TWI861750B - Probiotic composition for imrpoving sleep quality and use thereof - Google Patents

Abstract

The present invention relates to a probiotic composition for imrpoving sleep quality and a use thereof. The probiotic composition, which includes Lactobacillus fermentumGKF3, Lactobacillus brevisGKJOY or the combination thereof, can be administered to a healthy or insomnia subject, for improving the sleep quality such as reduction of the sleep onset latency time, increasing of the total sleep time, improvement of the deep sleep, elevated secretion of melatonin and so on, thereby being as an effective ingredient for use in preparation of various compositions.

Description

Probiotic composition for improving sleep quality and its use

The present invention relates to a probiotic composition and its use, in particular to a probiotic composition and its use for improving sleep quality by shortening the time to fall asleep, increasing the total sleep time, improving deep sleep, and enhancing the secretion of melatonin.

A complete sleep cycle consists of two types, rapid eye movement (REM) and non-rapid eye movement (NREM). During REM sleep, the eyes move rapidly, and the activity of brain neurons during this stage is the same as when awake, showing fast, low-voltage desynchronized brain waves. Because REM sleep is physiologically very different from other sleep stages, sleep stages other than REM are called "non-REM sleep" or NREM, which refers to sleep without rapid eye movement. From light to deep, sleep can be divided into the first, second, third, and fourth stages. During the third and fourth stages of NREM sleep, brain activity drops to the lowest level, which is deep sleep. Many physiological processes that are beneficial to the human body occur during this stage, such as the secretion of growth hormone, recovery of energy, and recovery and repair of the body.

Melatonin is a substance found in animals, plants, fungi and bacteria. In animals, melatonin is a hormone responsible for regulating the biological clock and core temperature. Generally, it can be found that the concentration of melatonin in humans is lower during daytime activities; on the contrary, the concentration of melatonin at night is much higher than that during the day. Studies have also found that people with high psychological stress and insomnia have lower melatonin concentrations in their bodies at night than normal people. Currently, melatonin supplements are used clinically to improve sleep.

Probiotics are generally defined as bacteria that are beneficial to human health, can reproduce in the human intestine and are non-pathogenic. Usually probiotics can regulate the body constitution, change the metabolic pathways of some parts of the human body, and improve the overall gastrointestinal tract, metabolic capacity and other functions.

In recent years, many studies have suggested that gut microorganisms have the potential to improve the host's cognition, memory, mood, and spirit through the brain-gut axis. Among them, the majority of studies are on lactic acid bacteria. For example, a study found that Lactiplantibacillus plantarum can improve anxiety-like or depressive behaviors in mice. Other studies have found that Lactobacillus johnsonii can alleviate the memory loss of mice caused by restraint stress. Other studies have found that depression, memory, mood, etc. are related to sleep quality.

However, current research has not yet examined whether other probiotics can improve the sleep quality of people with insomnia. There is an urgent need to develop a probiotic composition that can improve sleep quality and its use.

Therefore, one aspect of the present invention is to provide a probiotic composition for improving sleep quality, which comprises Lactobacillus fermentum strain GKF3, Lactobacillus brevis strain GKJOY or a combination thereof.

Secondly, another aspect of the present invention is to provide a probiotic for use as a composition for improving sleep quality, including administering an oral composition containing an effective dose of probiotics to a subject with insomnia for a continuous period of time, and the probiotic contains the above-mentioned strain as an active ingredient.

According to the above aspects of the present invention, a probiotic composition for improving sleep quality is proposed. In one embodiment, the probiotic composition for improving sleep quality comprises fermented lactobacillus ( Lactobacillus fermentum ) strain GKF3, short lactic acid bacteria ( Lactobacillus brevis ) strain GKJOY or a combination thereof, fermented lactobacillus strain GKF3 was deposited at the Bioresource Conservation and Research Center of the Food Industry Development Research Institute (BCRC, No. 331, Food Road, Hsinchu, Taiwan) on February 12, 2018, with a deposit number of BCRC 910824, and short lactic acid bacteria strain GKJOY was deposited at BCRC on July 18, 2019, with a deposit number of BCRC 910920.

