Nanoreactor for tumor chemodynamic therapy and preparation method and application thereof
By designing the nanoreactor Cu-ZIF8@GOX@HAase, the problem of low ROS generation and drug delivery efficiency in CDT was solved by using porous metal organic framework materials and enzyme-loaded nanoreactors, and the continuous production of ROS and significant inhibition of tumor growth was achieved.
Patent Information
- Application Number
- CN202510118008.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-24
- Publication Date
- 2025-05-30
AI Technical Summary
In tumor chemodynamic therapy (CDT), the low concentration of endogenous H2O2 in the tumor microenvironment, the strong antioxidant defense of tumor cells, and the physical barrier formed by extracellular matrix limit the production of ROS and the delivery efficiency of CDT drugs.
A nanoreactor Cu-ZIF8@GOX@HAase was designed to promote ROS production and tumor permeability enhancement through the porous metal-organic framework material Cu-ZIF8 as a carrier, loading glucose oxidase (GOX) and hyaluronidase (HAase) under acidic conditions.
The continuous production of ROS is achieved, the supply of oxygen and glucose in the tumor is enhanced, the tumor growth is significantly inhibited, and the efficiency of CDT is improved.
Smart Images
Figure CN120053683A_ABST
Abstract
Description
Technical Field
[0001] This application relates to the field of pharmaceutical technology, and particularly to a nano-reactor for tumor chemodynamic therapy, its preparation method and application. Background Art
[0002] Chemodynamic therapy (CDT) is an emerging cancer treatment strategy that utilizes hydrogen peroxide (H 2 O 2 ) in the tumor microenvironment (TME) to be converted into a large amount of reactive oxygen species (ROS), especially hydroxyl radicals (·OH), through the Fenton reaction to destroy the structure and function of cancer cells. Compared with traditional surgical, radiotherapy and chemotherapy methods, CDT has significant advantages such as high selectivity, low drug resistance, high versatility, and small side effects.
[0003] However, CDT has certain limitations. For example, the low endogenous H 2 O 2 concentration in the tumor microenvironment, the strong antioxidant defense of tumor cells, and the physical barrier formed by the extracellular matrix (ECM). These factors limit the generation of ROS and the delivery efficiency of CDT drugs. ECM mainly contains collagen, elastin and hyaluronic acid, which are the main reasons for the hindered drug diffusion and increased interstitial pressure. Moreover, the presence of ECM limits the supply of glucose in the tumor, which leads to a reduction in the continuous generation of ROS and a weakened antitumor effect. Summary of the Invention
[0004] Related technologies have proposed a nano-reagent containing glucose oxidase (GOX) and Fenton reagent, and another related technology has proposed a polymer-encapsulated hyaluronidase (HAase) nanozyme.
[0005] For the nano-reagent containing GOX and Fenton reagent, it can consume glucose in the tumor to be converted into H 2 O 2 and generate ROS, thereby enhancing the CDT effect. However, this scheme is affected by the limitation of glucose supply inside the tumor and cannot achieve the continuous generation of ROS.
[0006] For the polymer-encapsulated HAase nanozyme, it can effectively degrade hyaluronic acid in the tumor ECM. By degrading the ECM, it can increase the concentration of nano-drugs and small molecules in the tumor, including glucose. Although these works have highlighted the potential of ECM degradation, they have not combined ECM degradation with GOX and Fenton reactions to maintain the generation of ROS.
[0007] To solve at least one of the above technical problems, the present application proposes a nano-reactor Cu-ZIF8@GOX@HAase (CZGH) for tumor chemodynamic therapy, its preparation method and application.
[0008] In a first aspect, there is provided a nano-reactor for tumor chemodynamic therapy, comprising a porous metal-organic framework material (Cu-ZIF8) and glucose oxidase (GOX) and hyaluronidase (Haase) loaded on the Cu-ZIF8.
[0009] In a second aspect, a preparation method of the nano-reactor as described in the first aspect is proposed, comprising:
[0010] Dissolve dimethylimidazole in deionized water to obtain a dimethylimidazole solution;
[0011] Disperse GOX and HAase in the dimethylimidazole solution, then add zinc acetate and copper nitrate, and stir to obtain an intermediate solution;
[0012] Centrifuge the intermediate solution and collect the resulting precipitate;
[0013] Purify the precipitate to obtain the nano-reactor.
[0014] In some possible embodiments, the step of dissolving dimethylimidazole in deionized water to obtain a dimethylimidazole solution comprises:
[0015] Add 65.68 parts by mass of dimethylimidazole to 10 parts by volume of deionized water at room temperature, and stir to obtain the dimethylimidazole solution, wherein the ratio of the parts by mass to the parts by volume is mg / ml.
