Application of casein phosphopeptides to regulation of body tryptophan metabolism

Abstract

The invention belongs to the technical field of food, and relates to an application of casein phosphopeptides in regulating tryptophan metabolism of an organism. Research results show that the casein phosphopeptides are obtained through a serotonin metabolic pathway and / or a kynurenine metabolic pathway; the content of tryptophan in a body is increased, and / or the content of at least one metabolite (5-methoxyindole-3-acetic acid, N-acetyl serotonin, kynurenine, 2-picolinic acid and indole-3-formaldehyde) in a tryptophan metabolic pathway is increased, so that the effects of emotion regulation, sleep regulation and / or neuroprotection are favorably exerted. Besides, the invention has no special requirements on the specific application mode of the casein phosphopeptides and the form of food containing the casein phosphopeptides, has convenience in implementation, and also provides a new thought for the application research of the casein phosphopeptides.

Description

Technical Field

[0001] This invention belongs to the food field and relates to the use of a casein phosphopeptide to help regulate tryptophan metabolism in the body. Background Technology

[0002] Casein phosphopeptides (CPP) are bioactive polypeptides produced using biotechnology from cow's milk or casein. Since their discovery, they have received widespread attention in the food and health supplement industries. Numerous studies have confirmed that CPP can effectively promote the absorption and utilization of divalent mineral nutrients such as calcium, iron, and zinc in the human body. In addition, CPP also has various physiological functions such as anti-caries and blood pressure regulation.

[0003] Regarding the prevention of tooth decay, traditional views hold that chewing cheese after meals stimulates saliva production, and the alkaline buffering effect of saliva neutralizes the acidic substances in dental plaque, thereby reducing erosion of tooth enamel. Recent studies have found that CPP contained in cheese can bind calcium ions from food to the site of decay, inhibiting the enamel demineralization process and thus playing a role in preventing tooth decay.

[0004] Regarding blood pressure regulation, studies have shown that during fermentation in the milk matrix, *Lactobacillus helveticus* releases specific peptides from milk proteins through proteolytic hydrolysis. These peptides possess inhibitory activity against angiotensin-converting enzyme (ACE) or opioid peptide-like effects. CPP may play a synergistic role in this process, enhancing its antihypertensive effect in vivo.

[0005] Currently, CPP is widely used in a variety of products. In the field of nutrition and health foods, CPP is often used in fortified calcium, iron, and zinc supplements, dairy products (such as milk powder, school formula milk, high-calcium low-fat milk, etc.), children's foods (such as infant rice cereal, high-calcium biscuits, etc.), soy products (such as high-calcium soy milk powder, calcium-fortified tofu, etc.), as well as nutritional cereals, chewing gum, and other products. Its addition not only improves product formulations and enhances the bioavailability of minerals, but also promotes the upgrading and transformation of related products.

[0006] In the field of oral care, CPP has also been given new applications. For example, some studies have combined CPP with strontium ions to enhance the efficacy of oral care products. CPP can promote the adsorption and delivery of strontium ions on the tooth surface, synergistically enhancing their effects in resisting dentin hypersensitivity, promoting tooth mineralization, and preventing tooth erosion.

[0007] The application areas of casein phosphopeptides are constantly expanding in existing technologies. For example, reference 1, through a mouse model study, revealed that a combination of casein phosphopeptides and Bifidobacteria can work synergistically to significantly enhance immunity and improve the body's antioxidant capacity. This research data further confirms that even casein phosphopeptide monomers possess good immunomodulatory and antioxidant activities, opening up new avenues for their application in functional foods and health products.

[0008] References:

[0009] Reference 1: CN117441900B Summary of the Invention

[0010] The problem the invention aims to solve

[0011] Although existing technologies have been used to study the efficacy of casein phosphopeptides and their combinations, current research on the efficacy of casein phosphopeptides mainly focuses on promoting mineral absorption and regulating immunity. Therefore, there is still room for further exploration of new efficacy studies for casein phosphopeptides.

