Nanocurculigo peptides composition with anti-inflammatory efficacy and application

Abstract

The application provides a nano-edible bird's nest peptide composition with anti-inflammatory efficacy and an application thereof. The nano-edible bird's nest peptide composition comprises bird's nest peptides and at least one of green tea powder, needle cherry powder, red algae extract, ganoderma extract, ginkgo leaf extract and licorice extract. The bird's nest peptide composition has anti-inflammatory efficacy, good flavor and taste and good stability. The nano-particles and Pickering emulsion containing the bird's nest peptide composition have high stability, high water solubility, low wall stickiness, high bioavailability, high anti-inflammatory efficacy and good flavor and taste, and provide a new idea for industrial application of the bird's nest peptide and related products in the fields of food, medicine, skin care products and the like.

Description

Technical Field

[0001] This invention relates to the field of bird's nest peptides, and more specifically, to nano bird's nest peptide compositions with anti-inflammatory effects and their applications. Background Technology

[0002] With the fast pace of modern life and the development of the food industry, people consume more processed meats, refined grains, and sugary drinks, leading to increased inflammation levels in the body. These inflammatory responses, present in small amounts and for extended periods, result in a state of chronic inflammation. Chronic inflammation can damage tissues and organs, endanger cardiovascular health, and increase insulin resistance, among other things. Therefore, it is essential to incorporate high-quality anti-inflammatory foods, medications, and skincare products into people's daily lives.

[0003] Bird's nest is a traditional Chinese delicacy and a functional food with nourishing and medicinal properties, gaining increasing attention both in China and around the world. Products made from edible bird's nest through hydrolysis, with the main components having a relative molecular mass of less than 10,000, are called bird's nest peptides.

[0004] Currently, further research is needed on the anti-inflammatory effects of bird's nest peptides. Summary of the Invention

[0005] It should be noted that this invention is based on the following discoveries of the inventors:

[0006] Studies have found that bird's nest peptides possess certain anti-inflammatory effects. To further enhance the anti-inflammatory efficacy of their products, the inventors attempted to combine bird's nest peptides with other substances and studied the anti-inflammatory effects of these combinations, such as detecting changes in the expression levels of pro-inflammatory factors in animals after administration of the combination. During the research, it was discovered that, on the one hand, although some substances themselves possess certain anti-inflammatory effects, these effects were weakened when combined with bird's nest peptides, or when combined with other substances possessing similar anti-inflammatory properties. In other words, the combination of several substances with inherent anti-inflammatory effects did not achieve the expected enhanced anti-inflammatory effect. On the other hand, although one or more substances could synergistically enhance the anti-inflammatory effect with bird's nest peptides, the product's flavor and taste were unpleasant, making it difficult for the public to accept, and sometimes exhibiting poor stability. The inventors obtained a superior bird's nest peptide composition through screening: bird's nest peptides, green tea powder, acerola cherry powder, red algae extract, Ganoderma lucidum extract, Ginkgo biloba extract, and licorice extract. This composition can synergistically exert anti-inflammatory effects through multiple mechanisms. Simultaneously, the bird's nest peptide composition has a good flavor and taste.

[0007] Furthermore, the bioactive substances in the aforementioned bird's nest peptide composition, such as active polysaccharides, terpenoids, and phenolic compounds, are highly sensitive to environmental stresses caused by physical conditions (light, heat, etc.) and biochemical conditions (digestive enzymes, pH, etc.). Improper storage conditions and the digestive environment of the gastrointestinal tract can both lead to a decrease in the activity of these bioactive substances. At the same time, terpenoids and phenolic compounds have strong hydrophobicity, which is detrimental to their effective utilization in vivo.

[0008] Based on this, the inventors, through in-depth research, discovered that loading the aforementioned bird's nest peptide composition onto a carrier to form nanoparticles, due to the nanoparticles' large specific surface area and small particle size, can protect the bioactivity of the loaded bird's nest peptide composition, enhance its bioavailability and solubility, and achieve targeted release of the loaded bird's nest peptide composition in the intestine. Therefore, it can protect the bioactivity of the bird's nest peptide composition, enhance bioavailability, and achieve better anti-inflammatory effects. Furthermore, the bird's nest peptide nanoparticles can be formulated into a Pickering emulsion to improve their stability and further enhance their anti-inflammatory efficacy.

