A dual-functional particle-filled microgel and its preparation method and application

Abstract

The present invention discloses a bifunctional particle-filled microgel and its preparation method and application. The preparation method comprises: providing quinoa starch-pomegranate polyphenol nanoparticles; mixing inulin, water, 1-ethyl-(3-dimethylaminopropyl)carbodiimide, N-hydroxysuccinimide, and L-cysteine ​​to perform a cross-linking reaction to obtain thiolated inulin; mixing the quinoa starch-pomegranate polyphenol nanoparticles with the thiolated inulin, performing a self-cross-linking reaction, and obtaining a bifunctional particle-filled microgel. The particle-filled microgel provided by the present invention has colon targeting and glutathione stimulation responsiveness, and has a high resistant starch content, can be used to alleviate colitis and diabetes, and provides a theoretical basis for tailored intervention and precision nutrition for specific populations.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical materials, and in particular relates to a dual-functional particle-filled microgel and a preparation method and application thereof. Background Art

[0002] Ulcerative colitis (UC) is a typical inflammatory bowel disease characterized by immune relapse and gastrointestinal inflammation, accompanied by clinical symptoms such as diarrhea, rectal bleeding, and fatigue. These symptoms are linked to a pathogenesis that involves genetic susceptibility, epithelial barrier dysfunction, and intestinal dysbiosis. Patients often require drug enemas to relieve symptoms. However, in practice, drug enemas often face several challenges. The most important of these is that the drug may be excreted shortly after enema, thereby affecting its intestinal retention time, further compromising its absorption and efficacy. Nanoparticles have become a widely used tool for enhancing the delivery of poorly water-soluble functional ingredients. In particular, starch-based nanoparticle delivery systems have been demonstrated to be an effective strategy for improving the oral bioavailability of these ingredients.

[0003] In addition, starch is one of the essential macronutrients in the human diet and is also the main source of energy. However, the intake of rapidly digested starch can lead to a rapid increase in blood sugar after a meal, which has been shown to be an important cause of diet-related chronic diseases. Previous studies have shown that the intake of resistant starch (RS) does not cause a rapid increase in blood sugar after a meal. Therefore, the development of RS in starch staples can become a key solution to the problem of diet-related chronic diseases. Natural resistant starch RS1 is composed of starch wrapped in a cell wall constructed by crude fiber, which can effectively block amylase infiltration and reduce amylase hydrolysis of starch, thereby increasing RS content. Based on this, RS content can be increased by preparing bionic RS1-type starch.

[0004] Therefore, how to optimize existing starch-based nanoparticles so that they can be used to alleviate colitis and diabetes has become a direction that industry researchers have been working towards for a long time. Summary of the Invention

[0005] The main purpose of the present invention is to provide a dual-functional particle-filled microgel and its preparation method and application, so as to overcome the deficiencies in the prior art.

[0006] To achieve the aforementioned object of the invention, the technical solutions adopted by the present invention include:

[0007] One aspect of the present invention provides a method for preparing a bifunctional particle-filled microgel, comprising:

[0008] Provide quinoa starch-pomegranate polyphenol nanoparticles;

[0009] Mixing inulin, water, 1-ethyl-(3-dimethylaminopropyl)carbodiimide, N-hydroxysuccinimide, and L-cysteine ​​to undergo a cross-linking reaction to obtain thiolated inulin;

[0010] The quinoa starch-pomegranate polyphenol nanoparticles are mixed with thiolated inulin and subjected to an oxidation reaction to obtain a bifunctional particle-filled microgel.

[0011] Another aspect of the present invention provides a bifunctional particle-filled microgel prepared by the aforementioned preparation method.

[0012] Another aspect of the present invention provides the use of the aforementioned bifunctional particle-filled microgel in the preparation of a drug capable of treating colitis or diabetes.

[0013] Another aspect of the present invention provides a drug capable of treating colitis or diabetes, comprising the aforementioned bifunctional particle-filled microgel.

