A composition and its use in the preparation of sleep-improving pharmaceuticals and health foods

Abstract

The application discloses a composition and application thereof in preparing sleep-improving medicines and health foods. Components of the composition include ganoderma lucidum polysaccharide, hydrolyzed casein peptide and spina date seed powder, and the components can be any two components compounded in a certain ratio, or the three components compounded in a certain ratio. In the three-component compounded composition, the components are ganoderma lucidum polysaccharide 0.5-9 parts by weight, hydrolyzed casein peptide 0.3-9 parts by weight, and spina date seed powder 0.5-9 parts by weight. In the two-component compounded composition, the ganoderma lucidum polysaccharide and the spina date seed powder are compounded in a weight ratio of 1:9-9:1. It is verified that the composition has a significant synergistic effect on the number of sleep cycles, the total sleep time and the average sleep time, and the composition is simple in components, safe in source, easy to prepare and easy to industrial production.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biological medicine and health food, and particularly relates to a composition and application thereof in preparing sleep-improving medicine and health food. BACKGROUND

[0002] With the acceleration of modern life pace, sleep problems are increasingly prominent, mainly manifested as late sleep time, sleep difficulty, insufficient sleep time, poor sleep quality and the like. Sleep problems not only affect daily life and study, but also can induce various health problems.

[0003] In recent years, numerous patents and literatures report natural small molecule compounds, traditional Chinese medicine compositions, health food compositions, natural food raw materials and the like for improving sleep. For example, patent document CN 105902536A discloses application of loganin in preparing sleep-improving medicine or health food. Animal experiments prove that loganin can prolong the total sleep time of mice and dose-dependently increase the sleep amount. Patent document CN 102309565A discloses a traditional Chinese medicine composition for improving sleep and a preparation method thereof. The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 150-250 parts of Chinese date kernel extract, 100-150 parts of Platycladus orientalis kernel extract, 100-150 parts of prepared Radix Polygoni Multiflori extract, 100-120 parts of Herba Epimedii extract, 100-150 parts of Jujube extract, 100-150 parts of Acanthopanax senticosus extract and 1-5 parts of vitamin B1.

[0004] However, single component has limited improvement effect, chemical medicine has many side effects, and the composition has problems such as complex components, complicated preparation or unknown compatibility interaction. SUMMARY

[0005] The present application provides a composition and application thereof in preparing sleep-improving medicine and health food. The composition has significant synergistic effect on sleep cycle number, total sleep time and average sleep time, and has simple components, safe source and easy preparation, and is easy for industrial production.

[0006] The present application constructs an insomnia model by inducing sleep behavior disorder of zebrafish larvae with pentylenetetrazole, and evaluates the improvement effect of two or more combinations of hydrolyzed casein peptide, Ganoderma lucidum polysaccharide, Chinese date kernel powder, longan, asparagus and lily on sleep phenotype (sleep improvement rate, sleep cycle number, sleep time and average sleep time) by using a behavior analysis instrument, and compares the sleep-improving effect with that of melatonin, a recognized sleep-improving hormone.

[0007] And through the combination index method quantitative analysis of the composition of the synergistic effect on sleep, finally, with diazepam as a positive modeling drug, through the mouse-based animal test to verify the synergistic formula to improve the effect of sleep behavior, and its influence on sleep neurotransmitter and hormone levels, in order to verify the sleep improvement effect. Finally, based on the crowd trial data, a composition with significant synergistic effect on sleep is developed.

[0008] The components of the composition include ganoderma lucidum polysaccharide, hydrolyzed casein peptide and spina date seed powder, which can be any two components compounded in a certain ratio, or three components compounded in a certain ratio. The sleep improvement rate of the composition under a certain ratio is better than that of melatonin, and the sleep round times, sleep duration and average sleep duration are close to those of melatonin.

[0009] The polysaccharide, triterpenoid compounds and other components in ganoderma lucidum can improve sleep through multiple mechanisms. Among them, ganoderma lucidum polysaccharide can regulate the nervous system, promote the pineal gland to secrete melatonin, shorten the sleep latency and increase the sleep duration. Hydrolyzed casein peptide can increase the content of 5-hydroxytryptamine, the precursor of melatonin in the brain, and significantly shorten the sleep latency. Hydrolyzed casein peptide can also bind to γ-aminobutyric acid receptors, enhance the effect of inhibitory neurotransmitter γ-aminobutyric acid, and at the same time reduce the level of excitatory neurotransmitter glutamate, inhibit the excitation of the nervous system, and promote deep sleep. As the core component of traditional Chinese medicine spina date seed, spina date seed glycoside in spina date seed powder can improve sleep through multiple pathways such as regulating neurotransmitters, inhibiting the excessive activation of hypothalamic-pituitary-adrenal axis, anti-inflammatory, protecting neurons and promoting neuron health. The sedative and hypnotic effect of spina date seed glycoside is not only suitable for insomnia patients, but also has auxiliary treatment effect on diseases such as neurasthenia and anxiety.

