CN115737788A

CN115737788A - Preparation and application of metal organic framework nano probe material for efficiently depriving glucose

Info

Publication number: CN115737788A
Application number: CN202211104632.3A
Authority: CN
Inventors: 曾雪梅
Original assignee: Fujian Normal University
Current assignee: Fujian Normal University
Priority date: 2022-09-09
Filing date: 2022-09-09
Publication date: 2023-03-07

Abstract

The invention discloses preparation and application of a metal organic framework nano probe material for efficiently depriving glucose, and belongs to the technical field of nano biological materials. According to the invention, zinc ions of zinc nitrate hexahydrate are taken as metal ions, 2-methylimidazole is taken as an organic framework, treatment preparations of mannose, glucose oxidase and peroxidase are added, and a metal organic framework nano probe material ZIF-8/M & G & H is synthesized by a one-pot method.

Description

Preparation and application of metal organic framework nano probe material for efficiently depriving glucose

Technical Field

The invention belongs to the technical field of nano biological materials, and particularly relates to preparation and application of a metal organic framework nano probe material for efficiently depriving glucose.

Background

Triple negative breast cancer is one of the most common malignant tumors of women, and has the problems of high heterogeneity, high malignancy, strong invasiveness, high recurrence rate, poor prognosis and the like, so that the treatment difficulty is high. The first choice for treating triple negative breast cancer is chemotherapy, but the traditional Chinese medicine has the defects of poor treatment effect, strong side effect, poor prognosis and the like. Local surgical excision also has the problems of high postoperative recurrence rate, high lymph node metastasis rate and the like. In addition, immune checkpoint therapy is only applicable to triple negative breast cancer patients with high expression of PD-L1 antibody, with certain limitations. Therefore, the development of an effective treatment scheme for triple negative breast cancer has very important scientific value and application value.

Tumor starvation therapy inhibits the growth of cancer cells by "starving" by limiting or depriving the growth of cancer cells of desirable conditions, such as pathways for angiogenesis, substance and energy metabolism. The hunger therapy has the advantages of rich treatment means, small toxic and side effects and the like, and is widely used for treating various cancers. Heretofore, various nanomaterials deprive cells of glucose through loading glucose oxidase, so that starvation treatment of tumors is realized. However, glucose oxidase-mediated starvation therapy has limited therapeutic efficacy and extremely high oxygen demand. Therefore, the active development of the combination of multiple treatment modes to improve the treatment effect of the hunger therapy has important scientific significance and practical significance.

Disclosure of Invention

Aiming at the defects in the prior art, the invention aims to provide preparation and application of a metal organic framework nano probe material for efficiently depriving glucose, the metal organic framework nano probe material, namely ZIF-8/M & G & H, which simultaneously loads zinc of various therapeutic agents is prepared by a one-pot method and is used for combined therapy of glucose oxidation and metabolic interference, so that the technical problem of poor effect of hunger therapy in the prior art is solved.

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

a preparation method of a metal organic framework nano probe material for efficiently depriving glucose comprises the following steps: zinc ions of zinc nitrate hexahydrate are used as metal ions, 2-methylimidazole is used as an organic framework, mannose, glucose oxidase and peroxidase serving as therapeutic agents are added, and the metal organic framework nano probe material ZIF-8/M & G & H is synthesized through a one-pot method.

Further, the preparation steps of the metal organic framework nano probe material ZIF-8/M & G & H are as follows:

1) Preparing mixed solution of zinc nitrate hexahydrate and therapeutic preparation

Dissolving zinc nitrate hexahydrate, mannose, glucose oxidase and peroxidase in water to obtain a mixed solution of the zinc nitrate hexahydrate and a therapeutic preparation;

2) Preparing 2-methylimidazole aqueous solution

Dissolving 2-methylimidazole in water to obtain a 2-methylimidazole water solution;

3) Preparation of ZIF-8/M & G & H

Under the condition of violent stirring, pouring a 2-methylimidazole water solution into a container, uniformly stirring, dropwise adding a mixed solution of zinc nitrate hexahydrate and the therapeutic preparation, continuously stirring at room temperature for 5-10 min, and then centrifugally washing away unreacted zinc ions, therapeutic preparation molecules and imidazole to obtain a precipitate, namely ZIF-8/M & G & H.

Preferably, the molar ratio of the zinc nitrate hexahydrate to the 2-methylimidazole is 1.

Preferably, the molar ratio of mannose to glucose oxidase to peroxidase is 180-220.

The addition proportion and the addition sequence of the therapeutic preparation can be adjusted.

The synthesized nano probe material ZIF-8/M & G & H can be used for preparing medicaments for treating tumors.

Furthermore, the medicine for treating the tumor is a medicine for treating the tumor by adopting the glucose oxidation and metabolic interference combined mode, and the tumor to be treated is breast cancer tumor.

