Method for high-throughput screening of xanthine oxidase inhibitor through magnetic ligand fishing and application of extract

Abstract

The invention discloses a method for high-throughput screening of a xanthine oxidase inhibitor by magnetic ligand fishing and application of an extract of the xanthine oxidase inhibitor, and the preparation method comprises the following steps: preparing amino-functionalized Fe3O4 (at) SiO2 nanoparticles from Fe3O4 nanoparticles, then preparing Fe3O4 (at) SiO2-XOD nanoparticles from the amino-functionalized Fe3O4 (at) SiO2 nanoparticles, mixing the Fe3O4 (at) SiO2-XOD nanoparticles with an extract solution of traditional Chinese medicinal materials, and co-incubating; washing and then incubating together with a positive drug allopurinol solution to obtain a specific effective component capable of being combined with xanthine oxidase in the traditional Chinese medicinal material extract; according to the magnetic ligand fishing high-throughput screening method of the xanthine oxidase inhibitor, effective components of the specific xanthine oxidase inhibitor in a medicine are accurately extracted, interference of other components is reduced, operation is easy, cost is low, the requirement for experimental instruments is low, and compared with other extraction methods, specificity is high, and the screening method is suitable for industrial production. Good application prospects are realized.

Description

Technical Field

[0001] The invention belongs to the technical field of screening active ingredients of traditional Chinese medicines, and relates to a method for high-throughput screening of xanthine oxidase inhibitors, and specifically relates to a method for high-throughput screening of xanthine oxidase inhibitors by magnetic ligand fishing and the application of extracts. Background Art

[0002] Uric acid (UA) in the body is divided into endogenous uric acid and exogenous uric acid. Endogenous uric acid is produced by the metabolism of nucleic acids and purine bodies in the body, accounting for about 80%-90% of the total uric acid metabolism in the body. Exogenous uric acid is ingested through foods such as beer, animal offal, and seafood, accounting for a smaller proportion of the total uric acid in the body. Under normal physiological and purine dietary conditions, the normal and stable operation of the uric acid metabolism system benefits from the dynamic balance of uric acid metabolism in the human body. When the purine metabolic pathway in the human body is unbalanced and uric acid is overproduced or excreted less, the serum uric acid level increases sharply and exists in the form of sodium salt, which eventually manifests as abnormal uric acid metabolism and evolves into hyperuricemia.

[0003] Clinically, gout is mainly prevented and treated by lowering the patient's uric acid level. Xanthine oxidase (XOD) is a key enzyme in the production of uric acid. Taking XOD inhibitors can significantly reduce uric acid levels to treat gout. Traditional XOD inhibitors such as allopurinol and febuxostat are first-line drugs for the treatment of gout. However, they often cause a series of side effects and may cause damage to the patient's liver and kidneys.

[0004] In addition to the XOD inhibitors that have been on the market, people have turned their research direction to traditional Chinese medicine and diet, and have made good progress. Natural products come from nature, and have the advantages of rich resources, easy access, and high safety. Studies have found that some natural products can achieve the purpose of lowering uric acid by inhibiting the activity of xanthine oxidase and regulating uric acid transporters. Screening out the components that inhibit XOD activity in these natural products will help their further development and utilization.

[0005] The existing technology usually uses an ultrasonic water extraction and then purification method to extract the target active ingredients from natural products. This extraction method will extract most of the active ingredients of traditional Chinese medicines, some of which do not have the effect of inhibiting xanthine oxidase, which is not conducive to studying the mechanism of the inhibitory effect of traditional Chinese medicines on xanthine oxidase; or it cannot completely screen out the effective ingredients in natural products, resulting in a waste of resources; at the same time, the existing extraction methods also have defects such as complex operation, low separation efficiency, and unsuitability for mass production. Summary of the invention

[0006] In view of the shortcomings of the prior art, the purpose of the present invention is to provide a method for high-throughput screening of xanthine oxidase inhibitors by magnetic ligand fishing and the application of extracts, which can accurately extract the effective ingredients of specific xanthine oxidase inhibitors in drugs, reduce the interference of other components, and have simple operation, high efficiency, high yield and low cost.

