Curcumin composite particles for improving alcohol-related liver diseases and preparation method of curcumin composite particles

Abstract

The invention provides curcumin composite particles for improving alcohol-related liver diseases and a preparation method of the curcumin composite particles, and belongs to the technical field of nutritional active ingredient delivery systems. According to the preparation method, zein is taken as a core carrier, curcumin encapsulation is realized through an anti-solvent precipitation method, a compact gel middle layer is constructed by further adopting the protection performance of carrageenan and combining calcium ion mediated ionic crosslinking, and then the outer layer is coated with galactosylated chitosan with liver targeting property through electrostatic adsorption, so that the curcumin-containing hydrogel is prepared. And the curcumin-zein-carrageenan-galactosylated chitosan composite particle with a core-shell-shell structure is innovatively constructed. Compared with free curcumin, the composite particle has obviously improved light and heat stability, realizes controlled release of curcumin in a gastrointestinal tract environment, exerts an effect of obviously improving the alcohol-related liver disease, and has a good application prospect in the aspect of development of functional products for preventing and treating the alcohol-related liver disease.

Description

Technical Field

[0001] This invention relates to the field of nutritional active ingredient delivery system technology, specifically to a curcumin complex particle for improving alcohol-related liver disease and its preparation method. Background Technology

[0002] Curcumin, a natural active ingredient extracted from the rhizome of turmeric, possesses various pharmacological activities, including antioxidant, anti-inflammatory, lipid-lowering, and anti-cancer effects, showing significant application potential in food additives and pharmaceutical preparations. However, its clinical application is significantly limited by its low water solubility, photothermal instability (susceptible to ultraviolet radiation and high temperatures), rapid metabolic rate, and low bioavailability. To address these shortcomings, researchers in recent years have constructed various delivery systems to improve the physicochemical stability of curcumin and enhance its in vivo delivery efficiency, thereby increasing its therapeutic efficacy.

[0003] With the rapid development of nanotechnology, nanomaterials have demonstrated significant application value in curcumin delivery systems due to their excellent biocompatibility, stability, and multifunctional design. Among them, zein, as a nanoparticle material, possesses multiple advantages. Its natural plant origin endows it with good biocompatibility, renewability, and environmental friendliness, meeting the requirements of green formulation development. Secondly, zein is rich in nonpolar amino acids, exhibiting significant hydrophobic properties, and can self-assemble into nanoparticles with controllable particle size through antisolvent precipitation, making it suitable for the efficient encapsulation of hydrophobic active ingredients. Although the stability of single zein nanoparticles to complex environments (such as pH and ionic strength) is limited, forming a core-shell structure by combining it with hydrophilic proteins or polysaccharides can significantly improve the mechanical strength and environmental tolerance of the particles. For example, carrageenan's molecular chains are rich in sulfate groups, which can form a three-dimensional network structure through electrostatic interactions or ionic cross-linking, enabling the controllable assembly of composite particles. Furthermore, the specific binding of calcium ions to the sulfate groups of carrageenan not only enhances the mechanical properties and thermal stability of the particles but also regulates drug release behavior. Combining calcium ion crosslinking strategies, carrageenan nanosystems have shown significant potential in delivering sensitive active ingredients and developing intelligent controlled-release formulations. Chitosan is the only natural polysaccharide with cationic properties. Galactosylated chitosan is a derivative obtained through the directional modification of chitosan molecules. Although some amino groups in galactosylated chitosan are replaced by galactose, the remaining amino groups, after protonation, still carry a positive charge. They can form a stable complex with the anionic polysaccharide carrageenan through electrostatic interactions. The sulfate groups of carrageenan synergistically form a dense network structure with calcium ions, serving as an intermediate layer to protect the core carrier. The outer galactosylated chitosan layer forms a protective shell through hydrogen bonding and electrostatic adsorption, which not only enhances the structural stability of the composite particles in the gastrointestinal environment but also has the ability to be specifically recognized by the liver. The liver surface contains a large number of asialoglycoprotein receptors (ASGPRs), which are expressed in very small amounts in other parts of the body. Galactose can specifically bind to ASGPRs to form receptor-ligand complexes. Galactosylated chitosan is specifically recognized by ASGPRs on the hepatocyte membrane, and galactosylated chitosan-modified particles carrying curcumin can increase the accumulation of curcumin in liver tumor tissues, achieving targeted delivery to the liver.

