Composition and application thereof in preparation of medicines and health foods for improving sleep

Abstract

The invention discloses a composition and application thereof in preparation of drugs and health food for improving sleep, the composition comprises the components of ganoderan, casein hydrolysate peptide and spina date seed powder, any two components can be compounded according to a certain proportion, and the three components can also be compounded according to a certain proportion. The three compounded compositions comprise the following components in parts by weight: 0.5-9 parts of ganoderma lucidum polysaccharide; 0.3 to 9 parts of casein hydrolysate peptide; and 0.5-9 parts of spina date seed powder. In the composition compounded by the ganoderma lucidum polysaccharide and the spina date seed powder, the weight ratio of the ganoderma lucidum polysaccharide to the spina date seed powder is (1: 9)-(9: 1). Through verification, the composition disclosed by the invention has a remarkable synergistic effect in the aspects of sleep round times, total sleep duration and average sleep duration, and is simple in components, safe in source, easy to prepare and easy to industrially produce.

Description

Technical Field

[0001] This invention relates to the fields of biomedicine and health food, specifically to a composition and its application in the preparation of sleep-improving drugs and health foods. Background Technology

[0002] With the fast pace of modern life, sleep problems are becoming increasingly prominent, mainly manifested as late bedtime, difficulty falling asleep, insufficient sleep duration, and poor sleep quality. Sleep problems not only affect daily life and study, but may also induce a variety of health problems.

[0003] In recent years, numerous patents and documents have reported on natural small molecule compounds, traditional Chinese medicine compositions, health food compositions, and natural food ingredients that improve sleep. For example, patent document CN 105902536A discloses the application of loganin in the preparation of drugs or health products for improving sleep. Animal experiments have confirmed that loganin can prolong the total sleep time of mice and increase the amount of sleep in a dose-dependent manner. Patent document CN102309565A discloses a traditional Chinese medicine composition for improving sleep and its preparation method. This traditional Chinese medicine composition includes the following raw materials in parts by weight: 150-250 parts of jujube seed extract, 100-150 parts of cypress seed extract, 100-150 parts of processed Polygonum multiflorum extract, 100-120 parts of Epimedium extract, 100-150 parts of jujube extract, 100-150 parts of Acanthopanax senticosus extract, and 5 parts of vitamin B11.

[0004] However, single components have limited improvement effects, chemical drugs have many side effects, and combinations have problems such as complex components, complicated preparation, or unknown compatibility interactions. Summary of the Invention

[0005] This application provides a composition and its application in the preparation of sleep-improving drugs and health foods. The composition has a significant synergistic effect in terms of the number of sleep rounds, total sleep duration and average sleep duration. Moreover, the composition is simple in composition, safe in source, easy to prepare, and easy to industrially produce.

[0006] This application constructs an insomnia model by inducing abnormal sleep behavior in zebrafish juveniles using pentylenetetrazol. A behavioral analyzer is used to evaluate the effects of two or more combinations of hydrolyzed casein peptides, Ganoderma lucidum polysaccharides, jujube seed powder, longan, asparagus, and lily on sleep phenotypes (sleep improvement rate, number of sleep rounds, sleep duration, and average sleep duration), and compares them with the sleep-aiding effects of the recognized sleep-improving hormone melatonin.

[0007] The synergistic effect of the composition on sleep improvement was quantitatively analyzed using the combined index method. Finally, using diazepam as a positive modeling drug, animal experiments based on mice were conducted to verify the synergistic effect of the formulation on sleep behavior and its influence on sleep neurotransmitter and hormone levels, thus validating the sleep improvement effect. Finally, based on human trial data, a composition with significant synergistic sleep improvement was developed.

[0008] The composition comprises Ganoderma lucidum polysaccharides, hydrolyzed casein peptides, and jujube seed powder. Any two of these components can be combined in a specific ratio, or all three components can be combined in a specific proportion. Compositions with specific ratios consistently demonstrate better sleep improvement rates than melatonin, and their sleep frequency, sleep duration, and average sleep duration are close to those of melatonin.

[0009] The polysaccharides and triterpenoids in Ganoderma lucidum improve sleep through multiple mechanisms. Specifically, Ganoderma lucidum polysaccharides can regulate the nervous system, promote melatonin secretion from the pineal gland, shorten sleep latency, and increase sleep duration. Hydrolyzed casein peptides can increase the content of the melatonin precursor serotonin in the brain, significantly shortening sleep latency. Hydrolyzed casein peptides can also enhance the effect of the inhibitory neurotransmitter γ-aminobutyric acid (GABA) by binding to GABA receptors, while simultaneously reducing the level of the excitatory neurotransmitter glutamate, inhibiting nervous system excitation, and promoting deep sleep. Jujube seed powder contains jujube seed extract, a core component of the traditional Chinese medicine jujube seed. Its sleep-improving mechanism mainly involves multiple pathways, including regulating neurotransmitters, inhibiting excessive activation of the hypothalamus-pituitary-adrenal axis, anti-inflammation, neuronal protection, and promoting neuronal health. The sedative-hypnotic effects of jujube seed extract are not only suitable for insomnia patients but also have an adjunctive therapeutic effect on neurasthenia, anxiety, and other diseases.