[0007] In order to achieve the above object, the present invention adopts the following technical solutions:

[0008] A method for high-throughput screening of xanthine oxidase inhibitors by magnetic ligand fishing comprises the following steps:

[0009] Step 1, selecting Chinese medicinal materials, extracting effective ingredients with hot water, collecting the filtrate, and then drying to obtain Chinese medicinal extract powder; the Chinese medicinal materials include any one of Eucommia ulmoides leaves, Pueraria root and Astragalus membranaceus;

[0010] Step 2: Preparation of amino-functionalized Fe 3 O 4 @SiO 2 Nanoparticles, by Fe 3 O 4 The molar ratio of glutaraldehyde to amino-functionalized Fe 3 O 4 @SiO 2 The nanoparticles were suspended in glutaraldehyde solution for 0.5-1 h to obtain a suspension, and then the xanthine oxidase and amino-functionalized Fe 3 O 4 @SiO 2 The mass ratio of nanoparticles was 1:(1-5) and xanthine oxidase was added. The mixture was stirred at room temperature for 30-50 min and XOD was fixed on the magnetic Fe by a typical glutaraldehyde activation procedure. 3 O 4 @SiO 2 On a solid carrier, magnetic separation, washing and drying to obtain Fe 3 O 4 @SiO 2 -XOD nanoparticles;

[0011] Step 3: Prepare a 1-10 mg / ml solution of the Chinese herbal extract powder, and 3 O 4 @SiO 2 The mass ratio of Fe-XOD nanoparticles is 1:(1-3) 3 O 4 @SiO 2 -XOD nanoparticles were suspended in the extract solution and incubated at 30-40°C for 0.5-3 h. 3O 4 @SiO 2 -XOD ligand complex, washed and dried for later use;

[0012] Step 4: Press Fe 3 O 4 @SiO 2 The mass ratio of -XOD ligand complex to allopurinol is (15-25):1. 3 O 4 @SiO 2 The -XOD ligand complex is suspended in a 0.1-1 mg / ml allopurinol solution, incubated at 30-40°C for 0.5-3 hours, subjected to magnetic separation, and the solution is collected and dried to obtain a xanthine oxidase inhibitor.

[0013] The present invention also has the following technical features:

[0014] Preferably, the hot water extraction in step 1 specifically includes soaking the Chinese medicinal materials in water at a mass ratio of 1:(5-10), standing for 0.5-1h, heating to 70-100°C, and keeping warm for 1-4h.

[0015] Preferably, the amino-functionalized Fe 3 O 4 @SiO 2 The method for preparing nanoparticles includes:

[0016] Take Fe 3 O 4 The nanoparticles were dispersed in an ethanol solution with a volume percentage of 75-80%. After being uniformly dispersed by ultrasonic, ammonia water was added to adjust the pH of the system to 8-10. 3 O 4 The mass ratio of nanoparticles to tetraethyl orthosilicate is 1:(1-4) tetraethyl orthosilicate is added, stirred at room temperature for 6-8 hours, and then adsorbed and recovered by magnet to obtain Fe 3 O 4 @SiO 2 The nanoparticles are washed and dried for later use;

[0017] The Fe 3 O 4 @SiO 2 The nanoparticles were dispersed in anhydrous ethanol and then 3 O 4 @SiO 2 The mass volume ratio of nanoparticles to 3-aminopropyltriethoxysilane was 1: (0.1-1) 3-aminopropyltriethoxysilane was added, stirred at room temperature for 6-10 hours, and then adsorbed and recovered by a magnet and washed and dried to obtain amino-functionalized Fe 3O 4 @SiO 2 Nanoparticles.

[0018] Furthermore, the washing is washing with deionized water for 3 to 5 times;

[0019] The drying is vacuum freeze drying.

[0020] Preferably, the mass volume concentration of the glutaraldehyde solution in step 2 is 25%;

[0021] Preferably, in step 2, xanthine oxidase is added to deionized water to prepare an aqueous solution with a concentration of 2.5 U / ml and then added to the suspension.

[0022] Preferably, the washing in step 2 is washing with deionized water for 3 to 5 times.

[0023] Preferably, the washing in step 3 is performed with 1×PBS for 3 to 5 times.

