CN113181358A - Application of Fe-N-C monatomic nanoenzyme - Google Patents
Application of Fe-N-C monatomic nanoenzyme Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0052—Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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Abstract
The invention relates to the field of medicines, in particular to application of Fe-N-C monatomic nanoenzyme. The invention provides a new application of Fe-N-C monatomic nanoenzyme, namely the application of Fe-N-C monatomic nanoenzyme in preparing antitumor drugs. The experimental result shows that the Fe-N-C monatomic nanoenzyme can catalyze H2O2Formation of OH‑Under the irradiation of 808nm ultraviolet laser, the light energy can be converted into heat energy, and the double-function killing device has the double-function of killing tumor cells. The invention provides a new treatment scheme for tumor treatment and has wide application prospect.
Description
Technical Field
The invention relates to the field of medicines, in particular to application of Fe-N-C monatomic nanoenzyme.
Background
In living cells, various enzyme-linked reactions are involved, Peroxidase (POD) participates in various key oxidation reactions, and the survival of cells is determined by reactive oxygen Radicals (ROS) formed, which are broadly referred to as free radicals and non-free radicals of oxygen origin, and contain superoxide anions (O)2 -) Hydrogen peroxide (H)2O2) Hydroxyl radical (OH)-) Ozone (O)3) And singlet oxygen: (1O2) Cells tend to maintain redox homeostasis. However, tumor cells generally exhibit an oxidative stress state (an imbalance between ROS production and antioxidant defense, tending to oxidize), so that the concentration of ROS in tumor cells is oftenCompared with normal cells, the tumor cells collect a large amount of H in comparison with the normal cells by 100 times2O2H is treated with exogenous peroxidase or with enhanced expression of peroxidase2O2Conversion to OH-Can promote the death of tumor cells and achieve the effect of resisting tumors. The monatomic catalyst has wide prospect in the field of molecular catalysis.
Currently, photothermal therapy (PTT) is receiving increasing attention as an emerging therapy in cancer treatment, in addition to immunotherapy and gene therapy. PTT takes advantage of the photothermal effect of photothermal conversion agents (PTAs), extracts energy from light and converts light energy into thermal energy to increase the temperature of the surrounding environment and initiate death of cancer cells. Noble metal materials with strong oxidation resistance are one of the most studied inorganic PTAs. Noble metal PTAs, including Au, Ag, Pt, and Pd, absorb laser light to excite electrons from a ground state to an excited state, and then release energy in the form of heat by non-radiative decay. The limitations of noble metal-based PTAs in PTT have prompted scientists to seek other inorganic or carbon-based materials as PTAs.
In view of the above, the current antitumor treatment methods are limited, and the development of novel antitumor drugs is urgently needed.
Disclosure of Invention
In view of the above, the technical problem to be solved by the present invention is to provide an application of Fe-N-C monatomic nanoenzyme, in particular, an application of Fe-N-C monatomic nanoenzyme in the preparation of antitumor drugs.
The invention provides application of Fe-N-C monatomic nanoenzyme in preparation of antitumor drugs.
The invention also provides application of the Fe-N-C monatomic nanoenzyme as peroxidase.
The invention also provides a Fe-N-C monatomic nanoenzyme for preparing and reducing H in tumor cells2O2Application in medicine.
The invention also provides application of the Fe-N-C monoatomic nanoenzyme as a photothermal conversion agent.
The invention also provides application of the Fe-N-C monatomic nanoenzyme in preparation of anti-breast cancer drugs.
Preferably, the method comprises the following steps: free radicals generated by Fe-N-C monatomic nanoenzyme through photothermal action and Fenton reaction kill breast cancer cells.
Preferably, the laser wavelength used for the photothermal action is 808 nm;
the irradiation power of the laser is 1.0W cm-2The irradiation time was 10 min.
Preferably, the dosage of the Fe-N-C monatomic nanoenzyme is 3.125 mu g/mL-100 mg/mL.
Preferably, the anti-breast cancer drug comprises Fe-N-C monatomic nanoenzyme and pharmaceutically acceptable auxiliary materials.
Preferably, the dosage form of the anti-breast cancer medicament is injection.
