Composition for treating menstrual-associated symptoms

Abstract

To provide means for mitigating symptoms associated with menstruation using food, and also to provide a new use of Lactobacillus paragasseri.SOLUTION: Provided is a composition for treating menstrual-associated symptoms, comprising lactic acid bacteria classified as Lactobacillus paragasseri. This composition is especially effective for either physical discomfort symptoms before menstruation or mental discomfort symptoms before menstruation.SELECTED DRAWING: Figure 3

Description

[Technical field]

[0001] The present invention relates to compositions for the treatment of menstrual symptoms. [Background technology]

[0002] The Act on Promotion of Women's Participation and Advancement in the Workplace was enacted in 2015, and women's participation in society has progressed, but menstruation remains one of the major health issues for women. Symptoms such as irritability, lower abdominal pain, anger, headaches, breast pain, and lower abdominal pain during menstruation can develop into various problems such as poor quality of sleep, problems at work, and adverse effects on childcare, if the symptoms are severe, and can have a negative impact on physical, mental, and social aspects. In a survey report by Tanaka et al. (Non-Patent Document 1) targeting approximately 20,000 menstruating Japanese women aged 15 to 49, it was reported that 74% of women suffer from menstrual symptoms, and the estimated economic burden is approximately 682.8 billion yen per year. As such, symptoms associated with menstruation are a major problem that interferes with women's work and studies, but on the other hand, only 20% of women have ever visited a doctor, and many deal with the problem by using over-the-counter painkillers (Non-Patent Document 1).

[0003] Several effective ingredients have been investigated for treating menstrual symptoms. For example, Patent Document 1 provides a composition for treating menstrual symptoms, which contains α-lactalbumin.

[0004] On the other hand, Lactobacillus gasseri / paragasseri has been known as a lactic acid bacteria species that is normally present in the intestines of healthy people. So far, Lactobacillus paragasseri has been investigated as an active ingredient of a food and drink composition for preventing and / or improving endometriosis (Patent Document 2) and as an active ingredient of a stress-reducing agent (Patent Document 3).

[0005] Furthermore, agents for relieving premenstrual physical and / or mental discomfort (Patent Document 4, Non-Patent Document 2) and agents for improving female menopausal symptoms (Patent Document 5) containing a lactic acid bacteria strain, Lactobacillus gasseri CP2305 strain (accession number: FERM BP-11331), a processed product of the lactic acid bacteria strain, or an extract thereof, have been investigated. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] International Publication WO2019 / 235451 [Patent Document 2] International Publication WO2012 / 073924 [Patent Document 3] International Publication WO2014 / 132982 [Patent Document 4] International Publication WO2021 / 015104 [Patent Document 5] JP 2022-113373 A [Non-patent literature]

[0007] [Non-Patent Document 1] Tanaka E, Momoeda M, Osuga Y, et al. Burden of menstrual symptoms in Japanese women: results from a survey-based study. J Med Econ 2013: 16: 1255-1266. [Non-Patent Document 2] Nishida K, et al. Daily intake of Lactobacillus gasseri CP2305 ameliorates psychological premenstrual symptoms in young women : A randomized, double-blinded, placebo-controlled study. J Functional Foods 2021: 80: 104426 DISCLOSURE OF THEINVENTION [Problem to be solved by the invention]

[0008] If there were a food-based method that could alleviate menstrual symptoms, it would be beneficial for improving women's quality of life (QOL). [Means for solving the problem]

[0009] The present inventors have been actively studying Lactobacillus paragasseri. In the course of this investigation, they came up with the idea that this bacterial species may be effective against menstrual symptoms, and as a result, they have continued their research and completed the present invention. The present invention provides the following:

[0010] [1] A composition for treating menstrual symptoms, comprising lactic acid bacteria belonging to Lactobacillus paragasseri. [2] The composition described in 1, wherein the menstrual symptoms are either premenstrual physical discomfort or premenstrual mental discomfort. [3] The composition described in 1 or 2, wherein the menstrual symptoms are premenstrual mental discomfort symptoms. [4] A composition described in any one of 1 to 3, intended for ingestion by a subject not suffering from menopausal symptoms or a subject in adolescence or sexual maturity. [5] The composition described in any one of 1 to 4, wherein the menstrual symptom is a mental discomfort symptom, and the mental discomfort symptom is one or more selected from irritability, depression (low mood), intolerance, irritability, confusion, emotional instability, dissatisfaction, hopelessness, decreased energy, fatigue, and decreased motivation. [6] A composition for controlling premenstrual mood elevation, comprising Lactobacillus paragasseri. [7] A composition for controlling premenstrual irritability, comprising Lactobacillus paragasseri. [8] A composition described in any one of 1 to 7, containing Lactobacillus paragasseri as a heated bacterial cell. [9] A composition described in any one of 1 to 8, containing only Lactobacillus paragasseri cells as an active ingredient.

[10] The composition described in any one of 1 to 9, wherein Lactobacillus paragasseri is Lactobacillus paragasseri OLL2809 deposited under accession number NITE BP-72, or a taxonomically equivalent strain thereto.

[11] Lactobacillus paragasseri cells were administered at a daily dose of 1 × 10 8 ~1×10 12 11. The composition according to any one of 1 to 10, which is intended for individual ingestion.

[12] A method (excluding medical procedures) for treating menstrual symptoms, comprising a step of having a subject ingest a composition containing lactic acid bacteria belonging to Lactobacillus paragasseri.

[13] Lactobacillus paragasseri cells were administered at a daily dose of 1 × 10 8 ~1×10 12 13. The method according to claim 12, wherein the individual is ingested.

[0011] [1] A composition for treating menstrual symptoms, comprising lactic acid bacteria belonging to Lactobacillus paragasseri. [2] The composition described in 1, wherein the menstrual symptoms are either premenstrual physical discomfort or premenstrual mental discomfort. [3] The composition described in 1 or 2, wherein the menstrual symptoms are premenstrual mental discomfort symptoms. [4] A composition described in any one of 1 to 3, intended for ingestion by a subject not suffering from menopausal symptoms or a subject in adolescence or sexual maturity. [5] The composition described in any one of 1 to 4, wherein the menstrual symptom is a mental discomfort symptom, and the mental discomfort symptom is one or more selected from irritability, depression (low mood), intolerance, irritability, confusion, emotional instability, dissatisfaction, hopelessness, decreased energy, fatigue, and decreased motivation. [6] A composition for controlling premenstrual mood elevation, comprising Lactobacillus paragasseri. [7] A composition containing Lactobacillus paragasseri for controlling premenstrual irritability. [8] A composition described in any one of 1 to 7, containing Lactobacillus paragasseri as a heated bacterial cell. [9] A composition described in any one of 1 to 8, containing only Lactobacillus paragasseri cells as an active ingredient.

[10] The composition described in any one of 1 to 9, wherein Lactobacillus paragasseri is Lactobacillus paragasseri OLL2809 deposited under accession number NITE BP-72, or a taxonomically equivalent strain thereto.

[11] Lactobacillus paragasseri cells were administered at a daily dose of 1 × 10 8 ~1×10 12 11. A composition according to any one of claims 1 to 10, for individual ingestion.

[0012]

[21] Lac, for use in a method for treating menstrual symptoms. A composition comprising lactic acid bacteria belonging to Lactobacillus paragasseri. Use of lactic acid bacteria belonging to Lactobacillus paragasseri in the manufacture of a composition for treating menstrual symptoms. A method or non-therapeutic method for treating menstrual symptoms, comprising the step of administering to a subject a composition comprising lactic acid bacteria belonging to Lactobacillus paragasseri. Use or non-therapeutic use of a composition comprising lactic acid bacteria belonging to Lactobacillus paragasseri for treating menstrual symptoms.

[22] The composition, use in manufacture, method or non-therapeutic method, or use or non-therapeutic use described in 21, wherein the menstrual symptoms are either premenstrual physical discomfort or premenstrual mental discomfort.

[23] The use, method or non-therapeutic method, or use or non-therapeutic use in manufacturing as described in 21 or 22, wherein the menstrual symptoms are premenstrual mental discomfort symptoms.

[24] A composition, use in manufacture, method or non-therapeutic method, or use or non-therapeutic use as described in any one of 1 to 3 for ingestion by a subject not suffering from menopausal symptoms or by a subject in adolescence or sexual maturity.

