Protein powder for assisting in improving Alzheimer's disease and preparation method thereof

Abstract

The invention belongs to the technical field of food nutrition and health, and particularly discloses protein powder for assisting in improving Alzheimer's disease and a preparation method of the protein powder according to the concept of dietary therapy in China, and the protein powder is prepared from the following components in percentage by weight: 35-45% of egg white protein peptide powder and 55-65% of core function compound powder. According to the invention, egg white protein peptide is used as a high-quality matrix, and nine specific medicinal materials including hippocampus, egg yolk phospholipid, egg membrane peptide, walnut, fructus alpiniae oxyphyllae, gastrodia elata, mulberry, sargassum fusiforme, wolfberry, ginkgo leaf, fructus evodiae and dandelion are cooperatively compatible; aiming at seven core pathological links of Alzheimer's disease, such as A beta deposition, Tauopathy, neuroinflammation, oxidative stress, cholinergic deficiency, neurodystrophy and cerebral circulation disorder, a multi-target synergistic effect network is constructed, comprehensive neuroprotection is realized, and tests prove that the space learning and memory ability and the autonomous activity ability can be effectively improved.

Description

Technical Field

[0001] This invention belongs to the field of food nutrition and health technology, and relates to the prevention of neurodegenerative diseases, specifically to a protein powder that helps improve Alzheimer's disease and its preparation method. Background Technology

[0002] my country has a long tradition of prioritizing dietary supplements over medicinal supplements and prevention over drug treatment. Alzheimer's disease (AD) is the most common neurodegenerative disease, characterized by β-amyloid (Aβ) deposition, Tau protein hyperphosphorylation, neuroinflammation, oxidative stress, and neuronal loss. Currently, there is no cure, making prevention and early intervention crucial. Existing interventions involve medications (such as cholinesterase inhibitors and NMDA receptor antagonists) to alleviate symptoms, but their effectiveness is limited and they have significant side effects. Nutritional intervention is gaining increasing attention as a safe and sustainable prevention strategy.

[0003] Existing nutritional products for Alzheimer's disease prevention mostly focus on a single mechanism (such as antioxidation) or a few ingredients, lacking a systematic, multi-target formulation that addresses the multiple pathological mechanisms of Alzheimer's disease (Aβ toxicity, Tau lesions, inflammation, oxidative stress, cholinergic deficiency, and neurotrophic insufficiency). Furthermore, effectively integrating various components with significantly different properties (such as fat-soluble and water-soluble) into a stable, easy-to-use, and palatable protein powder presents a technological challenge.

[0004] Based on this, the present invention is proposed. Summary of the Invention

[0005] Based on the above reasons, one objective of this invention is to provide a protein powder that helps improve Alzheimer's disease. It uses egg white protein peptides as a high-quality protein base and scientifically combines various functional ingredients with neuroprotective, antioxidant, anti-inflammatory, brain circulation-improving, and key nutrient-providing properties. This allows the protein powder to have a multi-target synergistic effect and effectively prevent and help improve Alzheimer's disease. The second objective is to provide a method for preparing the above-mentioned protein powder, which achieves synergistic effects through specific process integration.

[0006] This invention is achieved through the following technical solution: A protein powder that helps improve Alzheimer's disease, by weight percentage, consists of the following components: 35-45% egg white protein peptide powder and 55-65% core functional compound powder; The core functional compound powder is composed of the following components by weight percentage: 6-11% seahorse ultrafine powder, 6-11% egg membrane peptide powder, 11-17% walnut ultrafine powder or walnut peptide powder, 3-7% Alpinia oxyphylla extract, 4-9% Gastrodia elata extract, 6-11% mulberry ultrafine powder or mulberry extract, 6-11% Sargassum fusiforme extract, 4-9% Lycium barbarum ultrafine powder or Lycium barbarum extract, 6-11% Ginkgo biloba extract, 1.5-5.5% Evodia rutaecarpa extract, 1.5-5.5% Taraxacum mongolicum extract, and the balance being egg yolk phospholipid powder.

[0007] Preferably, a protein powder that helps improve Alzheimer's disease comprises, by weight percentage: 40% egg white protein peptide powder and 60% core functional compound powder; The core functional compound powder is composed of the following components by weight percentage: 8.5% seahorse ultrafine powder, 8.5% egg membrane peptide powder, 13% walnut peptide powder, 5% Alpinia oxyphylla extract, 6.5% Gastrodia elata extract, 8.5% mulberry extract, 8.5% Sargassum fusiforme extract, 6.5% wolfberry extract, 8.5% ginkgo biloba extract, 3.5% Evodia rutaecarpa extract, 3.5% dandelion extract, and 19.5% egg yolk phospholipid powder.

[0008] Preferably, the ultrafine powder is obtained by ultrafine grinding of medicinal tissue, and its particle size D90≤50μm; the extract is a standardized extract powder of medicinal tissue.

[0009] The present invention also provides a method for preparing the above-mentioned protein powder, comprising the following steps: Raw material pretreatment: Prepare or obtain egg white protein peptide powder, seahorse ultrafine powder, egg yolk phospholipid powder, egg membrane peptide powder, walnut ultrafine powder or walnut peptide powder that meet the requirements, standardized extracts of Sargassum fusiforme, Alpinia oxyphylla, Gastrodia elata, Ginkgo biloba, Evodia rutaecarpa, and Taraxacum mongolicum, and standardized extracts or ultrafine powders of mulberry and wolfberry. Premixing: Under low humidity conditions, mix the ultrafine powder or standardized extract of the core functional ingredients in proportion to obtain the core functional compound powder. Overall mixing: Under low humidity conditions, the core functional compound powder and egg white protein peptide powder are mixed evenly in proportion to obtain a mixed powder; Post-processing: The mixed powder is homogenized, then sterilized at low temperature, and packaged with high barrier properties in a clean, low-humidity environment.

