A single-iron atom nanoscale enzyme preparation loaded with sodium hydrosulfide and a preparation method and application thereof

Abstract

The application provides an iron monatomic nanoscale enzyme preparation loaded with sodium hydrosulfide and a preparation method and application thereof, to construct an iron monatomic nanoscale enzyme preparation loaded with sodium hydrosulfide by taking the iron monatomic nanoscale enzyme as a drug carrier loaded with sodium hydrosulfide and taking hyaluronic acid as an encapsulating agent and a targeting agent. On one hand, hydrogen sulfide gas released by sodium hydrosulfide can inhibit catalase activity in tumor cells, so as to enhance the chemical kinetic treatment effect of the iron monatomic nanoscale enzyme; on the other hand, an increase in active oxygen can cause mitochondrial dysfunction, so as to reduce ATP synthesis and further reduce the expression of heat shock proteins, and promote the mild photothermal therapy based on the iron monatomic nanoscale enzyme.

Description

TECHNICAL FIELD

[0001] The application relates to the technical field of nanomaterials, in particular to a sodium hydrosulfide-loaded iron monatomic nanoscale enzyme preparation and a preparation method and application thereof. BACKGROUND

[0002] Monatomic nanoscale enzymes (SAzymes) have the maximum atomic utilization rate, clear metal center and coordination environment, and have become a nanomedical frontier for tumor-specific treatment. There are two induction pathways for SAzymes-based cancer treatment: one is the classic chemodynamic therapy (CDT) mode, that is, through the Fenton reaction, the overexpressed H2O2 in the tumor is converted into highly toxic hydroxyl radicals (·OH), which can destroy DNA, proteins and lipids and other biological molecules, and finally lead to cell death. In addition, due to the photo-thermal conversion characteristics of the single-atom-anchored amorphous carbon carrier, the photothermal therapy (PTT) based on SAzymes becomes another way to eradicate tumors. However, its treatment effect is usually interfered by the lack of H2O2 and the up-regulation of heat shock proteins (HSPs). At present, the method for simultaneously enhancing the CDT and PTT efficiency of SAzymes to maximize the tumor treatment effect is rare.

[0003] High-temperature PTT (>50℃) is a way of converting light energy into heat energy for tumor lesion ablation, which has high spatiotemporal accuracy and less invasiveness. However, the heat diffusion generated by high-temperature PTT inevitably causes inflammation and damage to adjacent healthy tissues. In order to solve these problems, mild PTT (<45℃) as an alternative method is more promising for safe treatment of cancer, thereby avoiding damage to normal cells. Unfortunately, cell heat tolerance is often caused by overexpression of HSPs. Current research has found that H2S gas can down-regulate the activity of mitochondrial respiratory cytochrome c oxidase, thereby blocking the biosynthesis of adenosine triphosphate (ATP) energy to reduce HSPs expression, and finally improve the efficacy of mild PTT. There are mainly two strategies for down-regulating HSPs to increase the sensitivity of tumor cells to mild PTT: (1) inactivating or reducing the existing HSPs; (2) reducing ATP energy supply to inhibit HSPs expression. Compared with the first strategy, ATP depletion cuts off the energy supply for protein biosynthesis, which will be a more attractive solution to bypass heat shock protein-mediated heat tolerance. Therefore, it is a challenging work to develop a new way to efficiently reduce ATP production and fundamentally inhibit the expression of HSPs. SUMMARY

[0004] To solve the above technical problems and maximize the synergistic therapeutic efficiency based on SAzymes, the application provides a sodium hydrosulfide-loaded iron monatomic nanoscale enzyme preparation, which is prepared by coating the surface of iron monatomic nanoscale enzymes with hyaluronic acid as an encapsulating agent and a targeting agent, and assembling the iron monatomic nanoscale enzymes into the nanoscale preparation.

[0005] The application also provides a preparation method of the sodium hydrosulfide-loaded iron monatomic nanoscale enzyme preparation, which comprises the following steps:

[0006] Step one: preparation of porous iron monatomic nanoscale enzymes: dissolve ferric nitrate and zinc nitrate in a solvent to form solution 1, dissolve 2-methylimidazole in a solvent to form solution 2, mix, stir, age, purify and dry solution 1 and solution 2 to obtain Fe / Zn-ZIFs precursor powder, transfer the Fe / Zn-ZIFs precursor powder to a porcelain boat, and obtain the porous iron monatomic nanoscale enzymes by high-temperature pyrolysis in a tube furnace.