[0012] Therefore, the primary objective of this invention is to provide a novel use for casein phosphopeptides, specifically, to provide a use for casein phosphopeptides to help regulate tryptophan metabolism in the body.

[0013] Further research in this invention has demonstrated that casein phosphopeptides have a verifiable effect in regulating tryptophan metabolism in the body. This invention uses *C. elegans* as the research subject, analyzing and determining the content of tryptophan and its key metabolites within it, and demonstrating the regulation of tryptophan metabolism by casein phosphopeptides through the serotonin metabolic pathway and / or the kynurenine metabolic pathway within a certain concentration range of casein phosphopeptides.

[0014] Solution for solving the problem

[0015] The present invention has found that the above-mentioned technical problems can be solved by the following technical solutions:

[0016] [1]. Use of a casein phosphopeptide in the preparation of foods that help regulate tryptophan metabolism in the body.

[0017] [2]. According to the use described in [1], wherein the casein phosphopeptide includes one or more of casein phosphopeptides derived from human milk, casein phosphopeptides derived from cow milk, casein phosphopeptides derived from sheep milk, and casein phosphopeptides derived from casein.

[0018] [3]. According to the use described in [1] or [2], wherein the regulation of tryptophan metabolism in the body includes: increasing the tryptophan content in the body, and / or increasing the content of at least one metabolite in the tryptophan metabolic pathway.

[0019] [4]. According to the use described in [3], wherein the tryptophan metabolic pathway includes the serotonin metabolic pathway and / or the kynurenine metabolic pathway.

[0020] [5]. According to the use described in [3] or [4], wherein the metabolite is selected from at least one of the following: 5-methoxyindole-3-acetic acid, N-acetylserotonin, kynurenine, 2-pyridinecarboxylic acid, indole-3-carboxaldehyde.

[0021] [6]. According to any one of [1]-[5], wherein the food helps with mood regulation, sleep regulation and / or neuroprotection by regulating tryptophan metabolism in the body.

[0022] [7]. The use according to any one of [1]-[6], wherein the food is a powdered reconstituteable food, a baked food, a beverage, milk and dairy products or pasta products.

[0023] [8]. According to any one of [1]-[7], wherein, at room temperature, the food is in liquid form, solid-liquid mixture form, solid block form or solid powder form.

[0024] [9]. According to any one of [1]-[8], the food contains one or more of the following ingredients: plant-based ingredients, animal dairy ingredients, animal meat ingredients, functional additives and any food-acceptable excipients.

[0025]

[10] . According to any one of [1]-[9], wherein the content of casein phosphopeptide is 0.001% by mass or more based on the total mass of the food.

[0026] The effects of the invention

[0027] Based on the implementation of the above technical solution, the present invention can achieve the following technical effects:

[0028] 1) Experimental data from this invention show that casein phosphopeptides have the effect of regulating tryptophan metabolism in the body.

[0029] 2) Further research of this invention shows that casein phosphopeptides can increase the content of tryptophan and its key metabolites (including at least one of 5-methoxyindole-3-acetic acid, N-acetylserotonin, kynurenine, 2-pyridinecarboxylic acid, and indole-3-carboxaldehyde) in the body through the serotonin metabolic pathway and / or kynurenine metabolic pathway, thereby helping to exert the effects of mood regulation, sleep regulation and / or neuroprotection.

[0030] 3) This invention does not have any special requirements on the specific application of casein phosphopeptides or the form of food containing casein phosphopeptides, and is convenient to implement.

