JP2014196339A - Composition for prevention or treatment of anemia - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide means effective in the prevention or treatment of anemia, specifically iron deficiency anemia.SOLUTION: Disclosed is a pharmaceutical composition for enhancing the anemia prevention or treatment effects for infants in mammalian milk, the composition having as an active ingredient a bacteria belonging to the genus Bifidobacterium.

Description

  The present invention relates to a pharmaceutical composition for preventing or treating anemia and a food or drink composition for preventing or treating anemia. More specifically, the present invention relates to a pharmaceutical composition for preventing or treating anemia and a food or drink composition for preventing or treating anemia, which comprises a microorganism as an active ingredient.

  Anemia refers to a disease state in which the hemoglobin concentration in the blood unit volume has dropped to below the normal range. Symptoms associated with decreased oxygen carrying capacity such as malaise and reduced concentration occur during anemia. Furthermore, as a mechanism to compensate for anemia, that is, as a mechanism to compensate for lack of oxygen, circulatory organs such as palpitation and shortness of breath Or respiratory symptoms occur. There are various causes of anemia, but the most common type of anemia is iron deficiency anemia, which accounts for two-thirds of anemia in Japan. Iron deficiency anemia is anemia caused by a decrease in hemoglobin synthesis of erythroblasts due to iron deficiency.

  For humans, there are about 3-4 g of iron in the general adult body. Two thirds of them exist as hemoglobin-bound iron, and the remaining one-third exist in various forms such as ferritin-bound iron. In the body, most of the iron is covered by reusing iron produced by the decomposition of waste red blood cells, but about 1 mg of iron is excreted from the body per day. It is essential.

  The amount of iron absorbed from the small intestine depends on the iron content in food, the chemical structure of iron, and the iron dynamics. In addition, some foods promote iron absorption and others inhibit iron absorption. For example, it is known that black tea shows a strong iron absorption inhibitory effect, and conversely, vitamin C-containing foods such as orange juice show an iron absorption promoting effect. Meat is also an important source of heme iron, which is the most bioavailable in food. However, the iron absorption amount derived from the meal is 3 to 4 mg per day at the maximum, and the symptom cannot be improved only by diet therapy at the time of iron deficiency anemia.

  Especially in pregnant or lactating women, weanling infants, and premature babies, iron deficiency anemia is common due to the lack of adequate iron supply despite the increased demand for iron. Occurs. For example, during pregnancy, a total of about 1000 mg of iron per pregnancy is additionally required due to an increase in circulating blood volume due to pregnancy and an increase in demand for iron in the fetus and placenta as the pregnancy progresses. The Therefore, when pregnant, it is easy to get into iron deficiency anemia physiologically. If a certain level of iron deficiency anemia is observed during pregnancy, treatment with iron is administered. In addition, during the weaning period, for example, from about 4 months after birth to about 2 years of age, the demand for iron due to rapid growth and the shortage of iron in breast milk cause depletion of stored iron and iron deficiency anemia tends to occur. Therefore, iron administration is effective. In premature babies, both the amount of hemoglobin and the amount of stored iron at birth are less than those of mature babies, and iron deficiency anemia occurs more prominently earlier. There is also the idea of prophylactically administering iron preparations to pregnant women to prevent the occurrence of iron deficiency in newborns and mothers.

  As an example of the iron agent, Patent Document 1 discloses a prophylactic or therapeutic agent for anemia containing wet synthetic iron oxides.

  On the other hand, diarrhea, constipation, heartburn, upper abdominal discomfort and the like are generally known as side effects of iron preparations. In addition, administration of iron agents may inhibit the absorption of other trace metal elements such as zinc.

Under such a background, improvement of anemia using microorganisms, particularly lactic acid bacteria, has been studied. For example, there is a report that lactic acid-fermented oat straw promotes absorption of non-heme iron (Non-patent Document 1). However, on the other hand, even when lactic acid-fermented oat straw was sterilized and live lactic acid bacteria were added, there was a report that no difference was observed in iron absorption depending on the presence or absence of addition (Non-patent Document 2). ). That is, according to Non-Patent Documents 1 and 2, it is suggested that lactic acid-fermented oat straw has an iron absorption promoting effect, but lactic acid bacteria themselves do not affect iron absorption.

  Patent Document 2 discloses a composition for treating iron deficiency anemia comprising fermented milk using lactic acid bacteria belonging to Lactobacillus acidophilus and an iron salt as active ingredients. In Patent Document 2, when fermented milk is administered together with an iron-containing feed to an iron-deficient rat, the blood hemoglobin concentration is significantly higher than that when nonfat dry milk is administered together with an iron-containing feed. The effect was recognized. On the other hand, fermented milk also had an effect on body weight.

  Patent Document 3 discloses that bacteria belonging to the genus Bifidobacterium are effective in preventing or treating gastrointestinal inflammation and can be used for preventing or treating anemia. Has been. However, Patent Document 3 does not specifically disclose the effect of preventing or treating anemia.

  On the other hand, in infants, a phenomenon is known in which the hemoglobin concentration is reduced by administration of a composition containing four types of lactic acid bacteria (Non-patent Document 3). That is, it is suggested that ingestion of lactic acid bacteria may adversely affect the prevention or treatment of anemia, particularly in infants.

Japanese Patent Laid-Open No. 05-025051 JP 07-053991 A JP 2005-508617 A

Bering S et al., British Journal of Nutrition, 96: p. 80-85 (2006) Bering S, et al., British Journal of Nutrition, 98: p. 991-997 (2007) Kuitunen et al., Journal of Pediatric Gastroenterology and Nutrition, 49: p. 626-630 (2009)

  The problem to be solved by the present invention is to provide an effective means for the prevention or treatment of anemia, particularly iron deficiency anemia. The problem to be solved by the present invention is to provide a pharmaceutical composition and a food / beverage composition effective for preventing or treating iron deficiency anemia particularly in a pregnant woman or an infant.

  The present inventors have intensively studied to solve the above problems, and found that bacteria belonging to the genus Bifidobacterium show an effect of preventing or treating iron deficiency anemia, and have completed the invention.

  That is, the present invention is as follows.

  1st invention is the composition for the prevention or treatment of anemia which uses as an active ingredient the bacteria which belong to Bifidobacterium (Bifidobacterium) genus.

In a preferred embodiment of the first invention, the bacteria belonging to the genus Bifidobacterium are selected from bacteria belonging to Bifidobacterium longum and bacteria belonging to Bifidobacterium breve. The In a preferred embodiment, the bacteria belonging to Bifidobacterium longum BB536, Bifidobacterium longum ATCC 51870, Bifidobacterium longum ATCC 15707, and the strain It is selected from the mutant strains of the above strains having an effect of preventing or treating anemia that is equal to or higher than the above. In a preferred embodiment, the bacterium belonging to Bifidobacterium breve is Bifidobacterium breve M-16V, Bifidobacterium breve MCC-1274, and Bifidobacterium breve ATCC 15700, And a mutant strain of the strain having an effect of preventing or treating anemia equivalent to or higher than that of the strain.

  In a preferred embodiment of the first invention, the subject of anemia prevention or treatment is a pregnant woman or an infant.

  In a preferred embodiment of the first invention, the anemia to be prevented or treated is iron deficiency anemia.

  Moreover, the preferable aspect of 1st invention WHEREIN: The said composition is a pharmaceutical composition or a food-drinks composition. Moreover, in the preferable aspect of 1st invention, the food-drinks composition is fermented milk.

A second invention is a bacterium belonging to the genus Bifidobacterium for use in the prevention or treatment of anemia. In the second invention, preferred species and strains of bacteria belonging to the genus Bifidobacterium, preferred subjects for prevention or treatment of anemia, and preferred anemia as a subject for prevention or treatment are the same as in the first invention.
The second invention may be a composition comprising a bacterium belonging to the genus Bifidobacterium for use in the prevention or treatment of anemia. In addition, in the preferable aspect of the said composition, the said composition is a pharmaceutical composition or a food-drinks composition. Moreover, the preferable aspect of the said composition WHEREIN: A food-drinks composition is fermented milk.

  The third invention is the use of a bacterium belonging to the genus Bifidobacterium in the manufacture of a composition for preventing or treating anemia. In the third invention, preferred species and strains of bacteria belonging to the genus Bifidobacterium, preferred subjects for prevention or treatment of anemia, preferred anemia as a subject for prevention or treatment, and preferred types of compositions are the first invention. It is the same.

