ghrelin secretion promoters

Abstract

The present invention aims to provide a novel ghrelin secretion enhancer that is easily accessible and easy to consume, can stimulate the secretion of ghrelin present in the human body, and is safe and has no side effects. By ingesting specific lactic acid bacteria themselves or fermented milk produced using them as a ghrelin secretion enhancer, poor appetite can be significantly alleviated in an easily accessible, easy, and safe manner.

Description

[0001] This application is a divisional application of the invention patent application filed on July 13, 2017, with application number 201780043060.3 and invention title "Hunger Gluttony Secretion Promoter". Technical Field

[0002] This invention relates to ghrelin secretion promoters. Background Technology

[0003] Ghrelin is a peptide hormone primarily secreted in the stomach. It has been reported to have effects such as increased appetite, regulation of gastrointestinal motility, growth hormone promotion, regulation of energy metabolism, increased cardiac function, and anti-diabetic effects. Therefore, promoting ghrelin secretion can improve loss of appetite, making substances that promote ghrelin secretion very useful. To date, reports have documented the use of milk components, plant extracts, or traditional Chinese medicine to promote ghrelin secretion, as well as the use of lactic acid bacteria. Therefore, it is desirable to obtain the ghrelin-promoting effect through simpler methods, such as consuming common foods prepared with lactic acid bacteria, without the use of pharmaceuticals.

[0004] As a previous example, Japanese Patent Publication No. 2009-524640 (Patent Document 1) describes the use of Lactobacillus acidophilus, Lactobacillus salivarius, and Lactobacillus curvatus to regulate ghrelin secretion. Japanese Patent Publication No. 2015-127338 (Patent Document 2) describes a composition containing lactic acid bacteria of the Lactobacillus genus and ghrelin. Japanese Patent Publication Nos. 2015-205829 (Patent Document 3) and 2015-205830 (Patent Document 4) describe ghrelin secretion promoters using skim milk or skim milk powder as active ingredients. However, these documents do not describe or suggest that lactic acid bacteria, particularly Lactobacillus gasseri, can promote ghrelin secretion and improve loss of appetite.

[0005] Existing technical documents

[0006] Patent documents

[0007] Patent Document 1: Japanese Patent Publication No. 2009-524640

[0008] Patent Document 2: Japanese Patent Application Publication No. 2015-127338

[0009] Patent Document 3: Japanese Patent Application Publication No. 2015-205829

[0010] Patent Document 4: Japanese Patent Application Publication No. 2015-205830

[0011] Summary of the invention

[0012] The technical problem that the invention aims to solve

[0013] To date, in order to use ghrelin to improve loss of appetite, it is necessary to semi-force the intake (giving) of ghrelin or a known substance that induces the secretion of ghrelin, which presents physical and / or psychological barriers in terms of intake.

[0014] Against this backdrop, the present invention aims to provide a novel ghrelin secretion promoter that is readily ingested, can exert the ghrelin secretion effect present in the human body, and is safe and without side effects.

[0015] Technical solutions adopted to solve technical problems

[0016] To address the aforementioned issues, the inventors focused on fermented milk as an easily accessible and readily consumed food, and conducted a detailed investigation into the functionality of lactic acid bacteria used in the preparation of fermented milk. As a result, it was discovered that the specified lactic acid bacteria and the fermented milk prepared using them exhibit a significant ghrelin secretion effect, thus completing this invention.

[0017] That is, the present invention is as follows.

[0018] [1] A ghrelin secretion promoter, characterized in that it contains lactic acid bacteria of the genus Lactobacillus as an active ingredient.

[0019] [2] The ghrelin secretion promoter according to [1] is characterized in that the lactobacillus is Lactobacillus gasseri.

[0020] [3] The ghrelin secretion promoter according to [1] or [2] is characterized in that the lactobacillus lactobacillus is Lactobacillus gasseri OLL2716 (FERM BP-6999).

[0021] [4] The ghrelin secretion promoter according to any one of [1] to [3] is characterized in that it is used for Helicobacter pylori-negative individuals.