[0010] This application verifies the sleep-improving effect of the composition of the present invention in accordance with the standard method of "Methods for Testing and Evaluating the Functions of Health Products". The results show that the compound powder of Ganoderma lucidum polysaccharide, jujube seed powder and hydrolyzed casein peptide, by increasing the level of 5-hydroxytryptamine and the γ-aminobutyric acid / glutamate ratio, and decreasing the level of the excitatory neurotransmitter glutamate, significantly shortens the sleep latency, increases the sleep duration, and improves the sleep of mice, and has been proven to have a significant sleep-improving effect.

[0011] This application also verified the sleep-improving effect of a pilot-scale beverage with the composition of this application as the core formula by recruiting volunteers. The results showed that after taking the compound beverage of Ganoderma lucidum polysaccharide, jujube seed powder and hydrolyzed casein peptide for 7 days, 95% of the participants experienced a shorter time to fall asleep, 84% experienced a longer total sleep time, 79% experienced improved sleep quality, and 89% experienced improved mental state after waking up. By plotting the 7-day data of the test group into a curve, it was found that with the extension of the treatment time, the participants' time to fall asleep, total sleep time, sleep quality and mental state after waking up all showed continuous improvement.

[0012] Based on the above research results, this application first provides a composition for improving sleep. The composition is a compound product of three components: Ganoderma lucidum polysaccharide, hydrolyzed casein peptide, and jujube seed powder. By weight, its components are: 0.5-9 parts of Ganoderma lucidum polysaccharide; 0.3-9 parts of hydrolyzed casein peptide; and 0.5-9 parts of jujube seed powder.

[0013] The components in the composition can be obtained commercially. The components obtained commercially are all in powder form, usually water extracts of the corresponding raw materials. The preparation method can be conventional mixing, such as uniformly mixing the powdered raw materials. For a composition of three components, two components can be mixed in a specific ratio first, and then mixed with the third component in a specific ratio.

[0014] Each component in the composition can also be prepared by conventional processing methods in the art: the raw materials corresponding to the components, such as Ganoderma lucidum, casein and jujube seed, are obtained by solvent extraction, concentration / drying and other methods, and then compounded; or the raw materials can be weighed in a certain proportion and physically mixed, and then obtained by solvent extraction, concentration / drying and other methods.

[0015] The preparation method of Ganoderma lucidum polysaccharide is as follows: qualified Ganoderma lucidum is pulverized into ultrafine powder, and water or low proportion of ethanol is used as solvent. Then, according to a suitable material-liquid ratio (1:10-1:30), it is extracted in a multi-functional extraction tank (100-150 ℃, 2 hours), plate and frame filtered, membrane concentrated, and freeze-dried to obtain powder, which is used as a compound raw material in the following examples.

[0016] The preparation method of jujube seed powder is as follows: qualified jujube seeds are crushed into ultrafine powder, and water or low proportion ethanol is used as solvent. Then, the powder is extracted in a multi-functional extraction tank at a suitable material-liquid ratio (1:10-1:20) (50℃, 1 hour), filtered through gauze, concentrated through membrane, and freeze-dried to obtain powder, which is used as a compound raw material in the following examples.

[0017] The preparation method of hydrolyzed casein peptides is as follows: using water as a solvent, casein is added at a suitable material-to-liquid ratio (usually 1:10-1:100) with 1-10% of a complex enzyme (neutral protease, bromelain, and flavor enzyme, etc., to be formulated as needed) in an enzymatic hydrolysis tank for 4-6 hours (pH 5.0-8.5, 45-65 ℃). After enzyme inactivation at high temperature (90 ℃, 15 minutes), the peptides are then obtained by centrifugation, concentration, and spray drying.

[0018] Regardless of whether it is obtained through commercial purchases or self-processing, its components should preferably meet the following conditions: The content of Ganoderma lucidum polysaccharides in Ganoderma lucidum polysaccharides is greater than 5%; the proportion of hydrolyzed casein peptides with a relative molecular mass of less than 1000 Da is not less than 80%, and the proportion of oligopeptides (molecular weight less than 3000 Da) is not less than 90%; the content of total jujuboside in jujube seed powder is not less than 1%.

[0019] A preferred composition comprising three components, wherein the hydrolyzed casein peptide and jujube seed powder are compounded at a weight ratio of 1:2, and then compounded with Ganoderma lucidum polysaccharide at a weight ratio of 9:1 to 1:9 based on their total weight. More preferably, the hydrolyzed casein peptide and jujube seed powder are compounded at a weight ratio of 1:2, and then compounded with Ganoderma lucidum polysaccharide at a weight ratio of 9:1 or 1:9 based on their total weight.

[0020] Another preferred composition is a three-component compound, wherein the Ganoderma lucidum polysaccharide and jujube seed powder are compounded at a weight ratio of 1:11, and then compounded with hydrolyzed casein peptides at a weight ratio of 9:1 to 1:9 based on their total weight. More preferably, the Ganoderma lucidum polysaccharide and jujube seed powder are compounded at a weight ratio of 1:11, and then compounded with hydrolyzed casein peptides at a weight ratio of 9:1 or 1:9 based on their total weight.