  The fourth invention is a composition for preventing or treating anemia comprising a food or drink containing bacteria belonging to the genus Bifidobacterium as an active ingredient. In the fourth invention, preferred species and strains of bacteria belonging to the genus Bifidobacterium, preferred subjects for the prevention or treatment of anemia, and preferred anemia as a subject for prevention or treatment are the same as in the first invention. Moreover, in the preferable aspect of 4th invention, food-drinks are fermented milk.

A fifth invention is a method for preventing or treating anemia, comprising administering a bacterium belonging to the genus Bifidobacterium to a subject in need of prevention or treatment of anemia. In the fifth invention, preferred species and strains of bacteria belonging to the genus Bifidobacterium, preferred subjects for the prevention or treatment of anemia, and preferred anemia as a subject for prevention or treatment are the same as in the first invention. In a preferred embodiment of the fifth invention, the subject in need of prevention or treatment of anemia is a mammal other than a human.

  A sixth invention is a method for producing a composition for preventing or treating anemia, comprising administering a bacterium belonging to the genus Bifidobacterium to a mammal, and using milk obtained from the mammal as a raw material A method comprising using to produce a composition for the prevention or treatment of anemia. In the sixth invention, preferred species and strains of bacteria belonging to the genus Bifidobacterium, preferred subjects for prevention or treatment of anemia, preferred anemia as a subject for prevention or treatment, and preferred types of compositions are the first invention. It is the same. In a preferred embodiment of the sixth invention, the mammal to which the bacterium belonging to the genus Bifidobacterium is administered is a mammal in pregnancy or perinatal period.

  7th invention uses the milk obtained from the mammal to which the bacteria which belong to the genus Bifidobacterium were administered as a raw material, and manufactures the composition for prevention or treatment of anemia, A method for producing a therapeutic composition. In the seventh invention, preferred species and strains of bacteria belonging to the genus Bifidobacterium, preferred subjects for the prevention or treatment of anemia, preferred anemia as a subject for prevention or treatment, and preferred types of compositions are the first invention. It is the same. In a preferred embodiment of the seventh invention, the mammal is a mammal to which a bacterium belonging to the genus Bifidobacterium is administered during pregnancy or perinatal period.

  The eighth invention is a composition for preventing or treating anemia, which is produced using milk obtained from a mammal administered with a bacterium belonging to the genus Bifidobacterium as a raw material. In the eighth invention, preferred species and strains of bacteria belonging to the genus Bifidobacterium, preferred subjects for prevention or treatment of anemia, preferred anemia as a subject for prevention or treatment, and preferred types of compositions are the first invention. It is the same. In a preferred embodiment of the eighth invention, the mammal is a mammal to which a bacterium belonging to the genus Bifidobacterium is administered during pregnancy or perinatal period.

  The ninth invention is a composition for the prevention or treatment of anemia comprising a food or drink produced using milk obtained from a mammal administered with a bacterium belonging to the genus Bifidobacterium as a raw material. In the ninth invention, preferred species and strains of bacteria belonging to the genus Bifidobacterium, preferred subjects for prevention or treatment of anemia, and preferred anemia as a subject for prevention or treatment are the same as in the first invention. Moreover, in the preferable aspect of 9th invention, food-drinks are dairy products. In a preferred embodiment of the ninth invention, the dairy product is fermented milk. In a preferred embodiment of the ninth invention, the mammal is a mammal to which a bacterium belonging to the genus Bifidobacterium is administered during pregnancy or perinatal period.

  A tenth invention is the use of milk obtained from a mammal to which a bacterium belonging to the genus Bifidobacterium is administered in the manufacture of a composition for preventing or treating anemia. In the tenth invention, preferred species and strains of bacteria belonging to the genus Bifidobacterium, preferred subjects for the prevention or treatment of anemia, preferred anemia as a subject for prevention or treatment, and preferred types of compositions are the first invention. It is the same. In a preferred embodiment of the tenth invention, the mammal is a mammal to which a bacterium belonging to the genus Bifidobacterium is administered during pregnancy or perinatal period.

The eleventh invention is milk obtained from a mammal administered with a bacterium belonging to the genus Bifidobacterium for use in the prevention or treatment of anemia. In the eleventh invention, preferred species and strains of bacteria belonging to the genus Bifidobacterium, preferred subjects for the prevention or treatment of anemia, and preferred anemia as a subject for prevention or treatment are the same as in the first invention. The first
In a preferred embodiment of one invention, the mammal is a mammal to which a bacterium belonging to the genus Bifidobacterium is administered during pregnancy or perinatal period.

  A twelfth invention is a method for preventing or treating anemia, comprising administering milk obtained from a mammal to which a bacterium belonging to the genus Bifidobacterium is administered to a subject in need of prevention or treatment of anemia, It is. In the twelfth invention, preferred species and strains of bacteria belonging to the genus Bifidobacterium, a preferred subject for prevention or treatment of anemia, and anemia preferred as a subject for prevention or treatment are the same as in the first invention. In a preferred embodiment of the twelfth invention, the mammal is a mammal to which a bacterium belonging to the genus Bifidobacterium is administered during pregnancy or perinatal period. In a preferred embodiment of the twelfth invention, the subject in need of prevention or treatment of anemia is a mammal other than a human.

The present invention includes the following embodiments.
[1] A pharmaceutical composition for enhancing the preventive or therapeutic action of anemia for infants in mammalian milk, comprising a bacterium belonging to the genus Bifidobacterium as an active ingredient.
[2] The bacterium belonging to the genus Bifidobacterium is a bacterium belonging to Bifidobacterium longum or a bacterium belonging to Bifidobacterium breve. Composition.
[3] The bacteria belonging to Bifidobacterium longum are Bifidobacterium longum ATCC BAA-999 strain, Bifidobacterium longum ATCC 51870 strain, Bifidobacterium longum ATCC 15707 strain, and The composition according to [2], which is selected from mutant strains of the strain that enhances the prevention or treatment effect of anemia equivalent to or higher than that of the strain.
[4] The bacteria belonging to the Bifidobacterium breve are Bifidobacterium breve BCCM LMG23729, Bifidobacterium breve FERM BP-11175, Bifidobacterium breve ATCC 15700, The composition according to [2], which is selected from mutant strains of the strain that enhances the prevention or treatment effect of anemia equivalent to or higher than that of the strain.
[5] The composition according to any one of [1] to [4], wherein the anemia is iron deficiency anemia.
[6] In order to enhance the preventive or therapeutic action of anemia for infants in the milk of mammals, comprising the step of preparing a bacterium belonging to the genus Bifidobacterium, which comprises the bacterium belonging to the genus Bifidobacterium as an active ingredient The manufacturing method of the pharmaceutical composition of.
[7] The method according to [6], wherein the mammal is a mammal to which a bacterium belonging to the genus Bifidobacterium is administered during pregnancy or perinatal period.
[8] The bacterium belonging to the genus Bifidobacterium is a bacterium belonging to Bifidobacterium longum or a bacterium belonging to Bifidobacterium breve [6] or [6] 7].
[9] The bacteria belonging to the Bifidobacterium longum are Bifidobacterium longum ATCC BAA-999 strain, Bifidobacterium longum ATCC 51870 strain, Bifidobacterium longum ATCC 15707 strain, and [8] The method according to [8], wherein the method is selected from mutant strains of the strain that enhances the prevention or treatment effect of anemia equivalent to or higher than that of the strain.
[10] The bacteria belonging to the Bifidobacterium breve are Bifidobacterium breve BCCM LMG23729 strain, Bifidobacterium breve FERM
The method according to [8], which is selected from the BP-11175 strain, the Bifidobacterium breve ATCC 15700 strain, and a mutant strain of the strain that enhances the anemia prevention or treatment effect equivalent to or higher than the strain.
[11] The method according to any one of [6] to [10], wherein the anemia is iron deficiency anemia.
[12] A pharmaceutical composition for preventing or treating anemia for infants, comprising milk obtained from a mammal administered with a bacterium belonging to the genus Bifidobacterium as an active ingredient.
[13] The bacterium belonging to the genus Bifidobacterium is a bacterium belonging to Bifidobacterium longum or a bacterium belonging to Bifidobacterium breve. Composition.
[14] The bacteria belonging to Bifidobacterium longum are Bifidobacterium longum ATCC BAA-999, Bifidobacterium longum ATCC 51870, Bifidobacterium longum ATCC 15707, and The composition according to [13], which is selected from mutant strains of the strain having an effect of preventing or treating anemia equivalent to or higher than that of the strain.
[15] The bacteria belonging to the Bifidobacterium breve are Bifidobacterium breve BCCM LMG23729 strain, Bifidobacterium breve FERM
The composition according to [13], which is selected from the BP-11175 strain, the Bifidobacterium breve ATCC 15700 strain, and a mutant strain of the strain having an anemia prevention or treatment effect equivalent to or higher than the strain.
[16] The composition according to any one of [12] to [15], wherein the anemia is iron deficiency anemia.
[17] A method for infants, comprising a step of producing a pharmaceutical composition for the prevention or treatment of anemia for infants using milk obtained from a mammal administered with a bacterium belonging to the genus Bifidobacterium as a raw material. A method for producing a pharmaceutical composition for preventing or treating anemia.
[18] The method according to [17], wherein the mammal is a mammal to which a bacterium belonging to the genus Bifidobacterium is administered during pregnancy or perinatal period.
[19] The bacterium belonging to the genus Bifidobacterium is a bacterium belonging to Bifidobacterium longum, or a bacterium belonging to Bifidobacterium breve [17] or [ 18].
[20] The bacteria belonging to Bifidobacterium longum are Bifidobacterium longum ATCC BAA-999 strain, Bifidobacterium longum ATCC 51870 strain, Bifidobacterium longum ATCC 15707 strain, and The method according to [19], which is selected from mutant strains of the strain having an effect of preventing or treating anemia equivalent to or higher than that of the strain.
[21] The bacteria belonging to the Bifidobacterium breve are Bifidobacterium breve BCCM LMG23729 strain, Bifidobacterium breve FERM
The method according to [19], which is selected from the BP-11175 strain, the Bifidobacterium breve ATCC 15700 strain, and a mutant strain of the strain having an anemia prevention or treatment effect equivalent to or higher than that of the strain.
[22] The method according to any one of [17] to [21], wherein the anemia is iron deficiency anemia.

