CN111569053A - Nano enzyme for enhancing immunotherapy by regulating and controlling tumor metabolic behaviors and preparation method and application thereof - Google Patents

Nano enzyme for enhancing immunotherapy by regulating and controlling tumor metabolic behaviors and preparation method and application thereof Download PDF

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CN111569053A
CN111569053A CN202010175171.3A CN202010175171A CN111569053A CN 111569053 A CN111569053 A CN 111569053A CN 202010175171 A CN202010175171 A CN 202010175171A CN 111569053 A CN111569053 A CN 111569053A
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oxidase
methylimidazole
nanoenzyme
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metal ions
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田华雨
张思佳
徐彩娜
林琳
陈杰
陈学思
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Changchun Institute of Applied Chemistry of CAS
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Abstract

The application belongs to the field of new biomedical materials, and discloses a nano enzyme for enhancing immunotherapy by regulating and controlling tumor metabolic behaviors, and a preparation method and application thereof. The nano enzyme is prepared from oxidase, metal ions, 2-methylimidazole and hyaluronic acid, and can enhance immunotherapy by regulating and controlling tumor metabolic behaviors and enhance the therapy of a PD-L1 antibody, so that the combination of hunger therapy, chemokinetic therapy and immunotherapy is realized, and a better treatment effect is achieved. The preparation method of the nano enzyme has the advantages of easy operation, low price and the like.

Description

Nano enzyme for enhancing immunotherapy by regulating and controlling tumor metabolic behaviors and preparation method and application thereof
Technical Field
The invention belongs to the field of new biomedical materials, and particularly relates to a nano enzyme for enhancing immunotherapy by regulating and controlling tumor metabolic behaviors, and a preparation method and application thereof.
Background
Cancer immunotherapy is to regulate the immune system of human body, utilize the immune cells of human body to resist tumor, and can induce long-term anti-tumor immune response. In 2018, nobel's physiology or medicine awarded two immunologists to show that they discovered cancer therapies that inhibit immune down-regulation. Currently the most clinically used immunotherapy is checkpoint therapy. However, most tumors respond less frequently to immune checkpoint therapy (Yarchoan M, Hopkins A, Jaffee EM. molar major and response rate to PD-1inhibition [ J ]. The New England and major of medicine,2017,377(25): 2500.). The most important factor affecting therapy is the immunosuppressive microenvironment. Therefore, the problem of tumor immune microenvironment inhibition is solved, and the cancer immunotherapy can exert the maximum effect.
Metabolism is the general term for a series of ordered chemical reactions that occur in the body of an organism to sustain life. In 1927, the nobel prize winner Otto Warburg observed that tumor cells consumed 200 times more glucose than normal cells, representing a significantly different metabolic phenotype; in 2010, Croig B.Thompson discovered the first tumor metabolite, 2-hydroxyglutarate (see Ward P S, Patel J, Wise D R, et al. the common feature of leucoma-associated IDH1 and IDH2 dynamics is a neoplasmic enzyme activity converting alpha-ketogluterate to 2-hydroxygluterate [ J ]. Cancer cell,2010,17(3): 225-). 234.). With the progress of research, researchers find that there are five metabolic pathways for tumor cells: aerobic glycolysis, glutaminolysis, one-carbon metabolism, pentose phosphate pathway and fatty acid de novo synthesis. These five metabolic pathways convert tumor cells from simple ATP production to the production of large amounts of amino acids, nucleotides, fatty acids, and other intermediates needed for rapid cell growth and proliferation.
In recent years, researches prove that metabolic disorder of tumor cells causes tumor immunosuppressive microenvironment, and metabolic disorder can inhibit immune cell infiltration, influence expression of cell surface markers and the like, thereby interfering immune monitoring. The metabolic pathways currently found to be associated with resistance to immunotherapy include hypoxia-inducible factors, adenosine, and the like. Therefore, the research of enhancing immunotherapy by regulating the metabolic behavior of tumors has huge development potential.
Disclosure of Invention
In view of the above, the present invention aims to provide a nano-enzyme for enhancing immunotherapy by regulating tumor metabolic behaviors, and a preparation method and an application thereof.
In order to achieve the purpose of the invention, the invention adopts the following technical scheme.