In the above embodiments, the aforementioned probiotic composition may be, for example, an oral composition, such as a pharmaceutical composition or a food composition. In the above examples, the aforementioned probiotic composition further comprises a pharmaceutically and food-acceptable carrier, excipient and/or additive.

In the above embodiments, the dosage form of the aforementioned probiotic composition may include, for example, powder, tablet, granule, suppository, microcapsule, ampoule, liquid spray or plug.

In the above embodiment, the aforementioned improvement of sleep quality includes shortening the time to fall asleep, increasing the total sleep time, and improving the deep sleep of insomnia subjects.

According to another aspect of the present invention, a use of probiotics in preparing a composition for improving sleep quality is proposed, including administering an oral composition containing an effective dose of probiotics to a subject for at least 14 consecutive days, and the probiotics contain fermentative lactobacillus strain GKF3 (BCRC 910824), breve lactobacillus strain GKJOY (BCRC 910920) or a combination thereof as an active ingredient.

In the above embodiment, the aforementioned subject may be, for example, an adult, and the effective dose of the aforementioned probiotics for the subject may be, for example, 0.2g/50~60kg body weight/day to 3g/50~60kg body weight/day.

In the above embodiment, the aforementioned subject may be, for example, a rat, and the effective dose of the aforementioned probiotics for the subject may be, for example, 50 mg/300 g body weight/day to 300 mg/300 g body weight/day. In one example, the aforementioned rat may be, for example, a healthy rat or an insomnia rat.

Application The above-mentioned probiotic composition for improving sleep quality and its use of the present invention, which includes fermented lactobacillus strain GKF3, short lactic acid bacteria strain GKJOY or a combination thereof, is administered to insomnia subjects for a continuous period of time. It can effectively shorten the time to fall asleep, increase the total sleep time, enhance deep sleep, increase melatonin secretion, etc., and then be used as an effective ingredient and used to prepare various compositions.

It is understood that the foregoing general description and the following detailed description are merely illustrative and are intended to provide further explanation of the claimed invention.

In order to make the above and other purposes, features, advantages and embodiments of the present invention more clearly understandable, the attached figures are described in detail as follows: [Figure 1] is a bar graph showing the melatonin concentration in the saliva of the subject before falling asleep after the short lactic acid bacteria strain GKJOY according to one embodiment of the present invention was administered to the subject.

[Figure 2] is a bar graph showing the melatonin concentration in the saliva of the subjects before falling asleep after the fermented lactobacillus strain GKF3 according to one embodiment of the present invention was administered to the subjects.

[Figure 3] is a curve graph and a bar graph showing the percentage of non-rapid eye movement sleep time of the subjects after the short lactic acid bacteria strain GKJOY according to one embodiment of the present invention was administered to the subjects.

[Figure 4] is a curve chart and a bar chart showing the percentage of awake time of the subjects after the short lactic acid bacteria strain GKJOY according to one embodiment of the present invention was administered to the subjects.

[Figure 5] is a curve chart and a bar chart showing the percentage of non-rapid eye movement sleep time of insomnia subjects after the short lactic acid bacteria strain GKJOY according to one embodiment of the present invention was administered to the insomnia subjects.

If the definition or use of a term in a reference is inconsistent or contrary to the definition of that term here, the definition of that term here shall apply rather than the definition of that term in the reference. Secondly, unless the context otherwise requires, a singular term may include the plural and a plural term may include the singular.

As mentioned above, the present invention provides a probiotic composition for improving sleep quality and its use. After being administered to healthy or insomniac subjects for a continuous period of time, it can effectively improve sleep quality, such as shortening the time to fall asleep, increasing the total sleep time, improving deep sleep, and enhancing melatonin secretion.

