Preparation method for improving encapsulation efficiency of curcumin-millet prolamine nanoparticles
The preparation method of millet prolysin nanoparticles modified with hyaluronic acid has solved the problems of water solubility and stability of curcumin in the delivery field, achieving high encapsulation efficiency and drug loading, and broadening its application in the food and biomedical fields.
Patent Information
- Application Number
- CN202511256877.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-09-04
- Publication Date
- 2026-01-02
AI Technical Summary
The application of curcumin in the food and biomedical fields is limited by its poor water solubility, unstable structure, low bioavailability and poor permeability. In addition, the encapsulation rate of millet prolysin alone is low, resulting in the waste of bioactive substances.
Curcumin nanoparticles were prepared by antisolvent precipitation of millet prolysin nanoparticles modified with hyaluronic acid, and a hydrophilic layer was formed on the surface of the nanoparticles using hyaluronic acid to improve the encapsulation efficiency.
It significantly improved the encapsulation efficiency and drug loading of curcumin, enhanced its water solubility and chemical stability, and provided good biocompatibility and delivery performance.
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Abstract
Description
TECHNICAL FIELD
[0001] The application belongs to the technical field of biological agents, and specifically designs a hyaluronic acid modified curcumin-purothionin nanoparticle to improve the encapsulation efficiency and drug loading of curcumin. BACKGROUND
[0002] Curcumin is a polyphenolic substance derived from the rhizomes of Zingiberaceae or Costaceae plants, and is the main active ingredient of turmeric. Curcumin has various pharmacological activities: anti-inflammatory, antioxidant, anticancer, etc., but its poor water solubility, unstable structure, low bioavailability and poor permeability limit its application in food and biomedical fields.
[0003] Millet is a protein rich in proline, which is easy to interact with phenolic substances, so that purothionin can adsorb active substances into its hydrophobic core, thereby achieving effective protection and sustained release of drugs. At the same time, the high biocompatibility and biodegradability of purothionin make it have natural advantages in the application of food field. The gluten-free characteristics of millet are beneficial to its application in the intestinal sensitive population, which is of great significance to broaden the application field of the nanoparticle. However, the encapsulation efficiency of the delivery system using protein alone is low, which causes waste of bioactive substances. Hyaluronic acid, as an anionic polysaccharide, has a hydrophilic region composed of carboxyl and hydroxyl groups, a main chain and a partially hydrophobic region composed of acetyl groups. When combined with nanoparticles, hyaluronic acid forms a hydrophilic layer on the surface, thereby inhibiting the release of active substances and improving the encapsulation efficiency. In addition, hyaluronic acid has multiple functional groups, including carboxyl, amino, carboxylate of glucuronic acid, hydroxyl, N-acetylglucosamine at the reducing end, etc. These functional groups are conducive to the coupling of hyaluronic acid with active substances and proteins, and are conducive to improving the encapsulation efficiency of nanoparticles for active substances. This method has the advantages of simplicity and low cost, without the need for complex operating conditions and equipment, and the materials required are of natural origin, ensuring the safety of the nanoparticles, and is a promising protein modification technology. SUMMARY
[0004] In view of the problems and defects of curcumin in practical application, the purpose of the present application is to provide a preparation method of hyaluronic acid modified purothionin nanoparticles loaded with curcumin. On the one hand, the method uses anti-solvent precipitation to prepare nanoparticles loaded with curcumin to solve the difficulties of hydrophobic active substances in the delivery field; on the other hand, the nanoparticles are modified with hyaluronic acid to improve the encapsulation efficiency of the nanoparticles. This provides more solutions for food and health care fields and other fields.
[0005] To achieve the above object and other related objects, the technical scheme provided by the present application is as follows: a preparation method of curcumin-loaded zein-hyaluronic acid nanoparticles.
[0006] Step 1: Dissolve zein and curcumin in 70% ethanol solution, and magnetically stir to fully combine them.
[0007] Step 2: Dissolve hyaluronic acid in 70% ethanol solution.
[0008] Step 3: Add the complex of zein and curcumin to the ethanol solution of hyaluronic acid, and fully stir to make them interact.
[0009] Step 4: Add the complex of zein, curcumin and hyaluronic acid to 0 deionized water with pH 11.0.
[0010] Step 5: After centrifugation at 3000g for 10 minutes, collect the supernatant, which is the curcumin-loaded composite nanoparticles.