[0021] Another preferred composition is a three-component compound, wherein the Ganoderma lucidum polysaccharide and hydrolyzed casein peptide are compounded at a weight ratio of 2:1, and then compounded with the hydrolyzed casein peptide at a weight ratio of 9:1 to 1:9 of the total weight of the two components and jujube seed powder. More preferably, the Ganoderma lucidum polysaccharide and hydrolyzed casein peptide are compounded at a weight ratio of 2:1, and then compounded with the hydrolyzed casein peptide at a weight ratio of 9:1 or 1:9 of the total weight of the two components and jujube seed powder.

[0022] The composition of this application may also be a combination of any two components: An optional pairwise compounding scheme is provided, wherein the Ganoderma lucidum polysaccharide and jujube seed powder are compounded in a weight ratio of 1:9 to 9:1. Further, the Ganoderma lucidum polysaccharide and jujube seed powder are compounded in a weight ratio of 1:9, 1:1, or 9:1. When the weight ratio of Ganoderma lucidum polysaccharide and jujube seed powder is 1:9, the improvement effect is close to that of melatonin; when the weight ratio of Ganoderma lucidum polysaccharide and jujube seed powder is 9:1, the improvement effect is superior to that of melatonin.

[0023] Another optional pairwise compounding scheme involves combining Ganoderma lucidum polysaccharides and hydrolyzed casein peptides in a weight ratio of 1:4 to 4:1. Further, the Ganoderma lucidum polysaccharides and hydrolyzed casein peptides are compounded in a weight ratio of 1:4, 2:1, or 4:1.

[0024] Another optional pairwise compounding scheme is that the jujube seed powder and hydrolyzed casein peptides are compounded in a weight ratio of 1:4 to 4:1. Further, the jujube seed powder and hydrolyzed casein peptides are compounded in a weight ratio of 1:4, 2:1, or 4:1.

[0025] Based on the efficacy of the composition verified in this application, this application also provides an application of the composition in the preparation of sleep-improving products.

[0026] Optionally, the improvement of sleep includes, but is not limited to, increasing the sleep improvement rate, increasing the number of sleep rounds, sleep duration, and average sleep duration.

[0027] Optionally, the sleep-improving product may be a pharmaceutical or health food product.

[0028] When used in the preparation of health food products, the composition can be prepared with food-acceptable excipients such as solvents, binders, fillers, sugars, and sweeteners according to conventional food preparation methods. The type of health food product can be any of the following: tablets, capsules, oral liquids, solid beverages, compressed candies, pills, granules, decoctions, pastes, syrups, drop pills, or tea bags. Optionally, the health food product is a solid beverage, compressed candy, or oral liquid.

[0029] More preferably, when the composition of the composition by weight is: 9 parts of Ganoderma lucidum polysaccharide and hydrolyzed casein peptide compound powder (2:1), and 1 part of jujube seed powder; the oral liquid contains: Ganoderma lucidum polysaccharide 7.6 mg / mL, jujube seed powder 1.2 mg / mL, and hydrolyzed casein peptide 3.6 mg / mL.

[0030] When used in pharmaceutical preparation, the composition can be prepared with pharmaceutically acceptable excipients according to conventional pharmaceutical preparation methods. Pharmaceutically acceptable excipients include buffer solutions, emulsifiers, suspending agents, disintegrants, disintegrants, dispersants, binders, excipients, stabilizers, chelating agents, diluents, gelling agents, preservatives, wetting agents, lubricants, absorption delay agents, liposomes, etc., which can be selected according to different needs. The dosage forms of the pharmaceutical product include granules, tablets, capsules, powders, oral liquids, pellets, microcapsules, or injections.

[0031] In the aforementioned health foods and pharmaceuticals, the amount and proportion of the composition and related excipients of the present invention can be adjusted according to actual needs.

[0032] Compared with existing products, this application has the following advantages: The composition of this application has been proven to have significant sleep-improving effects. Ganoderma lucidum polysaccharide, hydrolyzed casein peptide and jujube seed powder have a synergistic effect, showing significant improvement in the number of sleep rounds, total sleep duration and average sleep duration, which is close to or better than the positive control. Moreover, the composition is simple, safe in origin, easy to prepare and easy to industrially produce. Attached Figure Description

[0033] Figure 1The graph shows the results of the volunteer group in Experiment Example 7, including shortened sleep onset time (A), prolonged total sleep time (B), improved sleep quality (C), and improved mental state after waking up (D).

[0034] Figure 2 The graph shows the improvement in sleep onset time (A), total sleep time (B), sleep quality (C), and mental state after waking up (D) for the volunteer group in Example 7 after 7 consecutive days of use. Detailed Implementation

[0035] The technical solution of this application will be clearly and completely described below with reference to specific embodiments. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments in this application, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of this application.

[0036] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

[0037] The Ganoderma lucidum polysaccharides (extract, with a content of more than 5%), hydrolyzed casein peptides (with a relative molecular mass of less than 1000 Da and a proportion of less than 80% of the protein hydrolysates, and a proportion of less than 90% of the oligopeptides (with a molecular weight of less than 3000 Da), and jujube seed powder (with a total jujube seed glycoside content of less than 1%) were all purchased from the market.

[0038] Example 1 The composition of this embodiment, by weight, consists of the following components: 1 part Ganoderma lucidum polysaccharide and 1 part Ziziphus jujuba seed powder.

[0039] Example 2 The composition of this embodiment, by weight, consists of the following components: 1 part Ganoderma lucidum polysaccharide and 9 parts Ziziphus jujuba seed powder.