[0004] Although existing delivery strategies have improved the physicochemical stability and bioavailability of curcumin, its liver targeting efficiency and stability under complex physiological conditions still need to be optimized, and further research is needed to improve alcohol-related liver disease. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides a curcumin composite particle for improving alcohol-related liver disease and its preparation method. Based on the principle of complementary material properties, this invention innovatively constructs a curcumin composite particle with a core-shell-shell structure, significantly improving its physicochemical stability and bioavailability, thus enabling it to effectively improve alcohol-related liver disease.

[0006] To achieve the above objectives, the specific technical solution of the present invention is as follows:

[0007] In a first aspect, the present invention provides a method for preparing curcumin complex particles for improving alcohol-related liver disease, comprising the following steps:

[0008] (1) Curcumin and zein are dissolved together in ethanol to obtain a curcumin-zein solution. The curcumin-zein solution is then added dropwise to water to obtain a dispersion.

[0009] (2) Add the dispersion to the carrageenan aqueous solution, and then add calcium chloride aqueous solution dropwise to react and obtain a mixed solution;

[0010] (3) Add the mixed solution to the aqueous solution of galactosylated chitosan and remove the solvent to obtain curcumin-zein-carrageenan-galactosylated chitosan composite particles.

[0011] To address the shortcomings of existing technologies regarding the stability and liver targeting of curcumin, this invention, based on the principle of complementary material properties, uses zein as the core carrier and achieves curcumin encapsulation through antisolvent precipitation. Protective performance is optimized by adjusting the amount of carrageenan added. A dense gel interlayer is constructed using calcium ion-mediated ionic crosslinking, and galactosylated chitosan is then coated onto the outer layer via electrostatic adsorption. This innovatively constructs a core-shell-shell structured curcumin-zein-carrageenan-galactosylated chitosan composite particle. Through light / thermal stability tests, gastrointestinal simulated digestion models, and animal models of alcohol-related liver disease, the invention verifies that the curcumin-zein-carrageenan-galactosylated chitosan composite particle possesses excellent light / thermal stability, gastrointestinal digestive controlled-release performance, and efficacy in improving alcohol-related liver disease. This provides a safe and effective new intervention for alcohol-related liver disease and promotes the application of natural polyphenolic compounds in the field of liver disease improvement. In the method of this invention, the mass ratio of zein to carrageenan significantly affects the particle size, stability, and encapsulation efficiency of the composite particles. This invention controls the mass ratio of curcumin, zein, carrageenan, and galactosylated chitosan within the range of 10:100:(20-60):5, thereby enabling the composite particles to possess excellent physicochemical stability and liver-protective effects.

[0012] Furthermore, the mass ratio of curcumin, zein, carrageenan, and galactosylated chitosan is 10:100:(20-60):5.

[0013] Furthermore, the mass ratio of curcumin, zein, carrageenan, and galactosylated chitosan is 10:100:40:5.

[0014] Furthermore, in step (1), the dispersion is formed by continuous stirring with a constant temperature magnetic stirrer at a speed of 1500-2000 rpm for 10 min-1 h.

[0015] Further, in step (1), the volume ratio of the curcumin-zein solution to water is 1:3-5.

[0016] Further, in step (1), the concentration of zein in the curcumin-zein solution is 10-20 mg / mL.

[0017] Furthermore, in step (2), the reaction process is carried out by a constant temperature magnetic stirrer with a rotation speed of 1500-2000 rpm and a reaction time of 0.5-2 h.

[0018] Further, in step (2), the concentration of calcium ions in the mixed solution is 0.1-0.5 mmol / L.

[0019] Further, in step (2), the concentration of the carrageenan aqueous solution is 1-6 mg / mL.

[0020] Further, in step (3), the concentration of the galactosylated chitosan aqueous solution is 0.15-1 mg / mL.

[0021] Furthermore, in step (3), the solvent is removed by rotary evaporation at a temperature of 40°C and a pressure of -0.1 MPa.

[0022] Secondly, the present invention provides curcumin-zein-carrageenan-galactosylated chitosan composite particles prepared by the method described above.

[0023] Thirdly, the present invention provides the use of the curcumin-zein-carrageenan-galactosylated chitosan composite particles in the preparation of products (e.g., pharmaceuticals) that improve or treat alcohol-related liver disease.

[0024] Fourthly, the present invention provides a product (e.g., a pharmaceutical product) for improving or treating alcohol-related liver disease, comprising the aforementioned curcumin-zein-carrageenan-galactosylated chitosan composite particles.