[0024] Preferably, the drying in step 1, step 2, step 3 and step 4 is vacuum freeze drying.

[0025] The present invention also protects the use of a xanthine oxidase inhibitor screened by the above method in food or medicine.

[0026] Compared with the prior art, the present invention has the following technical effects:

[0027] The screening method of xanthine oxidase inhibitors of the present invention uses magnetic ligand fishing technology as the main body to 3 O 4 Nanoparticles prepared from Fe 3 O 4 @SiO 2 -XOD nanoparticles are mixed with the extract solution of Chinese medicinal materials, incubated together, and then washed and incubated together with the positive drug allopurinol solution. With the help of the specific binding of xanthine oxidase, the obtained products are all effective ingredients that can specifically bind to xanthine oxidase. The specific xanthine oxidase inhibitor effective ingredients in the drug are accurately extracted, and the interference of other components of the extract on the mechanism of inhibiting xanthine oxidase in the study of Chinese medicinal materials can be excluded. It can provide certain theoretical and technical support for the study of drugs and means for the treatment of gout or hyperuricemia;

[0028] The screening method for xanthine oxidase inhibitors of the present invention is simple to operate, low in cost, can accurately and completely extract the specific xanthine oxidase inhibitor effective ingredients in drugs, has high extraction efficiency and yield, has low requirements on production instruments, and has excellent application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 The HPLC data of the whole water extract of Eucommia ulmoides leaves in Example 1;

[0030] Figure 2 This is a HPLC data diagram of the product obtained by the method of high-throughput screening of xanthine oxidase inhibitors in Eucommia ulmoides leaves using magnetic ligand fishing in Example 1;

[0031] Figure 3 The inhibitory effects of the products obtained in Examples 1 to 4 on xanthine oxidase. DETAILED DESCRIPTION

[0032] The specific contents of the present invention are further explained in detail below in conjunction with the examples. The reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art. The test methods in the following examples where specific experimental conditions are not specified are usually based on conventional experimental conditions. Unless otherwise specified, the reagents and raw materials used in the present invention can be obtained commercially.

[0033] Embodiment 1:

[0034] This embodiment provides a method for high-throughput screening of xanthine oxidase inhibitors by magnetic ligand fishing, comprising the following steps:

[0035] Step 1: Select dried Eucommia ulmoides leaves, crush them into powder, soak them in water at a mass ratio of 1:10, let them stand for 0.5 hours, heat them to 100°C, keep them warm for 4 hours, collect the filtrate, and then freeze-dry them to obtain a Chinese medicine extract powder;

[0036] The obtained product was subjected to high performance liquid chromatography, and the results were as follows: Figure 1 shown.

[0037] Step 2: Take Fe 3 O 4 The nanoparticles were dispersed in 75% ethanol solution by volume. After being dispersed uniformly by ultrasonic, ammonia water was added to adjust the pH of the system to 10. 3 O 4 The mass ratio of nanoparticles to tetraethyl orthosilicate was 1:1. Tetraethyl orthosilicate was added and stirred at room temperature for 6 h. Then, it was adsorbed and recovered by a magnet to obtain Fe 3 O 4 @SiO 2 The nanoparticles were washed three times with deionized water and freeze-dried in vacuum for later use;

[0038] The Fe 3 O 4 @SiO 2 The nanoparticles were dispersed in anhydrous ethanol and then 3 O4 @SiO 2 The nanoparticles and 3-aminopropyltriethoxysilane were added in a mass volume ratio of 1:1, stirred at room temperature for 6 h, and then adsorbed and recovered by a magnet and washed with deionized water three times, and then freeze-dried in a vacuum to obtain amino-functionalized Fe 3 O 4 @SiO 2 Nanoparticles;

[0039] Press Fe 3 O 4 The amino-functionalized Fe 3 O 4 @SiO 2 The nanoparticles were suspended in a 25% mass volume concentration glutaraldehyde solution for 0.5 h to obtain a suspension, and then the xanthine oxidase and amino-functionalized Fe 3 O 4 @SiO 2 The mass ratio of nanoparticles was 1:1, and a 2.5U / ml xanthine oxidase aqueous solution was added. The mixture was stirred at room temperature for 30 min, and XOD was fixed on the magnetic Fe by a typical glutaraldehyde activation procedure. 3 O 4 @SiO 2 The solid carrier was magnetically separated, washed with deionized water three times, and vacuum freeze-dried to obtain Fe 3 O 4 @SiO 2 -XOD nanoparticles;