[25] The composition, use in manufacturing, method or non-therapeutic method, or use or non-therapeutic use described in any one of 21 to 24, wherein the menstrual symptoms are mental discomfort symptoms, and the mental discomfort symptoms are one or more selected from irritability, depression (low mood), intolerance, irritability, confusion, emotional instability, dissatisfaction, hopelessness, decreased energy, fatigue, and decreased motivation.

[26] A composition comprising lactic acid bacteria belonging to Lactobacillus paragasseri for use in a method for controlling premenstrual mood elevation. Use of lactic acid bacteria belonging to Lactobacillus paragasseri in the manufacture of a composition for controlling premenstrual mood elevation. A method or non-therapeutic method for controlling premenstrual mood elevation comprising the step of administering to a subject a composition comprising lactic acid bacteria belonging to Lactobacillus paragasseri. Use or non-therapeutic use of a composition comprising lactic acid bacteria belonging to Lactobacillus paragasseri for controlling premenstrual mood elevation.

[27] A composition comprising lactic acid bacteria belonging to Lactobacillus paragasseri for use in a method for controlling premenstrual irritability. Use of lactic acid bacteria belonging to Lactobacillus paragasseri in the manufacture of a composition for controlling premenstrual irritability. A method or non-therapeutic method for controlling premenstrual irritability comprising the step of administering to a subject a composition comprising lactic acid bacteria belonging to Lactobacillus paragasseri. Use or non-therapeutic use of a composition comprising lactic acid bacteria belonging to Lactobacillus paragasseri for controlling premenstrual irritability.

[28] A composition, use in production, method or non-therapeutic method, or use or non-therapeutic use according to any one of claims 21 to 27, comprising Lactobacillus paragasseri as heated cells.

[29] A composition, use in production, method or non-therapeutic method, or use or non-therapeutic use according to any one of claims 21 to 28, comprising only Lactobacillus paragasseri cells as an active ingredient.

[30] The composition, use in production, method or non-therapeutic method, or use or non-therapeutic use according to any one of claims 21 to 29, wherein Lactobacillus paragasseri is Lactobacillus paragasseri OLL2809 deposited under accession number NITE BP-72, or a taxonomically equivalent strain thereto.

[31] Lactobacillus paragasseri cells were administered at a daily dose of 1 × 10 8 ~1×10 12 31. The composition, use in manufacture, method or non-therapeutic method, or use or non-therapeutic use of any one of claims 21 to 30 for administration to a subject. Effect of the Invention

[0013] Menstrual symptoms can be treated with a composition containing lactic acid bacteria belonging to the species Lactobacillus paragasseri. [Brief description of the drawings]

[0014] [Figure 1] Exam Schedule [Diagram 2] Flowchart showing the flow of subjects through each stage [Diagram 3] Change over time in premenstrual MDQ "mood elevation" score adjusted for explanatory variables (group, analgesic use score, stress score, exercise habits). Error bars indicate 95% confidence intervals. [Figure 4] Change from baseline in premenstrual MDQ "elevated mood" symptom score during the third menstrual cycle (Mean ± SE) [Diagram 5] Changes in premenstrual VAS "irritability" scores over time, adjusted for explanatory variables (group, analgesic use score, stress score, and exercise habits). Error bars indicate 95% confidence intervals. [Figure 6] Lactobacillus paragasseri OLL2809 16s rRNA gene (SEQ ID NO:1) DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] The present invention relates to the use of lactic acid bacteria belonging to Lactobacillus paragasseri for treating menstrual symptoms, and a composition therefor. Note that, although the present invention, embodiments, and examples may be described using a composition containing a specific lactic acid bacteria as an example, the description also applies to the use of the lactic acid bacteria, the method of use, etc.

[0016] [Active ingredients] (Lactobacillus paragasseri) The composition of the present embodiment uses lactic acid bacteria belonging to Lactobacillus paragasseri as an active ingredient. The lactic acid bacteria may be any of the specific strains. In the present invention, "any" is used to mean "at least one" unless otherwise specified. "Used as an active ingredient" refers to being used in an effective amount to exert the intended function, being used with the labeling specified as an ingredient contributing to the intended purpose, etc. In relation to the present invention, when describing bacteria belonging to the genus Lactobacillus, unless otherwise specified, the classification follows the reclassification by Zheng J, Wittouck S, Salvetti E, Franz CMAP, Harris HMB, Mattarelli P, O'Toole PW, Pot B, Vandamme P, Walter J, Watanabe K, Wuyts S, Felis GE, Ganzle MG, Lebeer S.: A taxonomic note on the genus Lactobacillus: Description of 23 novel genera, emended description of the genus Lactobacillus Beijerinck 1901, and union of Lactobacillaceae and Leuconostocaceae. Int J Syst Evol Microbiol. 2020 Apr; 70(4): 2782-2858.

[0017] Lactobacillus paragasseri is a gram-positive bacterium that performs homolactic fermentation and is classified as JCM5343. T The type strain is Lactobacillus paragasseri. Lactobacillus paragasseri can be distinguished from Lactobacillus gasseri by the average nucleotide identity (ANI) or DNA-DNA hybridization. An example of a lactic acid bacteria strain belonging to Lactobacillus paragasseri is JCM 5343. T , Lactobacillus paragasseri OLL2809 (NITE BP-72), FERM BP-6999, and NITE BP-224.

[0018] Strains with numbers beginning with JCM can be purchased from the Microbial Materials Development Laboratory, BioResource Research Center, RIKEN (3-1-1 Takanodai, Tsukuba, Ibaraki, Japan 305-0074). Strains with numbers beginning with NITE can be obtained from the National Institute of Technology and Evaluation Patent Microorganism Depositary (Room 122, 2-5-8 Kazusa Kamatari, Kisarazu, Chiba, Japan). Strains with numbers beginning with FERM can be obtained from the National Institute of Technology and Evaluation Patent Organism Depositary (Room 120, 2-5-8 Kazusa Kamatari, Kisarazu, Chiba, Japan).

[0019] In a preferred embodiment, the composition contains Lactobacillus paragasseri OLL2809 or a mycologically equivalent strain thereof as an active ingredient. As described below, Lactobacillus paragasseri OLL2809 is highly effective in treating menstrual symptoms, particularly unpleasant premenstrual symptoms.

[0020] (Lactobacillus paragasseri OLL2809) Lactobacillus paragasseri OLL2809 has been internationally deposited under the Budapest Treaty under accession number NITE BP-72. Depositor: Meiji Co., Ltd. Depository institution: National Institute of Technology and Evaluation, Patent Microorganisms Depository Center Accession number: NITE BP-72 Original deposit date: February 1, 2005 Date of transfer under the Budapest Treaty: January 18, 2006

[0021] The sequence of the 16S rRNA gene of Lactobacillus paragasseri OLL2809 (total length 1580 bases) is shown in FIG. 6 and SEQ ID NO: 1 in the sequence listing.

[0022] The sequence identity between the sequence of SEQ ID NO: 1 derived from Lactobacillus paragasseri and the sequence of the 16S rRNA gene of Lactobacillus gasseri (Sequence ID: NR_075051.2) is 99.24% (1568 / 1580 matches). In one embodiment, the Lactobacillus paragasseri contained in the composition has a 16S rRNA gene sequence that has a sequence identity of 99.25% or more with the sequence of SEQ ID NO:1, preferably 99.3% or more, more preferably 99.35% or more, even more preferably 99.4% or more, even more preferably 99.45% or more, even more preferably 99.5% or more, even more preferably 99.55% or more, even more preferably 99.6% or more, even more preferably 99.65% or more, even more preferably 99.7% or more, even more preferably 99.75% or more, even more preferably 99.8% or more, even more preferably 99.85% or more, even more preferably 99.9% or more, even more preferably 99.95% or more, and even more preferably 100%.