[0025] Compared with the prior art, the advantages of the present invention are:

[0026] 1. This invention uses zein as the core carrier, encapsulates curcumin through antisolvent precipitation, optimizes the protective performance by adjusting the amount of carrageenan, constructs a dense gel interlayer by combining calcium ion-mediated ionic crosslinking, and then coats the outer layer with galactosylated chitosan through electrostatic adsorption. The final product is a curcumin-zein-carrageenan-galactosylated chitosan composite particle with a core-shell-shell hierarchical structure. Through light / thermal stability tests, gastrointestinal simulated digestion models, and animal models of alcohol-related liver disease, the invention verifies that the curcumin-zein-carrageenan-galactosylated chitosan composite particle significantly improves the water solubility and physicochemical stability of curcumin, reduces gastrointestinal metabolism of curcumin, and has a significant ameliorative effect on alcohol-related liver disease. This invention provides a safe and effective new intervention for alcohol-related liver disease and promotes the application of natural polyphenolic compounds in the field of liver disease improvement.

[0027] 2. The method provided by this invention has a simple process flow, is green and pollution-free, and has low cost, which is conducive to industrial production. Attached Figure Description

[0028] Figure 1 A schematic diagram of the preparation process of curcumin-zein-carrageenan-galactosylated chitosan composite particles for improving alcohol-related liver disease;

[0029] Figure 2 The average particle size and PDI test results of the composite particles prepared in Examples 1-6 are shown.

[0030] Figure 3 The zeta potential diagrams are shown for the composite particles prepared in Examples 1-6.

[0031] Figure 4 The encapsulation efficiency of curcumin in the composite particles prepared in Examples 1-6;

[0032] Figure 5 Field emission scanning electron microscope (FESEM) image of the composite particles prepared in Example 1;

[0033] Figure 6 Field emission scanning electron microscope image of the composite particles prepared in Example 4;

[0034] Figure 7 Field emission scanning electron microscope image of the composite particles prepared in Example 7;

[0035] Figure 8 Fourier transform infrared spectra of the composite particles prepared in Examples 1, 4, and 7 and the single materials in Comparative Examples 1-4;

[0036] Figure 9 The X-ray diffraction patterns are those of the composite particles prepared in Examples 1, 4, and 7 and the single materials in Comparative Examples 1-4.

[0037] Figure 10 The curcumin retention rates of the composite particles prepared in Examples 1, 4, and 7 and the single material in Comparative Example 1 after light treatment;

[0038] Figure 11 The curcumin retention rates of the composite particles prepared in Examples 1, 4, and 7 and the single material in Comparative Example 1 after heat treatment are shown.

[0039] Figure 12 The ratio of curcumin release rate during simulated gastrointestinal digestion is shown between the composite particles prepared in Examples 1, 4, and 7 and the single material in Comparative Example 1.

[0040] Figure 13 The body weight of the mice with alcohol-related liver disease in Example 8;

[0041] Figure 14 The liver index of the mice with alcohol-related liver disease in Example 8;

[0042] Figure 15 The transaminase levels in the serum of mice with alcohol-related liver disease in Example 8; wherein, Figure 15 A represents the level of AST; Figure 15 B represents the level of ALT;

[0043] Figure 16 The TG level in the serum of mice with alcohol-related liver disease in Example 8;

[0044] Figure 17 The total TC content in the serum of mice with alcohol-related liver disease in Example 8;

[0045] Figure 18 The HDL-C content in the serum of mice with alcohol-related liver disease in Example 8;

[0046] Figure 19 The LDL-C content in the serum of mice with alcohol-related liver disease in Example 8;

[0047] Figure 20 The levels of biomarkers of oxidative stress in the livers of mice with alcohol-related liver disease in Example 8; wherein, Figure 20 A represents the level of GSH-Px; Figure 20 B represents the level of SOD; Figure 20 C represents the level of GSH; Figure 20 D represents the level of MDA.

[0048] Different letters in the figure (e.g., a, b, c, d) indicate statistically significant differences (P < 0.05). Detailed Implementation

[0049] The technical solution of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0050] This invention provides a method for preparing curcumin complex particles that improve alcohol-related liver disease, such as... Figure 1 As shown, it includes the following steps:

[0051] (1) Curcumin and zein are dissolved together in ethanol to obtain a curcumin-zein solution. The curcumin-zein solution is then added dropwise to water to obtain a dispersion.

[0052] (2) Add the dispersion to the carrageenan aqueous solution, and then add calcium chloride aqueous solution dropwise to react and obtain a mixed solution;

[0053] (3) Add the mixed solution to the aqueous solution of galactosylated chitosan and remove the solvent to obtain curcumin-zein-carrageenan-galactosylated chitosan composite particles.