[0010] Preferably, the standardized extract is obtained by pretreatment, extraction, concentration, purification or refining, drying and standardization of medicinal tissue.

[0011] Preferably, the egg white protein peptide powder is prepared by enzymatic hydrolysis of fresh egg white liquid using a mixture of pepsin, alkaline protease, and papain, followed by concentration and drying, wherein small peptides with a molecular weight of less than 1000 Da account for ≥60%.

[0012] Preferably, the low humidity environment refers to an environment with a relative humidity of <40%; the mixing is carried out using a three-dimensional motion mixer or a double cone mixer.

[0013] Preferably, the homogenization process ensures that the particle size D90 of the final product is ≤75μm; the low-temperature sterilization is performed by cobalt-60 irradiation at a dose of 8-15kGy, or by low-temperature electron beam sterilization at a dose of 10-20kGy; and the high-barrier packaging is an aluminum foil composite film bag or packaging can filled with nitrogen or inert gas.

[0014] Compared with the prior art, the present invention has the following beneficial effects: 1. This invention uses egg white protein peptides as a high-quality matrix, and synergistically combines seahorse, egg yolk phospholipids, egg membrane peptides, and nine specific medicinal materials including walnut, Alpinia oxyphylla, Gastrodia elata, mulberry, Sargassum fusiforme, wolfberry, ginkgo leaf, Evodia rutaecarpa, and dandelion. Targeting the seven core pathological links of Alzheimer's disease—Aβ deposition, Tau lesions, neuroinflammation, oxidative stress, cholinergic deficiency, neurotrophic disorders, and cerebral circulatory disorders—it constructs a multi-target synergistic network to achieve comprehensive neuroprotection. Experiments have proven that it can effectively improve spatial learning and memory abilities and autonomous activity abilities.

[0015] 2. This invention employs ultra-fine grinding and standardized extraction to ensure the content and bioavailability of active ingredients. Three-dimensional mixing and homogenization ensure product uniformity and reconstitution properties. In particular, low-temperature sterilization and high-barrier packaging solve the problem of preserving the activity of heat-sensitive ingredients. Ultimately, the complex formula is transformed into a convenient, palatable, and highly stable daily protein powder that combines excellent efficacy with a good user experience. Detailed Implementation

[0016] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to specific embodiments. It should be understood that these descriptions are merely exemplary and not intended to limit the scope of the invention. Furthermore, descriptions of well-known structures and techniques are omitted in the following description to avoid unnecessarily obscuring the concept of the invention. Unless otherwise specified, specific conditions in the embodiments are performed under conventional conditions or conditions recommended by the manufacturer. Reagents or instruments used, unless otherwise specified, are all commercially available conventional products.

[0017] The first embodiment of the present invention provides a protein powder for assisting in the improvement of Alzheimer's disease, which, by weight percentage, consists of the following components: 35-55% egg white protein peptide powder and 45-65% core functional compound powder; the core functional compound powder consists of the following components by weight percentage: 6-11% seahorse ultrafine powder, 6-11% egg membrane peptide powder, 11-17% walnut ultrafine powder or walnut peptide powder, 3-7% Alpinia oxyphylla extract, 4-9% Gastrodia elata extract, 6-11% mulberry ultrafine powder or mulberry extract, 6-11% Sargassum fusiforme extract, 4-9% Lycium barbarum ultrafine powder or Lycium barbarum extract, 6-11% Ginkgo biloba extract, 1.5-5.5% Evodia rutaecarpa extract, 1.5-5.5% Taraxacum mongolicum extract, and the balance being egg yolk phospholipid powder.

[0018] In this embodiment, the formulation design of the present invention is based on a systematic and multi-target precise combination of complex multi-pathological mechanisms of Alzheimer's disease (AD), forming a powerful synergistic network: Targeting Aβ and Tau pathology, fucoxanthin from Sargassum fusiforme directly inhibits Aβ aggregation; the strong antioxidant capacity of mulberry, wolfberry, and walnut reduces oxidative stress, indirectly reducing the abnormal aggregation of Aβ and Tau; egg white protein peptides can also play a role by inhibiting the activity of related enzymes. The three synergistically block the core pathology of AD from different angles.

[0019] For neuroinflammation, Gastrodia elata and Taraxacum mongolicum, traditional anti-inflammatory herbs, are combined with the immunomodulatory effects of egg membrane peptides, Lycium barbarum polysaccharides, and Sargassum fusiforme polysaccharides to build a powerful anti-inflammatory team that jointly inhibits excessive activation of microglia and reduces neuroinflammation damage. Evodia rutaecarpa and other active ingredients can regulate neurotransmitters, helping to alleviate mood swings and anxiety, and reduce the stimulation of nerves by inflammation.

[0020] For cholinergic system deficiencies, egg yolk phospholipids directly provide choline, a key precursor for the synthesis of the neurotransmitter acetylcholine, thus treating the symptoms; the volatile oil components in Alpinia oxyphylla can regulate the cholinergic system, addressing the root cause. The two treat both the symptoms and the root cause, working synergistically to improve cognitive function.

[0021] For neurotrophic disorders and cerebral circulatory disorders, we select seahorse, a traditional nerve-nourishing medicinal material; walnut kernels rich in omega-3 fatty acids and egg membrane peptides to promote neuronal survival and repair; and ginkgo leaf extract to improve cerebral microcirculation, ensuring that nutrients and drugs can be delivered smoothly to the site of action.

[0022] To address oxidative stress, this invention utilizes a comprehensive free radical scavenging network comprised of mulberry (containing anthocyanins), wolfberry (containing wolfberry polysaccharides and betaine), walnut (containing vitamin E and polyphenols), ginkgo (containing flavonoid glycosides), Sargassum fusiforme (containing fucoxanthin), and egg white peptides. These different components work synergistically against different types of reactive oxygen species (ROS), resulting in an antioxidant efficiency far exceeding that of a single component.