The invention provides a new application of Fe-N-C monatomic nanoenzyme, namely the application of Fe-N-C monatomic nanoenzyme in preparing antitumor drugs. The experimental result shows that the Fe-N-C monatomic nanoenzyme can catalyze H2O2Formation of OH-Under the irradiation of 808nm ultraviolet laser, the light energy can be converted into heat energy, and the tumor cells are killed and killed by the dual functions. The invention provides a new treatment scheme for tumor treatment and has wide application prospect.
Drawings
FIG. 1 is a schematic structural diagram of a Fe-N-C monatin nanoenzyme of the present invention;
FIG. 2 is a diagram showing the photothermal conversion effect of Fe-N-C monatomic nanoenzyme aqueous solutions of different concentrations according to the present invention;
FIG. 3 is a graph showing the effect of different concentrations of Fe-N-C monatin nanoenzymes in example 3 of the present invention in simulating peroxidase;
FIG. 4 shows the inhibition of the activity of triple negative breast cancer cells by Fe-N-C monatin nanoenzyme in combination with laser irradiation in example 4 of the present invention;
FIG. 5 shows that Fe-N-C monatin nanoenzymes with different concentrations in example 5 of the present invention catalyze OH in tumor cells in vitro-The formation diagram of (2);
FIG. 6 shows the tumor apoptosis of triple negative breast cancer MDA-231 treated with Fe-N-C monatomic nanoenzymes of different concentrations in example 6 of the present invention.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments of the present invention, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the 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.
The invention provides application of Fe-N-C monatomic nanoenzyme in preparation of antitumor drugs.
In the invention, the Fe-N-C monatomic nanoenzyme has 1 iron atom, 4 nitrogen atoms and a plurality of carbon atoms. The Fe-N-C monatomic nanoenzyme has a structure shown in figure 1. FIG. 1 is a schematic structural diagram of the Fe-N-C monatomic nanoenzyme of the present invention.
The preparation method of the Fe-N-C monatin nanoenzyme is not particularly limited, and in some embodiments of the invention, the Fe-N-C monatin nanoenzyme can be prepared according to the method described in ACS Catal.2020,10, 6422-6429.
The invention uses Fe-N-C single atom nano enzyme, TMB and H2O2The first mixed solution is prepared, the activity of Fe-N-C simulated peroxidase is researched, and the result shows that Fe-N-C monatomic nanoenzyme with different concentrations can catalyze H2O2Formation of OH-An absorption peak at 650nm was observed by TMB. In certain embodiments of the invention, the concentration of the Fe-N-C monatin nanoenzyme in the first mixed solution is 0.5. mu.g/mL to 10 mg/mL.
The invention prepares MDA-231 cells, Fe-N-C monatin nanoenzyme and TMB into a second mixed solution, and researches OH of Fe-N-C monatin nanoenzyme in-vitro catalysis of tumor cells-The formation of (c). The result shows that in the triple negative breast cancer cell MDA-231, Fe-N-C monoatomic nanoenzyme with different concentrations can catalyze and generate OH-An absorption peak at 650nm was observed by TMB. In some embodiments of the invention, the second mixed solution is Fe-N-C monoThe concentration of the atomic nano enzyme is 0-100 mug/mL.
Therefore, the invention provides the application of Fe-N-C monatin nanoenzyme as peroxidase and the application of Fe-N-C monatin nanoenzyme in preparing medicines for reducing H in tumor cells2O2Application in medicine.
The invention dissolves Fe-N-C monatomic nanoenzyme in water, promotes the dissolution by using a vibration and ultrasonic method to obtain 20 mu g/mL-1 mg/mL Fe-N-C monatomic nanoenzyme aqueous solution, irradiates by using 808nm laser, and records the change of temperature per minute. Researches show that after the Fe-N-C monatomic nanoenzyme aqueous solution is irradiated by laser of 808nm, the Fe-N-C monatomic nanoenzyme aqueous solution with different concentrations can convert light energy into heat energy, and the temperature is increased. Therefore, the invention provides the application of the Fe-N-C monoatomic nanoenzyme as a photothermal conversion agent.
