Fullerol with moderate hydroxylation degree as well as preparation and application of fullerol

Abstract

The invention belongs to the technical field of nano materials, and relates to fullerol with moderate hydroxylation degree and a preparation method and application thereof. Fullerol with the chemical formula of C60 (OH) n.mH2O is obtained by controlling the mass ratio of fullerene to an alkaline reagent and adopting the processes of grinding, dissolving, concentration, alcohol precipitation and drying, n is 10-15, preferably 12, m is 10-15, preferably 12.5, and the product is a needle-like crystal and forms a stable dispersion liquid in water. Thermogravimetric analysis shows that the fullerol has controllable hydroxyl number and relatively high water solubility, has better ABTS < + > and. OH removal capability compared with low-hydroxylated and high-hydroxylated fullerol, can reduce the ROS level in cells and improve the cell survival rate and migration capability, can be used as an active component of an antioxidant and a cell repair preparation, and has a broad application prospect. The composition is used for skin barrier repair and oxidative stress related disease prevention and treatment.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of nanomaterials and their biomedical applications, and specifically relates to a fullerol with moderate hydroxylation degree and a preparation method thereof, and relates to the application of the fullerol in antioxidation and cell repair. BACKGROUND

[0002] Reactive oxygen species (ROS) are a class of active oxygen-containing molecules produced in the process of biological oxidation metabolism, and play an important role in maintaining cell signal transduction and physiological homeostasis. When ROS accumulates excessively, the cell is in an oxidative stress state, which may lead to lipid peroxidation, protein denaturation and DNA damage, thereby inducing inflammatory response and related pathological changes.

[0003] Fullerenes (C 60 ) have a unique cage-like conjugated structure and high electron affinity, and can effectively scavenge various free radicals and ROS through electron transfer or addition reaction, showing great application potential in antioxidation and cell protection. However, intrinsic fullerenes are highly hydrophobic and can only be dissolved in a few organic solvents, and have very low solubility in water, which limits their practical application in water-phase systems such as biological medicine and skin repair.

[0004] To improve the water solubility and biocompatibility of fullerenes, the prior art usually prepares fullerols by introducing hydrophilic groups such as hydroxyl groups on the surface of fullerenes. Hydroxylation can adjust the charge and aggregation behavior of the surface of fullerenes, enhance the interaction with biological macromolecules such as proteins and cell membranes, and to some extent, improve the antioxidant and cell repair activities.

[0005] However, the hydroxylation level of fullerols has an important influence on the material properties: Firstly, too low hydroxylation level will result in insufficient dispersibility and stability of the material in water environment, which is not conducive to in vitro and in vivo applications; Secondly, too high hydroxylation level may destroy the π-conjugated skeleton structure of C 60 , weaken its electron transfer ability with free radicals, and thus affect the antioxidant efficiency; Thirdly, in the existing preparation methods, the number of hydroxyl groups is often difficult to accurately control, and the process flow is relatively complex, and some methods need to involve organic solvents, phase transfer catalysts or strong oxidants, which is not conducive to large-scale preparation and safe use.

[0006] In addition, the existing literature lacks systematic quantitative research and clear conclusions on the differences and optimal balance relationship of different hydroxylation degree fullerols in terms of in vitro free radical scavenging ability, intracellular antioxidant performance and cell repair capacity.

[0007] Therefore, how to establish a preparation method with simple process, easy-to-control conditions, and stable obtainment of fullerols with moderate hydroxylation degree, and systematically verify the comprehensive performance of the fullerols in antioxidant and cell repair is a technical problem to be solved in the field. SUMMARY

[0008] The present application aims to solve the problems in the prior art, such as difficulty in accurately controlling the hydroxylation degree of fullerols, difficulty in balancing water solubility and antioxidant performance, and complex preparation process, and provides a fullerol with moderate hydroxylation degree and a preparation method thereof, and further provides applications of the fullerol in antioxidant and cell repair. By controlling the number of hydroxyl groups grafted on the surface of fullerene, the fullerol is in the "moderate hydroxylation" interval balancing water solubility and antioxidant activity, so that the material with comprehensive performance better than low-hydroxylated and high-hydroxylated fullerols is obtained.