[0023] As can be seen, this invention is based on the complex multi-pathological mechanisms of Alzheimer's disease and achieves comprehensive neuroprotection through multi-pathway synergy. Furthermore, this invention selects egg white protein peptides (EWP) as the matrix, which not only possesses activity itself, but also has the characteristics of easy absorption and low allergenicity, which can promote the absorption and bioavailability of other functional ingredients taken with it, thereby enhancing the overall effectiveness of the formulation.

[0024] In some preferred embodiments, the protein powder that helps improve Alzheimer's disease comprises, by weight percentage, the following components: 40% egg white protein peptide powder and 60% core functional compound powder; the core functional compound powder comprises, by weight percentage, the following components: 8.5% hippocampal ultrafine powder, 8.5% egg membrane peptide powder, 13% walnut peptide powder, 5% Alpinia oxyphylla extract, 6.5% Gastrodia elata extract, 8.5% mulberry extract, 8.5% Sargassum fusiforme extract, 6.5% Lycium barbarum extract, 8.5% Ginkgo biloba extract, 3.5% Evodia rutaecarpa extract, 3.5% Taraxacum mongolicum extract, and 19.5% egg yolk phospholipid powder.

[0025] In some specific embodiments, the ultrafine powder is obtained by ultrafine grinding of medicinal tissue, and its particle size D90≤50μm; the extract is a standardized extract powder of medicinal tissue.

[0026] In this embodiment, the standardized extracts are mainly standardized extracts of various Chinese herbal medicine components, wherein: The standardized extract of Alpinia oxyphylla contains nocaconone content of not less than 0.5% (w / w) and naringin content of not less than 0.8% (w / w); The total content of gastrodin and p-hydroxybenzyl alcohol in the standardized extract of Gastrodia elata is ≥1.0% (w / w), of which the content of gastrodin is ≥0.8% (w / w); The total anthocyanin content (as cyanidin-3-glucoside (C3G)) in the standardized mulberry extract is ≥5.0% (w / w), and the total polyphenol content in the mulberry ultrafine powder is ≥3.0% (w / w) and the total polysaccharide content is ≥20% (w / w). The standardized extract of wolfberry contains ≥40.0% (w / w) wolfberry polysaccharides; The standardized extract of Sargassum fusiforme contains ≥2.0% (w / w) fucoxanthin. The standardized extract of Ginkgo biloba leaves contained 24% flavonoid glycosides and 6% (w / w) terpene lactones. The total content of evodiamine+ in the standardized extract of Evodia rutaecarpa is ≥3.0% (w / w); The standardized dandelion extract contains ≥2.0% (w / w) chlorogenic acid and ≥8.0% (w / w) total flavonoids. The second embodiment of the present invention provides a method for preparing the above-mentioned protein powder, comprising the following steps: Raw material pretreatment: Prepare or obtain egg white protein peptide powder, seahorse ultrafine powder, egg yolk phospholipid powder, egg membrane peptide powder, walnut ultrafine powder or walnut peptide powder that meet the requirements, standardized extracts of Sargassum fusiforme, Alpinia oxyphylla, Gastrodia elata, Ginkgo biloba, Evodia rutaecarpa, and Taraxacum mongolicum, and standardized extracts or ultrafine powders of mulberry and wolfberry. Premixing: Under low humidity conditions, mix the ultrafine powder or standardized extract of the core functional ingredients in proportion to obtain the core functional compound powder. Overall mixing: Under low humidity conditions, the core functional compound powder and egg white protein peptide powder are mixed evenly in proportion to obtain a mixed powder; Post-processing: The mixed powder is homogenized, then sterilized at low temperature, and packaged with high barrier properties in a clean, low-humidity environment.

[0027] In some specific embodiments, the egg white protein peptide powder is prepared from fresh egg white liquid by enzymatic hydrolysis with a mixture of pepsin, alkaline protease, and papain, followed by concentration and drying. The proportion of small peptides with a molecular weight less than 1000 Da is ≥60%. This invention decomposes large-molecule egg white liquid into absorbable small-molecule protein peptides through mixed enzymatic hydrolysis. Testing has shown that these peptides exhibit good antioxidant activity and can scavenge O2· - The effective half-maximum inhibitory concentration (IC50) for ·OH groups is 0.5%. 50 The half-maximal inhibitory concentrations (IC50) for DNA damage were 0.11 mg / mL and 0.16 mg / mL, respectively. 50 It is 0.89 mg / mL.

[0028] In some specific embodiments, the standardized extract is obtained by pretreatment, extraction, concentration, purification or refining, drying and standardization of medicinal tissue.

[0029] In this embodiment, powders that meet the requirements are first prepared based on the physicochemical properties of the ingredients in the formula. The main methods for preparing the powders are ultrafine grinding and standardized extraction technology. For edible medicinal materials such as seahorse, mulberry, and wolfberry, ultrafine grinding after low-temperature drying can be used to destroy the cell wall structure of the raw materials, greatly increasing their specific surface area, making the internal active ingredients easier to dissolve and be absorbed by the human body, and significantly improving bioavailability. For key Chinese medicines such as ginkgo leaves, gastrodia elata, and evodia rutaecarpa, standardized extract powders are used to ensure the accuracy, stability, and controllability of the content of the main active ingredients in each batch of products, thereby ensuring the consistency and reliability of product efficacy.

