Composition for improving sleep quality and use thereof

Abstract

A composition for improving sleep quality includes lactic acid bacteria strains and a carrier. The lactic acid bacteria strains include isolated Bifidobacterium breveBBr-28, Bifidobacterium animalissubsp. lactisPS23, Bifidobacterium longumsubsp. infantisBR631, Bifidobacterium bifidumBD-188 and Lactiplantibacillus plantarumPQQ-1, all of which are deposited at the Food Industry Research and Development Institute. A use of a lactic acid bacteria strain in preparing a composition for improving sleep quality is also disclosed.

Description

[Technical Field]

[0001] This invention relates to a composition and its use, and more particularly to a composition and its use for improving sleep quality. [Previous Technology]

[0002] Sleep quality not only affects physical health but also leads to premature aging; therefore, it is a crucial factor in maintaining normal daily functioning. However, with the prevalence of smartphones and other electronic devices, many people continue to use their phones before bed, such as browsing social media, watching dramas, or playing games. These behaviors significantly reduce sleep quality and contribute to a year-on-year increase in insomnia. Therefore, insomnia is no longer limited to the elderly; it is increasingly affecting younger people, peaking in those aged 20 to 40.

[0003] In view of this, there is an urgent need to develop a safe and long-term edible nutritional supplement that can improve sleep quality and alleviate sleep problems. Since lactic acid bacteria are generally considered safe, identifying compositions containing lactic acid bacteria strains with sleep-enhancing activity is a pressing goal. [Summary of the Invention]

[0004] The present invention provides a composition containing a strain of lactic acid bacteria, which has the physiological activity effect of improving sleep quality. Therefore, the composition of the present invention can be used to improve sleep quality and exists in the form of a food composition or a pharmaceutical composition.

[0005] An embodiment of the present invention provides a composition for improving sleep quality, comprising a strain of lactic acid bacteria having an active effect of improving sleep quality and a carrier. The lactic acid bacteria strains with active effects in improving sleep quality include the isolated *Bifidobacterium breve* strain BBr-28 (accession number BCRC 911254); *Bifidobacterium animalis subsp. lactis* strain PS23 (accession number BCRC 911251); *Bifidobacterium longum subsp. infantis* strain BR631 (accession number BCRC 911172); *Bifidobacterium bifidum* strain BD-188 (accession number BCRC 911252); and *Lactiplantibacillus plantarum* strain PQQ-1 (accession number BCRC 911262). All of these strains are registered with the Food Industry Development and Research Institute.

[0006] Another embodiment of the present invention is the use of a lactic acid bacteria strain in the preparation of a composition for improving sleep quality, wherein the composition comprises a lactic acid bacteria strain having an activity effect of improving sleep quality and a carrier. The lactic acid bacteria strains with active effects in improving sleep quality include the isolated *Bifidobacterium breve* strain BBr-28 (accession number BCRC 911254); *Bifidobacterium animalis subsp. lactis* strain PS23 (accession number BCRC 911251); *Bifidobacterium longum subsp. infantis* strain BR631 (accession number BCRC 911172); *Bifidobacterium bifidum* strain BD-188 (accession number BCRC 911252); and *Lactiplantibacillus plantarum* strain PQQ-1 (accession number BCRC 911262). All of these strains are registered with the Food Industry Development and Research Institute.

[0007] The following detailed description, with reference to specific embodiments and accompanying drawings, will make it easier to understand the purpose, technical content, features and effects of the present invention.

Implementation Method

[0008] The various embodiments of the present invention will be described in detail below, with illustrations provided. In addition to these detailed descriptions, the present invention can be widely implemented in other embodiments. Any easy substitutions, modifications, or equivalent changes to the described embodiments are included within the scope of the present invention and are subject to the claims. In the description of the specification, many specific details are provided to give the reader a more complete understanding of the present invention; however, the present invention may still be implemented even if some or all of the specific details are omitted. Furthermore, well-known steps or elements are not described in the details to avoid unnecessarily limiting the present invention. Identical or similar elements in the drawings will be represented by the same or similar symbols. It should be noted that the drawings are for illustrative purposes only and do not represent the actual size or number of elements; some details may not be fully drawn for the sake of simplicity.

[0009] There is extensive research on the effects of probiotics on human health. Generally speaking, strains that have special effects on human health are called functional probiotics (Guidelines for the evaluation of probiotics in food; Report of joint FAO / WHO working group on drafting guidelines for the evaluation of probiotics in food; London Ontario, Canada April 30 and May 1, 2002: 1-7). It should be noted that the special effects of the above-mentioned functional probiotics lie in the specificity of the strain rather than the species. That is, even the same species may have different effects, or even opposite effects, if they are different strains.