[0022] [5] A fermented milk, wherein it comprises the ghrelin secretion promoter described in any one of [1] to [4].

[0023] In addition, the present invention also includes the following inventions.

[0024] [6] A method for promoting ghrelin secretion, comprising the step of ingesting fermented milk prepared using lactic acid bacteria of the genus Lactobacillus.

[0025] [7] The method for promoting ghrelin secretion according to [6], wherein the lactic acid bacteria of the genus Lactobacillus is Lactobacillus gasseri.

[0026] [8] The method for promoting ghrelin secretion according to [6] or [7], wherein the Lactobacillus gasseri is Lactobacillus gasseri OLL2716 (FERM BP-6999).

[0027] [9] Use of lactic acid bacteria of the genus Lactobacillus in the manufacture of ghrelin secretion promoters.

[0028]

[10] According to the use described in [9], the lactic acid bacteria of the genus Lactobacillus is Lactobacillus gasseri.

[0029]

[11] According to the use described in [9] or

[10] , wherein the Lactobacillus gasseri is Lactobacillus gasseri OLL2716 (FERM BP-6999).

[0030]

[12] A lactic acid bacterium of the genus Lactobacillus, wherein the lactic acid bacterium is used in the manufacture of a ghrelin secretion promoter.

[0031]

[13] Lactobacillus gasseri OLL2716 (FERM BP-6999), a lactic acid bacterium of the genus Lactobacillus, is used in the manufacture of ghrelin secretion promoters.

[0032] The effects of the invention

[0033] By employing this invention, the ingestion of prescribed lactic acid bacteria or the use of fermented milk prepared therefrom can promote ghrelin secretion, significantly improving loss of appetite. In particular, fermented milk is a widely known and readily available food, easily and safely consumed, thus providing substantial assistance in improving loss of appetite.

[0034] Brief description of the attached diagram

[0035] Figure 1 This graph shows the results of comparing the serum ghrelin concentrations in the control group and the LG21 yogurt group in Example 1. The vertical lines represent the standard deviation within each group. There was a statistically significant difference between the control group and the LG21 yogurt group at a significance level of less than 5%.

[0036] Figure 2 This graph shows the results of comparing the ghrelin gene expression levels in gastric tissues of the control group and the LG21 yogurt group in Example 1. The vertical lines represent the standard deviation within each group. There was a statistically significant difference between the control group and the LG21 yogurt group at a significance level of less than 5%.

[0037] Figure 3This graph shows the results of comparing the ghrelin gene expression levels in gastric tissues of the control group, 1 hour after administration of LG21 live bacteria, and 2 hours after administration of LG21 live bacteria in Example 2. The vertical lines represent the standard deviation within each group. There were statistically significant differences between the control group and 2 hours after administration of LG21 live bacteria, and between 1 hour and 2 hours after administration of LG21 live bacteria, at a significance level of less than 5%.

[0038] Methods of implementing the invention

[0039] In this invention, lactic acid bacteria can be used directly. The term "lactic acid bacteria" in this invention refers to all bacteria taxonomically recognized as lactic acid bacteria, without limitation on species or strains. Furthermore, lactic acid bacteria are also classified according to their origin as plant-based or animal-based; the lactic acid bacteria of this invention can be of either plant or animal origin. In this invention, since they are more widely known in fermented milk such as yogurt, it is preferable to use one or more strains of lactic acid bacteria selected from the genus *Lactobacillus*. Moreover, it is more preferable that the *Lactobacillus* genus lactic acid bacteria is *Lactobacillus gasseri*, and even more preferably *Lactobacillus gasseri* OLL2716 (FERM BP-6999).

[0040] Here, “Lactobacillus gasseri OLL2716” was internationally deposited on May 24, 1999 (original deposit date) at the Patent Biology Collection Center of the National Institute of Advanced Industrial Science and Technology (NIAS), Japan Post Code 305-8566, 1-1-1-Chuo-6, Higashi, Tsukuba City, Ibaraki Prefecture, Japan, under accession number FERM BP-6999, based on the Budapest Treaty. This strain was originally deposited domestically under the domestic accession number P-17399 on May 24, 1999 (original accession), and was transferred to international accession based on the Budapest Treaty on January 14, 2000.

