TW201427603A - Allergy improving agent - Google Patents

Allergy improving agent Download PDF

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TW201427603A
TW201427603A TW102142097A TW102142097A TW201427603A TW 201427603 A TW201427603 A TW 201427603A TW 102142097 A TW102142097 A TW 102142097A TW 102142097 A TW102142097 A TW 102142097A TW 201427603 A TW201427603 A TW 201427603A
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galactooligosaccharide
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Hiroaki Matsuyama
Satoshi Higurashi
Ken Katoh
Seiichiro Aoe
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Megmilk Snow Brand Co Ltd
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Abstract

An object of the invention is to provide an allergy improving agent which has the effect of alleviating disruption to the immune regulatory system in vivo by suppressing the secretion of inflammatory cytokines, and to provide allergy improving products such as foods, beverages and medicines that contain this allergy improving agent. The allergy improving agent of the invention contains, as an active ingredient, a galacto-oligosaccharide having a structural formula represented by O- β -D-galactopyranosyl-(1 to 6)-[O- β -D-galactopyranosyl-(1 to 4)]-D-glucose or O- β -D-galactopyranosyl-(1 to 4)-[O- β -D-galactopyranosyl-(1 to 6)]-D-glucose.

Description

過敏症改善劑 Allergy improver

本發明係關於過敏症改善劑。更詳而言之,本發明係關於一種過敏症改善劑,其於過敏症改善作用優秀,藉由抑制代謝性之發炎並抑制發炎性之細胞激素之分泌以改善生物體內之免疫調節系統混亂。更者,本發明係關於含有過敏症改善劑之過敏症改善用飲食品、過敏症改善用營養組成物、過敏症改善用飼料或過敏症改善用醫藥品。 The present invention relates to an allergy improving agent. More specifically, the present invention relates to an allergy-improving agent which is excellent in allergy-improving effects and which improves immune system disorder in a living body by inhibiting metabolic inflammation and inhibiting secretion of inflammatory cytokines. Furthermore, the present invention relates to a food or drink for allergy-improving food, a nutritional composition for allergy improvement, a food for allergy improvement, and a medicine for improving allergy.

近年,在包括日本之先進國家中,異位性皮膚炎或花粉症等之各種過敏症疾病增加中,成為很大的社會問題。關於以異位性皮膚炎為首之過敏症疾病的發作,涉及遺傳性的因素與環境因素兩者,關於近年的過敏症疾病患者數的增加,推測為受大氣污染或生活方式的變化,飲食生活的變化等之環境因素的影響很大。又,過敏症疾病的發生機制係為複雜,仍舊也有許多尚未清楚的部份,但漸漸地知曉其原因在於免疫調節系統的混亂。輔助T細胞在免疫調節中進行重要之作用,在該輔助T細胞具有輔助T1(以下稱為「Th1」)細胞與輔助T2(以下成為「Th2」)細胞2種,分別透過細胞激素的產生而控制細胞性免疫與體液性免疫。Th1細胞與Th2細胞,藉由各自生產之細胞激素抑制彼此的活化,而保持平衡(Th1/Th2平衡),但因某些原因導致Th2細胞的誘導增強則使過敏症疾病出現症狀。 In recent years, in various advanced countries including Japan, a variety of allergic diseases such as atopic dermatitis or hay fever have become a major social problem. Regarding the onset of allergic diseases including atopic dermatitis, both genetic factors and environmental factors are involved. The increase in the number of allergic diseases in recent years is presumed to be caused by air pollution or lifestyle changes. Environmental factors such as changes have a great impact. Moreover, the mechanism of allergic diseases is complicated, and there are still many unclear parts, but it is gradually known that the cause is the confusion of the immune regulation system. The helper T cells play an important role in immune regulation, and the helper T cells have two types of helper T1 (hereinafter referred to as "Th1") cells and helper T2 (hereinafter referred to as "Th2") cells, respectively, through the production of cytokines. Control cellular immunity and humoral immunity. Th1 cells and Th2 cells maintain balance (Th1/Th2 balance) by inhibiting each other's activation by cytokines produced by each, but for some reason, the induction of Th2 cells enhances symptoms of allergic diseases.

