JP5637716B2 - Intestinal immunity enhancer and intestinal immunity enhancement method - Google Patents
Intestinal immunity enhancer and intestinal immunity enhancement method Download PDFInfo
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Description
本発明は、有機ゲルマニウム化合物を含有する腸管免疫亢進剤及び該腸管免疫亢進剤による腸管免疫亢進方法に関する。 The present invention relates to an intestinal immunity enhancer containing an organogermanium compound and a method for enhancing intestinal immunity using the intestinal immunity enhancer.
有機ゲルマニウム化合物、とりわけ式(1)
[(Ge−CH2−CH2−COOH)2O3]n (1)
で表される有機ゲルマニウム化合物については、抗腫瘍作用(非特許文献1、2参照)、抗ウイルス作用(非特許文献3参照)、インターフェロン誘導作用NK細胞等の活性化作用のような免疫調整作用(非特許文献4参照)の他、多くの有用な生体防御作用が知られている。しかしながら、有機ゲルマニウム化合物が腸管免疫を亢進することは、知られていない。
Organic germanium compounds, especially of formula (1)
[(Ge-CH 2 -CH 2 -COOH) 2 O 3] n (1)
As for the organogermanium compound represented by the formula (1), an anti-tumor action (see Non-Patent Documents 1 and 2), an antiviral action (see Non-Patent Document 3), an interferon-inducing action, and an immunomodulating action such as an NK cell activation action. In addition to (see Non-Patent Document 4), many useful biological defense actions are known. However, it is not known that organogermanium compounds enhance intestinal immunity.
本発明の課題は、上記のような従来技術の現状を踏まえ、有機ゲルマニウム化合物を含有すると共に十分な効果を発揮する腸管免疫亢進剤、及び、該腸管免疫亢進剤による粘膜免疫方法を提供することにある。 An object of the present invention is to provide an intestinal immunity enhancer containing an organogermanium compound and exhibiting sufficient effects, and a mucosal immunization method using the intestinal immunity enhancer, based on the current state of the prior art as described above It is in.
上記課題を解決するために、本発明は以下の〔1〕〜〔5〕の態様を提供する。 In order to solve the above problems, the present invention provides the following aspects [1] to [5].
〔1〕 式(1’’’)
Ge−(CH 2 −CH 2 −COOH) 2 O 3 (1’’’)
で表される有機ゲルマニウム化合物と、乳酸菌及びオリゴ糖を含み、乳酸菌及びオリゴ糖と、有機ゲルマニウム化合物との重量比が、100:1〜5であることを特徴とする腸管免疫亢進剤。
[1] Formula (1 ''')
Ge— (CH 2 —CH 2 —COOH) 2 O 3 (1 ′ ″)
An intestinal immunity enhancer comprising an organic germanium compound represented by the formula: lactic acid bacteria and oligosaccharides, wherein the weight ratio of the lactic acid bacteria and oligosaccharides to the organic germanium compound is 100: 1 to 5.
〔2〕 乳酸菌及びオリゴ糖と、有機ゲルマニウム化合物との重量比が、100:1である〔1〕に記載の腸管免疫亢進剤。 [2] The intestinal immunity enhancing agent according to [1], wherein the weight ratio of lactic acid bacteria and oligosaccharides to the organic germanium compound is 100: 1.
〔3〕 式(1’’’)
Ge−(CH 2 −CH 2 −COOH) 2 O 3 (1’’’)
で表される有機ゲルマニウム化合物と、乳酸菌及びオリゴ糖を含む腸管免疫亢進剤を、有機ゲルマニウム化合物の投与量が、2〜5mg/Kg体重/日となるように継続的に投与することを特徴とする腸管免疫亢進方法。
[3] Formula (1 ''')
Ge— (CH 2 —CH 2 —COOH) 2 O 3 (1 ′ ″)
And an intestinal immunity enhancer comprising lactic acid bacteria and oligosaccharides, and the organic germanium compound is continuously administered so that the dose of the organic germanium compound is 2 to 5 mg / Kg body weight / day. To enhance intestinal immunity.
〔4〕 有機ゲルマニウム化合物の投与量が、2mg/Kg体重/日となるように投与する〔3〕に記載の腸管免疫亢進方法。 [4] The method for enhancing intestinal immunity according to [3], wherein the organic germanium compound is administered so that the dose thereof is 2 mg / Kg body weight / day.