[0023] The mycological properties of Lactobacillus paragasseri OLL2809 are shown below. Colony properties on medium (Lactobacilli MRS Agar, DIFCO): 2-3 mm in diameter, pale yellow, round, flat, full-edged, smooth, opaque, sticky Bacterial form: Bacillus Gram stain: positive Lactic acid fermentation type: homolactic fermentation Aerobic growth: + Growth temperature: 15℃ -, 45℃ + Sugar fermentation: arabinose -, xylose -, rhamnose -, ribose -, glucose +, mannose +, fructose +, galactose +, sucrose +, cellobiose +, lactose +, trehalose +, melibiose -, raffinose -, melitzose -, mannitol -, sorbitol -

[0024] A strain taxonomically equivalent to a certain strain (hereinafter referred to as strain S) refers to, for example, any of the following: The entire sequence of its 16S rRNA gene (hereinafter sometimes referred to as "16S" or "16S rRNA gene") or a characteristic part thereof (such as the V1 region, the V2 region, or all or part of the V1 region and the V2 region, or a part including the V1 region and the V2 region, etc.) has a sequence identity of 97% or more, more preferably 98% or more, even more preferably 98.5% or more, even more preferably 98.7% or more, even more preferably 99% or more, even more preferably 99.25% or more, even more preferably 99.3% or more, even more preferably 99.35% or more, or a strain having a sequence identity of preferably 99.4% or more, more preferably 99.45% or more, even more preferably 99.5% or more, even more preferably 99.55% or more, even more preferably 99.6% or more, even more preferably 99.65% or more, even more preferably 99.7% or more, even more preferably 99.75% or more, even more preferably 99.8% or more, even more preferably 99.85% or more, even more preferably 99.9% or more, even more preferably 99.95% or more, even more preferably 100%; A strain with the same mycological properties as strain S. A mutant or derivative of strain S, which has the effect of treating menstrual symptoms.

[0025] In the present invention, unless otherwise specified, sequence identity means the ratio of the number of identical bases shared between two sequences when the two sequences are optimally aligned. Analysis of the identity of base sequences can be performed using algorithms or programs (e.g., BLASTN, BLASTP, BLASTX, ClustalW) well known to those skilled in the art. When using a program, the parameters can be appropriately set by those skilled in the art, or the default parameters of each program may be used. Specific techniques for these analysis methods are also well known to those skilled in the art. For calculating identity, commercially available genetic information processing software may be used.

[0026] Regarding criteria for determining species similarity based on 16S rRNA gene sequences, those skilled in the art can refer to Stackebrandt E, Ebers J. Taxonomic parameters revisited: tarnished gold standards. Microbiol Today 2006;33:152-155.

[0027] (Other active ingredients) In one embodiment, the active ingredient in the composition is only the bacterial cells of lactic acid bacteria belonging to Lactobacillus paragasseri, for example, only the bacterial cells of Lactobacillus paragasseri OLL2809 deposited under the accession number NITE BP-72, or a taxonomically equivalent strain. That is, in one embodiment, the composition does not contain other components known to be effective for treating menstrual symptoms, such as α-lactalbumin, other than the bacterial cells of lactic acid bacteria belonging to Lactobacillus paragasseri, in an amount that is clinically effective for treating menstrual symptoms. When the composition comprises alpha-lactalbumin, an amount less than the amount which is clinically effective for treating menstrual symptoms means an amount less than 100 mg per serving, for example 50 mg or less, 25 mg or less, 20 mg or less, 10 mg or less, 5 mg or less, 3 mg or less, 2 mg or less, 1 mg or less, 0.5 mg or less, 0.3 mg or less, 0.2 mg or less, 0.1 mg or less.

[0028] (Manufacturing method) The lactic acid bacteria belonging to Lactobacillus paragasseri contained in the composition can be produced by culturing. The culturing conditions are not particularly limited as long as the desired effect is achieved.

[0029] In the composition, the lactic acid bacteria belonging to Lactobacillus paragasseri may be contained in any state as long as the desired effect can be exhibited. For example, the lactic acid bacteria may be the lactic acid bacteria itself or a culture of the lactic acid bacteria (consisting of the lactic acid bacteria and culture supernatant).

[0030] The lactic acid bacteria culture can be obtained according to a conventional method for preparing a lactic acid bacteria culture. Various media can be used for the culture, such as a milk medium or a medium containing a milk component, a semi-synthetic medium not containing the milk component, or a synthetic medium. The lactic acid bacteria culture can be obtained by inoculating the lactic acid bacteria belonging to Lactobacillus paragasseri into the sterilized medium and culturing at about 20°C to 45°C for 5 hours to several days. The temperature and period of the culture can be appropriately adjusted depending on the lactic acid bacteria strain belonging to Lactobacillus paragasseri used and the desired number of bacteria. The culture may be used as it is, or may be used after processing such as centrifugation, membrane concentration, drying, freeze-drying, and washing.

[0031] The bacterial cells may be in a living state (viable bacterial cells) or in a dead state (dead bacterial cells) so long as they are capable of exerting the intended effect.

[0032] Killed bacteria can be obtained by sterilizing lactic acid bacteria. The sterilization is not particularly limited as long as it can exert the desired effect, and can be performed by heating, a germicidal lamp (UV), ozone, a drug, high osmotic pressure, etc. The killed bacteria are preferably heat-killed bacteria obtained by heat-treating live bacteria. The heat treatment for obtaining heat-killed bacteria is not particularly limited as long as it exerts the desired effect, and is performed at a temperature and time sufficient to kill the lactic acid bacteria used. Although such conditions vary depending on the lactic acid bacteria used, they are, for example, 55°C or higher, preferably 60°C or higher, more preferably 65°C or higher, and even more preferably 70°C or higher, and may be 80°C or higher, or may be 90°C or higher. The upper limit of the heat treatment temperature can be appropriately set, for example, 121°C or lower, 100°C or lower, 90°C or lower, or 80°C or lower. Depending on the heat treatment temperature, the heat treatment time can be 1 minute or more, 3 minutes or more, 10 minutes or more, 15 minutes or more, 30 minutes or more, or 45 minutes or more. The upper limit of the heat treatment time can be, for example, 120 minutes or less, may be 100 minutes or less, may be 90 minutes or less, or may be 80 minutes or less.

[0033] The lactic acid bacteria to be contained in the composition, regardless of whether they are live or killed bacteria, can be prepared in the form of a washed bacteria product, concentrate, dried product, suspension, paste, gel, etc.

[0034] [Application] (Functions, effects, etc.) The composition of this embodiment can be used for the treatment of menstrual symptoms. The treatment includes suppressing, inhibiting, or reducing the onset and appearance (onset) of the target disease or condition, reducing the risk of onset and onset, treating the onset and onset of the target disease or condition, and suppressing, inhibiting, or delaying the progression of the target disease or condition. The treatment includes medical procedures performed by doctors and nurses and midwives under the instructions of doctors, and non-therapeutic procedures performed by persons other than doctors, such as pharmacists, nutritionists (including dietitians and sports nutritionists), public health nurses, midwives, nurses, clinical laboratory technicians, sports instructors, pharmaceutical manufacturers, pharmaceutical distributors, food manufacturers, food distributors, etc. The treatment further includes recommendations for the intake of specific foods and nutritional guidance (including nutritional guidance necessary for the medical treatment of injured and sick people, and nutritional guidance for maintaining and promoting health).

[0035] Menstrual symptoms are a general term for unpleasant symptoms that accompany menstruation, and include unpleasant symptoms before and during menstruation. Menstrual symptoms include premenstrual syndrome (PMS) and premenstrual dysphoric disorder (PMDD). PMS is defined as "mental or physical symptoms that last 3 to 10 days before menstruation and that subside or disappear with the onset of menstruation." When the mental symptoms are predominant and strong among PMS, it is diagnosed as premenstrual dysphoric disorder (PMDD). PMDD is mainly characterized by four symptoms: depressed mood, anxiety / tension, emotional instability, and anger / irritability. Characteristic symptoms such as changes in eating behavior and sleep disorders appear before menstruation, which interfere with social activities and human relationships, but the cause and pathology are not yet fully understood. It improves or disappears with the onset of menstruation.

[0036] In the present invention, the term "premenstruation" refers to the period from 7 days before the expected onset of menstruation to the day before the start of menstruation, unless otherwise specified. Premenstruation can also be referred to as the period after ovulation and before menstruation when body temperature rises (high temperature period). Body temperature varies from person to person, but during the high temperature period, body temperature usually rises by about 0.3 to 0.5°C.