[0010] The freeze-dried culture of the lactic acid bacteria strain described in this invention has been deposited with the Food Industry Development and Research Institute, located at No. 331, Food Rd., Hsinchu City, Taiwan. Detailed information regarding the deposit is shown in Table 1.

[0011] Table 1. Preservation data of lactic acid bacteria strains strain name Classification Deposit number Deposit Date BBr-28 Bifidobacterium breve Bifidobacterium breve BCRC 911254 2024 / 12 / 10 PS23 Bifidobacterium animalis subsp. lactis Bifidobacterium animalissubsp.lactis BCRC 911251 2024 / 12 / 10 BR631 Bifidobacterium longum infant subspecies Bifidobacterium longumsubsp.infantis BCRC 911172 2023 / 02 / 20 BD-188 Bifidobacterium bifidum Bifidobacterium bifidum BCRC 911252 2024 / 12 / 10 PQQ-1 Lactobacillus plantarum Lactiplantibacillus plantarum BCRC 911262 2025 / 01 / 09

[0012] The combination of lactic acid bacteria strains listed in Table 1 has physiological activity that improves sleep quality, so the combination of the above-mentioned lactic acid bacteria strains can be used to improve sleep quality.

[0013] One embodiment of the present invention provides a composition for improving sleep quality, comprising a strain of lactic acid bacteria having the activity of improving sleep quality and a carrier. In one embodiment, the lactic acid bacteria strains include the following isolated strains: Bifidobacterium breve BBr-28 (accession number BCRC 911254); Bifidobacterium animalis subsp. lactis PS23 (accession number BCRC 911251); Bifidobacterium longum subsp. infantis BR631 (accession number BCRC 911172); Bifidobacterium bifidum BD-188 (accession number BCRC 911252); and Lactiplantibacillus plantarum PQQ-1 (accession number BCRC 911262). All of these strains are deposited at the Food Industry Development Institute. In one embodiment, the carrier may be a physiologically acceptable excipient or diluent. Alternatively, the carrier may be a pharmaceutically acceptable excipient or diluent.

[0014] In one embodiment, the effective viable count of the lactic acid bacteria strain in the composition of the present invention is greater than or equal to 1 × 10⁶ CFU / g; in a preferred embodiment, the effective viable count of the lactic acid bacteria strain in the composition of the present invention is equal to or greater than 5 × 10⁹ CFU / g. In one embodiment, the dosage form of the composition of the present invention may be tablets, capsules, or powder for convenient oral administration. Characteristics of the lactic acid bacteria strain of the present invention:

[0015] The taxonomic characteristics of the strain were confirmed based on 16S rRNA sequence analysis and API bacterial identification system analysis results. The morphological and general characteristics of the lactic acid bacteria strain of this invention are detailed in Table 2: Table 2 Morphological and General Characteristics of the Lactic Acid Bacteria Strain of this Invention strain name Morphological characteristics Bifidobacterium breve BBr-28 strain 1. When cultured in MRS medium, the bacteria exhibit irregular shapes such as Y-shape, V-shape, or curved shape. The bacteria are small and usually appear alone, not in chains. 2. Gram-positive bacilli that do not produce spores, do not possess catalase, oxidase, or motility, grow in an absolutely anaerobic environment, with an optimal growth temperature of 37±1℃, and are facultative heterogeneous fermenting strains that do not produce gas during glucose metabolism. Bifidobacterium animalis subsp. lactis PS23 strain 1. When cultured in MRS medium, the bacteria exhibit irregular shapes such as Y-shape, V-shape, or curved shape, and usually appear alone rather than in chains. 2. Gram-positive bacilli that do not produce spores, do not possess catalase, oxidase, or motility, grow in an absolutely anaerobic environment, with an optimal growth temperature of 37±1℃, and are facultative heterogeneous fermenting strains that do not produce gas during glucose metabolism. Bifidobacterium longum infant subspecies BR631 strain 1. When cultured in MRS medium, the bacterial cells are short rod-shaped with an oval tail, usually appearing alone or in pairs of short chains. 2. Gram-positive bacilli that do not produce spores, do not possess catalase, oxidase, or motility, can grow in both aerobic and anaerobic environments, with an optimal growth temperature of 37±1℃. They are facultative heterogeneous fermenting strains that do not produce gas during glucose metabolism. Bifidobacterium bifidum BD-188 1. When cultured in MRS medium, the bacterial cell morphology is medium to short rod-shaped with an oval tail. They usually appear alone or in pairs of short chains. 2. Gram-positive bacilli that do not produce spores, do not possess catalase, oxidase, or motility, can grow in both aerobic and anaerobic environments, with an optimal growth temperature of 37±1℃. They are facultative heterogeneous fermenting strains that do not produce gas during glucose metabolism. Lactobacillus plantarum PQQ-1 1. When cultured in MRS medium, the colonies that grow are white, solid, and round, with short rod-shaped cells and rounded tails, and usually appear alone. 2. Gram-positive bacilli that do not produce spores, do not possess catalase, oxidase, or motility, can grow in both aerobic and anaerobic environments, with an optimal growth temperature of 37±1℃. They are facultative heterogeneous fermenting strains that do not produce gas during glucose metabolism. Preparation of the lactic acid bacteria powder and composition of the present invention:

