A composition for synergistically improving sleep and use thereof in the preparation of sleep improvement products

Abstract

The application discloses a composition for synergistically improving sleep and application of the composition in preparation of sleep-improving products, components of the composition include green tea extract, ganoderma lucidum polysaccharide and saffron powder, any two components can be compounded in a specific proportion, or the three components can be compounded in a specific proportion, and the sleep of mice is significantly improved, and the improvement effect of the three components is more significant and is equivalent to that of melatonin. It is verified through experiments that the compounded powder of ganoderma lucidum polysaccharide, green tea extract and saffron powder has significant effects on sleep improvement rate, sleep round number and sleep duration by increasing 5-hydroxytryptamine level and gamma-aminobutyric acid / glutamic acid ratio and reducing glutamic acid level, significantly shortens the sleep latency of mice, increases the sleep duration, and thus improves the sleep quality of mice. Important experimental basis and data support are provided for development of a new sleep-improving health-care food.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biological medicine and health food, and particularly relates to a composition for synergistically improving sleep and application thereof in preparation of sleep improvement products. BACKGROUND

[0002] With the acceleration of modern life pace, sleep problems are increasingly prominent. Insufficient sleep not only affects daily life and study, but also can cause various health problems. In recent years, sleep improvement research has become a hot spot. For example, the Chinese patent application with the publication number CN105902536A discloses application of loganin in preparation of sleep improvement drugs or health products. Animal experiments show that loganin can significantly prolong the total sleep time of mice, and has the characteristics of dose-dependent increase in sleep amount. The Chinese patent application with the publication number CN107375374A discloses a fermentation extraction method of American ginseng / ginseng extract with the function of assisting sleep improvement. Through fermentation extraction of American ginseng / ginseng, the large molecular substances such as ginsenosides in American ginseng / ginseng which are not easy to be absorbed are converted into small molecular substances, and the functional food containing GABA is produced through secondary fermentation of American ginseng / ginseng, thereby assisting sleep improvement.

[0003] The sleep improvement products include small molecular drugs, traditional Chinese medicine compositions or medicinal and edible compositions, but chemical drugs are easy to cause side effects, and the compositions have problems such as complex components, complicated preparation process or limited improvement effect. Therefore, it is of great significance to develop a new sleep improvement product which is more efficient and safer. SUMMARY

[0004] The present application provides a composition for synergistically improving sleep and application thereof in preparation of sleep improvement products. The composition has simple components, is convenient to prepare, has a significant improvement effect on sleep disorders, and is safe and reliable.

[0005] Combination index (CI) is a method for quantitatively evaluating the synergistic effect of drugs or active ingredients based on the law of mass action. The method quantifies the synergistic effect through the CI value. When the CI value is less than 1, it indicates that two or more components have a significant synergistic effect, equal to 1 indicates that two or more components have an additive effect, and greater than 1 indicates that two or more components have an antagonistic effect. The present application screens a batch of substances with sleep improvement effect from a series of food raw materials, explores the synergistic effect of a plurality of food raw materials with sleep improvement effect based on the combination index method, and develops a sleep improvement food composition which is more efficient and safer.

[0006] The components of the composition include green tea extract, ganoderma lucidum polysaccharide and saffron powder, which can be any two components compounded in a specific ratio, or three components compounded in a specific ratio, and have a significant improvement effect on the sleep of mice, and the improvement effect of the three components compounded is more significant, and is comparable to the effect of melatonin.

[0007] Saffron powder can regulate central nervous system neurotransmitters such as GABA and 5-hydroxytryptamine, promote sedation and sleep; Ganoderma lucidum polysaccharide affects the secretion of melatonin by the pineal gland, thereby regulating the sleep cycle, and by regulating the nervous system, it can also regulate the levels of neurotransmitters such as GABA and 5-hydroxytryptamine, and improve sleep quality; Theanine rich in green tea extract, as a natural non-protein amino acid, significantly improves sleep quality by regulating neurotransmitter balance, promoting alpha wave activity, and synergistic effect with GABA. Green tea extract and ganoderma lucidum polysaccharide improve sleep quality through multiple pathways such as GABA, 5-hydroxytryptamine and melatonin, and enhance the sleep-promoting effect of the two through saffron powder.

[0008] According to the standard of "Health Food Function Test and Evaluation Method", the sleep improvement effect of the compound is systematically verified, and the compound powder of ganoderma lucidum polysaccharide, green tea extract and saffron powder improves the sleep improvement rate, sleep round number and sleep duration by increasing the level of 5-hydroxytryptamine and the ratio of GABA / glutamate, and reducing the level of glutamate. In a specific ratio, the synergistic effect between two or three is fully played, which significantly shortens the sleep latency of mice, increases the sleep duration, and thus improves the sleep quality of mice. This provides important experimental basis and data support for the development of new sleep-improving health foods.