[0014] Compared with the prior art, the technical solution of the present invention has at least the following advantages:

[0015] The present invention uses thiolated inulin microgels to encapsulate quinoa starch-pomegranate polyphenol nanoparticles to form particle-filled microgels. The quinoa starch-pomegranate polyphenol nanoparticles prepared by magnetic field treatment have a high resistant starch content, can not only treat colitis but also have the effect of lowering blood sugar; the thiolated inulin prepared by a cross-linking reaction mediated by EDC and NHS has colon-targeting performance and glutathione stimulation responsiveness, is almost not released in the small intestine, and releases nanoparticles in the colon, thereby effectively alleviating colitis; and the present invention forms the thiolated inulin microgel by self-crosslinking, which is a simple method, has a simple gelation mechanism and in situ gelation performance, and compared with other chemically cross-linked microgels, the microgel of the present invention has fewer by-products and higher safety. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments recorded in the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0017] Figure 1a This is a comparison of the effects of the samples prepared in Example 1 of the present invention and Comparative Examples 1-3 on the colon length of mice;

[0018] Figure 1b This is a comparison chart of the effects of the samples prepared in Example 1 of the present invention and Comparative Examples 1-3 on the weight-to-length ratio of the mouse colon;

[0019] Figure 2a This is a comparison of the effects of the samples prepared in Example 1 of the present invention and Comparative Examples 1-3 on the secretion of inflammatory cytokine TNF-α in colitis mice;

[0020] Figure 2b This is a comparison of the effects of the samples prepared in Example 1 of the present invention and Comparative Examples 1-3 on the secretion of the inflammatory cytokine IL-1β in colitis mice;

[0021] Figure 2c This is a comparison of the effects of the samples prepared in Example 1 of the present invention and Comparative Examples 1-3 on the secretion of the inflammatory cytokine IL-6 in colitis mice;

[0022] Figure 2d This is a comparison of the effects of the samples prepared in Example 1 of the present invention and Comparative Examples 1-3 on the secretion of the inflammatory cytokine IL-10 in colitis mice;

[0023] Figure 3 This is a comparison chart of the effects of the samples prepared in Example 1 of the present invention and Comparative Examples 1-3 on the area under the blood glucose curve (AUC) level of diabetic mice. DETAILED DESCRIPTION

[0024] In view of the above problems existing in the prior art, the inventors of this case have conducted extensive and in-depth research and have provided a dual-functional particle-filled microgel and a preparation method thereof.

[0025] The inventors of this case discovered that microgels are deformable, soft, porous, three-dimensional microparticles with a stable structure of cross-linked biopolymer molecules, resulting from covalent bonds and strong non-covalent interactions. Microgels exhibit reversible swelling properties that respond to environmental changes, offering unique opportunities for oral delivery. Therefore, designing a novel delivery system that combines these advantages is expected to offer the best of both worlds: nanoparticles and microgels.

[0026] Furthermore, the inventors of this case also found that starch-based particle-filled microgels can encapsulate starch nanoparticles in a microgel matrix, which can be used as a bionic RS1-type starch to increase the resistant starch content and alleviate diabetes.

[0027] Based on this, the inventors of this case designed a dual-functional particle-filled microgel that can be used to relieve colitis and diabetes, and provide a theoretical basis for tailored intervention and precise nutrition for specific populations.

[0028] The following further explains the technical solution, its implementation process, and principles. However, it should be understood that the specific embodiments disclosed below are merely exemplary of the present invention, and that the present invention may be embodied in various forms. Therefore, the specific functional details disclosed herein should not be construed as limiting, but rather as a basis for the claims and a representative basis for teaching those skilled in the art to employ the present invention in various ways in virtually any appropriately detailed embodiment.

[0029] As one aspect of the technical solution of the present invention, a method for preparing a bifunctional particle-filled microgel includes:

[0030] Provide quinoa starch-pomegranate polyphenol nanoparticles;

[0031] Inulin, water, 1-ethyl-(3-dimethylaminopropyl)carbodiimide (EDC), N-hydroxysuccinimide (NHS), and L-cysteine ​​are mixed to undergo a cross-linking reaction to obtain thiolated inulin;

[0032] The quinoa starch-pomegranate polyphenol nanoparticles are mixed with thiolated inulin and subjected to an oxidation reaction to obtain a bifunctional particle-filled microgel.

[0033] It should be noted that EDC is used to activate inulin molecules, NHS is used to amidate inulin molecules, and the role of L-cysteine ​​is to thiolate inulin so that the inulin molecules carry thiol groups, so that the free sulfhydryl groups in the thiolated inulin can produce disulfide bonds through oxidation reaction, thereby completing the self-crosslinking reaction to obtain gel.

[0034] The present invention adopts EDC and NHS-mediated cross-linking reaction to prepare thiolated inulin. The thiolated inulin not only has colon targeting, but also has glutathione stimulation responsiveness due to the presence of disulfide bonds.