[0040] Example 3 The composition of this embodiment, by weight, consists of the following components: 9 parts Ganoderma lucidum polysaccharide and 1 part Ziziphus jujuba seed powder.

[0041] Example 4 The composition of this embodiment, by weight, consists of the following components: 2 parts Ganoderma lucidum polysaccharide and 1 part hydrolyzed casein peptide.

[0042] Example 5 The composition of this embodiment, by weight, consists of the following components: 2 parts Ganoderma lucidum polysaccharide and 8 parts hydrolyzed casein peptide.

[0043] Example 6 The composition of this embodiment, by weight, consists of the following components: 8 parts Ganoderma lucidum polysaccharide and 2 parts hydrolyzed casein peptide.

[0044] Example 7 The composition of this embodiment, by weight, consists of the following components: 2 parts jujube seed powder and 1 part hydrolyzed casein peptide.

[0045] Example 8 The composition of this embodiment, by weight, consists of the following components: 2 parts jujube seed powder and 8 parts hydrolyzed casein peptide.

[0046] Example 9 The composition of this embodiment, by weight, consists of the following components: 8 parts jujube seed powder and 2 parts hydrolyzed casein peptide.

[0047] Example 10 The composition of this embodiment, by weight, consists of the following components: 9 parts hydrolyzed casein peptide and jujube seed powder compound powder (1:2), and 1 part Ganoderma lucidum polysaccharide.

[0048] Example 11 The composition of this embodiment, by weight, consists of the following components: 1 part hydrolyzed casein peptide and jujube seed powder compound powder (1:2), and 9 parts Ganoderma lucidum polysaccharide.

[0049] Example 12 The composition of this embodiment, by weight, consists of the following components: 9 parts of Ganoderma lucidum polysaccharide and jujube seed powder compound powder (1:1), and 1 part of hydrolyzed casein peptide.

[0050] Example 13 The composition of this embodiment, by weight, consists of the following components: 1 part of Ganoderma lucidum polysaccharide and jujube seed powder compound powder (1:1), and 9 parts of hydrolyzed casein peptide.

[0051] Example 14 The composition of this embodiment, by weight, consists of the following components: 9 parts of Ganoderma lucidum polysaccharide and hydrolyzed casein peptide compound powder (2:1), and 1 part of jujube seed powder.

[0052] Example 15 The composition of this embodiment, by weight, consists of the following components: 1 part of Ganoderma lucidum polysaccharide and hydrolyzed casein peptide compound powder (2:1), and 9 parts of jujube seed powder.

[0053] Experiment 1 (Raw Material Screening) Wild-type AB strain zebrafish with a 6 dpf growth rate were selected, and an insomnia model was induced using pentylenetetrazol. Melatonin was used as a positive control, and the melatonin concentration was set at 100 μM. Before the start of the behavioral experiment, pentylenetetrazol solution was added to the culture water of each group, and melatonin stock solution and treatment group stock solution were added dropwise in a timely manner to ensure that the dosage met the experimental setting. The compound raw materials are shown in Table 1.

[0054] Among them, jujube seed powder (EC50 is 210 μg·mL) -1 Hydrolyzed casein peptides (EC50: 114.5 μg·mL) -1 ) and Ganoderma lucidum polysaccharide (extract, EC50 164.6 μg·mL) -1 Dosage should be set according to their respective EC50 values; the dosage of asparagus powder is set at 750 μg·mL. -1 The dosage of longan powder was set at 550 μg / mL. -1 The dosage of lily powder was set at 600 μg / mL. -1 .

[0055] Zebrafish trajectories were recorded for 1 hour using a behavioral analyzer. Movements below 0.5 cm / s were considered slow, 0.5–2 cm / s were medium, and movements exceeding 2 cm / s were high. The total number of sleep episodes within 1 hour was recorded as the number of sleep cycles. A single successful sleep episode was defined as a slow movement lasting 6 seconds or more. Sleep duration was the total sleep time within 1 hour. Average sleep duration was the average sleep duration within 1 hour.

[0056] The sleep improvement rates are as follows: Sleep improvement rate = 100 × (total high-speed movement distance of the model group - total high-speed movement distance of the treatment group) / total high-speed movement distance of the model group.

[0057] The specific results are as follows: Table 1. Screening of raw materials for sleep-improving compound formulations based on zebrafish insomnia model.

[0058] Table 1 shows that the model group had 15.6 sleep cycles, a sleep duration of 274.8 s, and an average sleep duration of 17.6 s, all significantly lower than the normal group. These results are consistent with numerous literature reports on pentylenetetrazole-induced insomnia phenotypes in zebrafish. Meanwhile, the groups treated with the modeling drug and melatonin showed a sleep improvement rate of 76.2%, with 25.6 sleep cycles, a sleep duration of 728.5 s, and an average sleep duration of 28.4 s. This indicates that melatonin can reverse the pentylenetetrazole-induced insomnia phenotype. Therefore, these results demonstrate the successful establishment of a pentylenetetrazole-induced zebrafish insomnia model.

[0059] Based on the established zebrafish insomnia model, the effects of six food ingredients—Ganoderma lucidum polysaccharide, jujube seed powder, hydrolyzed casein peptide, asparagus powder, longan powder, and lily powder—on improving the insomnia phenotype were evaluated.