[0007] Step two: preparation of sodium hydrosulfide-loaded iron monatomic nanoscale enzymes: prepare an aqueous solution containing sodium hydrosulfide, disperse the porous iron monatomic nanoscale enzymes prepared in step one into the aqueous solution containing sodium hydrosulfide, incubate overnight, and obtain the sodium hydrosulfide-loaded iron monatomic nanoscale enzymes by centrifugal treatment after the incubation is completed.

[0008] Step three: preparation of sodium hydrosulfide-loaded iron monatomic nanoscale enzyme preparation: disperse hyaluronic acid in water to obtain a solution containing hyaluronic acid, disperse the sodium hydrosulfide-loaded iron monatomic nanoscale enzymes obtained in step two in the aqueous solution containing hyaluronic acid and stir, and obtain the sodium hydrosulfide-loaded iron monatomic nanoscale enzyme preparation by centrifugation after the stirring is completed.

[0009] The application also provides an application of the sodium hydrosulfide-loaded iron monatomic nanoscale enzyme preparation in the preparation of an antitumor drug.

[0010] Beneficial effects

[0011] Monatomic nanoszymes are considered as a promising nanodrug for tumor therapy due to their excellent peroxidase activity, glutathione oxidase (GSHOx) activity, photothermal conversion performance and biocompatibility. The prepared Fe / SAzymes can convert the overexpressed H2O2 in tumor into highly toxic ·OH through Fenton reaction in the micro-acidic and H2O2-rich environment of tumor, thereby destroying the intracellular biomolecules such as DNA, proteins and lipids, and ultimately leading to cancer cell death. However, in reality, the intracellular H2O2 is still insufficient despite its overexpression. The Fe / SAzymes are loaded with sodium hydrosulfide and coated with hyaluronic acid to obtain a nanoreagent. After being internalized by cells, the released H2S can inhibit the catalase activity in cancer cells, thereby reducing the consumption of endogenous H2O2 and increasing the amount of ·OH generated by monatomic nanoszymes to enhance the CDT efficacy. The experimental results show that the Fe / SAzymes@NaHS@HA treatment group exhibits stronger antitumor activity than the Fe / SAzymes@HA treatment group alone, which confirms that H2S can promote the CDT efficacy of Fe / SAzymes.

[0012] The synthesis of heat shock proteins is directly positively correlated with the ATP content, and ATP depletion cuts off the energy supply for protein biosynthesis, which will be a more attractive solution to avoid heat resistance mediated by heat shock proteins. The prepared Fe / SAzymes have another important feature of excellent photothermal conversion performance due to the amorphous carbon structure of the carrier. The temperature change curve of different Fe / SAzymes concentrations under 1064 nm laser irradiation can prove that it is an excellent photothermal conversion nanomaterial, and the calculated photothermal conversion efficiency is 27.59%, which provides a photothermal ablation approach for cancer treatment. The excess ·OH generated by the above H2S-assisted Fe / SAzymes can not only enhance the CDT efficacy of Fe / SAzymes, but also cause mitochondrial dysfunction, leading to a decrease in ATP production, and ultimately hinder the synthesis of heat shock proteins, thereby enhancing the mild PTT of Fe / SAzymes. The mild PTT cooperates with CDT to achieve significant inhibition of tumor cell growth. In summary, compared with the use of Fe / SAzymes alone, the combined use of Fe / SAzymes and NaHS significantly enhances the tumor treatment effect, indicating that H2S can not only promote the CDT efficacy of Fe / SAzymes, but also promote the mild PTT of Fe / SAzymes, and ultimately effectively improve the synergistic cancer treatment efficacy of Fe / SAzymes.

[0013] The embodiment of the application takes porous iron single atom nanoszyme as a carrier, sodium hydrosulfide enters the pores of the porous iron single atom nanoszyme after being co-incubated with the porous iron single atom nanoszyme, and hyaluronic acid is used as an encapsulating agent and a targeting agent to coat the surface of the iron single atom nanoszyme to assemble into a nano preparation. The H2S released by NaHS enhances the tumor treatment effect of Fe / SAzymes. On the one hand, H2S enhances the CDT effect of Fe / SAzymes by inhibiting the activity of catalase, and on the other hand, the generated ROS causes mitochondrial dysfunction to reduce ATP synthesis and thus reduce the content of HSPs. Finally, under the irradiation of 1064 nm laser, the mild PTT effect of Fe / SAzymes is enhanced. BRIEF DESCRIPTION OF DRAWINGS

