MXene nano material with high SOD (superoxide dismutase) enzyme activity as well as preparation method and application of MXene nano material

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to an MXene nano material with high SOD (superoxide dismutase) enzyme activity as well as a preparation method and application thereof. The MXene nano material with high SOD (superoxide dismutase) enzyme activity, disclosed by the invention, comprises Mo2C MXene and a metal atom anchored on the Mo2C MXene. The MXene nano material with high SOD enzyme activity has the beneficial effects that the MXene nano material with high SOD enzyme activity has relatively high SOD enzyme activity and a strong ROS removal characteristic, and oxidative stress injury can be relieved; the compound has the effect of treating ROS related diseases such as traumatic brain injury (TBI), Parkinson's syndrome (PD) and chronic obstructive pulmonary disease.

Description

Technical Field

[0001] The present application belongs to the field of biomedicine technology, and specifically relates to a MXene nanomaterial with high SOD enzyme activity, a preparation method, and an application thereof. Background Art

[0002] Reactive oxygen species (ROS) mainly include superoxide radicals (O2 ·- ), hydroxyl radical (·OH), singlet oxygen ( 1 Oxygen-containing active species such as ROS (O2) and H2O2 have important physiological functions in living systems. Overexpression of ROS can lead to oxidative stress, causing local tissue damage and leading to the occurrence of various diseases. Although many antioxidant molecules and natural enzymes can reduce ROS levels, they often have limitations such as low ROS scavenging ability and insufficient stability in physiological environments. In recent years, the rise of enzyme-like catalytic or antioxidant nanomaterials with ROS modulation capabilities has provided a new way to alleviate oxidative stress damage. However, the superoxide dismutase (SOD) activity of current enzyme-like materials is not high. Many natural antioxidants, including astaxanthin, anthocyanins, and polyphenolic compounds, have been widely studied for their role in diseases caused by oxidative stress. However, these antioxidants often exhibit disadvantages such as limited efficiency in scavenging ROS, rapid metabolic clearance, and short-term activity, which limits their subsequent applications. Therefore, there is an urgent need to develop nanomaterials with high antioxidant enzyme activity and excellent biocompatibility. Summary of the Invention

[0003] The present application provides a MXene nanomaterial with high SOD enzyme activity, a preparation method and application thereof, aiming to solve the problems of low activity and poor biocompatibility of existing superoxide dismutase-like materials.

[0004] In a first aspect, the present application provides a MXene nanomaterial with high SOD enzyme activity, comprising Mo2C MXene and metal atoms anchored on the Mo2C MXene.

[0005] According to some embodiments of the MXene nanomaterial with high SOD enzyme activity described in the present application, the anchoring amount of metal atoms on the Mo2CMXene is 2 wt%-3 wt%.

[0006] According to some embodiments of the MXene nanomaterial with high SOD enzyme activity described in the present application, the metal atoms include one or more of copper atoms, iron atoms, platinum atoms, manganese atoms and zinc atoms.

[0007] According to some embodiments of the MXene nanomaterial with high SOD enzyme activity described in the present application, the metal atom is a copper atom.

[0008] In a second aspect of the present application, a method for preparing the MXene nanomaterial with high SOD enzyme activity according to the first aspect of the present application is provided, comprising the following steps: mixing Mo2C MXene, an inorganic metal compound, and a solvent to obtain a MXene nanomaterial.

[0009] According to some embodiments of the preparation method described in the present application, the inorganic metal compound includes one or more of CuCl2·3H2O, Cu(NO3)2, FeCl3·6H2O, H2PtCl6·6H2O, ZnCl2 and MnCl2·4H2O.

[0010] According to some embodiments of the preparation method described in the present application, the solvent includes deionized water and / or ethanol.

[0011] According to some embodiments of the preparation method described herein, the mass ratio of the Mo2C MXene to the inorganic metal compound is (10-20):(1-3).

[0012] According to some embodiments of the preparation method described in the present application, the mixing temperature is 20-30° C., and the mixing time is 30-90 min.

[0013] According to some embodiments of the preparation method described in the present application, the preparation method further includes filtering the mixed product and drying the filter cake to obtain the MXene nanomaterial.

[0014] According to some embodiments of the preparation method described in the present application, the drying temperature is 60-80° C., and the drying time is 1-2 hours.

[0015] The third aspect of the present application provides an application of the MXene nanomaterial with high SOD enzyme activity described in the first aspect of the present application or the MXene nanomaterial with high SOD enzyme activity obtained by the preparation method described in the second aspect of the present application in the preparation of drugs for treating ROS-related diseases.