The three-negative breast cancer cell MDA-231 is inoculated into a 96-well plate containing a culture solution (10% FBS RPMI-1640, Gibco), about 1 ten thousand per well is added, after cells adhere to the wall within 12-24 h, Fe-N-C monoatomic nanoenzyme with different concentrations is respectively added into cell supernatant, laser irradiation is given, then the cell supernatant of each well is sucked out, 90 mu L of the culture solution and 10 mu L of LCCK-8 are added, and after 12h of action, an absorption value under the wavelength of 450nm is detected by a spectrophotometer. The result shows that the Fe-N-C monatomic nanoenzyme can obviously inhibit the survival of the triple-negative breast cancer cells through the combined laser irradiation in the triple-negative breast cancer cells MDA-231. In certain embodiments of the invention, the Fe-N-C monatin nanoenzyme is used in an amount of 3.125. mu.g/mL to 100 mg/mL. In some embodiments of the present invention, the laser irradiation uses a laser wavelength of 808nm, and the irradiation power of the laser is 1.0W cm-2The irradiation time was 10 min.
Therefore, the invention also provides the application of the Fe-N-C monatomic nanoenzyme in the preparation of anti-breast cancer drugs. Specifically, the method comprises the following steps:
free radicals generated by Fe-N-C monatomic nanoenzyme through photothermal action and Fenton reaction kill breast cancer cells.
In certain embodiments of the invention, the breast cancer cell is triple negative breast cancer cell MDA-231.
In certain embodiments of the invention, the anti-breast cancer drug comprises a Fe-N-C monatin nanoenzyme and a pharmaceutically acceptable excipient.
In certain embodiments of the present invention, the dosage form of the anti-breast cancer drug is an injection. In some embodiments, the injection can be an injection or a powder injection.
The invention provides a new application of Fe-N-C monatomic nanoenzyme, namely the application of Fe-N-C monatomic nanoenzyme in preparing antitumor drugs. The experimental result shows that the Fe-N-C monatomic nanoenzyme can catalyze H2O2Formation of OH-Under the irradiation of 808nm ultraviolet laser, the light energy can be converted into heat energy, and the tumor cells are killed and killed by the dual functions. The invention provides a new treatment scheme for tumor treatment and has wide application prospect.
The source of the above-mentioned raw materials is not particularly limited in the present invention, and may be generally commercially available.
In order to further illustrate the present invention, the following examples are given to describe the application of a Fe-N-C monatin nanoenzyme of the present invention in detail, but should not be construed as limiting the scope of the present invention.
Example 1
Preparing Fe-N-C monatomic nanoenzyme:
2.5mmol of ferric nitrate was dissolved in 30mL of a methanol solution containing 10mmol of 2-methylimidazole, stirred for 30min, and 500mg of KCl was added to the resulting solution, followed by drying at 80 ℃ for 6 h. Heating the obtained brown product to 750 deg.C at a speed of 5 deg.C/min under argon, calcining for 2 hr to obtain black powder, and adding 0.1mol/LH into the black powder2SO4The solution and ultrapure water were washed to remove the Fe nanoparticles and KCl template. And drying the obtained product in a vacuum oven at 85 ℃ to obtain the Fe-N-C monoatomic nanoenzyme.
Example 2
Photothermal conversion of Fe-N-C monatomic nanoenzyme:
dissolving Fe-N-C monatomic nanoenzyme of example 1 in water, using a shaking and ultrasonic method to promote the dissolution to obtain Fe-N-C monatomic nanoenzyme aqueous solution, taking 1mL of Fe-N-C monatomic nanoenzyme aqueous solution with different concentrations (20 mug/mL, 50 mug/mL, 100 mug/mL, 500 mug/mL and 1mg/mL respectively), irradiating with 808nm laser, recording the change of temperature every minute, and continuously recording for 12 min. The results are shown in FIG. 2. FIG. 2 is a diagram showing the photothermal conversion effect of Fe-N-C monatomic nanoenzyme aqueous solutions of different concentrations according to the present invention. As can be seen from FIG. 2, after the Fe-N-C monatomic nanoenzyme aqueous solution with different concentrations is irradiated by laser with 808nm, the optical energy can be converted into heat energy, and the temperature is increased.