[0037] Menstrual symptoms are distinguishable from female menopausal symptoms. Menstrual symptoms appear temporarily before or during menstruation in accordance with the sexual cycle, and the symptoms weaken or disappear as the menstrual period progresses, whereas female menopausal symptoms do not show the same periodicity as menstrual symptoms. In addition, as described below, subjects with menstrual symptoms and subjects with female menopausal symptoms can also be distinguished as subjects.

[0038] The evaluation of whether an ingredient is useful for treating menstrual symptoms can be done by the Menstual Distress Questionaire (MDQ) (Moos RH. The development of a menstrual distress questionnaire. Psychosom Med 1968: 30: 853-867.) In addition to the MDQ, it can also be done by visual analog scales (VAS) and the Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36).

[0039] In detail, the MDQ is a 46-item self-report questionnaire for use in the assessment and treatment of premenstrual and menstrual symptoms. The 46 items are categorized into eight subfactors: (1) pain, (2) fluid retention, (3) autonomic nervous system changes, (4) negative affect, (5) poor concentration, (6) behavioral changes, (7) mood elevation, and (8) control. Of these subfactors, only (7) mood elevation captures positive changes. The SF-36 can also be used to evaluate health-related QOL. More specifically, the SF-36 can be used to score eight subscales: (1) physical functioning, (2) role functioning (physical), (3) bodily pain, (4) general health, (5) vitality, (6) social functioning, (7) role functioning (mental), and (8) mental health. These subscales can then be divided into physical health, mental health, and role / social health and scored accordingly. A higher score indicates a better health condition. In addition, the VAS can be used to rate the intensity of five symptoms: (1) lower abdominal pain, (2) rough skin, (3) swelling, (4) irritability, and (5) depression, once a day from seven days before the expected start of menstruation through the fourth day of menstruation.

[0040] More specifically, subjects who are aware of menstrual symptoms are asked to take the test ingredient once to several times a day for several sexual cycles, for example, three sexual cycles, and the evaluation items are investigated before menstruation (for example, from 7 days before the day when menstruation is expected to occur to the day before the start of menstruation) and during menstruation (for example, from the first day of menstruation to the fourth day of menstruation) at the time of menstruation such as before the intake and the first and second intakes after the intake. As a result, if improvement in at least one symptom is observed in the intake group, the ingredient can be evaluated as effective. The evaluation may be performed by comparing with a placebo intake group, or may be performed by comparing with before intake.

[0041] The composition of the present embodiment is useful for treating either premenstrual physical discomfort or premenstrual mental discomfort, among menstrual symptoms, and is particularly useful for treating premenstrual mental discomfort.

[0042] Specific examples of mental discomfort symptoms include irritability, depression (low mood), intolerance, irritability, confusion, emotional instability, dissatisfaction, hopelessness, loss of vitality, fatigue, and loss of motivation. Irritability can be evaluated, for example, by a VAS. Depression (low mood) can be evaluated, for example, by the MDQ as a change in the score for "elevated mood." Intolerance and irritability can be evaluated, for example, by the MDQ as a change in the score for "feeling lovable toward people." Confusion and emotional instability can be evaluated, for example, by the MDQ as a change in the score for "feeling things are organized." Dissatisfaction can be evaluated, for example, by the MDQ as a change in the score for "feeling happy." Hopelessness, loss of vitality, fatigue, and loss of motivation can be evaluated, for example, by the MDQ as a change in the score for "feeling energetic and active."

[0043] Physical discomfort may include lower abdominal pain, dry skin, and swelling.

[0044] In one embodiment, the composition is used to regulate premenstrual mood elevation. In the context of the present invention, regulation includes upregulating (increasing, enhancing) and downregulating (decreasing, suppressing). In a particularly preferred embodiment, the composition is used to enhance premenstrual mood elevation.

[0045] In another embodiment, the composition is used to control premenstrual irritability. In a particularly preferred embodiment, the composition is used to suppress premenstrual irritability.

[0046] The composition of the present embodiment is expected to be useful for treating diseases whose symptoms are known to worsen with the menstrual cycle, such as dysmenorrhea (functional and organic), depression, anxiety disorder (panic disorder), allergies, and the transition period to menopause.

[0047] In one aspect, the composition of the present embodiment can be used for non-disease treatment purposes, for menstrual symptoms that do not correspond to a disease. Menstrual symptoms that do not correspond to a disease refer to conditions that do not affect daily life and do not require medical treatment.

[0048] (subject) The composition of the present embodiment is suitable for administration to subjects for whom it is desirable or necessary to treat menstrual symptoms: subjects for whom it is desirable or necessary to treat either premenstrual physical discomfort symptoms or premenstrual mental discomfort symptoms: subjects for whom it is desirable or necessary to treat one or more selected from irritability, depression (low mood), intolerance, irritability, confusion, emotional instability, dissatisfaction, hopelessness, low energy, fatigue, and low motivation; subjects not suffering from menopausal symptoms; subjects in adolescence or sexual maturity; subjects for whom it is desirable or necessary to control premenstrual mood elevation; subjects for whom it is desirable or necessary to control premenstrual irritability. Desirable subjects include those who are aware of such symptoms themselves.

[0049] The subject to which the composition of the present embodiment is administered can be distinguished from the subject having female menopausal symptoms. Women usually go through four life stages (puberty, sexual maturity, menopause, old age) with age, and although there are individual differences, they reach menopause at around 50 years old, and the 10 years before and after this menopause (generally around 45 to 55 years old) are called "menopause". However, it is possible to distinguish between subjects in menopause and those not in menopause (subjects in adolescence or sexual maturity), and the composition of the present embodiment can be administered to subjects not suffering from menopausal symptoms.

[0050] The age of the subject is not particularly limited, but may be, for example, 18 years or older, 25 years or older, or 45 years or younger, or 40 years or younger.

[0051] The composition of the present embodiment is particularly suitable for administration to women who experience physical or mental discomfort, especially mental discomfort, before menstruation, but not severe enough to require a visit to the hospital, or not severe enough to affect their daily life, or who do not wish to seek medical treatment from a doctor, and to healthy women aged 25 to 40 years, whose past six menstrual cycles have been within the range of 25 to 38 days, whose usual menstrual period lasts for 3 to 7 days, and who are aware of some menstrual symptoms before and during menstruation.

[0052] In the present invention, a healthy woman refers to a woman who does not fall under the following categories: a pregnant or lactating woman; a woman with a history of or current gynecological disease; a woman with a food allergy; a woman who suffers from diarrhea when ingesting dairy products; a woman who has the habit of taking analgesics preventively before the onset of menstrual pain; a woman with severe menstrual symptoms that cannot be controlled with over-the-counter analgesics; a woman whose neurosis discrimination chart based on the Cornell Medical Index (CMI) health questionnaire is in the IV region; a woman whose stress score based on the Simple Stress Checklist is 20 or more; and any other person who a doctor has determined to be inappropriate for administration of the composition of this embodiment.

[0053] [Composition] (Food composition, etc.) The composition of the present embodiment can be a food composition or a pharmaceutical composition. The food and pharmaceutical products include not only those for humans, but also those for animals other than humans, unless otherwise specified. The food includes general food, functional food, and nutritional composition, as well as therapeutic food (for the purpose of treatment. A doctor issues a meal prescription, and a nutritionist or the like prepares a menu based on the prescription.), dietary therapy food, ingredient-adjusted food, nursing care food, and food for medical support, unless otherwise specified. The food includes not only solids, but also liquids, such as beverages, drinks, liquid foods, and soups, unless otherwise specified. The functional food refers to food that can impart a specific functionality to a living body, and includes, for example, health functional foods including foods with specified health uses (including conditional FOSHU [specified health foods]), functional foods, foods with nutritional functions, foods with special uses, nutritional supplements, health supplements, supplements (for example, tablets, coated tablets, sugar-coated tablets, capsules, liquids, and other various dosage forms), and beauty foods (for example, diet foods). In addition, in the present invention, the term "functional food" includes health foods to which a health claim based on the food standards of Codex Alimentarius (the Joint FAO / WHO Food Standards Commission) is applied. The composition may also be a refined product of a specific sugar. Refined products include fractions, partially refined products, and crude refined products.