[0030] The preparation process of standardized extracts typically follows a core pathway: pretreatment → extraction → concentration → purification / refining → drying → standardization. Common extraction methods include solvent extraction (water extraction, alcohol extraction, and water extraction with alcohol precipitation) and supercritical fluid extraction. Reduced pressure concentration and vacuum concentration can prevent the destruction of heat-sensitive components in the medicinal material. Purification and refining are key steps in achieving standardization, with common methods including macroporous adsorption resin chromatography and membrane separation. Drying methods are selected based on the characteristics of the medicinal material, typically using spray drying, vacuum belt drying, and freeze drying. This invention takes a standardized extract of ginkgo leaves as an example. The preparation process is as follows: dried ginkgo leaves are taken, pulverized, and extracted with 60% ethanol under dynamic hot reflux. After filtration, the extract is concentrated under reduced pressure to recover the ethanol and obtain a concentrated solution. The concentrated solution is purified using an AB-8 macroporous adsorption resin column. Impurities are first washed away with water, and then eluted with a gradient of 70% ethanol solution. The eluent rich in flavonoid glycosides and terpene lactones is collected. The eluent is concentrated by nanofiltration membrane and then sent to a spray drying tower for drying to obtain a brownish-yellow powder. Finally, the extract is standardized to contain 24.0% total flavonoid glycosides, 6.0% total terpene lactones, and ginkgolic acid content not exceeding 5 ppm.

[0031] Standardized extraction of medicinal materials is a routine procedure in this field, and its preparation process will not be described in detail here. Alternatively, standardized extracts of medicinal materials that meet the requirements can be purchased directly.

[0032] In some specific embodiments, the low humidity environment refers to an environment with a relative humidity of <40%; the mixing is carried out using a three-dimensional motion mixer or a double cone mixer; the homogenization treatment ensures that the particle size D90 of the final product is ≤75μm; the low-temperature sterilization is carried out using cobalt-60 irradiation at a dose of 8-15kGy, or low-temperature electron beam sterilization at a dose of 10-20kGy; and the high-barrier packaging is an aluminum foil composite film bag or packaging can filled with nitrogen or inert gas.

[0033] This invention utilizes a three-dimensional motion mixer or a double-cone mixer for multiple mixing processes in a low-humidity environment. This effectively solves the problems of uneven mixing and clumping caused by significant differences in the density, particle size, and electrostatic properties of the components, ensuring that each scoop contains a precise proportion of each ingredient and accurate dosage. The final product undergoes homogenization to achieve a uniform and fine powder particle size, greatly improving the product's reconstitution and mouthfeel. It is smooth and free of gritty texture, resolving palatability issues and increasing user compliance for long-term use. Low-temperature sterilization replaces traditional heat sterilization, effectively killing microorganisms and ensuring product safety while avoiding the degradation temperatures of heat-sensitive components. This results in a protein powder bioactivity retention rate of over 95%, effectively reducing the loss of functional components. Finally, high-barrier packaging effectively isolates oxygen and moisture, preventing oxidative rancidity of oils and the slow oxidation of components such as polyphenols. This ensures the product's flavor and functional activity stability throughout its shelf life, extending its shelf life. Therefore, the preparation method of this invention maximizes the preservation of the activity of each functional component and ensures the product's uniformity, stability, and safety.

[0034] The following will disclose specific embodiments for implementing this application, along with corresponding comparative examples to demonstrate the relevant technical effects of this application.

[0035] Example 1: Preparation of protein powder Egg white protein peptide powder: Made from fresh egg white liquid, with the addition of pepsin, alkaline protease and papain for complex enzymatic hydrolysis, and then concentrated and dried to obtain high-quality egg white protein peptide powder with a molecular weight of less than 1000 Da and a small peptide content of more than 60%. Seahorse ultrafine powder: Selected seahorses are cleaned, freeze-dried or low-temperature dried, and then ultrafine pulverized to a particle size D90≤50μm; Egg yolk phospholipid powder: High-purity (≥70% PC) powdered egg yolk phospholipid is selected; Egg membrane peptide powder: Egg membrane peptide powder is prepared by washing and sterilizing eggs, separating the shell membrane by water method, and performing secondary enzymatic hydrolysis with neutral protease and alkaline protease. Walnut peptide powder: Walnut kernels are deastringentized, low-temperature roasted, enzymatically hydrolyzed, separated and purified, and then freeze-dried to make walnut peptide powder; Standardized extracts of Alpinia oxyphylla, Gastrodia elata, Morus alba, Sargassum fusiforme, Lycium barbarum, Ginkgo biloba, Evodia rutaecarpa and Taraxacum mongolicum: Select medicinal materials, use supercritical CO2 extraction process, collect the extract, vacuum concentrate and freeze dry to obtain extract powder, and then standardize to obtain standardized extract powder. Mixing: Under low humidity conditions (ambient humidity 38%), by weight percentage, first mix 8.5% seahorse ultrafine powder, 8.5% egg membrane peptide powder, 13% walnut peptide powder, 5% Alpinia oxyphylla extract, 6.5% Gastrodia elata extract, 8.5% mulberry extract, 8.5% Sargassum fusiforme extract, 6.5% wolfberry extract, 8.5% Ginkgo biloba extract, 3.5% Evodia rutaecarpa extract, 3.5% dandelion extract, and 19.5% egg yolk phospholipid powder evenly to obtain the core functional compound powder; by weight percentage, take 60% of the core functional compound powder and 40% egg white protein peptide powder, and continue to mix evenly to obtain the mixed powder; Post-processing: The mixed powder was homogenized to a particle size D90≤75μm using a three-dimensional motion mixer, and then sterilized by cobalt-60 irradiation at a dose of 12kGy. The powder was then packaged in nitrogen-filled aluminum foil composite film bags in a low-humidity clean environment to obtain the finished protein powder.