[0014] Figure 1 The transmission electron microscopy characterization of the morphology structure of Fe / SAzymes in an embodiment of the application is shown;

[0015] Figure 2 The high-angle annular dark field scanning transmission electron microscopy characterization of Fe / SAzymes in an embodiment of the application is shown, and the results of element superposition analysis of iron (Fe), carbon (C), nitrogen (N) and the three elements are shown;

[0016] Figure 3 The high-angle annular dark field scanning transmission electron microscopy characterization of single atom Fe in an embodiment of the application is shown

[0017] Figure 4 The X-ray photoelectron spectroscopy detection result of Fe / SAzymes in an embodiment of the application is shown;

[0018] Figure 5 The nitrogen isothermal adsorption-desorption curve of the mesoporous material Fe / SAzymes in an embodiment of the application is shown;

[0019] Figure 6 The peroxidase activity test result of Fe / SAzymes in an embodiment of the application is shown;

[0020] Figure 7 The photothermal conversion performance test result of Fe / SAzymes in an embodiment of the application is shown;

[0021] Figure 8 The Zeta potential of Fe / SAzymes, Fe / SAzymes@NaHS and Fe / SAzymes@NaHS@HA in an embodiment of the application is shown;

[0022] Figure 9 The H2S release test result of Fe / SAzymes@NaHS@HA in an embodiment of the application is shown;

[0023] Figure 10Fig. 1 shows the cell viability of PANC-1 cells treated with different drugs according to an embodiment of the present application. DETAILED DESCRIPTION

[0024] The preferred embodiments of the present application will be described in detail hereinafter with reference to the accompanying drawings. It is to be understood that the following examples are given for illustrative purposes only and are not intended to limit the scope of the present application. Those skilled in the art can make various modifications and substitutions to the present application without departing from the spirit and scope of the present application.

[0025] The experimental methods used in the following examples are conventional methods unless otherwise specified.

[0026] The materials, reagents, etc. used in the following examples can be obtained commercially unless otherwise specified.

[0027] An embodiment of the present application provides a sodium hydrosulfide-loaded iron monatomic nanoscale enzyme preparation, which is a nanoscale preparation assembled by coating hyaluronic acid on the surface of iron monatomic nanoscale enzymes as an encapsulating agent and targeting agent, and the sodium hydrosulfide enters the pores of the porous iron monatomic nanoscale enzyme after being co-incubated with the porous iron monatomic nanoscale enzyme. The transmission electron microscope characterization of the morphology structure of the sodium hydrosulfide-loaded iron monatomic nanoscale enzyme preparation Fe / SAzymes is shown in Fig. 1. Figure 1

[0028] An embodiment of the present application provides a preparation method of the sodium hydrosulfide-loaded iron monatomic nanoscale enzyme preparation, which comprises the following steps:

[0029] Step 1: Preparation of porous iron monatomic nanoscale enzyme: dissolve ferric nitrate and zinc nitrate in a solvent to form solution 1, and dissolve 2-methylimidazole in a solvent to form solution 2. After mixing, stirring, aging, purification and drying of solution 1 and solution 2, Fe / Zn-ZIFs precursor powder is obtained. The Fe / Zn-ZIFs precursor powder is transferred to a porcelain boat and subjected to high-temperature pyrolysis in a tube furnace to obtain porous iron monatomic nanoscale enzyme.

[0030] Step 2: Preparation of sodium hydrosulfide-loaded iron monatomic nanoscale enzyme: prepare an aqueous solution containing sodium hydrosulfide, and disperse the porous iron monatomic nanoscale enzyme Fe / SAzymes prepared in step 1 into the aqueous solution containing sodium hydrosulfide. Incubate overnight, and then centrifuge to obtain sodium hydrosulfide-loaded iron monatomic nanoscale enzyme.

[0031] ​Step three: preparation of sodium hydrosulfide loaded iron monatomic nanoscale enzyme preparation: hyaluronic acid was dispersed in water to obtain a solution containing hyaluronic acid, and the sodium hydrosulfide loaded iron monatomic nanoscale enzyme obtained in step two was dispersed in the hyaluronic acid-containing aqueous solution and stirred, and after the stirring was completed, the sodium hydrosulfide loaded iron monatomic nanoscale enzyme preparation was obtained by centrifugation.