此情形時,推測因為產生活性氧之NADPH氧化酶的次單元(P40phox等)或分泌發炎性細胞激素之巨噬細胞等之發炎指標(F4/80等)的上昇,促進在生物體內的發炎性的細胞激素的分泌,而誘導過敏症狀。因此,藉由抑制 NADPH氧化酶或巨噬細胞等之發炎指標的上昇,而抑制發炎性的細胞激素的分泌,藉此能夠改善過敏症疾病。 In this case, it is presumed that the inflammatory index (F4/80, etc.) of the secondary unit of the active oxygen-containing NADPH oxidase (P40phox or the like) or the macrophage secreting the inflammatory cytokine (F4/80, etc.) is promoted to promote inflammation in the living body. The secretion of cytokines, while inducing allergy symptoms. Therefore, by suppressing An increase in the inflammatory index of NADPH oxidase or macrophages, and inhibition of the secretion of inflammatory cytokines, thereby improving allergic diseases.

作為針對過敏症疾病的醫藥,已知有抗組織胺、類固醇等。但,使用該等醫藥品時,有時伴隨耳鳴、頭痛、食欲不振等之副作用。又,該等物質從安全性及成本等方面考量,目前處於無法添加於飲食品的狀況。再者,有長期間投予少量過敏原而漸漸地弱化反應的減敏療法(hyposensitization),與為了避開與過敏原之接觸的膳食療法(dietotherapy)等,但關於該等,有需經過長期治療的必要性或降低生活品質(QOL)等問題,故期待開發出以經口攝取之簡便的方式而可期待預防過敏症疾病或治療效果的飲食品或飼料。 As a medicine for allergic diseases, antihistamines, steroids, and the like are known. However, when these medicines are used, there are some side effects such as tinnitus, headache, and loss of appetite. Moreover, these substances are considered in terms of safety and cost, and are currently in a state in which they cannot be added to foods and drinks. Furthermore, there is a hyposensitization which gradually weakens the reaction by administering a small amount of allergens for a long period of time, and a dietotherapy for avoiding contact with allergens, but there is a need for a long period of time. In view of the necessity of treatment or the reduction of quality of life (QOL), it is expected to develop a food or drink or a feed which can be expected to prevent allergic diseases or therapeutic effects by a simple method of oral intake.

作為改善過敏症狀的食品成分,已知有難消化性寡糖或類黃酮類(flavonoid)、多元不飽和脂肪酸等。已知難消化性寡糖,在將以比菲德氏菌作為主體之腸內菌叢的狀態保持於正常方面係為有效,並已知藉由攝取係難消化性寡糖之棉籽糖,會誘導Th1環境且同時能夠抑制全身免疫系的Th2反應。又,酸性木寡糖(Xylooligosaccharide)對於發炎性細胞激素的抑制效果亦已被認定,推測寡糖的攝取從各式各樣的方面展現抑制過敏症的效果。再者,關於以通式Gal-(Gal)n-Glc(n=1~3、β-1,4鍵結)表示之半乳寡醣(4’-半乳糖基乳糖(4’-Galactosyl Lactose)),已被報告經由抑制IgE的產生而有改善過敏症之作用。 As a food component which improves allergy symptoms, an indigestible oligosaccharide, a flavonoid, a polyunsaturated fatty acid, etc. are known. It is known that indigestible oligosaccharides are effective in maintaining the state of the intestinal flora mainly composed of F. faecalis, and it is known that by ingesting the cottonseed sugar which is an indigestible oligosaccharide, It induces the Th1 environment and at the same time inhibits the Th2 response of the systemic immune system. Further, the inhibitory effect of Xylooligosaccharide on inflammatory cytokines has also been confirmed, and it is presumed that the intake of oligosaccharides exhibits an effect of suppressing allergy from various aspects. Furthermore, the galactooligosaccharide (4'-Galactosyl Lactose) represented by the general formula Gal-(Gal)n-Glc (n=1~3, β-1,4 linkage) )) has been reported to have an effect of improving allergy by inhibiting the production of IgE.