[0036] Example 2 Preparation of protein powder Egg white protein peptide powder: Made from fresh egg white liquid, with the addition of pepsin, alkaline protease and papain for complex enzymatic hydrolysis, and then concentrated and dried to obtain high-quality egg white protein peptide powder with a molecular weight of less than 1000 Da and a small peptide content of more than 60%. Seahorse, mulberry, and goji berry ultrafine powder: Selected seahorse, mulberry, and goji berries are washed, dried at low temperature, and then pulverized to a particle size D90≤50μm using ultrafine pulverization technology; Egg yolk phospholipid powder: Liquid egg yolk phospholipids are processed into powder using low-temperature spray drying technology; Egg membrane peptide powder: Egg membrane peptide powder is prepared by washing and sterilizing eggs, separating the shell membrane by water method, and performing secondary enzymatic hydrolysis of the egg membrane with neutral protease and alkaline protease. Walnut powder: Walnut kernels are processed into walnut powder through de-astringency removal, low-temperature baking, and ultra-fine grinding; Standardized extracts of Alpinia oxyphylla, Gastrodia elata, Sargassum fusiforme, Ginkgo biloba, Evodia rutaecarpa and Taraxacum mongolicum: The medicinal materials were extracted by ethanol reflux, concentrated under reduced pressure and spray dried to obtain extract powder, which was then standardized to obtain standardized extract powder. Mixing: Under low humidity conditions (ambient humidity 35%), by weight percentage, first mix 11% seahorse ultrafine powder, 6% egg membrane peptide powder, 17% walnut powder, 7% Alpinia oxyphylla extract, 9% Gastrodia elata extract, 11% mulberry ultrafine powder, 11% Sargassum fusiforme extract, 9% Lycium barbarum ultrafine powder, 6% Ginkgo biloba extract, 1.5% Evodia rutaecarpa extract, 1.5% Taraxacum mongolicum extract, and 10% egg yolk phospholipid powder evenly to obtain the core functional compound powder; by weight percentage, take 62% of the core functional compound powder and 38% egg white protein peptide powder, and continue to mix evenly to obtain the mixed powder; Post-processing: The mixed powder was homogenized to a particle size D90≤75μm using a double cone mixer, and then sterilized by low-temperature electron beam at a dose of 15kGy. The powder was then packaged in nitrogen-filled aluminum foil cans in a low-humidity clean environment to obtain the finished protein powder.

[0037] Example 3 Preparation of protein powder The raw material pretreatment method is the same as in Example 1; Mixing: Under low humidity conditions (ambient humidity 38%), by weight percentage, first mix 11% seahorse ultrafine powder, 6% egg membrane peptide powder, 11% walnut peptide powder, 7% Alpinia oxyphylla extract, 9% Gastrodia elata extract, 6% mulberry extract, 6% Sargassum fusiforme extract, 4% Lycium barbarum extract, 11% Ginkgo biloba extract, 1.5% Evodia rutaecarpa extract, 1.5% Taraxacum mongolicum extract, and 26% egg yolk phospholipid powder evenly to obtain the core functional compound powder; by weight percentage, take 55% of the core functional compound powder and 45% egg white protein peptide powder, and continue to mix evenly to obtain the mixed powder; Post-processing: The mixed powder was homogenized to a particle size D90≤75μm using a three-dimensional motion mixer, and then sterilized by cobalt-60 irradiation at a dose of 8kGy. The powder was then packaged in nitrogen-filled aluminum foil composite film bags in a low-humidity clean environment to obtain the finished protein powder.

[0038] Example 4: Preparation of protein powder The raw material processing method is the same as in Example 1; Mixing: Under low humidity conditions (ambient humidity 38%), by weight percentage, first mix 6% seahorse ultrafine powder, 11% egg membrane peptide powder, 17% walnut peptide powder, 3% Alpinia oxyphylla extract, 4% Gastrodia elata extract, 11% mulberry extract, 11% Sargassum fusiforme extract, 9% Lycium barbarum extract, 6% Ginkgo biloba extract, 3.5% Evodia rutaecarpa extract, 5.5% Taraxacum mongolicum extract, and 13% egg yolk phospholipid powder evenly to obtain the core functional compound powder; by weight percentage, take 61% of the core functional compound powder and 39% egg white protein peptide powder, and continue to mix evenly to obtain the mixed powder; Post-processing: The mixed powder was homogenized to a particle size D90≤75μm using a three-dimensional motion mixer, and then sterilized by low-temperature electron beam at a dose of 20kGy. The powder was then packaged in nitrogen-filled aluminum foil composite film bags in a low-humidity clean environment to obtain the finished protein powder.

[0039] Example 5 Preparation of protein powder The raw material processing method is the same as in Example 1; Mixing: Under low humidity conditions (ambient humidity 35%), by weight percentage, first mix 11% seahorse ultrafine powder, 8.5% egg membrane peptide powder, 11% walnut peptide powder, 5% Alpinia oxyphylla extract, 6.5% Gastrodia elata extract, 6% mulberry extract, 11% Sargassum fusiforme extract, 4% Lycium barbarum extract, 8.5% Ginkgo biloba extract, 5.5% Evodia rutaecarpa extract, 5.5% Taraxacum mongolicum extract, and 17.5% egg yolk phospholipid powder evenly to obtain the core functional compound powder; then, by weight percentage, take 65% of the core functional compound powder and 35% egg white protein peptide powder, and continue to mix evenly to obtain the mixed powder; Post-processing: The mixed powder was homogenized to a particle size D90≤75μm using a three-dimensional motion mixer, and then sterilized by low-temperature electron beam at a dose of 10kGy. The powder was then packaged in nitrogen-filled aluminum foil composite film bags in a low-humidity clean environment to obtain the finished protein powder.

[0040] Comparative Example 1: Preparation of Protein Powder The preparation process is exactly the same as in Example 1, except that the addition of Sargassum fusiforme extract is omitted and replaced with an equal amount of egg white protein peptide powder. The proportions of other components are the same as in Example 1.

[0041] Comparative Example 2: Preparation of Protein Powder The preparation process and component ratios are exactly the same as in Example 1, except that the homogenization step is omitted.