[0010] According to the standard method of "Health Product Function Test and Evaluation Method", the sleep improvement effect of the composition of the present application is verified, and the results show that the compound powder of ganoderma lucidum polysaccharide, spina date seed powder and hydrolyzed casein peptide can significantly shorten the sleep latency, increase the sleep duration, improve the sleep of mice by increasing the level of 5-hydroxytryptamine and the ratio of γ-aminobutyric acid / glutamic acid, and reducing the level of excitatory neurotransmitter glutamic acid, and is proved to have significant sleep improvement effect.

[0011] The sleep improvement effect of the pilot drink with the composition of the present application as the core formula is also verified by recruiting volunteers, and the results show that after taking the compound drink of ganoderma lucidum polysaccharide, spina date seed powder and hydrolyzed casein peptide for 7 days, 95% of the crowd's sleep time is shortened, 84% of the crowd's total sleep time is prolonged, 79% of the crowd's sleep quality is improved, and 89% of the crowd's mental state after waking up is improved. By drawing the data of the test crowd for 7 days into a curve graph, it is found that with the extension of the taking time, the sleep time, total sleep time, sleep quality and mental state after waking up of the crowd are continuously improved.

[0012] Based on the above research results, the application first provides a composition for improving sleep, which is a compound product of ganoderma polysaccharide, hydrolyzed casein peptide and spina date seed powder, and the components are as follows in terms of weight parts: ganoderma polysaccharide 0.5-9 parts; hydrolyzed casein peptide 0.3-9 parts; and spina date seed powder 0.5-9 parts.

[0013] The sources of the components in the composition can be obtained by purchase, and the components obtained by purchase are all in powder form, usually in the form of water extract of the corresponding raw material, and the preparation method can be conventional mixing, for example, uniformly mixing the powdery raw materials, for the composition of three components, two components can be compounded according to a specific ratio and then compounded with the third component according to a specific ratio.

[0014] The components in the composition can also be prepared by conventional processing methods in the art: the raw materials corresponding to the components, ganoderma, casein and spina date seed, are obtained by one-time extraction, concentration / drying, etc. with a common solvent, and then compounded; or the raw materials are weighed according to a certain proportion and physically mixed, and then obtained by one-time extraction, concentration / drying, etc. with a common solvent.

[0015] The preparation method of ganoderma polysaccharide is as follows: the qualified ganoderma is crushed into ultra-fine powder, water or low proportion of ethanol is used as the solvent, then the appropriate solid-liquid ratio (1:10-1:30) is used for extraction (100-150 ℃, 2 hours) in a multifunctional extraction tank, plate frame filtration, membrane concentration, and freeze-drying to obtain a powder, which is used as a compounding raw material in the following examples.

[0016] The preparation method of spina date seed powder is as follows: the qualified spina date seed is crushed into ultra-fine powder, water or low proportion of ethanol is used as the solvent, then the appropriate solid-liquid ratio (1:10-1:20) is used for cyclic extraction (50 ℃, 1 hour) in a multifunctional extraction tank, gauze filtration, membrane concentration, and freeze-drying to obtain a powder, which is used as a compounding raw material in the following examples.

[0017] The preparation method of hydrolyzed casein peptide is as follows: water is used as the solvent, the casein is added with 1-10% of complex enzymes (neutral protease, bromelain and flavor enzyme, etc. according to the need) according to the appropriate solid-liquid ratio (usually 1:10-1:100), and then enzymolysis is carried out in an enzymolysis tank for 4-6 hours (pH 5.0-8.5, 45-65 ℃), high-temperature enzyme inactivation (90 ℃, 15 minutes), and then centrifugal separation, concentration and spray drying are carried out.

[0018] No matter whether it is obtained by purchase or self-preparation, the components preferably meet the following conditions:

[0019] The content of ganoderma lucidum polysaccharide in the ganoderma lucidum polysaccharide is greater than 5%; the content of protein hydrolysate with a relative molecular weight of less than 1000 Da in the hydrolyzed casein peptide is not less than 80%, and the content of oligopeptide (molecular weight less than 3000 Da) is not less than 90%; and the content of total jujube glycosides in the jujube seed powder is not less than 1%.

[0020] A preferred three-component compounded composition, the hydrolyzed casein peptide and the jujube seed powder are compounded at a weight ratio of 1:2, and then the total weight of the two is compounded with the ganoderma lucidum polysaccharide at a weight ratio of 9:1 to 1:9.

[0021] Another preferred three-component compounded composition, the ganoderma lucidum polysaccharide and the jujube seed powder are compounded at a weight ratio of 1:11, and then the total weight of the two is compounded with the hydrolyzed casein peptide at a weight ratio of 9:1 to 1:9.

[0022] Still another preferred three-component compounded composition, the ganoderma lucidum polysaccharide and the hydrolyzed casein peptide are compounded at a weight ratio of 2:1, and then the total weight of the two is compounded with the jujube seed powder at a weight ratio of 9:1 to 1:9.