The present invention improves anemia symptoms, particularly iron deficiency anemia symptoms, by administering bacteria belonging to the genus Bifidobacterium or by ingesting food and drink containing bacteria belonging to the genus Bifidobacterium. it can.
In particular, the number of red blood cells and the hemoglobin concentration can be improved by administering a bacterium belonging to the genus Bifidobacterium to a pregnant woman or by ingesting a food or drink containing a bacterium belonging to the genus Bifidobacterium.
In particular, by administering a bacterium belonging to the genus Bifidobacterium to a baby or by ingesting a food or drink containing a bacterium belonging to the genus Bifidobacterium, the red blood cell count, hemoglobin concentration, and average red blood cell hemoglobin The concentration can be improved.
Therefore, according to the present invention, it becomes possible to prevent or treat anemia, particularly iron deficiency anemia, in a pregnant woman or an infant.

FIG. 1 shows the effect of administration of bacteria belonging to the genus Bifidobacterium on red blood cell count in weanling rats. Data are shown as mean ± standard error. “*” Indicates a significant difference from the control group at a risk rate of 5% (t test). FIG. 2 shows the effect of administration of bacteria belonging to the genus Bifidobacterium on hemoglobin concentration in weanling rats. Data are shown as mean ± standard error. “*” Indicates a significant difference from the control group at a risk rate of 5% (t test). FIG. 3 shows the effect of administration of bacteria belonging to the genus Bifidobacterium on mean erythrocyte hemoglobin concentration in weanling rats. Data are shown as mean ± standard error. “*” Indicates that there is a significant difference at a risk rate of 5% (t test). FIG. 4 shows the effect of administration of bacteria belonging to the genus Bifidobacterium on the white blood cell count in weanling rats. Data are shown as mean ± standard error. FIG. 5 shows the effect of administration of bacteria belonging to the genus Bifidobacterium on body weight in weanling rats. Data are shown as mean ± standard error. FIG. 6 shows the effect of administration of various samples on the increase in hemoglobin concentration in weanling rats. Data are shown as mean ± standard deviation. "*" Indicates that there is a significant difference from the fermented milk (LA) administration group at a risk rate of 5% (Bonferroni / Dunnet). FIG. 7 shows the effect of administration of various samples on the amount of increase in red blood cell count in weanling rats. Data are shown as mean ± standard deviation. FIG. 8 shows the effect of administration of bacteria belonging to the genus Bifidobacterium on red blood cell count in pregnant rats. Data are shown as mean ± standard error. “a” and “b” indicate that there is a significant difference between different characters at a risk rate of 5% (Fisher's PLSD). FIG. 9 shows the effect of administration of bacteria belonging to the genus Bifidobacterium on hemoglobin concentration in pregnant rats. Data are shown as mean ± standard error. “a” and “b” indicate that there is a significant difference between different characters at a risk rate of 5% (Fisher's PLSD). FIG. 10 shows the effect of administration of bacteria belonging to the genus Bifidobacterium on the white blood cell count in pregnant rats. Data are shown as mean ± standard error. “a” and “b” indicate that there is a significant difference between different characters at a risk rate of 5% (Fisher's PLSD). FIG. 11 is a diagram showing an administration schedule to a maternal rat and a schedule for blood collection from a pup rat. FIG. 12 shows the effect of administration of bacteria belonging to the genus Bifidobacterium on maternal rats fed an iron-deficient diet during pregnancy and lactation on the erythrocyte count of pups. Data are shown as mean ± standard deviation. FIG. 13 shows the effect of administration of bacteria belonging to the genus Bifidobacterium on maternal rats fed with an iron-deficient diet during pregnancy and lactation on hemoglobin concentration in pups. Data are shown as mean ± standard deviation. FIG. 14 shows the effect of administration of bacteria belonging to the genus Bifidobacterium on maternal rats fed an iron-deficient diet during pregnancy and lactation on the hematocrit value of pups. Data are shown as mean ± standard deviation. FIG. 15 shows the effect of administration of bacteria belonging to the genus Bifidobacterium on maternal rats fed with an iron standard diet during pregnancy and lactation on the red blood cell count of pups. Data are shown as mean ± standard deviation. “*” Indicates that there is a significant difference from the control group at a risk rate of 1% (t test). FIG. 16 shows the effect of administration of bacteria belonging to the genus Bifidobacterium on the dam rat's hemoglobin concentration during pregnancy and lactation in maternal rats fed with an iron standard diet. Data are shown as mean ± standard deviation. “*” Indicates that there is a significant difference from the control group at a risk rate of 1% (t test). FIG. 17 shows the effect of administration of bacteria belonging to the genus Bifidobacterium on maternal rats fed with an iron standard diet during pregnancy and lactation on the hematocrit value of pups. Data are shown as mean ± standard deviation. “*” Indicates that there is a significant difference from the control group at a risk rate of 1% (t test).

  Next, an embodiment of the present invention will be described.

  The present invention relates to the prevention or treatment of anemia. Anemia includes iron deficiency anemia.

In the present invention, the prevention of anemia is to prevent the appearance of anemia symptoms in a subject. The treatment of anemia is to release the subject from the already presenting anemia symptoms.
The effect of preventing or treating anemia includes the effect of maintaining, improving, and normalizing one or more parameters of biological parameters that change depending on the degree of anemia. Examples of the biological parameter include average erythrocyte volume (MCV), average erythrocyte hemoglobin amount (MCH), and average erythrocyte hemoglobin concentration (MCHC). The mean red blood cell volume (MCV) is the average value of the volume per red blood cell, and iron deficiency anemia is suspected when the value is low. The average amount of erythrocyte hemoglobin (MCH) is the average value of the amount of hemoglobin per erythrocyte, and the value decreases during iron deficiency anemia. The mean erythrocyte hemoglobin concentration (MCHC) is an average value of the amount of hemoglobin in the erythrocyte unit volume, and the value decreases during iron deficiency anemia.
The biological parameter includes the number of red blood cells per unit volume of blood, and the value decreases during iron deficiency anemia. If both the red blood cell count and the average red blood cell hemoglobin concentration decrease, the hemoglobin concentration in the blood naturally decreases.
In addition, the biological parameter further includes the amount of stored iron. When iron is deficient, generally, a decrease in the amount of stored iron is observed before the parameter group related to red blood cells and hemoglobin decreases.
“Maintenance” refers to preventing a difference from a normal value from spreading for any of the above biological parameters. The improvement means that any of the above-mentioned biological parameters approaches a normal value. Normalization means that there is no significant difference from the normal value for any of the above biological parameters. The normal state refers to a state in which no symptoms of anemia are exhibited, particularly a state in which no symptoms of iron deficiency anemia are exhibited.