[0031] 4) This invention provides a novel application of casein phosphopeptides, offering new scientific basis and research ideas for their further development in functional foods and health products. In particular, there is currently no relevant research or application in the field regarding the novel efficacy of the casein phosphopeptides involved in this invention. No description or utilization of this novel efficacy has been found in currently published national standards, textbooks, papers, patents, or specific products. This provides space and necessity for the innovation of this invention. Through in-depth research on the novel efficacy of casein phosphopeptides, it is hoped that new products with unique functions can be developed to meet the market demand for new applications of functional ingredients. Attached Figure Description

[0032] Figure 1 The effect of different concentrations of casein phosphopeptide on tryptophan content in nematodes;

[0033] Figure 2 The effect of different concentrations of casein phosphopeptide on the content of 5-methoxyindole-3-acetic acid in nematodes;

[0034] Figure 3 The effect of different concentrations of casein phosphopeptide on the N-acetylserotonin content in nematodes;

[0035] Figure 4 The effect of different concentrations of casein phosphopeptide on kynurenine content in nematodes;

[0036] Figure 5 The effect of different concentrations of casein phosphopeptide on the content of 2-pyridinecarboxylic acid in nematodes;

[0037] Figure 6 The effect of different concentrations of casein phosphopeptide on the content of indole-3-carboxaldehyde in nematodes. Detailed Implementation

[0038] The following describes embodiments of the present invention, but the present invention is not limited thereto. The present invention is not limited to the various configurations described below, and various modifications can be made within the scope of the claims. Embodiments and examples obtained by appropriately combining the technical means disclosed in different embodiments and examples are also included in the technical scope of the present invention.

[0039] In this specification, the range of values ​​referred to as "value A to value B" refers to the range including the endpoint values ​​A and B.

[0040] Unless otherwise stated, in this instruction manual, "more" in "multiple", "multi-variety", "multiple", etc., means a value of 2 or more.

[0041] In this specification, the terms "comprising," "having," "including," or "containing" can mean included or open-ended, and do not exclude additional, uncited elements or method steps. At the same time, "comprising," "having," "including," or "containing" can also mean closed-ended, excluding additional, uncited elements or method steps.

[0042] In this specification, the numerical range indicated by "above" or "below" refers to the numerical range that includes the stated number.

[0043] In this specification, the word "may" has two meanings: to perform a certain process and not to perform a certain process.

[0044] In this specification, the terms "optional" or "optional" are used to indicate the use or omission of certain substances, components, procedures, application conditions, etc.

[0045] Unless otherwise specified, “room temperature” in this instruction manual usually refers to a temperature of 23±2℃.

[0046] All unit names used in this manual are international standard unit names, and unless otherwise stated, the "%" indicates weight or mass percentage.

[0047] In this specification, references to "some specific / preferred embodiments," "other specific / preferred embodiments," "implementation," etc., refer to specific elements (e.g., features, structures, properties, and / or characteristics) related to that embodiment, which are included in at least one of the embodiments described herein and may or may not be present in other embodiments. Furthermore, it should be understood that these elements may be combined in any suitable manner in various embodiments.

[0048] This invention primarily provides a novel use for casein phosphopeptides. Specifically, experimental research has revealed a beneficial effect of casein phosphopeptides related to the regulation of tryptophan metabolism in the body, and based on this research, it expands existing applications of casein phosphopeptides. This invention is mainly based on the following insights:

[0049] Using *Caenorhabditis elegans* as a model organism, this study systematically analyzed the changes in tryptophan and its metabolites in vivo. The experimental results showed that, within a specific concentration range, casein phosphopeptide can effectively regulate tryptophan metabolism. Its mechanism of action is as follows: casein phosphopeptide significantly increases tryptophan levels in the body through the serotonin metabolic pathway and / or the kynurenine metabolic pathway, and promotes the generation of key metabolites in the tryptophan metabolic pathway, including at least one of 5-methoxyindole-3-acetic acid, N-acetylserotonin, kynurenine, 2-pyridinecarboxylic acid, and indole-3-carboxaldehyde.

[0050] Based on the above-mentioned metabolic regulatory effects, this invention further clarifies that casein phosphopeptides exhibit clear application potential in mood regulation, sleep regulation, and neuroprotection, providing a solid scientific basis for their innovative application in functional foods and health products.