[0023] The composition of the present application can also be a compound of any two components:

[0024] An optional two-by-two compounding scheme, the ganoderma lucidum polysaccharide and the jujube seed powder are compounded at a weight ratio of 1:9 to 9:1. Further, the ganoderma lucidum polysaccharide and the jujube seed powder are compounded at a weight ratio of 1:9, 1:1 or 9:1. When the ganoderma lucidum polysaccharide and the jujube seed powder are compounded at a weight ratio of 1:9, the improvement effect is close to that of melatonin, and when the ganoderma lucidum polysaccharide and the jujube seed powder are compounded at a weight ratio of 9:1, the improvement effect is better than that of melatonin.

[0025] Another optional two-by-two compounding scheme, the ganoderma lucidum polysaccharide and the hydrolyzed casein peptide are compounded at a weight ratio of 1:4 to 4:1. Further, the ganoderma lucidum polysaccharide and the hydrolyzed casein peptide are compounded at a weight ratio of 1:4, 2:1 or 4:1.

[0026] Still another optional two-by-two compounding scheme, the jujube seed powder and the hydrolyzed casein peptide are compounded at a weight ratio of 1:4 to 4:1. Further, the jujube seed powder and the hydrolyzed casein peptide are compounded at a weight ratio of 1:4, 2:1 or 4:1.

[0027] Based on the efficacy of the composition verified in the present application, the present application also provides an application of the composition in preparing a sleep improvement product.

[0028] Optionally, the sleep improvement includes but is not limited to improving sleep improvement rate, increasing sleep round number, sleep duration and average sleep duration.

[0029] Optionally, the sleep improvement product is a drug or a health food.

[0030] For preparing a health food, the composition can be prepared with food acceptable adjuvants such as solvents, binders, fillers, sugars and sweeteners, etc. according to conventional food preparation methods. The type of the health food can be any one of tablets, capsules, oral liquids, solid beverages, pressed candy, pills, granules, decoctions, pastes, lotions, dripping pills or syrups or tea bags. Optionally, the health food is a solid beverage, a pressed candy or an oral liquid.

[0031] Further preferably, when the composition is composed of 9 parts of Ganoderma lucidum polysaccharide and hydrolyzed casein peptide compound powder (2:1) and 1 part of spina date seed powder by weight; in the oral liquid: Ganoderma lucidum polysaccharide 7.6 mg / mL, spina date seed powder 1.2 mg / mL, and hydrolyzed casein peptide 3.6 mg / mL.

[0032] For preparing a drug, the composition can be prepared with pharmaceutically acceptable adjuvants according to conventional drug preparation methods, which include buffers, emulsifiers, suspending agents, decomposing agents, disintegrating agents, dispersing agents, binders, excipients, stabilizers, chelating agents, diluents, gelling agents, preservatives, wetting agents, lubricants, absorption delaying agents, liposomes, etc. which can be selected according to different needs. The dosage form of the drug includes granules, tablets, capsules, granules, granules, oral liquids, dripping pills, micro-pellets or injections, etc.

[0033] In the above health food and drug, the addition amount and addition ratio of the composition and related adjuvants of the present application can be adjusted according to actual needs.

[0034] Compared with existing products, the present application has the following beneficial effects:

[0035] The composition of the present application has been proved to have a significant sleep improvement effect. Ganoderma lucidum polysaccharide, hydrolyzed casein peptide and spina date seed powder play a synergistic effect, and have a significant improvement effect on sleep round number, total sleep duration and average sleep duration, close to or better than the positive control. The composition is simple in components, safe in source and easy to prepare and industrial production. BRIEF DESCRIPTION OF DRAWINGS

[0036] Figure 1Figures showing the results of sleep onset time (A), total sleep time (B), sleep quality (C), and mental state after waking up (D) for the volunteer group in Test Example 7.

[0037] Figure 2 Figures showing the improvement curves of 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 taking the composition continuously for 7 days. DETAILED DESCRIPTION

[0038] The technical solutions of the present application will be described in detail below with specific examples. Obviously, the described examples are only a part of the examples of the present application, but not all the examples. Based on the examples in the present application, all other examples obtained by those skilled in the art without creative work fall within the scope of protection of the present application.

[0039] 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 the present application belongs. The terminology used in the description of the present application herein is only for the purpose of describing specific embodiments of the present application and is not intended to limit the present application.

[0040] Ganoderma lucidum polysaccharide (extract, content of Ganoderma lucidum polysaccharide is greater than 5%), hydrolyzed casein peptide (protein hydrolysate with relative molecular mass less than 1000 Da accounts for not less than 80%, oligopeptide (molecular weight less than 3000 Da) not less than 90%), and spina date seed powder (total spina date glycoside content not less than 1%) are all purchased from the market.

[0041] Example 1

[0042] The composition of the present example contains, by weight, 1 part of Ganoderma lucidum polysaccharide and 1 part of spina date seed powder.