In the present invention, the composition for preventing or treating anemia contains a bacterium belonging to the genus Bifidobacterium as an active ingredient. Examples of the composition include a pharmaceutical composition and a food / beverage product composition. Bacteria belonging to the genus Bifidobacterium are preferably bacteria belonging to Bifidobacterium longum or bacteria belonging to Bifidobacterium breve.
The bacteria belonging to Bifidobacterium longum include Bifidobacterium longum BB536 (for example, ATCC BAA-999 strain), Bifidobacterium longum ATCC 51870 strain, or Bifidobacterium longum ATCC 15707 strain preferable.
Bacteria belonging to Bifidobacterium breve are preferably Bifidobacterium breve M-16V strain, Bifidobacterium breve MCC-1274 strain, or Bifidobacterium breve ATCC 15700 strain.

ATCC BAA-999 strain, ATCC 51870 strain, ATCC 15707 strain, and ATCC 15700 strain are respectively stored in the American Type Culture Collection (ATCC) (USA, Manassas University Boulevard 10801, Virginia). -Deposits with deposit numbers of 999, ATCC 51870, ATCC 15707, and ATCC 15700.
The M-16V strain is Belgian Coordinated, a Belgian conservation agency.
Deposited at the collections of Microorganisms (BCCM) (B-1000, Brussels-Ciens Street (Wetens Cup Street) 8) under the accession number BCCM LMG23729.
From August 25, 2009, the MCC-1274 strain was transferred to the National Institute of Advanced Industrial Science and Technology, Patent Biological Deposit Center (Japan 1-1, Higashi 1-1-1 Tsukuba City, Ibaraki Prefecture, Japan) from August 25, 2009. It is deposited with the deposit number of BP-11175. These strains are generally available from the above preservation agencies.

The bacterium belonging to the genus Bifidobacterium used in the present invention may be a mutant strain of the above strain as long as it has an effect of preventing or treating anemia equal to or higher than that of the above strain. Whether or not a certain mutant strain has “an equal or better effect of preventing or treating anemia” than the above strain is determined based on one or more of the above-mentioned biological parameters that vary depending on the degree of anemia, preferably the average amount of erythrocyte hemoglobin Whether or not the effect of maintaining, improving, and normalizing one or more parameters selected from (MCH) and mean erythrocyte hemoglobin concentration (MCHC) is equivalent or higher is determined by, for example, a method described in a test example described later. It can be determined by testing.
Such a mutant strain may be constructed by introducing a mutation into the strain in an artificial manner. Alternatively, mutations may be constructed by introducing mutations into the strain by treatment with a mutagen such as UV, or mutations may be constructed by introducing mutations into the strain by various gene manipulation methods. Good.

Bacteria belonging to the genus Bifidobacterium used in the present invention can be easily obtained, for example, by culturing the above strain. The culture method is not particularly limited as long as bacteria belonging to the genus Bifidobacterium can grow, and culture can be performed under appropriate conditions according to the nature of the bacteria.
For example, the culture temperature may be 25 to 50 ° C, and preferably 35 to 42 ° C. Moreover, it is preferable to perform culture | cultivation on anaerobic conditions, for example, it can culture | cultivate, ventilating anaerobic gas, such as a carbon dioxide gas. Moreover, you may culture | cultivate on microaerobic conditions, such as liquid stationary culture.

The medium for culturing bacteria belonging to the genus Bifidobacterium used in the present invention is not particularly limited, and a medium usually used for culturing bacteria belonging to the genus Bifidobacterium can be used.
That is, as the carbon source, for example, saccharides such as glucose, galactose, lactose, arabinose, mannose, sucrose, starch, starch hydrolyzate, and molasses can be used depending on utilization. As the nitrogen source, for example, ammonium salts such as ammonia, ammonium sulfate, ammonium chloride, and ammonium nitrate, and nitrates can be used. Examples of inorganic salts that can be used include sodium chloride, potassium chloride, potassium phosphate, magnesium sulfate, calcium chloride, calcium nitrate, manganese chloride, and ferrous sulfate. Organic components such as peptone, soybean powder, defatted soybean meal, meat extract, yeast extract and the like may also be used.

  As a bacterium belonging to the genus Bifidobacterium used in the present invention, the culture obtained after culturing may be used as it is, or may be used after diluting or concentrating, and using the cells recovered from the culture. May be.

The pharmaceutical composition for preventing or treating anemia of the present invention (also referred to as the pharmaceutical composition of the present invention) is not particularly limited as long as it contains bacteria belonging to the genus Bifidobacterium of the present invention as an active ingredient. It can be used as various pharmaceutical compositions containing a liquid or solid pharmaceutical carrier that is acceptable. The dosage form may be any dosage form as long as it can be taken orally, such as a suspension, powder, tablet, pill, capsule and the like. Moreover, commonly used excipients, binders, disintegrants, lubricants, coating agents, solubilizers, suspending agents, stabilizers, and other appropriate additives can be used as the pharmaceutical carrier. .

  The pharmaceutical composition for preventing or treating anemia of the present invention may or may not contain iron. As iron, for example, an iron salt is preferable. The iron salt may be either an inorganic iron salt or an organic iron salt, but is preferably a divalent iron salt from the viewpoint of absorption efficiency. When iron is contained, the content is preferably 0.2 mg / 100 g to 20000 mg / 100 g in terms of iron, and particularly preferably 2 mg / 100 g to 8000 mg / 100 g.

The content of bacteria belonging to the genus Bifidobacterium in the pharmaceutical composition for the prevention or treatment of anemia of the present invention is preferably 2 × 10 ⁷ cfu / 100 g to 2 × 10 ¹⁴ cfu / 100 g,
2 × 10 ⁸ cfu / 100 g to 2 × 10 ¹³ cfu / 100 g are particularly preferred.

  The food / beverage product composition for preventing or treating anemia of the present invention (also referred to as the food / beverage product composition of the present invention) is not particularly limited as long as it contains bacteria belonging to the genus Bifidobacterium of the present invention as an active ingredient. Examples of the product composition include dairy products, foods for adults, foods for newborns, infants, and infants.

  Examples of the dairy product include processed milk, prepared milk, powdered milk, milk beverage, fermented milk, butter and the like. As the dairy product, a dairy product for a pregnant woman or for a newborn, an infant or an infant is preferable. Examples of adult foods include liquid foods such as enteral nutrients and concentrated liquid foods, and nutritional supplements. Examples of foods for newborns, infants, and infants include formulas, powders added to mother's milk, powders added to formulas, baby foods, and the like.

Here, the formula milk is food produced by using milk or a dairy product as a main raw material and adding nutrients necessary for infants. The formula milk is mainly used as a food substitute for breast milk in the neonatal period and the infant period, or used as a food that plays a complementary role in nutrition intake. In addition, it may be used for the purpose of contributing to the intake of nutrition suitable for newborns and infants having specific congenital or acquired diseases.
Examples of formulas include infant formulas, formulas for low birth weight infants, follow-up milks, formulas for allergic diseases, lactose-free milk, and special milks for inborn errors of metabolism.

  The food / beverage product composition for preventing or treating anemia of the present invention may or may not contain iron. As iron, for example, an iron salt is preferable. The iron salt may be either an inorganic iron salt or an organic iron salt, but is preferably a divalent iron salt from the viewpoint of absorption efficiency. When iron is contained, the content is preferably 0.2 mg / 100 g to 200 mg / 100 g in terms of iron, particularly preferably 2 mg / 100 g to 80 mg / 100 g.

  The food or drink composition for preventing or treating anemia of the present invention can be produced by adding a bacterium belonging to the genus Bifidobacterium of the present invention to a general raw material of food or drink. The food / beverage product composition for preventing or treating anemia of the present invention can be produced in the same manner as in the case of producing a normal food / beverage product, except that the bacteria belonging to the genus Bifidobacterium of the present invention are added. Bacteria belonging to the genus Bifidobacterium may be added at any stage of the production process of food and drink. Moreover, food-drinks may be manufactured through the fermentation process by the bacteria which belong to the added Bifidobacterium genus.

