Synthesis method of fullerene nano material with glycolipid metabolism regulation capability

Abstract

The invention discloses a synthesis method of a fullerene nano material with glycolipid metabolism regulation capability, and belongs to the technical field of material science and biomedical engineering. The fullerene and the metformin are combined through chemical bonds, the strong oxidation resistance of the fullerene and the hypoglycemic effect of the metformin are exerted at the same time, the synergistic treatment effect is formed, and the drug composition is superior to a single drug; active oxygen in cells can be efficiently removed, oxidative stress is effectively relieved, and the functions of pancreatic beta cells are protected; through structural modification, the problem of low bioavailability caused by hydrophobicity of fullerene and hydrophilicity of metformin is solved, and the solubility and absorption efficiency of the medicine are improved; in conclusion, through chemical combination of the fullerene and the metformin, a dual-mechanism synergistic effect of oxidation resistance and blood sugar reduction is achieved, the treatment effect is remarkably improved, meanwhile, good safety and biocompatibility are achieved, and important application value is achieved in the field of treatment of the type II diabetes mellitus.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of material science and biomedical engineering, and in particular, relates to a synthesis method of a fullerene nanomaterial with glycolipid metabolism regulation ability. BACKGROUND

[0002] The etiology of type Ⅱ diabetes mellitus (T2DM) is complex, but more and more evidence shows that excessive reactive oxygen species (ROS) can induce oxidative stress to affect glycolipid metabolism, leading to insulin resistance and damage to islet beta cells, and promote the development of a series of complications. The unique antioxidant property of fullerene and its derivatives makes it attract much attention in the field of biomedicine. There are data showing that insulin resistance therapeutic agents against oxidative stress have been identified, such as anthocyanins, a-thioctic acid, vitamin E, astaxanthin, flavanol, and other antioxidants. However, due to its limited antioxidant capacity, low bioavailability, poor acid-base stability, and other reasons, its antioxidant effect is not ideal. Therefore, the antioxidant property of fullerene can be used to develop new antioxidants for the treatment of type Ⅱ diabetes mellitus and its complications. However, the hydrophobicity of fullerene itself limits its application in biomedicine, and it can be appropriately functionalized with polar functional groups to produce hydrophilic derivatives to improve solubility. In the reported literature, carboxylated fullerene, polyhydroxylated adduct, amino fullerene, and adducts of fullerene with amino acids, sugars, peptides, and proteins are the most common methods for synthesizing water-soluble fullerene derivatives.

[0003] Metformin can effectively reduce blood glucose in patients and is used as a first-line therapeutic drug for type Ⅱ diabetes mellitus, which has been clinically applied for more than half a century. The hydrophilicity of metformin hinders its ability to pass through the cell membrane by rapid passive diffusion, resulting in slow absorption. The hydrophilicity of metformin can be modified to improve the permeability and passive absorption of highly water-soluble metformin, but it does not seem simple to rationally modify the structure. The simple structural features of metformin may limit structural modification, thereby making it difficult to study the structure-activity relationship of metformin. Fullerene is known as a “free radical sponge” due to its excellent free radical quenching ability and can be used to treat various diseases such as diabetes caused by oxidative stress, and is a promising antioxidant. Carboxylated fullerene is designed and synthesized to increase its water solubility due to its hydrophobicity which is not conducive to its application in the field of biomedicine. In addition, metformin is also a first-line drug for reducing blood glucose. Metformin is easily soluble in water but insoluble in fat, which leads to poor oral absorption and low bioavailability, and cannot prevent the progression of diabetic complications, all of which limit the application of metformin. Therefore, it is urgent to solve the above problems and obtain a new material that can treat and regulate glycolipid metabolism from multiple mechanisms to meet the higher demand in the technical field of treating type Ⅱ diabetes mellitus. SUMMARY

[0004] The present application aims to overcome the defects of the prior art, and provides a synthesis method of fullerene nanomaterials with glycolipid metabolism regulation ability.