[0041] As recorded in Budapest Notification No. 282 (http: / / www.wipo.int / treaties / en / notifications / budapest / treat y_budapest_282.html), the National Institute of Advanced Industrial Science and Technology, International Patent Biological Collection Center (IPOD, NITE) inherited the patent microbial collection business from the aforementioned National Institute of Advanced Industrial Science and Technology, Patent Biological Collection Center (IPOD, AIST), and is therefore currently deposited at the National Institute of Advanced Industrial Science and Technology, International Patent Biological Collection Center (IPOD, NITE) (Room 120, 2-5-8 Kazusa-Kamazutari, Kisarazu City, Chiba Prefecture, Japan Postal Code 292-0818) (FERM BP-6999).

[0042] Furthermore, in another embodiment of the present invention, fermented milk can be prepared using lactic acid bacteria. In this invention, compared to the case of ingesting lactic acid bacteria, fermented milk provides a moderately refreshing sour taste, allowing for the easy and effortless consumption of the *Lactobacillus* species of lactic acid bacteria. Examples of methods for preparing fermented milk include sterilizing raw milk, cooling it, adding a lactic acid bacteria starter culture, and fermenting at a specified lactic acid level, temperature, and time. Examples of lactic acid levels include, for example, 0.6–1.2% by mass. Examples of fermentation temperatures include, for example, 40–45°C. Examples of fermentation times include, for example, 2–12 hours.

[0043] In this specification, "fermented milk" refers to products obtained by fermenting milk, including, for example, "fermented milk," "lactic acid bacteria beverages," "milk beverages," and "natural cheeses" as defined by regulations concerning the composition standards of milk and dairy products (Milk, etc. Regulations), but is not limited to these. For example, fermented milk refers to "fermented milk" as defined by the Milk, etc. This means products obtained by fermenting raw milk, cow's milk, special cow's milk, raw goat's milk, pasteurized goat's milk, raw sheep's milk, ingredient-adjusted milk, low-fat milk, non-fat milk, and processed milk, or milk containing an equal or greater amount of non-fat milk solids, using lactic acid bacteria or yeast to form solid (hard), pasty (soft), or liquid (drinking) forms, or by freezing them, but is not limited to these. In the fermented milk of the present invention, the concentration range of non-fat milk solids is preferably 4.0% to 12.0%, more preferably 6.0% to 10.0%, and even more preferably 7.0% to 9.0%. In addition, the concentration of milk fat is preferably 0.2% to 4.0%, more preferably 0.3% to 3.0%, and even more preferably 0.4% to 2.0%.

[0044] A typical example of fermented milk is yogurt. In this invention, "yogurt" includes, for example, plain yogurt, hard yogurt (set-type yogurt), soft yogurt, and drinkable yogurt. From the viewpoint of ease of consumption when appetite is poor, drinkable yogurt is particularly preferred in this invention.

[0045] In the fermented milk of the present invention, the portion suitable for a single consumption is packaged in one piece, making it convenient and easy to consume, which is preferable from a usability perspective. The suitable portion for a single consumption varies from person to person depending on the degree of loss of appetite to be improved, but in the case of fermented milk with a nonfat milk solids content of 8.0%, for example, it is preferably 50 mL to 200 mL per serving, more preferably 80 mL to 150 mL, and even more preferably 100 mL to 120 mL. Alternatively, it is preferably 50 g to 200 g per serving, more preferably 80 g to 150 g, and even more preferably 100 g to 120 g.

[0046] In this invention, "the form of a single package" includes any form, including, for example, a container with a lid, a bottle with a lid, an individual bag, a pouch, a tube, and other general packaging forms. In this invention, the purpose of the product can be clearly defined by recording the product's uses, efficacy, method of consumption, etc., on a single package or a package containing multiple single packages, and / or by using additional informational packaging and / or by separately publishing informational materials such as brochures.