[0009] In view of this, in one aspect of the present invention, a bird's nest peptide composition is provided. According to an embodiment of the present invention, the bird's nest peptide composition comprises: bird's nest peptide and at least one of the following: green tea powder, acerola cherry powder, red algae extract, Ganoderma lucidum extract, ginkgo leaf extract, and licorice extract.

[0010] The bird's nest peptide composition of the present invention exhibits a synergistic effect of multiple substances, which can regulate the balance of cytokine secretion by modulating the NF-κB and MAPK signaling pathways, inhibit the synthesis of pro-inflammatory factors such as tumor necrosis factor (TNF-α) and interleukins IL-1β, IL-6, and IL-8, and upregulate the expression of anti-inflammatory factors IL-10 and TGF-β, thereby alleviating the inflammatory response induced by pro-inflammatory factors. It can also reduce the synthesis and release of NO and PGE2 by inhibiting the expression of iNOS and COX-2 metabolic enzymes. Different substances have different anti-inflammatory mechanisms; combining these substances can achieve multi-pathway anti-inflammatory effects and synergistic anti-inflammatory effects. Simultaneously, the above-mentioned bird's nest peptide composition has good stability and flavor, and is widely accepted by the public.

[0011] According to embodiments of the present invention, the above-mentioned bird's nest peptide composition may further have the following additional technical features:

[0012] According to embodiments of the present invention, the bird's nest peptide composition comprises: bird's nest peptide, green tea powder, acerola cherry powder, red algae extract, Ganoderma lucidum extract, ginkgo leaf extract, and licorice extract. These seven substances can synergistically exert anti-inflammatory effects, exhibit good stability and flavor, and are widely accepted by the public.

[0013] According to embodiments of the present invention, the bird's nest peptide composition comprises, by weight, 0.1-3 parts bird's nest peptide, 10-20 parts green tea powder, 1-10 parts acerola cherry powder, 0.1-1 parts red algae extract, 0.1-1 parts Ganoderma lucidum extract, 0.1-1 parts Ginkgo biloba extract, and 0.1-1 parts licorice extract. The inventors obtained the above-mentioned preferred composition ratio through extensive experiments, thereby further improving the anti-inflammatory efficacy, stability, and flavor of the bird's nest peptide composition.

[0014] In another aspect of the invention, a nanoparticle is provided. According to an embodiment of the invention, the nanoparticle comprises: the aforementioned bird's nest peptide composition; and a carrier on which the bird's nest peptide composition is loaded.

[0015] Because the bioactive substances in the bird's nest peptide composition, such as active polysaccharides, terpenoids, and phenolic compounds, are highly sensitive to environmental stresses caused by physical conditions (light, heat, etc.) and biochemical conditions (digestive enzymes, pH, etc.), improper storage conditions and the gastrointestinal digestive environment can both lead to a decrease in the activity of these bioactive substances. Furthermore, terpenoids and phenolic compounds have strong hydrophobicity, which is detrimental to their effective utilization in vivo.

[0016] Furthermore, the above composition is loaded onto a carrier to form nanoparticles. These bird's nest peptide nanoparticles exhibit good biocompatibility, protecting the bioactivity of the loaded bird's nest peptide composition, enhancing bioavailability and solubility, and achieving targeted release and delivery within the intestine. Therefore, they can protect the bioactivity of the bird's nest peptide composition, enhance bioavailability, and achieve better anti-inflammatory effects. Thus, the nanoparticles according to the embodiments of the present invention possess high stability, high water solubility, low wall adhesion, high bioavailability, high anti-inflammatory efficacy, and good flavor and taste, providing new ideas for the industrial application of bird's nest peptides and related products in the fields of food, pharmaceuticals, and skincare products.

[0017] According to embodiments of the present invention, the carrier comprises sodium tripolyphosphate and chitosan. The cationic polyelectrolyte properties of chitosan allow it to generate strong electrostatic interactions with mucus or negatively charged mucosal surfaces, enabling the nanoparticles to adhere to the small intestinal mucosal surface for extended periods and enhancing cell permeability, thereby improving the bioavailability of the composition. The phosphate groups in sodium tripolyphosphate can crosslink with chitosan to form nanoparticles.