【先前技術文獻】 [Previous Technical Literature]

【專利文獻】 [Patent Literature]

專利文獻1:日本特開2007-197331號公報 Patent Document 1: Japanese Laid-Open Patent Publication No. 2007-197331

專利文獻2:日本特開2001-288093號公報 Patent Document 2: Japanese Laid-Open Patent Publication No. 2001-288093

專利文獻3:日本特開2007-23018號公報 Patent Document 3: JP-A-2007-23018

專利文獻4:日本特開昭62-79791號公報 Patent Document 4: Japanese Laid-Open Patent Publication No. 62-79791

【非專利文獻】 [Non-patent literature]

非專利文獻1:British Joumal of Nutrition,88,421-426,2002 Non-Patent Document 1: British Joumal of Nutrition, 88, 421-426, 2002

非專利文獻2:Journal of Nutritional Science and Vitaminology, 56, 54-59, 2010 Non-Patent Document 2: Journal of Nutritional Science and Vitaminology, 56, 54-59, 2010

本發明之課題係提供一種過敏症改善劑,藉由抑制發炎性的細胞激素的分泌改善生物體內的免疫調節系統的混亂。 An object of the present invention is to provide an allergy-improving agent which improves the disorder of an immune regulatory system in a living body by inhibiting the secretion of an inflammatory cytokine.

本案發明人等,為解決前述課題致力進行探討時,發現具有以O-β-D-半乳呋喃糖基-(1→6)-[O-β-D-半乳呋喃糖基-(1→4)]-D-葡萄糖或O-β-D-半乳呋喃糖基-(1→4)-[O-β-D-半乳呋喃糖基-(1→6)]-D-葡萄糖表示之結構式之半乳寡醣,經由抑制發炎性細胞激素的分泌而有過敏症改善作用。 The inventors of the present invention have found that O-β-D-galactofuranosyl-(1→6)-[O-β-D-galactofuranosyl-(1) has been found to solve the above problems. →4)]-D-glucose or O-β-D-galactofuranosyl-(1→4)-[O-β-D-galipofuranosyl-(1→6)]-D-glucose The structural galactooligosaccharide represented by the formula has an allergic effect by inhibiting the secretion of inflammatory cytokines.

亦即本發明,係包含以下之態樣者。 That is, the present invention includes the following aspects.

(1)一種過敏症改善劑,係以具有O-β-D-半乳呋喃糖基-(1→6)-[O-β-D-半乳呋喃糖基-(1→4)]-D-葡萄糖或O-β-D-半乳呋喃糖基-(1→4)-[O-β-D-半乳呋喃糖基-(1→6)]-D-葡萄糖表示之結構式之半乳寡醣作為有效成分。 (1) An allergy-improving agent having O-β-D-galactofuranosyl-(1→6)-[O-β-D-galactofuranosyl-(1→4)]- D-glucose or O-β-D-galactofuranosyl-(1→4)-[O-β-D-galipofuranosyl-(1→6)]-D-glucose A galactooligosaccharide is used as an active ingredient.

(2)如(1)記載之過敏症改善劑,其中,半乳寡醣係來自哺乳類的乳汁。 (2) The allergy-improving agent according to (1), wherein the galactooligosaccharide is derived from a mammalian milk.

(3)如(1)或(2)記載之過敏症改善劑,其中,半乳寡醣係來自牛乳。 (3) The allergy-improving agent according to (1) or (2), wherein the galactooligosaccharide is derived from cow's milk.

(4)一種過敏症改善用飲食品、過敏症改善用營養組成物、過敏症改善用飼料或過敏症改善用醫藥品,含有如(1)至(3)項中之任一項記載之過敏症改善劑。 (4) A food or drink for improving allergy, a nutritional composition for allergy improvement, a food for allergy improvement, or a drug for improving allergy, containing the allergy according to any one of (1) to (3) Disease improver.

(5)一種過敏症的改善方法,係藉由經口攝取如(1)至(3)項中之任一項記載之過敏症改善劑。 (5) A method for improving allergy, which is an allergy-improving agent according to any one of (1) to (3).

本發明之過敏症改善劑,藉由抑制代謝性的發炎並抑制發炎性的細胞 激素的分泌以此改善生物體內的免疫調節系統的混亂。 The allergy improving agent of the present invention inhibits metabolic inflammation and inhibits inflammatory cells The secretion of hormones improves the confusion of the immune regulation system in the body.