[0047] In this invention, for humans, the effective daily intake of the active ingredient, lactic acid bacteria, is preferably 2 × 10⁶. 7 ~5×10 10 One, or better yet, 5 x 10 7 ~5×10 10 One, even better is 1×10 8 ~5×10 10 One, and even better, 5 x 10 8 ~5×10 10 One, even better is 5×10 8 ~2×10 10 indivual.

[0048] The fermented milk of the present invention can achieve sufficient effects through a single intake. From the viewpoint of promoting the secretion of ghrelin, it is preferable to consume it continuously for a period of 4 weeks or more, 6 weeks or more, 8 weeks or more, 10 weeks or more, or 12 weeks or more; more preferably, for a period of 24 weeks or more; and even more preferably, for a period of 36 weeks or more. Furthermore, the fermented milk of the present invention is widely known and safe to consume; therefore, there is no particular upper limit to the intake period, and it can be continued indefinitely. However, if an upper limit must be set, it could be, for example, less than 120 weeks, less than 100 weeks, less than 80 weeks, or less than 60 weeks.

[0049] Furthermore, in this invention, commercially available products containing the lactic acid bacteria described in the embodiments of this invention can be conveniently used. For example, in the case of Lactobacillus gasseri OLL2716 (FERM BP-6999), "Meiji Probiotic Yogurt LG21" sold by Meiji Co., Ltd. can be conveniently used. This commercially available product can be consumed directly or further processed. When other ingestible ingredients are consumed together with the lactic acid bacteria described in the embodiments of this invention, there are no restrictions on the other ingestible ingredients, and milk-based ingredients are preferred. Milk-based ingredients refer to compositions containing milk itself or milk components obtained from processing milk, including all components containing milk components such as raw milk (cow's milk, etc.), reconstituted milk (milk powder, cream, butter), fermented milk (yogurt, cheese), and milk products (whey protein, casein, lactose, whey minerals, whey permeate), and their source and form are not particularly limited.

[0050] Furthermore, *Lactobacillus gasseri* OLL2716 is known to be a lactic acid bacterium with a high bactericidal ability against *Helicobacter pylori* in the human stomach (e.g., Japanese Patent No. 4509250). The ghrelin secretion-promoting effect of this invention can be obtained independently of *Helicobacter pylori* eradication. Therefore, this invention can be used in both *Helicobacter pylori*-positive and *Helicobacter pylori*-negative individuals. Example

[0051] The present invention will now be described in more detail based on embodiments. However, these embodiments do not limit the scope of the invention.

[0052] [Example 1]

[0053] Twelve male ddy mice (SLC Corporation, Japan, 4 weeks old) were divided into a control group and an LG21 yogurt group after a 1-week acclimatization period. The control group was fed AIN-93M diet (Coria Corporation, Japan, composition: corn starch 46.5692%, casein 14.0%, pregelatinized corn starch 15.5%, refined white sugar 10.0%, refined soybean oil 4.0%, cellulose powder 5.0%, mineral mixture (AIN-93M-MX) 3.5%, vitamin mixture (AIN-93VX) 1.0%, L-cysteine ​​0.18%, tartrate choline 0.25%, tert-butylhydroquinone 0.0008%) for 4 weeks. For the LG21 yogurt group, the diet was fed with AIN-93M feed containing LG21 yogurt (Meiji Probiotic Yogurt LG21 sold by Meiji Co., Ltd., containing Lactobacillus gasseri OLL2716 (FERM BP-6999)) at a dose of 1.11 g / day for 4 weeks. After 4 weeks, the animals were necropsyed and blood and stomach tissue were collected.

[0054] After EDTA treatment, 1 / 10 volume of hydrochloric acid (1 mol / L) was added to the blood, and the concentration of active ghrelin in the blood was measured using an active ghrelin ELISA kit (LSI Medience Corporation). In addition, cDNA was prepared from collected gastric tissue using a PrimeScript RT kit (Takara Bio Inc.), and the expression level of the ghrelin gene in the gastric tissue was measured using SYBER Premix Ex TaqII (Takara Bio Inc.) and Thermal cycler Dice (Takara Bio Inc.).

