JP5172104B2 - Bacteria-containing composition with leprosy prevention effect - Google Patents

Abstract

Disclosed is a composition comprising at least one Gram-positive bacterium isolated from the intestine of an animal including human or a fermented food as an active ingredient. The composition can be used for inhibition or reduction of in vivo proliferation of Mycobacterium leprae. The composition can be used for prevention, progression prevention or amelioration of Hansen's disease or a condition associated with Hansen's disease. The composition can be ingested readily in a daily life, is safe, and develops reduced adverse side effects.

Description

Background of the Invention

FIELD OF THE INVENTION The present invention relates to a fungus body-containing composition having an effect of preventing the onset of Hansen's disease. More particularly, the present invention relates to a composition comprising a bacterium selected from the group consisting of gram positive bacteria isolated from animal intestines including humans or from fermented foods. Moreover, this invention relates to the food-drinks which have the leprosy prevention effect.

Background Art Leprosy is one of infectious diseases mainly affected by skin and peripheral nerves by Mycobacterium leprae . In Japan, about 10 cases or less are reported annually, but it is said that there are still millions of people with leprosy in developing countries.

  For example, in Indonesia, India, and Africa, there are 3 to 4 people per 10,000 people, and there are 10 times more people with leprosy in areas with a high prevalence. Such areas are said to be leprosy-contaminated areas. In these areas, the number of new patients infected at an early age due to infection within the family increases, and measures to prevent the onset of leprosy are needed. For the treatment of leprosy, multi-drug combination therapy (MDT) has been effective, but no effective means for preventing infection has yet been found for its onset prevention. For this reason, the increase in newly infected people remains serious. Furthermore, recently, there are concerns about the development of leprosy having multidrug resistance, and it is important to find infection prevention measures that are not related to conventional drugs.

  Furthermore, from the viewpoint of preventing the onset, it is desired to provide a drug or food that has fewer side effects and can be used more safely. Providing such drugs or foods is expected to help improve the quality of life (QOL) of patients.

  On the other hand, in recent years, research on intestinal flora has progressed, and it has become clear that intestinal flora is closely related to the health and diseases of the host, and lactic acid bacteria and bifidobacteria have been used in various forms such as probiotics. There are many attempts to use it to prevent disease and improve symptoms.

  For example, Japanese Unexamined Patent Publication No. 2000-125810 (Patent Document 1) discloses a concentrated beverage having an anti-inflammatory action, which comprises yeast and lactic acid bacteria. JP-A-11-259281 (Patent Document 2) discloses an immunomodulator using a dead cell of a specific genus Enterococcus.

Enterococcus faecalis EC-12 (hereinafter referred to as “EC-12 strain”), which is one of lactic acid bacteria, has an intestinal action and antitumor effect on its dead cells. It has been reported that there are effects, skin allergy improving action, etc. (for example, “Food and Development”, Vol. 39, No. 10, pp. 61-63 (Non-patent Document 1)). Regarding the EC-12 strain, there are also reports on the effect on the host defense mechanism during Listeria monocytogenes infection ("New Drug and Clinical" (J. New Rem. & Clin.), Vol. 53, No. 3, 2004, pp. 78-88 (Non-Patent Document 2)).

  However, as far as the present inventors know, nothing has been reported regarding the leprosy prevention effect of these bacteria. The infection and onset mechanism of leprosy is completely different from the onset mechanism of Listeria monocytogenes and various diseases described above.

JP 2000-125810 A Japanese Patent Laid-Open No. 11-259281 “Food and Development”, Vol. 39, No. 10, pp. 61-63 "New drugs and clinical practice", Vol. 53, No. 3, 2004, pp. 78-88

Summary of the Invention

  The present inventors are now able to remarkably suppress the growth of leprosy in the living body by ingesting EC-12 strain, which is one of Gram-positive bacteria isolated from animal intestines including humans or fermented foods, and leprosy. It was unexpectedly found that the symptoms associated with can be improved. The present invention is based on such knowledge.