In one embodiment, the "probiotics" referred to herein may refer to probiotics of different species or genera, for example, Lactobacillus fermentum strain GKF3, Lactobacillus brevis strain GKJOY or a combination thereof, wherein Lactobacillus fermentum strain GKF3 was deposited at the Bioresource Conservation and Research Center of the Food Industry Development Research Institute (BCRC, No. 331, Food Road, Hsinchu, Taiwan) on February 12, 2018, with the deposit number BCRC 910824, and Lactobacillus brevis strain GKJOY was deposited at BCRC on July 18, 2019, with the deposit number BCRC 910920. It should be noted that the present invention requires the use of probiotics of specific strains so that the obtained probiotic composition can exert the effect of improving sleep quality. If the probiotics are not selected from the above-mentioned specific strains, or if the strains are increased or decreased, or if part or all of the probiotics are replaced with other strains of the same species, the effect of improving sleep quality cannot be expected.

Compared to healthy subjects, the "insomnia subjects" referred to herein are defined as having difficulty falling asleep (i.e., taking too long to fall asleep, or being unable to fall asleep again after waking up in the morning, or insufficient deep sleep) and/or having difficulty staying asleep (i.e., the total sleep time is too short, or the total wakefulness time is too long). It is generally recognized that the sleep cycle of organisms is related to the light-dark cycle, and changes in the light-dark cycle can affect the melatonin secretion of humans or animals, so the melatonin secretion can also be used as one of the indicators of sleep quality. In some embodiments, after falling asleep, the insomnia subjects have insufficient melatonin secretion. In some specific examples, the aforementioned insomnia subjects can be induced to develop insomnia using a learning method, such as administering caffeine 30 minutes before the light cycle to induce insomnia in the subjects, so as to facilitate the subsequent evaluation of the effect of the probiotic composition on improving sleep quality.

The evaluation items of "sleep quality" referred to herein may include but are not limited to the time to fall asleep, total sleep time, deep sleep, melatonin secretion, etc. Compared with healthy subjects, insomnia subjects have longer time to fall asleep, shorter total sleep time, poorer deep sleep, and less melatonin secretion. Therefore, in one embodiment, administering an oral composition containing an effective amount of probiotics to insomnia subjects for at least 14 consecutive days can significantly improve the sleep quality of insomnia subjects, including but not limited to shortening the time to fall asleep, increasing the total sleep time, improving deep sleep, and enhancing melatonin secretion, etc.

In one embodiment, the content of each strain of the above-mentioned probiotic composition is not particularly limited. In one example, the probiotic composition may be composed only of fermented lactobacillus GKF3 (BCRC 910824) or short lactobacillus GKJOY (BCRC 910920). In another example, the probiotic composition includes fermented lactobacillus GKF3 (BCRC 910824) and short lactobacillus GKJOY (BCRC 910920), and their weight (mg) ratio or bacterial count (CFU) ratio may be, for example, 1:1. However, in other examples, the weight (mg) ratio or bacterial count (CFU) ratio of the above-mentioned fermented lactobacillus GKF3 (BCRC 910824) and short lactobacillus GKJOY (BCRC 910920) may also use a ratio other than 1:1, such as (1~4):1 or 1:(1~4).

When used, the probiotic composition may be, for example, an oral composition, such as a pharmaceutical composition or a food composition. The aforementioned probiotic usage may include, but is not limited to, fermentation liquid, bacterial sludge (or bacterial pellet), supernatant, freeze-dried powder and other unknown and inseparable active ingredients. In the above embodiments, the aforementioned fermentation liquid refers to the product including bacterial cells and culture liquid. The aforementioned bacterial sludge refers to the product after the fermentation liquid is depleted of supernatant. The aforementioned supernatant refers to the product after the fermentation liquid is depleted of bacterial sludge. The aforementioned freeze-dried powder refers to the freeze-dried powder prepared from the fermentation liquid, bacterial sludge and/or supernatant. It is understandable that the aforementioned fermentation liquid, bacterial sludge, supernatant and freeze-dried powder are only preliminarily processed and still contain other unknown and inseparable active ingredients.

