JP5873904B1 - Composition for improving restless legs syndrome - Google Patents

Abstract

[PROBLEMS] All drugs such as drugs that improve the symptoms of restless legs syndrome have known side effects, and some renal excretion type restless legs syndrome worsens the symptoms of patients with renal dysfunction It's hard to say it's safe for you. An object of the present invention is to provide a composition that exhibits an effect of improving restless legs while being a food that is safe and has no side effects. [MEANS FOR SOLVING PROBLEMS] It has been found that lactic acid bacteria or cultures inoculated with lactic acid bacteria have an effect of promoting dopamine secretion, and further confirmed that they have a high effect of improving restless legs syndrome and have very few side effects. The invention has been completed. In the present invention, when the lactic acid bacterium is Lactobacillus crispatus KT-11 strain (FERM BP-11332), various functions are expressed more effectively. [Selection figure] None

Description

  The present invention relates to a composition having very few side effects for improving restless legs syndrome comprising a lactic acid bacterium or a culture inoculated with lactic acid bacteria and a food containing the same.

  Restless Legs Syndrome, also known as Willis Echbom's disease, restless leg syndrome, has traditionally been called restless leg syndrome. Restless legs syndrome is a common disease affecting about 10% of people in the West and 2% to 5% in Japan. Symptoms of restless legs syndrome appear mainly in the lower limbs, and they feel itching, crawling, ugly, and painful. These symptoms sometimes appear in the whole body, such as in the lower back and arms. Symptoms may be temporarily relieved by exercise or gestures, but the patient has an unbearable urge to move the lower limbs and body. Symptoms occur in particular from evening to night, but they worsen during daytime rest, making it impossible to take a steady posture while sitting on a chair, and patients may have trouble in daily life and work.

  The causes of restless legs syndrome are known to be iron deficiency and a secretion disorder in the center of dopamine, which is a neurotransmitter (Non-patent Document 1 J Neurol 2009; 256: 539-553).

  Therefore, dopamine agonists (pramipexole hydrochloride, ropinirole, talipexol, cabergoline) and levodopa preparations that are dopamine agonists are effective as therapeutic drugs for restless legs syndrome (Patent Document 1). In addition, drugs such as benzodiazepine sedatives, GABA inducers (Patent Document 2), opioids, iron materials, adenosine A2A receptor antagonists (Patent Document 3) are used for treatment.

  However, these drugs include nausea, sudden sleep, antidiuretic hormone incompatible secretion syndrome, constipation, hallucinations, delusions, delirium, agitation, confusion, malignant syndrome, interstitial pneumonia, pleurisy, pleural effusion, pleural fibrosis, pulmonary fiber There are problems that are not necessarily safe, such as known side effects such as symptom, pericarditis, pericardial effusion, valvular heart disease, rhabdomyolysis.

  By promoting the release of dopamine in the central nervous system with drugs, restless legs syndrome can be improved. However, given the possibility of serious side effects, it can be safely administered without the risk of side effects. It is desirable that the composition be available.

  However, no ingredient or food having an effect of improving restless legs syndrome derived from natural products has been reported so far. Furthermore, there is no known composition for improving restless legs syndrome, which contains a lactic acid bacterium or a culture inoculated with lactic acid bacteria as an active ingredient.

Further Lactobacillus crispatus of the present invention, wherein (Lactobacillus crispatus) KT-11 strain (FERM BP-11332) has been found to have anti-allergic effect be taken orally (Patent Document 4) It was not known at all to have an effect of improving restless legs syndrome.

JP2014-074013 Patent 4216713 Patent 4498140 Patent 4921499

Trenkwalder C. et al. “J. Neurol.” 2009, 256, p539-553.

  Drugs that improve symptoms of restless legs syndrome have known side effects, and some renal excretion-type drugs that improve restless legs syndrome are safe for all patients, such as worsening the symptoms of patients with renal dysfunction It is hard to say that there is.

  An object of the present invention is to provide a composition that exhibits a restless legs improving effect while being a food that is safe and has no side effects.

  As a result of earnest research to achieve the above-mentioned problems, we found that cultures inoculated and inoculated with lactic acid bacteria or lactic acid bacteria have a dopamine secretion promoting action, and further improved high restless legs syndrome The present invention was completed by confirming that it was effective and had very few side effects.

  The composition for improving restless legs syndrome of the present invention contains, as an active ingredient, a natural product obtained by inoculating lactic acid bacteria or inoculating lactic acid bacteria. Therefore, the symptoms of restless legs syndrome can be reduced without worsening the symptoms of patients with side effects and renal dysfunction like conventional drugs. Moreover, the symptom of restless legs syndrome can also be reduced by ingesting the foodstuff containing this composition.