[0060] In terms of sleep improvement rate, the order of improvement effect is as follows: hydrolyzed casein peptide > Ganoderma lucidum polysaccharide > jujube seed powder > longan powder > asparagus powder > lily powder.

[0061] In terms of the number of sleep cycles, the order of improvement effect is as follows: hydrolyzed casein peptide > jujube seed powder > longan powder > Ganoderma lucidum polysaccharide > lily powder > asparagus powder.

[0062] In terms of sleep duration, the order of improvement effect is as follows: jujube seed powder > Ganoderma lucidum polysaccharide > hydrolyzed casein peptide > longan powder > lily powder > asparagus powder.

[0063] As shown in Table 1, both hydrolyzed casein peptides and Ganoderma lucidum polysaccharides significantly improved sleep quality, suggesting a potential synergistic effect when combined. Both hydrolyzed casein peptides and jujube seed powder significantly increased the number of sleep cycles, indicating a potential synergistic effect when combined. Both jujube seed powder and Ganoderma lucidum polysaccharides significantly prolonged sleep duration, suggesting a potential synergistic effect when combined.

[0064] Considering the effects and contributions of six food ingredients on sleep phenotype improvement, Ganoderma lucidum polysaccharides, jujube seed powder, and hydrolyzed casein peptides were selected as the core ingredients of the sleep-improving compound formula. Finally, based on the experimental confirmation of significant synergy between two ingredients, a third ingredient was added to further demonstrate the synergistic effect of the three ingredients. This invention aims to develop a synergistic sleep-improving compound formula; subsequent embodiments will be based on these projections.

[0065] Experiment Example 2 The experimental method is the same as in Experiment 1. The compound raw materials and results are shown in Table 2. Table 2. Effects of Ganoderma lucidum polysaccharide and jujube seed powder complex on sleep in zebrafish.

[0066] Note: The dosages used in Examples 1, 2, and 3 are all 50% effective doses; " / " indicates no numerical value; statistical differences compared to the model group are marked with *. p <0.05,** p <0.01, *** p <0.001.

[0067] Table 2 shows that the sleep duration of zebrafish treated with Ganoderma lucidum polysaccharide and jujube seed powder alone increased by 30.5% and 41.7% respectively compared with the model group. Based on the EC50 values ​​(half-maximal effective concentration) of Ganoderma lucidum polysaccharide and jujube seed powder during the raw material screening stage, the compound ratio of Ganoderma lucidum polysaccharide and jujube seed powder was set to 1:1, 1:9 and 9:1, with corresponding CI values ​​of 0.79, 0.69 and 0.62, respectively, all less than 1. This indicates that when the compound ratio is 1:9~9:1, the complex of Ganoderma lucidum polysaccharide and jujube seed powder has a synergistic effect in improving sleep duration.

[0068] Furthermore, the number of sleep cycles increased by 52.8%, 54.1%, and 64.2% for the Ganoderma lucidum polysaccharide, jujube seed powder complex, and others, respectively; the sleep improvement rates were 46.1%, 58.9%, and 67.4%, respectively; and the average sleep duration increased by 16.1%, 20.7%, and 16.6%, respectively, indicating that the Ganoderma lucidum polysaccharide and jujube seed powder complex can synergistically improve sleep in zebrafish. Among these, the effect of Example 2 was close to that of melatonin, while the effect of Example 3 was stronger than that of melatonin.

[0069] Experimental Example 3 The experimental method is the same as in Experiment 1. The compound raw materials and results are shown in Table 3. Table 3. Effects of Ganoderma lucidum polysaccharide and hydrolyzed casein peptide complex on sleep in zebrafish.

[0070] Note: The dosages used in Examples 4, 5, and 6 were all 50% effective doses; " / " indicates no numerical value; statistical differences compared to the model group are marked with *. p <0.05,** p <0.01, *** p <0.001.

[0071] Table 3 shows that the sleep improvement rates of zebrafish treated with Ganoderma lucidum polysaccharide and hydrolyzed casein peptide alone were 25.8% and 50.8%, respectively. Based on the EC50 values ​​of Ganoderma lucidum polysaccharide and hydrolyzed casein peptide, the compound ratios of Ganoderma lucidum polysaccharide and hydrolyzed casein peptide were set at 2:1, 2:8, and 8:2, with sleep improvement rates of 56.8%, 62.5%, and 69.5%, respectively, and CI values ​​of 0.39, 0.62, and 0.69, all less than 1. This indicates that when the compound ratio is between 2:8 and 8:2, the complex of Ganoderma lucidum polysaccharide and hydrolyzed casein peptide has a significant synergistic effect in improving sleep improvement rate. In addition, the number of sleep rounds of the Ganoderma lucidum polysaccharide and hydrolyzed casein peptide complex increased by 28.6%, 31.3% and 33.7% respectively compared with the model group, the total sleep duration increased by 50.2%, 58.2% and 65.6% respectively, and the average sleep duration increased by 16.8%, 20.2% and 23.7% respectively, indicating that the complex significantly improved the sleep of zebrafish.

[0072] Experiment Example 4 The experimental method is the same as in Experiment 1. The compound raw materials and results are shown in Table 4. Table 4. Effects of hydrolyzed casein peptide and jujube seed powder complex on sleep in zebrafish.

[0073] Note: The dosages used in Examples 7, 8, and 9 were all 50% effective doses; " / " indicates no numerical value; statistical differences compared to the model group are marked with *. p <0.05,** p <0.01, *** p <0.001.