[0016] The *Bifidobacterium breve* BBr-28 strain, *Bifidobacterium animalis* subsp. *lactobacter* PS23 strain, *Bifidobacterium longum* subsp. *infant* BR631 strain, and *Bifidobacterium bifidum* BD-188 strain of the present invention were isolated from breast milk; the *Lactobacillus plantarum* PQQ-1 strain was isolated from *Saussurea involucrata*. The isolated lactic acid bacteria strains were stored at -80°C with 20% glycerol. In use, 1% (v / v) inoculum was inoculated into Difco TMMRS (De Man, Rogosa and Sharpe) medium and cultured at 37°C for 20 hours. Afterwards, the five cultures were centrifuged at 4°C for 20 minutes, the supernatant was discarded, and the resulting precipitates were freeze-dried to obtain the bacterial powder of each lactic acid bacteria strain. In one embodiment, the precipitate after centrifugation could be mixed with a protective carrier before freeze-drying to maintain better bacterial activity.

[0017] The above-mentioned lactic acid bacteria powder and carrier of the present invention are mixed evenly to form the composition of the present invention, wherein the effective viable count of the lactic acid bacteria strain is equal to or greater than 5 × 10⁹ CFU / g. In one embodiment, the carrier may contain oligosaccharides, sugar alcohols, sodium carboxymethyl cellulose, ethyl cellulose, galactooligosaccharides, stearic acid, magnesium stearate, potato starch, or a combination thereof. For example, oligosaccharides may contain fructooligosaccharides, isomaltose oligosaccharides, xylooligosaccharides, stachyose, mannose, arabinose, or a combination thereof; sugar alcohols may contain erythritol, sorbitol, isomaltitol, xylitol, mannitol, or a combination thereof. Physiological activity analysis of the composition of the present invention:

[0018] First, the Pittsburgh Sleep Quality Index (PSQI) questionnaire was used to assess the sleep of the subjects. The Pittsburgh Sleep Quality Index (PSQI) was proposed by American scholars Buysse et al., and has seven aspects of scores. The higher the PSQI score, the worse the sleep quality. When PQSI > 5, it is assessed as having a sleep disorder. The 30 subjects participating in the experiment were divided into two groups. One group took the capsules of the composition of the present invention twice a day, once between 2-3 pm and once 2 hours before bedtime, for 2 weeks. The other group took the capsules of the composition containing only the powder of Bifidobacterium animalis subsp. lactis PS23, as a control group, and the other methods of administration were the same. Before taking the composition of the embodiment of the present invention and the control group, and after taking it for 2 weeks, the subjects' blood was collected to detect changes in serotonin, and the improvement of the subjects' sleep quality was observed using a wearable medical watch sleep analyzer and the PSQI questionnaire. (I) Effect of taking the composition of an embodiment of the present invention on serotonin:

[0019] Serotonin, also known as 5-hydroxytryptamine (5-HT), is a monoamine neurotransmitter whose functions include regulating mood, appetite, and sleep. Studies have shown that serotonin deficiency can easily lead to lethargy, mood swings, anxiety, insufficient melatonin, or insomnia. Therefore, increasing serotonin levels can help improve sleep quality.

[0020] Before and 2 weeks after taking the compositions of the embodiments and control groups of the present invention, approximately 5 ml of blood was collected from the subjects for serotonin detection. The collected blood was centrifuged to separate the serum. The serum was processed using serotonin ELISA kits (abnova) according to the manufacturer's instructions, and after HRP (horseradish peroxidase) catalyzed TMB (3,3',5,5'-Tetramethyl-benzidine) color development, the absorbance was measured using an ELISA Reader at OD 450 nm, and the serotonin concentration was calculated using a standard curve.