  The composition of the present invention comprises a culture obtained by inoculating and cultivating lactic acid bacteria in a medium containing lactic acid bacteria or a culture substrate. The culture may be any of live and dead cells, and any state such as dry cells, wet cells, culture solution, cell suspension, cell disruption, cell components, etc. It may be used, and may be a culture filtrate from which the cells are removed. Moreover, as a form of the culture, any form such as a solid, a powder, a granule, a paste, and a liquid may be used.

  The lactic acid bacteria can be cultured under any conditions according to a conventional method. The culture method is not particularly limited, and a conventionally known method can be appropriately selected and used. For example, it can be performed under aerobic conditions or anaerobic conditions. In general, the culture temperature is preferably 30 to 37 ° C., but may be other temperature conditions as long as the bacterium grows. Although it is desirable to maintain the pH of the medium during the culture at 5.0 to 7.0, other pH conditions may be used as long as the pH allows the bacteria to grow. The culture time is usually preferably 12 to 36 hours, but any other culture time may be used as long as the bacteria can grow.

  As the medium for culturing lactic acid bacteria, any medium can be used as long as it contains moderately carbon sources and other nutrients for promoting growth. For example, MRS medium, GYP medium, BL medium, GAM medium, and tomato juice medium are used. Moreover, barley extract, soybean extract, and milk are used for the culture base material used for a culture medium. Examples of barley extract include fermented barley extract, barley enzyme hydrolyzate, malt extract, barley young leaf extract, and barley shochu. Examples of soybean extract include defatted soybean powder, soy milk, and soybean enzyme hydrolyzate. Examples of milk include skim milk powder, enzyme hydrolyzed milk, casein, and whey. Of these medium base materials, barley extract is most effective as a medium for culturing lactic acid bacteria. In the present invention, among the components constituting the medium, the main component excluding moisture is defined as the culture substrate.

  As the carbon source, glucose, lactose, sucrose, fructose, starch hydrolyzate, molasses, etc. can be used. In addition, meat extract, fish extract, yeast extract, oleic acid and the like are used as nutrients.

A strain of lactic acid bacteria can be used in the present invention include Lactobacillus crispatus is a human-derived lactic acid bacteria (Lactobacillus crispatus) KT-11 strain (FERM BP-11332). The strain is deposited with the Patent Organism Depositary, National Institute of Advanced Industrial Science and Technology. As described in Patent Document 4 (Patent 4921499), the mycological (morphological, cultivating, physiological) properties of the strain are as follows.

[1] Morphological properties
In observation after culturing at 37 ° C. for 72 hours on MRS agar medium (Nippon Becton Dickinson), the cell size is 0.5-1 × 3-5 μm bacilli and does not move. There is no sporulation and Gram staining is positive.
[2] Culture characteristics
Colonies after culturing at 37 ° C. for 72 hours on MRS agar medium (Nippon Becton Dickinson) have a diameter of 2 to 3 mm, are round, and have full edge. The color of the colony is yellowish white and translucent.
[3] Physiological properties Gas production negative Glucose utilization positive Catalase activity negative Gelatin liquefaction negative Nitrate reduction negative Indole production negative Hydrogen sulfide production negative Attitude to oxygen Facultative anaerobic Optimal growth temperature 37-40 ° C
Optimum growth pH pH 5.5-5.8

The present invention, when lactic acid bacteria as can be seen from Figure 1 is the above Lactobacillus crispatus (Lactobacillus crispatus) KT-11 strain (FERM BP-11332), the amount of dopamine production lot increases than compared with other lactic acid bacteria, Various functions are expressed more effectively.

Further, lactic acid bacteria as necessary, in addition to Lactobacillus of the deposited strains (Lactobacillus), Lactococcus (Lactococcus), Streptococcus (Streptococcus), Enterococcus (Enterococcus), Pediococcus (Pediococcus), Leuconostoc (Leuconostoc) Any lactic acid bacteria belonging to Bifidobacterium can be used without any particular limitation.

For example, Lactobacillus acidophilus , Lactobacillus casei , Lactobacillus gasseri , Lactobacillus fermentum , Lactobacillus fermentum , Lactobacillus plantarum Bacillus brevis (Lactobacillus brevis), Lactobacillus reuteri (Lactobacillus reuteri), Lactobacillus crispatus (Lactobacillus crispatus), Lactobacillus del Burukki subsp. bulgaricus (Lactobacillus delbrueckii subsp. bulgaricus), Lactobacillus John Sony (Lactobacillus johnsonii), Lactococcus lactis subsp. lactis (Lactococcus lactis subsp. lactis), Lactococcus lactis subsp. cremoris (Lactococcus lactis su bsp. cremoris), Streptococcus thermophilus (Streptococcus thermophilus), Enterococcus faecalis (Enterococcus faecalis), Enterococcus faecium (Enterococcus faecium), Pediococcus pentosus (Pediococcus pentosaceus), Pediococcus reeds di Lacty Shi (Pediococcus One or two or more lactic acid bacteria selected from acidilactici ), Leuconostoc mesenteroides Bifidobacterium bifidum, and the like may be used.