[0043] Example 2

[0044] The composition of the present example contains, by weight, 1 part of Ganoderma lucidum polysaccharide and 9 parts of spina date seed powder.

[0045] Example 3

[0046] The composition of the present example contains, by weight, 9 parts of Ganoderma lucidum polysaccharide and 1 part of spina date seed powder.

[0047] Example 4

[0048] The composition of the present example contains, by weight, 2 parts of Ganoderma lucidum polysaccharide and 1 part of hydrolyzed casein peptide.

[0049] Example 5

[0050] The composition of this example has the following components by weight parts: 2 parts of Ganoderma lucidum polysaccharide, 8 parts of hydrolyzed casein peptide.

[0051] Example 6

[0052] The composition of this example has the following components by weight parts: 8 parts of Ganoderma lucidum polysaccharide, 2 parts of hydrolyzed casein peptide.

[0053] Example 7

[0054] The composition of this example has the following components by weight parts: 2 parts of Zizyphus jujuba Mill. powder, 1 part of hydrolyzed casein peptide.

[0055] Example 8

[0056] The composition of this example has the following components by weight parts: 2 parts of Zizyphus jujuba Mill. powder, 8 parts of hydrolyzed casein peptide.

[0057] Example 9

[0058] The composition of this example has the following components by weight parts: 8 parts of Zizyphus jujuba Mill. powder, 2 parts of hydrolyzed casein peptide.

[0059] Example 10

[0060] The composition of this example has the following components by weight parts: 9 parts of hydrolyzed casein peptide and Zizyphus jujuba Mill. powder compound powder (1:2), 1 part of Ganoderma lucidum polysaccharide.

[0061] Example 11

[0062] The composition of this example has the following components by weight parts: 1 part of hydrolyzed casein peptide and Zizyphus jujuba Mill. powder compound powder (1:2), 9 parts of Ganoderma lucidum polysaccharide.

[0063] Example 12

[0064] The composition of this example has the following components by weight parts: 9 parts of Ganoderma lucidum polysaccharide and Zizyphus jujuba Mill. powder compound powder (1:1), 1 part of hydrolyzed casein peptide.

[0065] Example 13

[0066] The composition of this example has the following components by weight parts: 1 part of Ganoderma lucidum polysaccharide and Zizyphus jujuba Mill. powder compound powder (1:1), 9 parts of hydrolyzed casein peptide.

[0067] Example 14

[0068] The composition of this example has the following components by weight parts: 9 parts of Ganoderma lucidum polysaccharide and hydrolyzed casein peptide compound powder (2:1), 1 part of Zizyphus jujuba Mill. powder.

[0069] Example 15

[0070] The composition of the present embodiment, in parts by weight, has the following components: 1 part of Ganoderma lucidum polysaccharide and hydrolyzed casein peptide compound powder (2:1), and 9 parts of Zizyphus jujuba Mill. powder.

[0071] Experimental Example 1 (raw material screening)

[0072] Select 6 dpf wild-type AB strain zebrafish to induce insomnia model with pentylenetetrazole. Use melatonin as a positive control, and set the melatonin administration concentration to 100 μM. Before the start of the behavior experiment, add pentylenetetrazole solution to each group of cultured water, and timely drop melatonin stock solution and treatment group stock solution respectively to make the dose meet the experimental set dose. The compound raw materials are shown in Table 1.

[0073] Among them, Zizyphus jujuba Mill. powder (EC50 is 210 μg·mL -1 ), hydrolyzed casein peptide (EC50 is 114.5 μg·mL -1 ) and Ganoderma lucidum polysaccharide (extract, EC50 is 164.6 μg·mL -1 ) are set according to their respective EC50 values; the dose of Asparagus officinalis L. powder is set to 750 μg·mL -1 , the dose of Longan powder is set to 550 μg·mL -1 , and the dose of Lily powder is set to 600 μg·mL -1 .

[0074] Record the zebrafish trajectory for 1 h using a behavior analysis instrument, where 0.5 cm / s or less is slow motion, 0.5-2 cm / s is medium motion, and more than 2 cm / s is high-speed motion. The total number of sleep cycles within 1 h is recorded as the total number of sleep cycles. Slow motion duration greater than or equal to 6 s is recorded as one successful sleep. Sleep duration: total sleep time within 1 h. Average sleep duration: average sleep duration within 1 h.

[0075] The sleep improvement rate is as follows:

[0076] Sleep improvement rate = 100 x (total high-speed motion distance of the model group - total high-speed motion distance of the treatment group) / total high-speed motion distance of the model group.