[0054] (Route of administration, etc.) The composition of the present embodiment may be administered orally, parenterally, for example, through a tube (gastrostomy, enterostomy), or nasally, but is preferably administered orally. In the present invention, administration is used not only to mean administering a pharmaceutical product to a subject, but also to mean having a subject ingest food or the like other than pharmaceutical products. "Administer" can be read as "intake," and "ingestion" can be read as "administer."

[0055] The composition can be administered to a subject repeatedly, and can be administered continuously for a long period of time. The period is not particularly limited, but in order to fully demonstrate the effect, it is preferable to administer the composition continuously for a relatively long period of time, for example, 3 days or more, 1 week or more, 2 weeks or more, 1 month or more, 3 months or more, 6 months or more, 1 year or more, or 1 sexual cycle or more, 2 sexual cycles or more, 3 sexual cycles or more, 4 sexual cycles or more, 5 sexual cycles or more, 6 sexual cycles or more.

[0056] The compositions may be administered routinely, proactively, such as when at high risk, or when the need arises. The compositions may be administered with a meal, before, after, or between meals, or at the onset of the disease or condition that it is desired to ameliorate with the composition.

[0057] (dose, content) The dosage of the composition of the present embodiment may be any amount that exerts the intended effect. The dosage may be appropriately determined taking into consideration various factors such as the age, weight, and symptoms of the subject.

[0058] The daily dose of the composition can be 5 mg or more, 10 mg or more, preferably 50 mg or more, more preferably 100 mg or more, in terms of the amount of active ingredient. The upper limit of the active ingredient per day, regardless of the lower limit, can be 1000 mg or less, 900 mg or less, 700 mg or less, 500 mg or less, 400 mg or less, 300 mg or less, or 200 mg or less. When a composition contains multiple active ingredients, the amount of the active ingredient refers to the total amount of the active ingredients contained. The administration may be once a day, or may be divided into multiple times a day, for example, 2 to 10 times.

[0059] The daily dose of the composition is 1×10 8 It can be more than 2 x 10 8 It can be 5 x 10 or more.8 It is preferable to have more than 1×10 9 It is preferable to have more than 2×10 9 More preferably, 5×10 9 More preferably, it is 1×10 9 It is more preferable that the upper limit of the daily dose of the active ingredient is 1×10 or more in terms of the amount of bacterial cells, regardless of the lower limit. 12 The number can be less than 5 x 10 11 The number can be less than 2 x 10 11 The number can be less than 5 x 10 10 The number can be less than 2 x 10 10 It may be less than one.

[0060] The content of the active ingredient in the composition can be appropriately determined according to the form of the composition. For example, when the composition is in a form to be eaten or consumed as is, such as fermented milk or beverage, the content of the active ingredient per 100 g of the composition can be 0.01% or more, 0.04% or more, preferably 0.1% or more, more preferably 0.2% or more, even more preferably 0.5% or more, preferably 1% or more, preferably 10% or more, and preferably 15% or more. The upper limit of the active ingredient per 100 g of the composition, regardless of the lower limit, can be 80% or less, 50% or less, 30% or less, or 20% or less. Alternatively, the content of the active ingredient per solid content of the composition can be 1% or more, more preferably 5% or more, more preferably 10% or more, and particularly preferably 15% or more. The upper limit of the active ingredient per solid content, whatever the lower limit, may be 80% or less, 50% or less, 40% or less, or 20% or less. In the present invention, % means mass % unless otherwise specified.

[0061] (Other ingredients, additives) In the present invention, the composition may contain other active ingredients or nutritional ingredients that are acceptable as foods or pharmaceuticals. Examples of such ingredients include lipids (e.g., milk fat, vegetable oils and fats, oils and fats containing medium-chain fatty acids), proteins (e.g., milk proteins, milk protein concentrates (MPC), whey protein concentrates (WPC), whey protein isolates (WPI), α-lactalbumin (α-La), β-lactoglobulin (β-Lg), heat-denatured whey proteins, and enzyme-treated whey proteins), amino acids (e.g., lysine, arginine, glycine, alanine, glutamic acid, leucine, isoleucine, valine), kojibiose, and sugars containing kojibiose as a constituent sugar. These include carbohydrates other than oligosaccharides (glucose, sucrose, fructose, maltose, trehalose, erythritol, maltitol, palatinose, xylitol, dextrin), electrolytes (e.g., sodium, potassium, calcium, magnesium), vitamins (e.g., vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin D, vitamin E, vitamin K, biotin, folic acid, pantothenic acid, and nicotinic acid), minerals (e.g., copper, zinc, iron, cobalt, manganese), antibiotics, and dietary fiber.

[0062] In the present invention, the composition may contain prebiotics. Prebiotics are indigestible food ingredients that selectively alter the growth and activity of specific bacteria in the large intestine, thereby beneficially affecting the host and improving the host's health. The composition may contain one or more prebiotics other than the active ingredient.

[0063] The prebiotics are not particularly limited as long as they do not interfere with the effects of the active ingredients contained in the composition. Examples of prebiotics other than the active ingredients include galactooligosaccharides, fructooligosaccharides (GF2, GF3, GF4), xylooligosaccharides, isomaltooligosaccharides, raffinose, lactulose, lactosucrose, soybean oligosaccharides, coffee oligosaccharides, dietary fiber, and gluconic acid.

[0064] The composition can also be used as a synbiotic, and may contain probiotics in addition to the active ingredient. Synbiotics are a combination of probiotics and prebiotics. Probiotics can be defined as microorganisms that have a beneficial effect on the host when introduced into the intestine of the host in a live state.

[0065] The probiotics are not particularly limited as long as they do not interfere with the effects of the active ingredients contained in the composition. Examples of probiotics include certain lactic acid bacteria.

[0066] The composition may further contain additives that are acceptable for use as food or medicine. Examples of such additives include inert carriers (solid or liquid carriers), excipients, surfactants, binders, disintegrants, lubricants, solubilizers, suspending agents, coating agents, colorants, preservatives, buffers, pH adjusters, emulsifiers, stabilizers, sweeteners, antioxidants, flavors, acidulants, and natural products. More specifically, these include water, other aqueous solvents, pharma- ceutically acceptable organic solvents, collagen, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymers, sodium alginate, water-soluble dextran, water-soluble dextrin, sodium carboxymethyl starch, pectin, xanthan gum, gum arabic, casein, gelatin, agar, glycerin, propylene glycol, polyethylene glycol, petrolatum, paraffin, stearyl alcohol, stearic acid, human serum albumin, mannitol, sorbitol, lactose, sucralose, stevia, aspartame, acesulfame potassium, citric acid, lactic acid, malic acid, tartaric acid, phosphoric acid, acetic acid, fruit juice, vegetable juice, and the like.

[0067] (Dosage form / shape) In one embodiment, the food composition may be prepared in any form, such as a solid, liquid, mixture, suspension, powder, granule, paste, jelly, gel, capsule, etc. The food composition according to the present embodiment may be in any form, such as a dairy product, a supplement (e.g., a tablet, a coated tablet, a sugar-coated tablet, an enteric coating, etc., a capsule (an enteric soft capsule, an enteric hard capsule, a large intestine delivery capsule, etc.), a confectionery, a beverage, a drink, a seasoning, a processed food, a side dish, a soup, etc. More specifically, the composition according to the present embodiment may be in any form, such as a liquid food, a semi-liquid food, a jelly, a gel, a powder, a formula, a liquid formula, a powder or liquid milk for pregnant women and lactating women, a fermented milk, a bar, a mousse, a chocolate, a biscuit, an ice cream, a fermented milk, a lactic acid bacteria drink, a dairy drink, a milk drink, a soft drink, a fruit juice drink, a tablet, a cheese, a bread, a biscuit, a cracker, a pizza crust, a food for the sick, a nutritional food, a frozen food, a processed food, etc. It may also be in the form of granules, powder, paste, thickened liquid, etc., for mixing with beverages or foods for administration. The granules and powders may be in the form of cubes or sticks (each packaged in a single serving). In the present invention, modified milk powder refers to raw milk, cow's milk, special cow's milk, raw buffalo milk, or food made from these ingredients, processed or used as the main ingredient, to which nutrients necessary for infants are added, and made into a powder, as defined in the "Ministerial Ordinance on the Compositional Standards, etc. of Milk and Dairy Products (hereinafter referred to as the "Ministerial Ordinance on Milk, etc."). In the present invention, modified liquid milk refers to raw milk, cow's milk, special cow's milk, raw buffalo milk, or food made from these ingredients, processed or used as the main ingredient, to which nutrients necessary for infants are added, and made into a liquid, as defined in the Ministerial Ordinance on Milk, etc.