[0032] In an embodiment, in step one, the final concentration of ferric nitrate is 0.30-0.50 mg / ml, the final concentration of zinc nitrate is 4.00-7.00 mg / ml, and the final concentration of 2-methylimidazole is 12.00-15.00 mg / ml; the solvent is methanol; the stirring time is 1 h, the aging time is 24 h, and the drying temperature is 60°C; the high-temperature pyrolysis conditions are as follows: under a nitrogen atmosphere, the temperature is raised to 800°C at a rate of 5°C / min, and the target temperature is maintained for 2 h.

[0033] In an embodiment, in step two, the concentration of sodium hydrosulfide is 16 mg / ml, and the concentration of iron monatomic nanoscale enzyme is 1 mg / ml; the incubation conditions are 25°C; and the centrifugation conditions are 16000 rpm for 8 min.

[0034] In an embodiment, in step three, the concentration of hyaluronic acid is 10 mg / ml; the stirring conditions are 1 h at room temperature, and the centrifugation conditions are 16000 rpm for 8 min.

[0035] An embodiment of the present application provides the use of the sodium hydrosulfide loaded iron monatomic nanoscale enzyme preparation in the preparation of an antitumor drug.

[0036] In an embodiment, the concentration of the nanoscale preparation ranges from 0 μg / mL to 50 μg / mL.

[0037] In an embodiment, a 1064 nm near-infrared laser is used for irradiation treatment of the nanoscale preparation.

[0038] Preparation of nanocarriers:

[0039] 1. Preparation of porous iron monatomic nanoscale enzyme (Fe / SAzymes)

[0040] First, 0.08g of Fe(NO₃)₃·9H₂O and 1.116g of Zn(NO₃)₂·6H₂O were dissolved in 100mL of methanol to form solution 1. 2.608g of 2-methylimidazole was dissolved in 100mL of methanol to form solution 2. Solution 1 was poured into solution 2 at room temperature and stirred vigorously for 1h. The mixture was then aged for 24h. Finally, the yellow precipitate was purified five times with methanol and dried at 60°C to obtain the precursor Fe-Zn ZIFs. Finally, the synthesized Fe-Zn ZIFs precursor was placed in a porcelain boat and heated at 5°C / min to 800°C under a nitrogen atmosphere. The temperature was then maintained at this temperature for 2h to obtain Fe / SAzymes.

[0041] 2. Preparation of iron single-atom nanozymes loaded with sodium hydrosulfide (Fe / SAzymes@NaHS)

[0042] Weigh 1 mg of the Fe / SAzymes obtained in step 1 into 1 mL of PBS (pH 7.4) and mix thoroughly by sonication. Next, weigh 16 mg of NaHS into the solution and incubate overnight. After incubation, centrifuge (16,000 rpm, 8 min) to obtain the Fe / SAzymes@NaHS.

[0043] 3. Preparation of iron single-atom nanozyme preparation loaded with sodium hydrosulfide (Fe / SAzymes@NaHS@HA)

[0044] Weigh 5 mg of hyaluronic acid (HA) into 500 μL of the 1 mg / mL Fe / SAzymes@NaHS solution (step 2). Stir slowly with a magnetic stirrer for 1 hour to coat the nanozyme with HA. After stirring, centrifuge to obtain the sodium hydrosulfide-loaded iron single-atom nanozyme preparation.

[0045] Characterization of Nanocarriers:

[0046] 1. Synthesis and characterization of Fe / SAzymes

[0047] like Figure 1 As shown in Figure 2, transmission electron microscopy (TEM) images show that FeSAzymes have good geometric morphology and an average diameter of about 120 nm. In order to confirm whether Fe exists in the Fe / SAzymes structure, elemental analysis was performed. Figure 2 The results showed that Fe, C and N elements existed in FeSAzymes. Figure 3As shown, high-angle annular dark-field scanning transmission electron microscopy (HAADF-STEM) images confirmed the presence of monodisperse iron atoms, thus the successful synthesis of Fe / SAzymes. Second, through X-ray photoelectron spectroscopy (XPS), the N 1s spectrum of Fe / SAzymes showed three peaks located at 398.4 eV (pyridine N), 400.5 eV (pyrrole N), and 402.2 eV (graphitic N) Figure 4 ) Among them, pyridine N helps to stabilize monodisperse Fe atoms, thus facilitating the catalytic activity of Fe / SAzymes. As shown in Figure 5 , N2 adsorption / desorption experiments were performed on Fe / SAzymes, Figure 5 , which showed an average pore size of 1.39 nm, confirming the presence of a porous structure.

