Compound preparation containing curcumin nanoparticles as well as preparation method and application of compound preparation

Abstract

The invention provides a compound preparation containing curcumin nano-particles as well as a preparation method and application of the compound preparation. Curcumin is wrapped by alcohol-soluble protein to form nano-particles, and the nano-particles are used as a Pickering milk solid emulsifier for emulsifying and dispersing vitamin D and olive oil into an oil phase to form a stable droplet structure; furthermore, polysaccharide with enteric characteristics is used as a water-phase carrier material, and the emulsion is encapsulated in the pellet by using a hole sharpening-solidification method, so that the solidification of the liquid medicine (the emulsion) is realized; the three natural components, namely the curcumin, the vitamin D and the olive oil, are ingeniously integrated into a whole; the targeting property of the medicine at the colon part is improved; meanwhile, the destructive effect of the gastrointestinal tract environment on the medicine carrying nanoparticles is reduced.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of pharmaceutical preparations, and particularly relates to a compound preparation containing curcumin nanoparticles as well as a preparation method and application thereof BACKGROUND Inflammatory bowel disease (IBD) is a group of diseases that cause long-term inflammation of the colon and small intestine, including ulcerative colitis and Crohn's disease, which can further induce more serious and fatal diseases such as colorectal cancer. The main clinical medical interventions for IBD include aminosalicylates, antibiotics, corticosteroids and immunosuppressants. However, most of these drugs cannot address the root causes of IBD, such as intestinal mucosal damage, intestinal barrier dysfunction and intestinal flora imbalance. In addition, long-term use of these drugs can easily cause serious adverse events, including nausea, headache, acne, edema and nasopharyngitis. Current clinical treatment mainly focuses on symptomatic relief, and there is a lack of functional intervention means with anti-inflammatory, regulation of intestinal microenvironment and repair of intestinal barrier. Moreover, long-term medication is prone to produce tolerance and side effects, and it is urgent to develop safe and effective natural active ingredient preparations.

[0002] Curcumin, as a natural polyphenolic compound, is isolated from turmeric. Curcumin has been used as a traditional medicine for a long time to treat various health problems, especially for the gastrointestinal system. It can reduce intestinal inflammation by inhibiting inflammatory pathways such as NF-KB and MAPK, and also promote the proliferation of beneficial bacteria such as bifidobacteria and inhibit the growth of harmful bacteria such as Escherichia coli, thereby improving the imbalance of the intestinal flora in IBS patients. In addition, curcumin has antioxidant protection, improves the body's ability to scavenge oxygen free radicals, reduces oxidative stress damage to the intestinal tract, protects the intestinal mucosa from damage by active oxygen, and has therapeutic effects on bacterial, parasitic and fungal infections. Curcumin is a promising adjunctive therapy for gastrointestinal diseases, but its water solubility is poor, its chemical stability is low, it is rapidly metabolized in the gastrointestinal tract, and its bioavailability is low. Simple curcumin preparations have limited efficacy and need to rely on carrier technology to improve delivery efficiency.

[0003] Vitamin D is a fat-soluble steroid that is a key regulator of calcium and phosphorus metabolism. Studies have shown that vitamin D plays an important role in maintaining the integrity of the gastrointestinal barrier, monitoring the intestinal microbiota and inflammatory immune response. These mechanisms are important in preventing the development of IBD and improving disease symptoms.

[0004] Olive oil is rich in monounsaturated fatty acids, phenolic compounds (hydroxytyrosol, tyrosol, oleuropein, etc.), and other active ingredients. Related studies have confirmed that the effect of olive oil on patients with irritable bowel syndrome (IBS) can alleviate the core symptoms of IBS patients by reducing the abdominal visceral movement response and abdominal withdrawal reflex caused by colon-rectal distension, relieving abdominal pain and visceral hypersensitivity; up-regulating the expression of intestinal tight junction proteins and mucin, reducing intestinal permeability, and repairing damaged intestinal barrier; promoting the proliferation of beneficial bacteria such as bifidobacterium and lactobacillus, inhibiting harmful bacteria such as proteobacteria, regulating intestinal flora balance, and improving intestinal microenvironment. The effect of olive oil on IBS patients has the characteristics of multi-target and multi-pathway. Its natural active ingredients and mild mode of action make it suitable as a dietary ingredient for functional intervention of IBS. At the same time, as a natural oil carrier, olive oil can significantly improve the solubility of hydrophobic active ingredients such as curcumin and vitamin D. In cooperation with curcumin and vitamin D, it can further cover the multiple pathological links of IBS and improve the intervention effect.