Examples of the aforementioned pharmaceutical compositions may include but are not limited to medicines. Examples of probiotics applied to food compositions may include but are not limited to general foods, health foods, beverages, nutritional supplements, dairy products or feeds, etc. In the aforementioned examples of oral compositions, the probiotic composition may optionally include pharmaceutically and food-acceptable carriers, excipients and/or additives. In other examples, the dosage form of the probiotic composition may include but is not limited to powders, tablets, granules, suppositories, microcapsules, ampoules, liquid sprays or plugs. In the above examples, the probiotic composition may also optionally include excipients, preservatives, diluents, fillers, absorption enhancers, sweeteners or any combination thereof.

When the above-mentioned probiotics are used to prepare a composition for improving sleep quality, an oral composition containing an effective dose of probiotics can be administered to the subject for a continuous period of time. The above-mentioned effective dose depends on the subject and is not particularly limited, but the above-mentioned probiotics administered include the strain and other inseparable active ingredients (such as fermentation products, metabolites, components of the culture medium, etc. of the strain). For example, when the subject is an adult, the effective dose of probiotics for adults can be, for example, 0.2g/50~60kg body weight/day to 3g/50~60kg body weight/day, but 0.6g/50~60kg body weight/day to 2g/50~60kg body weight/day, and about 0.8g/50~60kg body weight/day to 1g/50~60kg body weight/day is more preferred. In another example, when the subject is a rat (e.g., a healthy rat or an insomnia rat), the effective dose of the probiotics for the subject may be, for example, 50 mg/300 g body weight/day to 300 mg/300 g body weight/day, preferably 100 mg/300 g body weight/day to 200 mg/300 g body weight/day, and more preferably about 100 mg/300 g body weight/day. In other examples, the aforementioned administration period is not particularly limited, and may be, for example, continuous for at least 14 days, or may be longer or shorter. In general, the number of bacteria per gram (g) of the aforementioned probiotics may be, for example, 1×10 ⁹ to 1×10 ¹¹ colony forming units (CFU).

It is understood that the following specific strains, specific formulations, specific dosages, specific detection methods, viewpoints, examples and embodiments are only for illustration and are not intended to be limiting conditions of the present invention. The main features of the present invention can be used in various embodiments without departing from the spirit and scope of the present invention. Therefore, those with ordinary knowledge in the technical field to which the present invention belongs can easily determine the necessary technical features of the present case without departing from the spirit and scope of the present invention, and make various changes and modifications to the present invention to apply it to different uses and conditions.

Example 1

1.1 Source of strains

The strains used in this embodiment include a specific combination of probiotics, including fermented lactobacillus ( Lactobacillus fermentum ) strain GKF3, short lactic acid bacteria ( Lactobacillus brevis ) strain GKJOY or a combination thereof. Fermented lactobacillus strain GKF3 was deposited at the Bioresource Conservation and Research Center of the Food Industry Development Research Institute (BCRC, No. 331, Food Road, Hsinchu, Taiwan) on February 12, 2018, with the deposit number BCRC 910824, and short lactic acid bacteria strain GKJOY was deposited at BCRC on July 18, 2019, with the deposit number BCRC 910920.

The fermentative lactobacillus strain GKF3 and the short lactic acid bacteria strain GKJOY can be isolated from sour kimchi and Taiwanese kimchi. For example, the fermentative lactobacillus strain GKF3 is isolated from Taiwanese kimchi, and the short lactic acid bacteria strain GKJOY is isolated from sour kimchi.