以下,舉出實施態樣進行本發明之說明,但本發明並不限定於以下實施態樣。 Hereinafter, the present invention will be described by way of embodiments, but the present invention is not limited to the following embodiments.

[過敏症改善劑] [allergy improvement agent]

本發明之過敏症改善劑係以具有O-β-D-半乳呋喃糖基-(1→6)-[O-β-D-半乳呋喃糖基-(1→4)]-D-葡萄糖或O-β-D-半乳呋喃糖基-(1→4)-[O-β-D-半乳呋喃糖基-(1→6)]-D-葡萄糖表示之結構式之半乳寡醣作為有效成分。 The allergy improving agent of the present invention has O-β-D-galiptooligosaccharyl-(1→6)-[O-β-D-galiptofuranosyl-(1→4)]-D- Glucose or O-β-D-galactofuranosyl-(1→4)-[O-β-D-galipofuranosyl-(1→6)]-D-glucose Oligosaccharides are used as active ingredients.

以前述半乳寡醣作為有效成分之過敏症改善劑,可抑制NADPH氧化酶或巨噬細胞等發炎指標的上昇,抑制發炎性的細胞激素的分泌。藉此,在預防及/或治療異位性皮膚炎、過敏症性鼻炎、蕁麻疹、花粉症等之各種過敏症疾病係為有用。又,於預防及/或治療風溼病(Rheumatism)亦為有用。 The allergy-improving agent containing the galactooligosaccharide as an active ingredient can suppress an increase in inflammatory indexes such as NADPH oxidase or macrophages, and inhibit the secretion of inflammatory cytokines. Therefore, it is useful for preventing and/or treating various allergic diseases such as atopic dermatitis, allergic rhinitis, urticaria, and hay fever. Further, it is also useful for preventing and/or treating rheumatism.

再者,記載於本申請案之改善劑所使用的半乳寡醣係以乳糖作為原料,故能簡便且經濟性地進行製造,又,有效成分之糖構成已被特定,故就安全性或效力方面而言能夠充分地進行測試或探討,因此有優秀的穩定性及安全性,再者,展現優良的過敏症改善效果的同時,有優秀的穩定性及安全性,故也有可迅速及容易地應用在食品或醫藥之效果。 Further, the galactooligosaccharide used in the improvement agent of the present application uses lactose as a raw material, so that it can be produced simply and economically, and the sugar composition of the active ingredient is specified, so that it is safe or In terms of efficacy, it can be fully tested or explored, so it has excellent stability and safety. In addition, it exhibits excellent allergy improvement effects, excellent stability and safety, and therefore can be quickly and easily The effect of applying it on food or medicine.