〔5〕 腸管免疫亢進剤として、乳酸菌及びオリゴ糖と、有機ゲルマニウム化合物との重量比が、100:1〜5であるものを使用する〔3〕に記載の腸管免疫亢進方法。 [5] The intestinal immunity enhancing method according to [3], wherein the intestinal immunity enhancing agent is one having a weight ratio of lactic acid bacteria and oligosaccharides to the organic germanium compound of 100: 1 to 5.
本発明によれば、上記式(1)で表される有機ゲルマニウム化合物に加え、腸内環境改善に資する乳酸菌及びオリゴ糖を同時に摂取させることにより、腸管内の免疫が活性化されることを通して、生体防御機能が顕著に増強されるという効果を示す。 According to the present invention, in addition to the organogermanium compound represented by the above formula (1), by simultaneously ingesting lactic acid bacteria and oligosaccharides that contribute to improving the intestinal environment, immunity in the intestinal tract is activated, It shows the effect that the biological defense function is remarkably enhanced.
Cont:対象群
LB/OS:乳酸菌/オリゴ糖添加群
LGE:低濃度有機ゲルマニウム化合物添加群
HGE:高濃度有機ゲルマニウム化合物添加群
Cont: Target group
LB / OS: Lactic acid bacteria / Oligosaccharide addition group
LGE: Low concentration organic germanium compound addition group
HGE: High concentration organic germanium compound addition group
以下に本発明を詳細に説明する。 The present invention is described in detail below.
本発明の腸管免疫亢進剤における第1成分である有機ゲルマニウム化合物は、以下の式(1)
[(Ge−CH2−CH2−COOH)2O3]n (1)
で表されるポリ-トランス-{(2‐カルボキシエチル)-ゲルマニウムセスキオキサン}であり、この有機ゲルマニウム化合物は公知である。尚、上記式(1)は結晶状態での構造に対応するものであり、水溶液中では加水分解されて、式(1’)
(OH)3Ge−CH2−CH2−COOH (1’)
で表される構造をとる。
The organogermanium compound which is the first component in the intestinal immunity enhancing agent of the present invention has the following formula (1):
[(Ge-CH 2 -CH 2 -COOH) 2 O 3] n (1)
Poly-trans-{(2-carboxyethyl) -germanium sesquioxane}, and this organic germanium compound is known. The above formula (1) corresponds to the structure in the crystalline state and is hydrolyzed in an aqueous solution to obtain the formula (1 ′)
(OH) 3 Ge—CH 2 —CH 2 —COOH (1 ′)
The structure represented by is taken.
又、式(1’’)
Ge−(CH2−CH2−COOH)2nO3n (1’’)
や、上記従来技術に対応する非特許文献中で使用されている、式(1’’’)
Ge−(CH2−CH2−COOH)2O3 (1’’’)
も同一の化合物を示すものである。
Also, formula (1 '')
Ge- (CH 2 -CH 2 -COOH) 2n O 3n (1 '')
Or the formula (1 ′ ″) used in the non-patent literature corresponding to the above prior art.
Ge— (CH 2 —CH 2 —COOH) 2 O 3 (1 ′ ″)
Also denote the same compound.
又、本発明の腸管免疫亢進剤における第2成分である乳酸菌としては、例えば、L. fermentumやE. fecalisを挙げることができるが、本発明で使用する乳酸菌はこれらに限定されず、腸内環境改善に資するという有効性が確認される乳酸菌であればよい。 In addition, examples of the lactic acid bacteria that are the second component in the intestinal immunity enhancing agent of the present invention include L. fermentum and E. fecalis, but the lactic acid bacteria used in the present invention are not limited to these. Any lactic acid bacterium that is confirmed to be effective in improving the environment may be used.
更に、本発明の腸管免疫亢進剤における第3成分であるオリゴ糖としては、例えば、ラクチトール、乳果オリゴ糖やトレハロースを挙げることができるが、本発明で使用するオリゴ糖はこれらに限定されず、ビフィズス因子となる難消化性オリゴ糖であればよい。 Furthermore, examples of the oligosaccharide that is the third component in the intestinal immunity enhancer of the present invention include lactitol, dairy oligosaccharide, and trehalose, but the oligosaccharide used in the present invention is not limited thereto. Any indigestible oligosaccharide that becomes a bifido factor may be used.