[0009] To achieve the above-mentioned purpose, the present application adopts the following technical solutions: In a first aspect, the present application provides a fullerol with moderate hydroxylation degree.

[0010] The chemical formula of the fullerol is C 60 (OH) n ·mH2O, wherein n is 10-15, preferably 12, and m is 10-15, preferably 12.5. The fullerol can form a transparent or translucent stable dispersion in water, and has a needle-like crystal morphology and uniform distribution under a scanning electron microscope.

[0011] Preferably, through thermogravimetric analysis, it can be observed that the fullerol has obvious three-stage mass loss in three temperature zones of 30-186 ℃, 186-381 ℃, and higher than 381 ℃, corresponding to removal of combined water, removal of hydroxyl groups, and decomposition of C 60 skeleton. Based on the mass proportion relationship of the weight loss curve, the number of hydroxyl groups n and the hydration number m can be calculated to confirm the structure parameters of the fullerol.

[0012] In a second aspect, the present application provides a preparation method of the above-mentioned fullerol with moderate hydroxylation degree.

[0013] The preparation method comprises the following steps: (1) Fullerene and an alkaline reagent are mixed and ground at a mass ratio of 1:(3-5) for 10-20 minutes until the color of the material changes from black to yellow-brown, to obtain a uniform mixture; (2) Deionized water is added to the mixture, and after being fully dissolved under stirring, a transparent filtrate is obtained by filtration; (3) The obtained filtrate is heated and concentrated at 25-50 ℃ to obtain a concentrated solution; (4) adding an alcohol solvent to the concentrated solution to wash and precipitate, and collecting the precipitate by centrifugation, and repeating the washing-precipitation-centrifugation steps as needed; (5) vacuum drying the precipitate at 60 ℃ to obtain the fullerol with moderate degree of hydroxylation.

[0014] Preferably, the alkaline agent is sodium hydroxide, the alcohol solvent is methanol, the mass ratio of fullerene to sodium hydroxide is preferably 1:4, and the grinding time is preferably about 15 minutes. By using the above preferred process conditions, C 60 (OH) 12 ·12.5H2O.

[0015] In a third aspect, the present application provides the use of the above-mentioned fullerol with moderate degree of hydroxylation in antioxidation and cell repair.

[0016] The fullerol can effectively scavenge free radicals such as ABTS + and ·OH in vitro, and reduce intracellular ROS levels in a LPS-induced cell inflammation model, thereby achieving antioxidant protection. Meanwhile, the fullerol has good cell compatibility in a suitable concentration range, can improve cell survival rate, and significantly promotes cell migration and damage repair in a cell scratch model, and can be used as an active ingredient to prepare an antioxidant protection agent, a cell repair preparation, and a skin barrier repair product.

[0017] In a fourth aspect, the present application further provides an antioxidant or cell repair composition.

[0018] The composition comprises the above-mentioned fullerol with moderate degree of hydroxylation and a pharmaceutically or dermatologically acceptable carrier, and can be prepared into a drug form such as an external preparation, an injection, or a dressing product for use in improving skin barrier function and / or preventing or relieving oxidative stress-related diseases or injuries.

[0019] Compared with the prior art, the present application has the following beneficial effects: (1) The present application can stably prepare a fullerol with moderate degree of hydroxylation under mild conditions by optimizing the combination of the ratio of fullerene-alkaline system and the steps of grinding, dissolving, concentrating, precipitating, and drying, and the process flow is simple and easy to scale up, thereby solving the problem of difficulty in accurately controlling the number of hydroxyl groups in the prior art.

[0020] (2) The number of hydroxyl groups n of the fullerol obtained by the present application is limited to a moderate range of 10-15, which not only significantly improves the solubility and dispersion stability of the material in an aqueous system, but also avoids excessive hydroxylation of C 60The adverse effects of the conjugated skeleton and free radical scavenging ability were mitigated, achieving a favorable balance between water solubility and antioxidant properties. Thermogravimetric analysis results and data in Table 1 show that the mass loss of this hydroxylated fullerol during the water and hydroxyl group removal stages is highly consistent with theoretical calculations, indicating that the structural parameters are clearly defined and controllable.