[0005] The object of the present application can be achieved by the following technical solutions: The synthesis method of fullerene nanomaterials with glycolipid metabolism regulation ability comprises the following steps: Step S1, put fullerene into anhydrous N,N-dimethylformamide (DMF) and ultrasonically shake for 20-30 min to uniformly disperse the fullerene, to obtain a fullerene dispersion liquid, then add reaction raw materials thiomalic acid (MSA) and initiator dicumyl peroxide to a three-necked flask, and at the same time, add the fullerene dispersion liquid under a nitrogen atmosphere; heat to 140 DEG C, and keep the temperature for 20 h to complete the reaction, then perform post-treatment to obtain carboxylated fullerene; Step S2, add carboxylated fullerene, anhydrous N,N-dimethylformamide (DMF) and thionyl chloride to a three-necked flask, and after acyl chloride reaction at 40 DEG C for 20 h, the reaction is completed, then perform post-treatment to obtain acyl chloride fullerene; Step S3, mix the acyl chloride fullerene with anhydrous N,N-dimethylformamide, ultrasonically shake for 20-30 min to uniformly disperse the fullerene, to obtain an acyl chloride fullerene dispersion liquid, then add freshly prepared free metformin, anhydrous N,N-dimethylformamide and triethylamine to a three-necked flask, stir and mix at room temperature for 1-2 h, then add the acyl chloride fullerene dispersion liquid, heat to 120 DEG C, and keep the temperature for 72 h to complete the reaction, then perform post-treatment to obtain fullerene nanomaterials with glycolipid metabolism regulation ability.

[0006] As a further technical solution, the ratio of the use amounts of fullerene, anhydrous N,N-dimethylformamide, thiomalic acid and dicumyl peroxide in step S1 is 0.2 g:30 mL:3.0 g:2.7 g.

[0007] As a further technical solution, the ratio of the use amounts of carboxylated fullerene, anhydrous N,N-dimethylformamide and thionyl chloride in step S2 is 0.2 g:5 mL:10 mL.

[0008] As a further technical solution, the ratio of the use amounts of freshly prepared metformin, anhydrous N,N-dimethylformamide and triethylamine in step S3 is 0.2 g:23 mL:3.0 g:5 mL.

[0009] As a further technical solution, the operation of post-processing in step S1 is: cooling to room temperature, centrifuging for 10-15 min at 6000-8000 rpm to remove unreacted fullerenes, and then dialyzing the supernatant with deionized water and ethanol three times, respectively, and then spin-drying the suspension in the dialysis bag.

[0010] As a further technical solution, the operation of post-processing in step S2 is: standing for 12 h to precipitate, centrifuging for 10-15 min at 8000-10000 rpm to discard the supernatant and then washing the precipitate with a small amount of tetrahydrofuran, and then drying the precipitate at 50-60 ℃ for 12-24 h.

[0011] As a further technical solution, the operation of post-processing in step S3 is: dialyzing with deionized water and ethanol three times, respectively, after cooling to room temperature, and then drying at 40-50 ℃ under reduced pressure for 24-48 h.

[0012] In step S1 of the present application, thiomalic acid reacts with fullerenes through an ene click reaction under the catalysis of dicumyl peroxide, and a carboxyl group is introduced into the fullerenes skeleton. In step S2, the introduced carboxyl group is acylated to form an acyl chloride group. In step S3, the amino group in metformin reacts with the acyl chloride group to form an amide under the catalysis of triethylamine, and finally the fullerenes nanomaterial with the ability to regulate glycolipid metabolism is obtained.

[0013] The beneficial effects of the present application are: 1. The present application combines fullerenes and metformin through chemical bonds, and simultaneously exerts the strong antioxidant property of fullerenes and the hypoglycemic effect of metformin, forming a synergistic therapeutic effect, which is superior to single drug; 2. The fullerenes derivative of the present application can efficiently scavenge intracellular reactive oxygen species (ROS), effectively alleviate oxidative stress, and protect the function of pancreatic beta cells, which is superior to traditional antioxidants and metformin alone; 3. The present application solves the problem of low bioavailability caused by the hydrophobicity of fullerenes and the hydrophilicity of metformin through structural modification, and improves the solubility and absorption efficiency of the drug; 4. The present application significantly improves the glucose consumption capacity of cells, and the effect is superior to metformin at a therapeutic concentration, especially for cells damaged by oxidative stress; In summary, the present application realizes the synergistic effect of antioxidant and hypoglycemic mechanisms through the chemical combination of fullerenes and metformin, significantly improves the therapeutic effect, and at the same time has good safety and biocompatibility, which has important application value in the field of treating type II diabetes. BRIEF DESCRIPTION OF DRAWINGS

[0014] The present application will be further described below with reference to the accompanying drawings.