[0005] The preparation of natural active ingredients such as curcumin, vitamins, and olive oil can achieve better results in the treatment of intestinal inflammation. However, curcumin has poor water solubility, is sensitive to light, has poor absorption, and has low bioavailability. It is not feasible to directly dissolve curcumin and vitamin D in olive oil.

[0006] Although there are methods of microencapsulation to improve curcumin in existing technologies, such as the invention patent "Method for preparing water-soluble curcumin by microencapsulation" with application number CN202310205336.0, which proposes a method of preparing a water phase with curcumin and chitosan, modified malt dextrin and water, an oil phase with triglyceride, mixing the water phase with the oil phase by shearing to obtain an emulsion, and then spray drying to obtain curcumin microcapsules, solving the problems of low encapsulation efficiency and poor stability of curcumin microcapsules. The invention patent "Preparation method of corn zein-sodium alginate composite film embedding curcumin" with publication number CN116554519A discloses the preparation of corn zein nanoparticles embedding curcumin by reverse solvent method before adding the antibacterial agent curcumin into the film-forming liquid, solving the problems of incompatibility between curcumin and water-soluble film-forming, and explosive release of curcumin in the film. However, the research mainly focuses on improving the physicochemical properties of curcumin; there is little attention on the composition and design of multi-component drug synergistic stable delivery system containing curcumin, which can achieve a higher application level. SUMMARY

[0007] To solve the problems in the prior art, the application provides a compound preparation containing curcumin nanoparticles, the compound preparation of the application fully plays the synergistic effect of the natural compound prolamine-coated curcumin nanoparticles, vitamin D and olive oil, improves the overall stability of the compound preparation, realizes the targeted delivery of the three active ingredients, can be applied to the preparation of a drug for treating inflammatory bowel disease, and has significant public health significance and social value.

[0008] To achieve the object of the application, the following technical solutions are provided. The application provides a compound preparation containing curcumin nanoparticles, and raw material components include prolamine-coated curcumin nanoparticles, vitamin D and olive oil.

[0009] The application further provides a preparation method of the compound preparation containing curcumin nanoparticles, the oil phase composed of vitamin D and olive oil is emulsified and dispersed into a uniform and stable emulsion by using the prolamine-coated curcumin nanoparticles as a Pickering solid emulsifier, and the emulsion is encapsulated to form the compound preparation in the form of micro-pellets.

[0010] Further, the preparation method of the compound preparation containing curcumin nanoparticles comprises the following steps. S1, vitamin D is dissolved in olive oil as an oil phase, a prolamine-coated curcumin nanoparticle solution is used as an aqueous phase, and after mixing, high-speed dispersion homogenization is performed to form a stable emulsion; S2, an enteric polysaccharide is dissolved in water, and the emulsion formed in step S1 is mixed uniformly at a volume ratio of 1:1, and the mixed liquid is dropped into a calcium chloride aqueous solution in the form of droplets to prepare curcumin-loaded micro-pellets.

[0011] Further, the oil phase and the aqueous phase in step S1 are mixed at a ratio of 1:5-20.

[0012] Further, the enteric polysaccharide in step S2 is sodium alginate or low-fat pectin.

[0013] Further, the preparation method of the prolamine-coated curcumin nanoparticles is as follows. (1) alcohol, prolamine and curcumin are added into ethanol, and stirring and dissolution are performed for standby use; (2) the mixed ethanol solution is subjected to rotary evaporation in a rotary evaporator to remove ethanol, and a thin film is formed on the inner wall of the rotary evaporation bottle; (3) PBS buffer is added to perform hydration treatment on the thin film formed on the inner wall of the container, and the thin film is dissolved by ultrasonic vibration hydration at 37 DEG C, and is sequentially filtered by polycarbonate membranes with pore sizes of 0.2 μm and 0.1 μm, respectively, to obtain prolamine-coated curcumin nanoparticles.

[0014] Further, the mass ratio of the prolamine to the curcumin is 10-50:1.

[0015] The application also provides application of the curcumin nanoparticle-containing compound preparation in preparation of a medicine for treating inflammatory bowel disease.