[0021] Figure 1a shows the test results of serotonin changes in subjects before and after taking the composition of an embodiment of the present invention. Figure 1b shows the test results of serotonin changes in subjects before and after taking the composition of the control group. Figure 2 shows the comparison results of serotonin changes in subjects after taking the composition of an embodiment of the present invention and the control group. The values ​​are presented as Mean ± SD. Statistical analysis was performed using paired t-tests, and p < 0.05 was considered statistically significant.

[0022] As can be seen from Figures 1a and 1b, after 2 weeks of taking the composition of the present invention and the control group, the serotonin levels in the blood of the subjects increased significantly compared to before taking the composition (p < 0.001). Furthermore, comparing the change in serotonin levels (i.e., the difference between serotonin levels before and after taking the composition of the present invention) with the control group, the increase in serotonin levels after taking the composition of the present invention was superior to that of the control group, as shown in Figure 2, and the difference was significant (p = 0.0119). Therefore, the composition of the present invention containing five strains of lactic acid bacteria significantly increased serotonin levels more effectively than the control group containing only a single strain of lactic acid bacteria. (II) Effect of taking the composition of the present invention on deep sleep:

[0023] The sleep cycle includes light sleep, deep sleep, and rapid eye movement (REM) sleep. Deep sleep helps the brain rest, repair itself, and restore energy, and also helps strengthen declarative memory. Furthermore, during deep sleep, the pituitary gland secretes growth hormone, which helps body tissues grow and cells regenerate. Sufficient deep sleep is crucial to allow the brain and body enough time to repair; therefore, more deep sleep indicates better sleep quality.

[0024] Before and 2 weeks after taking the compositions of the embodiments of the present invention and the control group, the sleep quality of the subjects was observed using a wearable medical watch sleep analyzer. Figures 3a and 3b show the test results of the proportion of deep sleep and light sleep before and after the subjects took the composition of one embodiment of the present invention, respectively. Figures 4a and 4b show the test results of the proportion of deep sleep and light sleep before and after the subjects took the composition of the control group, respectively. Figures 5 and 6 show the comparison results of the changes in the proportion of deep sleep and light sleep after the subjects took the composition of one embodiment of the present invention and the control group. The values ​​are presented as Mean ± SD. Statistical analysis was performed using paired t-tests, and p < 0.05 was considered statistically significant.

[0025] As can be seen from Figures 3a and 3b, after taking the composition of the embodiments of the present invention, the proportion of light sleep in the subjects decreased, while the proportion of deep sleep increased. Therefore, the subjects' sleep quality was indeed improved after taking the composition of the embodiments of the present invention, and the difference was significant. The subjects also experienced improved sleep quality after taking the composition of the control group, as shown in Figures 4a and 4b.

[0026] In addition, subjects who experienced improved sleep quality after taking the composition of the present invention (i.e., subjects with an increased proportion of deep sleep) were compared with subjects who experienced improved sleep quality after taking the control group composition. The results are shown in Figures 5 and 6. As can be seen from Figure 5, the increase in the proportion of deep sleep after taking the composition of the present invention was better than that of the control group, and the difference was significant (p = 0.0471). Furthermore, as shown in Figure 6, the decrease in the proportion of light sleep was also better than that of the control group, and the difference was significant (p = 0.014). Therefore, the composition of the present invention containing five strains of lactic acid bacteria is better than the control group containing only a single strain of lactic acid bacteria in improving sleep quality. (III) Effect of taking the composition of an embodiment of the present invention on PSQI:

[0027] The Pittsburgh Sleep Quality Index (PSQI) has seven dimensions: sleep quality, sleep latency, sleep duration, sleep efficiency, sleep disturbance, use of sleeping pills, and daytime activity impairment. A higher PSQI score indicates poorer sleep quality.

[0028] Two weeks after taking the composition of the present invention, the sleep quality of the subjects was assessed again using the Pittsburgh Sleep Quality Index (PSQI) questionnaire, and the results are shown in Figure 7. As can be seen from the results in Figure 7, compared with before taking the composition of the present invention, the PSQI was significantly reduced after taking the composition of the present invention for two weeks (p = 0.002), indicating that taking the composition of the present invention significantly improved the sleep quality of the subjects.

[0029] In summary, the compositions of the embodiments of the present invention have physiological activity functions that improve sleep quality. It should be noted that, compared with compositions containing only a single lactic acid bacteria strain, the compositions of the embodiments of the present invention containing five lactic acid bacteria strains can significantly improve sleep quality. Therefore, it can be seen that the combination of lactic acid bacteria strains in the embodiments of the present invention has unexpected effects.