[0042] Comparative Example 3: Preparation of Protein Powder The preparation process and component ratios are exactly the same as in Example 1, except that low-temperature electron beam sterilization is replaced with traditional high-temperature instantaneous sterilization (UHT).

[0043] Comparative Example 4: Preparation of Protein Powder The preparation process and component ratio are exactly the same as in Example 1, except that the nitrogen-filled aluminum foil composite film packaging is replaced with ordinary polyethylene (PE) plastic bag packaging.

[0044] Experimental Example 1: Detection of Physicochemical Indicators Using Example 1 and Comparative Examples 1-4 as samples, the moisture content, particle size distribution, phosphatidylcholine (PC) retention rate, total flavonoid retention rate and fucoxanthin retention rate were detected immediately after preparation.

[0045] Moisture content was determined using the direct drying method, Method 1, of GB 5009.3-2016 "National Food Safety Standard - Determination of Moisture in Food". Particle size distribution was determined using the laser diffraction method, GB / T 19077-2016 "Particle Size Analysis".

[0046] Detection of phosphatidylcholine retention: A suitable amount of sample was weighed and extracted with chloroform:methanol = 2:1 (v / v). After centrifugation, the lower organic phase was collected and concentrated by nitrogen blowing. High-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) was used for detection. The chromatographic column was a silica gel column (4.6 × 250 mm, 5 μm). The mobile phase was A: n-hexane:isopropanol:ethyl acetate = 8:8:1 (v / v), containing 0.08% triethylamine; B: acetic acid. The flow rate was 1.0 mL / min, and the column temperature was 40℃. The ELSD parameters were: drift tube temperature 80℃, carrier gas (N2) flow rate 2.0 L / min. The phosphatidylcholine content was calculated using the external standard method, and then the retention rate was calculated, with the highest content defined as 100%.

[0047] Determination of total flavonoid retention rate: Weigh an appropriate amount of sample, extract with 60% ethanol using ultrasound, and bring to a final volume; take an appropriate amount of extract, add 5% sodium nitrite solution, shake well, and let stand for 6 minutes; add 10% aluminum nitrate solution, shake well, and let stand for 6 minutes; add 4% sodium hydroxide solution, bring to a final volume with 60% ethanol, shake well, and let stand for 15 minutes, then measure the absorbance (A) at a wavelength of 510 nm; prepare a standard curve using rutin as a standard, calculate the total flavonoid content (calculated as rutin) in the sample according to the standard curve equation, and then calculate the retention rate, with the highest content being 100%.

[0048] Determination of fucoxanthin retention: Accurately weigh the sample, add an appropriate amount of ethanol, extract by ultrasonication in the dark, centrifuge, collect the supernatant, and filter through a membrane for analysis. High-performance liquid chromatography (HPLC) was used for detection. The chromatographic column was a C18 column (4.6 × 250 mm, 5 μm). Mobile phase A: methanol; B: water (gradient elution, 80% A → 100% A in 20 min), flow rate 1.0 mL / min, column temperature 30℃, and detection wavelength 450 nm. A standard curve was prepared using fucoxanthin standards. The content was calculated using the external standard method, and then the retention rate was calculated, with the highest content defined as 100%.

[0049] The test results are as follows: Table 1. Results of Physicochemical Indicators Tests .

[0050] As shown in Table 1, compared with Example 1, Comparative Example 1 omitted the addition of Sargassum fusiforme extract, and fucoxanthin was not detected, while other indicators were not significantly different; Comparative Example 2 omitted the homogenization step, resulting in a significant increase in particle size D90, poorer flowability, and a significantly higher moisture content due to mixing at high humidity, which is not conducive to product stability; Comparative Example 3 used high-temperature sterilization, which led to a large amount of degradation of heat-sensitive components, and a sharp decrease in the retention rates of phosphatidylcholine and total flavonoids, which were only 62% and 75%, respectively. Fucoxanthin, due to its certain thermal stability, was lost less, but it also decreased significantly, indicating that low-temperature sterilization is crucial for protecting active ingredients; Comparative Example 4 used ordinary packaging, which did not affect the initial content of the sample.

[0051] Test Example 2 Accelerated Stability Test Example 1 and Comparative Example 4 were placed in a constant temperature and humidity chamber at 40±2℃ and 75±5% relative humidity for accelerated testing. Samples were taken at 30 and 60 days to detect changes in peroxide value, acid value, phosphatidylcholine retention rate, total flavonoid retention rate, and color and flavor.

[0052] The peroxide value was determined using the first titration method of GB 5009.227-2016 "National Food Safety Standard - Determination of Peroxide Value in Food"; the acid value was determined using GB 5009.229-2016 "National Food Safety Standard - Determination of Acid Value in Food".

[0053] The test results are as follows: Table 2 Accelerated Test Results .

[0054] As shown in Table 2, the peroxide value and acid value of Comparative Example 4 increased significantly over time compared to Example 1. After 60 days, the peroxide value and acid value of Comparative Example 4 changed significantly, indicating severe oxidative rancidity. Under accelerated conditions of high temperature and high humidity, the degradation rate of the active ingredients (phosphatidylcholine and total flavonoids) in Comparative Example 4 was significantly faster than that in Example 1. Over time, Comparative Example 4 showed obvious darkening of color and an unpleasant rancid odor, while Example 1 remained relatively stable. It can be seen that high-barrier nitrogen-filled packaging can effectively delay oxidation and protect the stability of product flavor and functional components throughout the shelf life, which is a key technology for ensuring product shelf life.