本発明の腸管免疫亢進剤では、成分である有機ゲルマニウム化合物、乳酸菌及びオリゴ糖を広範囲の割合で含有することができるが、乳酸菌及びオリゴ糖が腸内環境の完全に資するため、有機ゲルマニウム化合物の含有量を大きく低下させることもでき、例えば乳酸菌及びオリゴ糖と、有機ゲルマニウム化合物との重量比を、100:1〜5、具体的には100:1とすることも可能である。 The intestinal immunity enhancer of the present invention can contain the components of organic germanium compounds, lactic acid bacteria and oligosaccharides in a wide range of proportions. However, since lactic acid bacteria and oligosaccharides contribute completely to the intestinal environment, The content can be greatly reduced. For example, the weight ratio of lactic acid bacteria and oligosaccharides to the organic germanium compound can be set to 100: 1 to 5, specifically 100: 1.
尚、オリゴ糖及び乳酸菌の重量比としては、例えば、100:8〜9という範囲を挙げることができる。 In addition, as a weight ratio of oligosaccharide and lactic acid bacteria, the range of 100: 8-9 can be mentioned, for example.
本発明の腸管免疫亢進剤には、その効果を失うことがない範囲で、他の成分を添加することもできる。又、適宜の剤形に製剤することも、例えば健康食品の成分として利用することもできる。 Other components can be added to the intestinal immunity enhancing agent of the present invention as long as the effect is not lost. Further, it can be formulated into an appropriate dosage form, for example, can be used as a component of health food.
一方、本発明の腸管免疫亢進方法は、上記のような本発明の腸管免疫亢進剤を継続的に投与するものである。 On the other hand, the intestinal immunity enhancing method of the present invention continuously administers the intestinal immunity enhancing agent of the present invention as described above.
本発明の腸管免疫亢進方法では、本発明の腸管免疫亢進剤の成分である乳酸菌及びオリゴ糖が腸内環境の完全に資するため、有機ゲルマニウム化合物の投与量を大きく低下させることもでき、例えば有機ゲルマニウム化合物の投与量を、2〜10mg/Kg体重/日、具体的には2mg/Kg体重/日とすることも可能である。 In the intestinal immunity enhancing method of the present invention, since the lactic acid bacteria and oligosaccharides that are the components of the intestinal immunity enhancing agent of the present invention fully contribute to the intestinal environment, the dose of the organic germanium compound can be greatly reduced. The dose of the germanium compound can be 2 to 10 mg / Kg body weight / day, specifically 2 mg / Kg body weight / day.
以下に本発明を実施例により更に詳細に説明する。 Hereinafter, the present invention will be described in more detail with reference to examples.
実施例1及び比較例1
(1)乳酸菌及びオリゴ糖成分の調製
以下の組成による乳酸菌及びオリゴ糖成分(以下、LB/OS成分と略すこともある。)を調製した。
オリゴ糖類(乳果オリゴ糖など) 71重量%
乳酸菌粉末 6重量%
ヨーグルトパウダー 6重量%
その他 17重量%
(プルラン、無水クエン酸等)
Example 1 and Comparative Example 1
(1) Preparation of lactic acid bacteria and oligosaccharide components Lactic acid bacteria and oligosaccharide components (hereinafter also abbreviated as LB / OS components) having the following composition were prepared.
Oligosaccharide (milk oligosaccharide, etc.) 71% by weight
Lactic acid bacteria powder 6% by weight
Yogurt powder 6% by weight
Other 17% by weight
(Pullan, anhydrous citric acid, etc.)
(2)マウス用飼料の調製
以下のような組成となるように、AIN93G純化飼料に基づきマウス用飼料を調製した。尚、群を示す略号の意味は以下の通りである。
Cont:対象群
LB/OS:乳酸菌/オリゴ糖添加群
LGE:低濃度有機ゲルマニウム化合物添加群
HGE:高濃度有機ゲルマニウム化合物添加群
(2) Preparation of mouse feed A mouse feed was prepared based on the AIN93G purified feed so as to have the following composition. In addition, the meaning of the symbol which shows a group is as follows.