A nanometer enzyme is prepared from oxidase, metal ions, 2-methylimidazole and hyaluronic acid.
In the invention, the mass ratio of the oxidase, the metal ions and the 2-methylimidazole in the nanoenzyme is 1: (0.1-50): (0.01-30).
In the invention, the mass ratio of the oxidase/metal ions/2-methylimidazole to hyaluronic acid in the nanoenzyme is 1: (0.01-10).
In the invention, the oxidase in the nano-enzyme is one or two of glucose oxidase and lactate oxidase; the metal ion is Mn2+And Cu2+One or two of them.
In the invention, the molecular weight of the glucose oxidase in the nanoenzyme is 100000-160000 Da; the molecular weight of the lactate oxidase is 300000-350000 Da.
In the present invention, the Mn in the nanoenzyme2+Is manganese chloride tetrahydrate, the Cu2+Is cupric chloride dihydrate.
The invention also provides a preparation method of the nano enzyme, which is characterized by comprising the following steps:
A. dissolving oxidase and 2-methylimidazole in water for action, adding metal ions for mixing, centrifuging, removing supernate, and performing ultrasonic dispersion to obtain oxidase/metal ions/2-methylimidazole;
B. and uniformly mixing the hyaluronic acid aqueous solution with oxidase/metal ions/2-methylimidazole, and standing and compounding to obtain the nano enzyme.
According to the preparation method, firstly, oxidase and 2-methylimidazole act, then metal ions and 2-methylimidazole are added for coordination and blending to realize in-situ oxidase loading, and finally hyaluronic acid is used for modification to obtain oxidase/metal ions/2-methylimidazole/hyaluronic acid nanoenzyme.
In the present invention, the oxidase in step A of the preparation method is dissolved in water at a concentration of 1.5 mg/mL.
In the preparation method, the reaction time of the oxidase in the step A and 2-methylimidazole is 10-60 min. In some embodiments, the duration of action is 20 to 40 min.
In the preparation method, the metal ions are added in the step A for 0.5-2 h and then the mixture is centrifuged. In some embodiments, the centrifugation is performed 1 to 1.5 hours after the addition of the metal ions.
Further, the centrifugation speed is 8000-12000 rpm, and the centrifugation time is 10-30 min. In some embodiments, the centrifugation rate is 10000-12000 rpm, and the time is 20-30 min.
In the preparation method, the ultrasonic dispersion time in the step A is 0.5-15 min. In some embodiments, the ultrasonic dispersion time is 0.5 to 5 min.
In the present invention, in the preparation method, the hyaluronic acid aqueous solution in the step B is equal to the oxidase/metal ion/2-methylimidazole volume.
In the preparation method, the standing time in the step B is 10-60 min. In some embodiments, the standing time is 20 to 30 min.
The nano enzyme reaches a tumor region to release oxidase after being administered through tail vein, and the tumor metabolic behavior is regulated and controlled through a starvation therapy; secondly, the metal ions can generate Fenton-like reaction with hydrogen peroxide to generate active oxygen to kill tumors. Therefore, the invention also provides the application of the nano enzyme in preparing the medicine for regulating and controlling tumor metabolism and killing tumors.
In addition, the nano enzyme can enhance immunotherapy by regulating tumor metabolic behaviors and enhance the treatment of a PD-L1 antibody, so that the combination of hunger therapy, chemokinetic therapy and immunotherapy is realized, and a better treatment effect is achieved. Therefore, the invention also provides the application of the nano enzyme in the preparation of a PD-L1 antibody sensitizer.
According to the technical scheme, the invention provides the nano enzyme and the preparation method and the application thereof. The nano enzyme is prepared from oxidase, metal ions, 2-methylimidazole and hyaluronic acid, and can enhance immunotherapy by regulating and controlling tumor metabolic behaviors and enhance the therapy of a PD-L1 antibody, so that the combination of hunger therapy, chemokinetic therapy and immunotherapy is realized, and a better treatment effect is achieved. The preparation method of the nano enzyme has the advantages of easy operation, low price and the like.
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In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below.