[0015] Figure 1 The preparation flow chart of the fullerene nanomaterial with the ability of regulating glycolipid metabolism for the embodiment of the present application; Figure 2 The preparation flow chart of step S1 of the embodiment of the present application; Figure 3 The preparation flow chart of steps S2 and S3 of the embodiment of the present application; Figure 4 The Fourier transform infrared spectrum of the fullerene, carboxylated fullerene, metformin and fullerene nanomaterial with the ability of regulating glycolipid metabolism in the embodiment 1 of the present application; Figure 5 The SEM image of the fullerene, carboxylated fullerene and fullerene nanomaterial with the ability of regulating glycolipid metabolism in the embodiment 1 of the present application; Figure 6 The fluorescence emission spectrum of the carboxylated fullerene and fullerene nanomaterial with the ability of regulating glycolipid metabolism in ethanol in the embodiment 1 of the present application; Figure 7 The bar chart of the islet β cell toxicity test using different concentrations of the fullerene nanomaterial with the ability of regulating glycolipid metabolism in the embodiment 1 of the present application; Figure 8 The fluorescence microscopic image of the determination of the ROS in the islet β cell of the present application; Figure 9 The bar chart of the fluorescence quantitative analysis of the determination of the ROS level in the islet β cell of the present application; Figure 10 The fluorescence microscopic image of the determination of the mitochondrial membrane potential in the islet β cell of the present application; Figure 11 The bar chart of the fluorescence quantitative analysis of the determination of the mitochondrial membrane potential in the islet β cell of the present application; Figure 12 The bar chart of the determination of the insulin secretion content in the islet β cell of the present application; Figure 13 The bar chart of the determination of the glucose consumption in the islet β cell of the present application. DETAILED DESCRIPTION

[0016] The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, but not all the embodiments of the present application. Based on the embodiments of the present application, all the other embodiments obtained by those skilled in the art without creative work fall within the scope of the present application.

[0017] The following embodiments are prepared according to the processes of Figure 2 and Figure 3 The preparation principles of the following embodiments are as followsFigure 1 as shown.

[0018] Example 1 Preparation of fullerene nanomaterials with glycolipid metabolism regulation ability: Step S1, 0.2 g of fullerene was put into 30 mL of anhydrous N,N-dimethylformamide and ultrasonically shaken for 20 min to uniformly disperse the fullerene to obtain a fullerene dispersion liquid. Then 3.0 g of reaction raw material thiomalic acid and 2.7 g of initiator dicumyl peroxide were added to a three-necked flask, and the fullerene dispersion liquid was added under a nitrogen atmosphere. The temperature was raised to 140°C, and the reaction was kept at this temperature for 20 h. After the reaction was completed, the temperature was cooled to room temperature. The unreacted fullerene was removed by centrifugation at 6000 rpm for 10 min. The supernatant was dialyzed with deionized water and ethanol three times, respectively. Then the dialysis bag was spin-dried to obtain carboxylated fullerene. Step S2, 0.2 g of carboxylated fullerene and 5 mL of anhydrous N,N-dimethylformamide and 10 mL of thionyl chloride were added to a three-necked flask. After acyl chloride reaction at 40°C for 20 h, the reaction was completed. After standing for 12 h, a precipitate appeared. The supernatant was discarded by centrifugation at 8000 rpm for 10 min and washed with a small amount of tetrahydrofuran. The precipitate was dried at 50°C for 24 h to obtain acyl chloride fullerene. Step S3, 0.2 g of acyl chloride fullerene was mixed with 20 mL of anhydrous N,N-dimethylformamide and ultrasonically shaken for 20 min to uniformly disperse the fullerene to obtain an acyl chloride fullerene dispersion liquid. Then 3.0 g of freshly prepared free metformin, 3 mL of anhydrous N,N-dimethylformamide and 5 mL of triethylamine were added to a three-necked flask. After stirring and mixing at room temperature for 1 h, the acyl chloride fullerene dispersion liquid was added. The temperature was raised to 120°C, and the reaction was kept at this temperature for 72 h. After the reaction was completed, the temperature was cooled to room temperature. The supernatant was dialyzed with deionized water and ethanol three times, respectively. After drying at 40°C under reduced pressure for 24 h, fullerene nanomaterials with glycolipid metabolism regulation ability were obtained.