In some examples, the fermentative lactobacillus strain GKF3 and the short lactic acid bacteria strain GKJOY are inoculated on a solid culture medium to activate the strains. In a preferred embodiment, the solid culture medium is commercially available MRS (deMan, Rogosa and Sharpe) agar. After the colony is generated, a single colony is picked out and inoculated into a liquid culture medium for liquid culture. In a preferred embodiment, the strain culture temperature is 25°C to 40°C. In a preferred embodiment, the liquid culture time is 16 to 24 hours. In a preferred embodiment, the liquid culture medium is MRS liquid culture medium. After the liquid culture is completed, fermentation culture is performed. In a preferred embodiment, the formula of the fermentation medium is shown in Table 1 below.

1.2 Preparation of freeze-dried powder

After the fermentation lactobacillus strain GKF3 and the short lactic acid bacteria strain GKJOY are grown by fermentation culture, the fermentation liquid containing the bacteria and the culture liquid is collected and centrifuged at a rate of 1000 rpm to 15000 rpm to obtain bacterial mud. The obtained bacterial mud is mixed with a preservative (i.e., 6% to 50% by weight of skimmed milk powder) and then freeze-dried. The freeze-drying is pre-frozen in a gradient setting mode, and is stored at 20°C to 0°C for 1 to 4 hours, then stored at 0°C to -20°C for 4 to 8 hours, and then stored at -196°C to -30°C for more than 8 hours. In a preferred embodiment, the freeze-drying temperature and time are stored at -40°C for 2 hours, then at -20°C for 2 hours, then at 0°C for 2 hours, and finally at 20°C for more than about 10 hours. Afterwards, the freeze-dried powder obtained above is stored at a low temperature. In another preferred embodiment, the temperature of the low temperature storage is -30°C to 4°C. The stored freeze-dried powder can be used as the raw material of the probiotic composition to be administered to the test animals in the following animal experiments.

Example 2: Clinical trial to evaluate the effect of the probiotic strain in Example 1 on improving sleep quality

2.1 Inclusion and grouping

The following example is a single-blind, randomized before-after controlled clinical study. A total of 6 healthy adult women were recruited as subjects and randomly divided into two groups, one group was oral strain GKJOY, and the other group was oral strain GKF3, with 3 people in each group. The changes before and after the subjects took the strains orally were used as the results for comparison.

2.2 Experimental design

1 mL of saliva samples were collected from the subjects before and after each second oral administration of the strain, and a total of 4 saliva samples were collected, and the samples were kept frozen until analysis. The interval between each saliva collection was at least one day, and the first saliva collection day was the first day of the test. The subjects began to take the test strain orally the day after the second saliva collection before going to bed (the third day of the test), with a daily dose of 0.8 grams, and continued to take the test strain orally for 14 consecutive days. Saliva samples were collected from each person before going to bed (21:30 to 23:00) on the 1st, 2nd, 15th, and 16th days of the test, and the saliva samples were tested on the 15th and 16th days of the test, which was about two weeks after the test strain was taken orally. The concentration of melatonin was analyzed from the subjects' saliva samples to assess sleep quality.

2.3 Test the melatonin concentration in saliva before bedtime (pg/mL)

The enzyme-linked immunosorbent assay (ELISA, also known as enzyme immunoassay) was used in conjunction with the commercially available human melatonin kit [Human MT (Melatonin) kit] to analyze the melatonin concentration (pg/mL) in the subjects' saliva according to the instructions provided by the manufacturer. Generally speaking, the melatonin concentration in the subjects' saliva when awake is less than 5pg/mL, and the melatonin concentration in the saliva before going to bed is greater than 10pg/mL. The results are shown in Figures 1 and 2.