[0011] Further, the concentration of zein in the above technical step 1 is 20 mg / mL.
[0012] Further, the concentration of curcumin in the above technical step 1 is 0.8, 1, 1.5, 1.7 mg / mL.
[0013] Further, the concentration of hyaluronic acid in the above technical step 3 is 2 mg / mL.
[0014] Further, the ratio of solvent to antisolvent in the above technical step 3 is 1:4.
[0015] The principle of the present application is that by switching the solvent and antisolvent, the hydrophobic region of the protein is used as a carrier to form nanoparticles, and the hydrophilic part of the protein is exposed on the surface to prevent the aggregation of the nanoparticles by charge repulsion.
[0016] The advantages of the technical scheme of the present application are as follows: The present application uses a nano delivery carrier to deliver curcumin, which greatly improves the water solubility, chemical stability and biological efficacy of curcumin. The delivery carrier selected by the present application is zein from plant sources, and anionic polysaccharide hyaluronic acid is added to improve the stability of the nanoparticles. The prepared nanoparticles have good stability and biocompatibility, and provide a theoretical basis for the delivery of hydrophobic active substances represented by curcumin. BRIEF DESCRIPTION OF DRAWINGS
[0017] Figure 1 The particle size of the curcumin-zein-hyaluronic acid nanoparticles prepared by implementing steps 1-5 is measured. Figure 2 Preparation of the polydispersity index (PDI) of curcumin-prolamin-hyaluronic acid nanoparticles prepared in steps 1-5; Figure 3 Preparation of the zeta-potential of curcumin-prolamin-hyaluronic acid nanoparticles prepared in steps 1-5; Figure 4 Preparation of the encapsulation efficiency of curcumin-prolamin-hyaluronic acid nanoparticles prepared in steps 1-5; Figure 5 Preparation of the drug loading of curcumin-prolamin-hyaluronic acid nanoparticles prepared in steps 1-5.
[0018] Figure 6 Preparation of the Fourier infrared diffraction spectrum of curcumin-prolamin-hyaluronic acid nanoparticles prepared in steps 1-5. DETAILED DESCRIPTION
[0019] The following detailed description of the application is made with reference to the accompanying drawings. The embodiments of the application are implemented on the premise of the technical solutions of the application, and detailed implementation schemes and specific operation processes are given, but the protection scope of the application is not limited to the following embodiments.
[0020] According to the information contained in the present application, various changes to the precise description of the application can be easily made by those skilled in the art without departing from the spirit and scope of the appended claims. The examples described in the present application are only for professionals skilled in the art to understand and read, and are not intended to limit the defined conditions under which the present application can be implemented. Unless otherwise specified, the numerical parameters in the following examples of the present application are approximate values, which can be changed according to different ideal properties. Secondly, the experimental materials used in the following examples are purchased from conventional biochemical reagent stores unless otherwise specified.
[0021] Example 1: Preparation of curcumin-loaded prolamin-hyaluronic acid nanoparticles, comprising the following steps: (1) Weigh the prolamin of millet dispersed in 70% (v / v) ethanol, with a protein concentration of 20 mg / mL, and mix the mixture in a water bath at 60°C with magnetic stirring for 20 min. After cooling to room temperature, centrifuge the prolamin solution at 3000 g for 10 min, and take the supernatant for use. v / v
[0022] (2) Add curcumin (0.8 mg / mL) to the prolamin ethanol solution, mix the mixture at 600 rpm·min -1 Stir continuously for 1 hour to form a complex of curcumin and millet prolysin.
[0023] (3) The complex of curcumin and millet protein obtained in step (2) is added dropwise to an ethanol solution of hyaluronic acid (70%). v / v In the mixture, continue to stir magnetically for 1 hour.
[0024] (4) The mixture of curcumin, millet protein, and hyaluronic acid obtained in step (3) is added dropwise to deionized water at pH 11.0 and stirred for 30 min to obtain a millet protein-hyaluronic acid nano-mixture loaded with curcumin.
[0025] (5) Remove the ethanol from the nanoparticles obtained in step (4) by rotary evaporation and add the corresponding volume of deionized water. Then, centrifuge to remove the substances that do not form nanoparticles. The supernatant is the prepared curcumin-millet lysozyme-hyaluronic acid nanoparticles.