[0019] Example 2 Step S1, 0.2 g of fullerene was put into 30 mL of anhydrous N,N-dimethylformamide and ultrasonically shaken for 30 min to uniformly disperse the fullerene to obtain a fullerene dispersion liquid. Then 3.0 g of reaction raw material thiomalic acid and 2.7 g of initiator dicumyl peroxide were added to a three-necked flask, and the fullerene dispersion liquid was added under a nitrogen atmosphere. The temperature was raised to 140°C, and the reaction was kept at this temperature for 48 h. After the reaction was completed, the temperature was cooled to room temperature. The unreacted fullerene was removed by centrifugation at 8000 rpm for 15 min. The supernatant was dialyzed with deionized water and ethanol three times, respectively. Then the dialysis bag was spin-dried to obtain carboxylated fullerene. Step S2, 0.2 g carboxylated fullerene and 5 mL anhydrous N,N-dimethylformamide 10 mL thionyl chloride were added into a three-necked flask, and after acyl chloride reaction at 40 ℃ for 20 h, the reaction was completed, and after standing for 12 h, a precipitate appeared, which was centrifuged at 10,000 rpm for 15 min, the supernatant was discarded and washed with a small amount of tetrahydrofuran, and the precipitate was dried at 60 ℃ for 12 h to obtain acyl chloride fullerene; Step S3, 0.2 g acyl chloride fullerene was mixed with 20 mL anhydrous N,N-dimethylformamide, and ultrasonic oscillation was performed for 30 min to uniformly disperse the fullerene, to obtain an acyl chloride fullerene dispersion liquid, and then 3.0 g freshly prepared free metformin, 3 mL anhydrous N,N-dimethylformamide and 5 mL triethylamine were added into a three-necked flask, and after stirring and mixing at room temperature for 2 h, the acyl chloride fullerene dispersion liquid was added, and the temperature was raised to 120 ℃, and the reaction was carried out at this temperature for 72 h, and after the reaction was completed, the temperature was lowered to room temperature, and dialysis was performed with deionized water and ethanol three times respectively, and 50 ℃ under reduced pressure drying was carried out for 48 h to obtain a fullerene nanomaterial with sugar and lipid metabolism regulation ability.

[0020] The FT-IR spectra of (a) fullerene, (b) carboxylated fullerene, (c) metformin and (d) fullerene nanomaterial with sugar and lipid metabolism regulation ability in Example 1 were determined by using a Fourier transform infrared spectrometer, as shown in Figure 4 By comparing the infrared spectra of curve (a) and curve (b), it was found that C 60 -COOH appeared at 1656 cm -1 , which can be attributed to the C-O stretching vibration of carboxylic acid group, and the peak at 3436 cm -1 belongs to the stretching vibration of O-H, which proves that the carboxylic acid derivative is successfully synthesized. The absorption peak at 2923 cm -1 corresponds to the stretching vibration of C-H, which is -CH2- in the reactant, which indicates the successful preparation of fullerene carboxylic acid derivative. In curve (c), 3372 cm -1 , 3294 cm -1 and 3174 cm -1 respectively correspond to the stretching vibration of primary amine and secondary amine N-H, and there is a small split, which may be due to the Fermi resonance caused by the peak at 1624 cm -1 . The peak at 1624 cm -1 is the bending vibration of C-H. The absorption peaks at 1582 cm -1 and 1472 cm -1 can be attributed to the stretching vibration of two imine bonds C=N of the connected metformin. In curve (d), 1654 cm -1the absorption band in the region of 1543 cm -1 and 1443 cm -1 may be attributed to the stretching vibration of the imine bond C=N of the attached metformin. Comparing curve (c) and curve (d), it can be found that the position of the imine has a blue shift, which may be caused by the enhanced conjugation after the attachment of the carboxyl derivative. These results indicate that metformin is bonded to the surface of fullerene through amidation reaction.