Cont: Target group
LB / OS: Lactic acid bacteria / Oligosaccharide addition group
LGE: Low concentration organic germanium compound addition group
HGE: High concentration organic germanium compound addition group
(3)試験用マウス及び上記飼料の投与
試験用マウスは、5週齢のBALB/cCr雌マウスを採用し、一週間の予備飼育を行い順化した。予備飼育期間中は市販精製飼料D10012Mを与え、予備飼育終了後、マウスをn=6で4群に分けて、試験食を与え本飼育を行った。尚、上記LGE群(低濃度有機ゲルマニウム化合物添加群)における、LB/OS成分に対する有機ゲルマニウム化合物の重量比は約100:1で、これを有機ゲルマニウム化合物の投与量として2mg/Kg体重/日となるようにした。又、上記HGE群(高濃度有機ゲルマニウム化合物添加群)における、LB/OS成分に対する有機ゲルマニウム化合物の重量比は約100:25で、これを有機ゲルマニウム化合物の有機ゲルマニウム化合物の投与量を、これまで免疫賦活作用が確認されてきた有効摂取量である50mg/Kg体重/日となるようにした。
(3) Administration of test mouse and feed The test mouse employed a 5-week-old BALB / cCr female mouse and acclimated by pre-breeding for one week. During the preliminary breeding period, the commercially available purified feed D10012M was given, and after completion of the preliminary breeding, the mice were divided into 4 groups at n = 6, and the test diet was given for the main breeding. In the LGE group (low concentration organic germanium compound added group), the weight ratio of the organic germanium compound to the LB / OS component is about 100: 1, and this is 2 mg / Kg body weight / day as the dose of the organic germanium compound. It was made to become. In the HGE group (high concentration organic germanium compound added group), the weight ratio of the organic germanium compound to the LB / OS component is about 100: 25. The effective intake that has been confirmed to have an immunostimulatory effect was 50 mg / Kg body weight / day.
(4)結果
糞便中の分泌IgAは、腸管免疫の活性化の指標となると考えられたので、上記各群の飼料の継続摂取による腸管免疫への効果の評価を行うために分泌IgAを分析した。測定結果を図1として示す。縦軸にIgA濃度を、横軸には投与期間を示した。長期投与により、糞便中のIgA濃度は、Cont群<LB/OS群<LGE群の順で上昇する傾向にあり、特にCont群とLGE群の間には有意な差が確認された。又、LGE群と比較し、HGE群は低い値を示す傾向にあり、特に8週目では有意な差が確認された。このことから、長期継続投与への分泌IgA産生には、低濃度の有機ゲルマニウム化合物、乳酸菌及びオリゴ糖の同時摂取が有効であることが示された。
(4) Results Since secretory IgA in stool was considered to be an index of intestinal immunity activation, secretory IgA was analyzed in order to evaluate the effects on continuous intestinal immunity of each group. . The measurement results are shown in FIG. The vertical axis represents the IgA concentration, and the horizontal axis represents the administration period. After long-term administration, the fecal IgA concentration tended to increase in the order of Cont group <LB / OS group <LGE group, and a significant difference was observed between the Cont group and the LGE group. In addition, the HGE group tended to show a lower value than the LGE group, and a significant difference was confirmed particularly at the 8th week. This indicates that simultaneous intake of low-concentration organogermanium compounds, lactic acid bacteria and oligosaccharides is effective for producing secretory IgA for long-term continuous administration.
尚、図1に示したデータについては、一元配置による分散分析を実施した。異なるアルファベットは、危険率5%において有意差のあったことを示す。 In addition, about the data shown in FIG. 1, the analysis of variance by one-way arrangement was implemented. Different alphabets indicate that there was a significant difference at a risk rate of 5%.
実施例2
(1)マウス用飼料の調製
上記表1のLB/OSの組成に基づき、マウス用飼料を調製した。有機ゲルマニウム化合物の摂取量による影響評価のため、0mg/kg群、2mg/kg群、5mg/kg群及び10mg/kg群からなる4群
を設定するため、有機ゲルマニウム化合物を0%、0.0012%、0.003%、0.006%で添加して各々混合した。
Example 2
(1) Preparation of mouse feed Based on the composition of LB / OS in Table 1 above, a mouse feed was prepared. In order to evaluate the effects of intake of organic germanium compounds, 4 groups consisting of 0 mg / kg group, 2 mg / kg group, 5 mg / kg group and 10 mg / kg group were set, so that organic germanium compounds were 0%, 0.0012%, Added at 0.003% and 0.006% and mixed.