  The object of the present invention is to prevent, progress, or improve leprosy or a condition related to leprosy that can be easily ingested in daily life and that can be used as a medicine or food and drink with very few side effects. It is to provide a composition.

  The composition according to the present invention comprises, as an active ingredient, at least one kind of Gram-positive bacteria isolated from animal intestines including humans or fermented foods, and is used to suppress or reduce the growth of leprosy in a living body. It is used.

  According to a preferred embodiment of the present invention, the bacterium is a dead cell.

  According to a preferred embodiment of the present invention, the composition according to the present invention is used for the prevention, progression inhibition or amelioration of leprosy or related conditions. More preferably, the composition according to the present invention prevents leprosy or related conditions in patients infected with or suspected of being infected with leprosy, or in an environment likely to be infected with leprosy. Used to suppress or improve progress.

  According to another aspect of the present invention, there is provided a food or drink comprising the composition according to the present invention.

  According to another aspect of the present invention, there is provided a food or drink comprising an effective amount of the active ingredient, which is used for inhibiting or reducing the growth of leprosy in a living body.

  According to still another aspect of the present invention, there is provided a food or drink comprising an effective amount of the active ingredient, wherein the food or drink is displayed with a function for preventing, suppressing progression or improving leprosy or a state related thereto. Is done.

  According to the present invention, growth of leprosy in a living body can be suppressed or reduced by ingesting an active ingredient as a medicine or a food or drink. That is, according to the present invention, leprosy or a state associated therewith can be prevented, progressed or improved by ingesting the active ingredient as a pharmaceutical or a food or drink. The active ingredient in the present invention is bacteria isolated from animal intestines including humans or fermented foods or dead cells thereof, and it is clear from previous reports that there are almost no side effects and high safety. For this reason, the composition or food-drinks by this invention have few side effects, and are excellent in safety. Therefore, they can be easily taken in daily life. Therefore, since the composition of the present invention can be easily taken by children and adults without side effects, it effectively suppresses leprosy family infection and can effectively prevent childhood infection and leprosy increase in leprosy-contaminated areas. It is expected.

Detailed description of the invention

Deposit of Microorganisms Enterococcus faecalis · EC-12 strain (Enterococcus faecalis EC-12), the 2005 (2005) February 25, National Institute of Advanced Industrial Science and Technology, International Patent Organism Depositary dated (original deposit date) Deposited at 1-6 Higashi 1-chome, Tsukuba City, Ibaraki Prefecture 305-5466, Japan. The accession number is FERM BP-10284.

Active ingredient The active ingredient of the composition and food / beverage products according to the present invention is, as described above, at least one Gram-positive bacterium isolated from animal intestines or fermented foods including humans. This bacterium is a gram-positive bacterium that produces lactic acid, a so-called lactic acid bacterium.

According to a preferred embodiment of the present invention, the bacterium is Enterococcus genus (Enterococcus), bifidobacteria (Bifidobacterium), Lactobacillus (Lactobacillus), Streptococcus (Streptococcus), and belong to the Lactococcus (Lactococcus) Selected from the group consisting of bacteria.

Here, examples of bacteria belonging to the genus Enterococcus include Enterococcus faecalis , Enterococcus faecium, and the like. Examples of bacteria belonging to the genus Bifidobacterium include Bifidobacterium longum , Bifidobacterium breve , and Bifidobacterium bifidum. . Examples of bacteria belonging to the genus Lactobacillus include Lactobacillus acidophilus , Lactobacillus casei , Lactobacillus salivarius and the like.

Examples of bacteria belonging to the genus Streptococcus include Streptococcus thermophilus . Examples of bacteria belonging to the genus Lactococcus include Lactococcus cremoris and Lactococcus lactis .

According to a preferred embodiment of the present invention, the bacterium used as an active ingredient is a bacterium belonging to the genus Enterococcus. More preferably, the bacterium is Enterococcus faecalis .