[0016] Compared with the prior art, the application has the following advantages: 1. The application creatively proposes a stable synergistic drug delivery system prepared by combining alcohol-soluble protein-wrapped curcumin nanoparticles, vitamin D and olive oil; the three natural compound components fully play the roles of anti-inflammation, intestinal barrier repair and improvement of intestinal microenvironment, solve the multiple pathological targets of IBS, realize multi-component delivery, provide a safe and effective new choice for functional intervention of IBS, do not introduce any irritating external chemical components, reduce the dependence of patients on chemical drugs, and have significant public health significance and social value.

[0017] 2. In the compound preparation, the olive oil serves as a natural oil phase carrier, vitamin D is dissolved in the olive oil, and the alcohol-soluble protein-wrapped curcumin nanoparticles not only improve the water solubility of curcumin, but also have better wrapping effect and targeted slow-release effect on curcumin, so that the drug property of curcumin is avoided from being released too early in the drug transportation process, and the drug efficacy is reduced; meanwhile, the alcohol-soluble protein-wrapped curcumin nanoparticle solution not only is an aqueous phase, but also serves as a Pickering solid emulsifier, can replace the additional introduction of irritating chemical surfactants, and enables the oil phase (olive oil) and the aqueous phase to be fully and uniformly mixed, so that the oil phase is emulsified and dispersed into stable small droplet structures, and the subsequent compound preparation in the form of micro-pellets is easier to dose and take, avoids the separation of the aqueous phase and the oil phase, and solves the problems of uneven content of active ingredients, unstable drug efficacy and the like after the preparation is prepared.

[0018] 3. The alcohol-soluble protein-wrapped curcumin nanoparticles are preferentially taken up by a large number of immune cells in the inflammation area, and can be transported through the gaps or small holes of the epithelial villus tips through the porous adsorption effect, so that the treatment purpose is improved.

[0019] 4. The polysaccharide having an enteric property is used as an aqueous carrier material, the emulsion is encapsulated in the micro-pellets by using the sharp-hole-solidification method, the solidification of the liquid drug (the above-mentioned emulsion) is realized, so that the targeting of the drug in the colon is improved, and the damage of the gastrointestinal environment to the drug-loaded nanoparticles is reduced; meanwhile, the overall stability of the compound preparation after solidification is improved. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 A morphology and particle size distribution diagram of the alcohol-soluble protein-wrapped curcumin nanoparticles prepared in Example 1; Figure 2 An in-vitro adhesion experiment and release curve diagram of the alcohol-soluble protein-wrapped curcumin nanoparticles prepared in Example 1; Figure 3 Appearance and droplet size of the alcohol-soluble protein-encapsulated curcumin nanoparticles prepared in Example 1 as Pickering emulsion solid emulsifier; Figure 4 Appearance and size of the wet pellets prepared in Example 2; Figure 5 In-vitro release process of the pellets prepared in Example 3; Figure 6 Active oxygen scavenging capacity of the compound preparation in Example 4; Figure 7 Therapeutic effect evaluation of the compound preparation in Example 5; Figure 8 Anti-inflammatory effect of the compound preparation in Example 5. DETAILED DESCRIPTION

[0021] The present application will be further described below in conjunction with the accompanying drawings and preferred embodiments, and the embodiments are only given to illustrate the present application, but not to limit the scope of the present application.

[0022] The materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

[0023] The experimental methods in the following examples are conventional methods unless otherwise specified.

[0024] Example 1 The present embodiment provides a preparation method of alcohol-soluble protein-encapsulated curcumin nanoparticles, which comprises the following steps: (1) 75 g of wheat alcohol-soluble protein and 5 g of curcumin are weighed and added into 100 mL of 70% ethanol, and stirred and dissolved for standby use; this embodiment only gives a ratio of the addition amount of alcohol-soluble protein and curcumin, and in addition to this, the mass ratio of alcohol-soluble protein and curcumin can be 10~50:1 to achieve the same technical effect, and the alcohol-soluble protein can also use corn alcohol-soluble protein and the like; (2) the solution prepared in step (1) is subjected to rotary evaporation in a rotary evaporator under the conditions of 45°C, -0.1 MPa and 85 r / min to remove ethanol, and a thin film is formed on the inner wall of the rotary evaporation bottle; (3) PBS buffer (pH=7.4) is added to perform hydration treatment on the thin film formed on the inner wall of the container, and the hydration is performed under ultrasonic wave (240 W, 40 kHz) oscillation at 37°C until the thin film is dissolved, and then the alcohol-soluble protein-encapsulated curcumin nanoparticles are obtained by sequentially filtering with polycarbonate membranes with pore sizes of 0.2 μm and 0.1 μm, respectively, each repeated for 3 times.

