CN114767711A

CN114767711A - Efficient redox nano enzyme for tumor treatment and preparation method thereof

Info

Publication number: CN114767711A
Application number: CN202210541797.0A
Authority: CN
Inventors: 徐铸巍; 应刚
Original assignee: Shenzhen Youse Biotechnology Co ltd
Current assignee: Enqi Biotechnology Shanghai Co ltd
Priority date: 2022-05-17
Filing date: 2022-05-17
Publication date: 2022-07-22
Anticipated expiration: 2042-05-17
Also published as: CN114767711B

Abstract

The invention discloses a high-efficiency redox nano enzyme for treating tumors and a preparation method thereof. The invention utilizes defect engineering strategy to develop simple and effective redox nano-enzyme by constructing enzyme simulation active center, and part of tungsten oxide is replaced by iron to form iron tungsten oxide, and the process can activate structure reconstruction and generate a large amount of defect sites including iron substitution and oxygen vacancy defects, which obviously improves binding capacity and catalytic activity. Under the irradiation of near-infrared laser, the designed iron-tungsten oxide nanoenzyme can simulate the cascade reaction of catalase and redox active enzyme, induce the substantial destruction of the redox and metabolic homeostasis of a tumor area, and thus remarkably enhance the tumor treatment effect. The invention provides a new method for developing high-efficiency redox nano enzyme for treating tumors.

Description

Efficient redox nano enzyme for tumor treatment and preparation method thereof

Technical Field

The invention belongs to the field of nano material preparation and biomedicine, and particularly relates to a high-efficiency redox nano enzyme for treating tumors and a preparation method thereof.

Background

More and more attention is focused on nanoenzyme engineered nanomaterials with enzyme mimetic properties, as they can provide a new platform for developing more advanced tumor-specific therapeutic strategies in biological systems, in particular metal nanoenzymes comprising redox activity, called redox nanoenzymes. It is highly sought after because of its inherent redox properties, which can trigger catalytic reactions in situ and break the relatively fragile tumor homeostasis. However, the development of highly efficient redox nanoenzymes for cancer therapy, especially those similar to native enzymes, remains a great challenge.

The lack of catalytically active centers is the biggest obstacle facing redox nanoenzymes. Currently, research is mainly to introduce catalytically active biomacromolecule peptides, aptamers, antibodies and the like into a nanoenzyme framework to endow the nanoenzyme framework with a substrate binding function, or to manipulate the metal center inside the nanoenzyme and the surrounding atomic configuration, so as to generate a basic catalytically active site. Nevertheless, the problem is that the complex design and synthesis processes limit their potential. More importantly, these approaches have over-focused on a single active moiety involved in catalysis, largely ignoring the synergistic mechanism between different active moieties in the native enzyme. The superior functions and activities achieved by native enzymes are closely related to the cooperation between the internal active sites, and the two key components of native enzymes, the substrate binding region and the catalytic site, even though they may be far apart in the primary structure, can still form a spatial conformation by folding and winding of the peptide chain and work together at the active center. Therefore, the key step in achieving a true oxidoreductase mimetic is to design the accompanying binding region and the co-catalytic site in a fixed space, and there is currently no related nanoenzyme design.

Technical problem to be solved

Aiming at the challenges in the prior art, the embodiment of the invention aims to provide a high-efficiency redox nanoenzyme for tumor treatment and a preparation method thereof.

(II) technical scheme

In order to achieve the purpose, the invention mainly adopts the following technical scheme,

the invention firstly discloses a novel redox nano-enzyme Fe-WOv developed through defect engineering, which is characterized in that: the nano material is WOx nano particles, wherein part of the WOx nano particles are substituted by Fe to form Fe-WOv nano enzyme. The preparation method comprises the following steps:

the first step is as follows: mixing Tannic Acid (TA), Na₂WO₄·7H₂O and FeCl₃Dissolved in 35mL of distilled water. Stirring vigorously for 20-60 minutes, and forming a W/Fe-TA complex by self-assembly of metal ions and tannic acid in an aqueous solution based on strong interaction of the metal ions and Tannic Acid (TA) catechol groups;

preferably, the mass of TA added is 0.6g

Preferably, said addition of Na₂WO₄Has a mass of 0.43g

Preferably, said addition of FeCl₃Has a mass of 0.79g

The second step is that: transferring the mixed solution into a 50mL polytetrafluoroethylene autoclave, and reacting for 5-20 hours at 120-200 ℃. The Fe-WOv nanostructure is synthesized by carrying out oxidative self-polymerization on an organic ligand in a hydrothermal process and carrying out hydrothermal treatment on a metal-tannin complex by utilizing a metal polyphenol coordination strategy. Finally, the product was collected and washed 3 times with distilled water.

WOx NPs are obtained by the above synthesis method without adding FeCl₃。

The invention further discloses application of the novel redox nano enzyme Fe-WOv nano material, namely, the redox and metabolic steady state substantial destruction of a tumor region can be induced through a mimic enzyme cascade reaction, so that the tumor treatment effect is obviously enhanced.

(III) advantageous technical effects

Compared with the prior art, the invention has the following advantages:

1. the invention develops a novel redox nano enzyme Fe-WOv through defect engineering. Fe doping in WOx (Fe-WOv) can induce simultaneous massive Fe substitution and Oxygen Vacancy (OV) defect formation, while these defective sites coordinate and exert the binding and catalytic functions observed in most native enzymes, thereby significantly enhancing Fe-WO_vBinding capacity and catalytic activity of nano enzyme.