[0016] The beneficial effects of the present application include: the MXene nanomaterial with high SOD enzyme activity described in the present application has high SOD enzyme activity and strong ROS scavenging properties, which can reduce oxidative stress damage; it has the effect of treating ROS-related diseases such as traumatic brain injury (TBI), Parkinson's syndrome (PD), and chronic obstructive pulmonary disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 A is a SEM image of the MXene nanomaterial (Cu SA / MXene) with high SOD enzyme activity described in Example 1 of the present application;

[0018] Figure 1B is a TEM image of the MXene nanomaterial (Cu SA / MXene) with high SOD enzyme activity described in Example 1 of the present application;

[0019] Figure 1 C is a HAADF-STEM image of the MXene nanomaterial (Cu SA / MXene) with high SOD enzyme activity described in Example 1 of the present application;

[0020] Figure 2 This is a graph showing the activity measurement results of the MXene nanomaterial (Cu SA / MXene) with high SOD enzyme activity described in Example 1 of the present application;

[0021] Figure 3 This is a graph showing the results of a study on the SOD-like activity of the MXene nanomaterials having high SOD enzyme activity described in Examples 1-5 of this application;

[0022] Figure 4 This is a graph showing the specific activity study results of the MXene nanomaterial with high SOD enzyme activity described in Example 1 of the present application;

[0023] Figure 5 This is a graph showing the results of a ROS scavenging performance study of the MXene nanomaterial with high SOD enzyme activity described in Example 1 of the present application. DETAILED DESCRIPTION

[0024] The embodiments of the present invention are described in detail below. The examples of the embodiments are exemplary and intended to be used to explain the present invention, but should not be understood as limiting the present invention.

[0025] In the present invention, the terms "one embodiment", "some embodiments", "examples", "specific examples", or "some examples" mean that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic expressions of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art can combine and combine different embodiments or examples described in this specification and the features of different embodiments or examples without contradiction.

[0026] The present invention provides a MXene nanomaterial with high SOD enzyme activity, including Mo2C MXene and metal atoms anchored on the Mo2C MXene.

[0027] Mo2C MXene is a two-dimensional transition metal carbide composed of molybdenum carbide.

[0028] The MXene nanomaterial described in this application is composed of metal atoms anchored in Mo2C MXene, and has superoxide dismutase-like activity and catalase (CAT)-like activity, which can reduce the upregulated level of reactive oxygen species.

[0029] In some embodiments of the present application, the anchoring amount of metal atoms on the Mo2C MXene is 2 wt%-3 wt%, for example, 2 wt%, 2.34 wt%, 2.46 wt%, 2.76 wt%, 2.89 wt%, 3 wt%, etc.

[0030] In some embodiments of the present application, the metal atoms include one or more of copper atoms, iron atoms, platinum atoms, manganese atoms and zinc atoms.

[0031] In some embodiments of the present application, the metal atom is a copper atom.

[0032] This embodiment of the present application also provides a method for preparing the MXene nanomaterial with high SOD enzyme activity described in the first aspect of the present application, comprising the steps of: mixing Mo2C MXene, an inorganic metal compound, and a solvent to obtain the MXene nanomaterial. The preparation method described herein is simple and does not require the addition of an additional reducing agent.

[0033] In some embodiments of the present application, the inorganic metal compound includes one or more of CuCl2·3H2O, Cu(NO3)2, FeCl3·6H2O, H2PtCl6·6H2O, ZnCl2 and MnCl2·4H2O.

[0034] In some embodiments of the present application, the inorganic metal compound includes CuCl2 and / or Cu(NO3)2 and hydrates thereof.

[0035] In some embodiments of the present application, the solvent includes deionized water and / or ethanol.

[0036] In some embodiments of the present application, the mass ratio of the Mo2C MXene to the inorganic metal compound is (10-20):(1-3), for example, 10:1, 10:2, 10:3, 15:1, 15:2, 15:3, 20:1, 20:2, 20:3, etc.

[0037] In some embodiments of the present application, the mixing temperature is 20-30°C, for example, 20°C, 25°C, 28°C, 30°C, etc., and the mixing time is 30-90min, for example, 30min, 35min, 40min, 48min, 53min, 60min, 90min, etc.

[0038] In some embodiments of the present application, the preparation method further includes filtering the mixed product and drying the filter cake to obtain the MXene nanomaterial.