[0077] The specific results are as follows:

[0078] Table 1 Screening of sleep improvement compound formula raw materials based on zebrafish insomnia model

[0079]

[0080] From Table 1, the model group sleep round number is 15.6, sleep duration is 274.8 s, and the average sleep duration is 17.6 s, which are significantly lower than those of the normal group. The experimental results are consistent with the insomnia phenotype of zebrafish induced by pentylenetetrazole reported in many literatures. At the same time, the sleep improvement rate of the group added with the modeling drug and melatonin treatment is 76.2%, the sleep round number is 25.6, the sleep duration is 728.5 s, and the average sleep duration is 28.4 s, indicating that melatonin can reverse the insomnia phenotype induced by pentylenetetrazole. Therefore, the zebrafish insomnia model induced by pentylenetetrazole is successfully constructed.

[0081] Based on the established zebrafish insomnia model, the improvement effects of six food raw materials, including ganoderma lucidum polysaccharide, spina date seed powder, hydrolyzed casein peptide, asparagus officinalis powder, longan powder and lily powder, on the insomnia phenotype were evaluated.

[0082] In terms of sleep improvement rate, the improvement effects are in the order of hydrolyzed casein peptide > ganoderma lucidum polysaccharide > spina date seed powder > longan powder > asparagus officinalis powder > lily powder.

[0083] In terms of sleep round number, the improvement effects are in the order of hydrolyzed casein peptide > spina date seed powder > longan powder > ganoderma lucidum polysaccharide > lily powder > asparagus officinalis powder.

[0084] In terms of sleep duration, the improvement effects are in the order of spina date seed powder > ganoderma lucidum polysaccharide > hydrolyzed casein peptide > longan powder > lily powder > asparagus officinalis powder.

[0085] From the research results in Table 1, it can be seen that hydrolyzed casein peptide and ganoderma lucidum polysaccharide can both improve the sleep improvement rate, and the combination of the two raw materials may have synergistic effect. Hydrolyzed casein peptide and spina date seed powder can both significantly improve the sleep round number, and the combination of the two raw materials is expected to have synergistic effect. Spina date seed powder and ganoderma lucidum polysaccharide can both significantly prolong the sleep duration, and the combination of the two raw materials may have synergistic effect.

[0086] Considering the improvement effects and contribution rates of the six food raw materials on the sleep phenotype, ganoderma lucidum polysaccharide, spina date seed powder and hydrolyzed casein peptide are selected as the core raw materials of the sleep improvement compound formula. Finally, on the basis of the combination of the two raw materials and the experimental verification of significant synergism, the third raw material is combined to verify the synergism of the three raw materials. The present application aims to develop a sleep improvement compound formula with synergistic effect. The subsequent examples will focus on these speculations.

[0087] Experimental Example 2

[0088] The experimental method is the same as that in Experimental Example 1, and the combination of raw materials and results are shown in Table 2:

[0089] Table 2 Effect of Ganoderma Lucidum Polysaccharide and Spina Date Seed Powder Compound on Zebrafish Sleep

[0090]

[0091] Note: The dosage of Example 1, Example 2 and Example 3 is 50% effective dose; " / " means no value; the statistical difference compared with the model group is marked as * p <0.05,** p <0.01,*** p <0.001.

[0092] As shown in Table 2, the sleep duration of zebrafish treated with Ganoderma lucidum polysaccharide and spina date seed powder alone increased by 30.5% and 41.7% respectively compared with the model group. According to the EC50 value (half effective concentration) of raw materials in the screening stage, the compound ratio of Ganoderma lucidum polysaccharide and spina date seed powder was set as 1:1, 1:9 and 9:1, and the corresponding CI values were 0.79, 0.69 and 0.62 respectively, all less than 1, indicating that when the compound ratio was 1:9~9:1, the Ganoderma lucidum polysaccharide and spina date seed powder compound had synergistic effect on improving sleep duration.

[0093] In addition, the sleep round times of Ganoderma lucidum polysaccharide and spina date seed powder compound increased by 52.8%, 54.1% and 64.2% 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 Ganoderma lucidum polysaccharide and spina date seed powder compound could synergistically improve zebrafish sleep. Among them, the effect of Example 2 was close to melatonin, and the effect of Example 3 was stronger than melatonin.

[0094] Experimental Example 3

[0095] The experimental method is the same as that of Experimental Example 1, and the compound raw materials and results are shown in Table 3:

[0096] Table 3 Effect of Ganoderma lucidum polysaccharide and hydrolyzed casein peptide compound on sleep of zebrafish

[0097]

[0098] Note: The dosage of Example 4, Example 5 and Example 6 is 50% effective dose; " / " means no value; the statistical difference compared with the model group is marked as * p <0.05,** p <0.01,*** p <0.001.