The content of bacteria belonging to the genus Bifidobacterium in the food or beverage composition for preventing or treating anemia of the present invention is preferably 2 × 10 ⁷ cfu / 100 g to 2 × 10 ¹² cfu / 100 g, and 2 × 10 ⁸ cfu. / 100 g to 2 × 10 ¹¹ cfu / 100 g are particularly preferable.

  The food / beverage product composition for the prevention or treatment of anemia according to the present invention is a food / beverage product for which the use for preventing or treating anemia is indicated, for example, “Bifidobacterium described as“ for the prevention or treatment of anemia ”. It is preferable to sell as “food or drink containing bacteria belonging to the genus”.

  The “indication” means all acts for informing the consumer of the use, and the act of describing the use in the product or product packaging according to the food and beverage composition of the present invention, product or Transfer the product with the above-mentioned use on the product packaging, display it for delivery, transfer or delivery, display it for the purpose of importing, displaying advertisements on products, price lists or transaction documents, or distributing it Or, an act of describing the above-mentioned use in information containing these and providing it by an electromagnetic method can be exemplified.

  However, the display is preferably a display approved by the government or the like (for example, a display that is approved based on various systems determined by the government and is performed in a mode based on such approval). The indication (particularly the indication of the use of health) as the food for food (the Health Promotion Law Enforcement Regulation (April 30, 2003, Ministry of Health, Labor and Welfare Ordinance No. 86)) is most suitable.

  Note that the wording used to display the above is not limited to the wording “for the prevention or treatment of anemia”, and other words can also prevent or treat anemia. Needless to say, it can be used as long as it represents the effects that occur secondarily by the prevention or treatment of anemia.

  The administration target of the pharmaceutical composition for preventing or treating anemia and the food and beverage composition of the present invention is preferably a pregnant woman or an infant. The pregnant woman may be a pregnant woman in the perinatal period. The infant may be an infant in a weaning period. A pregnant woman is a woman in the period from the start of pregnancy to one year after delivery. Moreover, the perinatal period is a period before and after childbirth, and is a period of less than 7 days after childbirth from 22 weeks of pregnancy.

The dosage of the active ingredient in the pharmaceutical composition for preventing or treating anemia and the food / beverage product composition of the present invention is appropriately selected depending on the usage, age, sex, degree of symptoms, and other conditions. The dose of bacteria belonging to the genus Bifidobacterium as an active ingredient is preferably 2 × 10 ⁵ cfu / kg / day to 1 × 10 ¹¹ cfu / kg / day, and 2 × 10 ⁶ cfu / kg / day to 1 × 10 ¹⁰ cfu / kg / day is particularly preferred. It can be administered once a day or divided into several times. Moreover, it is good also as administration once every several days or weeks. The dose at each administration may be constant or there may be a difference.
In addition, for pregnant women, administration may be continued throughout the period from the start of pregnancy until one year after delivery, and in particular, administration may be continued throughout the perinatal period. In addition, for infants, administration may be continued throughout the infant period, and in particular, administration may be continued throughout the weaning period. Moreover, you may administer only in one part period among the said periods. Moreover, you may administer in periods other than the said period.

The pharmaceutical composition and food / beverage product composition for preventing or treating anemia of the present invention may be administered in combination with a pharmaceutical composition or food / beverage product composition containing iron. In that case, the pharmaceutical composition and food / beverage product composition for prevention or treatment of anemia of the present invention and the pharmaceutical composition or food / beverage product composition containing iron may be administered simultaneously, and are administered separately at intervals. May be. For example, an iron salt may be added to the pharmaceutical composition or the food / beverage product composition containing iron as iron. The iron salt may be either an inorganic iron salt or an organic iron salt, but is preferably a divalent iron salt from the viewpoint of absorption efficiency. Moreover, the food-drinks composition containing iron may be a food-drinks composition with high iron content, without adding iron. The amount of iron to be administered is preferably 0.002 mg / kg / day to 10 mg / kg / day, particularly preferably 0.02 mg / kg / day to 4 mg / kg / day in terms of iron.

Further, a bacterium belonging to the genus Bifidobacterium used in the present invention is administered to a mammal, or the bacterium belonging to the genus Bifidobacterium used in the present invention is ingested by a mammal, and further obtained from the mammal. By administering the given breast milk (sometimes simply referred to as milk) to the subject, anemia of the subject can be prevented or treated. The mammal to which the bacterium belonging to the genus Bifidobacterium is administered is preferably a mammal in pregnancy or perinatal period.
In addition, administering and ingesting may be collectively referred to simply as “administering” in some cases. That is, “administering” includes “taking”. The intake may be free intake, for example. Free intake means that the subject is not forced to ingest the food, but the subject is allowed to ingest the food. As an example of free intake, for example, a bacterium belonging to the genus Bifidobacterium used in the present invention can be added to food or the like and ingested by a mammal.
In addition, “administering a bacterium belonging to the genus Bifidobacterium used in the present invention to a mammal” includes, in addition to administering the bacterium itself to a mammal, the prevention or treatment of anemia of the present invention. It includes administering or ingesting a mammal-containing composition containing the bacterium, such as a pharmaceutical composition and a food and drink composition.
The dose to the mammal of the bacterium belonging to the genus Bifidobacterium used in the present invention belongs to the genus Bifidobacterium in the pharmaceutical composition and the food and beverage composition for preventing or treating anemia of the present invention described above. What is necessary is just to set suitably based on the dosage etc. of bacteria.

  The administration target of milk obtained from a mammal to which a bacterium belonging to the genus Bifidobacterium is administered is not particularly limited. For example, the milk is an administration target of the above-described pharmaceutical composition or food / beverage product composition of the present invention. Can be administered. That is, the milk can be suitably administered to, for example, a pregnant or perinatal woman, an infant, and the like, thereby preventing anemia in a pregnant or perinatal woman, an infant, and the like. Can improve symptoms. The dose of the milk is not particularly limited, and can be, for example, an amount that a woman or an infant in pregnancy or perinatal normally ingests milk.

  The milk is preferably human breast milk, and particularly preferably milk from mammals other than humans. Examples of mammals other than humans include mammals generally used in dairy farming. Examples of such mammals include cows, goats, sheep, horses, and buffaloes, with cattle being preferred. Milk can be obtained from a mammal, for example, by milking. The milk may be obtained from the mammal during the period when the bacteria belonging to the genus Bifidobacterium are administered to the mammal, and the mammal after the period when the bacteria belonging to the genus Bifidobacterium are administered to the mammal. May be obtained from

Further, a bacterium belonging to the genus Bifidobacterium used in the present invention is administered to a mammal, or a bacterium belonging to the genus Bifidobacterium used in the present invention is ingested by a mammal, and obtained from the mammal. By using milk as a raw material, a composition for preventing or treating anemia can be produced. The mammal to which the bacterium belonging to the genus Bifidobacterium is administered is preferably a mammal in pregnancy or perinatal period.
The said composition can be manufactured as a pharmaceutical composition or a food-drinks composition. Moreover, dairy products can be illustrated as a food-drinks composition. The food / beverage composition is produced in the same manner as in the production of normal food / beverage products, except that milk obtained from a mammal to which a bacterium belonging to the genus Bifidobacterium used in the present invention is administered is used as a raw material. it can. The food / beverage composition may be milk itself.
By administering the composition to an application subject, anemia of the subject can be prevented or treated. The administration subject in particular of the said composition is not restrict | limited, For example, the said composition can be administered to the administration subject of the pharmaceutical composition or food-drinks composition of this invention mentioned above. That is, the milk can be suitably administered to, for example, a pregnant or perinatal woman, an infant, and the like, thereby preventing anemia in a pregnant or perinatal woman, an infant, and the like. Can improve symptoms. The dose of the composition is not particularly limited, and can be, for example, an amount that a woman or an infant in pregnancy or perinatal period normally takes dairy products.

  Next, the present invention will be described in detail by test examples.

Test Example 1: Effect of Bifidobacterium Sample Administration in Weaning Period According to the present invention, the effect of Bifidobacterium sample administration was tested on weanling rats.

<Preparation of Bifidobacterium sample>
Bifidobacterium breve M-16V strain was used as a bacterium belonging to the genus Bifidobacterium. 48 mg of Bifidobacterium breve M-16V strain powder (2.1 × 10 ¹¹ cfu / g, Morinaga Milk Industry Co., Ltd.) powdered in starch is suspended in phosphate buffered saline (PBS). / 400 μl (1 × 10 ¹⁰ cfu / 400 μl) to prepare a Bifidobacterium sample.