Please refer to Figure 1, which shows the melatonin concentration in the saliva of the subjects before going to sleep after the short lactic acid bacteria strain GKJOY according to an embodiment of the present invention was administered to the subjects. Compared with the melatonin concentration in the saliva before going to sleep measured before taking the strain GKJOY (pre-test, 1.34±0.038pg/mL), the melatonin concentration in the saliva before going to sleep after taking the strain GKJOY (post-test, 1.40±0.046pg/mL) increased significantly. From the results of the average melatonin concentration in the saliva of the subjects before going to sleep, it can be found that most people are in a mentally active state before going to bed, and it is difficult to fall asleep, and it takes a long time to help fall asleep. However, this example proves that the melatonin concentration (pg/mL) in the saliva of the three subjects before bedtime about 14 days after oral administration of strain GKJOY (post-test) is greater than the saliva melatonin concentration before oral administration of strain GKJOY (pre-test), proving that strain GKJOY in Example 1 helps to increase the melatonin concentration in saliva before bedtime and helps the subjects fall asleep.

Please refer to Figure 2, which shows a bar graph of the melatonin concentration in the saliva of the subjects before falling asleep after the fermented lactobacillus strain GKF3 according to an embodiment of the present invention was administered to the subjects. Compared with the melatonin concentration in the saliva before falling asleep measured before taking the drug (pre-test, 1.35±0.069pg/mL), the melatonin concentration in the saliva of the other three subjects before falling asleep about 14 days after oral administration of the strain GKF3 (post-test, 1.39±0.013pg/mL) was greater than the saliva melatonin concentration before oral administration of the strain GKF3 (pre-test), proving that the strain GKF3 of Example 1 also helps to increase the melatonin concentration in the saliva before falling asleep and helps the subjects fall asleep.

Example 3: Using animal test model to evaluate the effect of probiotics in improving sleep

Since both strain GKF3 and strain GKJOY have the effect of improving sleep, Example 3 uses strain GKJOY as an example and selects male Sprague-Dawley (SD) rats to evaluate the effect of probiotics in improving sleep.

SD rats were housed in 28×22×40 cm ³ cages with an average temperature of approximately 22±2°C and an average humidity of 50±5%. The rats were kept in a 12-hour light and dark cycle and provided with sufficient drinking water and standard feed. All animal experimental procedures were reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) of National Taiwan University, and obtained the IACUC approval number NTU-109-EL-00133.

Animal experiments are divided into two experimental designs. Experimental design 1 divides the experimental animals into a blank control group (Vehicle) and a test strain GKJOY group (abbreviated as GKJOY). Experimental design 2 divides the experimental animals into a blank control group (Vehicle), a caffeine-induced group (abbreviated as Caffeine), and a caffeine plus test strain GKJOY group (abbreviated as Caffeine+GKJOY).

3.1 Animal trial design I: Long-term effects of probiotics on sleep

Rats in the blank control group (Vehicle) were orally administered sterilized water daily, while rats in the test strain GKJOY group (GKJOY) were administered with lactic acid bacteria GKJOY and its active ingredients for 14 consecutive days, with the administration of the test substance as the first day of the experiment. The dosage of lactic acid bacteria GKJOY and its active ingredients in the experimental group was 100 mg/kg, and the administration time was 90 minutes before entering the light cycle. Because rats are nocturnal animals, sleep observations of this species mostly occur within the 5-light cycle (sleep period), and sleep-related indicators of rats were recorded on the 1st, 7th, and 14th days of the experiment. The single recording time was about 24 hours, and the analysis items included the rapid eye movement (REM) period and the proportion of awake time.

3.2 Animal experiment design II: Effects of probiotics on caffeine-induced insomnia

Except for the rats in the blank control group (Vehicle), the test animals in the other groups were injected intraperitoneally (i.p.) with 100 mg/kg caffeine 30 minutes before entering the light cycle to induce insomnia in the rats. The rats in the blank control group (Vehicle) and the caffeine induced group (Caffeine) were orally administered with sterile water 90 minutes before entering the light cycle; the rats in the caffeine plus test strain GKJOY group (Caffeine+GKJOY) were orally administered with strain GKJOY (100 mg/kg) 90 minutes before entering the light cycle. The proportion of non-rapid eye movement (deep sleep) of normal mice and insomnia rats was recorded after entering the light cycle.