作為有效成分使用之具有以O-β-D-半乳呋喃糖基-(1→6)-[O-β-D-半乳呋喃糖基-(1→4)]-D-葡萄糖或O-β-D-半乳呋喃糖基-(1→4)-[O-β-D-半乳呋喃糖基-(1→6)]-D-葡萄糖表示之結構式之半乳寡醣,例如可藉由以下方式獲得:使Sumirakuto LL等之市售β-半乳糖苷酶(galactosidase)在含有乳糖20%~40%之溶液中反應數小時後,進行減壓濃縮去除未反應之乳糖,將獲得的溶液通過活性碳管柱,並以20%乙醇溶出吸附之半乳寡醣。又,可藉由如記載於專利文獻4之以下方式獲得:將屬於麴菌屬或青黴菌屬、木黴菌屬之乳糖利用性菌在乳糖含量約3%、pH6的培養基進行預培養,藉此使其增殖,並將 經增殖之該乳糖利用性菌在乳糖含量約13%、pH4至9的主培養培養基進行主培養後,進行減壓濃縮從該培養液去除未反應之乳糖,再進一步使β-半乳糖苷酶進行作用,分解培養液中殘留之未反應的乳糖,再將獲得的溶液通過活性碳管柱,並以20%乙醇溶出吸附之半乳寡醣。再者,此情形時,若將主培養以20至40℃,更佳為以30℃左右進行5至15日的振盪培養,則可更為提高產率。 As an active ingredient, it has O-β-D-galofuranosyl-(1→6)-[O-β-D-galipuranosyl-(1→4)]-D-glucose or O -β-D-galactofuranosyl-(1→4)-[O-β-D-galipofuranosyl-(1→6)]-D-glucose is a structural galactooligosaccharide, For example, it can be obtained by reacting commercially available β-galactosidase of Sumirakuto LL or the like in a solution containing 20% to 40% of lactose for several hours, and then concentrating under reduced pressure to remove unreacted lactose. The obtained solution was passed through an activated carbon tube column, and the adsorbed galactooligosaccharide was eluted with 20% ethanol. Further, it can be obtained by the following method as described in Patent Document 4: a lactose-utilizing bacterium belonging to the genus Trichophyton, Penicillium or Trichoderma is pre-cultured in a medium having a lactose content of about 3% and pH 6 Make it proliferate and The lactose-utilizing bacteria which have been proliferated are subjected to main culture in a main culture medium having a lactose content of about 13% and a pH of 4 to 9, and then concentrated under reduced pressure to remove unreacted lactose from the culture solution, and further to make β-galactosidase The action is performed to decompose the unreacted lactose remaining in the culture solution, and the obtained solution is passed through an activated carbon tube column, and the adsorbed galactooligosaccharide is eluted with 20% ethanol. Further, in this case, if the main culture is shaken at 20 to 40 ° C, more preferably at about 30 ° C for 5 to 15 days, the yield can be further improved.

又,也能夠將以如前述方式製備之半乳寡醣以逆滲透膜(RO)等進行濃縮,或藉由冷凍乾燥等進行乾燥。 Further, the galactooligosaccharide prepared as described above may be concentrated by a reverse osmosis membrane (RO) or the like, or may be dried by freeze drying or the like.

作為半乳寡醣原料之乳糖係指由牛或山羊、綿羊、人類等之哺乳類的乳汁製備之乳糖,或其凝集物、粉末、或者精製之乳糖;使用此與β-半乳糖苷酶進行反應的情形時,或進行主培養的情形時能夠以水溶液的狀態進行使用。 Lactose, which is a raw material for galactooligosaccharides, refers to lactose prepared from the milk of mammals such as cattle or goats, sheep, humans, etc., or agglomerates, powders, or refined lactose; using this to react with β-galactosidase In the case of the case of the main culture, it can be used in the state of an aqueous solution.

[含有過敏症改善劑之飲食品、飼料、醫藥品等之過敏症改善用產品] [Products for allergy improvement of foods, foods, medicines, etc. containing allergy-improving agents]

作為本發明的實施態樣,也可將前述半乳寡醣直接作為過敏症改善劑進行使用,也可視須要根據常法,製劑化為粉末劑、顆粒劑、錠劑、膠囊劑、飲用劑等進行利用。又,關於進一步藉由逆滲透膜(RO)處理等獲得之半乳寡醣,也可直接作為過敏症改善劑進行使用,也可直接乾燥進行使用。又,也可按照常法,進行製劑化使用。 As an embodiment of the present invention, the galactooligosaccharide may be directly used as an allergy-improving agent, or may be formulated into a powder, a granule, a tablet, a capsule, a drink, etc. according to a usual method. Use it. Further, the galactooligosaccharide obtained by further reverse osmosis membrane (RO) treatment or the like may be used as an allergy-improving agent as it is, or may be directly dried and used. Moreover, it can also be formulated and used according to a normal method.

再者,在將該等製劑化後,也可將其摻合在營養劑或酸酪乳、飲料、薄餅乾等飲食品、營養組成物、飼料及醫藥品。 Further, after the preparation is carried out, it may be blended into a food or drink, a nutritional composition, a feed, and a pharmaceutical such as a nutrient or yogurt, a drink, or a thin biscuit.