[0035] In some embodiments, the method for preparing quinoa starch-pomegranate polyphenol nanoparticles includes mixing pomegranate polyphenol with a quinoa starch solution, subjecting the mixture to a magnetic field to obtain a quinoa starch-pomegranate polyphenol complex solution, and then subjecting the mixture to ultrasonic treatment to obtain quinoa starch-pomegranate polyphenol nanoparticles. The quinoa starch-pomegranate polyphenol nanoparticles prepared by magnetic field treatment have a high resistant starch content and can not only treat colitis but also have the effect of lowering blood sugar.

[0036] In some preferred embodiments, the quinoa starch solution comprises quinoa starch and water, and the concentration of quinoa starch in the quinoa starch solution is 2.5-5 w / v%.

[0037] In some preferred embodiments, the mass ratio of the pomegranate polyphenols to quinoa starch is 1:5-1:20.

[0038] In some preferred embodiments, the magnetic field intensity of the magnetic field treatment is 5-20 T. If the magnetic field intensity is too strong, the ordered structure of the formed quinoa starch-pomegranate polyphenol complex will be reduced.

[0039] In some preferred embodiments, the magnetic field treatment time is 1-5 hours.

[0040] In some preferred embodiments, the ultrasonic power of the ultrasonic treatment is 100-500 W. Too strong ultrasonic power will destroy the structure of starch and increase digestibility.

[0041] In some preferred embodiments, the ultrasonic treatment time is 10-60 min.

[0042] In some embodiments, the preparation method of the quinoa starch-pomegranate polyphenol nanoparticles includes: adding pomegranate polyphenol to a quinoa starch solution and stirring to mix evenly, then placing in a magnetic field culture device for reaction, and after the reaction is completed, obtaining a quinoa starch-pomegranate polyphenol complex solution.

[0043] In some embodiments, the preparation method comprises: adjusting the pH value of the quinoa starch-pomegranate polyphenol complex solution to 6-9, and then performing ultrasonic treatment to obtain the quinoa starch-pomegranate polyphenol nanoparticles.

[0044] In some embodiments, the preparation method comprises: uniformly mixing an inulin aqueous solution, 1-ethyl-(3-dimethylaminopropyl)carbodiimide, and N-hydroxysuccinimide, and then adding L-cysteine ​​to perform a cross-linking reaction to obtain the thiolated inulin.

[0045] In some preferred embodiments, the concentration of the inulin aqueous solution is 1-5 w / v%.

[0046] In some preferred embodiments, the added amount of 1-ethyl-(3-dimethylaminopropyl)carbodiimide is 0.1-1 w / w% (based on the mass of inulin).

[0047] In some preferred embodiments, the added amount of N-hydroxysuccinimide is 0.1-1 w / w% (based on the mass of inulin).

[0048] In some preferred embodiments, the mass ratio of inulin to L-cysteine ​​is 20:1-2:1.

[0049] In some embodiments, the preparation method specifically includes: dissolving inulin in water, then adding EDC and NHS to obtain a mixture, stirring the mixture at room temperature for 15-30 minutes, then adding L-cysteine, adjusting the pH value to 5.0-6.0, and stirring at 200-500 rpm at room temperature for 1-3 hours to complete the cross-linking reaction.

[0050] In some embodiments, the preparation method includes: adding the quinoa starch-pomegranate polyphenol nanoparticles to a thiolated inulin solution, and then exposing the solution to air at room temperature for 1-3 hours (during this process, the free thiol groups in the thiolated inulin produce disulfide bonds through oxidation reaction, thereby completing the self-crosslinking reaction to obtain a gel), completing the self-crosslinking of the free thiol groups, and obtaining the bifunctional particle-filled microgel.

[0051] In some preferred embodiments, the mass ratio of the quinoa starch-pomegranate polyphenol nanoparticles to the thiolated inulin is 1:2-1:10.

[0052] In some preferred embodiments, the thiolated inulin solution is obtained by mixing the thiolated inulin with phosphate buffered saline (PBS).

[0053] In some more preferred embodiments, the concentration of the thiolated inulin solution is 2.5-5 w / v%.

[0054] In some more preferred embodiments, the pH value of the phosphate buffer is 7-8.