[0099] From Table 3, the sleep improvement rates of zebrafish treated with Ganoderma lucidum polysaccharide and hydrolyzed casein peptide alone were 25.8% and 50.8%, respectively. According to the EC50values of Ganoderma lucidum polysaccharide and hydrolyzed casein peptide, the compound ratios of Ganoderma lucidum polysaccharide and hydrolyzed casein peptide were set to 2:1, 2:8 and 8:2, and the sleep improvement rates were 56.8%, 62.5% and 69.5%, respectively, and the CI values were 0.39, 0.62 and 0.69, respectively, all less than 1, indicating that when the compound ratio was 2:8~8:2, the Ganoderma lucidum polysaccharide and hydrolyzed casein peptide compound had significant synergistic effect on improving sleep improvement rate. In addition, the sleep round times of the Ganoderma lucidum polysaccharide and hydrolyzed casein peptide compound were increased by 28.6%, 31.3% and 33.7% compared with the model group, the total sleep time was increased by 50.2%, 58.2% and 65.6%, and the average sleep time was increased by 16.8%, 20.2% and 23.7%, respectively, indicating that the compound significantly improved zebrafish sleep.

[0100] Experimental Example 4

[0101] The experimental method was the same as that in Experimental Example 1, and the compound raw materials and results are shown in Table 4:

[0102] Table 4 Effect of hydrolyzed casein peptide and spina date kernel powder compound on zebrafish sleep

[0103]

[0104] Note: The dosage of Example 7, Example 8 and Example 9 was 50% effective dose; " / " represents no numerical value; the statistical difference compared with the model group is denoted as * p <0.05, p <0.01, p <0.001.

[0105] From Table 4, the sleep round times of zebrafish treated with spina date kernel powder and hydrolyzed casein peptide alone were increased by 14.2% and 24.7% compared with the model group, showing the potential to improve sleep round times. According to the EC50values of spina date kernel powder and hydrolyzed casein peptide, the compound ratios of spina date kernel powder and hydrolyzed casein peptide were set to 2:1, 2:8 and 8:2, and the sleep round times of the spina date kernel powder and hydrolyzed casein peptide compound were increased by 34.6%, 35.8% and 40.7% compared with the model group, and the CI values were 0.87, 0.79 and 0.68, respectively, all less than 1, showing synergistic effect on improving sleep round times. The sleep improvement rates of the compound were 52.5%, 58.6% and 61.9%, respectively, the sleep time was increased by 1.6, 1.6 and 1.7 times, respectively, and the average sleep time was increased by 18.6%, 20.8% and 23.0%, respectively, indicating that the compound had synergistic effect on improving zebrafish sleep.

[0106] Experimental Example 5

[0107] The experimental method is the same as that in Experimental Example 1, and the compounded raw materials and results are shown in Table 5:

[0108] Table 5 Effect of Ganoderma lucidum polysaccharide, hydrolyzed casein peptide and Semen Ziziphi Spinosae powder complex on sleep of zebrafish

[0109]

[0110] Note: The dosage of each example is 50% effective dose; " / " indicates no value; statistical difference compared with the model group is marked as * p <0.05,** p <0.01,*** p <0.001.

[0111] As shown in Table 5, when Semen Ziziphi Spinosae powder, hydrolyzed casein peptide and Ganoderma lucidum polysaccharide are used alone, the sleep improvement rates are 19.6%, 53.4% and 25.3% respectively, and the sleep round times, sleep duration and average sleep duration are all increased compared with the model group, indicating that these three raw materials have good potential to improve sleep. According to the results of Experimental Examples 2-4, Semen Ziziphi Spinosae powder, hydrolyzed casein peptide and Ganoderma lucidum polysaccharide show synergistic effect in improving sleep duration, sleep improvement rate and sleep round times when they are compounded in a certain ratio. In order to shorten the research and development cycle and obtain scientific and reliable experimental data, according to the experimental group setting method reported and recognized in the public literature, the hydrolyzed casein peptide and Semen Ziziphi Spinosae powder compound powder (1:2), the Ganoderma lucidum polysaccharide and Semen Ziziphi Spinosae powder compound powder (1:1) and the Ganoderma lucidum polysaccharide and hydrolyzed casein peptide compound powder (2:1) are mixed in a fixed ratio, and then they are compounded with another raw material as a kind of raw material, so as to significantly reduce the evaluation of experimental groups after compounding.

[0112] The results of two-by-two compounding show that after mixing the hydrolyzed casein peptide and Semen Ziziphi Spinosae powder compound powder (1:2), the Ganoderma lucidum polysaccharide and Semen Ziziphi Spinosae powder compound powder (1:1) and the Ganoderma lucidum polysaccharide and hydrolyzed casein peptide compound powder (2:1), the sleep improvement rates are 45.5%, 56.2% and 51.9% respectively, and the corresponding CI values are 0.72, 0.48 and 0.81 respectively, all less than 1, indicating that the sleep improvement rates of the two-by-two compound of Semen Ziziphi Spinosae powder, Ganoderma lucidum polysaccharide and hydrolyzed casein peptide have the same effect. The sleep round times are increased by 47.5%, 40.1% and 33.3% respectively, the sleep duration is increased by 89.3%, 65.5% and 66.4% respectively, and the average sleep duration is increased by 28.2%, 17.8% and 24.7% respectively, indicating that the two-by-two compound of Semen Ziziphi Spinosae powder, Ganoderma lucidum polysaccharide and hydrolyzed casein peptide improves sleep.