[0018] According to an embodiment of the present invention, the mass ratio of sodium tripolyphosphate to chitosan is 1:3 to 1:7. This ensures sufficient phosphate groups to crosslink with chitosan, forming stable nanoparticles, while also preventing excessive chitosan molecules from crosslinking with sodium tripolyphosphate, forming more intermolecular hydrogen bonds, which would increase particle size and lead to precipitation.

[0019] According to an embodiment of the present invention, the mass ratio of the bird's nest peptide composition to chitosan is 1:3 to 1:6. Therefore, the bird's nest peptide composition can be effectively loaded onto a carrier formed by chitosan and sodium tripolyphosphate, thereby better exerting its anti-inflammatory effects.

[0020] According to an embodiment of the present invention, the chitosan has an acetylation degree of greater than 80% and a molecular weight of 50-100 kDa.

[0021] According to an embodiment of the present invention, the particle size of the nanoparticles is 150.5 to 201.0 nm.

[0022] According to embodiments of the present invention, the nanoparticles are provided in the form of a nanoparticle dispersion. Specifically, dissolving the nanoparticles in a solvent to form a nanoparticle dispersion facilitates the utilization of the nanoparticles, for example, in the preparation of Pickering emulsions. Exemplarily, the solvent includes water, etc.

[0023] In another aspect, the present invention provides a method for preparing the aforementioned nanoparticles. According to an embodiment of the present invention, the method includes: loading the aforementioned bird's nest peptide composition onto the carrier to obtain the nanoparticles. Thus, the nanoparticles prepared by the method according to the embodiment of the present invention possess high stability, high water solubility, low wall adhesion, high bioavailability, high anti-inflammatory efficacy, and good flavor and texture. The preparation method is simple and easy to operate, suitable for large-scale production.

[0024] According to an embodiment of the present invention, the method includes: (1) adding a solution containing the bird's nest peptide composition dropwise to a chitosan solution and stirring to obtain a mixture; (2) adding a sodium tripolyphosphate solution dropwise to the mixture and stirring to obtain the nanoparticles. The bird's nest peptide composition is first mixed with chitosan, and then crosslinked with chitosan under the action of sodium tripolyphosphate to form nanoparticles.

[0025] According to embodiments of the present invention, the concentration of the solution containing the bird's nest peptide composition is 1–10 mg / mL. The inventors obtained this preferred concentration through extensive experimentation, thereby enabling better encapsulation of the bird's nest peptide composition by the carrier.

[0026] According to an embodiment of the present invention, the concentration of the chitosan solution is 0.5–10 mg / mL. The inventors obtained the above-mentioned optimal chitosan solution concentration through numerous experiments. This concentration allows for the formation of nanoparticles while avoiding the formation of micron-sized particles due to excessive chitosan participation in the cross-linking reaction. Furthermore, the electrostatic repulsion is insufficient to maintain the stability of these large particles, thus preventing precipitation.

[0027] According to an embodiment of the present invention, the pH value of the chitosan solution is 4.0 to 5.0. This facilitates the protonation of chitosan and the cross-linking of phosphate groups in sodium tripolyphosphate with the amino groups of chitosan, forming nanoparticles.

[0028] According to an embodiment of the present invention, the chitosan solution contains acetic acid.

[0029] According to embodiments of the present invention, the concentration of the sodium tripolyphosphate solution is 0.5–20 mg / mL. The inventors obtained the above-mentioned preferred sodium tripolyphosphate solution concentration through numerous experiments. This concentration provides sufficient phosphate groups to crosslink with chitosan to form nanoparticles, while also preventing excessive crosslinking of sodium tripolyphosphate with chitosan, which would lead to more intermolecular hydrogen bond formation, increasing the nanoparticle size and causing precipitation.

[0030] According to an embodiment of the present invention, in step (1), the stirring time is 20 to 40 minutes; in step (2), the stirring time is 40 to 50 minutes. This ensures that the reaction occurs fully.

[0031] According to an embodiment of the present invention, in step (2), the nanoparticle suspension obtained by stirring is centrifuged, washed and dried to obtain nanoparticle powder.

[0032] In another aspect, the present invention provides a Pickering emulsion. According to an embodiment of the present invention, the Pickering emulsion comprises: a dispersion of the aforementioned nanoparticles and an oil phase. Thus, the Pickering emulsion according to an embodiment of the present invention exhibits high stability, high water solubility, low wall adhesion, high anti-inflammatory efficacy, and good flavor and mouthfeel.