[0055] In some more specific embodiments, the preparation method comprises:

[0056] S1: adding pomegranate polyphenols to a quinoa starch solution and stirring evenly, placing the mixture in a magnetic field incubator and reacting with stirring for a specific time to obtain a quinoa starch-pomegranate polyphenol complex solution, then adjusting the pH value of the quinoa starch-pomegranate polyphenol complex solution to 6-9, and performing ultrasonic treatment using a probe ultrasonicator to obtain quinoa starch-pomegranate polyphenol nanoparticles;

[0057] S2: Dissolve inulin in sterile water, add appropriate amounts of EDC and NHS for activation and amidation of the inulin, and stir the mixture at room temperature for 15-30 minutes. Then, add L-cysteine, adjust the pH of the mixture to 5.0-6.0, and stir at room temperature at 200-500 rpm for 1-3 hours to complete the cross-linking reaction between L-cysteine ​​and inulin to form thiolated inulin;

[0058] S3: Dissolve the thiolated inulin in PBS (pH = 8) to form a thiolated inulin solution, add quinoa starch-pomegranate polyphenol nanoparticles to the thiolated inulin solution, and expose the mixture to air at room temperature for 1-3 hours to complete the self-crosslinking of the free thiol groups to obtain bifunctional particle-filled microgels.

[0059] As another aspect of the technical solution of the present invention, it also relates to a bifunctional particle-filled microgel prepared by the aforementioned preparation method.

[0060] As another aspect of the technical solution of the present invention, it also involves the use of the aforementioned bifunctional particle-filled microgel in the preparation of a drug having the function of treating colitis or diabetes.

[0061] As another aspect of the technical solution of the present invention, it also relates to a drug having the function of treating colitis or diabetes, which comprises the aforementioned bifunctional particle-filled microgel.

[0062] In summary, the present invention prepares a quinoa starch-pomegranate polyphenol complex through magnetic field treatment, further preparing quinoa starch-pomegranate polyphenol nanoparticles through ultrasonic induction. L-cysteine ​​is then dispersed in an inulin solution, forming thiolated inulin through a cross-linking reaction mediated by EDC and NHS. Finally, the quinoa starch-pomegranate polyphenol nanoparticles are dispersed in the thiolated inulin solution, and a self-crosslinked particle-filled microgel is formed through oxidation of free thiols. This invention, resulting in a dual-functional particle-filled microgel for relieving colitis and diabetes, provides a new and effective method for alleviating colitis and diabetes.

[0063] The present invention is further illustrated by way of examples below, but the invention is not limited to the scope of the examples. The reagents and raw materials used in the following examples are commercially available, and the experimental methods where specific conditions are not specified are generally carried out under conventional conditions or according to the conditions recommended by the respective manufacturers.

[0064] Example 1

[0065] (1) Preparation of quinoa starch-pomegranate polyphenol nanoparticles

[0066] A 5 w / v quinoa starch solution was prepared by adding 5 g of quinoa starch to 100 mL of ultrapure water. One g of pomegranate polyphenols was added to the solution and placed in an incubator with a magnetic field strength of 15 T and stirred for 5 hours to produce a quinoa starch-pomegranate polyphenol complex solution. The quinoa starch-pomegranate polyphenol complex solution was then adjusted to a pH of 7.5, and ultrasonic waves were applied at 500 W for 50 minutes to produce quinoa starch-pomegranate polyphenol nanoparticles.

[0067] (2) Preparation of thiolated inulin

[0068] 2.5 g of inulin was dissolved in 100 mL of sterile water. 0.1% (based on the mass of the inulin) of EDC and 0.1% (based on the mass of the inulin) of NHS were then added for activation and amidation of the inulin. The mixture was stirred at room temperature for 15 minutes. 1.0 g of L-cysteine ​​was added, the pH was adjusted to 5.0, and the mixture was stirred at 300 rpm for 3 hours to complete the cross-linking reaction between L-cysteine ​​and inulin to form thiolated inulin.

[0069] (3) Preparation of particle-filled microgels

[0070] 2.5g of thiolated inulin was dissolved in 100mL of PBS (pH 8) to form a thiolated inulin solution. 0.45g of quinoa starch-pomegranate polyphenol nanoparticles were dispersed in the thiolated inulin solution. The solution was exposed to air at room temperature for 1 hour to complete the self-crosslinking of the free thiol groups to form a particle-filled microgel.