[0055] The results of the measurement of active ghrelin concentration in the blood are shown in Figure 1 The expression level of the ghrelin gene in gastric tissue was shown in Figure 2 .according to Figure 1 The serum ghrelin concentration in the LG21 yogurt group was significantly higher than that in the control group. Furthermore, according to... Figure 2 The expression level of the ghrelin gene in gastric tissue was significantly increased compared with the control group. Therefore, this suggests that LG yogurt containing Lactobacillus gasseri OLL2716 (FERM BP-6999) has a ghrelin-promoting effect.

[0056] [Example 2]

[0057] Fifteen male, 10-week-old ICR mice (SLC Corporation, Japan) were divided into three groups after a one-week acclimatization period. After a 5-hour fast, one group served as the control group and was given 500 μL of physiological saline. Gastric tissue was collected via necropsy. The other two groups were given 500 μL of saline containing 2 × 10⁻⁶ cells / mL of saline. 10 Live Lactobacillus gasseri OLL2716 (FERM BP-6999) in physiological saline was administered, and gastric tissue was collected by necropsy 1 and 2 hours later. The expression level of the ghrelin gene in the gastric tissue was measured using the same method as in Example 1.

[0058] The results are shown in Figure 3 Two hours after administration of live Lactobacillus gasseri OLL2716 (FERM BP-6999), the expression level of the ghrelin gene in gastric tissue was significantly increased compared with the control group. Therefore, this suggests that live Lactobacillus gasseri OLL2716 (FERM BP-6999) has a ghrelin-promoting effect.

[0059] Industrial applications

[0060] By employing this invention, the ghrelin secretion-promoting effect can be achieved through the ingestion of lactic acid bacteria themselves or fermented milk prepared using them, significantly improving loss of appetite. In particular, fermented milk is a widely known and readily available food, easily and safely consumed, thus providing substantial assistance in improving loss of appetite.

Claims

1. The use of fermented milk containing or prepared using *Lactobacillus gasseri* OLL2716 FERM BP-6999 in the preparation of a ghrelin secretion promoter for regulating gastrointestinal motility; wherein, The Lactobacillus gasseri OLL2716 FERM BP-6999 is the only effective ingredient for promoting ghrelin secretion.

2. The use as described in claim 1, characterized in that, The ghrelin secretion promoter contains 2×10 7 ~5×10 10 Lactobacillus gasseri OLL2716 FERM BP-6999.

3. The use as described in claim 1, characterized in that, The ghrelin secretion promoter is packaged in units of 50mL to 200mL.

4. The use as described in claim 1, characterized in that, The ghrelin secretion promoter is used for individuals who are Helicobacter pylori negative.

5. The use as described in claim 1, characterized in that, The fermented milk mentioned is yogurt.

6. Use of Lactobacillus gasseri OLL2716 FERM BP-6999 in the preparation of a ghrelin secretion-promoting composition for regulating gastrointestinal motility; in, The Lactobacillus gasseri OLL2716 FERM BP-6999 is the only effective ingredient for promoting ghrelin secretion; and the ghrelin secretion composition is yogurt. Furthermore, the described uses also include one or more features selected from the following group: a. The ghrelin-secreting composition contains 2×10 7 ~5×10 10 Lactobacillus gasseri OLL2716 FERM BP-6999; b. The ghrelin secretion composition is packaged in units of 50 mL to 200 mL; and d. The ghrelin secretion composition is used for Helicobacter pylori-negative individuals.

7. The use of *Lactobacillus gasseri* OLL2716 FERM BP-6999 in the preparation of fermented milk for regulating gastrointestinal motility by promoting ghrelin secretion; wherein, The Lactobacillus gasseri OLL2716 FERM BP-6999 is the only effective ingredient for promoting ghrelin secretion.

8. The use as described in claim 7, characterized in that, It also includes one or more features selected from the following group: a. The fermented milk contains 2×10 7 ~5×10 10 Lactobacillus gasseri OLL2716 FERM BP-6999; b. The fermented milk is packaged in units of 50mL to 200mL; d. The fermented milk is intended for individuals who are Helicobacter pylori negative; and e. The fermented milk is yogurt.

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