[0033] According to an embodiment of the present invention, the mass ratio of the nanoparticle dispersion to the oil phase is (1-9):1, wherein the mass concentration of the nanoparticle dispersion is 0.1-5.0%. This forms a stable Pickering emulsion.

[0034] According to embodiments of the present invention, the oil phase includes at least one of the following: corn oil, palm oil, coconut oil, olive oil, fish oil, walnut oil, perilla oil, flaxseed oil, and chia seed oil.

[0035] In another aspect, the present invention provides a method for preparing the aforementioned Pickering emulsion. According to an embodiment of the present invention, the method includes: mixing a dispersion of the aforementioned nanoparticles with an oil phase to obtain the Pickering emulsion. Thus, the Pickering emulsion obtained by the method according to the embodiment of the present invention exhibits high stability, high water solubility, low wall adhesion, high anti-inflammatory efficacy, and good flavor and mouthfeel. The preparation method is simple and easy to operate, and suitable for large-scale production.

[0036] According to an embodiment of the present invention, the method includes: shearing and stirring the dispersion of the nanoparticles and the oil phase, and homogenizing the resulting mixture using a microfluidic jet to obtain the Pickering emulsion. This allows for better dispersion, homogenization, and emulsification. Exemplarily, a high-speed shear emulsifier can be used for shearing and stirring, and high-pressure microfluidic jet homogenization can be employed.

[0037] According to an embodiment of the present invention, the shearing and stirring speed is 8000–12000 rpm, and the time is 1–5 min. This allows for better performance.

[0038] In another aspect, the present invention provides a food, pharmaceutical, or skincare product. According to embodiments of the present invention, the food, pharmaceutical, or skincare product comprises at least one of the following: the aforementioned bird's nest peptide composition, the nanoparticles, and the Pickering emulsion. Therefore, the food, pharmaceutical, or skincare product according to embodiments of the present invention possesses advantages such as high anti-inflammatory efficacy, high stability, high water solubility, and low wall adhesion, and has high application value.

[0039] In another aspect of the invention, the invention proposes the use of at least one of the aforementioned bird's nest peptide composition, the nanoparticles, and the Pickering emulsion in the preparation of food, pharmaceutical, or skincare products. According to embodiments of the invention, the food, pharmaceutical, or skincare product is used for anti-inflammatory purposes.

[0040] Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. Attached Figure Description

[0041] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the description of the embodiments taken in conjunction with the following drawings, in which:

[0042] Figure 1 This diagram illustrates the NO content in a zebrafish according to an embodiment of the present invention.

[0043] Figure 2 A schematic diagram showing the IL-1 content in a zebrafish according to an embodiment of the present invention is shown;

[0044] Figure 3 A schematic diagram showing the IL-6 content in a zebrafish according to an embodiment of the present invention is shown;

[0045] Figure 4 A schematic diagram showing the TNF-α content in zebrafish according to an embodiment of the present invention is shown. Detailed Implementation

[0046] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention.

[0047] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Therefore, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. Furthermore, in the description of this invention, unless otherwise stated, "a plurality of" means two or more.

[0048] The endpoints and any values ​​of the ranges disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0049] In this document, the terms “comprising” or “including” are open-ended expressions, meaning they include the contents specified in this invention but do not exclude other aspects.

[0050] In this document, the terms “optionally,” “optionally,” or “optionally” generally refer to an event or condition that may, but may not, occur, and the description includes both cases in which the event or condition occurs and cases in which the event or condition does not occur.

[0051] In this article, the term "green tea powder" refers to a powdered food product made primarily from fresh green tea leaves, which can be obtained through self-preparation, gifting, or purchase. For example, the green tea powder is sourced from Shaanxi Jiahe Biotechnology Co., Ltd.

[0052] In this document, the term "acerola cherry powder" refers to a powdered food product made primarily from acerola cherry powder, which can be obtained through self-preparation, gifting, or purchase. For example, the acerola cherry powder is sourced from Xi'an Bingo Biochemical Technology Co., Ltd.

[0053] In the text, the term "red algae extract" refers to an extract obtained by extracting red algae as the main raw material, which comes from Shaanxi Sinote Biotechnology Co., Ltd.