[0025] Quality evaluation of the alcohol-soluble protein-encapsulated curcumin nanoparticles prepared in the present embodiment (1) Content determination Chromatographic conditions and system suitability test: Wondasil C18 (4.6mm x 250mm, 5um) chromatographic column was used, with acetonitrile: 0.1% formic acid aqueous solution = 60:40 (volume ratio) mobile phase, flow rate 0.8mL·min -1 , detection wavelength 425 nm, column temperature 25℃, injection volume 20uL; 10ug·mL -1 of curcumin reference solution, curcumin nanoparticle test solution and blank nanoparticle test solution were prepared and analyzed by the above chromatographic conditions, chromatogram was recorded, peak area was calculated, and the theoretical plate number was 1543; the chromatogram could achieve baseline separation and the peak shape was stable; Linear relationship investigation: 1, 2, 3, 4, 5ml of curcumin reference stock solution was precisely measured and diluted to constant volume with methanol, and 10.0, 20.0, 30.0, 40.0, 50.0ug·mL -1 of series reference solution was obtained; injection analysis was carried out by the above chromatographic conditions, with mass concentration (C) as the abscissa and peak area (A) as the ordinate, linear regression was carried out, the regression equation was y=202.76x+2.1066, the correlation coefficient R 2 =0.9998 (n=5), the results showed that the mass concentration of curcumin was in the range of 10-50ug·mL -1 and the peak area showed good linear relationship.

[0026] Precisely take 1mL of curcumin nanoparticles, put it in a 10mL volumetric flask, add methanol ultrasonic (240W, 40kHz) treatment for 10min, dilute to the mark with methanol, shake well, and measure the peak area by the above chromatographic conditions.

[0027] (2) Determination of encapsulation efficiency and drug loading The encapsulation efficiency was measured by reverse dialysis method, the dialysis bag was filled with appropriate amount of purified water, tied tightly, and put into a beaker containing diluted nanoparticle solution, and placed in a shaking bed for dialysis, after equilibrium, the dialysate was measured by high performance liquid chromatography, and the content of the main drug was calculated, the encapsulation efficiency of the alcohol-soluble protein coated curcumin nanoparticles prepared in this example was 91.24%, and the drug loading was 4.73%.

[0028] (3) Determination of leakage rate A solution of curcumin nanoparticles encapsulated in alcohol-soluble protein was filled into ampoules and placed at room temperature (25 ℃) and refrigerated conditions (4 ℃). Samples were taken on days 0, 5, 15, and 30 to determine the amount of free drug before and after placement, and the leakage rate was calculated. The results showed that under refrigerated conditions (4 ℃), the leakage rate of curcumin nanoparticles was less than 1%, indicating good stability. However, at room temperature (25 ℃), the leakage rate gradually increased over time, and was less than 5% after 30 days.

[0029] (4) Particle size distribution and morphology Take an appropriate amount of lyophilized curcumin nanoparticle powder coated with alcohol-soluble protein, dilute it 5 times with purified water, mix well, filter through a 0.22µm microporous membrane, and use a laser particle size analyzer to determine the particle size and zeta potential of the nanoparticles. Figure 1 As shown, the nanoparticle dispersion was measured at 25 °C. The average particle size of the nanoparticles was 57.6 nm, and the polydispersity index was 0.19, indicating good dispersion. The zeta potential of the rice particles was -54.45 mV.

[0030] (5) In vitro adhesion test Take 10 mg of curcumin nanoparticles encapsulated in alcohol-soluble protein and place them in a solution containing 0.1 mg / mL. -1 Eosinophil cationic protein and transferrin were added to a PBS solution and shaken in a 37°C water bath at 100 r·min. -1 Samples were taken at 30, 60, 90, and 180 min, respectively, and subjected to 10000 r·min. -1 Centrifuge for 30 min, take an appropriate amount of the supernatant, treat with methanol, and then analyze by HPLC to calculate the adhesion rate. The results are as follows: Figure 2 As shown, the adhesion rate of curcumin nanoparticles gradually increased over time, reaching a maximum value of 95.5%, indicating that the nanoparticles have a good adhesion effect with cationic proteins.