[0009] The application also evaluates the sleep improvement effect of the pilot production drink based on the composition, and the results show that after continuously taking the compound drink for 7 days, 89% of the subjects significantly shorten the sleep time, 91% of the subjects significantly prolong the total sleep time, 82% of the subjects significantly improve the sleep quality, and 83% of the subjects significantly improve the mental state after waking up. In addition, the sleep data record for 7 consecutive days shows that with the extension of the taking time, the sleep time, total sleep time, sleep quality and mental state after waking up of the subjects show a continuous improvement trend.

[0010] Based on this, the application provides a composition for synergistically improving sleep and its application, which is a compound of any two components of ganoderma lucidum polysaccharide, green tea extract and saffron powder or a compound of three components.

[0011] The sources of each component in the composition can be obtained by purchase or prepared by conventional processing methods in the art. The forms of each component include but are not limited to powder, and can also be extract, extract, concentrate and slice, the extract can be water extract or alcohol extract, preferably water extract, and the skilled person in the art can adjust the compounding according to the actual situation. The preparation method of the composition is conventional mixing, for example, uniformly mixing each powdered raw material.

[0012] When the three components are compounded, the composition of the composition is: Ganoderma lucidum polysaccharide 0.5-10 parts, green tea extract 0.3-10 parts, and saffron powder 0.5-10 parts.

[0013] A preferred scheme for compounding three components, the green tea extract and the saffron powder are compounded at a weight ratio of 2:1, and then compounded with the Ganoderma lucidum polysaccharide at a weight ratio of 1:10-10:1 of the total weight of the two to the weight of the Ganoderma lucidum polysaccharide. Further, the green tea extract and the saffron powder are compounded at a weight ratio of 2:1, and then compounded with the Ganoderma lucidum polysaccharide at a weight ratio of 10:1 of the total weight of the two to the weight of the Ganoderma lucidum polysaccharide.

[0014] Another preferred scheme for compounding three components, the Ganoderma lucidum polysaccharide and the saffron powder are compounded at a weight ratio of 1:1, and then compounded with the green tea extract at a weight ratio of 1:10-10:1 of the total weight of the two to the weight of the green tea extract. Further, the Ganoderma lucidum polysaccharide and the saffron powder are compounded at a weight ratio of 1:1, and then compounded with the green tea extract at a weight ratio of 10:1 of the total weight of the two to the weight of the green tea extract.

[0015] Another preferred scheme for compounding three components, the Ganoderma lucidum polysaccharide and the saffron powder are compounded at a weight ratio of 1:1, and then compounded with the green tea extract at a weight ratio of 1:10-10:1 of the total weight of the two to the weight of the green tea extract. Further, the Ganoderma lucidum polysaccharide and the saffron powder are compounded at a weight ratio of 1:1, and then compounded with the green tea extract at a weight ratio of 10:1 of the total weight of the two to the weight of the green tea extract.

[0016] When the composition is compounded by two components:

[0017] An optional compounding scheme, the green tea extract and the saffron powder are compounded at a weight ratio of 1:8-8:1. The green tea extract and the saffron powder can significantly improve the sleep quality of zebrafish at different compounding ratios, and show significant synergy within the ratio range of 1:8 to 8:1. Further preferably, the weight ratio of the green tea extract to the saffron powder is 1:8, which shows the best synergy.

[0018] In another optional compounding scheme, the Ganoderma lucidum polysaccharide and the saffron powder are compounded at a weight ratio of 1:10 to 10:1. The Ganoderma lucidum polysaccharide and the saffron powder, at a compounding ratio ranging from 1:10 to 10:1, can significantly improve the sleep improvement rate of zebrafish, while increasing the number of sleep rounds, the total sleep time and the average sleep time, and showing a significant synergistic effect. Further preferably, the Ganoderma lucidum polysaccharide and the saffron powder are compounded at a weight ratio of 10:1, showing the best synergistic effect.

[0019] In another optional compounding scheme, the Ganoderma lucidum polysaccharide and the saffron powder are compounded at a weight ratio of 1:10 to 10:1. The Ganoderma lucidum polysaccharide and the saffron powder, at a compounding ratio ranging from 1:10 to 10:1, can significantly improve the sleep improvement rate of zebrafish, while increasing the number of sleep rounds, the total sleep time and the average sleep time, and showing a significant synergistic effect. Further preferably, the Ganoderma lucidum polysaccharide and the saffron powder are compounded at a weight ratio of 10:1, showing the best synergistic effect.

[0020] The application also provides a use of the composition in the preparation of a sleep improvement product.