[0071] Example 2

[0072] (1) Preparation of quinoa starch-pomegranate polyphenol nanoparticles

[0073] A 5 w / v quinoa starch solution was prepared by adding 5 g of quinoa starch to 100 mL of ultrapure water. 0.25 g of pomegranate polyphenols was added to the solution, which was then placed in an incubator with a magnetic field strength of 5 T and stirred for 1 hour to produce a quinoa starch-pomegranate polyphenol complex solution. The quinoa starch-pomegranate polyphenol complex solution was then adjusted to a pH of 7.5, and ultrasonic waves were applied at 100 W for 10 minutes to produce quinoa starch-pomegranate polyphenol nanoparticles.

[0074] (2) Preparation of thiolated inulin

[0075] 2.5 g of inulin was dissolved in 100 mL of sterile water. 0.1% (based on the weight of the inulin) of EDC and 0.1% (based on the weight of the inulin) of NHS were then added for activation and amidation of the inulin. The mixture was stirred at room temperature for 15 minutes. 0.3 g of L-cysteine ​​was added, the pH was adjusted to 5.0, and the mixture was stirred at 300 rpm for 3 hours to complete the cross-linking reaction between L-cysteine ​​and inulin to form thiolated inulin.

[0076] (3) Preparation of particle-filled microgels

[0077] 2.5g of thiolated inulin was dissolved in 100mL of PBS (pH 8) to form a thiolated inulin solution. 1.25g of quinoa starch-pomegranate polyphenol nanoparticles were then dispersed in the thiolated inulin solution. The solution was exposed to air at room temperature for 1 hour to complete the self-crosslinking of the free thiol groups to form a particle-filled microgel.

[0078] Example 3

[0079] (1) Preparation of quinoa starch-pomegranate polyphenol nanoparticles

[0080] 4g of quinoa starch was added to 100mL of ultrapure water to prepare a 4w / v% quinoa starch solution. 0.5g of pomegranate polyphenols was added to the solution and placed in an incubator with a magnetic field strength of 10T and stirred for 2 hours to obtain a quinoa starch-pomegranate polyphenol complex solution. The quinoa starch-pomegranate polyphenol complex solution was then adjusted to a pH of 7.5, and ultrasonic waves were applied at a power of 200W for 20 minutes to produce quinoa starch-pomegranate polyphenol nanoparticles.

[0081] (2) Preparation of thiolated inulin

[0082] 2.5 g of inulin was dissolved in 100 mL of sterile water. 0.6% (based on the mass of the inulin) of EDC and 0.6% (based on the mass of the inulin) of NHS were then added for activation and amidation of the inulin. The mixture was stirred at room temperature for 15 minutes. 0.2 g of L-cysteine ​​was added, the pH was adjusted to 5.0, and the mixture was stirred at 300 rpm for 3 hours to complete the cross-linking reaction between L-cysteine ​​and inulin to form thiolated inulin.

[0083] (3) Preparation of particle-filled microgels

[0084] 2.5g of thiolated inulin was dissolved in 100mL of PBS (pH 8) to form a thiolated inulin solution. 1.0g of quinoa starch-pomegranate polyphenol nanoparticles was dispersed in the thiolated inulin solution. The solution was exposed to air at room temperature for 1 hour to complete the self-crosslinking of the free thiol groups to form a particle-filled microgel.

[0085] Example 4

[0086] (1) Preparation of quinoa starch-pomegranate polyphenol nanoparticles

[0087] 4.5g of quinoa starch was added to 100mL of ultrapure water to prepare a 4.5w / v% quinoa starch solution. 0.75g of pomegranate polyphenols was added to the above solution, and the solution was placed in an incubator with a magnetic field strength of 15T and stirred for 3 hours to obtain a quinoa starch-pomegranate polyphenol complex solution. The quinoa starch-pomegranate polyphenol complex solution was then adjusted to a pH of 7.5, and ultrasonic waves were applied at a power of 300W for 30 minutes to obtain quinoa starch-pomegranate polyphenol nanoparticles.

[0088] (2) Preparation of thiolated inulin

[0089] 1.25 g of inulin was dissolved in 100 mL of sterile water. 0.3% (based on the mass of the inulin) of EDC and 0.3% (based on the mass of the inulin) of NHS were then added for activation and amidation of the inulin. The mixture was stirred at room temperature for 15 minutes. 0.15 g of L-cysteine ​​was added, the pH was adjusted to 5.0, and the mixture was stirred at 300 rpm for 3 hours to complete the cross-linking reaction between L-cysteine ​​and inulin to form thiolated inulin.