[0031] (6) Release rate Take 2 mL of a curcumin nanoparticle dispersion containing a known amount of alcohol-soluble protein and place it into a pre-treated dialysis bag. Then, place the dialysis bag in 100 mL of release medium (0.9% NaCl solution) and shake it on a constant temperature shaker at (37.0±0.5) ℃ (100 r·min). -1 ), take 2 mL samples at regular intervals, and replenish with an equal volume of release medium at the same temperature. Filter the samples through a 0.22 μm microporous membrane, and perform the assay according to the method described in the content determination section. Calculate the cumulative release rate and plot the drug release curve. Figure 2 As shown, curcumin nanoparticles encapsulated by alcohol-soluble protein have sustained-release properties, with a cumulative release rate of 43.24% over 8 hours.

[0032] (7) Solubility determination Excess curcumin and lyophilized curcumin nanoparticles coated with prolyl protein were added to a certain amount of purified water and shaken in a 25 ℃ constant temperature water bath until equilibration. Afterward, appropriate samples were taken from each and placed in 10 mL volumetric flasks, diluted to volume with methanol. The solutions were then filtered through a 0.22 μm microporous membrane and analyzed according to the method described in the content determination section. The apparent solubility was calculated. The results showed that after preparation as nanoparticles, the solubility of curcumin increased from 0.126 mg / L. mL -1 Increased to 5.18mg mL -1 It increases by about 40 times.

[0033] (3) Using curcumin nanoparticles encapsulated with alcohol-soluble protein as a solid emulsifier in Pickering emulsion, such as Figure 3 As shown, the prepared Pickering emulsion solid emulsion is an orange-yellow water-in-oil emulsion with no oil droplets on the surface. The emulsion is thin and has strong fluidity. After dilution, the emulsion sample was observed under an optical microscope. The droplet size was less than 3 μm, and the outer layer of the oil droplet surface had an orange-yellow shadow, indicating that curcumin nanoparticles were attached to the surface of the oil droplets, thereby hindering the aggregation and coalescence of the droplets and contributing to the stability of the emulsion.

[0034] Example 2 This embodiment provides a compound preparation containing curcumin nanoparticles, the raw material composition of which includes curcumin nanoparticles encapsulated by alcohol-soluble protein, vitamin D and olive oil.

[0035] The preparation method of the compound preparation containing curcumin nanoparticles in this embodiment includes the following steps: S1. Dissolve vitamin D in olive oil as the oil phase and curcumin nano solution as the aqueous phase. Mix the two at a ratio of 1:5 (oil phase to water phase) and homogenize at 12000 r / min for 3 min using a high-speed dispersion homogenizer to form a stable emulsion. S2. Dissolve sodium alginate in water to prepare a 4% solution, mix it with the emulsion prepared in S1 at a volume ratio of 1:1, and drop the mixture into a 15% calcium chloride aqueous solution. After solidification, filter and dry to obtain curcumin-loaded microspheres.

[0036] Take a random number of the freshly prepared microspheres, absorb the surface moisture, and photograph them. Figure 4As shown, the wet microspheres have a particle size of about 2 mm, a smooth surface with an orange-yellow sheen, and are spherical in shape and uniform in size. One hundred dried microspheres were randomly selected and placed on glass slides in batches. The images were magnified and photographed using a Motic digital microscope. The particle size was measured one by one using Motic Image Plus 2.0 image analysis software (the scale was calibrated). The average value was calculated using Excel 2003 software. After fitting a log-normal distribution, D90, D50, and D10 were calculated. The span SD = (D90-D10) / D50 was calculated. The calculated average particle size of the microspheres was 1.23 ± 0.17 mm, and the span was 0.23 mm, verifying the accurate dosage of the microspheres prepared by this invention.

[0037] Example 3 This embodiment provides a compound preparation containing curcumin nanoparticles, the raw material composition of which includes curcumin nanoparticles encapsulated by alcohol-soluble protein, vitamin D and olive oil.