Examples of Enterococcus faecalis include strains such as Enterococcus faecalis EC-12 (accession number FERM BP-10284), ATCC 19433, ATCC 14508, ATCC 23655, IFO 16803, IFO 16804, and mutants thereof. Of these, the EC-12 strain is most preferred as the bacterium used as the active ingredient.
Here, the term “mutant strain” means that a specific strain has been mutated by a method well known to those skilled in the art within a range that does not change the properties thereof, or is equivalent to that. The meaning includes things that can be confirmed.

  Bacteria used as an active ingredient can be live bacteria and / or dead cells, preferably dead cells, more preferably obtained by sterilizing the bacteria with a known heat treatment means. Heat sterilized cells are used. The heat-sterilized bacterial cell is collected from the culture obtained by culturing the bacterium according to a conventional method, for example, by filtration, centrifugation, or the like, washed with water, suspended in water or the like and 120 ° C. or lower. (Preferably 80 to 120 ° C.) Heat treatment within 30 minutes (3 seconds to 30 minutes), followed by concentration and drying as necessary. Note that enterococcus faecalis and bactericidal cell powder by heat treatment of the EC-12 strain are commercially available under the trade name “EC-12” (manufactured by Combi Corporation). For this reason, in this invention, you may use such a commercial item as bacteria used as an active ingredient.

Use The active ingredient according to the present invention has the effect of suppressing or reducing the growth of leprosy in the living body. That is, as shown in the examples described later, the active ingredient according to the present invention suppresses the growth of larvae in a living body due to infection with bacterium by ingesting it to rats sensitive to bacterium infection. Is actually confirmed (Example, FIG. 1 and FIG. 2). That is, it was confirmed from the findings of the rats of the active ingredient administration group and the non-administration group according to the present invention that the progression of the disease state associated with the infection with bacterium was reduced by the composition according to the present invention. Thus, the active ingredient by this invention actually showed the effect which prevents and progresses the leprosy or the state related to this. Therefore, the active ingredient in the present invention has an effect of suppressing the progression of symptoms such as the reduction of symptoms such as the reduction of swelling, the mitigation of symptoms, and the improvement effect on patients infected with leprosy or those who are likely to be infected. It can also bring about a therapeutic effect on symptoms. Further, the active ingredient of the present invention has low toxicity and can be used safely for mammals.

  According to the active ingredient of the present invention, it is considered that the growth of leprosy, an intracellular parasitic bacterium, can be suppressed by the action of lactic acid cocci on the immune system, and as a result, the onset of leprosy can be prevented. These are assumptions, and the present invention is not construed as being limited thereto.

  Therefore, the active ingredient according to the present invention can be used for prevention, progression inhibition or improvement of leprosy or a condition related thereto. Here, the “progression suppression or improvement” of a symptom or condition is used in the meaning including regulation, suppression of progression, delay, alleviation, improvement, prevention of re-progression of symptoms, etc.

In the present invention, “leprosy” refers to an infection caused by infection with Mycobacterium leprae , which includes depigmentation or red rash with sensory escape, peripheral nerve thickening, annular erythema caused by infection with Leprosy. And symptoms such as leprosy reactions.

  The effect of the active ingredient according to the present invention can be confirmed by measuring the degree of swelling of the limbs, the thickness of the foot pads of the hind limbs, etc. over time.

  According to another aspect of the present invention, a leprosy growth inhibitor comprising at least one Gram-positive bacterium isolated from animal intestines including humans or fermented food as an active ingredient, or leprosy or A preventive agent, progress inhibitor or ameliorating agent for a condition related to this is provided.

  Further, according to the present invention, as described above, a patient who is infected or suspected of being infected with leprosy, or a person who is placed in an environment where there is a high possibility of being infected with leprosy, or who develops or is suspected to develop leprosy. Preventing, suppressing or improving the progression of leprosy or related conditions comprising administering or ingesting an effective amount of the active ingredient to a patient with A method is provided. As used herein, “effective amount” means the amount of an active ingredient that is required at least to exert its effect in order to suppress or improve the progression of symptoms or conditions. The “patient” refers to a human or a mammal other than a human to be administered or ingested the composition or food or drink of the present invention (in particular, immunodeficient mice, rats, and armadillos that are sensitive to leprosy). Monkeys). “Administration” and “intake” mean that a target substance is taken into a living body orally or parenterally (preferably orally) to a subject patient.