[0054] In the text, the term "Ganoderma lucidum extract" refers to an extract obtained by extracting Ganoderma lucidum as the main raw material, which comes from Shaanxi Jiahe Biotechnology Co., Ltd.

[0055] In the text, the term "ginkgo leaf extract" refers to an extract obtained by extracting ginkgo leaves as the main raw material, which comes from Shaanxi Jiahe Biotechnology Co., Ltd.

[0056] In the text, the term "licorice extract" refers to an extract obtained by extracting licorice as the main raw material, which comes from Xinjiang Longhuiyuan Pharmaceutical Co., Ltd.

[0057] This invention proposes methods for preparing nanoparticles and Pickering emulsions, which may specifically include the following steps:

[0058] (1) Mixing: Mix bird's nest peptide with green tea powder, acerola cherry powder, red algae extract, Ganoderma lucidum extract, ginkgo leaf extract and licorice extract in the following proportions;

[0059]

[0060]

[0061] (2) Composition dissolution: Add purified water to the material obtained in step (1) to prepare a solution with a mass concentration of 1 to 10 mg / mL, and stir evenly.

[0062] (3) Preparation of chitosan solution: At room temperature, add chitosan to a 0.5% to 2.0% acetic acid solution and stir magnetically for 8 to 12 hours to obtain a clear solution, so that the final mass concentration of chitosan is 0.5 to 10 mg / mL. Filter the solution with a 0.45 μm microporous membrane.

[0063] (4) Adjust pH: Adjust the pH of the chitosan solution obtained in step (3) to 4.0-5.0 with 0.1M sodium hydroxide solution.

[0064] (5) Add the composition solution with a mass concentration of 1 to 10 mg / mL dropwise to the chitosan solution obtained in step (4) so ​​that the final mass ratio of the composition to chitosan is 1:(3 to 6), and stir magnetically for 30 min at room temperature.

[0065] (6) A sodium tripolyphosphate solution with a mass concentration of 0.5 to 2.0 mg / mL is slowly added dropwise to the mixed solution obtained in step (5) so that the final mass ratio of sodium tripolyphosphate to chitosan is 1: (3 to 7), and the mixture is magnetically stirred at room temperature for 45 min to obtain a suspension of bird's nest peptide composition nanoparticles.

[0066] (7) Centrifuge the bird's nest peptide composition nanoparticle suspension, wash the precipitate with distilled water, freeze dry, and obtain bird's nest peptide composition nanoparticle powder.

[0067] (8) The bird's nest peptide nanoparticle powder was resuspended in purified water to obtain a bird's nest peptide nanoparticle dispersion with a mass concentration of 0.1% to 5.0%.

[0068] (9) Mix the bird's nest peptide composition nanoparticle dispersion with oil at a mass ratio of (1-9):1, stir and disperse it for 1-5 minutes at a speed of 8000-12000 rpm using a high-speed shear emulsifier, and then pass it through a high-pressure microjet to obtain a stable Pickering emulsion of bird's nest peptide composition nanoparticles.

[0069] The present invention will be explained below with reference to embodiments. Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of the invention. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products. In the following embodiments, the bird's nest peptide is from Xiamen Yanzhiwu Siyong Biotechnology Co., Ltd., the green tea powder is from Shaanxi Jiahe Biotechnology Co., Ltd., the acerola cherry powder is from Xi'an Bingo Biochemical Technology Co., Ltd., the red algae extract is from Shaanxi Snowt Biotechnology Co., Ltd., the Ganoderma lucidum extract is from Shaanxi Jiahe Biotechnology Co., Ltd., the Ginkgo biloba extract is from Shaanxi Jiahe Biotechnology Co., Ltd., and the licorice extract is from Xinjiang Longhuiyuan Pharmaceutical Co., Ltd.

[0070] In the comparative examples below, the bird's nest peptide is from Xiamen Yanzhiwu Siyong Biotechnology Co., Ltd., the soy milk powder is from Joyoung Co., Ltd., the spinach powder is from Jiangsu Runhui Food Co., Ltd., the aloe vera extract is from Anhui Bensentang Biotechnology Co., Ltd., the chili extract is from Hunan Langlin Biological Resources Co., Ltd., the cinnamon powder is from Xi'an Lvtian Biotechnology Co., Ltd., and the garlic extract is from Hunan Langlin Biological Resources Co., Ltd.