[0021] (3) In the examples, low-hydroxylated fullerols (e.g., C) 60 (OH)8·7H2O), hydroxylated fullerol (C 60 (OH) 12 ·12.5H2O) and highly hydroxylated fullerols (e.g., C 60 (OH) 43 In vitro ABTS (·8.5H2O) + Results of the · and ·OH removal experiments (see) Figure 3 , Figure 4 The results show that, at the same concentration, the free radical scavenging rate of hydroxylated fullerol is significantly higher, proving that the present invention achieves an optimal balance between the number of hydroxyl grafts and antioxidant performance.

[0022] (4) Results of cell-level experiments (see) Figures 5 to 8 The results show that, within a suitable concentration range, the hydroxylated fullerol of this invention exhibits low cytotoxicity, effectively reduces intracellular ROS levels in the LPS-induced model, and promotes cell migration and scratch healing, demonstrating good cell repair potential and biosafety. This fullerol can be used as a single active ingredient or formulated with pharmaceutically or dermatologically acceptable carriers to create various dosage forms such as antioxidant or cell repair agents, showing broad application prospects. Attached Figure Description

[0023] Figure 1 This is a comparison of thermogravimetric analysis (TGA) curves of fullerols with different degrees of hydroxylation. Figure 1 (a) is the thermogravimetric curve of highly hydroxylated fullerol. Figure 1 (b) The thermogravimetric curve of fullerol with a moderate degree of hydroxylation. Figure 1 (c) shows the thermogravimetric curves of low-hydroxylated fullerols, used to characterize the mass loss characteristics of each sample in different temperature ranges and the structural parameters estimated therefrom.

[0024] Figure 2 This is a scanning electron microscope (SEM) image of fullerol with a moderate degree of hydroxylation according to the present invention, used to show the needle-like crystal morphology and uniformity of distribution of the product.

[0025] Figure 3 ABTS for fullerols with different degrees of hydroxylation +• Curves showing the free radical scavenging rate as a function of concentration are used to compare the in vitro antioxidant capacity of highly hydroxylated, moderately hydroxylated, and lowly hydroxylated fullerols.

[0026] Figure 4 The graph shows the scavenging rate of fullerols with different degrees of hydroxylation over time, used to compare the effects of different hydroxyl grafting levels on the scavenging rate and efficiency of hydroxyl radicals.

[0027] Figure 5 This image shows the fluorescence imaging and quantitative results of the antioxidant effect of fullerol with a moderate degree of hydroxylation in NCM-460 cells. Figure 5 (a) Laser confocal fluorescence imaging of the fullerol treatment group and the control group with different concentrations and appropriate degrees of hydroxylation. Figure 5 (b) The average fluorescence intensity of each group is statistically analyzed using ImageJ software, which reflects the changes in intracellular ROS levels.

[0028] Figure 6 The statistical graph shows the survival rate of NCM-460 cells treated with fullerol at different concentrations with appropriate degrees of hydroxylation, which is used to characterize the cytotoxicity and safe dosage range of this material.

[0029] Figure 7 These are micrographs showing the morphological changes of the cell scratch region at different time points (0 h, 12 h, 24 h and 36 h) under the action of fullerol with a moderate degree of hydroxylation at 0.2 mg / mL, used to evaluate cell migration and scratch closure process.

[0030] Figure 8 For the corresponding Figure 7 The statistical graph of cell scratch area shown in the experiment was used to quantitatively analyze the effect of fullerol with moderate hydroxylation on cell migration rate and scratch healing degree. Detailed Implementation

[0031] The present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Those skilled in the art should understand that the following embodiments are for illustrative purposes only and are not intended to limit the scope of protection of the present invention. Equivalent substitutions or adjustments made to specific process parameters, raw material ratios, and testing methods without departing from the spirit and substance of the present invention should be considered to fall within the scope of protection of the present invention.

[0032] Example 1: Preparation of fullerols with moderate hydroxylation 72 mg of fullerene C 60Mixed with 240 mg of sodium hydroxide, placed in a mortar and ground thoroughly for about 15 min until the mixture gradually changed from black to yellowish brown, indicating that the initial hydroxylation reaction of the fullerene under alkaline conditions had occurred, obtaining a uniform mixture. Subsequently, 50 mL of deionized water was added to the mixture, and the reaction was continued under stirring for about 10 min to fully dissolve the solid and form a uniform solution. The obtained solution was filtered, and the insoluble matter was discarded to obtain a transparent filtrate.