Composition or food or drink The composition according to the present invention comprises the above-mentioned active ingredient.

  As used herein, “comprising as an active ingredient” may contain a physiologically acceptable carrier according to the desired product form, and may contain other auxiliary ingredients that can be used in combination. It means to include. That is, the composition according to the present invention is mixed with physiologically acceptable carriers, excipients, binders, diluents, etc., if necessary, using bacterial cells or dead cells which are active ingredients. Can be manufactured. The composition according to the present invention can be administered or ingested orally or parenterally. Examples of oral forms include foods, granules, powders, tablets (including sugar-coated tablets), pills, capsules, syrups, emulsions, and suspensions. Examples of parenteral forms include injections, drops, and external preparations. These preparations can be formulated together with a pharmaceutically acceptable carrier (for example, an excipient or an additive) by a method commonly used in the art. Pharmaceutically acceptable carriers include excipients, binders, fragrances, buffers, thickeners, colorants, stabilizers, emulsifiers, dispersants, suspending agents, disintegrants, lubricants, preservatives. Agents and the like. Specific examples include magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, a low melting point wax, and the like.

  Other auxiliary components that can be used in combination include, for example, vitamin components (eg, vitamin C, vitamin E), antibiotics, glycogen, amino acids, peptides, minerals (eg, zinc, iron, copper, manganese, etc.) Etc.

  Among the preparations, for example, oral preparations can be produced as follows. Active ingredients include, for example, excipients (eg lactose, sucrose, starch, mannitol), disintegrants (eg calcium carbonate, carboxymethylcellulose calcium), or lubricants (eg talc, magnesium stearate, polyethylene glycol) Oral preparations can be made by adding and compressing, and then optionally coating by conventional methods for taste masking, enteric or persistent purposes. As the coating agent, for example, ethyl cellulose, hydroxymethyl cellulose, polyoxyethylene glycol, or the like can be used.

  In the present invention, the composition is preferably taken orally.

  If necessary, other preparations such as injections and external preparations can also be produced by conventional methods.

  In formulating, one or more medically effective active ingredients other than the active ingredient according to the present invention may be further added and blended. In administration of the active ingredient according to the present invention, one or more kinds of medically effective active ingredients other than the active ingredient according to the present invention may be administered in combination. Examples of such other active ingredients include growth hormones, calcium antagonists, protease inhibitors, steroids and the like.

  The composition according to the invention is intended not only for pharmaceutical applications but also for foods. Therefore, the food / beverage products according to the present invention comprise an effective amount of the active ingredient according to the present invention.

  As used herein, “comprising an effective amount of an active ingredient” means containing an active ingredient in such an amount that an effect as an active ingredient can be exhibited as a result of ingesting the amount of each food or drink normally consumed. . In the food and drink according to the present invention, the active ingredient according to the present invention may be blended in the food and drink as it is or in the form of the composition as described above. In addition, the food and drink according to the present invention are prepared as food and drink by adding conventional additive components such as stabilizers to the active ingredient according to the present invention, various proteins, sugars, fats, trace elements, vitamins and the like. It may be prepared by blending, liquid, semi-liquid or solid, paste, or added to a general food or drink.

  In the present invention, the “food or drink” is not a drug and is not particularly limited as long as it is ingestible by mammals, and the form thereof is any of liquid, semi-liquid or solid. May be. For this reason, the form of a drink is also included by food-drinks, for example. The food or drink may also be in the form of a dietary supplement tablet such as a supplement.