[0048] 2. Peroxidase (POD) activity and photothermal conversion performance test of Fe / SAzymes

[0049] 3, 3', 5, 5'-tetramethylbenzidine (TMB) and ·OH typical colorimetric reaction verified the POD activity of Fe / SAzymes, which can catalyze TMB to generate blue oxidized TMB (oxTMB) and produce obvious ultraviolet-visible characteristic absorption peak at 652 nm. Different combinations of TMB analysis were explored, as shown in Figure 6 A, Fe / SAzymes had obvious enhancement of POD catalytic activity compared with CN. In addition, Figure 6 B shows that Fe / SAzymes has acid-dependent POD activity at different pH values (5.0, 6.0 and 7.4), which indicates that Fe / SAzymes tends to convert H2O2 to abundant ·OH in a slightly acidic environment. The temperature rise curve of Fe / SAzymes solution at different concentrations proves that Fe / SAzymes can be used as an excellent photothermal conversion nanomaterial ( Figure 7 ), and the photothermal conversion efficiency is calculated to be 27.59%.

[0050] 3. Characterization test of Fe / SAzymes@NaHS@HA and H2S release performance test.

[0051] As shown in Figure 8As shown in the figure, the Zeta potential of Fe / SAzymes is -11.43mV. After NaHS enters the interior of Fe / SAzymes, the Zeta potential of Fe / SAzymes@NaHS is -15.80mV, which indicates that the loading of NaHS is successful. In order to prevent the leakage of NaHS, a layer of hyaluronic acid (HA) is wrapped on the surface of Fe / SAzymes, and the Zeta potential changes from the original -15.80mV to -17.52mV, which indicates that the assembly of Fe / SAzymes@NaHS@HA nanoformulation is successful. Next, the H2S detection kit is used, that is, H2S reacts with N,N-dimethyl-p-phenylenediamine and ammonium ferric sulfate to generate methylene blue. Methylene blue has a maximum absorption peak around 665nm, as shown in the figure. Figure 9 As shown in the figure, Fe / SAzymes@NaHS@HA has obvious characteristic absorption peaks after being treated with the kit, proving that it can release H2S gas.

[0052] Application of nanocarriers:

[0053] The cytotoxicity was detected using a standard cell counting kit-8 (CCK-8). PANC-1 cells were cultured in 96-well plates at a cell density of 1×10 4 / well (100μL RPMI-1640 medium), cultured for 12h until the cells adhered. Next, the cells were incubated with (1) PBS; (2) Fe / SAzymes@HA; (3) Fe / SAzymes@HA+laser; (4) Fe / SAzymes@NaHS@HA; (5) Fe / SAzymes@NaHS@HA+laser. After incubation of the drugs with the cells for 12h, the cells were illuminated with a 1064nm laser (0.58W cm -2 PANC-1 cells were irradiated for 10 min (15 min). An infrared camera monitored the maximum temperature during illumination to maintain it below 45°C. After 24 hours of incubation, the culture medium was removed and the cells were incubated with fresh culture medium containing 10 μL of CCK-8 for 1 hour. Cell viability was assessed by measuring the absorbance of CCK-8 at 450 nm.

[0054] like Figure 10 As shown in Figure A, the antitumor activity of Fe / SAzymes against PANC-1 cells increased with the increase of Fe / SAzymes concentration from 0 to 140 μg / mL, which was attributed to the slightly acidic pH and abundant endogenous H2O2 in the tumor microenvironment. Then, Fe / SAzymes@NaHS@HA was used to study the synergistic therapeutic effect between H2S gas and single-atom nanozymes. The results showed that Fe / SAzymes@NaHS@HA treatment exhibited stronger antitumor activity than Fe / SAzymes@HA treatment alone, which confirmed that H2S can promote the CDT efficacy of Fe / SAzymes (Figure 10 In addition, the cell viability after Fe / SAzymes@HA treatment was less than 65% under 1064 nm laser irradiation, while the cell viability after Fe / SAzymes@NaHS@HA treatment was reduced to less than 40%. Compared with Fe / SAzymes alone, the combination of Fe / SAzymes and NaHS significantly enhanced the mild PTT (<45℃) effect of Fe / SAzymes, which indicated that H2S not only promoted the CDT effect of Fe / SAzymes, but also promoted its mild PTT, and finally effectively improved the synergistic cancer treatment effect of Fe / SAzymes.