FIG. 1 is a scanning electron micrograph of glucose oxidase/manganese ion/2-methylimidazole obtained in example 3 of the present invention;
FIG. 2 shows the cytotoxicity of nanoenzymes obtained in example 20 of the present invention;
FIG. 3 is a graph of the cellular level glucose consumption profile obtained in example 28 of the present invention; (1) a PBS group, (2) a glucose oxidase group, (3) a manganese ion/2-methylimidazole/hyaluronic acid group, (4) the nanoenzyme group obtained in example 20;
FIG. 4 is a graph showing reactive oxygen species staining at a cellular level obtained in example 29 of the present invention; (1) a PBS group, (2) a glucose oxidase group, (3) a manganese ion/2-methylimidazole/hyaluronic acid + hydrogen peroxide group, (4) the nanoenzyme group obtained in example 20;
FIG. 5 is a graph showing the results of detection of PD-L1 expression on tumor cells by flow cytometry, which was obtained in example 30 of the present invention; (1) PBS group, (2) manganese ion/2-methylimidazole group, (3) glucose oxidase/manganese ion/2-methylimidazole group, (4) nanoenzyme group obtained in example 20, (5) PD-L1 antibody group, (6) nanoenzyme + PD-L1 antibody group obtained in example 20.
Detailed Description
The invention discloses a nano enzyme for enhancing immunotherapy by regulating tumor metabolic behaviors and a preparation method and application thereof. Those skilled in the art can modify the process parameters appropriately to achieve the desired results with reference to the disclosure herein. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and products of the present invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications of the methods described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of the present invention without departing from the spirit and scope of the invention.
According to the invention, the nano enzyme consists of oxidase, metal ions, 2-methylimidazole and hyaluronic acid. Firstly, the oxidase and 2-methylimidazole act, then metal ions and 2-methylimidazole are added for coordination and blending to realize in-situ oxidase loading, and finally hyaluronic acid is used for modification to prepare the nano enzyme.
According to the invention, the molecular weight of the glucose oxidase is 100000-160000 Da. In some embodiments 140000 to 160000 Da. In some embodiments 150000 Da.
According to the invention, the molecular weight of the lactate oxidase is 300000-350000 Da. In some embodiments 330000 to 350000 Da. And in some embodiments 350000 Da.
The ratio of the oxidase to the metal ions to the 2-methylimidazole is 1: (0.1-50): (0.01-30). In some embodiments, 1: (0.5-20): (0.1-10). Further, in some embodiments, the compound is 1 (1-10): (0.6-4).
The ratio of oxidase/metal ions/2-methylimidazole to hyaluronic acid is 1: (0.01-10), in some embodiments 1: (0.01-3).
According to the invention, the preparation method of the nano enzyme comprises the following steps: dissolving oxidase and 2-methylimidazole in water, adding metal ion water solution to coordinate with 2-methylimidazole, and centrifuging. And (4) discarding the supernatant, and preparing oxidase/metal ions/2-methylimidazole after ultrasonic dispersion. Then evenly mixing the hyaluronic acid aqueous solution with oxidase/metal ions/2-methylimidazole, and statically compounding to obtain the oxidase/metal ions/2-methylimidazole/hyaluronic acid nanoenzyme.
According to the invention, the oxidase is reacted with 2-methylimidazole for a time of 10 to 60min, in some embodiments 20 to 40 min.
The coordination time of the added metal ions and the 2-methylimidazole is 0.5 to 2 hours, and in some embodiments 1 to 1.5 hours.
The centrifugation rate is 8000 to 12000rpm, in some embodiments 10000 to 12000 rpm; the centrifugation time is 10 to 30min, in some embodiments 20 to 30 min.
The ultrasonic dispersion time is 0.5 to 15min, in some embodiments 0.5 to 5 min.
The standing time is 10-60 min, in some embodiments 20-30 min.
According to the invention, the nano enzyme for enhancing immunotherapy by regulating tumor metabolic behavior releases oxidase after being administered through tail vein, and regulates tumor metabolic behavior by starvation therapy; secondly, the metal ions can generate Fenton-like reaction with hydrogen peroxide to generate active oxygen to kill tumors; in addition, the nano enzyme can enhance immunotherapy by regulating tumor metabolic behaviors and enhance the treatment of a PD-L1 antibody, so that the combination of hunger therapy, chemokinetic therapy and immunotherapy is realized, and a better treatment effect is achieved.