[0026] (6) The average particle size, PDI, and zeta potential of the prepared nanoparticles were determined using a nanoparticle size potentiometer. The encapsulation efficiency and drug loading were calculated using the following formula: W 1: The amount of curcumin added; W 2: Amount of unencapsulated curcumin; W 3: The total amount of all substances in the system; W 4: The total amount of particles that did not form nanoparticles. Example
[0027] A method for preparing hyaluronic acid-modified curcumin-loaded millet prolysin nanoparticles includes the following steps: (1) Weigh out millet prolysin and disperse it in 70% ( v / v The protein was dissolved in ethanol at a concentration of 20 mg / mL and the mixture was magnetically stirred in a water bath at 60 °C for 20 min. After cooling to room temperature, the millet alcohol-soluble protein solution was centrifuged at 3000 g for 10 minutes, and the supernatant was collected for later use.
[0028] (2) Add curcumin (1 mg / mL) to the ethanol solution of millet protein, and stir the mixture at 600 rpm·min. -1 Stir continuously for 1 hour to form a complex of curcumin and millet prolysin.
[0029] (3) The complex of curcumin and millet protein obtained in step (2) is added dropwise to an ethanol solution of hyaluronic acid (70%).v / v ) and the mixture was continuously stirred magnetically for 1 h.
[0030] (4) The mixture of curcumin, millet prolamine and hyaluronic acid obtained in step (3) was added dropwise into deionized water with pH 11.0 and stirred for 30 min to obtain the millet prolamine-hyaluronic acid nanocomposite loaded with curcumin.
[0031] (5) The nanoparticles obtained in step (4) were removed by rotary evaporation to remove ethanol, and a corresponding volume of deionized water was supplemented, followed by centrifugation to remove substances that did not form nanoparticles, and the supernatant was the prepared curcumin-millet prolamine-hyaluronic acid nanoparticles.
[0032] (6) The average particle size, PDI, zeta potential value, encapsulation efficiency and drug loading of the nanoparticles were determined according to the method in Example 1. Example
[0033] A preparation method of a hyaluronic acid modified millet prolamine nanocomposite loaded with curcumin, comprising the following steps: (1) Millet prolamine was weighed and dispersed in 70% (v / v) ethanol, wherein the protein concentration was 20 mg / mL, and the mixture was magnetically stirred in a water bath at 60°C for 20 min. After cooling to room temperature, the millet prolamine solution was centrifuged at 3000 g for 10 min, and the supernatant was taken for standby. v / v
[0034] (2) Curcumin (1.5 mg / mL) was added to the millet prolamine ethanol solution, and the mixture was continuously stirred at 600 rpm·min -1 for 1 h to form a complex of curcumin and millet prolamine.
[0035] (3) The complex of curcumin and millet prolamine obtained in step (2) was added dropwise into a hyaluronic acid ethanol solution (70%, v / v) and the mixture was continuously stirred magnetically for 1 h. v / v
[0036] (4) The mixture of curcumin, millet prolamine and hyaluronic acid obtained in step (3) was added dropwise into deionized water with pH 11.0 and stirred for 30 min to obtain the millet prolamine-hyaluronic acid nanocomposite loaded with curcumin.
[0037] (5) The nanoparticles obtained in step (4) were subjected to rotary evaporation to remove ethanol, and a corresponding volume of deionized water was added, followed by centrifugation to remove substances that did not form nanoparticles, and the supernatant was the prepared curcumin-millet prolamin-hyaluronic acid nanoparticles.
[0038] (6) The average particle size, PDI, zeta potential value, encapsulation efficiency and drug loading of the nanoparticles were determined according to the method in Example 1. Example
[0039] A preparation method of hyaluronic acid modified millet prolamin nanoparticles loaded with curcumin, comprising the following steps: (1) A certain amount of millet prolamin was dispersed in 70% (v / v) ethanol, and the protein concentration was 20 mg / mL. The mixture was magnetically stirred at 60°C for 20 min. After cooling to room temperature, the millet prolamin solution was centrifuged at 3000 g for 10 min, and the supernatant was reserved. v / v (2) Curcumin (1.7 mg / mL) was added to the ethanol solution of millet prolamin, and the mixture was continuously stirred at 600 rpm·min -1 for 1 h to form a complex of curcumin and millet prolamin.
[0040] (3) The complex of curcumin and millet prolamin obtained in step (2) was added dropwise to a hyaluronic acid ethanol solution (70%, v / v), and the mixture was continuously magnetically stirred for 1 h.