  Moreover, the composition for restless legs syndrome improvement of this invention can be mix | blended with a foodstuff. In the present invention, the food may be used in the sense of including health food, functional food, nutritional supplement, or food for specified health use. Furthermore, when the food of the present invention is used for mammals other than humans, it can be used in the meaning including pet food, supplements for pets, feed, and the like.

  Specific examples of food include various beverages such as juices, soft drinks, tea beverages, drinks, jelly-like beverages and functional beverages, alcoholic beverages such as beer, carbohydrates such as cooked rice, noodles, breads and pasta Contained food, fish paste products such as kamaboko and chikuwa, livestock processed products such as ham and sausage, retort products such as curry, sauce, and Chinese soup, soups, dairy products such as milk, milk drinks, ice cream, cheese, and yogurt, Fermented products such as miso, yogurt, lactic acid bacteria, fermented beverages, pickles, Western confectionery such as bean products, biscuits and cookies, Japanese confectionery such as buns and sheepskin, frozen confectionery such as candies, gums, gummy, jelly, pudding Confectionery such as instant food such as instant soup and instant miso soup, food for microwave oven, mayonnaise and dressy Gu seasoned seasoning liquid, such as seasonings, dog food, and a pet food cat food, and the like. Furthermore, health foods and drinks prepared in the form of powder, granules, tablets, capsules, liquid, paste or jelly are also included.

  The composition for improving restless legs syndrome of the present invention can be formulated and used by mixing with a physiologically acceptable carrier, excipient, binder, diluent and the like. The composition for improving restless legs syndrome can be administered orally or parenterally, but is preferably administered orally. Oral preparations include granules, powders, tablets (including sugar-coated tablets), pills, capsules, syrups, emulsions, and suspensions. Examples of parenteral preparations include external preparations (eg, nasal preparation, transdermal preparation, ointment) and suppositories (eg, rectal suppository, vaginal suppository).

  The daily intake of the composition for improving restless legs syndrome of the present invention varies depending on age, symptoms, body weight, use, etc., and thus cannot be determined in general, but as a guideline, 0.1 to 1000 mg / kg body weight Is preferable.

  According to the present invention, a composition for improving restless legs syndrome with very few side effects is provided by ingesting as a food a culture composition obtained by inoculating and cultivating lactic acid bacteria in a lactic acid bacterium, preferably a barley extract.

The figure which showed the amount of dopamine production in the culture supernatant of PC12 cell cultured by adding lactic acid bacteria or lactic acid bacteria culture. The figure which shows the improvement of the symptom of the restless legs syndrome by ingestion of the composition of this invention.

  An example of a method for obtaining a composition for improving restless legs syndrome used in the present invention and an effect thereof will be described, but the present invention is not particularly limited thereto.

[Example 1] Production of lactic acid bacteria or lactic acid bacteria culture The medium used in the present invention is not particularly limited. For example, MRS medium (Nippon Becton Dickinson), barley extract (Sanwa) are used as the culture substrate. Liquor), defatted soybean flour (Nippon Becton Dickinson), skim milk (Wako Pure Chemicals), etc. can be used. The composition of the medium is shown in Table 1.

Lactic acid bacteria are Lactobacillus acidophilus , which is an animal lactic acid bacterium, Lactobacillus plantarum , which is a plant lactic acid bacterium, Bifidobacterium bifidum , which is a Bifidobacterium, and lactic acid cocci. An Enterococcus faecalis was used. Lactic acid bacteria were inoculated in the medium shown in Table 1 and cultured at 37 ° C. for 24 hours.

  For the preparation of lactic acid bacteria, the cells were collected by centrifugation (2,000 rpm, 10 minutes) after culturing, and the medium components were completely removed by centrifugal washing with purified water three times. Washed lactic acid bacteria are heat-treated (110 ° C, 15 minutes), lyophilized, and then suspended in sterile 0.15 M sodium chloride-0.01 M phosphate buffer (PBS, pH 7.2). did.

  Preparation of lactic acid bacteria culture is heat treatment after incubation (110 ° C, 15 minutes), lyophilized, and then suspended in sterile 0.15 M sodium chloride-0.01 M phosphate buffer (PBS, pH 7.2). It was.