Example 3
Fe-N-C monatomic nanoenzyme simulated peroxidase catalyzed H2O2Formation of OH-：
Preparing 2mL of test solution (HAc-NaAc 0.1mol/L, pH 3.0) containing Fe-N-C monatomic nanoenzyme, TMB and H2O2The final concentrations of Fe-N-C monatin nanoenzymes were 0.5. mu.g/mL, 1. mu.g/mL, 2. mu.g/mL, 5. mu.g/mL, 10. mu.g/mL, respectively, the final concentration of TMB was 0.192mg/mL, H2O2The final concentration of (3) was 3 mmol/L. Standing for 2min after mixing, and detecting the absorption wavelength with an ultraviolet spectrophotometer, the result is shown in FIG. 3. FIG. 3 is a graph showing the effect of different concentrations of Fe-N-C monatin nanoenzymes in example 3 of the present invention in simulating peroxidase. As can be seen from FIG. 3, Fe-N-C monatin nanoenzymes with different concentrations can catalyze H2O2Formation of OH-An absorption peak at 650nm was observed by TMB.
Example 4
The Fe-N-C monatomic nanoenzyme has the in vitro anti-tumor effect:
inoculating triple negative breast cancer cell MDA-231 (from China university of sciences academy of Life sciences) into a 96-well plate containing a culture solution (10% FBS RPMI-1640, Gibco), adding Fe-N-C monatin nanoenzymes with different concentrations into cell supernatant respectively after cells are attached for 12-24 h, wherein the concentrations of the Fe-N-C monatin nanoenzymes in the cell supernatant are respectively 0 mu g/mL, 3.125 mu g/mL, 6.25 mu g/mL, 12.5 mu g/mL, 25 mu g/mL, 50 mu g/mL and 100 mu g/mL (each group is provided with 3 multiple wells), acting for 4h, and giving 808nm laser 1.0W cm-2Irradiating for 10min with power. Aspirating the cell supernatant from each well, addingAfter the reaction in 90. mu.L of the culture medium (10% FBS RPMI-1640, Gibco) and 10. mu.L of LCCK-8 for 12 hours, the absorbance at a wavelength of 450nm was measured with a spectrophotometer, and the results are shown in FIG. 4. FIG. 4 shows the inhibition of the activity of triple negative breast cancer cells by Fe-N-C monatin nanoenzyme in combination with laser irradiation in example 4 of the present invention; wherein, Water: water; laser: 808nm laser. As can be seen from FIG. 4, in the triple negative breast cancer cell MDA-231, the Fe-N-C monatomic nanoenzyme combined with laser irradiation can significantly inhibit the survival of the triple negative breast cancer cell MDA-231.
Example 5
Fe-N-C monatomic nanoenzyme for catalyzing OH in tumor cells in vitro-Forming:
2mL of a test solution (HAc-NaAc 0.1mol/L, pH 3.0) containing about 20X 10 MDA-231 cells was prepared4Fe-N-C single atom nano enzyme and TMB. The final concentrations of the Fe-N-C monatomic nanoenzyme are 0 mug/mL, 10 mug/mL, 25 mug/mL, 50 mug/mL and 100 mug/mL respectively, and the final concentration of the TMB is 0.192 mg/mL. After mixing, the mixture was left to stand for 5min, and the absorption wavelength was measured with an ultraviolet spectrophotometer, and the result is shown in FIG. 5. FIG. 5 shows that Fe-N-C monatin nanoenzymes with different concentrations in example 5 of the present invention catalyze OH in tumor cells in vitro-To form a figure. As can be seen from FIG. 5, in the triple negative breast cancer MDA-231, Fe-N-C monatin nanoenzyme with different concentrations can catalyze the generation of OH-An absorption peak at 650nm was observed by TMB.
Example 6
After the Fe-N-C monatomic nanoenzyme is irradiated by combining laser with 808nm, the apoptosis of tumor cells is increased in vitro:
collecting triple negative breast cancer cell MDA-23120 × 104One was placed in a 1.5mL EP tube, and 4 sets were set: control (Control) group, Fe-N-C group, Laser group, Fe-N-C + Laser group. The control group is blank control group (10% FBS RPMI-1640, Gibco), Fe-N-C group is added with Fe-N-C monatomic nanoenzyme, wherein the concentration of Fe-N-C monatomic nanoenzyme is 50mg/mL, and the action is 4 h; laser irradiation of the Laser group is carried out with 808nm Laser of 1.0W cm-2Irradiating for 10min with power; Fe-N-C + Laser group is added with Fe-N-C monatomic nanoenzyme, wherein the concentration of the Fe-N-C monatomic nanoenzyme is 50mg/mL, the Fe-N-C monatomic nanoenzyme acts for 4 hours, and the Laser with the wavelength of 808nm is 1.0Wcm-2Irradiating for 10min with power. The cells after the 4 groups of treatment are centrifuged at 4000rpm for 4min, 500 muL of 1 XBuffer in the apoptosis detection kit is taken for resuspending the cells, 5 muL of Lannexin V and 10 muL of 7-AAD are added, and the cells are protected from light for 5 min. And (3) detecting by an up-flow machine (FACSVerse flow cytometer, BD). The results are shown in FIG. 6. FIG. 6 shows the tumor cell apoptosis after different treatments are performed on triple negative breast cancer MDA-231 in example 6 of the present invention, and it can be seen from FIG. 6 that in triple negative breast cancer MDA-231, Fe-N-C monatomic nanoenzyme 50 μ g/mL induces apoptosis, and increases 808nm laser irradiation, which significantly enhances apoptosis.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (10)
- The application of Fe-N-C monatomic nanoenzyme in preparing antitumor drugs.