[0038] The preparation method of the compound preparation containing curcumin nanoparticles includes the following steps: S1. Vitamin D is dissolved in olive oil as the oil phase, and curcumin nano solution is the aqueous phase. The two are mixed at a ratio of 1:10 (oil phase to water phase) and homogenized at 12000 r / min for 3 min using a high-speed dispersion homogenizer to form a stable emulsion. S2. Dissolve low-fat pectin in water to prepare a 4% solution, mix it with the emulsion prepared in S1 at a volume ratio of 1:1, and drop the mixture into a 15% calcium chloride aqueous solution. After solidification, filter, and dry, curcumin-loaded microspheres are obtained.

[0039] The microspheres prepared in this embodiment have had their surface moisture removed, and a photograph is taken as follows. Figure 4 As shown, the wet microspheres have a particle size of about 2 mm, a smooth surface with an orange-yellow sheen, and are spherical in shape and uniform in size.

[0040] The microparticles prepared in this embodiment were subjected to an in vitro release experiment: an appropriate amount of microparticles were weighed and placed in a 37°C water bath constant temperature shaker, at 100 r·min. -1 The mixture was shaken, and the release media were simulated gastric fluid (pH=1.2) for 2 hours, simulated intestinal fluid (pH=6.8) for 3 hours, and simulated colonic fluid (pH=6) for 3 hours. Samples were taken periodically, filtered through a 0.45 μm microporous membrane, and the filtrate was treated with methanol before being injected for analysis under the above chromatographic conditions. The cumulative release of curcumin from the microcapsules was calculated. The results are as follows: Figure 5 As shown, curcumin microparticles release slowly in artificial gastric fluid, releasing only 10% in 2 hours. In artificial intestinal fluid, the cumulative release reaches more than 60%, while in colonic fluid, the release is complete, reaching about 95%.

[0041] Example 4 The compound preparation containing curcumin nanoparticles prepared in Example 2 was applied to scavenging reactive oxygen species (ROS) in mice. Twenty-five female C57BL / 6 mice (6-8 weeks old, weighing 18-22 g) were divided into five groups and allowed free access to food and water. After one week of acclimatization, the mice were administered PBS + water, PBS + 3% DSS, and compound preparations (high, medium, and low) + 3% DSS via gavage, respectively. Twelve hours after treatment on day 4, the mice were euthanized by cervical dislocation. Colon tissue was rapidly dissected and separated. The colon tissue was rinsed with pre-cooled PBS to remove surface blood and fat and connective tissue. The colon tissue was fixed in 4% paraformaldehyde for 4-6 hours, dehydrated using a gradient method, embedded, and prepared into 5 μm thick paraffin sections, which were then mounted on anti-detachment slides. The tissue sections were placed in a humidified chamber, and an appropriate amount of 20 μM DCFH-DA (2,-dichlorodihydrofluorescein diacetate) working solution was added. The sections were incubated at 37 ℃ for 30 min in the dark. After incubation, the sections were gently washed three times with PBS buffer, 5 ml each time. After removing unbound free probes, the sections were mounted and observed and analyzed using a Zeiss LSM 800 confocal microscope. The results showed that ROS levels were significantly increased in the colonic tissue of mice treated with DSS (PBS + 3% DSS), while ROS signal was significantly reduced in the colonic tissue of mice in the compound preparation + 3% DSS group. Figure 6 This indicates that the compound preparation can indeed effectively remove ROS.

[0042] Example 5 The curcumin-containing nanoparticle compound preparation prepared in Example 2 was used to treat DSS-induced IBD mice.

[0043] Fifteen female C57BL / 6 mice aged 6-8 weeks were selected and divided into three groups of five each. Before the experiment, the mice were allowed to acclimatize to their environment for one week. For the DSS-induced inflammatory bowel disease model, mice were given drinking water containing 3% DSS for 6 days, then switched to regular drinking water. For the TNBS- or oxazolone-induced inflammatory bowel disease model, mice were pre-sensitized on day 8 by injection of 150 μL of 1% TNBS or 3% oxazolone solution. Subsequently, on day 0, mice were given 100 μL of 2.5% TNBS or 1% oxazolone solution rectally. Healthy control mice received only regular drinking water. Afterwards, mice were administered phosphate-buffered saline (PBS) or the compound preparation via gavage at predetermined time points, with a dosage of 50 mg / kg (calculated as curcumin). d; Isopropyl-β-D-thiogalactopyranoside (IPTG) was added to the drinking water to induce the expression of catalase (CAT) and superoxide dismutase (SOD) in the gastrointestinal tract of mice. During the treatment, the weight of the mice was recorded daily. After the treatment, the mice were sacrificed and the distal colon tissue was collected for evaluation of various indicators such as hematoxylin-eosin staining, myeloperoxidase activity detection and inflammatory factor detection.