[0040] Step 3: Prepare a 4.5 mg / ml solution of the Chinese herbal extract powder and Fe 3 O 4 @SiO 2 The mass ratio of Fe-XOD nanoparticles was 1:2. 3 O 4 @SiO 2 -XOD nanoparticles were suspended in the extract solution and incubated at 30 °C for 1 h. 3 O 4 @SiO 2 -XOD ligand complex was washed three times with 1× PBS and freeze-dried in vacuum for later use;

[0041] Step 4: Press Fe 3 O 4 @SiO 2 The mass ratio of -XOD ligand complex to allopurinol was 15:1. 3 O 4@SiO 2 The -XOD ligand complex was suspended in a 0.1 mg / ml allopurinol solution, incubated at 30°C for 0.5 h, subjected to magnetic separation, and the solution was collected and vacuum freeze-dried to obtain a xanthine oxidase inhibitor.

[0042] The obtained product was subjected to high performance liquid chromatography, and the results were as follows: Figure 2 As shown, Figure 1 In comparison, the number of peaks was significantly reduced, indicating that the effective components that could not specifically bind to xanthine oxidase were removed during the screening process.

[0043] The xanthine oxidase inhibitory activity was verified and the result was that the inhibition rate was 85.12%.

[0044] Embodiment 2:

[0045] This embodiment provides a method for high-throughput screening of xanthine oxidase inhibitors by magnetic ligand fishing, comprising the following steps:

[0046] Step 1: Select dried Pueraria root, crush it into powder, soak it in water at a mass ratio of 1:5, let it stand for 1 hour, heat it to 70°C, keep it warm for 2 hours, collect the filtrate, and then freeze-dry it to obtain a Chinese medicine extract powder;

[0047] Step 2: Take Fe 3 O 4 The nanoparticles were dispersed in an ethanol solution with a volume percentage of 80%. After being uniformly dispersed by ultrasonic, ammonia water was added to adjust the pH of the system to 8. 3 O 4 The mass ratio of nanoparticles to tetraethyl orthosilicate was 1:2. Tetraethyl orthosilicate was added and stirred at room temperature for 7 h. Then, it was adsorbed and recovered by a magnet to obtain Fe 3 O 4 @SiO 2 The nanoparticles were washed four times with deionized water and freeze-dried in vacuum for later use;

[0048] The Fe 3 O 4 @SiO 2 The nanoparticles were dispersed in anhydrous ethanol and then 3 O 4 @SiO 2 The nanoparticles and 3-aminopropyltriethoxysilane were added in a mass volume ratio of 1:0.1, stirred at room temperature for 8 h, and then adsorbed and recovered by a magnet and washed with deionized water for 4 times, and then freeze-dried in vacuum to obtain amino-functionalized Fe 3 O 4 @SiO 2 Nanoparticles;

[0049] Press Fe 3 O 4 The amino-functionalized Fe 3 O 4 @SiO 2 The nanoparticles were suspended in a 25% mass volume concentration glutaraldehyde solution for 0.8 h to obtain a suspension, and then the xanthine oxidase and amino-functionalized Fe 3 O 4 @SiO 2 The mass ratio of nanoparticles was 1:3, and a 2.5U / ml xanthine oxidase aqueous solution was added. The mixture was stirred at room temperature for 40 min, and XOD was fixed on the magnetic Fe by a typical glutaraldehyde activation procedure. 3 O 4 @SiO 2 The solid carrier was magnetically separated, washed with deionized water 4 times, and vacuum freeze-dried to obtain Fe 3 O 4 @SiO 2 -XOD nanoparticles;

[0050] Step 3: Prepare a 1 mg / ml solution of the Chinese herbal extract powder and Fe 3 O 4 @SiO 2 The mass ratio of Fe-XOD nanoparticles was 1:1. 3 O 4 @SiO 2 -XOD nanoparticles were suspended in the extract solution and incubated at 40 °C for 0.5 h. 3 O 4 @SiO 2 -XOD ligand complex was washed four times with 1× PBS and freeze-dried in vacuum for later use;

[0051] Step 4: Press Fe 3 O 4 @SiO 2 The mass ratio of the -XOD ligand complex to allopurinol was 25:1. 3 O 4 @SiO 2 The -XOD ligand complex was suspended in a 1 mg / ml allopurinol solution, incubated at 40°C for 3 h, subjected to magnetic separation, and the solution was collected and vacuum freeze-dried to obtain a xanthine oxidase inhibitor.