3.3 Statistics

The values described in this article are expressed as mean ± standard deviation (mean ± SD) and analyzed using one-way ANOVA and commercially available software. P < 0.05 was considered statistically significant, with asterisks * representing p < 0.05 and asterisks ** representing p < 0.01.

4. Probiotics shorten the sleep time of healthy rats

Please refer to Figure 3, which is a curve graph (large picture) and a bar graph (small picture) showing the percentage of non-rapid eye movement sleep time (% of recording time) of the subject (rats) after the short lactic acid bacteria strain GKJOY according to one embodiment of the present invention was administered to the subject.

The results in Figure 3 show that compared with the blank control group (Vehicle), the time taken by rats that took oral administration of strain GKJOY to enter the non-rapid eye movement phase (sleep) during the light cycle (sleep period) is shorter. Secondly, the sleep time of rats taking oral administration of strain GKJOY on the 1st, 7th, and 14th days shows that the longer the oral administration of strain GKJOY, the shorter the time required for rats to fall asleep during the light cycle (sleep period). For example, on the 14th day of oral administration of strain GKJOY, the proportion of rats that took oral administration of strain GKJOY in the non-rapid eye movement phase (sleep) was significantly higher than that of the blank control group (Vehicle) (p<0.01), proving that strain GKJOY has the effect of helping to fall asleep.

5. Probiotics increase total sleep time in healthy rats

Please refer to Figure 4, which shows a curve graph (large graph) and a bar graph (small graph) of the percentage of awake time of the subject after the short lactic acid bacteria strain GKJOY according to one embodiment of the present invention was administered to the subject.

The results in Figure 4 show that compared with the blank control group (Vehicle), the rats that took the strain GKJOY orally had a lower percentage of total wakefulness time (% of recording time) during the light cycle (sleep period), that is, the total sleep time increased. As the time of oral administration of strain GKJOY increases, the rats’ wakefulness ratio during the light cycle (sleep period) decreases. For example, the total sleep time of rats that took the strain GKJOY orally for 14 days was significantly higher than that of the blank control group (Vehicle) (p<0.01), proving that strain GKJOY has the effect of increasing total sleep time.

6. Probiotics improve deep sleep in insomnia rats

Please refer to Figure 5, which shows the curve graph (large graph) and bar graph (small graph) of the percentage of non-rapid eye movement sleep time of insomnia subjects (insomnia rats) after the short lactic acid bacteria strain GKJOY according to one embodiment of the present invention was administered to the insomnia subjects.

The results in Figure 5 show that compared with the blank control group (Vehicle), the percentage of non-rapid eye movement sleep (deep sleep) time (% of recording time) of insomnia rats induced by caffeine (Caffeine) was significantly reduced, indicating that the rats were not easy to fall asleep. On the contrary, insomnia rats that took the strain GKJOY orally (Caffeine+GKJOY) could significantly improve the phenomenon of reduced deep sleep, proving that the strain GKJOY has the effect of enhancing the deep sleep of insomnia rats.

In summary, the present invention uses specific strains, specific formulas, specific dosages, specific detection methods, or specific evaluation methods only to illustrate the probiotic composition for improving sleep quality and its use. However, those with ordinary knowledge in the art to which the present invention belongs should understand that within the spirit and scope of the present invention, the use of strain GKF3, strain GKJOY or a combination thereof, other formulas, other dosages, other detection methods, or other evaluation methods can also be used for the probiotic composition for improving sleep quality and its use, and are not limited to the above. For example, the probiotic composition may include fermented lactobacillus strain GKF3 and Lactobacillus brevis . The probiotic composition may also be a pharmaceutical composition or a food composition, and may optionally include pharmaceutically and food-acceptable carriers, excipients and/or additives, and may be prepared in the form of powders, tablets, granules, suppositories, microcapsules, ampoules, liquid sprays or plugs.