The experimental method related to each embodiment of the invention is carried out according to the following operations:
cell culture: in the present invention, the 4T1 cell line is selected, and the cell culture method is according to a general method without particular limitation. The culture medium is preferably DMEM medium containing 10% fetal bovine serum, and the culture conditions are preferably in an incubator with 5% volume carbon dioxide and 37 ℃.
Cytotoxicity evaluation 4T1 cell line was selected and cells were plated at 8 × 10 per well3Cultured in 96-well plates overnight. After co-culturing the material with different concentrations with cells for 24h, 20. mu.L of CCK-8 solution was added to each well and the culture was continued for 1 h. Absorbance values at 450nm and 610nm were measured for each well by a microplate reader. And the cell survival rate was calculated by the following formula.
Cell survival rate (%) ═ aSample (I)/ABlank space)×100
Cell-level glucose consumption ability evaluation of cell-level glucose consumption 4T1 cell line was selected and cells were plated at 1 × 10 per well5Cultured overnight in 24-well plates. After co-culturing the different materials with the cells for 24h, the sugar content in the medium in each well was determined with a glucometer.
Cellular level reactive oxygen species staining cell line 4T1 was selected for evaluation of cellular level reactive oxygen species staining cells were plated at 1 × 10 per well5Cultured overnight in 24-well plates. Different materials were co-cultured with the cells. 2',7' -dichlorofluoroxanthate was diluted with serum-free medium at a ratio of 1:1000 to a final concentration of 10. mu. mol/L. Removing the cell culture solution, adding diluted 2',7' -dichlorofluorescein diacetate, incubating at 37 deg.C for 20min, washing the cells with serum-free culture solution for three times, and detecting with 488nm excitation wavelength and 525nm emission wavelength under fluorescence microscope.
The combined treatment animal model is a 4T1 tumor model selected, and about 20g Balb/C mice selected, and the subcutaneous inoculation density of the right hind limb outside of the mice is 1 × 1064T1 cells, the tumor volume is 100mm3When it is left and right, willThe nano enzyme tail vein injection for enhancing the immunotherapy by regulating the tumor metabolic behavior is injected into a mouse body for 4 times, and the PD-L1 antibody abdominal cavity administration is carried out for 2 times in the combined treatment group. Tumor volume size was followed and PD-L1 expression on tumor cells was detected by flow cytometry at the end of the experiment.
In order to further understand the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Unless otherwise specified, the reagents involved in the examples of the present invention are all commercially available products, and all of them are commercially available.
Examples 1 to 16
Firstly, dissolving oxidase and 2-methylimidazole in water, acting for 30min, then adding a metal ion aqueous solution and 2-methylimidazole for coordination for 1h, and then centrifuging at the rotating speed of 12000rpm for 20 min. And (4) discarding the supernatant, and performing ultrasonic dispersion for 30s to prepare oxidase/metal ions/2-methylimidazole. Wherein, the dosage of the oxidase, the metal ion and the 2-methylimidazole is shown in the table 1.
TABLE 1 dosage of different raw materials of examples 1-16
Figure BDA0002410575870000071
Glucose oxidase/manganese ions/2-methylimidazole is prepared according to the dosage ratio of the embodiment 3, and the glucose oxidase/manganese ions/2-methylimidazole obtained in the embodiment 3 is analyzed by using a scanning electron microscope to obtain a scanning electron microscope picture, which is shown in fig. 1. The result shows that the average size of the glucose oxidase/manganese ions/2-methylimidazole prepared in the example 3 is 40-80 nm.
The particle sizes of examples 1-2 and 5-6 were smaller than those of example 3, and those of examples 4 and 7-16 were larger than those of example 3.
Examples 17 to 26
And (3) according to the embodiment 3, uniformly mixing the prepared glucose oxidase/manganese ions/2-methylimidazole with an equal volume of hyaluronic acid aqueous solution and the glucose oxidase/manganese ions/2-methylimidazole, and standing and compounding for 20min to obtain the glucose oxidase/manganese ions/2-methylimidazole/hyaluronic acid nanoenzyme. Wherein, the dosage of the glucose oxidase/manganese ion/2-methylimidazole and hyaluronic acid is shown in the table 2.
TABLE 2 dosage of different raw materials of examples 17 to 26
Figure BDA0002410575870000081
Nanoenzymes of each of examples 17-26 were prepared according to the above table amounts.