[0052] The xanthine oxidase inhibitory activity of the product was verified, and the result obtained was that the inhibition rate was 79.63%.

[0053] Embodiment 3:

[0054] This embodiment provides a method for high-throughput screening of xanthine oxidase inhibitors by magnetic ligand fishing, comprising the following steps:

[0055] Step 1: Select dried Astragalus, crush it into powder, soak it in water at a mass ratio of 1:8, let it stand for 0.8 hours, heat it to 80°C, keep it warm for 1 hour, collect the filtrate, and then freeze-dry it to obtain a Chinese medicine extract powder;

[0056] Step 2: Take Fe 3 O 4 The nanoparticles were dispersed in 78% ethanol solution by volume. After being dispersed uniformly by ultrasonic, ammonia water was added to adjust the pH of the system to 9. 3 O 4 The mass ratio of nanoparticles to tetraethyl orthosilicate was 1:4. Tetraethyl orthosilicate was added and stirred at room temperature for 8 h. Then, it was adsorbed and recovered by a magnet to obtain Fe 3 O 4 @SiO 2 The nanoparticles were washed five times with deionized water and freeze-dried in vacuum for later use;

[0057] The Fe 3 O 4 @SiO 2 The nanoparticles were dispersed in anhydrous ethanol and then 3 O 4 @SiO 2 The nanoparticles and 3-aminopropyltriethoxysilane were added in a mass volume ratio of 1:0.5, stirred at room temperature for 10 h, and then adsorbed and recovered by a magnet and washed with deionized water for 5 times, and then freeze-dried in vacuum to obtain amino-functionalized Fe 3 O 4 @SiO 2 Nanoparticles;

[0058] Press Fe 3 O 4 The amino-functionalized Fe 3 O 4 @SiO 2 The nanoparticles were suspended in a 25% mass volume concentration of glutaraldehyde solution for 1 h to obtain a suspension, and then the xanthine oxidase and amino-functionalized Fe 3 O 4 @SiO 2The mass ratio of nanoparticles was 1:5, and a 2.5U / ml xanthine oxidase aqueous solution was added. The mixture was stirred at room temperature for 50 min, and XOD was fixed on the magnetic Fe by a typical glutaraldehyde activation procedure. 3 O 4 @SiO 2 The solid carrier was magnetically separated, washed with deionized water 5 times, and vacuum freeze-dried to obtain Fe 3 O 4 @SiO 2 -XOD nanoparticles;

[0059] Step 3: Prepare a 10 mg / ml solution of the Chinese herbal extract powder and Fe 3 O 4 @SiO 2 The mass ratio of Fe-XOD nanoparticles was 1:3. 3 O 4 @SiO 2 -XOD nanoparticles were suspended in the extract solution and incubated at 35 °C for 2 h. 3 O 4 @SiO 2 -XOD ligand complex was washed five times with 1× PBS and freeze-dried in vacuum for later use;

[0060] Step 4: Press Fe 3 O 4 @SiO 2 The mass ratio of -XOD ligand complex to allopurinol was 20:1. 3 O 4 @SiO 2 The -XOD ligand complex was suspended in a 0.5 mg / ml allopurinol solution, incubated at 35°C for 2 h, subjected to magnetic separation, and the solution was collected and vacuum freeze-dried to obtain a xanthine oxidase inhibitor.

[0061] The xanthine oxidase inhibitory activity of the product was verified, and the result obtained was that the inhibition rate was 59.87%.