[0090] (3) Preparation of particle-filled microgels

[0091] 2.5g of thiolated inulin was dissolved in 100mL of PBS (pH 8) to form a thiolated inulin solution. 0.75g of quinoa starch-pomegranate polyphenol nanoparticles were dispersed in the thiolated inulin solution. The solution was exposed to air at room temperature for 1 hour to complete the self-crosslinking of the free thiol groups to form a particle-filled microgel.

[0092] Example 5

[0093] (1) Preparation of quinoa starch-pomegranate polyphenol nanoparticles

[0094] A 5 w / v quinoa starch solution was prepared by adding 5 g of quinoa starch to 100 mL of ultrapure water. 1 g of pomegranate polyphenols was added to the solution and stirred in an incubator at a magnetic field strength of 20 T for 4 hours to produce a quinoa starch-pomegranate polyphenol complex solution. The quinoa starch-pomegranate polyphenol complex solution was then adjusted to a pH of 7.5, and ultrasonic waves were applied at 400 W for 40 minutes to produce quinoa starch-pomegranate polyphenol nanoparticles.

[0095] (2) Preparation of thiolated inulin

[0096] 2.5 g of inulin was dissolved in 100 mL of sterile water. 0.25% (based on the mass of the inulin) of EDC and 0.25% (based on the mass of the inulin) of NHS were then added for activation and amidation of the inulin. The mixture was stirred at room temperature for 15 minutes. 0.6 g of L-cysteine ​​was added, the pH was adjusted to 5.0, and the mixture was stirred at 300 rpm for 3 hours to complete the cross-linking reaction between L-cysteine ​​and inulin to form thiolated inulin.

[0097] (3) Preparation of particle-filled microgels

[0098] 2.5g of thiolated inulin was dissolved in 100mL of PBS (pH 8) to form a thiolated inulin solution. 0.25g of quinoa starch-pomegranate polyphenol nanoparticles were dispersed in the thiolated inulin solution. The solution was exposed to air at room temperature for 1 hour to complete the self-crosslinking of the free thiol groups to form a particle-filled microgel.

[0099] Example 6

[0100] (1) Preparation of quinoa starch-pomegranate polyphenol nanoparticles

[0101] A 2.5 w / v quinoa starch solution was prepared by adding 2.5 g of quinoa starch to 100 mL of ultrapure water. 0.5 g of pomegranate polyphenols was added to the solution, which was then placed in an incubator with a magnetic field strength of 20 T and stirred for 4 hours to produce a quinoa starch-pomegranate polyphenol complex solution. The quinoa starch-pomegranate polyphenol complex solution was then adjusted to a pH of 6, and ultrasonic waves were applied at 400 W for 60 minutes to produce quinoa starch-pomegranate polyphenol nanoparticles.

[0102] (2) Preparation of thiolated inulin

[0103] 4g of inulin was dissolved in 100mL of sterile water. 1% (based on the weight of the inulin) of EDC and 1% (based on the weight of the inulin) of NHS were then added for activation and amidation of the inulin. The mixture was stirred at room temperature for 20 minutes. 0.2g of L-cysteine ​​was added, the pH was adjusted to 5.5, and the mixture was stirred at 200rpm for 2 hours to complete the cross-linking reaction between L-cysteine ​​and inulin to form thiolated inulin.

[0104] (3) Preparation of particle-filled microgels

[0105] 4g of thiolated inulin was dissolved in 100mL of PBS (pH 7) to form a thiolated inulin solution. 0.4g of quinoa starch-pomegranate polyphenol nanoparticles were then dispersed in the thiolated inulin solution. The solution was exposed to air at room temperature for 2 hours to complete the self-crosslinking of the free thiol groups to form a particle-filled microgel.

[0106] Example 7

[0107] (1) Preparation of quinoa starch-pomegranate polyphenol nanoparticles

[0108] A 2.5 w / v quinoa starch solution was prepared by adding 2.5 g of quinoa starch to 100 mL of ultrapure water. 0.5 g of pomegranate polyphenols was added to the solution, which was then placed in an incubator with a magnetic field strength of 20 T and stirred for 4 hours to produce a quinoa starch-pomegranate polyphenol complex solution. The quinoa starch-pomegranate polyphenol complex solution was then adjusted to a pH of 9, and ultrasonic waves were applied at 400 W for 60 minutes to produce quinoa starch-pomegranate polyphenol nanoparticles.