[0113] The results of the three combinations showed that after the hydrolyzed casein peptide and the Chinese wingnut powder (1:2) were compounded with ganoderma lucidum polysaccharide at 9:1 (Example 10) and 1:9 (Example 11), the sleep cycle times increased by 38.9% and 44.4%, the sleep duration increased by 74.2% and 84.6%, the average sleep duration increased by 25.3% and 27.6%, the sleep improvement rate was 78.5% and 81.7%, and the corresponding CI values were 0.48 and 0.42, both of which were less than 1, indicating that the sleep improvement rate of the compound of Chinese wingnut powder, hydrolyzed casein peptide and ganoderma lucidum polysaccharide had synergism.

[0114] After the ganoderma lucidum polysaccharide and the Chinese wingnut powder (1:1) were compounded with the hydrolyzed casein peptide at 9:1 (Example 12) and 1:9 (Example 13), the sleep cycle times 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%, the sleep improvement rate was 83.4% and 84.9%, and the corresponding CI values were 0.49 and 0.36, both of which were less than 1, indicating that the sleep improvement rate of the compound of Chinese wingnut powder, ganoderma lucidum polysaccharide and hydrolyzed casein peptide had synergism.

[0115] After the ganoderma lucidum polysaccharide and the hydrolyzed casein peptide compound powder (2:1) were compounded with the Chinese wingnut powder at 9:1 (Example 14) and 1:9 (Example 15), the sleep improvement rate was 80.5% and 81.8%, and the corresponding CI values were 0.44 and 0.52, both of which were less than 1, indicating that the sleep improvement rate of the compound of Chinese wingnut powder, ganoderma lucidum polysaccharide and hydrolyzed casein peptide had synergism. The sleep cycle times increased by 43.8% and 45.1%, the sleep duration increased by 101.9% and 100.3%, the average sleep duration increased by 40.2% and 37.9%, indicating that the compound of Chinese wingnut powder, ganoderma lucidum polysaccharide and hydrolyzed casein peptide had synergistic effect on improving sleep.

[0116] From the results of this example, it can be concluded that when ganoderma lucidum polysaccharide, Chinese wingnut powder and hydrolyzed casein peptide are combined at 1~9:1~9:1~9, the sleep improvement has significant synergism, and the sleep improvement rate of the three is better than that of melatonin, and also significantly better than that of the corresponding two combinations, among which Examples 12-15 are close to melatonin in sleep cycle times, sleep duration and average sleep duration.

[0117] Experimental Example 6 (animal verification test)

[0118] The complex of ganoderma lucidum polysaccharide, spina date seed powder and hydrolyzed casein peptide has been proved to have significant sleep improvement effect in zebrafish model. To further determine the effect of the three raw materials on sleep phenotype and biochemical indicators, and to provide more reliable data support for subsequent product development, the complex formula and dose verified by zebrafish experiment were converted into the intake dose of mice according to the dose conversion method reported in patent CN113496071A, and the sleep improvement effect of the complex formula was verified according to the standard method of "Health product function test and evaluation method". The specific results are as follows:

[0119] Table 6 Effect of the complex of ganoderma lucidum polysaccharide, spina date seed powder and hydrolyzed casein peptide on sleep and related indicators of mice

[0120]

[0121] Note: " / " represents no value; statistical difference compared with the model group is marked as * p <0.05, p <0.01, p <0.001.

[0122] As shown in Table 6, diazepam as a positive control, at an appropriate dose, shortened the sleep latency of mice by 38.4%, increased the sleep duration by 145.2%, increased the sleep-promoting neurotransmitter 5-hydroxytryptamine level by 31.5%, increased the ratio of gamma-aminobutyric acid to glutamate by 14.5%, and decreased the excitatory neurotransmitter glutamate by 17.4%. These results indicate that diazepam can significantly improve the sleep phenotype of mice. After the hydrolyzed casein peptide and spina date seed powder (1:2) were compounded with ganoderma lucidum polysaccharide at a ratio of 9:1 and 1:9, the sleep latency was reduced by 17.8% and 22.4%, the sleep duration was increased by 54.6% and 98.4%, the 5-hydroxytryptamine level was increased by 58.2% and 62.0%, the ratio of gamma-aminobutyric acid to glutamate was increased by 4.8% and 8.1%, and the excitatory neurotransmitter glutamate was decreased by 9.5% and 20.6%. The above results indicate that the complex of hydrolyzed casein peptide and spina date seed powder with ganoderma lucidum polysaccharide can improve the sleep of mice.

[0123] After the spina date seed powder and ganoderma lucidum polysaccharide compound powder (1:1) and hydrolyzed casein peptide were compounded at a ratio of 9:1 and 1:9, the sleep latency was reduced by 29.7% and 31.7%, the sleep duration was increased by 114.9% and 122.0%, the 5-hydroxytryptamine level was increased by 67.9% and 71.7%, the ratio of gamma-aminobutyric acid to glutamate was increased by 12.9% and 16.1%, and the excitatory neurotransmitter glutamate was decreased by 24.9% and 29.3%. The above results indicate that the spina date seed powder and ganoderma lucidum polysaccharide compound powder and hydrolyzed casein peptide can improve the sleep of mice.