[0033] The above filtrate was heated and concentrated to about 5 mL at 25-50 °C to obtain a concentrated solution. Excess methanol was added to the concentrated solution while stirring to precipitate the target product. The precipitate was collected by centrifugation, and the precipitate was repeatedly washed with methanol and centrifuged. The washing and centrifugation steps were repeated three times to remove residual inorganic salts and unreacted raw materials. Finally, the precipitate was placed in a 60 °C vacuum drying oven for drying until the mass was constant, obtaining a fullerol with moderate degree of hydroxylation (hereinafter referred to as "moderate hydroxylation fullerol").

[0034] The obtained moderate hydroxylation fullerol was subjected to thermogravimetric analysis (TGA), and the results are shown in Figure 1 (b). As can be seen, the mass loss of the sample mainly occurs in three temperature intervals: in the range of 30-186 °C, the mass of the sample decreases by about 19.72% (y1), corresponding to the gradual removal of water of hydration; in the range of 186-381 °C, the mass further decreases by about 17.66% (y2), mainly due to the removal of hydroxyl groups attached to the C 60 core; when the temperature is higher than 381 °C, the mass continues to decrease by about 62.62% (y3 = 1-y1-y2), corresponding to the decomposition of the C 60 skeleton. Combined with the above mass loss ratio and referring to the theoretical composition of C 60 (OH) x ·mH2O, it can be calculated that the structure of the sample is C 60 (OH) 12 ·12.5H2O.

[0035] The scanning electron microscope (SEM) image is shown in Figure 2 , and the moderate hydroxylation fullerol has an elongated needle-like crystal morphology, with uniform particle size and good dispersion, indicating that the process of the present application can produce products with regular morphology and uniform distribution.

[0036] To compare the structural characteristics of fullerenols with different degrees of hydroxylation, the thermogravimetric data and structural parameters shown in Table 1 were obtained.

[0037] Table 1 Example of structural parameters of fullerenols with different degrees of hydroxylation estimated based on TGA data As can be seen from Table 1, the hydroxyl number (x = 12) of the mesohydroxylated fullerol is between the high-hydroxylated and low-hydroxylated samples, and the hydration number (m = 12.5) verifies that the preparation process of the present application indeed obtains a fullerol with moderate degree of hydroxylation.

[0038] Preparation of low-hydroxylated fullerol 2 g of sodium hydroxide was dissolved in 20 mL of deionized water to prepare a sodium hydroxide aqueous solution. 50 mg of fullerene was dissolved in 50 mL of benzene to prepare a fullerene organic phase solution. 10 mg of cetyltrimethylammonium bromide (CTAB) and 5 drops of 30% hydrogen peroxide solution were added to the above sodium hydroxide solution, and the fullerene benzene solution was added dropwise into the alkaline aqueous phase under vigorous stirring, and the stirring was continued until the organic layer became colorless.

[0039] After the reaction was completed, the red-brown aqueous phase was separated, and ethanol was added to the aqueous phase, and the product was precipitated by standing. The obtained precipitate was centrifuged and the supernatant was discarded, and the ethanol precipitation and centrifugation steps were repeated three times, and finally dried at 60 ℃ for 5 h to obtain low-hydroxylated fullerol.

[0040] Thermogravimetric analysis was performed on the obtained sample, and the results are shown in Figure 1 (c). According to the mass loss curve and referring to Table 1, the product structure can be deduced as C 60 (OH)8·7H2O, i.e., a lower degree of hydroxylation.

[0041] Preparation of high-hydroxylated fullerol 99.7 mg of fullerene, 50 mL of toluene, 10 mL of 30% hydrogen peroxide solution and 0.8 mL of tetrabutylammonium hydroxide were mixed and placed in a reaction bottle, and stirred at 60 ℃ at a speed of 700 r / min for 1 h. After the reaction was completed, the layers were separated by standing, and the lower yellow aqueous phase was separated.