[0021] The SEM images of (a) and (b) in Example 1 are fullerene, (c) and (d) are carboxylated fullerene, (e) and (f) are fullerene nanomaterials with the ability to regulate glycolipid metabolism, as shown in Figure 5 ; the synthesized fullerene derivatives significantly improve the agglomeration state of fullerene in water and change the basic morphology.

[0022] The fluorescence emission spectra of the carboxylated fullerene and the fullerene nanomaterials with the ability to regulate glycolipid metabolism prepared in Example 1 were measured in ethanol, as shown in Figure 6 , with an excitation wavelength of 365 nm; Figure 6 It can be seen from the above that the synthesized fullerene derivatives produce fluorescence, while the raw material fullerene and metformin do not produce fluorescence, and the fluorescence produced by the intermediate product is different from the spectrum of the final synthetic product, which can indirectly prove the successful synthesis of the material. The production of fluorescence indicates that the molecule forms a conjugate, i.e. the electron transfer and balance ability are enhanced, and the original fullerene plays a better role in free radical balance.

[0023] The toxicity of the fullerene nanomaterials with the ability to regulate glycolipid metabolism prepared in Example 1 on pancreatic beta cells (MIN6) was detected using CCK8 experiment, and the cells were treated with different concentrations of fullerene nanomaterials with the ability to regulate glycolipid metabolism, as shown in Figure 7 , the fullerene nanomaterials with the ability to regulate glycolipid metabolism had little effect on the activity of the cells, and when the concentration of the fullerene nanomaterials with the ability to regulate glycolipid metabolism was increased to 0.15 mg / mL, the cell survival rate was increased to 106.5%.

[0024] The mouse islet β cells induced by STZ (streptozotocin, 1 mM, 4 h) under oxidative stress, treatment group: islet β cells treated with 50 μg / mL of the fullerene nanomaterial with glycolipid metabolism regulation ability prepared in Example 1; model group: islet cells treated with 50 μg / mL of PBS in the STZ (1 mM, 4 h) induced oxidative stress model group, untreated normal group: healthy mouse islet β cells without STZ induction; metformin positive control group, and the fluorescence microscopic image of intracellular reactive oxygen species (ROS) level was determined by fluorescence microscopy, as shown in Figure 8 Compared with the model group, the intracellular ROS level of the cells treated with 50 ug / mL of the fullerene nanomaterial with glycolipid metabolism regulation ability was significantly reduced, proving that the fullerene nanomaterial with glycolipid metabolism regulation ability has a good relieving effect on the oxidative stress caused by STZ, while the positive control group metformin has no obvious effect on relieving oxidative stress.

[0025] GraphPad Prism 8 software was used for plotting, t-test and ANOVA were used for statistical significance analysis, p<0.05 was considered to have statistical significance, ** meant p<0.01 compared with the control group, # meant p<0.05 compared with the model group, and the fluorescence quantification of intracellular ROS level was determined, the treatment group: STZ (1 mM, 4 h) induced oxidative stress islet β cells treated with 50 μg / mL of the fullerene nanomaterial with glycolipid metabolism regulation ability prepared in Example 1; model group: STZ (1 mM, 4 h) induced oxidative stress model group, untreated normal group, metformin positive control group, and the results are shown in Figure 9 .

[0026] STZ (1 mM, 4 h) was used to induce mitochondrial damage, and the fullerene nanomaterial with glycolipid metabolism regulation ability prepared in Example 1 (50 μg / mL) and metformin (50 μg / mL) were used for treatment, and normal untreated group and STZ model group were set; the fluorescence microscopic image of the change of mitochondrial membrane potential of islet β cells was determined, as shown in Figure 10 JC-10 fluorescence probe in cells with normal mitochondrial function without damage, mitochondrial potential is maintained at high potential, JC-10 polymer produces red fluorescence in mitochondrial matrix, and once the cell mitochondria are damaged, the mitochondrial potential membrane potential decreases, the polymer forms green fluorescence in the mitochondrial matrix, and the relative proportion of red / green fluorescence can be used to judge the decrease of membrane potential.