[0068] In one aspect, the pharmaceutical composition can be in any dosage form suitable for oral administration, such as a solid formulation, such as a tablet, granule, powder, pill, or capsule; a liquid formulation, such as a solution, suspension, or syrup; a gel; or an aerosol.

[0069] (Manufacturing method, etc.) In one embodiment, in the production of the composition, the stage of blending the active ingredient can be appropriately selected. The stage of blending is not particularly limited as long as it does not significantly impair the properties of the active ingredient. For example, the active ingredient can be blended by mixing it with the raw material. Alternatively, the active ingredient can be added at the final stage of production to produce a composition containing the active ingredient.

[0070] (display) In one embodiment, the composition can be labeled with its intended use (application). The labeling can be made in accordance with the labeling laws and regulations of the country in which the present invention is implemented. In one embodiment, the function of the composition or active ingredient or the usage based on the function is labeled. Examples of the usage based on the function to be displayed are as described above in relation to the functions, actions, and effects. In addition, the composition of this embodiment can be labeled with the fact that it can be used as a prebiotic or as a synbiotic (a combination of probiotics and prebiotics).

[0071] In one embodiment, the composition is labeled with a recommendation for administration to a particular subject. Examples of subjects for which the labeling is provided are as described above for subjects.

[0072] Representation can be explicit or implicit. Examples of explicit representation are direct description on tangible objects such as the product itself, packaging, containers, labels, tags, etc., while examples of implicit representation (which can also be called implied) include advertising and promotional activities by place or means such as websites, storefronts, pamphlets, exhibitions, media seminars and other seminars, books, newspapers, magazines, television, radio, online video sharing platforms, SNS, mailings, emails, and audio.

[0073] In one embodiment, the recommendation to take the composition is displayed personally. Such a display can be made by using a document (whether written or electronic) addressed to the subject, a tablet terminal, a smartphone, a personal computer, a SNS of the subject, etc. Also, such a display can be made together with the display of the results of any test or analysis, such as a subject's symptom test, on the subject. EXAMPLES

[0074] [Manufacturing] The raw materials and mixing ratios of the test foods and control foods evaluated below are as shown in the table below. The powders uniformly mixed in the following ratios were compressed into tablets by conventional methods.

[0075] [Table 1]

[0076] [evaluation] 1. Method 1-1. Study design and implementation structure This study was commissioned to Sogo Medical Science Institute Co., Ltd. and was conducted in compliance with the ethical principles based on the Declaration of Helsinki and the ethical guidelines for medical research involving human subjects (notification by the Ministry of Education, Culture, Sports, Science and Technology and the Ministry of Health, Labor and Welfare, partially revised on February 28, 2017). This study was approved by the Meiji Clinical Trial Review Board (review number: Review-202) and the Ethics Committee of Fukuda Naika Clinic, Koseikai Medical Corporation (review number: IRB-20210619-3) and was conducted at Fukuda Naika Clinic, Koseikai Medical Corporation from May 2021 to September 2022. The study plan was registered in the UMIN Clinical Trial Registry (UMIN-CTR) (study ID: UMIN000044933) and was conducted with the consent of all subjects.

[0077] 1-2. Subject inclusion and exclusion criteria [Selection Criteria] Healthy adult women aged 25 to 40 at the time of obtaining informed consent -Those whose past six menstrual cycles were within the range of 25-38 days and whose usual menstrual period lasted 3-7 days -Those who experience any menstrual symptoms before or during menstruation -Those who have been fully informed of the purpose and contents of this study, are able to consent, have fully understood the study, and have volunteered of their own volition and agreed to participate in the study in writing

[0078] [Exclusion criteria] - Those who are pregnant, breastfeeding or possibly pregnant -Those with a history or current gynecological disease Those with food allergies -Those who experience diarrhea when consuming dairy products -Medicines (including herbal medicines and pills) that may affect this study from two months before consent acquisition to the day consent is acquired Those who regularly use supplements -Those who consume foods that may affect the test results (foods and beverages containing lactic acid bacteria such as yogurt) 4 days or more a week -Those who participated in clinical trials of other drugs or foods from one month before obtaining consent to the day of obtaining consent -Those who have a habit of taking analgesics as a preventative measure before the onset of menstrual pain. - Subjects whose postmenstrual MDQ score is 1 point or more higher than their premenstrual and menstrual MDQ scores in the Menstual Distress Questionaire (MDQ) (Non-Patent Document 4) at the first screening -Those whose symptoms are severe and cannot be controlled with over-the-counter pain medication Those who are in the IV region of the neuroticism scale according to the Cornell Medical Index (CMI) health questionnaire -Those with a stress score of 20 or more on the Simple Stress Checklist -Others who the investigator judges to be inappropriate for this study

[0079] During the study period, subjects were instructed to refrain from excessive dieting or overeating that deviated significantly from the normal range. They were also instructed to refrain from taking painkillers for preventive purposes. In order to help them distinguish between temporary menstrual symptoms in healthy women and chronic discomfort caused by gynecological diseases, they were instructed to immediately seek medical attention if premenstrual or menstrual discomfort interfered with daily life, or if the discomfort continued not only before or during menstruation but also after menstruation.

[0080] 1-3. Test food The OLL2809 heat-treated bacteria involved in this study were produced at Meiji Tokyo Lactic Acid Bacteria Factory, the freeze-dried powder was produced at Meiji Innovation Center, and tableting and packaging were carried out at API Ikeda Factory (Ibi District, Gifu Prefecture).

[0081] Test food: 100 mg of OLL2809 heat-treated bacteria per 2 tablets (1 × 10 10 Tablets containing Control food: A tablet candy in which the above active ingredients were replaced with freeze-dried powder of OLL2809 culture medium and dextrin.

[0082] 1-4. Test procedure For subjects who provided consent, blood and urine samples were taken, physical measurements were taken, and the Living Conditions Questionnaire, CMI Health Questionnaire (Non-Patent Document 5), Simple Stress Checklist (Katsura-Murakami version) (Non-Patent Document 6), MDQ, and interview were conducted as the first screening. From among the subjects who met the inclusion criteria and did not violate the exclusion criteria, 120 subjects with the highest MDQ total scores in the first screening were selected.

[0083] The following symptoms were evaluated using a visual analog scale (VAS): (1) lower abdominal pain, (2) rough skin, (3) swelling, (4) irritability, and (5) depression, from 7 days before the expected onset of menstruation in the -1st menstrual cycle to the 4th day of menstruation (Figure 1). On the first and 4th days of menstruation, the Medical Outcomes Study 36-Item Short-Form Health Survey v2 acute version (SF-36v2) (Non-Patent Document 7) and MDQ were administered. Subjects were asked to answer on the first day of menstruation about any changes in their physical and mental state that they had noticed during the period from 7 days before the start of menstruation to the day before, and on the 4th day of menstruation about any changes in their physical and mental state that they had noticed during the period from the first day of menstruation to the 4th day of menstruation (Figure 1). Among the subjects who met the inclusion criteria and did not violate the exclusion criteria, 80 subjects whose MDQ total score in the second screening was close to the median were selected and assigned to two groups.

[0084] The test food was administered once a day from the fifth day of menstruation in the 0th sexual cycle to the fourth day of menstruation in the 3rd sexual cycle. In the 2nd and 3rd sexual cycles, VAS, SF-36, and MDQ were administered in the same manner as in the -1st sexual cycle (Figure 1).

[0085] During the period from the start date of the secondary screening test to the end date of the test, subjects were asked to enter information in a daily diary about their intake of the test foods, subjective symptoms of changes in physical condition, treatments received, medicines used, and health foods used. Analgesic use scores and test food intake rates were calculated based on the records in the daily diary. Analgesic use scores were calculated as follows: "no use of analgesics" was scored as 0 points, "use once" as 1 point, "use twice" as 2 points, and "use three or more times" as 3 points. The value for the seven days from 7 days before menstruation to the day before menstruation was considered the "premenstrual" value, and the value for the four days from the first day of menstruation to the fourth day of menstruation was considered the "menstrual" value.