(2)試験用マウス及び上記飼料の投与
試験用マウスは、5週齢のBALB/cCr雌マウスを採用し、一週間の予備飼育を行い順化した。予備飼育期間中は市販精製飼料D10012Mを与え、予備飼育終了後、マウスをn=8で4群に分けて、上記のように調製した飼料を与え、8週間の本飼育を行った。
(2) Administration of test mouse and feed The test mouse was a 5-week-old BALB / cCr female mouse, which was acclimatized by pre-breeding for one week. During the preliminary breeding period, the commercially available purified feed D10012M was given, and after the preliminary breeding, the mice were divided into 4 groups at n = 8, and the feed prepared as described above was given, and the regular breeding was carried out for 8 weeks.
(3)結果
実施例1及び比較例1と同様に、糞便中の分泌IgAを分析した。測定結果を図2として示す。糞便中のIgA濃度は、0mg/kg群<10mg/kg群<2mg/kg群<5mg/kg群の順で上昇する傾向にあり、特に0mg/kg群と2mg/kg群の間には有意な差が確認された。いずれの濃度においても、有機ゲルマニウム化合物の低含量で混合した群では、非添加の群よりもIgA分泌量は多くなっており、このことから、乳酸菌及びオリゴ糖との同時投与には、2〜10mg/kgという低容量の有機ゲルマニウム化合物の同時摂取が有効であることが示された。
(3) Results In the same manner as in Example 1 and Comparative Example 1, secretory IgA in stool was analyzed. The measurement results are shown in FIG. Fecal IgA concentration tends to increase in the order of 0 mg / kg group <10 mg / kg group <2 mg / kg group <5 mg / kg group, especially significant between 0 mg / kg group and 2 mg / kg group The difference was confirmed. At any concentration, the group mixed with a low content of the organogermanium compound has a higher IgA secretion amount than the non-added group. Therefore, for simultaneous administration with lactic acid bacteria and oligosaccharides, 2 to Simultaneous intake of organogermanium compounds with a low volume of 10 mg / kg has been shown to be effective.
腸管免疫は、疾病の予防的側面と関連しており、上記の通りLGE群で8週目では有意な差が確認されたということは、少ない量の有機ゲルマニウム化合物、及び、乳酸菌とオリゴ糖の継続的投与が、疾病の予防的な効果を示すことを意味し、特に上記有機ゲルマニウム化合物の原料となる金属ゲルマニウムが貴重且つ高価であることを加味すれば、本発明の有用性は明らかである。 Intestinal immunity is related to the prophylactic aspect of the disease, and as described above, a significant difference was confirmed in the LGE group at the 8th week, indicating that a small amount of organogermanium compounds and lactic acid bacteria and oligosaccharides The continuous administration means that a preventive effect of the disease is exhibited, and the usefulness of the present invention is clear especially considering that the metal germanium used as the raw material of the organic germanium compound is valuable and expensive. .
尚、腸管免疫は、大野ら(実験医学、Vol.24 No.20 2006、42〜51)によれば、胸腺や抹消リンパ節等を中心とする全身免疫系とは異なる免疫系を構築しているとされる。 According to Ohno et al. (Experimental Medicine, Vol. 24 No. 20 2006, 42-51), intestinal immunity is established by constructing an immune system different from the systemic immune system centering on the thymus and peripheral lymph nodes. It is said that
Claims (2)
Ge−(CH 2 −CH 2 −COOH) 2 O 3 (1’’’)
で表される有機ゲルマニウム化合物と、乳酸菌及びオリゴ糖を含み、乳酸菌及びオリゴ糖と、有機ゲルマニウム化合物との重量比が、100:1〜5であることを特徴とする腸管免疫亢進剤。 Formula (1 ''')
Ge— (CH 2 —CH 2 —COOH) 2 O 3 (1 ′ ″)
An intestinal immunity enhancer comprising an organic germanium compound represented by the formula: lactic acid bacteria and oligosaccharides, wherein the weight ratio of the lactic acid bacteria and oligosaccharides to the organic germanium compound is 100: 1 to 5.
The intestinal immunity enhancer according to claim 1, wherein the weight ratio of lactic acid bacteria and oligosaccharides to the organic germanium compound is 100: 1.
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