According to the above embodiments, the probiotic composition for improving sleep quality of the present invention has the advantage that the probiotic composition comprises fermented lactobacillus GKF3, breve lactobacillus GKJOY or a combination thereof, and can effectively improve sleep quality, such as shortening the time to fall asleep, increasing the total sleep time, improving deep sleep, and enhancing melatonin secretion, by being administered to healthy or insomniac subjects for a continuous period of time. In the future, it can be used as an active ingredient and used to prepare various compositions.

Although the present invention has been disclosed above with several specific embodiments, other embodiments are also possible. Therefore, the spirit and scope of the appended claims of the present invention should not be limited to the embodiments contained herein.

【Biological material storage】 Domestic storage information (please note the order of storage institution, date, and number)

The strain GKF3 of Lactobacillus fermentum was deposited at the Bioresource Conservation and Research Center of the Food Industry Development Research Institute, No. 331, Food Road, Hsinchu, Taiwan, on February 12, 2018, with the deposit number BCRC 910824, and was confirmed alive on March 1, 2018. The strain GKJOY of Lactobacillus brevis was deposited at the Bioresource Conservation and Research Center of the Food Industry Development Research Institute, No. 331, Food Road, Hsinchu, Taiwan, on July 18, 2019, with the deposit number BCRC 910920, and was confirmed alive on July 26, 2019.

Overseas storage information (please note the order of storage country, institution, date and number)

without

Claims (11)

A probiotic composition for improving sleep quality, comprising Lactobacillus fermentum strain GKF3, Lactobacillus brevis strain GKJOY or a combination thereof, wherein the Lactobacillus fermentum strain GKF3 was deposited at the Bioresource Conservation and Research Center of the Food Industry Development Research Institute (BCRC, No. 331, Food Road, Hsinchu, Taiwan) on February 12, 2018, with the deposit number BCRC 910824, and the Lactobacillus brevis strain GKJOY was deposited at the BCRC on July 18, 2019, with the deposit number BCRC 910920. A probiotic composition for improving sleep quality as described in claim 1, wherein the probiotic composition is an oral composition. A probiotic composition for improving sleep quality as described in claim 2, wherein the probiotic composition is a pharmaceutical composition or a food composition. The probiotic composition for improving sleep quality as described in claim 2 further comprises a pharmaceutically and food-acceptable carrier, formulator and/or additive. A probiotic composition for improving sleep quality as described in claim 1, wherein one dosage form of the probiotic composition includes powder, tablet, granule, suppository, microcapsule, ampoule, liquid spray or plug. The probiotic composition for improving sleep quality as described in claim 1, wherein the improvement of sleep quality includes shortening the time to fall asleep, increasing the total sleep time, improving deep sleep and increasing the secretion of melatonin. A use of a probiotic in preparing a composition for improving sleep quality, comprising administering an oral composition containing an effective dose of the probiotic to a subject for at least 14 consecutive days, and the probiotic contains fermentative lactobacillus strain GKF3 (BCRC 910824), breve lactobacillus strain GKJOY (BCRC 910920) or a combination thereof as an active ingredient. The use of probiotics as described in claim 7 in preparing a composition for improving sleep quality, wherein the improvement of sleep quality includes shortening the time to fall asleep, increasing the total sleep time, and improving the deep sleep of insomniac subjects. The use of probiotics in preparing a composition for improving sleep quality as described in claim 7, wherein the subject is an adult, and the effective dose of the probiotics administered to the subject is 0.2g/50~60kg body weight/day to 3g/50~60kg body weight/day. The use of probiotics in preparing a composition for improving sleep quality as described in claim 7, wherein the subject is a rat, and the effective dose of the probiotics administered to the subject is 50 mg/300 g body weight/day to 300 mg/300 g body weight/day. The use of probiotics as described in claim 10 in preparing a composition for improving sleep quality, wherein the rat is a healthy rat or an insomnia rat.