Before the nanometer probe material ZIF-8/M & G & H is put into application, the nanometer probe material ZIF-8/M & G & H can be modified by polyethylene glycol so as to increase the water solubility and biocompatibility of the nanometer probe material. Specifically, the ZIF-8/M & G & H and DSPE-PEG are stirred and react at room temperature overnight to obtain the nano probe ZIF-8/M & G & H-PEG.

Generally, compared with the prior art, the technical scheme of the invention has the following beneficial effects:

(1) The invention can prepare the zinc metal organic framework nano material ZIF-8 and the nano probe material ZIF-8/M & G & H loaded with a therapeutic preparation by a one-pot method. The shape of ZIF-8 is not affected by the load of the therapeutic preparation, and the two materials are regular hexagons, have diameters of about 60-90 nm and have porous structures.

(2) The invention takes ZIF-8 as a carrier and ZIF-8 as MOF, and well utilizes the advantages of simple synthesis and permanent porous structure.

(4) The synthesized nano probe material ZIF-8/M & G & H takes ZIF-8 as a carrier, peroxidase and glucose oxidase form a cascade catalytic reaction, hydrogen peroxide in a tumor microenvironment can be catalyzed into oxygen when cancer treatment is carried out, the oxygen content in tumor cells is improved, and the problem of oxygen deficiency in the glucose oxidation process is further solved; mannose, an isomer of glucose, is capable of interfering with entry into cells by metabolism and with cellular glucose metabolism when used in cancer therapy, and its metabolites are incapable of performing the downstream tricarboxylic acid cycle. Therefore, the nano probe material ZIF-8/M & G & H can effectively reduce the glucose content of cells, and kill tumor cells by starvation.

Drawings

FIG. 1 is a schematic of a one-pot synthesis of ZIF-8/M & G & H.

FIG. 2 is a transmission electron micrograph of synthetic ZIF-8, ZIF-8/M & G & H, scale bars is 100 nm.

FIG. 3 is a blank panel, and the effect of ZIF-8 and ZIF-8/M & G & H on cell starvation therapy was evaluated after incubation with cells at scale bars of 200 μ M.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and do not limit the invention.

The one-pot method comprises the steps of mixing and stirring an aqueous solution of zinc nitrate hexahydrate and an aqueous solution of an organic frame to synthesize ZIF-8; or, mixing and stirring a mixed solution of mannose, glucose oxidase and peroxidase, an aqueous solution of zinc nitrate hexahydrate and an aqueous solution of an organic frame, and synthesizing ZIF-8/M & G & H by a one-pot method.

Furthermore, the ZIF-8/M & G & H is modified with PEG which can increase water solubility and biocompatibility to obtain the ZIF-8/M & G & H-PEG.

The nanometer probe materials ZIF-8/M & G & H and ZIF-8/M & G & H-PEG can catalyze hydrogen peroxide into oxygen, and when the probe is applied to cell therapy or living body therapy, the probe can catalyze the hydrogen peroxide enriched in cancer cells to be converted into oxygen, so that the requirement of a glucose oxidase catalysis process on oxygen is met.

Example 1

A preparation method of metal organic framework (ZIF-8/M & G & H) and ZIF-8/M & G & H-PEG (zinc indium gallium ethylene) nano-probe materials capable of efficiently depriving glucose comprises the following steps:

Dissolving 1-3mmol of zinc nitrate hexahydrate, 20 mg of mannose, 25 mu g of glucose oxidase and 25 mu g of peroxidase in 3mL of water to obtain a mixed solution of the zinc nitrate hexahydrate and a therapeutic preparation;

2) Preparing 2-methylimidazole aqueous solution

Dissolving 3-7g of 2-methylimidazole in 20mL of water to obtain a 2-methylimidazole water solution;

3) Preparation of ZIF-8/M & G & H

Pouring 2-methylimidazole water solution into a container under the condition of violent stirring, dropwise adding mixed solution of zinc nitrate hexahydrate and the therapeutic preparation after uniform stirring, continuously stirring for 5-10 min at room temperature until the color is changed from colorless to light milky color, and centrifugally washing away unreacted zinc ions, therapeutic preparation molecules and imidazole to obtain light white precipitate, namely ZIF-8/M & G & H;

4) Preparation of ZIF-8/M & G & H-PEG

Mixing DSPE-PEG (with the molecular weight of 5000 Da) and ZIF-8/M & G & H at room temperature overnight to obtain ZIF-8/M & G & H-PEG.