[0051] Casein phosphopeptide

[0052] Casein phosphopeptides (CPP) are bioactive polypeptides produced from bovine milk casein using biotechnology. They can be used in various nutritional and health foods and can effectively promote the absorption and utilization of divalent mineral nutrients such as calcium, iron, and zinc by the human body.

[0053] Casein phosphopeptide (CPP) is produced by hydrolyzing casein with trypsin, followed by purification and refinement. Its molecular structure consists of twenty to thirty amino acid residues, including four to seven clustered phosphoseryl groups. Currently, commercially available CPP products have a purity of 12%-90%.

[0054] In some embodiments, the casein phosphopeptides of the present invention include one or more of the following: casein phosphopeptides derived from human milk, casein phosphopeptides derived from bovine milk, casein phosphopeptides derived from goat milk, and casein-derived casein phosphopeptides. Casein phosphopeptides derived from bovine milk are preferred. Accordingly, casein phosphopeptides can be isolated from bovine milk or commercially available finished products can be purchased directly.

[0055] Furthermore, it should be emphasized that the above-mentioned form of addition of casein phosphopeptide must comply with the permitted scope of local laws and regulations. Within this permitted scope, it can be added as a single ingredient. When it is not permitted to be used as a single ingredient, other permitted components containing this ingredient can be used to impart the required casein phosphopeptide to the form of use. In addition, the content of casein phosphopeptide must also be configured within the permitted scope of local laws and regulations, and the content must be configured to be sufficient to cause the effects of the present invention.

[0056] In this invention, there are no particular limitations on the final form of casein phosphopeptides. At room temperature, they can be liquid (with the help of the desired solvent), semi-solid (water-containing slurry), solid (such as powder, granules or blocks (after pressing)), etc.

[0057] Foods containing casein phosphopeptides (including health foods)

[0058] The food containing casein phosphopeptides described in this invention can be in any form. In some specific embodiments, at room temperature, the food described in this invention is in liquid form, solid-liquid mixture form, solid block form, or solid powder form.

[0059] In addition to casein phosphopeptides, the food products described in this invention may include other optional ingredients, depending on the needs of the final product. Examples of such ingredients include:

[0060] Plant-based ingredients include fruits such as fig, pomegranate, kiwi, orange, tangerine, pineapple, strawberry, apple, banana, grape, pear, cherry, blueberry, blackberry, raspberry, and bilberry, or their extracts; fruits and vegetables such as onion, cucumber, tomato, cauliflower, carrot, spinach, kale, Brussels sprouts, garlic, basil, and oregano, or their extracts; grains such as rice (indica rice, japonica rice, glutinous rice), cereals (wheat, barley, oats, rye), corn, sorghum, millet, sorghum, yellow millet, buckwheat, soybean, broad bean, pea, mung bean, red bean, and kidney bean, or their extracts; nuts such as walnut, cashew, hazelnut, almond, apricot kernel, pine nut, pistachio, peanut, sunflower seed, chestnut, macadamia nut, and ginkgo, or their extracts; coffee or its extracts; and some medicinal and edible herbal medicines or their extracts.

[0061] Animal dairy products include fresh milk from cows, sheep, etc., as well as processed dairy products such as milk powder, whey protein, or cheese.

[0062] Animal meat ingredients, including meat ingredients from beef, mutton, fish or poultry.

[0063] Functional additives include vitamins (vitamin A, beta-carotene, vitamin D3, vitamin E, vitamin K1, vitamin B1, vitamin B2, vitamin B6, vitamin B1). 12 One or more of the following: Vitamin C, pantothenic acid, folic acid, niacin, choline, inositol, and biotin; starch; modified starch; amino acids (L-lysine-L-glutamic acid, L-glutamic acid, L-arginine, L-tryptophan, L-glutamine, taurine, L-valine, L-isoleucine, or L-leucine, etc.); dietary fiber (inulin, konjac flour, galactooligosaccharides, fructooligosaccharides, isomaltooligosaccharides, soybean polysaccharides, cyclodextrin, resistant dextrin, or soybean fiber, etc.); micronutrient supplements (including metal ions from organic acids). Salts, such as one or more of calcium citrate, L-calcium lactate, calcium hydrogen phosphate, potassium gluconate, sodium citrate, ferrous gluconate, zinc gluconate, sodium selenite, copper gluconate, manganese gluconate, and magnesium gluconate; fat supplements (such as saturated fatty acids, polyunsaturated fatty acids, monounsaturated fatty acids, OPO structured lipids, OPL structured lipids, LPL structured lipids, DHA, EPA, ARA, phospholipids, etc.); nucleotide supplements; human milk oligosaccharides (such as 2'-FL, 3-FL, DFL, LNFP I, LNFP II, LNT, LNnT, 3'-SL, 6'-SL, DSLNT, etc.), etc.