[0016] In some possible embodiments, the step of dispersing GOX and HAase in the dimethylimidazole solution, then adding zinc acetate and copper nitrate, and stirring to obtain an intermediate solution comprises:
[0017] Disperse 0.25 parts by mass of GOX and 0.25 parts by mass of HAase in 10 parts by volume of the dimethylimidazole solution, then add 36.24 parts by mass of zinc acetate and 12.08 parts by mass of copper nitrate, and stir well at room temperature until completely dissolved to obtain the intermediate solution.
[0018] In some possible embodiments, the step of centrifuging the intermediate solution and collecting the resulting precipitate comprises:
[0019] Centrifuge the intermediate solution at a speed of 1000 rpm for 15 min and collect the resulting precipitate.
[0020] In some possible embodiments, the purification of the precipitate includes:
[0021] Ultrafiltration centrifugal filtration of the precipitate using an ultrafiltration tube.
[0022] In a third aspect, there is provided an application of the nano-reactor as described in the first aspect in the preparation of a drug for treating tumors.
[0023] According to the nano-reactor provided by the present application, using a porous metal-organic framework material (Cu-ZIF8) as a carrier, and simultaneously delivering glucose oxidase and hyaluronidase, it can release GOX and HAase under acidic conditions, utilize HAase to degrade the extracellular matrix, enhance tumor permeability, promote glucose entry into the tumor, ensure that the ROS cycle generator system obtains sufficient glucose supply from the blood, and utilize the H2O2 generated by GOX to oxidize and hydrolyze the glucose entering the tumor to generate ROS, forming a sustainable ROS cycle generator. This strategy that integrates ECM degradation and ROS generation provides a new solution for improving the efficiency and effect of CDT. Description of the Drawings
[0024] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the accompanying drawings of the embodiments will be briefly introduced below. Obviously, the accompanying drawings in the following description only relate to some embodiments of the present application and do not limit the present application.
[0025] Figure 1 It is a flowchart of the preparation method of the nano-reactor provided by the embodiment of the present application.
[0026] Figure 2 It is the elemental imaging result of the nano-reactor prepared in the embodiment of the present application.
[0027] Figure 3 It is the trend of the particle size change of the nano-reactor prepared in the embodiment of the present application when placed in different solutions for different times.
[0028] Figure 4 It shows the particle size change of the nano-reactor prepared in the embodiment of the present application in solutions with different pH values. Detailed Embodiments
[0029] To make the objectives, technical solutions, and advantages of this application clearer, the technical solutions of the embodiments of this application will be clearly and completely described below in conjunction with the accompanying drawings of the embodiments of this application. Obviously, the described embodiments are some, but not all, of the embodiments of this application. All other embodiments obtained by those of ordinary skill in the art based on the described embodiments of this application without creative efforts fall within the scope of protection of this application. It can be understood that, without conflict, some technical means described in the various embodiments herein may be replaced or combined with each other.
[0030] In the description of this application, references to "one embodiment" or "some embodiments" etc. mean that a particular feature, structure, or characteristic described in conjunction with the embodiment is included in one or more embodiments of this application. Thus, the phrases "in one embodiment", "in some embodiments", "in other some embodiments", "in still other embodiments", etc. that appear in different places in this specification are not necessarily all referring to the same embodiment, but mean "one or more but not all embodiments", unless otherwise specifically emphasized in another way.
[0031] Figure 1 The preparation method of the nano-reactor Cu-ZIF8@GOX@HAase (CZGH) provided by one embodiment of this application is shown, and this method includes the following steps ① to ④:
[0032] ① Add 65.68 mg of dimethylimidazole to 10 ml of room-temperature deionized water and stir to obtain a dimethylimidazole solution.
[0033] ② Disperse 0.25 mg of glucose oxidase (GOX) and 0.25 mg of hyaluronidase (HAase) in the 10 ml of dimethylimidazole solution obtained in step ①, then add 36.24 mg of zinc acetate and 12.08 mg of copper nitrate, and stir well at room temperature for 10 s to completely dissolve them to obtain an intermediate solution.
[0034] ③ Centrifuge the second solution obtained in step ② at a rotation speed of 1000 rpm for 15 min, and collect the resulting precipitate.
[0035] ④ Use an ultrafiltration tube to perform ultrafiltration centrifugation filtration (an example of a purification method) on the precipitate obtained in step ③ to obtain the nano-reactor as a blue product.
[0036] The nano-reactor Cu-ZIF8@GOX@HAase prepared according to the method of this embodiment, wherein GOX can oxidize glucose into gluconic acid and hydrogen peroxide. Due to the formation of gluconic acid, the activity of GOX can lower the pH value of the surrounding environment, while H 2 O 2is an important reactive oxygen species and can be converted into a large amount of ROS through the Fenton reaction. HAase can degrade hyaluronic acid in the ECM and increase the concentration of nanodrugs and glucose in tumors. Porous metal-organic framework material (Cu-ZIF8) is a metal-organic framework (MOF) material formed by the self-assembly of copper ions and 2-methylimidazole. Due to the presence of copper ions, Cu-ZIF8 has good catalytic activity. The products of GOX and HAase hydrolyzing glucose and hyaluronic acid can react with Cu ions in Cu-ZIF8 to generate a high concentration of ROS, promoting tumor cell death. Moreover, the pore structure of Cu-ZIF-8 also enables it to be used as a drug carrier for controlled drug release. In addition, Cu-ZIF8 has good stability and biocompatibility, and using it as a carrier can reduce the side effects of the drug itself on the human body. Cu-ZIF8 will decompose in an acidic environment, which coincides with the weak acidic environment of the tumor microenvironment. Loading GOX and HAase into it can endow it with the function of passive targeting.