[0021] Optionally, the sleep improvement product is a pharmaceutical product or a health food.

[0022] Optionally, the sleep improvement includes, but is not limited to, improving the sleep improvement rate, increasing the number of sleep rounds and the sleep time.

[0023] For the preparation of health food, the composition can be prepared with food-acceptable excipients such as solvents, binders, fillers, sugars and sweeteners, etc. according to conventional food preparation methods. The type of health food can be any one of tablets, capsules, oral liquids, solid beverages, pills, granules, decoctions, pastes, lotions, dripping pills or syrups or tea bags.

[0024] Further preferably, when the composition is compounded by Ganoderma lucidum polysaccharide and green tea extract at a ratio of 1:1 and then compounded with saffron powder at a ratio of 1:10, the concentration of the composition in the oral liquid is: Ganoderma lucidum polysaccharide 0.4 mg / mL, green tea extract 0.4 mg / mL, and saffron powder 8 mg / mL.

[0025] For the preparation of pharmaceutical products, the composition can be prepared with pharmaceutically acceptable excipients according to conventional pharmaceutical preparation methods. The pharmaceutically acceptable excipients include buffers, emulsifiers, suspending agents, decomposing agents, disintegrating agents, dispersing agents, binders, excipients, stabilizers, chelating agents, diluents, gelling agents, preservatives, wetting agents, lubricants, absorption retardants, liposomes, etc., which can be selected according to different needs. The dosage forms of the pharmaceutical products include granules, tablets, capsules, granules, granules, oral liquids, dripping pills, micro-pellets or injections, etc.

[0026] In the above health food and medicine, the adding amount and adding ratio of the composition and related adjuvants of the present application can be adjusted according to actual needs.

[0027] Compared with the prior art, the present application has the following beneficial effects:

[0028] (1) The composition of the present application has a significant improvement effect on sleep quality whether combined two by two or three.

[0029] (2) The composition of the present application has simple components, and all are derived from natural food raw materials, which is safe and reliable.

[0030] (3) The preparation process of the composition of the present application is simple and suitable for industrial production. DETAILED DESCRIPTION

[0031] The technical solutions of the present application will be described below in conjunction with specific embodiments. Obviously, the described embodiments are only a part of the embodiments of the present application, not all. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the present application.

[0032] 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 terms used in the specification of the present application are only for the purpose of describing specific embodiments and are not intended to limit the present application.

[0033] Ganoderma lucidum polysaccharide (extract, Ganoderma lucidum polysaccharide content not less than 5%); saffron powder (saffron glycoside content not less than 3.5%); green tea extract (tea amino acid content not less than 20%) used in the following examples are all purchased from the market, all in powder form, and these raw materials all meet the food grade standard, ensuring the safety and effectiveness of the examples. In the composition compounded two by two, the two powder raw materials are mixed according to the ratio; in the composition compounded three, two components are mixed according to a specific ratio and then fully mixed with the third component.

[0034] In addition to the above purchase route, the raw materials of the composition, Ganoderma lucidum polysaccharide, saffron powder and green tea extract, can also be extracted and prepared by themselves:

[0035] The preparation method of saffron powder and Ganoderma lucidum polysaccharide is as follows: the tested green tea, saffron and purple Ganoderma are separately or mixedly pulverized into micro-powder, water or low proportion of ethanol is used as solvent, then according to the appropriate solid-liquid ratio (1:10-1:30), the multi-functional extraction tank is used for cyclic extraction (60℃ or 100℃, 1 hour), gauze filtration, concentration, freeze-drying to obtain powder, which is used as the compounding raw material in the following examples. The above raw materials all meet the food and medicine standards.

[0036] The composition formulations of Examples 1-15 are shown in Table 1:

[0037] The composition formulations of each example in Table 1

[0038]

[0039] Experimental Example 1

[0040] In order to verify the improved sleep effect of the composition of the present application, 6-day-old wild type AB zebrafish were selected for the experiment. A pentylazole-induced insomnia model was used, and melatonin was used as a positive control. The concentration of melatonin was set to 100 μM (see Table 2 for the composition of the raw materials), wherein the green tea extract (EC50 is 45.4 μg·mL -1 ), ganoderma lucidum polysaccharide (extract, EC50 is 164.6 μg·mL -1 ) and saffron powder (EC50 is 322.4 μg·mL -1 ) were set at their respective EC50 values, the dose of vitamin B6 was set to 0.2 μg·mL -1 , the dose of rosemary extract was set to 300 μg·mL -1 , and the dose of hops extract was set to 400 μg·mL -1 .

[0041] Before the start of the behavior experiment, pentylazole solution was added to the water in each group, and melatonin stock solution and treatment group stock solution were added in a timely manner, so that the dose met the experimental dose setting.