[0042] Isopropyl alcohol, diethyl ether and n-hexane were mixed in a volume ratio of 7:5:5 as a precipitant, and the above yellow aqueous phase was added to the precipitant and divided into 4 parts, and centrifuged at 8000 r / min for 5 min, and the yellow precipitate was collected. The precipitate was washed with diethyl ether and centrifuged again, and the step was repeated several times, and the precipitate was dried to obtain high-hydroxylated fullerol.

[0043] Thermogravimetric analysis results are shown in Figure 1 (a). According to the mass loss curve and referring to Table 1, the product structure can be deduced as C 60 (OH) 43 ·8.5H2O, indicating a higher degree of hydroxylation.

[0044] Example 2 Comparison of antioxidant performance in vitro of different hydroxylated fullerols (1) ABTS + · Assay of clearance rate ABTS radical scavenging method was used to evaluate the antioxidant activity in vitro of different hydroxylated fullerols. The high, medium and low hydroxylated fullerols were respectively prepared into solutions with a mass concentration of 1, 0.5, 0.25, 0.125 mg / mL using deionized water. ABTS + · working solution (ABTS was fully reacted with oxidant in advance to generate stable ABTS + · solution).

[0045] In a 96-well plate, 50 μL of sample solution of different concentrations was mixed with 150 μL of ABTS + · working solution, and the absorbance of each well was measured at 734 nm after reaction at room temperature for 15 min in the dark. Deionized water was used as a control instead of sample solution, and the radical scavenging rate was calculated according to the change in absorbance.

[0046] The results are shown in Figure 3 , which show that the ABTS + · clearance rate of the medium hydroxylated fullerol is higher than that of the high and low hydroxylated fullerols at all concentrations, and gradually increases with increasing concentration, showing good concentration dependence.

[0047] (2) Assay of ·OH clearance rate Hydroxyl radicals (·OH) were generated using a Fenton reaction system, and 3,3',5,5'-tetramethylbenzidine (TMB) was used as a chromogenic agent to evaluate the ·OH scavenging capacity of different samples by measuring the change in absorbance.

[0048] The high, medium and low hydroxylated fullerols were respectively prepared into solutions with a mass concentration of 1, 0.5, 0.25, 0.125 mg / mL using deionized water. In a 96-well plate, 50 μL of sample solution, 50 μL of FeSO4 solution and 50 μL of 0.12% H2O2 solution were sequentially added, and 50 μL of TMB solution was finally added to start the reaction. The absorbance was measured at 652 nm after reaction at room temperature for 20 min in the dark, and a system without sample solution under the same conditions was used as a control.

[0049] As shown in Figure 4 , the ·OH clearance rate of the medium hydroxylated fullerol is significantly higher than that of the high and low hydroxylated fullerols at the same concentration, also showing concentration dependence. This result is consistent with the ABTS +The scavenging experiments were consistent, further demonstrating that hydroxylated fullerol has comprehensive advantages in free radical scavenging.

[0050] Example 3: Intracellular antioxidant properties of hydroxylated fullerol Inoculate NCM460 cell suspension (cell density approximately 2 × 10⁶ cells / mL) into glass-bottomed cell culture dishes. -5 Cells were incubated in serum-containing medium for 24 h to allow them to adhere and reach an appropriate density. The original medium was then discarded, and both the control and experimental groups were replaced with 1 mL of basal medium containing 1 μg / mL LPS to establish a cellular inflammation model.

[0051] The experimental groups were further incubated with basal medium containing different concentrations of hydroxylated fullerol, namely 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL; the control group was incubated with basal medium without fullerol. All groups were incubated for another 24 h.

[0052] After incubation, 2',7'-dichlorofluorescein diacetate (DCFHDA) was diluted according to the manufacturer's instructions and added to each group's culture dish for in situ loading. The cells were incubated at 37 °C for a certain period to allow the probes to enter the cells and be hydrolyzed by esterases. Cells were gently washed with PBS to remove any probes that had not entered the cells. Subsequently, fluorescence images of each group of cells were acquired using a laser confocal microscope at an excitation wavelength of 488 nm and an emission wavelength of 525 nm.