[0064] Any food additives that are acceptable, including but not limited to solvents, antioxidants, antibacterial agents, thickeners, diluents, solubilizers, stabilizers, emulsifiers, fillers, disintegrants, lubricants, coating materials, anti-caking agents, flavoring agents, sweeteners, food flavorings, and food colorings.

[0065] There is no particular limitation on the specific form of the food products of this invention. In some specific embodiments, the food products may be powdered instant food products (instant coffee, cereal powder, nut powder or lotus root powder, etc.), baked food products (bread, cake or biscuit baked food products, etc.), beverages (carbonated beverages, fruit and vegetable juice beverages, functional beverages, tea beverages or milk beverages, etc.), milk and dairy products (fresh milk, milk powder, whey powder, fermented milk, cheese or condensed milk, etc. derived from fresh cow (sheep) milk), pasta products (noodles, instant noodles, steamed buns, dumplings or wontons, etc.), etc.

[0066] In the food products described in this invention, the content of casein phosphopeptide complies with the requirements of relevant laws and regulations, and the content of casein phosphopeptide is sufficient to achieve the effective amount required to produce the technical effects involved in this invention. In some specific embodiments of this invention, the content of casein phosphopeptide, based on the total mass of the food product, is 0.001% by mass or more, preferably 0.005% by mass or more, and more preferably 0.04% by mass or more. There is no particular limit to the upper limit of its content, and reference can be made to the regulations of different regions or conventional usage.

[0067] Uses for regulating tryptophan metabolism in the body

[0068] This invention reveals for the first time a novel application of casein phosphopeptides in regulating tryptophan metabolism in the body / human body. Based on this discovery, foods containing casein phosphopeptides are also suitable for this application, and their consumption can also help to positively regulate tryptophan metabolism in the body.

[0069] In some specific implementations, the regulation of tryptophan metabolism includes: increasing the tryptophan content in the body, and / or increasing the content of at least one metabolite in the tryptophan metabolic pathway.

[0070] In some specific implementations, the tryptophan metabolic pathway includes the serotonin metabolic pathway and / or the kynurenine metabolic pathway.

[0071] In some specific embodiments, the metabolite is selected from at least one of the following: 5-methoxyindole-3-acetic acid, N-acetylserotonin, kynurenine, 2-pyridinecarboxylic acid, and indole-3-carboxaldehyde.

[0072] In some specific implementations, the food helps regulate mood, sleep, and / or neuroprotection by modulating tryptophan metabolism in the body.

[0073] In addition, the food of the present invention is suitable for all people in principle, especially for people with specific needs for mood management, sleep quality or neurological health. The components of the food can also be adjusted accordingly for people with different characteristics.

[0074] Example

[0075] The embodiments of the present invention will be described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered as limiting the scope of the invention. Unless otherwise specified in the examples, conventional conditions or conditions recommended by the manufacturer are followed. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0076] Example 1: Effects of CPP on tryptophan and its metabolites in nematodes

[0077] 1. Method

[0078] (1) Solution preparation

[0079] 1) Nematode lysis solution (Clorox solution): 5% NaClO: 1M NaOH = 2:1 (v / v), NaClO is treated in the dark.