[0071] Examples 1-5

[0072] In this embodiment, the bird's nest peptide composition was prepared according to the following method:

[0073] 1. The raw material composition of Examples 1-5 is shown in Table 1 by weight.

[0074] Table 1 Raw materials for bird's nest peptide composition

[0075]

[0076] 2. Steps:

[0077] Mix the raw materials in the specified proportions and stir until homogeneous to obtain the bird's nest peptide composition.

[0078] Example 6

[0079] Using the bird's nest peptide composition prepared in Example 5 as a raw material, bird's nest peptide composition nanoparticles with anti-inflammatory effects were prepared. The specific operation steps are as follows:

[0080] The bird's nest peptide composition was dissolved in purified water to prepare a 1.0 mg / mL solution and stirred evenly. At room temperature, chitosan was added to a 1.0% acetic acid solution to prepare a 1.0 mg / mL solution. After magnetic stirring for 12 h, the solution was filtered through a 0.45 μm microporous membrane. The pH of the chitosan solution was adjusted to 4.0, and the bird's nest peptide composition was added dropwise to the chitosan solution until the mass ratio of bird's nest peptide composition to chitosan was 1:4. The solution was magnetically stirred for 30 min at room temperature. Subsequently, a 0.5 mg / mL sodium tripolyphosphate solution was added dropwise to the above solution until the mass ratio of sodium tripolyphosphate to chitosan was 1:5. The solution was magnetically stirred for 45 min at room temperature. The product was centrifuged at 10000 rpm for 10 min, and the precipitate was washed with distilled water and lyophilized to obtain bird's nest peptide composition nanoparticle powder.

[0081] Examples 7-8

[0082] The preparation methods of the bird's nest peptide composition nanoparticles in Examples 7-8 are basically similar to those in Example 6. The main difference is that the mass concentration of the bird's nest peptide composition, the mass concentration of chitosan, and the mass ratio of the bird's nest peptide composition to chitosan are shown in Table 2.

[0083] Table 2 Preparation conditions of bird's nest peptide composition nanoparticles

[0084]

[0085] Example 9

[0086] Using the bird's nest peptide composition nanoparticles prepared in Example 8 as raw materials, a nanoparticle-stabilized Pickering emulsion was prepared. The specific operating steps are as follows:

[0087] Bird's nest peptide nanoparticle powder was resuspended in purified water to obtain a bird's nest peptide nanoparticle dispersion with a mass fraction of 1.0%. The bird's nest peptide nanoparticle dispersion was mixed with fish oil at a mass ratio of 4:1. After stirring and dispersing for 2 minutes at 12000 rpm using a high-speed shear emulsifier, the mixture was passed through a high-pressure microjet to obtain a stable Pickering emulsion of bird's nest peptide nanoparticles.

[0088] Examples 10-12

[0089] The preparation methods of the Pickering emulsion stabilized by the bird's nest peptide composition nanoparticles in Examples 10-12 are basically similar to those in Example 9. The main differences are as follows: the mass concentration of the bird's nest peptide composition nanoparticles, the mass ratio of the bird's nest peptide composition nanoparticle dispersion to fish oil, and the type of oil are shown in Table 3.

[0090] Table 3 Pickering Emulsion Preparation Conditions

[0091]

[0092] Comparative Example

[0093] In this comparative example, the bird's nest peptide composition was prepared according to the following method:

[0094] 1. The raw material components in the comparative proportion are shown below in parts by weight.

[0095] Table 4 Comparative Example Raw Material Composition

[0096]

[0097] 2. Steps:

[0098] Mix the raw materials in the specified proportions and stir until homogeneous to obtain the bird's nest peptide composition.

[0099] Experimental Example 1

[0100] The particle size changes of the bird's nest peptide composition nanoparticles and Pickering emulsions of Examples 6-12 were measured using a particle size analyzer over 21 days to characterize the storage stability of the bird's nest peptide composition nanoparticles and their Pickering emulsions.