[0053] ImageJ software was used to perform quantitative analysis of the average fluorescence intensity of each group of images, and the results are as follows: Figure 5 As shown. Figure 5 (a) Confocal fluorescence images of the treatment group and the control group at different concentrations of hydroxylated fullerol. Figure 5 (b) is a statistical graph of the corresponding average fluorescence intensity. It can be seen that the control group has the highest fluorescence intensity, indicating a high level of intracellular ROS; the average fluorescence intensity of all experimental groups is significantly lower than that of the control group, indicating that hydroxylated fullerol can effectively scavenge intracellular ROS.

[0054] With increasing concentrations of hydroxylated fullerol, the fluorescence intensity generally decreased, indicating enhanced ROS scavenging effect. The 0.25 mg / mL group showed the most significant decrease, exhibiting a highly significant difference compared to the control group (P < 0.001). The fluorescence intensities of the 0.063 mg / mL and 0.125 mg / mL groups were similar, both significantly lower than the control group (P < 0.01), but slightly higher than the 0.25 mg / mL group. These results demonstrate that hydroxylated fullerol possesses good intracellular antioxidant capacity across a wide concentration range, with the effect being even better near 0.25 mg / mL.

[0055] Example 4: Cytotoxicity test of hydroxylated fullerol The toxicity of hydroxylated fullerol to NCM460 cells was assessed using the CCK8 assay.

[0056] Sample group: NCM460 cells were loaded at a rate of 2 × 10⁻⁶. -4 Seeds were planted in 96-well plates at a density of 1000 cells / well and seeded with serum-containing DMEM. ++ The culture medium was used for 24 h. After discarding the medium, DMEM solution with different concentrations of hydroxylated fullerol (0.125, 0.25, 0.5, and 1.0 mg / mL) was added, and each group was incubated for another 24 h. Then, DMEM solution containing 10% CCK8 was added, and incubation was continued for 1 h. The absorbance A was measured at 450 nm using a microplate reader. s .

[0057] Control group: Except for using DMEM instead of fullerol solution, all other conditions were the same as the sample group. The absorbance at 450 nm was recorded as A. c .

[0058] Sample blank group: No cells were seeded; only hydroxylated fullerol DMEM solution and CCK8 reagent were added to the wells. All other conditions were the same as the sample group. The absorbance was recorded as A. b .

[0059] Control group: No cells were seeded; only DMEM and CCK8 reagent were added. All other conditions were the same as the sample group. The absorbance was recorded as A. bc .

[0060] Three parallel wells were set up for each concentration, and cell viability was calculated using the following formula: Cell viability (%) = [(A s -A b ) / (A c -A bc )]×100% The results are as follows Figure 6 As shown, when the concentration of hydroxylated fullerol was below 0.25 mg / mL, the cell viability was approximately 86.86%, showing only a slight decrease, and no significant cytotoxicity was observed. With further increases in concentration, cell viability decreased, but remained within an acceptable range under experimental conditions. In summary, hydroxylated fullerol exhibits good cell compatibility near its effective antioxidant concentration.

[0061] Example 5: Cell scratch repair experiment with hydroxylated fullerol The cell repair and migration-promoting capabilities of hydroxylated fullerol were evaluated using a cell scratch model.

[0062] NCM460 cells were seeded in culture dishes and cultured until the monolayer cells were confluent. A straight line scratch was gently made on the cell monolayer using a sterile gun tip, and after the detached cells were removed by washing with PBS, the medium containing 0.2 mg / mL C 60 (OH) 12 The same medium without fullerenol was added to the control group. The microscopic morphology of the scratch area was recorded at 0 h, 12 h, 24 h and 36 h, respectively, and the results are shown in Figure 7

[0063] The ImageJ software was used to quantitatively analyze the area of the scratch area at each time point, and the remaining proportion relative to the initial scratch area was calculated, and the results are shown in Figure 8 Compared with the control group, the medium-hydroxylated fullerenol treatment group showed a faster trend of scratch closure throughout the observation period: the scratch width was significantly reduced at 12 h; the cell migration to the blank area was more intensive at 24 h; and the scratch area was mostly covered by new cells at 36 h, with a significantly better healing degree than the control group. This result shows that medium-hydroxylated fullerenol not only has antioxidant effect, but also can promote cell migration and damage repair, and has good cell repair potential.