[0042] The behavior analysis instrument was used to record the movement trajectory of zebrafish within 1 hour, and the movement speed was classified as follows: less than 0.5 cm / s was slow, 0.5-2 cm / s was medium, and more than 2 cm / s was high. The duration of slow movement within 1 hour ≥6 seconds was recorded as one sleep. The total number of sleep rounds within 1 hour was determined by counting the total number of sleep. All sleep times were added to obtain the total sleep time, and the average sleep time (i.e., the average value of the total sleep time) was calculated.

[0043] In order to quantitatively evaluate the sleep improvement effect, the following formula was used to calculate the sleep improvement rate:

[0044] Sleep improvement rate = 100 x (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.

[0045] The results are shown in Table 2:

[0046] Table 2: Raw materials for improving sleep composite formulation based on zebrafish insomnia model

[0047]

[0048] Note: " / " means no value; statistical differences compared with the model group are marked as p <0.05, p <0.01, p <0.001; statistical differences compared with the control group are marked as p <0.05, p <0.01, p <0.001.

[0049] The present test example aims to evaluate the improvement effect of various food raw materials on insomnia phenotype and explore their synergistic effect to develop a sleep improvement composite formula with synergistic effect. In this test example, 6 kinds of food raw materials are selected, including ganoderma lucidum polysaccharide, green tea extract, saffron powder, vitamin B6, rosemary extract and hops extract, which are also purchased from the market. The sleep improvement effect of the above-mentioned raw materials is systematically studied. The research results are as follows:

[0050] Sleep improvement rate: Ganoderma lucidum polysaccharide > saffron powder > green tea extract > hops extract > vitamin B6 > rosemary extract;

[0051] Sleep round number: green tea extract > ganoderma lucidum polysaccharide > saffron powder > hops extract > vitamin B6 > rosemary extract;

[0052] Sleep duration: saffron powder > green tea extract > ganoderma lucidum polysaccharide > hops extract > vitamin B6 > rosemary extract.

[0053] The research results show that ganoderma lucidum polysaccharide and saffron powder have significant effect on improving sleep improvement rate, and the two may have synergistic effect, which can reduce the effective dosage. Ganoderma lucidum polysaccharide and green tea extract have significant effect on increasing sleep round number, and the two may have synergistic effect after compounding. Green tea extract and saffron powder have significant effect on prolonging sleep duration, and the two may have synergistic effect after compounding.

[0054] Considering the improvement effect and contribution rate of each raw material, the present application selects ganoderma lucidum polysaccharide, saffron powder and green tea extract as the core raw materials of the sleep improvement composite formula. On the basis of verifying the synergistic effect of the two raw materials, the third raw material is further compounded to verify the synergistic effect of the three raw materials.

[0055] The goal of the present application is to develop a sleep improvement composite formula with synergistic effect. The subsequent examples will focus on this goal, and through experimental verification of the synergistic effect of different raw material combinations, the formula proportion will be optimized to achieve more efficient and safer sleep improvement effect. The composite formula of the present application is expected to provide a new, natural food raw material-based solution for improving sleep disorders.

[0056] Experimental Example 2

[0057] The experimental method was the same as that in Experimental Example 1, and the compounding raw materials and results are shown in Table 3:

[0058] Table 3 Effects of green tea extract and saffron powder compound on sleep of zebrafish

[0059]

[0060] Note: The dosage of Example 1, Example 2 and Example 3 was 50% effective dose; " / " represents no value; statistical difference compared with the model group is marked as * p <0.05,** p <0.01,*** p <0.001.

[0061] As shown in Table 3, the sleep duration of zebrafish treated with green tea extract and saffron powder alone was increased by 59.3% and 63.0% respectively compared with the model group. Based on the calculation results of half effective concentration (EC50), the compound ratio of green tea extract and saffron powder was set as 2:1, 1:8 and 8:1. The experimental results showed that the improvement effect of sleep duration of zebrafish by different proportions of compound was 67.7% (2:1), 81.9% (1:8) and 85.9% (8:1) respectively. The corresponding combination index (CI value) was 0.68, 0.59 and 0.62 respectively, all less than 1, indicating that within the ratio range of 1:8 to 8:1, green tea extract and saffron powder had synergistic improvement effect on sleep duration. In addition, the improvement effect of sleep round number of zebrafish by different proportions of compound was 34.9% (2:1), 40.3% (1:8) and 45.0% (8:1) respectively. The corresponding sleep improvement rate was 39.2%, 46.3% and 52.4% respectively, and the average sleep duration was increased by 24.3%, 29.4% and 28.2% respectively. These results further confirmed the significant improvement effect of green tea extract and saffron powder compound on sleep of zebrafish.