[0062] Embodiment 4:

[0063] This embodiment provides a method for high-throughput screening of xanthine oxidase inhibitors by magnetic ligand fishing, comprising the following steps:

[0064] Step 1: Select dried Eucommia ulmoides leaves, crush them into powder, soak them in water at a mass ratio of 1:6, let them stand for 0.6 hours, heat them to 90°C, keep them warm for 3 hours, collect the filtrate, and then freeze-dry them to obtain a Chinese medicine extract powder;

[0065] Step 2: Take Fe 3 O4 The nanoparticles were dispersed in a 75% ethanol solution by volume. After being evenly dispersed by ultrasonic, ammonia water was added to adjust the pH of the system to 8. 3 O 4 The mass ratio of nanoparticles to tetraethyl orthosilicate was 1:3. Tetraethyl orthosilicate was added and stirred at room temperature for 8 h. Then, it was adsorbed and recovered by a magnet to obtain Fe 3 O 4 @SiO 2 The nanoparticles were washed five times with deionized water and freeze-dried in vacuum for later use;

[0066] The Fe 3 O 4 @SiO 2 The nanoparticles were dispersed in anhydrous ethanol and then 3 O 4 @SiO 2 The nanoparticles and 3-aminopropyltriethoxysilane were added in a mass volume ratio of 1:0.3, stirred at room temperature for 9 h, and then adsorbed and recovered by a magnet and washed with deionized water for 5 times, and then freeze-dried in vacuum to obtain amino-functionalized Fe 3 O 4 @SiO 2 Nanoparticles;

[0067] Press Fe 3 O 4 The amino-functionalized Fe 3 O 4 @SiO 2 The nanoparticles were suspended in a 25% mass volume concentration of glutaraldehyde solution for 1 h to obtain a suspension, and then the xanthine oxidase and amino-functionalized Fe 3 O 4 @SiO 2 The mass ratio of nanoparticles was 1:2, and a 2.5U / ml xanthine oxidase aqueous solution was added. The mixture was stirred at room temperature for 50 min, and XOD was fixed on the magnetic Fe by a typical glutaraldehyde activation procedure. 3 O 4 @SiO 2 The solid carrier was magnetically separated, washed with deionized water 5 times, and vacuum freeze-dried to obtain Fe 3 O 4 @SiO 2 -XOD nanoparticles;

[0068] Step 3: Prepare a 5 mg / ml solution of the Chinese herbal extract powder and Fe 3 O 4 @SiO2 The mass ratio of Fe-XOD nanoparticles was 1:1. 3 O 4 @SiO 2 -XOD nanoparticles were suspended in the extract solution and incubated at 40 °C for 0.5 h. 3 O 4 @SiO 2 -XOD ligand complex was washed five times with 1× PBS and freeze-dried in vacuum for later use;

[0069] Step 4: Press Fe 3 O 4 @SiO 2 The mass ratio of the -XOD ligand complex to allopurinol was 25:1. 3 O 4 @SiO 2 The -XOD ligand complex was suspended in a 0.8 mg / ml allopurinol solution, incubated at 40°C for 0.5 h, subjected to magnetic separation, and the solution was collected and vacuum freeze-dried to obtain a xanthine oxidase inhibitor.

[0070] The xanthine oxidase inhibitory activity of the product was verified, and the result obtained was that the inhibition rate was 81.32%.

[0071] It should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it; without departing from the concept of the present invention, the deduction or replacement made by those skilled in the art shall fall within the protection scope of the present invention.