[0080] 2) 1M potassium phosphate buffer (pH 6.0): 108.3g KH2PO4, 35.6g K2HPO4 (46.63g K2HPO4·3H2O), 1L water, autoclaved for later use.

[0081] 3) S buffer: 129 mL 0.05 M K2HPO4 (1.12 g), 871 mL 0.05 M KH2PO4 (5.92 g), 5.85 g NaCl.

[0082] 4) Nematode cryopreservation solution: S buffer + 30% glycerol (volume ratio 1:1).

[0083] 5) 1M MgSO4: 120.3676g MgSO4 (246.47g MgSO4·7H2O), 1L water, filtered through a 0.22μm filter and set aside.

[0084] 6) 1M CaCl2: 110.984g CaCl2, 1L water, filtered through a 0.22μm filter and set aside.

[0085] 7) 5 mg / mL cholesterol solution: Dissolve 0.5 g cholesterol in 100 mL ethanol, filter through a 0.22 μm filter, and store at 4 °C for later use.

[0086] 8) Solid NGM medium: Dissolve 3g NaCl, 2.5g peptone, and 17g agar in 975mL of water, then sterilize at 121℃ for 15min. After sterilization, cool to 60℃, add 1mL 1M CaCl2, 1mL 1M MgSO4, 1mL 5mg / mL cholesterol solution, and 25mL 1M potassium phosphate buffer, pour into plates while hot, and store at 4℃ for later use.

[0087] 9) M9 buffer: 6g Na2HPO4, 3g KH2PO4, 5g NaCl, 1L water, sterilized and then add 1mL 1M MgSO4.

[0088] (2) Cryopreservation of Caenorhabditis elegans

[0089] Select plates that have just depleted E. coli OP50 bacterial colonies and contain a large number of L1-L2 stage nematodes. Collect the nematodes into 1.5 mL centrifuge tubes using M9 buffer, centrifuge to remove the supernatant, centrifuge at 3000 rpm for 3 min, discard the supernatant, add 1 mL of cryopreservation solution, vortex to mix, transfer to 1.8 mL cryovials, label with bacterial strain and date, store at -4℃ for 1 day, at -20℃ for 1 day, and at -80℃.

[0090] (3) Resuscitation of Caenorhabditis elegans

[0091] After thawing the frozen nematode samples at room temperature, centrifuge at 2500 rpm for 2 min, discard the supernatant, and use a pipette to aspirate the precipitate at the bottom of the test tube and spread it onto NGM solid culture medium containing OP50.

[0092] (4) Preparation of OP50 seed solution

[0093] Inoculate OP50 from the slant into 100 mL of LB liquid medium, incubate overnight at 37°C and 220 rpm, and store at 4°C for later use.

[0094] (5) Production of NGM containing OP50

[0095] Take 100 μL (50 μL for a 35 mm plate) of OP50 bacterial suspension and spread it evenly on a 60 mm NGM plate. Dry at 37°C for 8 hours. Store in a tightly closed container.

[0096] (6) Synchronization of Caenorhabditis elegans

[0097] Add 1.2 mL of M9 buffer to NGM medium containing nematodes. Gently agitate the culture dish to allow the nematodes to detach from the medium and enter the buffer. Use a pipette to transfer 1 mL of M9 buffer containing nematodes into a 1.5 mL centrifuge tube. Centrifuge (3000 rpm, 1 min), remove the supernatant, add 1 mL of lysis buffer, and vortex to mix. Process for 10 min, centrifuge (3000 rpm, 1 min), and remove the supernatant. Then wash three times with M9 buffer to obtain nematode eggs.

[0098] (7) Sample preparation

[0099] CPP was dissolved in sterile water to prepare a 50 mg / mL stock solution, which was stored at 4°C protected from light. Before use, the stock solution was mixed with OP50 and diluted to working solutions with concentrations of 0.2, 1, and 2 mg / mL. The OP50 bacterial suspension containing CPP was then spread onto NGM plates and dried at 37°C for 8 hours. L1 stage larvae were transferred to NGM plates containing CPP (0.2, 1, and 2 mg / mL) (experimental group) and NGM plates containing only OP50 (control group), cultured for 48 hours, and nematode samples were collected for relevant testing.