- The application of Fe-N-C monatomic nanoenzyme as peroxidase.
- Preparation of Fe-N-C monatin nanoenzyme for reducing H in tumor cells2O2Application in medicine.
- And 4, application of Fe-N-C monatomic nanoenzyme as a photothermal conversion agent.
- 5, application of Fe-N-C monatomic nanoenzyme in preparation of anti-breast cancer drugs.
- 6. The use according to claim 5, comprising: free radicals generated by Fe-N-C monatomic nanoenzyme through photothermal action and Fenton reaction kill breast cancer cells.
- 7. The use according to claim 6, wherein the photothermal action is at a laser wavelength of 808 nm;the irradiation power of the laser is 1.0W cm-2The irradiation time was 10 min.
- 8. The use of claim 6, wherein the amount of the Fe-N-C monatin nanoenzyme is 3.125 μ g/mL to 100 mg/mL.
- 9. The use of claim 5, wherein the anti-breast cancer drug comprises Fe-N-C monatin nanoenzyme and a pharmaceutically acceptable excipient.
- 10. The use of claim 5, wherein the anti-breast cancer drug is in the form of an injection.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202110296350.7A CN113181358A (en) | 2021-03-19 | 2021-03-19 | Application of Fe-N-C monatomic nanoenzyme |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115463151A (en) * | 2022-08-18 | 2022-12-13 | 湖北文理学院 | Nano enzyme, preparation method and application thereof, and bacteriostatic agent |
| CN115709099A (en) * | 2022-10-28 | 2023-02-24 | 湖南大学 | Monoatomic nanoenzyme Fe-N-C loaded polyvinylidene fluoride composite membrane and preparation method and application thereof |
-
2021
- 2021-03-19 CN CN202110296350.7A patent/CN113181358A/en active Pending
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| Title |
|---|
| FEI HE ET AL: ""Fe−N−C Artificial Enzyme: Activation of Oxygen for Dehydrogenation and Monoxygenation of Organic Substrates under Mild Condition and Cancer Therapeutic Application"", 《ACS APPL. MATER. INTERFACES》 * |
| LEI JIAO ET AL: ""Densely Isolated FeN4 Sites for Peroxidase Mimicking"" * |
| LEI JIAO ET AL: ""Densely Isolated FeN4 Sites for Peroxidase Mimicking"", 《ACS CATAL》 * |
| MINFENG HUO ET AL: ""Nanocatalytic Tumor Therapy by Single-Atom Catalysts"" * |
| MINFENG HUO ET AL: ""Nanocatalytic Tumor Therapy by Single-Atom Catalysts"", 《ACS NANO》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115463151A (en) * | 2022-08-18 | 2022-12-13 | 湖北文理学院 | Nano enzyme, preparation method and application thereof, and bacteriostatic agent |
| CN115463151B (en) * | 2022-08-18 | 2023-09-19 | 湖北文理学院 | Nano-enzyme, preparation method and application thereof, and bacteriostat |
| CN115709099A (en) * | 2022-10-28 | 2023-02-24 | 湖南大学 | Monoatomic nanoenzyme Fe-N-C loaded polyvinylidene fluoride composite membrane and preparation method and application thereof |
| CN115709099B (en) * | 2022-10-28 | 2024-01-26 | 湖南大学 | Polyvinylidene fluoride composite film loaded with monoatomic nano enzyme Fe-N-C as well as preparation method and application thereof |
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