[0055] Experimental Example 3: Detection of In Vitro Activity Indicators of Protein Powder Oxygen radical scavenging capacity (ORAC) measures a sample's ability to neutralize peroxyl radicals (ROO•). It is highly correlated with oxidative stress in vivo and can comprehensively reflect the synergistic effects of multiple antioxidant components (polyphenols, peptides, polysaccharides, etc.) in a product. Using fluorescein, peroxyl radicals were generated by adding AAPH (2,2'-Azobis(2-amidinopropane)dihydrochloride) to phosphate buffer at pH 7.4 at 37°C, and sample extracts of different concentrations were added. Fluorescence decay curves were monitored. Results are expressed as Trolox (a water-soluble vitamin E analog), i.e., μmol TE / g powder.

[0056] Anti-Aβ aggregation activity assay: The thioflavin T fluorescence assay was used. ThT is a fluorescent dye that significantly enhances fluorescence intensity after binding to Aβ fibers. The fluorescence intensity is directly proportional to the Aβ fiber content. 42Monomers and sample extracts of different concentrations were co-incubated at 37°C for 48 hours. Samples were taken at 2-hour intervals, ThT solution was added, and fluorescence intensity was immediately detected. Fluorescence intensity-time curves were plotted to observe whether the samples delayed fiber formation or reduced the final fiber yield. The inhibition rate (%) of the samples on Aβ fiber formation was calculated after 24 hours.

[0057] Acetylcholinesterase inhibitory activity assay: Acetylcholinesterase (AChE) hydrolyzes the substrate acetylthiocholine (ATCI) to generate thiocholine, which reacts with DTNB (5,5'-dithiobis(2-nitrobenzoic acid)) to produce a yellow product with a characteristic absorption peak at 412 nm. Inhibitors reduce the reaction rate. Different concentrations of sample, AChE enzyme solution, and DTNB were added to 96-well plates. After incubation, the substrate ATCI was added, and the rate of absorbance change was immediately monitored at 412 nm. The IC50 value of the sample to AChE activity was calculated. 50 value.

[0058] Anti-neuroinflammatory activity assay: Overactivation of microglia (immune cells in the brain) is the core of AD neuroinflammation. An inflammation model was established by stimulating microglia (BV2 cell line) with lipopolysaccharide (LPS). The safe concentration range of the sample for BV2 cells was determined. BV2 cells were pretreated with the sample within the safe concentration for 1-2 hours, followed by LPS stimulation for 24 hours. Cell supernatant was collected, and nitric oxide (NO) content was detected by Griess reagent method to calculate the inhibition rate.

[0059] Neuroprotective activity assay: Human neuroblastoma cells (SH-SY5Y) were selected, pretreated with a safe concentration of sample for a period of time, and then Aβ was added. 25-35 Oligomers or Aβ 42 After co-incubation for 24-48 hours, cell viability was detected using a cytotoxicity assay.

[0060] The protein powders prepared in Examples 1-5 and Comparative Example 1 were subjected to in vitro activity testing. All samples were tested using a uniform method to prepare aqueous extracts. The test results are as follows: Table 3 Results of in vitro bioactivity assay .

[0061] As can be seen from the experimental results in Table 3, Examples 1-5 all showed significant effects on five in vitro activity indicators, fully demonstrating that the formulation of this invention can achieve the core function of preventing AD under different ratios. Example 1 showed stable and excellent performance on all indicators without any obvious shortcomings, verifying its comprehensiveness and reliability as a basic formulation; Example 2 replaced part of the standard extract with ultrafine powder, and although the indicators were not optimal, it still maintained good basic activity, and its cost may be more advantageous; Example 3 strengthened the nootropic components such as egg yolk phospholipids (choline source), seahorse, and gastrodia, which showed the best performance in inhibiting acetylcholinesterase, proving that it has the strongest ability to improve the function of the cholinergic system; Example 4 increased the proportion of antioxidant components such as mulberry, wolfberry, and Sargassum fusiforme, which had the highest oxygen free radical absorption capacity value, and its anti-neuroinflammatory (NO inhibition) effect was also the most prominent, which was highly consistent with the formulation design target; Example 5 specifically increased the amount of seahorse and Sargassum fusiforme, which showed the strongest effect in inhibiting Aβ aggregation, highlighting its unique advantage in targeting Aβ pathology. Although the five sets of formulations provided by this invention each have their own focus, they can all effectively cover multiple core pathological aspects of Alzheimer's disease (AD), and their in vitro activity characteristics are highly consistent with the original design intent of the formulations. Compared with Example 1, Comparative Example 1 lacks Sargassum fucoxanthin, and its ability to inhibit Aβ aggregation decreases sharply, significantly worse than all other examples. This proves that the formulation of this invention is an organic whole, with each component being indispensable and exhibiting a significant synergistic effect.

[0062] Experimental Example 4: Animal Behavior of the AD Model Experimental Methods: Sixty 3-month-old 3xTg-AD transgenic AD model mice were randomly divided into two groups (n=30 / group): the AD model group and the product intervention group. Wild-type C57BL / 6J mice with the same background were used as a blank control group (n=30 / group). All mice were housed in an SPF-grade animal facility with free access to water and food, and underwent a 12 / 12-hour light / dark cycle. The experimental procedure was reviewed and approved by the institution's animal ethics committee.

[0063] The intervention group received the protein powder from Example 1 of this invention orally daily at a dose of 500 mg / kg bw (calculated based on body weight) in addition to a normal diet. The AD model group and the blank control group received the same amount of normal diet. The intervention lasted for 6 months, during which the animals were in good condition and there were no deaths. Behavioral tests were performed on the mice at 9 months of age, in the following order: first, the open field test (to assess basic activity and anxiety), followed by the Morris water maze test (to assess spatial learning and memory abilities). Drug administration was suspended during the testing period.

[0064] Open field test: Mice were placed in the center of an open field box measuring 50cm × 50cm × 40cm, and spontaneous activity was recorded for 10 minutes. The main observation indicator was the total distance traveled (cm), and the results are shown in Table 4 below. Analysis showed that the total distance traveled in the product intervention group was significantly higher than that in the AD model group (p < 0.001), and completely recovered to a level comparable to the blank control group (p > 0.05). This result indicates that the product of this invention effectively improved the sluggishness and lethargy in AD model mice, significantly enhancing their activity level and willingness to explore.