[0074] Table 4 shows that the number of sleep rounds in zebrafish treated with jujube seed powder and hydrolyzed casein peptide alone increased by 14.2% and 24.7% respectively compared to the model group, demonstrating their potential to improve the number of sleep rounds. Based on the EC50 values ​​of jujube seed powder and hydrolyzed casein peptide, the compound ratios of jujube seed powder and hydrolyzed casein peptide were set to 2:1, 2:8, and 8:2. The number of sleep rounds in the jujube seed powder and hydrolyzed casein peptide complex increased by 34.6%, 35.8%, and 40.7% respectively compared to the model group, with CI values ​​of 0.87, 0.79, and 0.68, all less than 1, indicating a synergistic effect in improving the number of sleep rounds. The sleep improvement rates of the complex were 52.5%, 58.6%, and 61.9%, respectively, with sleep duration increasing by 1.6, 1.6, and 1.7 times, respectively, and average sleep duration increasing by 18.6%, 20.8%, and 23.0%, respectively, indicating that the complex has a synergistic effect in improving zebrafish sleep.

[0075] Experimental Example 5 The experimental method is the same as in Experiment 1. The compound raw materials and results are shown in Table 5. Table 5. Effects of Ganoderma lucidum polysaccharides, hydrolyzed casein peptides, and jujube seed powder complex on sleep in zebrafish.

[0076] Note: The dosages used in each example are 50% of the effective dose; " / " indicates no numerical value; statistical differences compared to the model group are marked with *. p <0.05,** p <0.01, *** p <0.001.

[0077] Table 5 shows that when jujube seed powder, hydrolyzed casein peptide, and Ganoderma lucidum polysaccharide were used alone, the sleep improvement rates were 19.6%, 53.4%, and 25.3%, respectively. The number of sleep rounds, sleep duration, and average sleep duration all increased compared to the model group, indicating that these three ingredients have good potential for improving sleep. Based on the results of Experiments 2-4, jujube seed powder, hydrolyzed casein peptide, and Ganoderma lucidum polysaccharide, when combined in pairs at certain ratios, showed synergistic effects in improving sleep duration, sleep improvement rate, and number of sleep rounds. To obtain scientific and reliable experimental data while shortening the research and development cycle, following publicly reported and accepted experimental group setting methods, hydrolyzed casein peptide and jujube seed powder (1:2), Ganoderma lucidum polysaccharide and jujube seed powder (1:1), and Ganoderma lucidum polysaccharide and hydrolyzed casein peptide powder (2:1) were mixed in fixed ratios and then used as one ingredient to be combined with another ingredient, significantly reducing the interaction after combination to evaluate the experimental groups.

[0078] The results of pairwise compounding showed that the sleep improvement rates of the mixtures of hydrolyzed casein peptide and jujube seed powder (1:2), Ganoderma lucidum polysaccharide and jujube seed powder (1:1), and Ganoderma lucidum polysaccharide and hydrolyzed casein peptide (2:1) were 45.5%, 56.2%, and 51.9%, respectively, with corresponding CI values ​​of 0.72, 0.48, and 0.81, all less than 1. This indicates that the pairwise complexes of jujube seed powder, Ganoderma lucidum polysaccharide, and hydrolyzed casein peptide have a similar effect on improving sleep. The number of sleep rounds increased by 47.5%, 40.1%, and 33.3%, respectively; sleep duration increased by 89.3%, 65.5%, and 66.4%, respectively; and average sleep duration increased by 28.2%, 17.8%, and 24.7%, respectively. This indicates that the pairwise complexes of jujube seed powder, Ganoderma lucidum polysaccharide, and hydrolyzed casein peptide improve sleep.

[0079] The results of the three compounding methods showed that when hydrolyzed casein peptide and jujube seed powder compounded together (1:2) and Ganoderma lucidum polysaccharide were compounded together at ratios of 9:1 (Example 10) and 1:9 (Example 11), the number of sleep rounds increased by 38.9% and 44.4%, sleep duration increased by 74.2% and 84.6%, average sleep duration increased by 25.3% and 27.6%, and sleep improvement rate was 78.5% and 81.7%, respectively. The corresponding CI values ​​were 0.48 and 0.42, both less than 1, indicating that the complex of jujube seed powder, hydrolyzed casein peptide and Ganoderma lucidum polysaccharide has a synergistic effect in improving sleep rate.

[0080] When the compound powder of Ganoderma lucidum polysaccharide and Ziziphus jujuba seed powder (1:1) and hydrolyzed casein peptide were compounded at ratios of 9:1 (Example 12) and 1:9 (Example 13), the number of sleep rounds increased by 46.3% and 48.8%, the sleep duration increased by 102.8% and 103.2%, the average sleep duration increased by 38.5% and 36.2%, and the sleep improvement rate was 83.4% and 84.9%, respectively. The corresponding CI values ​​were 0.49 and 0.36, both less than 1, indicating that the complex of Ziziphus jujuba seed powder, Ganoderma lucidum polysaccharide and hydrolyzed casein peptide has a synergistic effect in improving sleep rate.