Example 27
4T1 cells were plated at 8 × 10 per well3Cultured in 96-well plates overnight. After co-culturing the nanoenzymes obtained in example 20 at different concentrations with cells for 24 hours, 20. mu.L of CCK-8 solution was added to each well, and the culture was continued for 1 hour. Absorbance values at 450nm and 610nm were measured for each well by a microplate reader. The cell viability was calculated and the results are shown in FIG. 2.
The experimental result shows that the nano-enzyme obtained in example 20 has a killing effect on 4T1 cells at a lower concentration.
The killing effect of the nanoenzymes obtained in examples 17-19 and 21-26 on 4T1 cells is similar to that of example 20.
Example 28
4T1 cells were plated at 1 × 10 per well5Was inoculated in 24-well plates and cultured overnight. The experiment was divided into four groups: (1) PBS group, (2) glucose oxidase group, (3) manganese ion/2-methylimidazole/hyaluronic acid group, (4) nanoenzyme group obtained in example 20, which was co-cultured with cells, respectively, (2) group and (4) group were added at a final glucose oxidase concentration of 2. mu.g/mL, (1) group was added with PBS of the same volume as the other group, (3) group was used in the same volume as manganese ion/2-methylimidazole/hyaluronic acid in group (4), and each group was measured with a glucometer after 24 hoursThe results of the sugar content in the well medium are shown in FIG. 3.
The results of the experiments showed that the glucose content in the medium was reduced after treatment with group (4) compared to groups (1) and (3), and the amount of the reduction was comparable to that of group (2). Therefore, the nanoenzyme obtained in example 20 has the ability to consume glucose.
The nanoenzymes obtained in examples 17 to 19 and 21 to 26 had glucose consuming abilities similar to those of example 20.
Example 29
4T1 cells were plated at 1 × 10 per well5The cells were co-cultured with the nanoenzyme groups obtained in example 20, (2) and (4) were added to the cells at a final glucose oxidase concentration of 5. mu.g/mL, (1) PBS was added to the group at an equal volume to the other groups, (3) the amount of the group was kept in accordance with the amount of manganese ions/2-methylimidazole/hyaluronic acid in the group (4) in terms of manganese ions/2-methylimidazole/hyaluronic acid, and (3) hydrogen peroxide in the group was used at 1 × 10-3Mu mol/L. According to the following steps of 1:1000 serum-free medium was used to dilute 2',7' -dichlorofluorescent yellow diacetate to a final concentration of 10. mu. mol/L. Removing the cell culture solution, adding diluted 2',7' -dichlorofluorescein diacetate, incubating at 37 deg.C for 20min, washing the cells with serum-free culture solution three times, using 488nm excitation wavelength and 525nm emission wavelength, and detecting with fluorescence microscope, the result is shown in FIG. 4.
The experimental results showed that (1) and (2) did not produce green fluorescence, i.e., no active oxygen was produced, while (3) and (4) produced a large amount of green fluorescence, i.e., a large amount of active oxygen was produced. Indicating Mn in the nanoenzyme obtained in example 202+Can generate fenton-like reaction with hydrogen peroxide generated by glucose oxidase consuming glucose to generate active oxygen to kill tumor.
Similar to example 20, the nanoenzymes obtained in examples 17-19 and 21-26 can also undergo Fenton-like reaction with hydrogen peroxide generated by glucose oxidase consuming glucose to generate active oxygen to kill tumors.
Example 30
Selecting about 20g of Balb/C mice, and dividing the mice into six groups, namely (1) a PBS group, (2) a manganese ion/2-methylimidazole group, (3) a glucose oxidase/manganese ion/2-methylimidazole group, (4) a nanoenzyme group obtained in example 20, (5) a PD-L1 antibody group and (6) a nanoenzyme + PD-L1 antibody group obtained in example 20, wherein 5 mice are inoculated at the outer side of the right hind limb of each mouse under the skin with the density of 1 × 1064T1 cells, the tumor volume is 100mm3On the left and right sides, the tail veins of each group were injected into mice for 4 times, each time with 2 days interval (if the first administration is recorded as D0, the last three times are D3, D6 and D9 respectively), the administration amount of each group was 20 μ g/body based on the final concentration of glucose oxidase, (2) the group dosage was kept consistent with the administration amount of manganese ions/2-methylimidazole in (4), (3) the group dosage was consistent with the administration amount of glucose oxidase/manganese ions/2-methylimidazole in (4), (5) and (6) the group were administered with PD-L1 antibody intraperitoneally 2 times (D4 and D8 respectively), and the administration amount was 50 μ g/body. Tumor volume size was followed and 14 days after the start of treatment the experiment was terminated and PD-L1 expression on tumor cells was detected by flow cytometry. The results are shown in Table 3 and FIG. 5.