Example 2

A preparation method of metal organic framework (ZIF) -8/M & G & H and ZIF-8/M & G & H-PEG) nano probe materials for efficiently depriving glucose comprises the following steps:

Dissolving 15 mg of zinc nitrate hexahydrate, 20 mg of mannose, 25 mu g of glucose oxidase and 25 mu g of peroxidase in 0.5mL of ultrapure water, and uniformly dispersing by ultrasonic to obtain a mixed solution of the zinc nitrate hexahydrate and a therapeutic preparation;

2) Preparing 2-methylimidazole aqueous solution

Dissolving 33 mg of 2-methylimidazole in 1 mL of water, and uniformly dispersing by using ultrasonic waves to obtain a 2-methylimidazole water solution;

3) Preparation of ZIF-8/M & G & H

Under the condition of violent stirring, dropwise adding a mixed solution of zinc nitrate hexahydrate and a therapeutic preparation into 1 mL of 2-methylimidazole water solution, continuously stirring at room temperature for 10 min until the color is changed from colorless to light milky color, and centrifugally washing off unreacted zinc ions, therapeutic preparation molecules and imidazole to obtain milky-white precipitates, namely ZIF-8/M & G & H;

4) Preparation of ZIF-8/M & G & H-PEG

And mixing ZIF-8/M & G & H and a proper amount of DSPE-PEG, stirring overnight, centrifuging at 8000 rpm for 10 min, washing with ultrapure water for three times to remove unreacted PEG, and thus obtaining ZIF-8/M & G & H-PEG.

Example 3

This example provides methods for preparing ZIF-8 and ZIF-8/M & G & H

1) Preparation of ZIF-8

Under the condition of vigorous stirring, 0.5mL of solution in which 15 mg of zinc nitrate hexahydrate is dissolved is dropwise added into a water reaction system containing 1 mL of 33 mg of 2-methylimidazole water solution, the mixture is continuously stirred at room temperature for 10 min, the color is changed from colorless to light milky white, the mixture is centrifuged at 8000 rpm for 10 min, and unreacted metal ions and imidazole are washed away to obtain milky white precipitate, namely ZIF-8.

2) Preparation of ZIF-8/M & G & H

And (3) reacting a mixed solution of 15 mg of zinc nitrate hexahydrate, 20 mg of mannose, 25 mu G of glucose oxidase and 25 mu G of peroxidase with a 2-methylimidazole water solution to obtain ZIF-8/M & G & H.

As shown in fig. 2, the transmission electron microscope morphology of ZIF-8 and ZIF-8/M & G & H prepared in this example is shown, and both the two nanomaterials are regular hexagons, have diameters in the range of 60-90 nm, and have good dispersibility.

Application example 1

Organometallic framework materials for the evaluation of the effectiveness of in vitro treatment of breast cancer

And respectively incubating ZIF-8 and ZIF-8/M & G & H with mouse breast cancer cells (4T 1), and after 12H incubation, staining the cells with calcein and propidium iodide to characterize the influence of glucose deprivation of the ZIF-8/M & G & H on cell death and activity.

FIG. 3 is a result of cell confocal showing that the control group of cells without any treatment and the group of ZIF-8 cells were very little toxic to cells and almost all cells were stained with calcein, indicating that ZIF-8 did not cause cell death. The ZIF-8/M & G & H incubated cells showed abnormal cell morphology, and most of the cells were stained with propidium iodide, indicating that ZIF-8/M & G & H is able to efficiently deprive the cells of glucose, resulting in cell death.

It will be understood by those skilled in the art that the foregoing is only an exemplary embodiment of the present invention, and is not intended to limit the invention to the particular forms disclosed, since various modifications, substitutions and improvements within the spirit and scope of the invention are possible and within the scope of the appended claims.

Claims

1. A preparation method of a metal organic framework nano probe material capable of efficiently depriving glucose is characterized by comprising the following steps: zinc ions of zinc nitrate hexahydrate are used as metal ions, 2-methylimidazole is used as an organic framework, a therapeutic preparation is added, mannose, glucose oxidase and peroxidase are used as the therapeutic preparation, and the metal organic framework nano probe material ZIF-8/M & G & H is synthesized through a one-pot method.

2. The method for preparing the metal organic framework nano probe material for efficiently depriving glucose according to claim 1, which comprises the following steps:

2) Preparing 2-methylimidazole aqueous solution

3) Preparation of ZIF-8/M & G & H

3. The method for preparing a metal organic framework nano probe material capable of efficiently depriving glucose according to claim 2, wherein the molar ratio of zinc nitrate hexahydrate to 2-methylimidazole is 1.

4. The method for preparing the metal-organic framework nanoprobe material capable of efficiently depriving glucose according to claim 2, wherein the molar ratio of mannose, glucose oxidase and peroxidase is 180-220.

5. The metal organic framework nano probe material obtained by the preparation method according to any one of claims 1 to 4.

6. The use of the metal organic framework nanoprobe material of claim 5 in the preparation of a medicament for the treatment of a tumor.

7. The application of claim 6, wherein the ZIF-8/M & G & H is modified with polyethylene glycol to obtain the ZIF-8/M & G & H-PEG nanoprobe material before application.

8. The use of claim 6, wherein the medicament for treating tumor is a medicament for treating tumor by combination of glucose oxide and metabolic disturbance, and the tumor to be treated is breast cancer tumor.