[0062] In summary, green tea extract and saffron powder could significantly improve the sleep quality of zebrafish at different compounding ratios, and showed significant synergistic effect within the ratio range of 1:8 to 8:1, but there was still a certain gap with the positive control melatonin. This finding provides an important experimental basis for developing sleep improvement compound formula based on green tea extract and saffron powder.

[0063] Experimental Example 3

[0064] The experimental method was the same as that in Experimental Example 1, and the compounding raw materials and results are shown in Table 4:

[0065] Table 4 Effects of Ganoderma lucidum polysaccharide and saffron powder compound on sleep of zebrafish

[0066]

[0067] Note: The dosage of Example 4, Example 5 and Example 6 is 50% effective dose; " / " means no value; statistical difference compared with the model group is marked as * p <0.05,** p <0.01,*** p <0.001.

[0068] According to the data in Table 4, the sleep improvement rates of zebrafish treated with Ganoderma lucidum polysaccharide and saffron powder alone were 43.5% and 37.8%, respectively. Based on the half effective concentration (EC50) value, three compound ratios were set: 1:1, 10:1 and 1:10. The experimental results showed that the sleep improvement rates of zebrafish treated with different ratios of Ganoderma lucidum polysaccharide and saffron powder compound were 45.6% (1:1), 50.4% (10:1) and 57.3% (1:10), respectively. The corresponding combination index (CI value) were 0.49, 0.53 and 0.62, respectively, all less than 1, indicating that within the ratio range of 1:10 to 10:1, the Ganoderma lucidum polysaccharide and saffron powder compound had a significant synergistic effect on the sleep improvement rate. In addition, the sleep round times of zebrafish treated with different ratios of compound were increased by 36.6% (1:1), 39.2% (10:1) and 37.3% (1:10), respectively, the sleep duration was increased by 70.2% (1:1), 83.8% (10:1) and 86.6% (1:10), respectively, and the average sleep duration was increased by 24.7% (1:1), 32.2% (10:1) and 36.2% (1:10), respectively. These results further confirmed the significant improvement of Ganoderma lucidum polysaccharide and saffron powder compound on the sleep quality of zebrafish.

[0069] In summary, within the ratio range of 1:10 to 10:1, Ganoderma lucidum polysaccharide and saffron powder can significantly improve the sleep improvement rate of zebrafish, while increasing the sleep round times, total sleep duration and average sleep duration, and showing a significant synergistic effect, in which the sleep round times and average sleep duration are close to the positive control melatonin. This finding provides important experimental basis for the development of sleep improvement compound formula based on Ganoderma lucidum polysaccharide and saffron powder, and has important application value.

[0070] Experimental Example 4

[0071] The experimental method is the same as that in Experimental Example 1, and the compound raw materials and results are shown in Table 5:

[0072] Table 5 Effects of Ganoderma lucidum polysaccharide and green tea extract compound on sleep of zebrafish

[0073]

[0074] Note: The dosage of Example 7, Example 8 and Example 9 is 50% effective dose; " / " means no value; statistical difference compared with the model group is marked as * p <0.05,** p <0.01,*** p <0.001.

[0075] According to the data in Table 5, the sleep round times of zebrafish treated with Ganoderma lucidum polysaccharide and green tea extract alone were increased by 27.1% and 24.5% respectively compared with the model group, showing a significant potential to improve sleep round times. Based on the half effective concentration (EC50) value, three compound ratios were set: 1:1, 9:1 and 1:9. The experimental results showed that the improvement effect of different ratios of Ganoderma lucidum polysaccharide and green tea extract complex on the sleep round times of zebrafish was 30.3% (1:1), 34.2% (9:1) and 39.4% (1:9) respectively. The corresponding joint index (CI value) was 0.72, 0.69 and 0.59 respectively, all less than 1, indicating that within the ratio range of 1:9 to 9:1, the Ganoderma lucidum polysaccharide and green tea extract complex had a synergistic improvement effect on sleep round times. In addition, the sleep improvement rates of zebrafish treated with different ratios of complex were 41.5% (1:1), 48.3% (9:1) and 55.4% (1:9) respectively, the sleep duration was increased by 1.7 times (1:1), 1.8 times (9:1) and 1.8 times (1:9) respectively, and the average sleep duration was increased by 33.5% (1:1), 36.4% (9:1) and 31.8% (1:9) respectively. These results further confirmed the significant improvement effect of Ganoderma lucidum polysaccharide and green tea extract complex on the sleep quality of zebrafish. In summary, within the compound ratio range of 1:9 to 9:1, Ganoderma lucidum polysaccharide and green tea extract can significantly improve the sleep round times, sleep improvement rate, sleep duration and average sleep duration of zebrafish, and show a synergistic effect, which is close to the positive control melatonin in improving sleep round times and average sleep duration. This finding provides important experimental basis for the development of sleep improvement complex formula based on Ganoderma lucidum polysaccharide and green tea extract, and has important application value and market potential.