[0081] When the compound powder of Ganoderma lucidum polysaccharide and hydrolyzed casein peptide (2:1) was compounded with jujube seed powder at ratios of 9:1 (Example 14) and 1:9 (Example 15), the sleep improvement rates were 80.5% and 81.8%, respectively, with corresponding CI values ​​of 0.44 and 0.52, both less than 1. This indicates that the complex of jujube seed powder, Ganoderma lucidum polysaccharide, and hydrolyzed casein peptide has a synergistic effect in improving sleep. Furthermore, the number of sleep cycles increased by 43.8% and 45.1%, sleep duration increased by 101.9% and 100.3%, and average sleep duration increased by 40.2% and 37.9%, respectively, indicating that the complex of jujube seed powder, Ganoderma lucidum polysaccharide, and hydrolyzed casein peptide has a synergistic effect in improving sleep.

[0082] Based on the results of this example, it can be concluded that when Ganoderma lucidum polysaccharide, jujube seed powder and hydrolyzed casein peptide are combined in a ratio of 1~9:1~9:1~9, their sleep improvement has a significant synergistic effect. The sleep improvement rate of the three combined is better than that of melatonin, and also significantly better than the corresponding pairwise combinations. Among them, Examples 12~15 are close to melatonin in terms of the number of sleep rounds, sleep duration and average sleep duration.

[0083] Experiment Example 6 (Animal Verification Experiment) The complex of Ganoderma lucidum polysaccharides, jujube seed powder, and hydrolyzed casein peptides has been shown to significantly improve sleep in a zebrafish model. To further determine the effects of the combination of these three ingredients on sleep phenotypes and biochemical indicators, and to provide more reliable data support for subsequent product development, the combined formula and dosage, validated in zebrafish experiments, were converted to mouse intake dosages using the dosage conversion method reported in patent CN113496071A. The sleep-improving effect of the combined formula was then verified strictly according to the standard method of "Methods for Functional Testing and Evaluation of Health Products." The specific results are as follows: Table 6. Effects of Ganoderma lucidum polysaccharide, jujube seed powder, and hydrolyzed casein peptide complex on sleep and related indicators in mice.

[0084] Note: " / " indicates no numerical value; statistical differences compared to the model group are marked with *. p <0.05,** p <0.01, ***p <0.001.

[0085] As shown in Table 6, diazepam, as a positive control, at an appropriate dose, shortened the sleep latency by 38.4%, increased the sleep duration by 145.2%, increased the level of the sleep-inducing neurotransmitter serotonin by 31.5%, increased the γ-aminobutyric acid / glutamate ratio by 14.5%, and decreased the excitatory neurotransmitter glutamate by 17.4%. These results indicate that diazepam can significantly improve the sleep phenotype in mice. When hydrolyzed casein peptides and jujube seed powder (1:2) were combined with Ganoderma lucidum polysaccharides at ratios of 9:1 and 1:9, sleep latency decreased by 17.8% and 22.4%, sleep duration increased by 54.6% and 98.4%, serotonin levels increased by 58.2% and 62.0%, the γ-aminobutyric acid / glutamate ratio increased by 4.8% and 8.1%, and the excitatory neurotransmitter glutamate decreased by 9.5% and 20.6%. These results indicate that the complex of hydrolyzed casein peptides and jujube seed powder with Ganoderma lucidum polysaccharides can improve sleep in mice.

[0086] When jujube seed powder and Ganoderma lucidum polysaccharide compound powder (1:1) and hydrolyzed casein peptides were compounded at ratios of 9:1 and 1:9, the sleep latency decreased by 29.7% and 31.7%, the sleep duration increased by 114.9% and 122.0%, the serotonin level increased by 67.9% and 71.7%, the γ-aminobutyric acid / glutamate ratio increased by 12.9% and 16.1%, and the excitatory neurotransmitter glutamate decreased by 24.9% and 29.3%. These results indicate that jujube seed powder and Ganoderma lucidum polysaccharide compound powder combined with hydrolyzed casein peptides can improve sleep in mice.

[0087] When hydrolyzed casein peptide and Ganoderma lucidum polysaccharide compound powder (1:2) was compounded with jujube seed powder at ratios of 9:1 and 1:9, the sleep latency decreased by 29.5% and 31.6%, the sleep duration increased by 119.3% and 118.0%, the 5-hydroxytryptamine level increased by 58.5% and 61.7%, the γ-aminobutyric acid / glutamate ratio increased by 8.1% and 11.3%, and the excitatory neurotransmitter glutamate decreased by 27.3% and 30.3%. These results indicate that the complex of hydrolyzed casein peptide and Ganoderma lucidum polysaccharide compound powder (1:2) with jujube seed powder can improve sleep in mice.

[0088] Based on the results of this case, it can be concluded that the compound powder of Ganoderma lucidum polysaccharide, jujube seed powder and hydrolyzed casein peptide can significantly shorten sleep latency and increase sleep duration by increasing 5-hydroxytryptamine levels and the γ-aminobutyric acid / glutamate ratio, and decreasing the level of the excitatory neurotransmitter glutamate.

[0089] Thus, the composition of this application has been proven to have a significant effect on improving sleep by the standard method of "Methods for Functional Testing and Evaluation of Health Foods".

