disease.

9. The application of the near-infrared absorbing cerium-doped carbon quantum dot biomimetic nanozyme according to claim 8 in the preparation of drugs for treating Parkinson's disease, characterized in that: The cerium-doped carbon quantum dot biomimetic nanozyme has SOD and CAT catalytic activities, which can reduce ROS generation.

10. The application of a near-infrared absorbing cerium-doped carbon quantum dot biomimetic nanozyme according to claim 8 in the preparation of drugs for treating Parkinson's disease, characterized in that: The nanozyme, as an SOD / CAT nanozyme, is used in the treatment of Parkinson's disease.

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