[0109] (2) Preparation of thiolated inulin

[0110] 5g of inulin was dissolved in 100mL of sterile water. 0.15% (based on the weight of the inulin) of EDC and 0.15% (based on the weight of the inulin) of NHS were then added for activation and amidation of the inulin. The mixture was stirred at room temperature for 30 minutes. 0.6g of L-cysteine ​​was added, the pH was adjusted to 6.0, and stirring was carried out at 500rpm for 2 hours to complete the cross-linking reaction between L-cysteine ​​and inulin to form thiolated inulin.

[0111] (3) Preparation of particle-filled microgels

[0112] 5g of thiolated inulin was dissolved in 100mL of PBS (pH 7.5) to form a thiolated inulin solution. 0.5g of quinoa starch-pomegranate polyphenol nanoparticles was then dispersed in the thiolated inulin solution. The solution was exposed to air at room temperature for 3 hours to complete the self-crosslinking of the free thiol groups to form a particle-filled microgel.

[0113] Comparative Example 1

[0114] Preparation of quinoa starch-pomegranate polyphenols complex

[0115] 5g of quinoa starch was added to 100mL of ultrapure water to prepare a 5w / v% quinoa starch solution. 1g of pomegranate polyphenol was added to the above solution, and the solution was placed in an incubator with a magnetic field strength of 15T and stirred for 5 hours to obtain a quinoa starch-pomegranate polyphenol complex solution.

[0116] Comparative Example 2

[0117] Preparation of quinoa starch / pomegranate polyphenol nanoparticles

[0118] A 5 w / v quinoa starch solution was prepared by adding 5 g of quinoa starch to 100 mL of ultrapure water. One g of pomegranate polyphenol was added to the solution and stirred for 1 hour to produce a quinoa starch / pomegranate polyphenol mixture solution. The quinoa starch / pomegranate polyphenol mixture solution was then adjusted to a pH of 7.5, and ultrasonic waves were applied at 500 W for 50 minutes to produce quinoa starch / pomegranate polyphenol nanoparticles.

[0119] Comparative Example 3

[0120] Preparation of quinoa starch-pomegranate polyphenol nanoparticles

[0121] A 5 w / v quinoa starch solution was prepared by adding 5 g of quinoa starch to 100 mL of ultrapure water. 1 g of pomegranate polyphenols was added to the solution and stirred in an incubator at a magnetic field strength of 15 T for 5 hours to produce a quinoa starch-pomegranate polyphenol complex solution. The quinoa starch-pomegranate polyphenol complex solution was then adjusted to a pH of 7.5, and ultrasonic waves were applied at 500 W for 50 minutes to produce quinoa starch-pomegranate polyphenol nanoparticles.

[0122] Animal experiments

[0123] Animal experiments were conducted using the dual-functional particle-filled microgel of Example 1, the quinoa starch-pomegranate polyphenols complex of Comparative Example 1, the quinoa starch / pomegranate polyphenols nanoparticles of Comparative Example 2, and the quinoa starch-pomegranate polyphenols nanoparticles of Comparative Example 3:

[0124] Treatment of colitis mice: mice injected with only water served as blank group (control), mice induced with only DSS (dextran sulfate sodium) (DSS) and mice treated with only pomegranate polyphenols (PP) served as control groups. Figures 1a to 2d .

[0125] like Figure 1a and Figure 1b As shown, the colon length of the mice treated with Example 1 was significantly longer than that of the control group, and the colon weight-to-length ratio of the mice treated with Example 1 was significantly lower than that of the control group, indicating that the dual-functional particle-filled microgel has a better therapeutic effect on ulcerative colitis.

[0126] like Figure 2a 、 Figure 2b 、 Figure 2c 、 Figure 2d As shown, the levels of pro-inflammatory cytokines (IL-1β, IL-6 and TNF-α) in the mice treated with Example 1 were significantly lower than those in the control group, and the level of anti-inflammatory cytokine (IL-10) was significantly higher than that in the control group, indicating that the dual-functional particle-filled microgel has a better therapeutic effect on ulcerative colitis.

[0127] Treatment of diabetic mice: The diabetic mice group injected with only water was used as the blank group (control1). Figure 3 .

[0128] like Figure 3 As shown, the area under the blood glucose curve (AUC) of the diabetic mice treated with Example 1 was significantly lower than that of the control group, indicating that the bifunctional particle-filled microgel can better lower blood glucose levels.