[0054] In some examples, the mass ratio of curcumin, zein, carrageenan, and galactosylated chitosan is 10:100:(20-60):5.

[0055] In some examples, in step (1), the dispersion is formed by continuous stirring with a constant temperature magnetic stirrer at a speed of 1500-2000 rpm for 10 min-1 h.

[0056] In some examples, in step (1), the volume ratio of the curcumin-zein solution to water is 1:3-5.

[0057] In some examples, in step (1), the concentration of zein in the curcumin-zein solution is 10-20 mg / mL.

[0058] In some examples, in step (2), the reaction process is carried out by a constant temperature magnetic stirrer with a speed of 1500-2000 rpm and a reaction time of 0.5-2 h.

[0059] In some examples, in step (2), the concentration of calcium ions in the mixed solution is 0.1-0.5 mmol / L; and the concentration of the carrageenan aqueous solution is 1-6 mg / mL.

[0060] In some examples, in step (3), the concentration of the galactosylated chitosan aqueous solution is 0.15-1 mg / mL.

[0061] Example 1

[0062] (1) Weigh 100 mg of zein and 10 mg of curcumin and dissolve them in 10 mL of 70% (v / v) ethanol aqueous solution. Seal the solution with plastic wrap and shield it from light. This solution is called solution one.

[0063] (2) Using a magnetic stirrer at 1600 rpm, solution one (10 mL) was added dropwise into 30 mL of ultrapure water and stirred continuously for 1 h. Then, it was placed in a rotary evaporator for rotary evaporation to remove ethanol (40 ℃, -0.1 MPa) to obtain curcumin-zein complex particles, named CZ.

[0064] Example 2

[0065] (1) Weigh 100 mg of zein and 10 mg of curcumin and dissolve them in 10 mL of 70% (v / v) ethanol solution. Seal the solution with plastic wrap and shield it from light. This solution is called Solution 1. Weigh 20 mg of carrageenan and dissolve it in 20 mL of ultrapure water. Seal the solution with plastic wrap. This solution is called Solution 2.

[0066] (2) Using a magnetic stirrer at 1600 rpm, solution one (10 mL) was added dropwise to 30 mL of ultrapure water at a constant speed and stirred for 10 min to obtain a dispersion. The dispersion was then injected into solution two (20 mL) at a constant speed and stirred for another 10 min. Then, 20 mL of 0.5 mmol / L calcium chloride aqueous solution was added dropwise and stirred for another 40 min to obtain a mixed solution. The mixed solution was then subjected to rotary evaporation in a rotary evaporator to remove ethanol (40℃, -0.1 MPa) to obtain curcumin-zein-carrageenan composite particles, named CZC20.

[0067] Example 3

[0068] The preparation method of the composite particles in this embodiment is basically the same as that in Example 2, except that the mass of carrageenan in this embodiment is 30 mg, and the resulting composite particles are named CZC30.

[0069] Example 4

[0070] The preparation method of the composite particles in this embodiment is basically the same as that in Example 2, except that the mass of carrageenan in this embodiment is 40 mg, and the resulting composite particles are named CZC40.

[0071] Example 5

[0072] The preparation method of the composite particles in this embodiment is basically the same as that in Example 2, except that the mass of carrageenan in this embodiment is 50 mg, and the resulting composite particles are named CZC50.

[0073] Example 6

[0074] The preparation method of the composite particles in this embodiment is basically the same as that in Example 2, except that the mass of carrageenan in this embodiment is 60 mg, and the resulting composite particles are named CZC60.

[0075] The composite particles prepared in Examples 1-6 were tested, and the specific test results are as follows:

[0076] (1) Determination of average particle size, polydispersity index (PDI), and zeta potential: The composite particle samples prepared in Examples 1-6 were placed in the sample cell (DTS0012), the temperature was set to 25 ℃, and the samples were measured three times consecutively. The average value of the results was taken. The composite particle samples prepared in Examples 1-6 were placed in the potential measurement cell (DTS1070), the equilibration time was 120 s, and the samples were measured three times consecutively, with an interval of 30 s between each measurement. The average value of the data was taken.

[0077] The average particle size and PDI test results of the composite particles prepared in Examples 1-6 are as follows: Figure 2 As shown in the figure, the particle size of CZ is 111.47±2.96 nm. After adding carrageenan, the particle size of CZC20 increases to 193.67±1.99 nm. Furthermore, the particle size of the composite particles gradually increases with the increase of carrageenan mass, indicating that carrageenan successfully adheres to the surface of CZ. The PDI of the composite particle dispersions prepared in Examples 1-6 is all below 0.5, indicating good stability of the composite particles. Compared with CZ, the PDI of CZC20, CZC30, and CZC40 is significantly lower. This may be because the positively charged zein and the anionic polysaccharide carrageenan combine through electrostatic interaction to form a more stable complex, significantly improving the monodispersity of the composite particles.