[0044] like Figure 7 As shown, the results indicated that, compared with the control group (PBS + water), mice in the DSS treatment group (PBS + 3% DSS) exhibited a range of IBD symptoms, such as weight loss, increased disease activity index (DAI), colon shortening and damage, and enhanced myeloperoxidase (MPO) activity, indicating that the C57BL / 6 mouse IBD model was successfully established. Compared with the control group (PBS + 3% DSS), the compound preparation treatment group significantly protected mice from DSS-induced IBD, including weight loss, colon shortening, and colon tissue damage.

[0045] The levels of typical pro-inflammatory cytokines, including interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6), were measured by ELISA; Figure 8 As shown, the levels of IL-1β, TNF-α, and IL-6 in the colon tissue of mice treated with DSS were significantly upregulated, indicating that the IBD model was successfully established. As expected, the levels of these pro-inflammatory cytokines in mice treated with the compound preparation were significantly reduced, further demonstrating that the compound preparation has excellent anti-inflammatory effects.

[0046] The above description is merely an embodiment of the present invention and does not limit the patent scope of the present invention. Any equivalent structural or procedural transformations made based on the content of the present invention specification, or direct or indirect applications in other related technical fields, are similarly included within the patent protection scope of the present invention.

Claims

1. A compound preparation containing curcumin nanoparticles, characterized in that: The ingredients include curcumin nanoparticles encapsulated in prolysin, vitamin D, and olive oil.

2. The method for preparing the compound preparation containing curcumin nanoparticles as described in claim 1, characterized in that: Curcumin nanoparticles encapsulated with alcohol-soluble protein are used as Pickering emulsion solids emulsifiers to emulsify and disperse an oil phase composed of vitamin D and olive oil to form a uniform and stable emulsion, and the emulsion is then encapsulated to form a micro-pellet compound preparation.

3. The method for preparing the compound preparation containing curcumin nanoparticles as described in claim 2, characterized in that, Includes the following steps; S1. Vitamin D is dissolved in olive oil as the oil phase, and curcumin nanoparticle solution encapsulated by alcohol-soluble protein is the aqueous phase. After the two are mixed, they are dispersed and homogenized at high speed to form a stable emulsion. S2. Dissolve the enteric polysaccharide in water and mix it with the emulsion prepared in step S1 at a volume ratio of 1:

1. Drop the mixture into the calcium chloride aqueous solution in droplets to obtain a compound preparation in the form of microspheres loaded with curcumin.

4. The method for preparing the compound preparation containing curcumin nanoparticles as described in claim 3, characterized in that: In step S1, the oil phase and the water phase are mixed in a ratio of 1:5 to 20.

5. The method for preparing the compound preparation containing curcumin nanoparticles as described in claim 3, characterized in that: In step S2, the enteric polysaccharide is sodium alginate or low-fat pectin.

6. The method for preparing the compound preparation containing curcumin nanoparticles as described in claim 2 or 3, characterized in that: The preparation method of the curcumin nanoparticles encapsulated by the alcohol-soluble protein is as follows: (1) Add alcohol-soluble protein and curcumin to ethanol, stir and dissolve for later use; (2) The mixed ethanol solution is placed in a rotary evaporator to remove the ethanol by rotary evaporation, and a thin film is formed on the inner wall of the rotary evaporation flask; (3) Add PBS buffer to hydrate the film formed on the inner wall of the container. Hydrate the film by ultrasonic oscillation at 37°C until the film dissolves. Filter the film through polycarbonate membranes with pore sizes of 0.2 μm and 0.1 μm respectively. Repeat the process multiple times to obtain curcumin nanoparticles encapsulated by alcohol-soluble protein.

7. The method for preparing the compound preparation containing curcumin nanoparticles as described in claim 6, characterized in that: The mass ratio of the prolysin to curcumin is 10~50:

1.

8. The application of the compound preparation containing curcumin nanoparticles as described in claim 1 in the preparation of a drug for treating inflammatory bowel disease.

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