本說明書中之「過敏症改善用飲食品、過敏症改善用營養組成物、過敏症改善用飼料及過敏症改善用醫藥品」,除了僅包含有效成分之半乳寡醣的情形以外,還可含有安定劑或糖類、脂質、香味料、維他命、礦物質、類黃酮、多酚等其他飲食品、飼料及醫藥品中通常含有的原材料等。又,除了有效成分半乳寡醣以外,也可與其他的展現過敏症改善作用的成分,例如棉籽糖或酸性木寡糖等一同使用。 In the present specification, the "food for allergy improvement, the nutritional composition for allergy improvement, the food for allergy improvement, and the medicine for allergy improvement" may be in addition to the galactooligosaccharide containing only the active ingredient. Contains stabilizers or sugars, flavors, vitamins, minerals, flavonoids, polyphenols and other foods and beverages, raw materials commonly found in feeds and pharmaceuticals. Further, in addition to the active ingredient galactooligosaccharide, it may be used together with other components exhibiting an allergy-improving action, such as raffinose or acid xylooligosaccharide.

又,也可將此類過敏症改善用飲食品、過敏症改善用營養組成物、過 敏症改善用飼料或過敏症改善用醫藥品作為原材料,摻合通常包含於其他飲食品等之原材料等進行製備。 In addition, it is also possible to use such foods and foods for allergy improvement and nutrient composition for allergy improvement. The sensitization improving feed or the allergy-improving pharmaceutical product is used as a raw material, and is prepared by blending a raw material or the like which is usually included in other foods and drinks.

過敏症改善用飲食品、過敏症改善用營養組成物、過敏症改善用飼料及過敏症改善用醫藥品中之半乳寡醣的摻合量,無特別限制,為了讓成人每人每日經口攝取半乳寡醣在50mg以上,雖然依據飲食品、飼料及醫藥品的態樣有所不同,但一般較佳為相對於全部質量含有0.01~10%(質量/質量),更佳為含有0.1~5%(質量/質量)。 There is no particular limitation on the amount of galacto-oligosaccharide in the food for allergy improvement, the nutritional composition for allergy improvement, the feed for allergy improvement, and the medicinal product for allergy improvement. The oral intake of galacto-oligosaccharides is 50 mg or more, and although it is different depending on the aspect of foods, drinks, and pharmaceuticals, it is generally preferably 0.01 to 10% (mass/mass), more preferably contained, based on the total mass. 0.1~5% (quality/quality).

記載於本發明之過敏症改善劑可在前述的有效成分中添加適當的助劑而製劑化成任意的態樣,作為能夠進行口服投予之過敏症改善用組成物。在製劑化時,可使用通常使用之填充劑、增量劑、黏結劑、崩解劑、界面活性劑、潤滑劑等稀釋劑或賦形劑。作為賦形劑可將例如蔗糖、乳糖、澱粉、結晶性纖維素、甘露醇、輕質矽酸酐、鋁酸鎂、合成矽酸鋁、偏矽酸鋁酸鎂、碳酸鈣、碳酸氫鈉、磷酸氫鈣、羧基甲基纖維素鈣等1種或2種以上予以組合而加入。 The allergy-improving agent of the present invention can be formulated into an arbitrary form by adding an appropriate auxiliary agent to the above-mentioned active ingredient, and can be used as an allergy-improving composition capable of oral administration. At the time of formulation, a diluent or an excipient such as a filler, a bulking agent, a binder, a disintegrating agent, a surfactant, a lubricant, or the like which are generally used can be used. As excipients, for example, sucrose, lactose, starch, crystalline cellulose, mannitol, light phthalic anhydride, magnesium aluminate, synthetic aluminum citrate, magnesium metasilicate aluminate, calcium carbonate, sodium hydrogencarbonate, phosphoric acid One type or two or more types of hydrogen calcium and carboxymethyl cellulose calcium are added in combination.

[過敏症的改善方法] [Improvement of allergy]

本發明也係關於將上述實施態樣之過敏症改善劑經口攝取之過敏症改善方法。該過敏症改善方法,對於各種過敏症疾病的治療及/或預防係為有用。該過敏症改善方法對於哺乳類例如人類,及其他犬、貓等的家畜有效果。 The present invention also relates to an allergy-improving method for orally ingesting an allergy-improving agent according to the above embodiment. This allergy improvement method is useful for the treatment and/or prevention of various allergic diseases. This allergy improvement method is effective for mammals such as humans and other domestic animals such as dogs and cats.