[0101] As shown in Table 5, the bird's nest peptide composition nanoparticles prepared in this invention are all nanoscale, indicating successful preparation of nanoparticles. Different preparation conditions resulted in different particle sizes for the bird's nest peptide composition nanoparticles, but all were within 200 nm. Furthermore, during the 0–21 days of the experiment, the particle size of the bird's nest peptide composition nanoparticles and its Pickering emulsion did not significantly increase, nor did they exhibit significant aggregation, indicating that the bird's nest peptide composition nanoparticles and their Pickering emulsion have good stability, thus contributing to targeted release and improved bioavailability. The smallest nanoparticle size was observed when the mass concentration of the bird's nest peptide composition was 1.0 mg / mL, the chitosan mass concentration was 1.0 mg / mL, and the mass ratio of the bird's nest peptide composition to chitosan was 1:4. The Pickering emulsion formed when the mass fraction of the bird's nest peptide composition nanoparticles was 2.0% and the mass ratio of the bird's nest peptide composition nanoparticle dispersion to fish oil was 7:3.

[0102] Table 5. Particle size of bird's nest peptide composition nanoparticles

[0103]

[0104]

[0105] Experimental Example 2

[0106] The anti-inflammatory effects of bird's nest peptide composition, bird's nest peptide nanoparticles, and their stabilized Pickering emulsion were evaluated using a zebrafish model. The levels of nitric oxide (NO), interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in zebrafish treated with the bird's nest peptide composition, bird's nest peptide nanoparticles, and their stabilized Pickering emulsion were determined. The specific steps are as follows:

[0107] (1) Determination of NO content in zebrafish: Wild-type zebrafish at 7-9 hpf were collected into different test sample solutions of the same concentration and pretreated for 1 hour. Then, zebrafish larvae were transferred to 6-well plates, lipopolysaccharide was added, and the larvae were incubated with the corresponding test sample solutions for 24 hours. Afterward, the zebrafish larvae were washed with culture water and incubated at a constant temperature until they reached 72 hpf. Fluorescent probe staining solution DAF-FM DA was added to the zebrafish culture water, and the plates were incubated at 28.5℃ in the dark for 1 hour. After incubation, the zebrafish embryos were thoroughly washed with culture water, transferred to 96-well plates, and anesthetized with 0.04% tricaine solution. The embryos were observed under a microscope, and the fluorescence intensity was quantified using ImageJ software to detect NO content.

[0108] (2) Determination of IL-1, IL-6 and TNF-α content in zebrafish: 24 hpf zebrafish were placed in 6-well plates, and different test sample solutions and lipopolysaccharide of the same concentration were added to each group. Lipopolysaccharide was used as a positive control group. The zebrafish were incubated at constant temperature until 72 hdf, ground and centrifuged at 4℃. The supernatant was collected and the content of inflammatory factors IL-1, IL-6 and TNF-α in the fish was determined according to the ELISA kit instructions.

[0109] Conclusion: When inflammation occurs in the body, the levels of pro-inflammatory factors IL-1, IL-6, and TNF-α are abnormally elevated, and macrophages release large amounts of NO in the early stages of inflammation. Therefore, NO levels and the levels of pro-inflammatory factors IL-1, IL-6, and TNF-α are considered important markers of the severity of inflammation. Figure 1-4 As shown, the bird's nest peptide composition, bird's nest peptide nanoparticles, and their stable Pickering emulsion in the comparative and examples all reduced the NO content and the content of pro-inflammatory factors IL-1, IL-6, and TNF-α in the inflamed zebrafish to varying degrees, indicating that the bird's nest peptide composition, bird's nest peptide nanoparticles, and their stable Pickering emulsion all have excellent anti-inflammatory activity. Legumes, spinach, aloe vera, chili peppers, cinnamon, and garlic are also recognized anti-inflammatory foods with good anti-inflammatory effects. However, after being combined with bird's nest peptides, the NO content and the content of pro-inflammatory factors IL-1, IL-6, and TNF-α in the inflamed zebrafish in the comparative example were higher than in Examples 1-5, meaning that the anti-inflammatory effect of the comparative composition was not as good as that of the example composition.

[0110] This indicates that the formulations described in the examples were chosen based on synergistic considerations, rather than a simple combination of well-known anti-inflammatory foods. The compositions selected in Examples 1-5, together with bird's nest peptides, achieve a synergistic anti-inflammatory effect and have a better flavor, making them more palatable. Furthermore, the anti-inflammatory activity of the bird's nest peptide nanoparticles is stronger than that of the bird's nest peptide composition itself. This is because the nanoparticles have a larger specific surface area, and the cationic polyelectrolyte properties of chitosan can generate strong electrostatic interactions with mucus or negatively charged mucosal surfaces. This allows the nanoparticles to adhere to the small intestinal mucosa for a longer period, enhancing cell permeability and improving the bioavailability of the bird's nest peptide composition, thereby enhancing its anti-inflammatory activity.