[0078] The zeta potential of the composite particles prepared in Examples 1-6 is as follows: Figure 3As shown in the figure, the zeta potential of CZ is +24.33±0.52 mV. After adding negatively charged carrageenan, the CZC composite particles (CZC20, CZC30, CZC40, CZC50, and CZC60) all exhibit anionic properties. Furthermore, the absolute value of the zeta potential of the CZC composite particles gradually increases with the increase of carrageenan mass. This result indicates that the stability of the composite particles increases with the increase of carrageenan mass.

[0079] (2) Determination of curcumin encapsulation efficiency: First, a standard curve of curcumin in ethanol-water solution was plotted. Next, the composite particle dispersions prepared in Examples 1-6 were centrifuged at low speed (1000 rpm, 10 min) to remove free curcumin. The supernatant was appropriately diluted, and the absorbance value was measured at 426 nm. The curcumin concentration was calculated by substituting it into the standard curve, and the mass of encapsulated curcumin was calculated. The curcumin encapsulation efficiency was calculated as follows:

[0080]

[0081] The encapsulation efficiency of curcumin in the composite particles prepared in Examples 1-6 is as follows: Figure 4 As shown in the figure, the encapsulation efficiency of CZ was only 34.16% ± 1.60%, indicating that a large amount of curcumin was not encapsulated in zein or that leakage occurred. After adding carrageenan, the encapsulation efficiency of curcumin significantly improved, with CZC20 reaching 85.55% ± 0.99%. This indicates that carrageenan promoted the encapsulation of curcumin by zein, possibly because the tight coating of carrageenan on the surface of zein helps zein capture curcumin or inhibits its escape, thus improving the encapsulation performance of zein. When the mass ratio of zein to carrageenan increased to 100:40, the encapsulation efficiency of CZC40 was 95.25% ± 1.04%. Further increases in the mass of carrageenan did not result in a statistically significant difference in the encapsulation efficiency of the resulting composite particles. In summary, a mass ratio of zein to carrageenan of 100:40 is most suitable for preparing CZC composite particles. In subsequent experiments and tests, the CZC used was specifically CZC40 composite particles.

[0082] Example 7

[0083] a. Weigh 100 mg of zein and 10 mg of curcumin and dissolve them in 10 mL of 70% (v / v) ethanol solution. Seal the solution with plastic wrap and protect it from light. This is called Solution 1. Weigh 40 mg of carrageenan and dissolve it in 20 mL of ultrapure water. Seal the solution with plastic wrap. This is called Solution 2. Weigh 5 mg of galactosylated chitosan and dissolve it in 30 mL of ultrapure water. Seal the solution with plastic wrap. This is called Solution 3. The preparation of Solutions 1, 2, and 3 is carried out by stirring continuously at 600 rpm for 10 h using a constant temperature magnetic stirrer.

[0084] b. Using a magnetic stirrer at 1600 rpm, 10 mL of Solution 1 was added dropwise to 30 mL of ultrapure water at a uniform rate, and the mixture was stirred continuously for 10 min to obtain a dispersion. The dispersion was then injected into 20 mL of Solution 2 at a uniform rate, and after stirring for 10 min, it was added dropwise to 20 mL of 0.5 mmol / L calcium chloride aqueous solution, and the mixture was stirred continuously for 10 min to obtain a mixed solution. The mixed solution was then slowly added dropwise to 30 mL of Solution 3, and the mixture was stirred continuously for 30 min. The mixture was then placed in a rotary evaporator for rotary evaporation to remove ethanol (40℃, -0.1 MPa) to obtain curcumin-zein-carrageenan-galactosylated chitosan composite particles, named CZCG.

[0085] The composite particles prepared in Examples 4 and 7 were tested. The tests included the determination of the average particle size, polydispersity index (PDI), and zeta potential of the composite particles, as well as the determination of the curcumin encapsulation efficiency. The test results are shown in Table 1.

[0086] Table 1: Particle size, PDI, zeta potential, encapsulation efficiency, and loading rate of the composite particles prepared in Examples 4 and 7

[0087]

[0088] In Table 1, * indicates 0.01 ≤ P < 0.05, ** indicates 0.001 ≤ P < 0.01, and *** indicates P < 0.001.