[0076] Experimental Example 5

[0077] The experimental method is the same as that in Experimental Example 1, and the compound raw materials and results are shown in Table 6:

[0078] Table 6 Effect of Ganoderma lucidum polysaccharide, green tea extract and saffron powder complex on sleep of zebrafish

[0079]

[0080] 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.

[0081] According to the data in Table 6, the sleep improvement rates of zebrafish treated with Ganoderma lucidum polysaccharide, green tea extract, and saffron pollen individually were 43.7%, 27.5%, and 33.7%, respectively. These treatments significantly increased the number of sleep rounds, sleep duration, and average sleep duration compared to the model group, demonstrating significant potential for improving sleep.

[0082] Based on the above experimental results, this study further explored the synergistic effects of pairwise and triadic combinations of Ganoderma lucidum polysaccharides, green tea extract, and saffron pollen. The experiments used fixed-ratio compound powders as follows: green tea extract and saffron pollen compound powder (2:1); Ganoderma lucidum polysaccharides and saffron pollen compound powder (1:1); and Ganoderma lucidum polysaccharides and green tea extract compound powder (1:1). The results showed that the sleep improvement rates of the above compound powders were 42.6%, 46.8%, and 50.6%, respectively, with corresponding combination indices (CI values) of 0.65, 0.59, and 0.57, all less than 1, indicating that pairwise combinations have a significant synergistic effect. In addition, the effects of the two-component powders on the number of sleep rounds, sleep duration, and average sleep duration were as follows: the number of sleep rounds increased by 24.8%, 28.0%, and 29.3%; sleep duration increased by 63.4%, 64.9%, and 68.9%; and average sleep duration increased by 31.0%, 28.7%, and 30.4%.

[0083] Further experiments showed that when the compound powder of green tea extract and saffron pollen (2:1) was combined with Ganoderma lucidum polysaccharide at a ratio of 10:1 (Example 10): the number of sleep rounds increased by 31.2%, sleep duration increased by 74.6%, average sleep duration increased by 32.7%, sleep improvement rate was 65.8%, and CI value was 0.52; when combined at a ratio of 1:10 (Example 11): the number of sleep rounds increased by 33.1%, sleep duration increased by 75.6%, average sleep duration increased by 31.6%, sleep improvement rate was 68.3%, and CI value was 0.50.

[0084] When the Ganoderma lucidum polysaccharide and saffron powder (1:1) were compounded with the green tea extract, the proportion of 10:1 (Example 12) was compounded: the sleep round number increased by 35.7%, the sleep duration increased by 81.2%, the average sleep duration increased by 33.3%, the sleep improvement rate was 69.8%, and the CI value was 0.43; the proportion of 1:10 (Example 13) was compounded: the sleep round number increased by 36.9%, the sleep duration increased by 85.4%, the average sleep duration increased by 35.1%, the sleep improvement rate was 71.6%, and the CI value was 0.50.

[0085] When the Ganoderma lucidum polysaccharide and green tea extract powder (1:1) were compounded with the saffron powder, the proportion of 10:1 (Example 14) was compounded: the sleep improvement rate was 72.3%, the CI value was 0.35; the sleep round number increased by 39.5%, the sleep duration increased by 92.0%, and the average sleep duration increased by 37.4%; the proportion of 1:10 (Example 15) was compounded: the sleep improvement rate was 75.2%, the CI value was 0.32; the sleep round number increased by 42.0%, the sleep duration increased by 97.1%, and the average sleep duration increased by 38.6%. The CI values of all compound combinations were less than 1, indicating that the three compounds had significant synergy, and the improvement effect was comparable to that of the positive control melatonin.

[0086] In summary, the Ganoderma lucidum polysaccharide, green tea extract, and saffron powder can significantly improve the sleep quality of zebrafish when compounded in the ratio of 1:10 to 10:1, including significantly increasing the sleep improvement rate, increasing the sleep round number, prolonging the sleep duration, and increasing the average sleep duration, and showing significant synergy. This finding provides important experimental evidence for the development of sleep improvement composite formulations based on Ganoderma lucidum polysaccharide, green tea extract, and saffron powder, and has important application value and market potential.