【實施例】 [Examples]

以下表示實施例、測試例,針對本發明進行詳細說明,但該等僅為例示而已,本發明並不因該等受到任何限定。 The present invention is described in detail below with reference to the embodiments and the examples of the invention, but these are merely illustrative, and the invention is not limited thereto.

[實施例1] [Example 1]

添加市售之β-半乳糖苷酶(商品名:Sumirakuto LL、新日本化學工業(股))至20%乳糖水溶液5000ml使其成為3單位/ml,在40℃(pH5.0)進行反應3小時。 將此溶液進行減壓濃縮析出乳糖並予以去除,通入至活性碳管柱(15cm×30cm)吸附半乳寡醣後,通入5%乙醇3000ml,以去除殘留之乳糖。其次,通入20%乙醇3000ml,將所獲得的溶出級分進行減壓濃縮後,進行冷凍乾燥而獲得有效成分之半乳寡醣之粉末(實施例品1)205g。於此半乳寡醣粉末包含12質量%之具有以O-β-D-半乳呋喃糖基-(1→6)-[O-β-D-半乳呋喃糖基-(1→4)]-D-葡萄糖或O-β-D-半乳呋喃糖基-(1→4)-[O-β-D-半乳呋喃糖基-(1→6)]-D-葡萄糖表示之結構式之半乳寡醣。 A commercially available β-galactosidase (trade name: Sumirakuto LL, Nippon Chemical Industry Co., Ltd.) was added to 5000 ml of a 20% aqueous lactose solution to make 3 units/ml, and the reaction was carried out at 40 ° C (pH 5.0). hour. The solution was concentrated under reduced pressure to precipitate and remove lactose, and the galactose oligosaccharide was adsorbed to a column of activated carbon tubes (15 cm × 30 cm), and then 3,000 ml of 5% ethanol was introduced to remove residual lactose. Next, 3000 ml of 20% ethanol was introduced, and the obtained eluted fraction was concentrated under reduced pressure, and then lyophilized to obtain 205 g of a powder of galactooligosaccharide (Example 1) of an active ingredient. The galactooligosaccharide powder contains 12% by mass of O-β-D-galofuranosyl-(1→6)-[O-β-D-galactofuranosyl-(1→4) ]-D-glucose or O-β-D-galactofuranosyl-(1→4)-[O-β-D-galipofuranosyl-(1→6)]-D-glucose structure A galactooligosaccharide.

以如此方式獲得之半乳寡醣,能夠直接作為過敏症改善劑進行使用。 The galactooligosaccharide obtained in this manner can be directly used as an allergy improving agent.

[實施例2] [Embodiment 2]