[0089] As shown in Table 1, the particle size and PDI of the CZCG composite particles modified with galactosylated chitosan were significantly increased compared to CZC, indicating that galactosylated chitosan successfully covered the surface of the CZC composite particles to form a complete protective shell. The zeta potential of the composite particles increased from -30.78 mV to -23.18 mV. This is because the protonated amino groups remaining in the galactosylated chitosan still carry a positive charge, which combines with the anionic polysaccharide carrageenan through electrostatic adsorption, neutralizing part of the charge. The encapsulation efficiency of curcumin in the CZCG composite particles increased significantly, indicating that the composite structure formed by galactosylated chitosan and carrageenan can more effectively load and protect curcumin.

[0090] The microstructure of the composite particles prepared in Examples 1, 4, and 7 was observed. The freeze-dried composite particle samples were adhered to conductive adhesive and subjected to platinum spraying treatment. The platinum spraying current was set to 30 mA, and the spraying time was 50 s. The microstructure of the samples was then observed using a field emission scanning electron microscope (FET), with an accelerating voltage of 3.0 kV and a magnification of 70 K.

[0091] Figure 5-7The images show field emission scanning electron microscope (FESEM) images of the microstructure of the composite particles CZ, CZC, and CZCG prepared in Examples 1, 4, and 7, respectively. As can be seen from the images, CZ exhibits significant aggregation and fails to maintain a clear structure. This may be because the surface of the simple zein nanoparticles contains a large number of nonpolar groups, generating strong hydrophobic attraction. In the aqueous interface, these groups lack stability and rapidly combine, resulting in aggregation. Conversely, CZC and CZCG exhibit obvious spherical characteristics, with relatively uniform size, dispersed distribution, and smooth surfaces. This indicates that the addition of carrageenan and galactosylated chitosan significantly improves the hydrophobicity of the zein surface and enhances the stability of the composite particles.

[0092] Comparative Example 1 is free curcumin, named Cur.

[0093] Comparative Example 2 is simple zein, named Zein.

[0094] Comparative Example 3 was pure carrageenan, named Car.

[0095] Comparative Example 4 is a simple galactosylated chitosan, named GC.

[0096] The composite particles prepared in Examples 1, 4, and 7 were compared with the single raw materials in Comparative Examples 1-4 for testing. The specific testing contents are as follows:

[0097] (1) Fourier transform infrared spectroscopy analysis: A certain mass of the sample to be tested was pressed into a potassium bromide plate, and then measured using a Fourier transform infrared spectrometer with a resolution of 2 cm⁻¹. -1 The scanning wavelength range is 4000-400 cm. -1 A total of 64 scans were performed.

[0098] Fourier transform infrared spectrum such as Figure 8As shown, comparing the infrared spectra of Curcumin and the three composite particles reveals that the characteristic peak of curcumin disappears in all three composite particles, indicating that the active ingredient curcumin has been successfully encapsulated within the composite particles. The characteristic absorption peaks of Zein are attributed to the vibration of the -OH group, amide I (stretching vibration of C=O), and amide II (CN stretching vibration and NH bending vibration), respectively. The characteristic peaks of amide I and amide II still exist in CZ, but their positions have changed, indicating that hydrophobic interactions are involved in the self-assembly process of zein nanoparticles. Similarly, the characteristic absorption peaks of amide I and amide II also shift in the CZC composite particles, indicating that the addition of carrageenan also leads to hydrophobic interactions. In the CZCG composite particles, the C=O and NH characteristic peaks of GC shift, indicating that the CZC composite particles bind to GC through electrostatic attraction to form CZCG. Furthermore, all three composite particles (CZ, CZC, and CZCG) show absorption bands caused by the stretching vibration of the -OH group, with changes in position, increased area, and increased intensity, indicating strong hydrogen bonding during the assembly of the composite particles.

[0099] (2) X-ray diffraction pattern analysis: Crystal diffraction patterns of different samples were obtained using an X-ray diffractometer. The detection parameters were as follows: accelerating voltage and tube current were 40 kV and 40 mA, respectively, and the 2θ angle scanning range was 4-60°. o The scanning interval was 5 seconds, and the scanning rate was 0.02. o / s.

[0100] X-ray diffraction pattern as follows Figure 9 As shown in the figure, Cur exhibits multiple strong and sharp diffraction peaks, indicating that Cur is a crystalline substance; Zein shows two broad and flat small diffraction peaks, Car shows no obvious diffraction peaks, and GC shows one small diffraction peak, indicating that Zein, Car, and GC are all amorphous. Observing the diffraction patterns of CZ, CZC, and CZCG reveals that the characteristic peaks of Cur no longer appear, indicating that Cur has lost its crystalline structure and is encapsulated in composite particles in an amorphous form.