[0039] In some embodiments of the present application, the drying temperature is 60-80°C, for example, 60°C, 65°C, 68°C, 73°C, 76°C, 80°C, etc., and the drying time is 1-2h, for example, 1h, 1.2h, 1.5h, 1.8h, 2h, etc.

[0040] The embodiments of the present application also provide an application of the MXene nanomaterial with high SOD enzyme activity described in the first aspect of the present application or the MXene nanomaterial with high SOD enzyme activity obtained by the preparation method described in the second aspect of the present application in the preparation of drugs for treating ROS-related diseases.

[0041] The technical solution of this application is further described below with reference to specific embodiments.

[0042] Example 1

[0043] A method for preparing a MXene nanomaterial with high SOD enzyme activity comprises the following steps:

[0044] 20 mg of Mo2C MXene, 3 mg of CuCl2·2H2O and 23 mL of deionized water were mixed and stirred at 25°C for 30 min, and then ultrasonicated with argon for 1 h. The resulting mixture was then filtered through a polytetrafluoroethylene (PTFE) membrane under vacuum conditions, and the filter cake was dried at 65°C for 1 h to obtain a MXene nanomaterial with high SOD enzyme activity, in which the anchoring amount of Cu atoms on Mo2C MXene was 2.43 wt% (the Cu loading in Cu SA / MXene was 2.43 wt% quantified by inductively coupled plasma atomic emission spectrometry (ICP-AES), recorded as Cu SA / MXene.

[0045] Example 2

[0046] The preparation method of the MXene nanomaterial with high SOD enzyme activity described in Example 2 is different from that in Example 1 only in that FeCl3·6H2O is used instead of CuCl2·2H2O in the preparation process of the MXene nanomaterial described in Example 2.

[0047] The specific steps include:

[0048] 20 mg of Mo2C MXene, 4.75 mg of FeCl3·6H2O, and 25 mL of deionized water were mixed and stirred at 25°C for 30 min, and then ultrasonicated with argon for 1 h. The resulting mixture was then filtered through a polytetrafluoroethylene (PTFE) membrane under vacuum conditions, and the filter cake was dried at 65°C for 1 h to obtain a MXene nanomaterial with high SOD enzyme activity. The anchoring amount of Fe atoms on Mo2C MXene was 2.43 wt% (the Fe loading in Fe SA / MXene was 2.43 wt% as quantified by inductively coupled plasma atomic emission spectrometry (ICP-AES). This material is denoted as Fe SA / MXene.

[0049] Example 3

[0050] The preparation method of the MXene nanomaterial with high SOD enzyme activity described in Example 3 is different from that in Example 1 only in that H2PtCl6·6H2O is used instead of CuCl2·2H2O in the preparation process of the MXene nanomaterial described in Example 3.

[0051] The specific steps include:

[0052] 20 mg of Mo2C MXene, 9.09 mg of H2PtCl6·6H2O, and 29 mL of deionized water were mixed and stirred at 25°C for 30 min, and then ultrasonicated with argon for 1 h. The resulting mixture was then filtered through a polytetrafluoroethylene (PTFE) membrane under vacuum conditions, and the filter cake was dried at 65°C for 1 h to obtain a MXene nanomaterial with high SOD enzyme activity, in which the anchoring amount of Pt atoms on Mo2C MXene was 2.43 wt% (the Pt loading amount in PtSA / MXene was 2.43 wt% quantified by inductively coupled plasma atomic emission spectrometry (ICP-AES), recorded as Pt SA / MXene.

[0053] Example 4

[0054] The preparation method of the MXene nanomaterial with high SOD enzyme activity described in Example 4 is different from that in Example 1 only in that MnCl2·4H2O is used instead of CuCl2·2H2O in the preparation process of the MXene nanomaterial described in Example 4.

[0055] The specific steps include:

[0056] 20 mg of Mo2C MXene, 3.47 mg of MnCl2·4H2O, and 24 mL of deionized water were mixed and stirred at 25°C for 30 min, and then ultrasonicated with argon for 1 h. The resulting mixture was then filtered through a polytetrafluoroethylene (PTFE) membrane under vacuum conditions, and the filter cake was dried at 65°C for 1 h to obtain a MXene nanomaterial with high SOD enzyme activity. The anchoring amount of Mn atoms on Mo2C MXene was 2.43 wt% (the Mn loading in Mn SA / MXene was 2.43 wt% as quantified by inductively coupled plasma atomic emission spectrometry (ICP-AES), which was recorded as Mn SA / MXene.