[0065] Table 4 Record of Open Field Test Results .

[0066] Morris Water Maze Test: A 5-day experiment was conducted. The first 4 days were for a navigational test, with 4 tests per day. The escape latency of mice finding the hidden platform was recorded, and the results are shown in Table 5 below. Analysis showed that from day 3 onwards, the escape latency of the product intervention group was significantly shorter than that of the AD model group (p<0.05), and there was no significant difference compared with the blank control group (p>0.05). The results indicate that the product of this invention can significantly improve the spatial learning ability of AD model mice.

[0067] Table 5 Record of Positioning Navigation Test Results .

[0068] On day 5, the platform was removed, and a 60-second spatial exploration experiment was conducted. The number of times the mice traversed the original platform location was recorded, and the results are shown in Table 6 below. Analysis showed that the number of traversals in the product intervention group was significantly higher than that in the AD model group (p<0.001), and there was no statistically significant difference compared to the blank control group (p>0.05). This result confirms that the product intervention of this invention restored 37.8% of the spatial memory ability of AD model mice, demonstrating a significant effect.

[0069] Table 6 Record of Space Exploration Experiment Results .

[0070] Animal behavioral experiments have demonstrated that long-term administration of the protein powder of this invention can significantly improve the spatial learning and memory abilities and autonomous activity abilities of 3xTg-AD model mice. The effects are reflected in a shortened escape latency in the water maze, a 37.8% increase in the number of times mice traverse the original platform, and a 61.0% increase in the total distance traveled in the open field experiment. These significant improvements in behavioral indicators, combined with the absence of lethargy in the mice, strongly support the application prospects of this invention's product in the prevention and adjunctive treatment of Alzheimer's disease.

[0071] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Any simple modifications or equivalent changes made to the above embodiments based on the technical essence of the present invention shall fall within the protection scope of the present invention.

Claims

1. A protein powder that helps improve Alzheimer's disease, characterized in that, By weight percentage, it consists of the following components: 35-45% egg white protein peptide powder and 55-65% core functional compound powder; The core functional compound powder is composed of the following components by weight percentage: 6-11% seahorse ultrafine powder, 6-11% egg membrane peptide powder, 11-17% walnut ultrafine powder or walnut peptide powder, 3-7% Alpinia oxyphylla extract, 4-9% Gastrodia elata extract, 6-11% mulberry ultrafine powder or mulberry extract, 6-11% Sargassum fusiforme extract, 4-9% Lycium barbarum ultrafine powder or Lycium barbarum extract, 6-11% Ginkgo biloba extract, 1.5-5.5% Evodia rutaecarpa extract, 1.5-5.5% Taraxacum mongolicum extract, and the balance being egg yolk phospholipid powder.

2. The protein powder according to claim 1, characterized in that, By weight percentage, it consists of the following components: 40% egg white protein peptide powder and 60% core functional compound powder; The core functional compound powder is composed of the following components by weight percentage: 8.5% seahorse ultrafine powder, 8.5% egg membrane peptide powder, 13% walnut peptide powder, 5% Alpinia oxyphylla extract, 6.5% Gastrodia elata extract, 8.5% mulberry extract, 8.5% Sargassum fusiforme extract, 6.5% wolfberry extract, 8.5% ginkgo biloba extract, 3.5% Evodia rutaecarpa extract, 3.5% dandelion extract, and 19.5% egg yolk phospholipid powder.

3. The protein powder according to claim 1, characterized in that, The ultrafine powder is obtained by ultrafine grinding of medicinal tissue, and its particle size D90≤50μm; the extract is a standardized extract powder of medicinal tissue.

4. The method for preparing protein powder according to any one of claims 1-3, characterized in that, Includes the following steps: Raw material pretreatment: Prepare or obtain egg white protein peptide powder, seahorse ultrafine powder, egg yolk phospholipid powder, egg membrane peptide powder, walnut ultrafine powder or walnut peptide powder that meet the requirements, standardized extracts of Sargassum fusiforme, Alpinia oxyphylla, Gastrodia elata, Ginkgo biloba, Evodia rutaecarpa, and Taraxacum mongolicum, and standardized extracts or ultrafine powders of mulberry and wolfberry. Premixing: Under low humidity conditions, mix the ultrafine powder or standardized extract of the core functional ingredients in proportion to obtain the core functional compound powder. Overall mixing: Under low humidity conditions, the core functional compound powder and egg white protein peptide powder are mixed evenly in proportion to obtain a mixed powder; Post-processing: The mixed powder is homogenized, then sterilized at low temperature, and packaged with high barrier properties in a clean, low-humidity environment.

5. The preparation method according to claim 4, characterized in that, The standardized extract is obtained by pretreatment, extraction, concentration, purification or refining, drying and standardization of medicinal tissues.

6. The preparation method according to claim 4, characterized in that, The egg white protein peptide powder is prepared by enzymatic hydrolysis of fresh egg white liquid using a mixture of pepsin, alkaline protease, and papain, followed by concentration and drying. The proportion of small peptides with a molecular weight of less than 1000 Da is ≥60%.

7. The preparation method according to claim 4, characterized in that, The low humidity environment refers to an environment with a relative humidity of <40%; the mixing is carried out using a three-dimensional motion mixer or a double cone mixer.

8. The preparation method according to claim 4, characterized in that, The homogenization process ensures that the particle size D90 of the final product is ≤75μm; the low-temperature sterilization is performed by cobalt-60 irradiation at a dose of 8-15kGy, or by low-temperature electron beam sterilization at a dose of 10-20kGy; the high-barrier packaging is an aluminum foil composite film bag or packaging can filled with nitrogen or inert gas.