[0027] GraphPad Prism 8 software was used for plotting, t-test and ANOVA were used for statistical significance analysis, ** means p < 0.01 compared with the control group, ### means p < 0.001 compared with the model group, $$ means p < 0.01 compared with the treatment group, STZ (1 mM, 4 h) was used to induce mitochondrial damage, and fullerene nanomaterials with glycolipid metabolism regulation ability prepared in Example 1 (50 μg / mL) and metformin (50 μg / mL) were used for treatment, and normal control group and STZ model group were set; the fluorescence quantification of the high and low mitochondrial membrane potential of islet β cells was determined; as shown in Figure 11 After the islet β cells were treated with 1 mM STZ for 4 h, it was found that the mitochondrial membrane potential was significantly reduced by 42.4%, and then the SZT treated cells were treated with 50 μg / ml fullerene nanomaterials with antioxidant activity and glycolipid metabolism regulation ability and 50 μg / ml metformin, respectively. It was found that fullerene nanomaterials with glycolipid metabolism regulation ability could restore the high mitochondrial potential, while metformin also increased the membrane potential but did not significantly change.

[0028] Islet β cells can secrete insulin and adjust the secretion amount when the blood glucose concentration changes, and the insulin secreted is the most important hormone for reducing blood glucose in the body. In this experiment, islet β cells (mouse islet β cell line) were used, which can secrete insulin under glucose stimulation, and the insulin content was quantitatively detected by ELISA (enzyme-linked immunosorbent assay). The experimental groups are as follows: normal control group (untreated cells), STZ model group (STZ induced (1 mM, 4 h) insulin secretion impaired cells), fullerene nanomaterials with glycolipid metabolism regulation ability prepared in Example 1 (10 μg / mL and 50 μg / mL), and metformin positive control group (50 μg / mL); the recovery of insulin secretion ability of damaged islet cells treated with drugs was determined, and the results are shown in Figure 12 The insulin concentration in the culture medium was reduced by 46.6% due to the induction of STZ, and then treated with fullerene nanomaterials with glycolipid metabolism regulation ability, when the concentration of fullerene nanomaterials with glycolipid metabolism regulation ability was 10 μg / mL and 50 μg / mL, the insulin concentration in the culture medium was further increased by 20.4% and 55.2% compared with the STZ model group. In this experiment, the common diabetes treatment drug metformin was selected as a positive control, and it was found that 50 μg / m metformin could increase the insulin concentration of the STZ induced group by 34.4%. Fullerene nanomaterials with glycolipid metabolism regulation ability improved insulin secretion ability much higher than metformin, which was 1.52 times the recovery ability of metformin under the same concentration conditions.

[0029] Glucose consumption is a common index for evaluating the ability of cells to utilize glucose. In the experiment, glucose oxidase method (GOD-POD) was used to reflect the uptake and metabolic ability of cells to glucose by detecting the reduction of glucose in the culture medium. The experimental groups were as follows: normal control group (untreated cells), STZ (1 mM, 4 h) model group, treatment group: STZ-induced damaged islet β cells treated with the fullerene nanomaterial with the ability to regulate glycolipid metabolism prepared in Example 1 (10 μg / mL and 50 μg / mL), negative control group of normal cells treated with the fullerene nanomaterial with the ability to regulate glycolipid metabolism prepared in Example 1 (10 μg / mL and 50 μg / mL), and metformin positive control group (50 μg / mL); the glucose consumption of cells was determined; the results are shown in Figure 13 As shown in Table 1, the glucose consumption level of the STZ model group was reduced by 39.41% compared with the control group, indicating that STZ would damage the cells and greatly reduce the ability of the cells to consume glucose. Compared with the STZ model group, the positive control group of 50 μg / ml metformin improved the glucose consumption level of islet β cells by 74.48%. The fullerene nanomaterial with the ability to regulate glycolipid metabolism slightly improved the glucose consumption ability of cells than metformin, and at the treatment concentration of 10 μg / ml and 50 μg / ml, it was improved by 70.84% and 89.55% respectively compared with the STZ-induced group. The islet β cells treated only with the fullerene nanomaterial with the ability to regulate glycolipid metabolism showed no significant change compared with the normal control group. It was indicated that the material had no significant effect on normal cells but showed good ability to restore the glucose consumption ability of cells induced by STZ.