<Bifidobacterium sample administration in weaning period>
As a maternal rat, F344 / N strain pregnant rats were purchased from Japan SLC. Maternal rats were fed MR STOCK as food and allowed to freely ingest. The pups born to the mother rats were weaned at 21 days after birth, and divided into two groups so that there was no bias among the groups according to body weight, parent, and sex. It shows in Table 1 about each group.

For 14 days from day 21 to day 34, pups were fed MR STOCK as a feed and allowed to freely eat. Further, the above-mentioned Bifidobacterium sample was administered once a day at 1 × 10 ¹⁰ cfu / rat / day to the test group, and the same amount of starch (48 mg / rat / day) was administered to the control group a day. A single dose was administered. The amount of iron in MR STOCK is 20.2 mg / 100 g, and the amount of iron in feed AIN-93G used as a standard for nutritional studies of rats including the breeding season is 4.01 mg / 100 g. MR STOCK contained about 5 times as much iron as the standard feed AIN-93G, and the amount of iron in MR STOCK was sufficient.

  Dissection was performed at the age of 35 days, and blood was collected from the inferior vena cava. The obtained blood sample was analyzed with a KX-21NV hemocytometer (manufactured by Sysmex Corporation). Statistical analysis was performed by t-test.

The red blood cell count is shown in FIG. As a result, the test group showed a significantly higher red blood cell count with a risk rate of 5% than the control group.
The hemoglobin concentration is shown in FIG. As a result, the test group showed a significantly higher hemoglobin concentration at a risk rate of 5% than the control group.
The average erythrocyte hemoglobin concentration is shown in FIG. As a result, the test group showed a significantly higher average erythrocyte hemoglobin concentration at a risk rate of 5% than the control group.

  On the other hand, there was no significant difference in the number of leukocytes depending on the presence or absence of administration of the Bifidobacterium sample (FIG. 4). In addition, there was no significant difference in body weight depending on whether or not the Bifidobacterium sample was administered (FIG. 5).

  As described above, improvement of biological parameters related to anemia was observed by administration of the Bifidobacterium sample. Therefore, the prevention or treatment effect of anemia by administration of bacteria belonging to the genus Bifidobacterium was observed in weanling rats. Here, while the number of red blood cells, the hemoglobin concentration, and the average red blood cell hemoglobin concentration related to the amount of iron in the living body were improved, no significant differences were observed in the body weight and the number of white blood cells not related to the amount of iron.

Test Example 2: Comparison of recovery effect from iron deficiency anemia in weaning period Next, according to the present invention, Bifidobacterium bacterial powder sample, fermented milk sample, and Bifidobacterium-containing fermented milk according to the present invention A comparative test was conducted on the recovery effect from iron deficiency anemia when each of the samples was administered.

<Preparation of Bifidobacterium sample>
Bifidobacterium breve M-16V strain and Bifidobacterium longum BB536 strain were used as bacteria belonging to the genus Bifidobacterium. Bifidobacterium breve M-16V strain powder (2.1 × 10 ¹¹ cfu / g, Morinaga Milk Industry Co., Ltd.) and Bifidobacterium longum BB536 strain powder (1 0.0 × 10 ¹¹ cfu / g (manufactured by Morinaga Milk Industry Co., Ltd.) was suspended in phosphate buffered saline (PBS), adjusted to 2.5 × 10 ⁹ cfu / 400 μl, and Bifidobacterium sample (M-16V
Bacterial powder, BB536 bacterial powder).

<Preparation of fermented milk sample>
A raw material was prepared by adding 0.3% by weight of yeast extract to skim milk having a solid content of nonfat milk of 11% by weight. This raw material is sterilized at 90 ° C. for 10 minutes, cooled to 38 ° C., and added with about 3% by weight of Lactobacillus acidophilus LAC-361 strain as a lactic acid bacteria starter, and fermented at 37 ° C. for 18 hours. Fermented milk (LA) was prepared.
In addition, fermented milk (BB536) was prepared by adding cell powder of Bifidobacterium longum BB536 strain to fermented milk (LA) so as to be 2.5 × 10 ⁹ cfu / 2 ml.

<Various sample administration in weaning period>
As a maternal rat, F344 / N strain pregnant rats were purchased from Japan SLC. Maternal rats were fed with AIN-93G (Oriental Yeast Co., Ltd.) as a feed and allowed to eat freely.

  A pup rat born from a mother animal rat was weaned at 21 days after birth. For 14 days from age 21 to day 34, pups were fed with AIN-93G-based diet (iron-deficient diet) deficient in iron as a diet to induce iron-deficiency anemia. Blood was collected from the tail vein at the age of 35 and confirmed to be anemic, and the groups were divided so that there was no bias among the groups according to hemoglobin concentration, body weight, and sex.

Next, the feed is changed to AIN-93G (iron standard diet), which is a standard diet, and raised for 7 days from age 35 to age 41, and recovery from iron deficiency anemia induced above is recovered. planned. Meanwhile, in addition to AIN-93G, each group was administered the above fermented milk sample at 2 ml / rat / day or the Bifidobacterium sample once a day at 2.5 × 10 ⁹ cfu / rat / day. .
Table 2 shows the administration sample, the number of specimens, and the dosage for each group. The iron content of the iron-deficient diet and the standard diet were 0.2 mg / 100 g and 4.01 mg / 100 g, respectively. AIN-93G is a feed that is used as a standard for nutritional studies on rats including the breeding season.

  Dissection was performed at the age of 42 days, and blood was collected from the inferior vena cava. The obtained blood sample was analyzed with a KX-21NV hemocytometer (manufactured by Sysmex Corporation). From the obtained blood data, the amount of increase in hemoglobin concentration and red blood cell count from age 35 to day 42 was determined, and statistical analysis was performed by Bonferroni / Dunnet.

The amount of increase in hemoglobin concentration is shown in FIG. As a result, each of the administration groups of M-16V bacterial powder, BB536 bacterial powder, and fermented milk (BB536) showed significantly higher hemoglobin concentration with a risk rate of 5% than the fermented milk (LA) administration group. It was.
Moreover, it shows in FIG. 7 about the increase amount of red blood cell count. As a result, it was recognized that each of the administration groups of M-16V bacterial powder, BB536 bacterial powder, and fermented milk (BB536) had a higher increase in the number of red blood cells than the fermented milk (LA) administration group.

  On the other hand, there was no significant difference in body weight between the administration groups of each sample.

As mentioned above, compared with the case where fermented milk (LA) is administered by administration of each of M-16V bacterial powder, BB536 bacterial powder and fermented milk (BB536), the degree of improvement of biological parameters related to anemia is increased. Improved.
Therefore, the prevention or treatment effect of anemia by administration of bacteria belonging to the genus Bifidobacterium was observed in weanling rats. Moreover, the effect equivalent to the case where M-16V bacterial powder or BB536 bacterial powder is administered was also observed by adding BB536 bacterial powder to fermented milk.

Test Example 3: Effect of Bifidobacterium Sample Administration in Pregnancy and Lactation Next, according to the present invention, the effect of Bifidobacterium sample administration was tested on gestation and lactation rats.

<Preparation of Bifidobacterium sample>
Bifidobacterium longum BB536 strain was used as a bacterium belonging to the genus Bifidobacterium. Bifidobacterium longum BB536 strain powder triturated in starch (1.1 × 10 ¹¹ cfu / g, Morinaga Milk Industry Co., Ltd.) was suspended in PBS and 9 mg / ml (1 × 10 ⁹ cfu / ml). ) To prepare a Bifidobacterium sample.

<Bifidobacterium sample administration during pregnancy and lactation>
Brown Norway strain pregnant and non-pregnant rats were purchased from Charles River Japan. Pregnant rats were divided into two groups, a control group and a test group. In addition, non-pregnant rats were designated as non-pregnant groups. It shows in Table 3 about each group.

For 12 days from the 14th day of pregnancy to the 4th day after childbirth, F-2 was fed as a feed and allowed to freely ingest. In addition, non-pregnant rats were similarly fed with F-2 and allowed to freely take them. Further, the above-mentioned Bifidobacterium sample was administered once a day at 1 × 10 ⁹ cfu / rat / day to the test group, and the same amount of PBS (1 ml / rat / day) was administered to the control group once a day. Administered once. In the non-pregnant group, neither Bifidobacterium sample nor PBS was administered.
Since the amount of iron contained in F-2 is 30 mg / 100 g and the amount of iron contained in standard feed AIN-93G is 4.01 mg / 100 g, F-2 contains standard feed AIN-93G. About 7.5 times the amount of iron was contained, and the amount of iron in F-2 was sufficient.