[0057] Example 5

[0058] The preparation method of the MXene nanomaterial with high SOD enzyme activity described in Example 5 is different from that in Example 1 only in that ZnCl2 is used instead of CuCl2·2H2O in the preparation process of the MXene nanomaterial described in Example 5.

[0059] The specific steps include:

[0060] 20 mg of Mo2C MXene, 2.39 mg of ZnCl2 and 22 mL of deionized water were mixed and stirred at 25°C for 30 min, ultrasonicated with argon for 1 h, and then filtered through a polytetrafluoroethylene (PTFE) membrane under vacuum conditions. The filter cake was dried at 65°C for 1 h to obtain a MXene nanomaterial with high SOD enzyme activity, in which the anchoring amount of Zn atoms on Mo2C MXene was 2.43 wt% (the Zn loading amount in Zn SA / MXene was 2.43 wt% quantified by inductively coupled plasma atomic emission spectrometry (ICP-AES), which was recorded as Zn SA / MXene.

[0061] 1. The SEM, TEM and HAADF-STEM images of the MXene nanomaterial (Cu SA / MXene) with high SOD enzyme activity described in Example 1 of the present application are as follows: Figure 1 A. Figure 1 B and Figure 1 As shown in C.

[0062] from Figure 1 A and Figure 1 It can be seen from B that the morphology of the MXene nanomaterial with high SOD enzyme activity described in Example 1 of the present application is a lamellar layer.

[0063] from Figure 1 It can be seen from C that the MXene nanomaterial Cu with high SOD enzyme activity described in Example 1 of the present application is in an atomic-level dispersion state on the MXene ( Figure 1 The red circles in C indicate the presence of Cu atoms).

[0064] 2. Activity determination of MXene nanomaterial (Cu SA / MXene) with high SOD enzyme activity described in Example 1 of the present application

[0065] Test method: The MXene nanomaterial and Mo2CMXene with CAT enzyme-like activity prepared in Example 1 of the present application were mixed with 10mM H2O2 solution in a cuvette (the dosage of MXene nanomaterial and Mo2C MXene was 100μg / mL, and Cu SA / MXene catalyzed the decomposition of H2O2 in aqueous solution, resulting in the generation of O2 bubbles, showing significant catalase-like activity). Then, the intensity of the characteristic absorption peak of H2O2 at 240nm in each cuvette was evaluated at 25°C to monitor the consumption of H2O2. The results are as follows: Figure 2 shown.

[0066] from Figure 2 It can be seen that Cu SA / MXene has CAT enzyme-like activity.

[0067] 3 Study on the SOD-like activity of MXene nanomaterials with high SOD enzyme activity described in Examples 1-5 of this application

[0068] Test method: The SOD specific activity of the MXene nanomaterial samples described in Examples 1-5 was determined using the nitroblue tetrazolium (NBT) method.

[0069] O2 ·- Nitroblue tetrazolium (NBT) can be reduced to blue-purple formazan, which has a maximum absorption at 560nm. O2 is detected by detecting the absorption at 560nm ·- O2 ·- The higher the content, the higher the absorption at 560nm.

[0070] First, xanthine (0.25 mM) and xanthine oxidase (0.05 U / mL) were mixed in phosphate buffer (50 mM, pH 7.4) at 25 °C to prepare O2 ·- , then NBT was added and reacted for 10 minutes (6 groups were prepared under the same conditions).

[0071] Add O2 to the above 6 groups respectively ·- The MXene nanomaterials with high SOD enzyme activity described in Examples 1-5 (dosage of 100 μg / mL) and Mo2C MXene (dosage of 100 μg / mL) were added to the solution. After reacting for 10 minutes, the UV-visible light absorption at 560 nm was detected, and the NBT reduction inhibition rate of each sample was calculated.

[0072] The test results are as follows Figure 3 shown.

[0073] from Figure 3 It can be seen that Cu SA / MXene has ultra-high SOD enzyme activity, which is better than Fe SA / MXene, MnSA / MXene, Zn SA / MXene, and Pt SA / MXene.

[0074] 4. Study on the specific activity of MXene nanomaterials with high SOD enzyme activity described in Example 1 of this application

[0075] Test method: O2 ·- With xanthine (0.25mM) and xanthine oxidase (0.05U·mL -1 ) were mixed in phosphate buffer (50 mM, pH 7.4) at 37°C, and then NBT was added and reacted for 10 min (two groups were prepared under the same conditions).