[0030] The above-described embodiments are only for illustrating the technical ideas and features of the present invention. Their purpose is to enable those skilled in the art to understand the content of the present invention and implement it accordingly. They should not be used to limit the patent scope of the present invention. That is, all equivalent changes or modifications made in accordance with the spirit disclosed in the present invention should still be covered within the patent scope of the present invention. [Simplified Explanation of the Diagram]

[0031] Figure 1a shows the test results of changes in serotonin levels in subjects before and after taking the composition of an embodiment of the present invention.

[0032] Figure 1b shows the test results of changes in serotonin levels before and after the subjects took the control group's composition.

[0033] Figure 2 shows the comparison results of serotonin changes after subjects took the composition of an embodiment of the present invention and the control group.

[0034] Figure 3a shows the test results of the percentage of deep sleep in subjects before and after taking the composition of an embodiment of the present invention, using a wearable medical watch sleep analyzer.

[0035] Figure 3b shows the test results of the percentage of light sleep in subjects before and after taking the composition of an embodiment of the present invention, using a wearable medical watch sleep analyzer.

[0036] Figure 4a shows the results of the experiment on the percentage of deep sleep before and after subjects took the control group composition, using a wearable medical watch sleep analyzer.

[0037] Figure 4b shows the results of the experiment on the percentage of light sleep before and after subjects took the control group composition, using a wearable medical watch sleep analyzer.

[0038] Figure 5 shows the comparison results of the change in the proportion of deep sleep after the subjects took the composition of an embodiment of the present invention and the control group.

[0039] Figure 6 shows the comparison results of the change in the proportion of light sleep after the subjects took the composition of an embodiment of the present invention and the control group.

[0040] Figure 7 shows the results of the Pittsburgh Sleep Quality Index (PSQI) test before and after the subjects took the composition of an embodiment of the present invention. [Biomaterial Storage]

[0042] 1. Taiwan, Republic of China, Food Industry Development Institute, 2024 / 12 / 10, BCRC 911254 2. Taiwan, Republic of China, Food Industry Development Institute, 2024 / 12 / 10, BCRC 911251 3. Taiwan, Republic of China, Food Industry Development Institute, 2023 / 02 / 20, BCRC 911172 4. Taiwan, Republic of China, Food Industry Development Institute, 2024 / 12 / 10, BCRC 911252 5. Taiwan, Republic of China, Food Industry Development Institute, 2025 / 01 / 09, BCRC 911262

Claims

1. A composition for improving sleep quality, comprising: a lactic acid bacteria strain having an active effect on improving sleep quality, the lactic acid bacteria strain comprising: isolated *Bifidobacterium breve* BBr-28 strain, accession number BCRC 911254; *Bifidobacterium animalis subsp. lactis* PS23 strain, accession number BCRC 911251; *Bifidobacterium longum subsp. infantis* BR631 strain, accession number BCRC 911172; *Bifidobacterium bifidum* BD-188 strain, accession number BCRC 911252; and *Lactiplantibacillus plantarum* PQQ-1 strain, accession number BCRC 911262, the above strains are all stored at the Food Industry Research and Development Institute, the effective viable count of the lactic acid bacteria strain is equal to or greater than 5×10⁹ CFU / g; and the carrier.

2. The composition for improving sleep quality as described in claim 1, wherein the carrier is a physiologically acceptable excipient or diluent.

3. The composition for improving sleep quality as described in claim 1, wherein the carrier is a pharmaceutically acceptable excipient or diluent.

4. The composition for improving sleep quality as described in claim 1, wherein the dosage form is a tablet, capsule or powder.

5. Use of a lactic acid bacteria strain in the preparation of a composition for improving sleep quality, wherein the composition comprises: a lactic acid bacteria strain having an activity that improves sleep quality, the lactic acid bacteria strain comprising the isolated *Bifidobacterium breve* BBr-28 strain, accession number BCRC 911254; *Bifidobacterium animalis subsp. lactis* PS23 strain, accession number BCRC 911251; *Bifidobacterium longum subsp. infantis* BR631 strain, accession number BCRC 911172; *Bifidobacterium bifidum* BD-188 strain, accession number BCRC 911252; and *Lactiplantibacillus plantarum* PQQ-1 strain, accession number BCRC 911262, the above strains are all stored at the Food Industry Research and Development Institute, the effective viable count of the lactic acid bacteria strain is equal to or greater than 5×10⁹ CFU / g; and the carrier.

6. The use as described in claim 5, wherein the carrier is a physiologically acceptable excipient or diluent.

7. The use as described in claim 5, wherein the carrier is a pharmaceutically acceptable excipient or diluent.

8. For the purposes described in claim 5, the dosage form is tablets, capsules or powder.

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