[0064] The above examples are only specific descriptions of preferred embodiments of the present application and are not intended to limit the present application. For those of ordinary skill in the art, various equivalent substitutions or modifications of the specific conditions, sources of raw materials and experimental methods of the above examples can be made without departing from the spirit and essence of the present application, and these equivalent substitutions or modifications shall fall within the protection scope of the appended claims of the present application.

[0065] Industrial applicability The medium-hydroxylated fullerenol described in the present application uses common chemical raw materials such as fullerene, alkaline reagent and water-alcohol system, and can be prepared by conventional unit operations such as grinding, dissolution, concentration, alcohol precipitation and drying. The reaction temperature is low and the conditions are mild, without the need to use high-pressure equipment or strong oxidizing agents, which is suitable for the existing fine chemical or pharmaceutical auxiliary production line to be scaled up. The obtained fullerenol has clear structure parameters and good batch stability, which is convenient for establishing quality control standards. This material can be used as an antioxidant and cell repair active ingredient, and after being compatible with a pharmaceutically or dermatologically acceptable carrier, it can be further developed into skin care products, pharmaceutical preparations or medical dressings, etc., and has good industrial application prospect.​

Claims

1. A fullerol with a moderate degree of hydroxylation, characterized in that, The chemical formula of the fullerol is C 60 (OH) n ·mH2O, where n is 10-15 and m is 10-15; the fullerol can form a transparent or translucent stable dispersion in water.

2. The fullerol with a suitable degree of hydroxylation according to claim 1, characterized in that, With n = 12 and m = 12.5, the fullerol exhibits a needle-like crystal morphology under a scanning electron microscope, with uniform distribution. Its thermogravimetric analysis curves show the following characteristics in three temperature ranges: 30–186 °C, 186–381 °C, and above 381 °C: removal of bound water, removal of hydroxyl groups, and removal of C... 60 Three-stage mass loss in skeleton decomposition.

3. A method for preparing fullerols with a suitable degree of hydroxylation as described in claim 1 or 2, characterized in that, Includes the following steps: (1) Mixing and grinding: Mix fullerene and alkaline reagent at a mass ratio of 1:(3-5) and grind for 10-20 minutes until the material color gradually changes from black to yellowish-brown to obtain a uniform mixture; (2) Dissolution and filtration: Add deionized water to the mixture, and after stirring, filter to obtain a transparent filtrate; (3) Concentration: The filtrate is heated and concentrated at 25-50 °C to obtain a concentrated solution; (4) Precipitation and separation: Add an alcohol solvent to the concentrate to precipitate the precipitate, collect the precipitate by centrifugation, and optionally wash and centrifuge 2 to 4 times with the alcohol solvent. (5) Drying: The precipitate obtained in step (4) is dried under vacuum at 60 °C to obtain fullerol with a moderate degree of hydroxylation.

4. The preparation method according to claim 3, characterized in that, The alkaline reagent is sodium hydroxide, and the alcohol solvent is methanol.

5. The preparation method according to claim 3 or 4, characterized in that, The mass ratio of fullerene to alkaline reagent is 1:4, and the grinding time is approximately 15 minutes.

6. Fullerol with a suitable degree of hydroxylation prepared by the preparation method according to any one of claims 3 to 5.

7. The use of fullerols with a suitable degree of hydroxylation as described in any one of claims 1, 2, or 6 in the preparation of antioxidant agents, characterized in that, The antioxidant is used to scavenge reactive oxygen species and / or free radicals in vitro or in cells, thereby reducing oxidative stress levels.

8. The use of fullerol with a suitable degree of hydroxylation as described in any one of claims 1, 2, or 6 in the preparation of cell repair agents, characterized in that, The cell repair agent is used to promote cell migration, improve cell survival rate and / or accelerate tissue damage healing, especially to improve skin barrier function.

9. An antioxidant or cell repair composition, characterized in that, The composition comprises a moderately hydroxylated fullerol as described in any one of claims 1, 2 or 6, and a pharmaceutically or dermatologically acceptable carrier.

10. The composition according to claim 9, characterized in that, The composition is formulated as at least one of a topical preparation, an injection, or a dressing for improving skin barrier function and / or preventing or alleviating oxidative stress-related diseases or injuries.

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