  In the present invention, the “food and beverage” includes foods classified as health foods, functional foods, foods for specified health use, dietary supplements, foods with a disease risk reduction label, or foods for the sick. The Furthermore, the term “food or drink” may be used herein to include feed when used for mammals other than humans. The food for specified health use here refers to the law in each country from the viewpoint of health when manufacturing or selling food for the purpose of preventing, improving, or alleviating the above-mentioned diseases or related conditions. Refers to food that may be subject to any of the above restrictions. Such foods can also be foods that indicate that the foods may reduce disease risk, i.e., foods with a disease risk reduction label. Here, the disease risk reduction label is a label for foods that may reduce the disease risk, and is based on or based on the standard established by the FAO / WHO Joint Food Standards Committee (Codex Committee). The display can be a defined or recognized display with reference to FIG.

  The active ingredient according to the present invention has the action as described above. For this reason, the active ingredient of the present invention is added to foods, health foods, functional foods and supplements (for example, foods containing one or more vitamins such as minerals such as calcium and magnesium and vitamin K) taken in daily life. By mix | blending, the foodstuff which has the function based on the said effect | action can be provided.

  According to the present invention, as described above, there is provided a food or drink comprising an effective amount of the active ingredient according to the present invention, which is used for suppressing or reducing the growth of leprosy in a living body.

  According to still another aspect of the present invention, there is provided a food or drink comprising an effective amount of the above active ingredient, which is used for prevention, progression suppression or improvement of leprosy or a state related thereto.

  According to another aspect of the present invention, there is provided a food / beverage product comprising an effective amount of the active ingredient according to the present invention, wherein the food / beverage product is displayed with a function of inhibiting or reducing the growth of leprosy in a living body. According to yet another aspect of the present invention, a food or drink comprising an effective amount of the active ingredient, wherein the food or drink displays a function for preventing, suppressing or improving the leprosy or a state related thereto. Is provided. Here, the function display attached to the food or drink can be, for example, any of the main body of the product, the container, the packaging, the instruction manual, the attached document, or the promotional material.

  Specific examples of the food and drink according to the present invention include juices, soft drinks, tea beverages, drinks, jelly-like beverages, functional beverages and other beverages; beer and other alcoholic beverages; rice, noodles, breads and pasta Carbohydrate-containing foods such as seafood; paste products such as fish ham, sausage and fish paste products; retort products such as curry, sauce, Chinese soup; soups; Fermented products such as yogurt, lactic acid bacteria, fermented beverages, pickles; bean products; Western confectionery such as biscuits and cookies; Japanese confectionery such as buns and sheep candy; Various kinds of confectionery; instant food such as instant soup and instant miso soup, food for microwave oven, and the like. Furthermore, health foods and drinks prepared in the form of powder, granules, tablets, capsules, liquid, paste or jelly are also included.

  In the production of foods and drinks according to the present invention, sugars, fragrances, fruit juices, food additives, stabilizers and the like that are used in normal food and drink formulation design can be added as appropriate. Manufacture of food-drinks can be implemented with reference to a manufacturing technique well-known in the said technical field. The food / beverage products according to the present invention can take various forms, and the food / beverage products according to the present invention may be produced according to known pharmaceutical production techniques. In that case, it can be produced using a carrier or formulation additive as described in the item of production of the composition according to the present invention, specifically, the carrier or production described in the column of oral preparation. Can be produced using additives. Moreover, it is good also as multifunctional food / beverage products by combining with the other component which exhibits functions other than the function in this invention, or another functional food.

  The composition according to the present invention preferably contains 0.01 to 100% by weight, more preferably 0.05 to 50% by weight, of the active ingredient relative to the total amount of the composition.

  When the composition according to the present invention is used by adding to a food or drink, the amount of the composition added to the food or drink is preferably 0.01 to 80% by mass, and 0.05 to 50% by mass. More preferably.

  When administering or ingesting the composition and food and drink according to the present invention, the dose or intake of the active ingredient according to the present invention is as follows: recipient, age and weight of recipient, symptoms, administration time, dosage form, administration method, drug It can be determined depending on the combination of. For example, when the active ingredient according to the present invention is orally administered or ingested, the daily intake amount of an adult is preferably in the range of 10 to 5000 mg as the amount of dead bacterial powder of bacteria, More preferably. These can be administered in one or several daily dosage units. These doses or intakes are calculated by calculating as necessary the dose or intake of the active ingredient per day for an adult with a body weight of 60 kg, assuming that the weight of an adult is 60 kg. be able to.