[0101] The composite particles prepared in Examples 1, 4, and 7 were compared with the single material in Comparative Example 1 for testing. The specific testing results are as follows:

[0102] (1) Light and heat stability test: Cur, CZ, CZC, and CZCG were placed in a light box (35℃) at the same time, and the light intensity was set to 0.35 W / m 2Samples were subjected to light irradiation and collected at 30, 60, 90, 120, 150, and 180 mins. The curcumin retention was processed and measured, and the retention rate was calculated. Cur, CZ, CZC, and CZCG were simultaneously placed in a 90℃ water bath and heated in the dark. Samples were collected at 20, 40, 60, 80, 100, and 120 mins. The curcumin retention was processed and measured, and the retention rate was calculated.

[0103] The formula for calculating curcumin retention rate is as follows:

[0104]

[0105] Figure 10-11 The curcumin retention rates of Cur, CZ, CZC, and CZCG after light and heat treatments were shown. As can be seen from the figure, the curcumin retention rate decreased with the increase of light or heat treatment time. Among them, free curcumin had the worst light / thermal stability and the fastest degradation rate. Moreover, the curcumin retention rate increased with the increase of the composite particle shell layer.

[0106] (2) Simulated gastrointestinal digestion experiment: Simulated gastric juice was prepared by mixing 2.0 mg / mL sodium chloride and 3.2 mg / mL pepsin and adjusting the pH to 2.0. Simulated intestinal juice was prepared by mixing 12.0 mg / mL bile salts, 2.0 mg / mL trypsin, 6.8 mg / mL potassium dihydrogen phosphate, and 8.8 mg / mL sodium chloride. 10 mL of the composite particle or single material sample was mixed with an equal volume of simulated gastric juice and incubated in a 37℃ water bath shaker for 60 min. The pH was then adjusted to 7.0 with sodium hydroxide solution to stop gastric digestion, yielding the digestive fluid. The digestive fluid was then mixed with an equal volume of simulated gastric juice and incubated in a 37℃ water bath shaker for another 120 min. Samples were taken every 30 min during the entire simulated gastrointestinal digestion experiment to calculate the curcumin release rate.

[0107]

[0108] Figure 12The figure shows the curcumin release rates of Cur, CZ, CZC, and CZCG during simulated gastrointestinal digestion. As can be seen from the figure, free curcumin is largely digested by pepsin in the early stages of gastric digestion (0-30 min), with a release rate reaching 31.53% ± 2.91%, and this release rate remains higher than that of the CZ, CZC, and CZCG composite particles throughout the simulated gastrointestinal digestion process. The curcumin release rate decreases sequentially with increasing shell layer thickness of the composite particles. At 180 min, the curcumin release rates of the CZ, CZC, and CZCG composite particles were 66.09% ± 2.70%, 54.43% ± 2.86%, and 45.14% ± 3.86%, respectively. These results indicate that the protective layer formed by the carrier material reduces the direct contact between curcumin and digestive enzymes and decreases the influence of gastric acid on the active groups. This protective effect slows down the release and decomposition rate of curcumin while promoting its stable release and effective absorption in the intestine. In particular, galactosylated chitosan has adhesive properties, which can increase the contact area between the composite particles and the biofilm, thereby increasing the retention time of curcumin in the gastrointestinal tract and extending the target action time, thus greatly improving the bioavailability of curcumin.

[0109] Example 8

[0110] This embodiment uses mice as the research subject to investigate the ameliorative effect of the composite particles prepared in Examples 4 and 7 on alcohol-related liver disease.

[0111] Sixty 8-10 week old male C57BL / 6 mice were housed in a barrier environment at the Animal Experiment Center of Wuhan University. They were acclimatized for one week before the experiment. The mice were randomly divided into six groups of 10 mice each: control group, model group, carrier group, free curcumin group, CZC group, and CZCG group. Mice in the free curcumin, CZC group, and CZCG group were administered curcumin in different formulations (free curcumin, composite granules prepared in Example 4, and composite granules prepared in Example 7) by gavage once daily, with a dosage calculated at 25 mg / kg / day. The control and model groups were administered an equal volume of physiological saline by gavage. The carrier group was administered an equal volume of a zein-carrageenan-galactosylated chitosan (ZCC) carrier (preparation method as described in Example 7, except without curcumin) without curcumin. Except for the control group, all groups were simultaneously given a Lieber-DeCarli standard control liquid diet for 5 days of acclimatization, during which alcohol was gradually added. After the adaptation period, except for the control group, all groups were completely replaced with Lieber-DeCarli 5% (v / v) alcohol liquid diet for 10 days, and the diet was removed on the evening of the 10th day. After fasting for 6 hours, on the morning of the 11th day from 7 to 9 am, the model group, carrier group, free group, CZC group, and CZCG group were administered 35% (v / v) ethanol solution (5 g / kg) by gavage, while the control group was given an equal amount of maltodextrin with the same calories and volume. 9 hours later, the mice were anesthetized with isoflurane, the eyeballs were enucleated and blood was collected, and the mice were euthanized by cervical dislocation and the liver was harvested.