[0090] Experiment Example 7 (Trial Results for a Population) A total of 56 volunteers were recruited through our unit's work group. The Pittsburgh Sleep Quality Index (PSQI) and Polysomnography (PSG) were used to assess the sleep quality of the recruits. Further screening was conducted, and volunteers scoring 7-10 (mild sleep disorder) and 10-15 (moderate sleep disorder) were selected as the test group (a total of 28 volunteers). No control group was included in the experiment. The experiment only tracked the sleep improvement effect of the pilot-scale production beverage (formula from Example 14, containing 180 mg of Ganoderma lucidum polysaccharide, 30 mg of jujube seed powder, and 90 mg of hydrolyzed casein peptide dissolved in 25 ml of purified water, sterilized and packaged, and administered to volunteers 10-30 minutes before bedtime) for 7 consecutive days. A total of 26 valid questionnaires were collected. The statistical results are as follows: Figure 1 and Figure 2 As shown.

[0091] Depend on Figure 1 It was found that after taking a compound beverage of Ganoderma lucidum polysaccharides, jujube seed powder, and hydrolyzed casein peptides for 7 days, 95% of the participants experienced a reduction in sleep duration. Figure 1 In the middle (A) group, 84% of people experienced an increase in total sleep time. Figure 1 In China (B), 79% of the population experienced improved sleep quality. Figure 1 (C) 89% of people experienced improved mental state after waking up. Figure 1 (D) Figure 2 It can be seen that by plotting the 7-day data of the test group into a curve, it was found that as the duration of use increased, the time it took for the group to fall asleep decreased. Figure 2 (A) ), Total sleep time ( Figure 2 (B) ), sleep quality ( Figure 2 (C) and mental state after waking up ( Figure 2 All three (D) showed continuous improvement. These results indicate that the compound beverage of Ganoderma lucidum polysaccharide, jujube seed powder and hydrolyzed casein peptide can significantly improve mild to moderate sleep disorders, and the improvement effect is good and lasting.

[0092] Comparative Example Table 7 shows the comparative proportions of raw materials and results. All raw materials were obtained commercially. The specific results are as follows: Comparative Example 1 The components in this formula are (parts by weight): 1 part licorice powder and 2 parts jujube powder.

[0093] Comparative Example 2 The components in this recipe are (parts by weight): 1 part licorice powder and 3 parts wheat flour.

[0094] Comparative Example 3 The components in this recipe are (by weight): 2 parts jujube powder and 3 parts wheat flour.

[0095] Comparative Example 4 The components in this recipe are (parts by weight): 1 part licorice powder, 2 parts jujube powder and 3 parts wheat flour.

[0096] The zebrafish experiment was conducted using the same method as in Experimental Example 1, and the results are as follows: Table 7. Effects of comparative studies on zebrafish sleep

[0097] Table 7 shows that the sleep duration of the licorice powder, wheat flour, and jujube powder treatment groups increased by 11.5%, 13.0%, and 11.1% respectively compared to the model group, with sleep improvement rates of 11.8%, 15.4%, and 18.2%, respectively. This indicates that these ingredients, when used alone, have sleep-improving effects. However, regardless of whether these ingredients were combined in pairs or in all three combinations, although the sleep improvement rate increased significantly, the corresponding CI values ​​were 1.61, 1.52, 1.42, and 1.28, all greater than 1. This proves that the various combinations of these three ingredients do not have a synergistic effect, and also indicates that not all single ingredients with sleep-improving effects have a synergistic effect when combined.

[0098] The embodiments described above are merely illustrative of several implementation methods of this application, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the protection scope of this application. Therefore, the protection scope of this patent application should be determined by the appended claims.

Claims

1. A composition for improving sleep, characterized in that, By weight, its components are: 0.5-9 parts of Ganoderma lucidum polysaccharide; 0.3-9 parts of hydrolyzed casein peptide; and 0.5-9 parts of jujube seed powder.

2. The composition according to claim 1, characterized in that, The Ganoderma lucidum polysaccharide contains more than 5% by mass; the hydrolyzed casein peptides contain at least 80% protein hydrolysates with a relative molecular mass of less than 1000 Da and at least 90% oligopeptides with a relative molecular mass of less than 3000 Da; and the jujube seed powder contains at least 1% total jujuboside by mass.

3. The composition according to claim 1, characterized in that, The hydrolyzed casein peptide and jujube seed powder are compounded at a weight ratio of 1:2, and then compounded with Ganoderma lucidum polysaccharide at a weight ratio of 9:1 to 1:9 based on the total weight of the two.

4. The composition according to claim 1, characterized in that, The Ganoderma lucidum polysaccharide and jujube seed powder are compounded at a weight ratio of 1:11, and then compounded with hydrolyzed casein peptide at a weight ratio of 9:1 to 1:

9.

5. The composition according to claim 1, characterized in that, The Ganoderma lucidum polysaccharide and hydrolyzed casein peptide are compounded at a weight ratio of 2:1, and then compounded with the hydrolyzed casein peptide at a weight ratio of 9:1 to 1:9 of the total weight of the two and the jujube seed powder.

6. A composition for improving sleep, characterized in that, The composition is a compound of Ganoderma lucidum polysaccharide and jujube seed powder in a weight ratio of 1:9 to 9:

1.

7. Use of the composition according to any one of claims 1 to 6 in the preparation of a sleep-improving product.

8. The application according to claim 7, characterized in that, The sleep-improving products mentioned are medicines or health foods.

9. The application according to claim 8, characterized in that, The health food products mentioned are solid beverages, compressed candies, or oral liquids.

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