[0041] (4) The mixture of curcumin, millet prolamin and hyaluronic acid obtained in step (3) was added dropwise to deionized water with pH 11.0, and stirred for 30 min to obtain a mixture of millet prolamin-hyaluronic acid nanoparticles loaded with curcumin. v v (5) The nanoparticles obtained in step (4) were subjected to rotary evaporation to remove ethanol, and a corresponding volume of deionized water was added, followed by centrifugation to remove substances that did not form nanoparticles, and the supernatant was the prepared curcumin-millet prolamin-hyaluronic acid nanoparticles.
[0042] (6) The average particle size, PDI, zeta potential value, encapsulation efficiency and drug loading of the nanoparticles were determined according to the method in Example 1.
[0043] The nanoparticles in the above examples were structurally characterized, and the results are as follows:
[0044] Appendix
[0045] The nanoparticles in the above examples were structurally characterized, and the results are as follows: Appendix Figure 1 The particle size of the different nanoparticles obtained in Examples 1-4 is shown. With the increase of the concentration of curcumin, the particle size of the nanoparticles shows a trend of first decreasing and then increasing, and the particle size is in nanometer level, and the particle size is small.
[0046] The Figure 3 The zeta potential of the curcumin-pennisetum album prolamin-hyaluronic acid nanoparticles prepared in Examples 1-4 of the present application is shown. The absolute value of the zeta potential of the nanoparticles is higher than 30 mV, indicating that the nanoparticles have good stability.
[0047] The Figure 4 The encapsulation efficiency of the curcumin-pennisetum album prolamin-hyaluronic acid nanoparticles prepared in Examples 1-4 of the present application is shown. The encapsulation efficiency of the nanoparticles is higher than 80%, and the encapsulation efficiency of the nanoparticles is significantly improved compared with the nanoparticles of unmodified hyaluronic acid.
[0048] The Figure 5 The drug loading of the curcumin-pennisetum album prolamin-hyaluronic acid nanoparticles prepared in Examples 1-4 of the present application is shown. When the concentration of curcumin is 1.7 mg / mL, the drug loading of the nanoparticles is the highest.
[0049] The Figure 6 The Fourier infrared spectrum of the curcumin-pennisetum album prolamin-hyaluronic acid nanoparticles prepared in Examples 1-4 of the present application is shown. It is shown that curcumin is successfully loaded in the pennisetum album prolamin-hyaluronic acid nanoparticles, and the interaction force among the three is hydrogen bond and hydrophobic interaction.
[0050] It should be noted that the above examples are only used to illustrate the technical solutions of the present application, and the factual mode of the present application is not limited by the above examples. Those skilled in the art should understand that the present application can be modified or equivalently replaced without departing from the spirit and scope of the technical solutions of the present application, and all are included in the protection scope of the claims of the present application.
Claims
1. A process for the preparation of curcumin loaded zein-hyaluronic acid nanoparticles characterized in that The following steps: Step 1: Dissolve millet prolamin and curcumin in 70% ethanol solution, and magnetically stir to fully combine. Step 2: Dissolve hyaluronic acid in 70% ethanol solution. Step 3: Add the complex of millet prolamin and curcumin to the ethanol solution of hyaluronic acid, and fully stir to interact. Step 4: Add the complex of millet prolamin, curcumin and hyaluronic acid to deionized water at pH 11.
0. Step 5: After centrifugation at 3000g for 10 minutes, collect the supernatant, which is the curcumin-loaded complex nanoparticles.
2. The preparation method of curcumin-millet prolysin-hyaluronic acid nanoparticles according to claim 1, characterized in that... The concentration of millet prolamin in the above technical step 1 is 20 mg / mL.
3. The preparation method of curcumin-millet prolysin-hyaluronic acid nanoparticles according to claim 1, characterized in that... The concentration of curcumin in step 1 is 0.8-1.7 mg / mL.
4. The preparation method of curcumin-millet prolysin-hyaluronic acid nanoparticles according to claim 1, characterized in that... The concentration of hyaluronic acid in step 2 is 2 mg / mL.
5. The method for preparing curcumin-millet prolysin-hyaluronic acid nanoparticles according to claim 1, characterized in that... The ratio of solvent to anti-solvent in step 3 is 1:
4.
6. Curcumin-purothin-hyaluronic acid nanoparticle characterized in that: is prepared by the method of any one of claims 1-5.