[0055] In the embodiments of the present application, porous iron single-atom nanoszymes are used as carriers, sodium hydrosulfide enters the internal structure of the porous iron single-atom nanoszymes after co-incubation, and hyaluronic acid is used as an encapsulating agent and a targeting agent to coat the surface of the iron single-atom nanoszymes to assemble into a nanodrug. The H2S released by NaHS enhances the tumor treatment effect of Fe / SAzymes. On the one hand, H2S enhances the CDT effect of Fe / SAzymes by inhibiting the activity of catalase, and on the other hand, the generated ROS causes mitochondrial dysfunction, which reduces ATP synthesis and in turn reduces the content of HSPs, and finally enhances the mild PTT effect of Fe / SAzymes under 1064 nm laser irradiation. The above is only a preferred embodiment of the present application, and it should be noted that for those skilled in the art, several improvements and refinements can be made without departing from the principles of the present application, and these improvements and refinements should also be considered within the scope of protection of the present application.

Claims

1. A sodium hydrosulfide loaded iron monatomic nanoszyme formulation, characterized in that, The sodium hydrosulfide-loaded single-atom nanoscale enzyme preparation uses porous single-atom nanoscale enzyme as a carrier, and sodium hydrosulfide enters the pores of the porous single-atom nanoscale enzyme after being co-incubated with the porous single-atom nanoscale enzyme, and hyaluronic acid is used as an encapsulating agent and a targeting agent to coat the surface of the single-atom nanoscale enzyme to assemble into a nanoscale preparation; the preparation method of the sodium hydrosulfide-loaded single-atom nanoscale enzyme preparation comprises the following steps: Step one: preparing porous single-atom nanoscale enzyme: dissolving ferric nitrate and zinc nitrate in a solvent to form solution 1, dissolving 2-methylimidazole in a solvent to form solution 2, mixing, stirring, aging, purifying and drying solution 1 and solution 2 to obtain Fe / Zn-ZIFs precursor powder, and then transferring the Fe / Zn-ZIFs precursor powder to a porcelain boat and pyrolyzing in a tube furnace at high temperature to obtain porous single-atom nanoscale enzyme; Step two: preparing sodium hydrosulfide-loaded single-atom nanoscale enzyme: preparing an aqueous solution containing sodium hydrosulfide, dispersing the porous single-atom nanoscale enzyme prepared in step one into the aqueous solution containing sodium hydrosulfide, incubating overnight, and then centrifuging to obtain sodium hydrosulfide-loaded single-atom nanoscale enzyme; Step three: preparing sodium hydrosulfide-loaded single-atom nanoscale enzyme preparation: dispersing hyaluronic acid in water to obtain a solution containing hyaluronic acid, dispersing the sodium hydrosulfide-loaded single-atom nanoscale enzyme obtained in step two in the aqueous solution containing hyaluronic acid and stirring, and then centrifuging to obtain sodium hydrosulfide-loaded single-atom nanoscale enzyme preparation.

2. The sodium hydrosulfide-loaded iron single-atom nanozyme preparation according to claim 1, characterized in that In step one, the final concentration of ferric nitrate is 0.30-0.50 mg / ml, the final concentration of zinc nitrate is 4.00-7.00 mg / ml, and the final concentration of 2-methylimidazole is 12.00-15.00 mg / ml; the solvent is methanol; the stirring time is 1 h, the aging time is 24 h, and the drying temperature is 60℃; the high-temperature pyrolysis conditions are as follows: under a nitrogen atmosphere, the temperature is raised to 800℃ at a rate of 5℃ / min, and the target temperature is maintained for 2 h.

3. The sodium hydrosulfide supported iron monatomic nanosensor formulation of claim 1, wherein, In step two, the concentration of the aqueous solution containing sodium hydrosulfide is 16 mg / ml, and the concentration of the single-atom nanoscale enzyme is 1 mg / ml; the incubation conditions are 25℃; and the centrifugation conditions are 16000 rpm for 8 min.

4. The sodium hydrosulfide supported iron monatomic nanosensor formulation of claim 1, wherein, In step three, the concentration of the solution containing hyaluronic acid is 10 mg / ml; the stirring conditions are 1 h at 25℃, and the centrifugation conditions are 16000 rpm for 8 min.

5. The use of the sodium hydrosulfide-loaded single-atom nanoscale enzyme preparation of claim 1 in the preparation of an antitumor drug.

6. Use according to claim 5, characterized in that The sodium hydrosulfide-loaded single-atom nanoscale enzyme preparation is irradiated using a 1064 nm near-infrared laser.

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