[0129] In addition, the inventors of this case also referred to the aforementioned embodiments and conducted experiments using other raw materials and conditions listed in this specification, and similarly produced dual-functional particle-filled microgels that can relieve colitis and diabetes, and achieved relatively ideal results.

[0130] The above-described embodiments merely illustrate several implementations of the present invention, and while their descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the patent. It should be noted that a person skilled in the art would be able to make numerous variations and improvements without departing from the spirit of the present invention, all of which fall within the scope of protection of the present invention. Therefore, the scope of protection of the patent for this invention shall be determined by the appended claims.

Claims

1. A method for preparing a bifunctional particle-filled microgel, characterized in that: include: Pomegranate polyphenols are mixed with a quinoa starch solution, subjected to a magnetic field treatment to obtain a quinoa starch-pomegranate polyphenol complex solution, the pH value of the quinoa starch-pomegranate polyphenol complex solution is adjusted to 6-9, and then subjected to ultrasonic treatment to obtain quinoa starch-pomegranate polyphenol nanoparticles; wherein the magnetic field intensity of the magnetic field treatment is 5-20T, the magnetic field treatment time is 1-5h; the ultrasonic power of the ultrasonic treatment is 100-500W, and the ultrasonic treatment time is 10-60min; Mixing inulin, water, 1-ethyl-(3-dimethylaminopropyl)carbodiimide, N-hydroxysuccinimide, and L-cysteine ​​for cross-linking reaction to obtain thiolated inulin; mixing the thiolated inulin with a phosphate buffer solution having a pH value of 7-8 to obtain a thiolated inulin solution; The quinoa starch-pomegranate polyphenol nanoparticles are mixed with a thiolated inulin solution and subjected to an oxidation reaction to obtain a bifunctional particle-filled microgel.

2. The preparation method according to claim 1, characterized in that The quinoa starch solution comprises quinoa starch and water, and the concentration of quinoa starch in the quinoa starch solution is 2.5-5 w / v%.

3. The preparation method according to claim 2, characterized in that The mass ratio of the pomegranate polyphenols to quinoa starch is 1:5-1:

20.

4. The preparation method according to claim 1, characterized in that include: The inulin aqueous solution, 1-ethyl-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide are mixed evenly, and then L-cysteine ​​is added to carry out a cross-linking reaction to obtain the thiolated inulin.

5. The preparation method according to claim 4, characterized in that The concentration of the inulin aqueous solution is 1-5 w / v%.

6. The preparation method according to claim 4, characterized in that The added amount of the 1-ethyl-(3-dimethylaminopropyl)carbodiimide is 0.1-1% of the mass of inulin.

7. The preparation method according to claim 4, characterized in that The added amount of the N-hydroxysuccinimide is 0.1-1% of the mass of inulin.

8. The preparation method according to claim 4, characterized in that The mass ratio of inulin to L-cysteine ​​is 20:1-2:

1.

9. The preparation method according to claim 4, characterized in that Specifically include: Inulin is dissolved in water, and then 1-ethyl-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide are added to obtain a mixture. The mixture is stirred at room temperature for 15-30 minutes, and then L-cysteine ​​is added. The pH value is adjusted to 5.0-6.0, and the mixture is stirred at room temperature at a speed of 200-500 rpm for 1-3 hours to complete the cross-linking reaction.

10. The preparation method according to claim 1, characterized in that include: The quinoa starch-pomegranate polyphenol nanoparticles are added to the thiolated inulin solution, and then reacted in air at room temperature for 1-3 hours to complete the self-crosslinking of the free thiol groups to obtain the bifunctional particle-filled microgel.

11. The preparation method according to claim 10, characterized in that: The mass ratio of the quinoa starch-pomegranate polyphenol nanoparticles to the thiolated inulin in the thiolated inulin solution is 1:2-1:

10.

12. The preparation method according to claim 10, characterized in that The concentration of thiolated inulin in the thiolated inulin solution is 2.5-5 w / v%.

13. A bifunctional particle-filled microgel prepared by the method according to any one of claims 1 to 12.

14. Use of the bifunctional particle-filled microgel according to claim 13 in the preparation of a drug for treating colitis or diabetes.

15. A drug for treating colitis or diabetes, characterized in that The invention comprises the bifunctional particle-filled microgel according to claim 13.

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