[0124] 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.

[0125] 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.

[0126] 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".

[0127] Experiment Example 7 (Trial Results for a Population)

[0128] 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.

[0129] 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 total sleep time (A), total sleep time (B), sleep quality (C), and mental state after waking up (D) were all improved. These results show that the Ganoderma lucidum polysaccharide, Suanzaoren powder, and hydrolyzed casein peptide compound beverage can significantly improve mild and moderate sleep disorders, and the sustained improvement effect is good. Figure 2 Figure 2 Figure 2

[0130] Comparative Example

[0131] The raw materials of the comparative example and the results are shown in Table 7. The raw materials were all purchased from the market, and the specific results are as follows:

[0132] Comparative Example 1

[0133] The components in this scheme are (parts by weight): 1 part of licorice powder, 2 parts of jujube powder.

[0134] Comparative Example 2

[0135] The components in this scheme are (parts by weight): 1 part of licorice powder, 3 parts of wheat powder.

[0136] Comparative Example 3

[0137] The components in this scheme are (parts by weight): 2 parts of jujube powder, 3 parts of wheat powder.

[0138] Comparative Example 4

[0139] The components in this scheme are (parts by weight): 1 part of licorice powder, 2 parts of jujube powder, and 3 parts of wheat powder.

[0140] The above comparative examples were subjected to zebrafish experiments by referring to the method of Experimental Example 1, and the experimental results are as follows:

[0141] Table 7 Effect of comparative examples on zebrafish sleep

[0142]

[0143] As can be seen from Table 7, the sleep duration of the licorice powder, wheat powder, and jujube powder single treatment groups increased by 11.5%, 13.0%, and 11.1% respectively compared with the model group, and the sleep improvement rates were 11.8%, 15.4%, and 18.2% respectively, indicating that these raw materials alone have sleep improvement effects. However, whether these raw materials are combined in pairs or in three, the sleep improvement rates increase significantly, but the corresponding CI values are 1.61, 1.52, 1.42, and 1.28 respectively, all greater than 1, proving that the various combinations of these three raw materials do not have a synergistic effect, and also showing that not all combinations of raw materials with sleep improvement effects have a synergistic effect.

[0144] ​​​The above-described embodiments are merely illustrative of several embodiments of the present application, which are described in more detail and in a specific and detailed manner, but should not be construed as limiting the scope of the patent. It should be noted that for those skilled in the art, several modifications and improvements can be made without departing from the concept of the present application, and these all belong to the protection scope of the present application. Therefore, the protection scope of the patent of the present application should be subject to the appended claims.

Claims

1. A composition for improving sleep, characterized by, Components by weight parts: Ganoderma lucidum polysaccharide 0.5-9 parts; hydrolyzed casein peptide 0.3-9 parts; and spina date seed powder 0.5-9 parts; The Ganoderma lucidum polysaccharide has a Ganoderma lucidum polysaccharide mass percentage content greater than 5%; the hydrolyzed casein peptide has a protein hydrolysate with a relative molecular mass less than 1000 Da accounting for not less than 80% and an oligopeptide with a relative molecular mass less than 3000 Da accounting for not less than 90%; and the spina date seed powder has a total spina date glycoside mass percentage content not less than 1%.

2. The composition of claim 1, wherein, The hydrolyzed casein peptide and the spina date seed powder are compounded at a weight ratio of 1:2, and then compounded with the Ganoderma lucidum polysaccharide at a total weight ratio of 9:1-1:

9.

3. The composition of claim 1, wherein The Ganoderma lucidum polysaccharide and the spina date seed powder are compounded at a weight ratio of 1:11, and then compounded with the hydrolyzed casein peptide at a total weight ratio of 9:1-1:

9.

4. The composition of claim 1, wherein The Ganoderma lucidum polysaccharide and the hydrolyzed casein peptide are compounded at a weight ratio of 2:1, and then compounded with the spina date seed powder at a total weight ratio of 9:1-1:

9.

5. A composition for improving sleep, characterized by, The composition is Ganoderma lucidum polysaccharide compounded with spina date seed powder at a weight ratio of 1:9-9:1; the Ganoderma lucidum polysaccharide has a Ganoderma lucidum polysaccharide mass percentage content greater than 5%; and the spina date seed powder has a total spina date glycoside mass percentage content not less than 1%.

6. Use of the composition of any one of claims 1-5 in the preparation of a sleep improvement product.

7. Use according to claim 6, characterized in that, The sleep improvement product is a drug or a health food.

8. Use according to claim 7, characterized in that, The health food is a solid beverage, a pressed tablet candy or an oral liquid.

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