[0112] Changes in mouse body weight during the experiment, as follows Figure 13 As shown, the liver index of mice is as follows: Figure 14 As shown, the levels of transaminases (AST: aspartate aminotransferase; ALT: alanine aminotransferase), TG (triglycerides), TC (total cholesterol), HDL-C (high-density lipoprotein cholesterol), and LDL-C (low-density lipoprotein cholesterol) in mouse serum were as follows: Figure 15-19 As shown, the levels of markers of oxidative stress (GSH-Px: glutathione peroxidase; SOD: superoxide dismutase; GSH: glutathione; MDA: malondialdehyde) in mouse liver were detected as follows: Figure 20 As shown above, the experimental results indicate that the composite particles CZCG can slow down the weight loss in mice. Figure 13 ), and reduce liver index ( Figure 14 ), inhibit inflammatory damage ( Figure 15 ), regulate lipid metabolism disorders ( Figure 16-19 ), alleviate liver oxidative stress ( Figure 20 It showed significant improvement and therapeutic effects on alcohol-related liver disease.

[0113] In summary, this invention uses zein as the core carrier, encapsulates curcumin through antisolvent precipitation, and further utilizes the protective properties of carrageenan to construct a dense gel interlayer via calcium ion-mediated ionic cross-linking. Then, through electrostatic adsorption, a liver-targeting galactosylated chitosan is coated onto the outer layer, innovatively constructing a core-shell-shell structured curcumin-zein-carrageenan-galactosylated chitosan composite particle. Compared to free curcumin, the composite particle exhibits significantly improved light and thermal stability, enabling controlled release of curcumin in the gastrointestinal environment and significantly improving the effect of alcohol-related liver disease. This invention achieves efficient encapsulation and precise delivery of curcumin, showing promising application prospects in the development of functional products for the prevention and treatment of alcohol-related liver disease.

[0114] The above detailed embodiments describe the implementation of the present invention; however, the present invention is not limited to the specific details described in the above embodiments. Within the scope of the claims and technical concept of the present invention, various simple modifications and changes can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

Claims

1. A method for preparing curcumin complex particles for improving alcohol-related liver disease, characterized in that, Includes the following steps: (1) Curcumin and zein are dissolved together in ethanol to obtain a curcumin-zein solution. The curcumin-zein solution is then added dropwise to water to obtain a dispersion. (2) Add the dispersion to the carrageenan aqueous solution, and then add calcium chloride aqueous solution dropwise to react and obtain a mixed solution; (3) Add the mixed solution to the aqueous solution of galactosylated chitosan and remove the solvent to obtain curcumin-zein-carrageenan-galactosylated chitosan composite particles.

2. The method for preparing curcumin complex particles for improving alcohol-related liver disease according to claim 1, characterized in that, The mass ratio of curcumin, zein, carrageenan and galactosylated chitosan is 10:100:(20-60):

5.

3. The method for preparing curcumin complex particles for improving alcohol-related liver disease according to claim 2, characterized in that, The mass ratio of curcumin, zein, carrageenan, and galactosylated chitosan is 10:100:40:

5.

4. The method for preparing curcumin complex particles for improving alcohol-related liver disease according to claim 1, characterized in that, In step (2), the volume ratio of the dispersion to water is 1:3-5.

5. The method for preparing curcumin complex particles for improving alcohol-related liver disease according to claim 1, characterized in that, In step (2), the concentration of calcium ions in the mixed solution is 0.1-0.5 mmol / L.

6. Curcumin-zein-carrageenan-galactosylated chitosan composite particles prepared by the method described in any one of claims 1-5.

7. The use of the curcumin-zein-carrageenan-galactosylated chitosan composite particles according to claim 6 in the preparation of products for improving or treating alcohol-related liver disease.

8. A product for improving or treating alcohol-related liver disease, characterized in that, Including the curcumin-zein-carrageenan-galactosylated chitosan composite particles as described in claim 6.

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