[0037] Based on this, the nanoreactor prepared by the method according to this embodiment, using Cu-ZIF8 as a carrier and delivering glucose oxidase and hyaluronidase simultaneously, can release GOX and HAase under acidic conditions, utilize HAase to degrade the extracellular matrix, enhance tumor permeability, promote glucose to enter the tumor, ensure that the ROS cycle generator system obtains sufficient glucose supply from the blood, and utilize the H2O2 generated by GOX to oxidize and hydrolyze the glucose entering the tumor to generate ROS, forming a sustainable ROS cycle generator, which helps to improve the efficiency of tumor CDT.
[0038] The technology of this application has been verified to be feasible through experiments. In a breast cancer mouse model, CZGH shows the potential of CDT in both in vitro and in vivo studies. In addition, in a 4T1 tumor mouse model, CZGH hydrolyzes the ECM of solid tumors, increases the supply of oxygen and glucose, and significantly inhibits tumor growth. High-resolution photoacoustic (PA) functional imaging confirms that CZGH can increase tumor oxygen saturation. These results indicate that CZGH represents a new method for preparing CDT reagents and has good prospects in efficient tumor CDT through continuous ROS generation and enhanced tumor penetrability.
[0039] Figure 2 This is the elemental imaging result of the nanoreactor (nanoparticle) prepared in the embodiment of this application. It can be seen from this figure that the nanoparticle contains Cu and Zn elements, proving successful synthesis. In the upper left picture of this figure, the horizontal bright line in the lower right part of it represents a scale of 100 nm.
[0040] Figure 3The particle size change trend of the nano-reactor prepared in the embodiment of the present application when placed in different solutions for different times can be seen from this figure, indicating that the synthesized nano-reactor has good solution stability.
[0041] Figure 4 The particle size change of the nano-reactor prepared in the embodiment of the present application in solutions with different pH values is shown. It can be seen from this figure that the particle size of the synthesized nano-reactor decreases significantly under acidic conditions, mainly because ZIF-8 degrades under acidic conditions.
[0042] According to the previous introduction, it can be known that the nano-reactor Cu-ZIF8@GOX@HAase synthesized by the preparation method according to the above embodiment of the present application can be used for tumor chemodynamic therapy. Based on this, the nano-reactor can be applied to the preparation of drugs for treating tumors.
Claims
1. A nanoreactor for tumor chemodynamic therapy, characterized in that: The invention comprises a porous metal organic framework material (Cu-ZIF8) and glucose oxidase (GOX) and hyaluronidase (Haase) loaded on the Cu-ZIF8.
2. A method for preparing a nanoreactor as claimed in claim 1, characterized in that: include: Dissolving dimethylimidazole in deionized water to obtain a dimethylimidazole solution; Dispersing GOX and HAase in the dimethylimidazole solution, adding zinc acetate and copper nitrate, and stirring to obtain an intermediate solution; Centrifuging the intermediate solution and collecting the resulting precipitate; The precipitate is purified to obtain the nanoreactor as a blue product.
3. The preparation method according to claim 2, characterized in that: The step of dissolving dimethylimidazole in deionized water to obtain a dimethylimidazole solution comprises: 65.68 parts by mass of dimethylimidazole are added to 10 parts by volume of room temperature deionized water, and stirred to obtain the dimethylimidazole solution, wherein the ratio of the parts by mass to the parts by volume is mg / ml.
4. The preparation method according to claim 3, characterized in that: The GOX and HAase are dispersed in the dimethylimidazole solution, and then zinc acetate and copper nitrate are added and stirred to obtain an intermediate solution, including: 0.25 parts by mass of GOX and 0.25 parts by mass of HAase were dispersed in 10 parts by volume of the dimethylimidazole solution, and 36.24 parts by mass of zinc acetate and 12.08 parts by mass of copper nitrate were added, and stirred at room temperature to completely dissolve them, thereby obtaining the intermediate solution.
5. The preparation method according to any one of claims 2 to 4, characterized in that: The step of centrifuging the intermediate solution and collecting the resulting precipitate comprises: The intermediate solution was centrifuged at 1000 rpm for 15 min, and the resulting precipitate was collected.
6. The preparation method according to claim 1, characterized in that: The step of purifying the precipitate comprises: The precipitate is subjected to ultrafiltration centrifugal filtration using an ultrafiltration tube.
7. Use of the nanoreactor as claimed in claim 1 in preparing drugs for treating tumors.