[0100] (8) Metabolite detection

[0101] 50 μL of nematodes treated for 48 h were collected in a grinding tube, and the tissue was ground and broken up. Then, a 1:4 (v / v) volume of 50% acetonitrile solution with a concentration of 1% formic acid was added, and the mixture was sonicated for 10 min and vortexed for 2 min. Then, it was sonicated in an ice bath for 20 min, frozen at -20℃ for 60 min, and finally centrifuged at -4℃ (12000 rpm, 30 min). The supernatant was collected, freeze-dried, and reconstituted with 200 μL of 50% acetonitrile solution with a concentration of 1% formic acid. Finally, the mixture was filtered through a 0.22 μm organic filter membrane, and the neurotransmitter content was detected by liquid chromatography-mass spectrometry (LC-MS). LC-MS chromatographic analysis conditions: Discovery® HS F5-3 column (15cm × 2.1mm, 3μm); mobile phase A (0.1% formic acid aqueous solution), B (acetonitrile solution containing 0.1% formic acid); gradient elution (0-2min, 2% B; 2-10min, 2%-28% B; 10-13min, 28%-100% B; 13-14min, 100% B; 14-16min, 2% B); flow rate 0.40mL / min, column temperature 35℃, injection volume 2μL.

[0102] 2. Results

[0103] 2.1 Effect of CPP on tryptophan content in nematodes

[0104] The effect of CPP on tryptophan content in nematodes, such as Figure 1 As shown, under CPP concentrations of 1–2 mg / mL, the tryptophan content in nematodes was positively correlated with the CPP treatment concentration, exhibiting a significant dose-response effect.

[0105] 2.2 Effect of CPP on the content of 5-methoxyindole-3-acetic acid in nematodes

[0106] The effect of CPP on the 5-methoxyindole-3-acetic acid content in nematodes, such as Figure 2 As shown, under CPP concentrations of 1–2 mg / mL, the content of 5-methoxyindole-3-acetic acid in nematodes was positively correlated with the CPP treatment concentration, exhibiting a good dose-response effect.

[0107] 2.3 Effect of CPP on N-acetylserotonin (NAS) levels in nematodes

[0108] The effect of CPP on N-acetylserotonin levels in nematodes, such as Figure 3 As shown, compared with the control group, all CPP treatments significantly increased the content of N-acetylserotonin in nematodes (p < 0.05), and the effect showed a certain dose-dependent effect with increasing CPP concentration, indicating that different CPP concentrations can significantly increase the production of NAS in nematodes.

[0109] 2.4 Effect of CPP on kynurenine content in nematodes

[0110] The effect of CPP on kynurenine content in nematodes, such as Figure 4 As shown, higher concentrations of CPP can increase the level of kynurenine in nematodes.

[0111] 2.5 Effect of CPP on 2-pyridinecarboxylic acid content in nematodes

[0112] The effect of CPP on the 2-pyridinecarboxylic acid content in nematodes, such as Figure 5 As shown, treatment with different concentrations of CPP significantly increased the content of 2-pyridinecarboxylic acid in nematodes (p < 0.05).

[0113] 2.6 Effect of CPP on the content of indole-3-carboxaldehyde (I3A) in nematodes

[0114] The effect of CPP on I3A content in nematodes, such as Figure 6 As shown, treatment with medium to high concentrations (1-2 mg / mL) of CPP can significantly increase the content of I3A in nematodes.