[0086] 1-5. Evaluation items The primary efficacy outcome measure in this study was the MDQ score, and the secondary outcomes were the VAS and SF-36 scores. Safety outcomes were the presence or absence of adverse events and side effects.

[0087] MDQ: The effect of OLL2809 on premenstrual and menstrual symptoms was investigated using the MDQ. The MDQ is a 46-item self-report questionnaire used to assess and treat premenstrual and menstrual symptoms. The 46 items are categorized into eight subfactors: (1) pain, (2) fluid retention, (3) autonomic nervous system changes, (4) negative affect, (5) poor concentration, (6) behavioral changes, (7) mood elevation, and (8) control. Of these subfactors, only (7) mood elevation captures positive changes.

[0088] For each symptom, "none" was assigned a score of 0, "slightly" a score of 1, "slightly strongly" a score of 2, "strongly" a score of 3, and "very strongly" a score of 4. The items for each subfactor were summed to obtain a subfactor score. The scores for (1) pain, (2) fluid retention, (3) autonomic nervous system changes, and (8) control were summed to obtain the "physical symptom score." The scores for (4) negative emotions, (5) decreased concentration, and (6) behavioral changes were summed and subtracted from this the score for (7) mood elevation to obtain the "mental symptom score." The mental symptom score and physical symptom score were summed to obtain an overall score.

[0089] SF-36: The SF-36 was used to evaluate health-related QOL. The answer sheets were processed using the WEB scoring program of Qualitest Co., Ltd., and the eight subscales of (1) physical functioning, (2) role functioning (physical), (3) bodily pain, (4) general health, (5) vitality, (6) social functioning, (7) role functioning (mental), and (8) mental health were scored on a scale of 0 to 100. Furthermore, these subscales were divided into physical health, mental health, and role / social health and scored accordingly. The higher the score, the better the health status.

[0090] VAS: From seven days before the expected onset of menstruation to the fourth day of menstruation, subjects recorded the severity of five symptoms, (1) lower abdominal pain, (2) rough skin, (3) swelling, (4) irritability, and (5) depression, once a day on a 100 mm scale ranging from 0 (no symptoms) to 100 (worst symptoms experienced). The maximum value for the seven days from seven days before menstruation to the day before menstruation was defined as the "premenstrual" value, and the maximum value for the four days from the first day of menstruation to the fourth day of menstruation was defined as the "menstrual" value. However, since the start date of VAS responses is the predicted date and the survey period fluctuates, if the premenstrual survey period was less than three days, it was considered missing.

[0091] 1-6. Target number of subjects In a previous study (Non-Patent Document 2) targeting endometriosis patients, the mean ± standard deviation of the change in menstrual pain VAS from the start of intake to 12 weeks after intake was -2.00 ± 1.67 in the placebo group (n = 33) and -3.28 ± 1.94 in the active group (n = 29), indicating a significant improvement (P < 0.01). In this study, the effect size (Hedges'g) of OLL2809 on menstrual pain VAS was 0.71. From this result, assuming that the effect size of OLL2809 on the MDQ score in this study is 0.7, the significance level is 5% and the power of detection is 80%, and the number of subjects per group is 33, so the required number of subjects was calculated to be 66. Considering the number of cases excluded from the efficacy analysis and the dropout rate of about 10%, the target number of subjects was set to 80.

[0092] Randomization The person responsible for allocation was someone who had no direct contact with on-site staff or the subjects. Five groups were created with subjects who had similar menstrual periods, and each group was randomly assigned to one of two groups - a test food intake group or a control food intake group - based on age, premenstrual MDQ total score before intake, and MDQ total score during menstruation.

[0093] 1-8. Blinding The Ethics Committee of Fukuda Internal Medicine Clinic, a medical corporation, conducted an indistinguishability test on the test foods and confirmed that the test foods and control foods were indistinguishable. The blinding of this study was maintained for all involved parties except the randomization manager by having the randomization list and test food identification code correspondence list, which were sealed immediately after allocation by the randomization manager, be kept securely until they were opened.

[0094] 1-9.Statistical analysis Data are presented as means and standard deviations (SD) or standard errors (SE). Statistical analysis was performed using SPSS version 26 (IBM). All tests were two-sided, with a significance level of 5%. Intergroup comparisons of subject background were performed using two-sample independent t-tests or Mann-Whitney U tests for quantitative data, and Fisher's exact test or Pearson's χ test for count data. 2 A multiple regression analysis was used to evaluate efficacy, with the change from before intake to the third menstrual cycle as the objective variable. The explanatory variables were the group (placebo group = 0, OLL2809 group = 1), analgesic use score before intake, stress score, and regular exercise habit at least once a week (no = 0, yes = 1). These were adopted as explanatory variables because there was a significant difference in the analgesic use score between the two groups in this study, and previous studies have shown that stress and exercise habits may affect menstrual symptoms (Non-Patent Documents 3, 8). All of these explanatory variables had variance inflation factors (VIFs) of 10 or less, and there were no problems with multicollinearity between explanatory variables. The estimated values ​​of the adjusted intergroup differences were shown with 95% confidence intervals (CI). In other analyses, an independent two-sample t-test was used for intergroup comparisons, and a paired t-test was used for pre- and post-intake comparisons.

[0095] 2.Results 2-1. Subject selection Consent to participate in the study was obtained from 207 subjects. As 9 subjects dropped out for personal reasons, the primary screening test was conducted on 198 subjects. Secondary screening was conducted on 120 subjects, and 80 subjects were enrolled in the study. The subjects were allocated to two groups of 40 subjects each. One subject in the OLL2809 group (for health reasons unrelated to the study) and two subjects in the placebo group (one subject was ineligible after the start of the study due to starting to take medicines that may affect the study, and one subject was seriously non-compliant with the study protocol) were discontinued at the discretion of the principal investigator, so the subjects analyzed for efficacy evaluation were 39 subjects in the OLL2809 group and 38 subjects in the placebo group (Figure 2). No cases were excluded from the analysis due to deviations from the study protocol.

[0096] There were no significant differences in the background characteristics of the OLL2809 group and the placebo group in terms of age, BMI, age at menarche, menstrual cycle, menstrual period, daily alcohol intake, simple stress score, premenstrual, menstrual and postmenstrual MDQ scores, neuroticism discrimination diagram area based on the CMI health questionnaire, smoking status, regular exercise habit at least once a week, and premenstrual analgesic use score (Table 1). On the other hand, the analgesic use score during menstruation was significantly higher in the placebo group than in the OLL2809 group.

[0097] The mean test food intake rates were 97.8% in the OLL2809 group and 98.3% in the placebo group, with no significant difference between the groups.

[0098] [Table 2]

[0099] 2-2.Effects of OLL2809 on premenstrual symptoms Table 2 shows the premenstrual MDQ scores. In the MDQ subfactors, the change in the "arousal" score from pre-intake in the OLL2809 group after the intervention for three menstrual cycles was significantly higher than that in the placebo group (Figure 3). Further analysis of the symptoms that influenced the "arousal" factor revealed that the item "Bursts of energy, activity" was significantly higher in the OLL2809 group than in the placebo group (Figure 4). Although no significant differences were observed between the groups in the items "Affectionate," "Orderliness," "Excitement," and "Feelings of well-being," the OLL2809 group showed higher scores.

[0100] The premenstrual SF-36 and VAS results are shown in Table 3. In the VAS evaluation of "irritability," the change in the OLL2809 group after three menstrual cycles was significantly lower than that in the placebo group (Figure 5). In the evaluation of "depressed mood" after three menstrual cycles, the change in the OLL2809 group showed a tendency to decrease compared to the placebo group (P = 0.070).

[0101] [Table 3]

[0102] [Table 4]

[0103] 2-3.Effects of OLL2809 on menstrual symptoms Regarding the MDQ scores during menstruation, no significant differences were found between the groups in the total score, physical symptom score, or mental symptom score.Also, no significant differences were found between the groups in the MDQ subfactors.Regarding the VAS scores during menstruation and the SF-36, no significant differences were found between the groups in any of the items.