[0087] Experimental Example 6

[0088] To clarify the effects of Ganoderma lucidum polysaccharide, green tea extract, and saffron powder complex on sleep phenotype and biochemical indicators, and to provide scientific basis for subsequent product development, based on the compound formula and dose verified in the previous zebrafish experiment, the dose was converted to be suitable for mice according to the dose conversion method described in the patent CN113496071A. On this basis, the sleep improvement effect of the complex was systematically verified according to the standard of "Health Food Function Test and Evaluation Method". The specific experimental results are as follows:

[0089] Table 7 Improvement effect of Ganoderma lucidum polysaccharide, green tea extract, and saffron powder complex on sleep and related indicators of mice

[0090]

[0091] Note: " / " indicates no numerical value; statistical differences compared to the model group are marked with *. p <0.05,** p <0.01, *** p <0.001.

[0092] According to the data in Table 7, diazepam, as a positive control, shortened the sleep latency by 32.0%, increased sleep duration by 97.0%, increased serotonin levels by 30.4%, increased the γ-aminobutyric acid / glutamate ratio by 15.4%, and decreased glutamate levels by 22.7% in mice. These results indicate that diazepam can significantly improve the sleep phenotype in mice.

[0093] In this experiment, we further explored the effects of a compound powder of Ganoderma lucidum polysaccharides, green tea extract, and saffron pollen on sleep in mice. The experimental results are as follows:

[0094] A compound powder of green tea extract and saffron pollen (2:1) was combined with Ganoderma lucidum polysaccharide at a ratio of 10:1 (Example 10): Sleep latency decreased by 9.9%, sleep duration increased by 34.3%, serotonin levels increased by 3.8%, the γ-aminobutyric acid / glutamate ratio increased by 3.1%, and glutamate levels decreased by 6.2%. A compound powder of 1:10 (Example 11): Sleep latency decreased by 11.6%, sleep duration increased by 52.7%, serotonin levels increased by 7.8%, the γ-aminobutyric acid / glutamate ratio increased by 6.2%, and glutamate levels decreased by 11.6%.

[0095] A compound powder of Ganoderma lucidum polysaccharides and saffron pollen (1:1) was compounded with green tea extract at a ratio of 10:1 (Example 12): Sleep latency decreased by 15.0%, sleep duration increased by 64.2%, serotonin levels increased by 12.8%, the γ-aminobutyric acid / glutamate ratio increased by 4.6%, and glutamate levels decreased by 13.5%. A compound powder at a ratio of 1:10 (Example 13): Sleep latency decreased by 18.5%, sleep duration increased by 76.6%, serotonin levels increased by 17.1%, the γ-aminobutyric acid / glutamate ratio increased by 9.2%, and glutamate levels decreased by 15.1%.

[0096] A compound powder of Ganoderma lucidum polysaccharides and green tea extract (1:1) was compounded with saffron powder at a ratio of 10:1 (Example 14): Sleep latency decreased by 26.2%, sleep duration increased by 82.7%, serotonin levels increased by 21.9%, the γ-aminobutyric acid / glutamate ratio increased by 10.8%, and glutamate levels decreased by 21.3%. A compound powder at a ratio of 1:10 (Example 15): Sleep latency decreased by 30.3%, sleep duration increased by 93.7%, serotonin levels increased by 28.3%, the γ-aminobutyric acid / glutamate ratio increased by 13.8%, and glutamate levels decreased by 20.2%.

[0097] In summary, the Ganoderma lucidum polysaccharide, green tea extract and saffron powder compound powder significantly shortens the sleep latency of mice, increases the sleep duration, and improves the sleep quality of mice by increasing the 5-hydroxytryptamine level and the ratio of gamma-aminobutyric acid / glutamate and reducing the glutamate level. According to the standard of "Health Food Function Test and Evaluation Method", the composition of the application has a significant sleep improvement effect, which provides important experimental basis and data support for the development of new sleep improvement health food.

[0098] Experimental Example 6

[0099] In order to evaluate the sleep improvement effect of the pilot production drink (the formula is Example 15, wherein 10 mg of Ganoderma lucidum polysaccharide, 10 mg of green tea extract and 200 mg of saffron powder are dissolved in 25 ml of pure water, and the sterilized package is provided to volunteers, who take it 10-30 minutes before sleep every day) in the application, 60 volunteers were recruited by unit work group, and the Pittsburgh Sleep Quality Index (PSQI) questionnaire and polysomnography (PSG) were used to evaluate the sleep quality. After strict screening, 27 volunteers with PSQI scores of 7-10 (mild sleep disorder) and 10-15 (moderate sleep disorder) were finally selected as test objects. Since this experiment mainly focuses on the intervention effect of the pilot production drink, no control group was set, but the sleep improvement effect of the test population after continuously taking the drink for 7 days was tracked. After the experiment, 27 valid questionnaires were collected. The statistical results are as follows:

[0100] Table 8 Effect of Example 15 formula on sleep improvement of test population and 7-day continuous sleep improvement effect

[0101]

[0102] According to the data in Table 8, after the test subjects continuously took the Ganoderma lucidum polysaccharide, green tea extract and saffron powder compound drink for 7 days, the results showed that 89% of the test subjects had significantly shortened sleep latency, 91% of the test subjects had significantly prolonged total sleep time, 82% of the test subjects had significantly improved sleep quality, and 83% of the test subjects had significantly improved mental state after waking up. In addition, the 7-day continuous sleep data records showed that as the length of time of taking the drink increased, the sleep latency, total sleep time, sleep quality and mental state after waking up of the test subjects all showed a continuous improvement trend.