[0030] In summary, the present application realizes the synergistic effect of the dual mechanisms of antioxidant and hypoglycemic by the chemical combination of fullerene and metformin, significantly improves the treatment effect, and has good safety and biocompatibility, and has important application value in the field of treating type II diabetes.

[0031] In the description of the specification, the description of the terms "one embodiment", "example", "specific example" and the like means that the specific features, structures, materials or characteristics described in connection with the embodiment or example are contained in at least one embodiment or example of the present application. In the specification, the illustrative description of the above terms does 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.

[0032] The above merely illustrates and describes the present application, and those skilled in the art can make various modifications or supplements to the described specific embodiments or adopt similar ways to replace, as long as the modifications or supplements do not deviate from the present application or exceed the scope defined by the present claims, and should belong to the protection scope of the present application.

Claims

1. A method for synthesizing fullerene nanomaterials with the ability to regulate glycolipid metabolism, characterized in that, Includes the following steps: Step S1: Fullerene is added to anhydrous N,N-dimethylformamide and ultrasonically vibrated to obtain a fullerene dispersion. Thiomalic acid and initiator dicumyl peroxide are then added to a flask. The fullerene dispersion is injected under a nitrogen atmosphere. The reaction is carried out at 140 °C for 20 h. After the reaction is completed, the carboxylated fullerene is obtained. Step S2: Add carboxylated fullerene and thionyl chloride to the flask and react at 40 °C for 24 h. After the reaction is complete, perform post-treatment to obtain acyl chloride fullerene. Step S3: Mix acyl chloride fullerene with anhydrous N,N-dimethylformamide and sonicate to obtain an acyl chloride fullerene dispersion. Then, add freshly prepared metformin, anhydrous N,N-dimethylformamide and triethylamine to a flask, stir and mix at room temperature, and then add the acyl chloride fullerene dispersion. React at 120 °C for 72 h. After the reaction is complete, perform post-processing to obtain fullerene nanomaterials with the ability to regulate glycolipid metabolism.

2. The method for synthesizing fullerene nanomaterials with glucose and lipid metabolism regulation capabilities according to claim 1, characterized in that, In step S1, the ratio of fullerene, anhydrous N,N-dimethylformamide, thiomalic acid, and dicumyl peroxide is 0.2 g:30 mL:3.0 g:2.7 g.

3. The method for synthesizing fullerene nanomaterials with glucose and lipid metabolism regulation capabilities according to claim 1, characterized in that, In step S2, the ratio of carboxylated fullerene, anhydrous N,N-dimethylformamide, and thionyl chloride is 0.2 g: 5 mL: 10 mL.

4. The method for synthesizing fullerene nanomaterials with glucose and lipid metabolism regulation capabilities according to claim 1, characterized in that, In step S3, the ratio of the amounts of acyl chloride fullerene, anhydrous N,N-dimethylformamide, freshly prepared metformin, and triethylamine is 0.2 g: 23 mL: 3.0 g: 5 mL.

5. The method for synthesizing fullerene nanomaterials with glucose and lipid metabolism regulation capabilities according to claim 1, characterized in that, The post-processing operation in step S1 is as follows: cool to room temperature, centrifuge at 6000-8000 rpm for 10-15 min to remove unreacted fullerenes, and dialyze the supernatant three times with deionized water and ethanol respectively, and then evaporate the suspension obtained in the dialysis bag to dryness.

6. The method for synthesizing fullerene nanomaterials with glucose and lipid metabolism regulation capabilities according to claim 1, characterized in that, The post-processing procedure in step S2 is as follows: let stand for 12 h to allow precipitation to occur, centrifuge at 8000-10000 rpm for 10-15 min, discard the supernatant and wash with a small amount of tetrahydrofuran, and dry the precipitate at 50-60 ℃ for 12-24 h.

7. The method for synthesizing fullerene nanomaterials with glucose and lipid metabolism regulation capabilities according to claim 1, characterized in that, The post-processing steps in step S3 are as follows: after cooling to room temperature, dialyze three times with deionized water and three times with ethanol, and then dry under reduced pressure at 40-50 °C for 24-48 h.

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