2. Under the irradiation of NIR-II laser, the novel redox nanoenzyme Fe-WOv prepared by the invention can induce the redox and metabolic homeostasis in a tumor area to generate substantial damage through enzyme-like cascade reaction, thereby obviously improving the tumor treatment effect.

Drawings

FIG. 1 is a Transmission Electron Microscope (TEM) morphology of Fe-WOv nanoenzyme

FIG. 2 is a particle size distribution diagram of Fe-WOv nanoenzyme

FIG. 3 is a graph showing the decomposition performance of hydrogen peroxide

FIG. 4 is a graph showing glutathione consumption performance

Detailed Description

Example 1

Mixing 0.6g TA, 0.43g Na₂WO₄·7H₂O and 0.79g FeCl₃Dissolved in 35mL of distilled water. After stirring vigorously for 30 minutes, the mixed solution was transferred to a 50mL polytetrafluoroethylene autoclave and reacted at 160 ℃ for 10 hours. And finally, collecting the product, washing the product for 3 times by using distilled water, and synthesizing to obtain the Fe-WOv nano enzyme. WOx NPs are obtained by the above synthesis method without adding FeCl₃. The morphology of the Fe-WOv nanoenzyme structure was examined using a Transmission Electron Microscope (TEM), and as a result, as shown in FIG. 1, the obtained Fe-WOv nanoenzyme was composed of rosette two-dimensional nanosheets with sharp edges, and the average diameter thereof was about 96nm, which is very consistent with the results observed in Dynamic Light Scattering (DLS) in FIG. 2.

Example 2

Titanium sulfate (Ti (SO) is used₄)₂) Colorimetric detection of hydrogen peroxide decompositionAnd (4) performance. 0.5mL hydrogen peroxide (2.5 mL mM) and 0.5mL PBS, WOx or Fe-WOv nanoenzyme solution for 30 minutes S3 stage the mixed solution was added to Ti (SO)₄)₂To the solution, mix 1.33 preparation 24% Ti (SO) was added to 50mL of distilled water₄)₂And 8.33mL H₂SO₄. After ten minutes of addition of hydrogen peroxide, the concentration of hydrogen peroxide was determined by measuring its absorbance at 405 nm at room temperature. The continuous catalytic effect was verified by repeated addition of 2.5mm of hydrogen peroxide in solution. H was determined by the same method₂O₂The residual amount of (2) was the same as above. The hydrogen peroxide decomposition performance test result is shown in figure 3, and the result shows that H is obtained after the Fe-WOv nano enzyme solution is added₂O₂The increase in the concentration of dissolved oxygen in the treatment solution clearly determines that the CAT-like enzyme activity is high. In contrast, negligible activity was observed in WOx treatment.

Example 3

Glutathione (GSH) is another important intracellular metabolite that can scavenge OH in tumor cells and enhance the resistance of tumor cells to oxidative stress. Most metal-based catalysts neutralize rapidly when exposed to GSH, thus undoubtedly lowering the therapeutic index. Glutathione consumption assay procedure was as follows, DTNB solution (3.0 mg. multidot.mL)^-1100. mu.L) and GSH solution (1mM, 200. mu.L) were added Fe-WOv nanoenzyme solution (20, 40, 60, 80, 100. mu.g.mL)^-1) In (1). The mixture was then kept at 25 ℃ for 30 minutes under magnetic stirring, and the residual amount of GSH was determined by measuring the absorbance at 412 nm. The background solution is Fe-WOv nano enzyme solution. The consumption detection result of glutathione is shown in figure 4, and the result shows that the Fe-WOv nano enzyme has good enzyme capability similar to redox activity, can effectively consume GSH, has the GSH conversion rate reaching 80 percent, is obviously superior to WOx nano enzyme, and can be more efficiently used for treating tumors.

Claims

1. A high-efficiency redox nano-enzyme for treating tumors is characterized in that: the nano material is tungsten oxide (WOx) nano particles, wherein part of the nano particles are replaced by iron to form iron tungsten oxide (Fe-WOv) nano enzyme.

2. A method for preparing the nanomaterial of claim 1, comprising the steps of: the first step is as follows: mixing Tannic Acid (TA), Na₂WO₄7H2O and FeCl₃Dissolved in 35mL of distilled water.

Stirring vigorously for 20-60 minutes, and forming a W/Fe-TA complex by self-assembly of metal ions and tannic acid in an aqueous solution based on strong interaction of the metal ions and Tannic Acid (TA) catechol groups.

The second step is that: and transferring the mixed solution into a 50mL polytetrafluoroethylene autoclave, and reacting for 5-20 hours at 120-200 ℃. Organic ligands are oxidized and polymerized by self in a hydrothermal process, and a metal-tannin complex is hydrothermally treated by utilizing a metal polyphenol coordination strategy to synthesize the Fe-WOv nano structure. Finally, the product was collected and washed 3 times with distilled water.

3. The production method according to claim 2, characterized in that: adding 0.6g of TA and Na₂WO₄Has a mass of 0.43g, and the added FeCl₃The mass of (2) was 0.79 g.

4. A highly efficient redox nanoenzyme according to claim 1.

5. The use of the efficient redox nanoenzyme of claim 5, wherein: the Fe-WOv nanoenzyme can simulate cascade reaction of catalase and redox active enzyme, induce redox and metabolic homeostasis of tumor region to substantially destroy, thereby significantly enhancing tumor treatment effect.