[0115] 3. Discussion and Conclusion

[0116] Tryptophan, an essential amino acid, is the sole precursor in the biosynthesis of serotonin (5-HT) and melatonin. N-acetylserotonin (NAS) is an intermediate metabolite linking serotonin and melatonin synthesis and is also a direct agonist of the tropomyosin-associated kinase B (TrkB) receptor, playing a crucial role in mood regulation. Through this mechanism, NAS mimics the function of brain-derived neurotrophic factor (BDNF), exhibiting significant antidepressant and neuroprotective activity. Furthermore, NAS links the circadian rhythm of nocturnal melatonin synthesis to hippocampal plasticity and neurogenesis during sleep, participating not only in mood regulation but also possessing potent antioxidant and anti-apoptotic capabilities. Our results show that CPP significantly increases tryptophan and NAS levels in nematodes, suggesting that CPP may have the potential to regulate mood, improve sleep, and protect the nervous system.

[0117] Aside from generating serotonin, the vast majority of tryptophan (approximately 95%) is metabolized via the kynurenine pathway (KP). Kynurenine has neuroprotective effects, the mechanisms of which include: directly scavenging reactive oxygen species such as superoxide anions and hydroxyl radicals, reducing free radical damage to neurons; regulating inflammatory responses, reducing the release of pro-inflammatory cytokines, thereby alleviating neuronal damage caused by inflammation; and participating in the regulation of neuronal energy metabolism, maintaining its structure and function.

[0118] 2-Pyridinecarboxylic acid, as a downstream metabolite of the kynurenine pathway, can inhibit the Fenton reaction by chelating metal ions such as iron and copper, reducing the production of reactive oxygen species (ROS). It can also activate the expression of downstream antioxidant enzyme genes, increasing the activity of enzymes such as superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), thereby constructing a multi-layered antioxidant defense system to protect neurons from oxidative damage. On the other hand, indole-3-carboxaldehyde (I3A) has been shown to have certain neuroprotective effects in recent years. Its mechanism may involve multiple aspects such as antioxidation, anti-inflammation, and regulation of neurotransmitters, for example, alleviating oxidative stress damage by scavenging intraneural ROS.

[0119] In summary, the results of this study indicate that CPP intervention can significantly increase the levels of tryptophan and its key metabolites in nematodes, suggesting that CPP has the ability to regulate tryptophan metabolism and may therefore play a role in mood and sleep regulation. At the same time, the regulation of the kynurenine pathway by CPP also suggests that it has certain neuroprotective potential.

[0120] It should be noted that although the technical solution of the present invention has been described with specific examples, those skilled in the art will understand that the present invention should not be limited thereto.

[0121] The various embodiments of the present invention have been described above. These descriptions are exemplary and not exhaustive, nor are they limited to the disclosed embodiments. Many modifications and variations will be apparent to those skilled in the art without departing from the scope and spirit of the described embodiments. The terminology used herein is chosen to best explain the principles, practical application, or technical improvements to the embodiments in the market, or to enable others skilled in the art to understand the embodiments disclosed herein.

Claims

1. Use of a casein phosphopeptide in the preparation of foods that help regulate tryptophan metabolism in the body.

2. The use according to claim 1, characterized in that, The regulation of tryptophan metabolism includes: increasing the tryptophan content in the body, and / or increasing the content of at least one metabolite in the tryptophan metabolic pathway.

3. The use according to claim 2, characterized in that, The tryptophan metabolic pathway includes the serotonin metabolic pathway and / or the kynurenine metabolic pathway.

4. The use according to claim 2 or 3, characterized in that, The metabolite is selected from at least one of the following: 5-methoxyindole-3-acetic acid, N-acetylserotonin, kynurenine, 2-pyridinecarboxylic acid, and indole-3-carboxaldehyde.

5. The use according to any one of claims 1-4, characterized in that, The food products mentioned are powdered reconstituteable foods, baked goods, beverages, milk and dairy products, or pasta products.

6. The use according to any one of claims 1-5, characterized in that, At room temperature, the food is in liquid form, solid-liquid mixture form, solid block form, or solid powder form.

7. The use according to any one of claims 1-6, characterized in that, The food contains one or more of the following ingredients: plant-based ingredients, animal dairy ingredients, animal meat ingredients, functional additives, and any food-acceptable excipients.

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