  In addition, the addition method of the microbial cell which is an active ingredient to a composition or food / beverage products has no restriction | limiting in particular, when using the dead cell body of a microbial cell, from the beginning with other raw materials used for each food / beverage product Can be added. Therefore, even if it adds to the food / beverage products which require heat processing (process under the temperature conditions which kill cells, such as heat sterilization, baking, and steaming), an effect is not impaired.

  The present invention is illustrated in detail by the following examples, but the present invention is not limited thereto.

Test method:
In the experiment, using a high sensitivity SHR / NCrj-rnu nude rats against infection of M. leprae (Mycobacterium leprae (M.leprae)). This rat can be easily obtained from related organizations including the present inventors. However, those skilled in the art can use known literature ("Leprosy in hypertensive nude rat (SHR / NCrj-run)") as necessary. , Yasuko Yogi et al., International Journal of Leprosy and Other Mycobacterial Diseases, 67: 435-445, 1999). In addition, as a component corresponding to the active ingredient of the present invention, bactericidal cell powder of EC-12 strain was used. EC-12 (trade name) (manufactured by Combi Co., Ltd.) was used as the sterilized bacterial powder by heat treatment of the EC-12 strain.

  In the experiment, 4 to 6 week-old SHR / NCrj-rnu nude rats were divided into 3 groups (2 males and 1 female) per group and divided into 2 groups, an EC-12 intake group and a non-intake group. In the lactic acid bacteria EC-12 intake group, a solid feed in which EC-12 was mixed at a rate of 0.5 mg / g into the experimental animal feed CE-2 (manufactured by Clea Japan Co., Ltd.) was prepared and freely taken every day. It was. Only the CE-2 was ingested freely in the non-ingestion group.

Seven days after the start of EC-12 intake, 2.5 × 10 ⁸ M. leprae Thai-53 strains (obtained from National Institute of Infectious Diseases, Hansen's Disease Research Center) were inoculated into the foot-pads on both legs did. After inoculation, foot-pad changes were observed over time while continuing to freely feed each feed.

  The results were as shown in FIG. 1 and FIG. The figure shows the change of foot-pad 6 months after inoculation of leprosy bacteria. Among these, FIG. 1 shows an EC-12 non-intake group, and FIG. 2 shows an EC-12 ingestion group.

As is clear from the figure, in the EC-12 non-administration group, swelling of foot-pad showing growth of leprosy bacteria was observed in all rats. On the other hand, in the EC-12 intake group, the degree of swelling was weaker than that in the EC-12 non-intake group, and almost no swelling was observed in the 2 animals in the test group.
Therefore, it can be said that these results strongly suggest that the growth of leprosy is suppressed by ingestion of EC-12.

It is a form photograph of a living body which shows the result of the Example in the case of EC-12 non-intake group (control group). It is a form photograph of a living body which shows the result of the Example in the case of EC-12 intake group.

Claims (7)

A pharmaceutical composition comprising Enterococcus faecalis as an active ingredient and used for inhibiting or reducing the growth of leprosy in a living body. The composition according to claim 1, wherein the Enterococcus faecalis is Enterococcus faecalis EC-12 (accession number FERM BP-10284).   The composition according to claim 1 or 2, wherein Enterococcus faecalis is dead cell.   The composition according to any one of claims 1 to 3, which is used for prevention, suppression or improvement of symptoms associated with leprosy or leprosy infection.   The composition according to any one of claims 1 to 4, which is orally ingested. A growth inhibitor of leprosy in the living body, or a prophylactic, progression inhibitor or ameliorant of symptoms associated with leprosy or leprosy infections, which comprises Enterococcus faecalis as an active ingredient. Enterococcus faecalis is Enterococcus faecalis EC-12 ( Enterococcus faecalis EC-12) (Accession number FERM BP-10284), The growth inhibitor of Claim 6, or a preventive agent, progress inhibitor, or improvement Agent.