  Dissection was performed on day 5 after delivery, and blood was collected from the inferior vena cava. The obtained blood sample was analyzed by Cell Tac MEK-5254 blood cell counter (manufactured by Nihon Kohden Co., Ltd.). As a statistical analysis, 1-way ANOVA was performed, and multiple comparison test by Fisher's PLSD was further performed.

The red blood cell count is shown in FIG. As a result, the control group showed significantly lower red blood cell count than the non-pregnant group. On the other hand, with the administration of the Bifidobacterium sample, the test group showed a significantly higher red blood cell count with a 5% risk than the control group, and the red blood cell count was improved to the extent that no significant difference was observed with the non-pregnant group. It was done.
The hemoglobin concentration is shown in FIG. As a result, the control group showed a significantly lower hemoglobin concentration than the non-pregnant group. On the other hand, the administration of the Bifidobacterium sample showed a significantly higher hemoglobin concentration at a risk rate of 5% compared to the control group, and the hemoglobin concentration was improved to the extent that no significant difference was observed with the non-pregnant group. It was done.

  On the other hand, there was no significant difference in the number of leukocytes depending on whether or not the Bifidobacterium sample was administered (FIG. 10).

As described above, the administration of the Bifidobacterium sample improved the biological parameters related to anemia, and was able to restore the red blood cell count and hemoglobin concentration to the same level as when non-pregnant.
Therefore, in pregnancy and lactation rats, the effect of preventing or treating anemia was confirmed by administration of bacteria belonging to the genus Bifidobacterium. Here, while the number of red blood cells and the hemoglobin concentration related to the amount of iron in the living body were improved, no significant difference was observed in the number of white blood cells not related to the amount of iron.

Test Example 4: Anemia-preventing effect of bifidobacterium sample administration to newborn rats during pregnancy and lactation period Next, according to the present invention, a bifidobacterium sample is administered to pregnant mother rats The newborn rat was then tested for the anemia-preventing effect in the newborn rat when it was fed from a maternal rat to which the Bifidobacterium sample had been administered, even in the lactation stage.

<Preparation of Bifidobacterium sample>
Bifidobacterium longum BB536 strain was used as a bacterium belonging to the genus Bifidobacterium. Bifidobacterium longum BB536 strain powder doubled in starch (1.0 × 10 ¹¹ cfu / g, Morinaga Milk Industry Co., Ltd.) was suspended in PBS and adjusted to 2.5 × 10 ⁹ cfu / 400 μl. A Bifidobacterium sample was used.

<Bifidobacterium sample administration during pregnancy and lactation>
F344 / N strain pregnant rats were purchased from Japan SLC. Pregnant rats were divided into 4 groups, which were iron deficient diet (control group and test group) and iron standard diet (control group and test group). It shows in Table 4 about each group.

AIN-93G-based diet (iron-deficient diet) in which the mother rat was deprived of iron as a diet for a total of 21 days from the 7th to 14th day of pregnancy and thereafter until the 7th day, or AIN- 93G (iron standard diet) was fed and freely ingested. Further, the above-mentioned Bifidobacterium sample was administered once a day at 2.5 × 10 ⁹ cfu / rat / day to the mother rats of the test group, and the same amount of starch (25 mg / rat / day) was administered to the control group. Day) was administered once a day.

  At 7 days after birth, the rat pups were dissected and blood was collected from the inferior vena cava. The obtained blood sample was analyzed with a blood cell counter (manufactured by Sysmex Corporation, model: KX-21NV). Statistical analysis was performed by t-test.

  In addition, the administration schedule to a mother animal rat and the schedule to blood collection from a born pup rat are shown in FIG.

In the iron-deficient diet group, the erythrocyte count of the pups (blood collection: control group 13, test group 13), the hemoglobin concentration of the pups (blood collection: control group 13, test group 13), and the hematocrit value of the pups (blood collection) Numbers: control group 13, test group 13) are shown in FIGS. 12, 13 and 14 respectively.
As a result, the test group showed a tendency that the number of red blood cells (FIG. 12), hemoglobin concentration (FIG. 13), and hematocrit value (FIG. 14) increased as compared with the control group.

In the iron standard diet group, the number of erythrocytes of the pups (blood collection: control group 21, test group 19), hemoglobin concentration of pups (blood collection: control group 21, test group 19), and hematocrit value of pups (blood collection) Numbers: control group 21 and test group 19) are shown in FIGS. 15, 16 and 17, respectively.
As a result, the test group showed a significantly higher red blood cell count with a risk rate of 1% than the control group (FIG. 15). In addition, the test group showed a significantly higher hemoglobin concentration with a risk rate of 1% than the control group (FIG. 16). Moreover, the test group showed a significantly high hematocrit value at a risk rate of 1% with respect to the control group (FIG. 17).

As described above, in pups born from a group of dams fed an iron-deficient diet, administration of the Bifidobacterium sample to the gestation and lactation dams compared to the control group to which starch was administered. Thus, an improvement tendency of the red blood cell count, hemoglobin concentration, and hematocrit value was observed.
On the other hand, in pups born from a group of dams fed the iron standard diet, administration of the Bifidobacterium sample to the gestation and lactation dams compared to the control group to which starch was administered. A significant improvement in the above-mentioned anemia-related biological parameters was observed.
Therefore, administration of a Bifidobacterium sample to a maternal rat during pregnancy and lactation showed an anemia-preventing effect in newborn rats.

  In this test example, the newborn rat was bred by suckling from the mother animal rat. Therefore, it is considered that the breast milk of a maternal rat administered with the Bifidobacterium sample has an effect of preventing anemia of a pup rat to be sucked.

  EXAMPLES The present invention will be described in more detail below with reference to examples, but the scope of the present invention is not limited to the following examples.

Example 1: Prepared milk powder Desalinated milk whey protein powder (New Zealand Daily Board) 10 kg, Milk casein powder (New Zealand Daily Board) 6 kg, Lactose (Mirai) 48 kg, Mineral mixture (Tonda Pharmaceutical) 920 g (including 93 g of iron) and 32 g of vitamin mixture (Tanabe Seiyaku Co., Ltd.) are dissolved in 300 kg of hot water, heated and dissolved at 90 ° C. for 10 minutes, and 28 kg of prepared fat (manufactured by Taiyo Oil & Fats) is added. And homogenized. Thereafter, sterilization and concentration steps were performed and spray-dried to prepare about 93 kg of prepared milk powder. To this, 100 g of Bifidobacterium breve M-16V strain powder (1.8 × 10 ¹¹ cfu / g, Morinaga Milk Industry Co., Ltd.) triturated in starch was added as bifidobacteria to prepare a bifidobacteria mixture. About 93 kg of milk powder was prepared. When the prepared powdered milk was dissolved in water to prepare a milk preparation with a standard milk concentration of 14% (w / V), the iron content in the milk preparation was 1.4 mg / 100 ml. The number of bifidobacteria was 2.7 × 10 ⁹ cfu / 100 ml. This formula can be used for the prevention or treatment of anemia.

Example 2: Powdered milk Skim milk powder (manufactured by Morinaga Milk Industry Co., Ltd.) 26.3 kg, corn starch (manufactured by Matsutani Chemical Co., Ltd.) 9 kg, dextrin (manufactured by Matsutani Chemical Co., Ltd.) 51.3 kg, mineral mixture (manufactured by Merck) 7.5 kg ( 17.6 g as iron) and 400 g of vitamin mixture (DSM) are dissolved in 450 kg of warm water, heated and dissolved at 90 ° C. for 10 minutes, and 5.5 kg of prepared fat (manufactured by Taiyo Oil & Fats) is added Homogenized. Thereafter, sterilization and concentration steps were performed and spray-dried to prepare about 100 kg of milk powder. To this, 200 g of Bifidobacterium longum BB536 strain powder (1 × 10 ¹¹ cfu / g, Morinaga Milk Industry Co., Ltd.) triturated in starch was added as bifidobacteria to prepare about 100 kg of bifidobacteria mixed milk powder. did. When the obtained milk powder was dissolved in water to prepare a milk preparation with a total solid content of 13% (w / V), the iron content in the milk preparation was 2.3 mg / 100 ml, and the number of bifidobacteria was 2.6. × 10 ⁹ cfu / 100 ml. This milk powder can be used for the prevention or treatment of anemia.