Claims

1. A method for high-throughput screening of xanthine oxidase inhibitors by magnetic ligand fishing, characterized in that: The following steps are involved: Step 1, selecting Chinese medicinal materials, extracting effective ingredients with hot water, collecting the filtrate, and then drying to obtain Chinese medicinal extract powder; the Chinese medicinal materials include any one of Eucommia ulmoides leaves, Pueraria root and Astragalus membranaceus; Step 2, preparing amino-functionalized Fe3O4@SiO2 nanoparticles, suspending the amino-functionalized Fe3O4@SiO2 nanoparticles in a glutaraldehyde solution at a molar ratio of Fe3O4 to glutaraldehyde of 1:(1-2) for 0.5-1h to obtain a suspension, then adding xanthine oxidase at a mass ratio of xanthine oxidase to amino-functionalized Fe3O4@SiO2 nanoparticles of 1:(1-5), stirring at room temperature for 30-50min, fixing XOD on a magnetic Fe3O4@SiO2 solid carrier through a typical glutaraldehyde activation procedure, magnetically separating, washing and drying to obtain Fe3O4@SiO2-XOD nanoparticles; Step 3: Prepare a 1-10 mg / ml solution of the Chinese herbal medicine extract powder, suspend the Fe3O4@SiO2-XOD nanoparticles in the extract solution according to the mass ratio of the Chinese herbal medicine extract powder to the Fe3O4@SiO2-XOD nanoparticles of 1:(1-3), incubate at 30-40°C with shaking for 0.5-3h, magnetically separate the Fe3O4@SiO2-XOD ligand complex, wash and dry it for later use; Step 4: According to the mass ratio of Fe3O4@SiO2-XOD ligand complex to allopurinol of (15-25):1, the Fe3O4@SiO2-XOD ligand complex is suspended in 0.1-1 mg / ml allopurinol solution, incubated at 30-40°C for 0.5-3h, subjected to magnetic separation, and the solution is collected and dried to obtain a xanthine oxidase inhibitor.

2. The method for high-throughput screening of xanthine oxidase inhibitors by magnetic ligand fishing as claimed in claim 1, characterized in that: The hot water extraction described in step 1 specifically includes soaking the Chinese medicinal materials in water at a mass ratio of 1:(5-10), leaving it to stand for 0.5-1h, heating it to 70-100°C, and keeping it warm for 1-4h.

3. The method for high-throughput screening of xanthine oxidase inhibitors by magnetic ligand fishing as claimed in claim 1, characterized in that: The preparation method of amino-functionalized Fe3O4@SiO2 nanoparticles described in step 2 comprises: Fe3O4 nanoparticles are dispersed in an ethanol solution with a volume percentage of 75-80%, and after being uniformly dispersed by ultrasonication, ammonia water is added to adjust the pH value of the system to 8-10, and then tetraethyl orthosilicate is added according to a mass ratio of Fe3O4 nanoparticles to tetraethyl orthosilicate of 1:(1-4), and after stirring at room temperature for 6-8 hours, a magnet is used for adsorption recovery to obtain Fe3O4@SiO2 nanoparticles, which are washed and dried for later use; The Fe3O4@SiO2 nanoparticles prepared above were dispersed in anhydrous ethanol, and then 3-aminopropyltriethoxysilane was added at a mass volume ratio of Fe3O4@SiO2 nanoparticles to 3-aminopropyltriethoxysilane of 1:(0.1~1). After stirring at room temperature for 6~10 hours, the mixture was adsorbed and recovered by a magnet and washed and dried to obtain amino-functionalized Fe3O4@SiO2 nanoparticles.

4. The method for high-throughput screening of xanthine oxidase inhibitors by magnetic ligand fishing as claimed in claim 3, characterized in that: The washing is washing with deionized water for 3 to 5 times; The drying is vacuum freeze drying.

5. The method for high-throughput screening of xanthine oxidase inhibitors by magnetic ligand fishing as claimed in claim 1, characterized in that: The mass volume concentration of the glutaraldehyde solution described in step 2 is 25%.

6. The method for high-throughput screening of xanthine oxidase inhibitors by magnetic ligand fishing as claimed in claim 1, characterized in that: In the step 2, xanthine oxidase is added to deionized water to prepare an aqueous solution with a concentration of 2.5 U / ml, and then added to the suspension.

7. The method for high-throughput screening of xanthine oxidase inhibitors by magnetic ligand fishing as claimed in claim 1, characterized in that: The washing in step 2 is washing with deionized water for 3 to 5 times.

8. The method for high-throughput screening of xanthine oxidase inhibitors by magnetic ligand fishing as claimed in claim 1, characterized in that: The washing described in step 3 was performed 3 to 5 times with 1×PBS.

9. The method for high-throughput screening of xanthine oxidase inhibitors by magnetic ligand fishing as claimed in claim 1, characterized in that: The drying described in step 1, step 2, step 3 and step 4 is vacuum freeze drying.

10. Use of a xanthine oxidase inhibitor screened by the method according to any one of claims 1 to 9 in functional foods or medicines.

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