[0104] 2-4. Safety evaluation of OLL2809 In this study, adverse events were defined as any undesirable or unintended injury, illness, or symptom thereof that occurred between the start of intake of the test food and the end of the study. No serious adverse events were identified in this study. During the observation period for adverse events (from the fifth day of menstruation in the 0th sexual cycle to the fourth day of menstruation in the 3rd sexual cycle), 34 adverse events were newly identified in the OLL2809 group and 34 in the placebo group, with no difference in incidence between the two groups. The causes of all adverse events that occurred were clear, and they were considered to be transient or accidental, and therefore were determined to be unrelated to intake of the test food.

[0105] 3. Discussion In Japanese women, the use of analgesics is the most common way to deal with menstrual pain. In this study, we excluded subjects who had a habit of taking analgesics prophylactically before the onset of menstrual pain, and instructed subjects not to take analgesics prophylactically before symptoms appeared, so that the use of analgesics would have as little effect on the evaluation of menstrual symptoms as possible. However, from the viewpoint of protecting subjects, we did not restrict the use of analgesics when symptoms appeared. Therefore, it was predicted that the use of analgesics would be a major confounding factor, and in fact, a significant difference was observed between the groups in the analgesic use score in the pre-intake period (Table 1). Therefore, we performed a multiple regression analysis using the analgesic use score as an explanatory variable along with other factors that affect menstrual symptoms. As a result, the OLL2809 group showed significantly higher scores than the placebo group in the MDQ "elevation of mood" factor before menstruation after the intervention for three menstrual cycles, which was the primary evaluation item. In particular, it was revealed that OLL2809 intake significantly improved the decrease in premenstrual vitality. In addition, the secondary endpoint of "irritability" was assessed using a visual analog scale (VAS), and the premenstrual score was significantly lower in the OLL2809 group compared to the placebo group. These results indicate that the intake of OLL2809 reduces premenstrual decline in vitality and irritability, and improves menstrual symptoms.

[0106] In this study, OLL2809 was found to improve premenstrual mental symptoms, but no improvement was observed in premenstrual physical symptoms or menstrual symptoms under the experimental conditions. There have been many reports on the relationship between physical and mental symptoms associated with menstruation, and it has been said that headaches and migraines associated with neurovascular disorders due to estrogen withdrawal before menstruation and lower abdominal pain associated with uterine contractions induced by prostaglandins affect mental symptoms such as depression and irritability (Non-Patent Document 10). In a report examining the relationship between physical symptoms of the menstrual cycle and premenstrual depressive symptoms, a strong linear effect and a moderate curvilinear effect were observed when physical symptom scores were used as a predictor of depressive symptoms. When the level of physical symptoms was low, no association was observed between physical symptoms and depressive symptoms, but the association became stronger as the level of physical symptoms increased (Non-Patent Document 9). It is possible that OLL2809's effect in improving menstrual symptoms is exerted before menstruation, when the level of physical symptoms is relatively low, and that it did not have any effect on the physical symptoms and the mental symptoms affected by them during menstruation, when menstrual pain (lower abdominal pain, headache) symptoms become more severe than before menstruation.

[0107] 4. Conclusion In this study, a randomized, placebo-controlled, double-blind, parallel-group comparative study was conducted on healthy women who were aware of menstrual symptoms, using OLL2809 as the test food. Since the subjects were healthy adult women, subjects with diseases such as PMS, PMDD, dysmenorrhea, or endometriosis, and subjects who were judged to be neurotic were excluded. In addition, subjects with symptoms severe enough to interfere with daily life were also excluded because they may have been caused by the above-mentioned diseases. Therefore, the results of this study are considered to be meaningful in the sense that they can provide a way of dealing with mental disorders before menstruation other than medication for women who are not serious enough to go to the hospital. From the above, it was confirmed that the intake of OLL2809 for three menstrual cycles improves the decrease in vitality and irritability before menstruation in healthy women, suggesting that OLL2809 may contribute to improving women's QOL.

[0108] 5. References cited in the specification (Non-patent document 2) Itoh H, Uchida M, Sashihara T, et al. Lactobacillus gasseri OLL2809 is effective especially on the menstrual pain and dysmenorrhea in endometriosis patients: randomized, double-blind, placebo-controlled study. Cytotechnology 2011: 63: 153-161. (Non-Patent Document 3) Matsumoto T, Egawa M, Kimura T, Hayashi T. A potential relation between premenstrual symptoms and subjective perception of health and stress among college students: a cross-sectional study. BioPsychoSocial Medicine 2019: 13: 26. (Non-Patent Document 4) Moos RH. The development of a menstrual distress questionnaire. Psychosom Med 1968: 30: 853-867. (Non-Patent Document 5) Ken Fukamachi. Study of the Cornell Medical Index (Part 2) Criteria for distinguishing neurotic patients using the CMI. Fukuoka Medical Journal 1959: 50: 3001-3009. (Non-Patent Document 6) Murakami, Masato. Research on the stress state of healthy people, how symptoms caused by stress appear and how to deal with them. Psychosomatic Medicine 1989: 1: 72-82. (Non-Patent Document 7) Fukuhara S, Bito S, Green J, Hsiao A, Kurokawa K. Translation, adaptation, and validation of the SF-36 Health Survey for use in Japan. J Clin Epidemiol 1998: 51: 1037-1044. (Non-patent document 8) Mizuta R, Maeda N, Komiya M, et al. The relationship between the severity of perimenstrual symptoms and a regular exercise habit in Japanese young women: a cross-sectional online survey. BMC Womens Health 2022: 22: 200. (Non-patent document 9) Kiesner J. Physical characteristics of the menstrual cycle and premenstrual depressive symptoms. Psychol Sci 2009: 20: 763-770. (Non-Patent Document 10) Kiesner J, Eisenlohr-Moul T, Mendle J. Evolution, the Menstrual Cycle, and Theoretical Overreach. Perspect Psychol Sci 2020: 15: 1113-1130. [Industrial Applicability]

[0109] The present invention supports the maintenance and improvement of people's health by providing a composition for treating menstrual symptoms, which contains lactic acid bacteria belonging to Lactobacillus paragasseri. The present invention also provides a food composition and a method for producing a food that support the maintenance and improvement of people's health. Furthermore, the present invention can improve the nutrition of various people, ensure healthy lives, and promote welfare.

[0110] [Sequences listed in the sequence listing] SEQ ID NO:1 Lactobacillus paragasseri OLL2809 16s rRNA gene

Claims

1. A composition for treating menstrual symptoms, comprising lactic acid bacteria belonging to Lactobacillus paragasseri.

2. The composition according to claim 1, wherein the menstrual symptoms are either premenstrual physical discomfort or premenstrual mental discomfort.

3. The composition according to claim 1, wherein the menstrual symptoms are premenstrual mental discomfort symptoms.

4. 10. The composition of claim 1 for ingestion by a subject not suffering from menopausal symptoms or by a subject in adolescence or sexual maturity.

5. The composition according to claim 1, wherein the menstrual symptoms are mental discomfort symptoms, and the mental discomfort symptoms are one or more selected from irritability, depression (low mood), intolerance, irritability, confusion, emotional instability, dissatisfaction, hopelessness, loss of energy, fatigue, and loss of motivation.

6. A composition for controlling premenstrual highs comprising Lactobacillus paragasseri.

7. A composition for controlling premenstrual irritability comprising Lactobacillus paragasseri.

8. The composition according to claim 1 , comprising Lactobacillus paragasseri as heat-induced bacterial cells.

9. The composition according to any one of claims 1 to 7, comprising only Lactobacillus paragasseri cells as an active ingredient.

10. The composition according to any one of claims 1 to 7, wherein Lactobacillus paragasseri is Lactobacillus paragasseri OLL2809 deposited under accession number NITE BP-72, or a strain taxonomically equivalent thereto.

11. The daily dose of Lactobacillus paragasseri cells was 1 × 10 8 ~1×10 12 A composition according to any one of claims 1 to 7 for individual ingestion.

12. A method (excluding medical procedures) for treating symptoms associated with menstruation, comprising a step of having a subject ingest a composition containing lactic acid bacteria belonging to Lactobacillus paragasseri.

13. The daily dose of Lactobacillus paragasseri cells was 1 × 10 8 ~1×10 12 The method according to claim 12, wherein the individual is ingested.

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