TABLE 3 tumor volumes in each group
Grouping Tumor volume
(1) Group of 2000mm3
(2) Group of 1500mm3
(3) Group of 1000mm3
(4) Group of 500mm3
(5) Group of 1800mm3
(6) Group of 100mm3
The experimental results showed that group (6) completely inhibited tumor growth compared to the other control groups, so that a better therapeutic effect could be achieved by the combination of starvation therapy, chemokinetic therapy and immunotherapy.
The expression of PD-L1 on the tumor cells is detected by flow cytometry, and experimental results show that the expression of PD-L1 on the tumor cells of the (4) group and the (6) group is obviously up-regulated, so the nano enzyme obtained in example 20 can cause the expression of PD-L1 on the tumor cells to be up-regulated, and the PD-L1 antibody is sensitized for treatment, so that the immunotherapy can be enhanced by regulating and controlling the metabolic behavior of the tumor.
The nanoenzymes obtained in examples 17-19 and 21-26, similar to example 20, can inhibit tumor growth while enhancing immunotherapy by modulating tumor metabolic behavior.
The foregoing embodiments are merely provided to assist and understand the core concept of the method of the present invention, and it will be apparent to those skilled in the art that modifications and variations can be made in the embodiments without departing from the principles of the invention, and the modifications and variations are also considered to be within the scope of the invention.

Claims (10)

1. A nano enzyme is characterized by being prepared from oxidase, metal ions, 2-methylimidazole and hyaluronic acid.
2. The nanoenzyme of claim 1, wherein the mass ratio of the oxidase, metal ion and 2-methylimidazole is 1: (0.1-50): (0.01 to 30); the mass ratio of the oxidase/metal ions/2-methylimidazole to the hyaluronic acid is 1: (0.01-10).
3. The nanoenzyme of claim 1, wherein the oxidase is one or both of glucose oxidase and lactate oxidase; the metal ion is Mn2+And Cu2+One or two of them.
4. The nanoenzyme of claim 3, wherein the glucose oxidase has a molecular weight of 100000-160000 Da; the molecular weight of the lactate oxidase is 300000-350000 Da.
5. The nanoenzyme of claim 3, wherein the Mn is2+Is manganese chloride tetrahydrate, the Cu2+Is cupric chloride dihydrate.
6. The process for preparing nanoenzyme according to any one of claims 1 to 5, comprising the steps of:
A. dissolving oxidase and 2-methylimidazole in water for action, adding metal ions for mixing, centrifuging, removing supernate, and performing ultrasonic dispersion to obtain oxidase/metal ions/2-methylimidazole;
B. and uniformly mixing the hyaluronic acid aqueous solution with oxidase/metal ions/2-methylimidazole, and standing and compounding to obtain the nano enzyme.
7. The preparation method according to claim 6, wherein the reaction time of the oxidase in the step A and 2-methylimidazole is 10-60 min; centrifuging after adding the metal ions for 0.5-2 h; the centrifugation speed is 8000-12000 rpm, and the centrifugation time is 10-30 min; the ultrasonic dispersion time is 0.5-15 min.
8. The preparation method according to claim 6, wherein the standing time in the step B is 10-60 min.
9. Use of the nanoenzyme of any one of claims 1 to 5 in the preparation of a medicament for modulating tumor metabolism and killing tumors.
10. Use of the nanoenzyme of any one of claims 1 to 5 in the preparation of a PD-L1 antibody sensitizer.
CN202010175171.3A 2020-03-13 2020-03-13 Nano enzyme for enhancing immunotherapy by regulating and controlling tumor metabolic behaviors and preparation method and application thereof Pending CN111569053A (en)

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Application publication date: 20200825