[0076] Cu SA / MXene (100 μg / mL) and MoC MXene (100 μg / mL) were added to the two solutions, respectively. After 10 minutes of reaction, UV-visible absorbance at 560 nm was measured. Under these specific conditions, the amount of nanozyme required to inhibit formazan formation by 50% was defined as 1 unit of SOD-like activity.

[0077] The inhibition rates of nanozyme samples at different concentrations were measured, and the half-inhibitory concentration (IC50) was calculated using the dose-response inhibition function analysis in GraphPad Prism 7 software. Then, the SOD specific activity of nanozyme was calculated using the following formula: SOD SA (U·mg -1 )=1 / (IC50·V), the results are as follows Figure 4 shown.

[0078] from Figure 4 It can be seen that the SOD-like enzyme specific activity of Cu SA / MXene is 29904.3 U / mg.

[0079] 5. Study on the ROS scavenging performance of MXene nanomaterials with high SOD enzyme activity described in Example 1 of this application Test method: H2O2 model active oxygen species determination

[0080] Cultured PC12 cells were seeded into 6-well plates to ensure a cell density of 1.2 × 10 cells per well. 5 After 24 h of incubation, the cells were treated with 25 μg mL -1 or 50 μg mL -1The cells were treated with fresh Cu SA / MXene medium instead of the old medium and incubated for a further 1 hour. H2O2 was then added at a concentration of 200 μM, and the cells were incubated for an additional 6 hours. Finally, the reactive oxygen species probe DCFH-DA was introduced and incubated with the cells for 30 minutes. After washing three times with phosphate-buffered saline (PBS), the cells were imaged under a fluorescence microscope.

[0081] from Figure 5 It can be seen that after treatment with the MXene nanozyme (CuSA / MXene) with high SOD activity described in Example 1 of the present application, the fluorescence intensity is significantly reduced, that is, the MXene nanozyme with high SOD activity described in the present application has a strong ROS scavenging property.

[0082] Although the above embodiments have been shown and described, it is understood that the above embodiments are illustrative and are not to be construed as limitations on the present invention. Changes, modifications, substitutions and variations of the above embodiments by those skilled in the art are all within the scope of protection of the present invention.

Claims

1. A MXene nanomaterial with high SOD enzyme activity, characterized in that: Including Mo2C MXene and metal atoms anchored on Mo2C MXene.

2. The MXene nanomaterial with high SOD enzyme activity according to claim 1, characterized in that The anchoring amount of metal atoms on the Mo2CMXene is 2 wt%-3 wt%.

3. The MXene nanomaterial with high SOD enzyme activity according to claim 1, characterized in that The metal atom includes one or more of copper atom, iron atom, platinum atom, manganese atom and zinc atom, preferably copper atom.

4. The method for preparing the MXene nanomaterial with high SOD enzyme activity according to any one of claims 1 to 3, characterized in that: The method comprises the following steps: mixing Mo2C MXene, an inorganic metal compound and a solvent to obtain a MXene nanomaterial.

5. The method for preparing a MXene nanomaterial having high SOD enzyme activity according to claim 4, characterized in that: The inorganic metal compound includes one or more of CuCl2·2H2O, Cu(NO3)2, FeCl3·6H2O, H2PtCl6·6H2O, ZnCl2 and MnCl2·4H2O; And / or, the solvent includes deionized water and / or ethanol.

6. The method for preparing a MXene nanomaterial having high SOD enzyme activity according to claim 4, characterized in that: The mass ratio of the Mo2C MXene to the inorganic metal compound is (10-20): (1-3).

7. The method for preparing a MXene nanomaterial having high SOD enzyme activity according to claim 4, characterized in that: The mixing temperature is 20-30° C., and the mixing time is 30-90 min.

8. The method for preparing a MXene nanomaterial with high SOD enzyme activity according to claim 4, characterized in that: The preparation method further includes filtering the mixed product and drying the filter cake to obtain the MXene nanomaterial.

9. The method for preparing a MXene nanomaterial having high SOD enzyme activity according to claim 8, characterized in that: The drying temperature is 60-80° C., and the drying time is 1-2 hours.

10. Use of the MXene nanomaterial with high SOD enzyme activity according to any one of claims 1 to 3 or the MXene nanomaterial with high SOD enzyme activity obtained by the preparation method according to any one of claims 4 to 9 in the preparation of drugs for treating ROS-related diseases.

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