[0103] In summary, the compound drink of the application has a significant improvement effect on mild and moderate sleep disorders, and as the length of time of taking the drink increases, the improvement effect continuously improves. This result further confirms the effectiveness of the Ganoderma lucidum polysaccharide, green tea extract and saffron powder compound formula in improving sleep, which provides important clinical basis and data support for the development of new sleep improvement health food.

[0104] Comparative Example

[0105] The raw materials and results of the comparative examples are shown in Table 8. The raw materials were all purchased from the market. The specific experimental design is as follows:

[0106] Comparative Example 1

[0107] The components (parts by weight) in this comparative example are as follows: sour cherry extract 2 parts, tomato powder 5 parts.

[0108] Comparative Example 2

[0109] The components (parts by weight) in this comparative example are as follows: sour cherry extract 1 part, tomato powder 10 parts.

[0110] Comparative Example 3

[0111] The components (parts by weight) in this comparative example are as follows: sour cherry extract 10 parts, tomato powder 1 part.

[0112] Comparative Example 4

[0113] The components (parts by weight) in this comparative example are as follows: longan powder 1 part, sour cherry extract 1 part, and tomato powder 10 parts.

[0114] The above comparative examples were subjected to zebrafish experiments according to the method of Experimental Example 1. The experimental results are as follows:

[0115] Table 9. Effect of comparative examples on zebrafish sleep

[0116]

[0117] According to the data in Table 9, when longan powder, sour cherry extract, and tomato powder were treated alone, the sleep duration was increased by 3.7%, 11.8%, and 18.3% compared to the model group, respectively, and the corresponding sleep improvement rates were 2.4%, 14.5%, and 17.6%, respectively. The above results show that these raw materials have certain sleep improvement effects when used alone. However, when combined in pairs or three, although the sleep improvement rate increased, the combination index (CI value) was 1.92, 1.85, 1.79, and 1.65, respectively, all greater than 1. This indicates that these combinations do not exhibit a synergistic effect, and also shows that a single raw material with sleep improvement effect does not necessarily produce a synergistic effect when combined.

[0118] The above examples only express several embodiments of the present application, and the description is more specific and detailed, but it should not be understood as limiting the scope of the patent. It should be noted that for ordinary skilled persons in the art, without departing from the concept of the present application, several modifications and improvements can be made, which are within the scope of the present application. Therefore, the protection scope of the present patent should be subject to the appended claims.

Claims

1. A composition for synergistically improving sleep, characterized in that, It is a compound composed of Ganoderma lucidum polysaccharides, green tea extract, and saffron powder; the Ganoderma lucidum polysaccharides contain no less than 5% by mass; the saffron powder contains no less than 3.5% by mass of crocin; and the green tea extract contains no less than 20% by mass of theanine. The green tea extract and saffron powder are compounded at a weight ratio of 2:1, and then compounded with Ganoderma lucidum polysaccharide at a weight ratio of 1:10 to 10:

1. or The Ganoderma lucidum polysaccharide and saffron powder are compounded in a weight ratio of 1:1, and then compounded with green tea extract in a weight ratio of 1:10 to 10:

1. or Ganoderma lucidum polysaccharide and green tea extract were compounded in a weight ratio of 1:1, and then compounded with saffron pollen in a weight ratio of 1:10 to 10:

1.

2. The composition according to claim 1, characterized in that, The green tea extract and saffron powder were compounded at a weight ratio of 2:1, and then compounded with Ganoderma lucidum polysaccharide at a weight ratio of 10:

1.

3. The composition according to claim 1, characterized in that, The Ganoderma lucidum polysaccharide and saffron powder were compounded in a weight ratio of 1:1, and then compounded with green tea extract in a weight ratio of 10:

1.

4. The composition according to claim 1, characterized in that, Ganoderma lucidum polysaccharide and green tea extract were compounded in a weight ratio of 1:1, and then compounded with saffron pollen in a weight ratio of 1:10 or 10:

1.

5. The use of the composition according to any one of claims 1 to 4 in the preparation of sleep-improving products.

6. The application according to claim 5, characterized in that, The sleep product mentioned is a medicine or health food.

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