[0111] This indicates that the bird's nest peptide composition exhibits a good synergistic anti-inflammatory effect, and the prepared nanoparticles and their stable Pickering emulsion further enhance the anti-inflammatory activity of the bird's nest peptide composition. Furthermore, the Pickering emulsion prepared with fish oil as the oil phase exhibits stronger anti-inflammatory activity than that prepared with corn oil as the oil phase. This is because fish oil is rich in EPA and DHA, which can increase the synthesis of leukotriene B5 and effectively reduce the levels of pro-inflammatory factors, thereby exerting an anti-inflammatory effect. In summary, the bird's nest peptide composition, bird's nest peptide nanoparticles, and their stable Pickering emulsion can control inflammation by reducing NO production and downregulating the expression of pro-inflammatory factors IL-1, IL-6, and TNF-α.

[0112] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention.

Claims

1. A Pickering emulsion, characterized in that, include: The dispersion and oil phase of nanoparticles; The mass ratio of the nanoparticle dispersion to the oil phase is (1~9):1, wherein the mass concentration of the nanoparticle dispersion is 0.1~5.0%. The nanoparticles include: Bird's nest peptide composition; Carrier, wherein the bird's nest peptide composition is loaded; The bird's nest peptide composition comprises, by weight, 0.1-3 parts bird's nest peptide, 10-20 parts green tea powder, 1-10 parts acerola cherry powder, 0.1-1 parts red algae extract, 0.1-1 parts Ganoderma lucidum extract, 0.1-1 parts Ginkgo biloba extract, and 0.1-1 parts licorice extract. The oil phase includes fish oil; The method for preparing the nanoparticles includes: (1) The solution containing the bird's nest peptide composition is added dropwise to the chitosan solution and stirred to obtain a mixture; (2) Add sodium tripolyphosphate solution dropwise to the mixture and stir to obtain the nanoparticles; The pH value of the chitosan solution is 4.0~5.

0.

2. The Pickering emulsion according to claim 1, characterized in that, The mass ratio of sodium tripolyphosphate to chitosan is 1:3 to 1:7, and the mass ratio of bird's nest peptide composition to chitosan is 1:3 to 1:

6.

3. The Pickering emulsion according to claim 1, characterized in that, The nanoparticles have a particle size of 150.5~201.0 nm.

4. The Pickering emulsion according to claim 1, characterized in that, The nanoparticles are provided in the form of a nanoparticle dispersion.

5. The Pickering emulsion according to claim 1, characterized in that, The concentration of the solution containing the bird's nest peptide composition is 1~10 mg / mL.

6. The Pickering emulsion according to claim 1, characterized in that, The concentration of the chitosan solution is 0.5~10 mg / mL, and the chitosan solution contains acetic acid.

7. The Pickering emulsion according to claim 1, characterized in that, The concentration of the sodium tripolyphosphate solution is 0.5~20 mg / mL.

8. The Pickering emulsion according to claim 1, characterized in that, In step (1), the stirring time is 20~40 min; In step (2), the stirring time is 40-50 min.

9. The Pickering emulsion according to claim 1, characterized in that, In step (2), the nanoparticle suspension obtained by stirring is centrifuged, washed and dried to obtain nanoparticle powder.

10. A method for preparing the Pickering emulsion according to any one of claims 1-9, characterized in that, include: The dispersion of the nanoparticles and the oil phase are mixed to obtain the Pickering emulsion.

11. The method according to claim 10, characterized in that, The method includes: The dispersion of the nanoparticles and the oil phase were sheared and stirred, and the resulting mixture was homogenized by microfluidic jet to obtain the Pickering emulsion.

12. The method according to claim 11, characterized in that, The shearing and stirring speed is 8000~12000 rpm, and the time is 1~5 min.

13. A food, medicine, or skincare product, characterized in that, It includes at least one of the following: the Pickering emulsion according to any one of claims 1-9.

14. The use of the Pickering emulsion according to any one of claims 1-9 in the preparation of pharmaceuticals or skin care products, characterized in that, The medicine or skin care product is used for anti-inflammatory purposes.

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