Example 3: Liquid food (powder)
Whey protein enzyme degradation product (Morinaga Milk Co., Ltd.) 10.8 kg, dextrin (Showa Sangyo Co., Ltd.) 36 kg, and a small amount of water-soluble vitamins and minerals (including 2.4 g of iron) are dissolved in 200 kg of water and the aqueous phase is dissolved. Was prepared in a tank. Separately, soybean salad oil (manufactured by Taiyo Yushi Co., Ltd.) 3 kg, palm oil (manufactured by Taiyo Yushi Co., Ltd.) 8.5 kg, safflower oil (manufactured by Taiyo Yufu Co., Ltd.) 2.5 kg, lecithin (manufactured by Ajinomoto Co., Inc.) 0.2 kg, An oil phase was prepared by mixing and dissolving 0.2 kg of a fatty acid monoglyceride (manufactured by Kao Corporation) and a small amount of a fat-soluble vitamin. The oil phase was added to the aqueous phase in the tank, stirred and mixed, then heated to 70 ° C. and further homogenized with a homogenizer at a pressure of 14.7 MPa. Then, after sterilizing at 90 ° C. for 10 minutes, it was concentrated and spray-dried to prepare about 59 kg of intermediate product powder. 50 kg of this intermediate product powder, 6.8 kg of sucrose (manufactured by Hokuren Co., Ltd.), 167 g of amino acid mixed powder (manufactured by Ajinomoto Co., Inc.), and Bifidobacterium longum BB536 strain powder dispersed in starch as bifidobacteria 110 g (1 × 10 ¹¹ cfu / g, manufactured by Morinaga Milk Industry Co., Ltd.) was added and mixed uniformly to produce about 57 kg of enteral nutrition food powder. The iron content in the powder was 3.7 mg / 100 g, and the number of bifidobacteria was 1.9 × 10 ¹⁰ cfu / 100 g. This liquid food can be used for the prevention or treatment of anemia.

Example 4: Capsules Lactose (Wako Pure Chemical Industries) 400g, Corn starch (Nisshin Flour Mills) 160g, Crystalline cellulose (Wako Pure Chemical Industries) 200g, Mineral mixture (Tonda Pharmaceutical) 40g (Iron) 4 g), lactoferrin (manufactured by Morinaga Milk Co., Ltd.), 600 g, and Bifidobacterium longum BB536 strain powder dispersed in starch as bifidobacteria (1 × 10 ¹¹ cfu / g, Morinaga Dairy Co., Ltd.) 600 g) were screened through 50 mesh screens and mixed uniformly. Using a fully automatic capsule filling machine (manufactured by Cese Pedini), the powder was filled into capsules (manufactured by Elanco Japan, No. 1 gelatin capsule, Op. Yellow No. 6 Body, empty weight 75 mg) with an inner volume of 275 mg, and iron 0 7000 mg capsules containing 0.55 mg and 82 mg (8 × 10 ⁹ cfu) of Bifidobacterium powder were prepared. This capsule can be used for the prevention or treatment of anemia.

  The bacterium belonging to the genus Bifidobacterium of the present invention is effective in preventing or treating iron deficiency anemia. Therefore, by including bacteria belonging to the genus Bifidobacterium in a pharmaceutical composition or a food and beverage composition, a pharmaceutical composition for preventing or treating iron deficiency anemia, particularly iron deficiency anemia in pregnant women or infants, and food and drink Product compositions can be provided.

Claims (22)

  A pharmaceutical composition for enhancing the preventive or therapeutic action of anemia for infants in mammalian milk, comprising a bacterium belonging to the genus Bifidobacterium as an active ingredient.   The composition according to claim 1, wherein the bacterium belonging to the genus Bifidobacterium is a bacterium belonging to Bifidobacterium longum or a bacterium belonging to Bifidobacterium breve. .   Bacteria belonging to Bifidobacterium longum enhance Bifidobacterium longum ATCC BAA-999 strain, Bifidobacterium longum ATCC 15707 strain, and the prevention or treatment of anemia equivalent to or higher than the strain The composition according to claim 2, which is selected from mutant strains of the strain.   Bifidobacterium breve bacteria belonging to Bifidobacterium breve BCCM LMG23729 strain, Bifidobacterium breve FERM BP-11175 strain, Bifidobacterium breve ATCC 15700 strain, and equivalent to the above strain The composition according to claim 2, which is selected from the mutant strains of the strain that enhances the above-described anemia prevention or treatment action.   The composition according to any one of claims 1 to 4, wherein the anemia is iron deficiency anemia.   A pharmaceutical composition for enhancing the preventive or therapeutic action of anemia for infants in mammal milk, comprising the step of formulating a bacterium belonging to the genus Bifidobacterium, comprising a bacterium belonging to the genus Bifidobacterium as an active ingredient Manufacturing method.   The method according to claim 6, wherein the mammal is a mammal to which a bacterium belonging to the genus Bifidobacterium is administered during pregnancy or perinatal period.   The bacterium belonging to the genus Bifidobacterium is a bacterium belonging to Bifidobacterium longum or a bacterium belonging to Bifidobacterium breve. Method.   Bacteria belonging to Bifidobacterium longum enhance Bifidobacterium longum ATCC BAA-999 strain, Bifidobacterium longum ATCC 15707 strain, and the prevention or treatment of anemia equivalent to or higher than the strain The method according to claim 8, which is selected from a mutant strain of the strain.   Bifidobacterium breve bacteria belonging to Bifidobacterium breve BCCM LMG23729 strain, Bifidobacterium breve FERM BP-11175 strain, Bifidobacterium breve ATCC 15700 strain, and equivalent to the above strain The method according to claim 8, which is selected from the mutant strains of the strain that enhance the preventive or therapeutic action of the above anemia.   The method according to any one of claims 6 to 10, wherein the anemia is iron deficiency anemia.   A pharmaceutical composition for the prevention or treatment of anemia for infants, comprising milk obtained from a mammal administered with a bacterium belonging to the genus Bifidobacterium as an active ingredient.   The composition according to claim 12, wherein the bacterium belonging to the genus Bifidobacterium is a bacterium belonging to Bifidobacterium longum or a bacterium belonging to Bifidobacterium breve. .   Bacteria belonging to Bifidobacterium longum have Bifidobacterium longum ATCC BAA-999 strain, Bifidobacterium longum ATCC 15707 strain, and anemia prevention or treatment effect equivalent to or higher than that strain The composition according to claim 13, which is selected from mutant strains of the strain.   Bifidobacterium breve bacteria belonging to Bifidobacterium breve BCCM LMG23729 strain, Bifidobacterium breve FERM BP-11175 strain, Bifidobacterium breve ATCC 15700 strain, and equivalent to the above strain The composition according to claim 13, which is selected from the mutant strains of the strain having the above anemia prevention or treatment effect.   The composition according to any one of claims 12 to 15, wherein the anemia is iron deficiency anemia.   Infants comprising a step of obtaining milk from a mammal to which a bacterium belonging to the genus Bifidobacterium is administered, and a step of producing a pharmaceutical composition for preventing or treating anemia for infants using the milk as a raw material For producing a pharmaceutical composition for the prevention or treatment of anemia.   The method according to claim 17, wherein the mammal is a mammal to which a bacterium belonging to the genus Bifidobacterium is administered during pregnancy or perinatal period.   The bacterium belonging to the genus Bifidobacterium is a bacterium belonging to Bifidobacterium longum or a bacterium belonging to Bifidobacterium breve. Method.   Bacteria belonging to Bifidobacterium longum have Bifidobacterium longum ATCC BAA-999 strain, Bifidobacterium longum ATCC 15707 strain, and anemia prevention or treatment effect equivalent to or higher than that strain 20. A method according to claim 19, selected from a mutant strain of said strain.   Bifidobacterium breve bacteria belonging to Bifidobacterium breve BCCM LMG23729 strain, Bifidobacterium breve FERM BP-11175 strain, Bifidobacterium breve ATCC 15700 strain, and equivalent to the above strain The method according to claim 19, wherein the method is selected from mutant strains of the strain having the above-described anemia prevention or treatment effect.   The method according to any one of claims 17 to 21, wherein the anemia is iron deficiency anemia.

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