[Example 2] Induction test of dopamine production by lactic acid bacteria [Brown cell culture]
PC12 cells (rat adrenal medulla-derived pheochromocytoma: RIKEN Cell Bank), a mouse brown cell, is a DMEM medium (Nacalai) containing 1% FBS, 1% HS, 100 U / mL penicillin G sodium and 100 μg / mL streptomycin sulfate. ) And then cultured in a sterile plastic petri dish at 37 ° C. in the presence of 5% CO ₂ until confluence. Lactic acid bacteria or lactic acid bacteria suspended in PBS to a final concentration of 0 or 1000 μg / mL in a 48-well flat-bottom microplate dispensed with 1 mL of PC12 cell suspension prepared to 1 × 10 ⁶ cells / mL 100 μL of each culture was added to each well and cultured at 37 ° C. in the presence of 5% CO ₂ for 24 hours. Dexamethasone was used as a positive control. After culturing, centrifugation was performed at 4 ° C. and 2000 rpm for 15 minutes, and the culture supernatant was collected.

[Dopamine production]
Dopamine production was measured according to an enzyme immunoassay (ELISA) kit (LDN, highly sensitive dopamine BA E-5300). The dopamine production amount in the PC12 cell culture supernatant was measured by measuring the absorbance at 450 nm using a Bio-Rad model 550 microplate reader. The dopamine production was calculated from a standard curve obtained from a known concentration of dopamine. Statistical significance was determined by Student's T-test based on dopamine production when no sample was added.

  FIG. 1 shows the amount of dopamine produced in the culture supernatant of PC12 cells cultured with lactic acid bacteria or lactic acid bacteria culture added. The amount of dopamine produced was significantly increased when all lactic acid bacteria or lactic acid bacteria cultures were added, compared to the case where no lactic acid bacteria were added, and the dopamine production was increased more in the culture than in lactic acid bacteria. In particular, it was shown that dopamine production is significantly increased in lactic acid bacteria cultures using barley extract as a culture substrate (medium B in Table 1).

[Example 3] Human ingestion test of lactic acid bacteria culture In order to confirm the improvement effect of restless legs syndrome by lactic acid bacteria culture, a preparation of lactic acid bacteria culture was prepared and administered to patients suffering from restless legs syndrome .

[Production of lactic acid bacteria culture powder]
Lactobacillus crispatus (Lactobacillus crispatus) KT-11 strain (FERM BP-11332), 37 ℃ in a medium shown in Table 1, and cultured for 24 hours. After culturing, the mixture was sterilized by heating at 110 ° C. for 10 minutes, neutralized with calcium carbonate to pH 5.8, mixed with dextrin, and lyophilized to prepare a barley extract lactic acid bacteria culture powder.

[Production method]
Lactic acid bacteria culture powder, crystalline cellulose, reduced maltose starch syrup, and sucrose fatty acid ester were mixed and tableted with a single tableting machine to produce tablets with a diameter of 9 mm and a weight of 350 mg. The obtained tablet was sprayed with a shellac solution and then dried to produce a tablet containing a culture of lactic acid bacteria coated with shellac. This composition is shown in Table 2.

[Evaluation of symptom improvement in patients with restless legs syndrome]
For the test, 10 Japanese female volunteers aged 20 to 59 who suffer from restless legs syndrome were selected. Tablets containing the barley extract lactic acid bacteria culture shown in Table 2 were ingested daily for 6 months, 6 tablets per day. Symptoms of restless legs syndrome were assessed using a visual rating scale (VAS) for leg discomfort, effects on daily life, desire to move around, and general severity of symptoms.

  VAS is “0” for “no pain” and “10” for “no more pain”. It is a method to show. The result is shown in FIG.

  From the results shown in FIG. 2, there was a statistically significant improvement effect in the items of unpleasant sensation of the leg, influence on daily life, desire to move around, and severity of general symptoms after intake compared to before intake. It was.

  In addition, no side effects such as nausea observed with drugs for restless legs syndrome were observed, and safety was also confirmed.

Claims (3)

Lactobacillus chryspatus, Lactobacillus acidophilus, Lactobacillus plantarum, Bifidobacterium bifidum or Enterococcus faecalis cells or a culture of the fungus body as an active ingredient A composition for improving restless legs syndrome. The Lactobacillus crispatus is Lactobacillus crispatus (Lactobacillus crispatus) KT-11 strain (FERM BP-11332) for improving composition according to claim 1 of the restless legs syndrome, a. The composition for improving restless legs syndrome according to claim 1 or 2, wherein the culture substrate of the culture of the bacterial cells is barley extract.

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