將藍酪黴菌(Penicillium roqueforti)IFO 4135株接種至預培養培養基(乳糖2.9%、磷酸氫二鉀0.1%、硫酸鎂七水合物0.05%、硝酸鈉0.2%、氯化鉀0.05%、硫酸鐵0.001%)100ml,在30℃培養5天。將所獲得的預培養液100ml,接種至主培養培養基(乳糖13%、磷酸氫二鉀0.09%、硫酸鎂七水合物0.04%、硝酸鈉0.17%、氯化鉀0.04%、硫酸鐵0.001%)5000ml,在30℃振盪培養10天。將此培養液進行離心分離去除菌體後,進行減壓濃縮析出並去除培養液中之乳糖。將其溶液通入至活性碳管柱(15cm×30cm)吸附半乳寡醣後,通入5%乙醇3000ml,去除殘留之乳糖。其次,通入20%乙醇3000ml,將所獲得的溶出級分進行減壓濃縮後,進行冷凍乾燥而獲得半乳寡醣之粉末(實施例品2)50g。於此半乳寡醣粉末包含9質量%之具有以O-β-D-半乳呋喃糖基-(1→6)-[O-β-D-半乳呋喃糖基-(1→4)]-D-葡萄糖或O-β-D-半乳呋喃糖基-(1→4)-[O-β-D-半乳呋喃糖基-(1→6)]-D-葡萄糖表示之結構式之半乳寡醣。 Penicillium roqueforti IFO 4135 strain was inoculated to pre-culture medium (lactose 2.9%, dipotassium hydrogen phosphate 0.1%, magnesium sulfate heptahydrate 0.05%, sodium nitrate 0.2%, potassium chloride 0.05%, iron sulfate 0.001) %) 100 ml, cultured at 30 ° C for 5 days. 100 ml of the obtained preculture solution was inoculated to the main culture medium (lactose 13%, dipotassium hydrogen phosphate 0.09%, magnesium sulfate heptahydrate 0.04%, sodium nitrate 0.17%, potassium chloride 0.04%, iron sulfate 0.001%) 5000 ml, cultured at 30 ° C for 10 days with shaking. After the culture solution was centrifuged to remove the cells, the solution was concentrated under reduced pressure to remove the lactose in the culture solution. After the solution was passed through a column of activated carbon tubes (15 cm × 30 cm) to adsorb galactooligosaccharides, 3000 ml of 5% ethanol was introduced to remove residual lactose. Next, 3000 ml of 20% ethanol was introduced, and the obtained eluted fraction was concentrated under reduced pressure, and then lyophilized to obtain 50 g of a galactooligosaccharide powder (Example 2). The galactooligosaccharide powder contains 9% by mass of O-β-D-galactofuranosyl-(1→6)-[O-β-D-galactofuranosyl-(1→4) ]-D-glucose or O-β-D-galactofuranosyl-(1→4)-[O-β-D-galipofuranosyl-(1→6)]-D-glucose structure A galactooligosaccharide.

以如此方式獲得的半乳寡醣,能夠直接作為過敏症改善劑進行使用。 The galactooligosaccharide obtained in this manner can be directly used as an allergy improving agent.

[測試例1] [Test Example 1]

(動物實驗) (Animal experiment)

使用實施例品1、2之半乳寡醣,針對過敏症改善作用進行評價。實驗使用4週大的BALB/c小鼠。在小鼠的右耳的內側塗佈10μl的0.3%的2,4,6-三硝基-1-氯苯(以下稱為「TNCB」)溶液,使其進行致敏作用(sensitization)。1週後,每隔2至3日塗佈10μl的0.3%TNCB溶液在小鼠的右耳的內側。與上 述TNCB溶液的重複塗佈並行,每日進行下述口服投予連續2週:期間對於A群組(對照)、B群組、C群組(各群組各8隻)的小鼠,於A群組使用生理食鹽水,於B群組小鼠體重每1kg使用50mg實施例品1的半乳寡醣,於C群組小鼠體重每1kg使用50mg實施例品2的半乳寡醣。在B群組、C群組,使實施例品1、2懸浮於生理食鹽水而進行口服投予。測試結束時,摘除小鼠右耳測定質量,同時使用即時PCR方式測定發炎指標(F4/80、P40phox)的mRNA表現量。其結果表示於表1。 The allergy-improving effects were evaluated using the galactooligosaccharides of Examples 1 and 2. The experiment used 4 week old BALB/c mice. 10 μl of a 0.3% solution of 2,4,6-trinitro-1-chlorobenzene (hereinafter referred to as "TNCB") was applied to the inner side of the right ear of the mouse to cause sensitization. One week later, 10 μl of a 0.3% TNCB solution was applied to the inside of the right ear of the mouse every 2 to 3 days. With The repeated application of the TNCB solution was performed in parallel, and the following oral administration was carried out for 2 consecutive weeks: during the period of the A group (control), the B group, and the C group (8 in each group), In group A, physiological saline was used, and 50 mg of the galactooligosaccharide of Example 1 was used per 1 kg of body weight of the B group mice, and 50 mg of the galactooligosaccharide of Example 2 was used for 1 kg of the body weight of the C group mice. In the group B and the group C, the sample products 1 and 2 were suspended in physiological saline and administered orally. At the end of the test, the quality of the right ear was measured and the mRNA expression of the inflammatory index (F4/80, P40phox) was determined by real-time PCR. The results are shown in Table 1.

實驗結果,投予2週後的右耳的質量或發炎指標(F4/80、P40phox)的mRNA表現量,與A群組(對照)相較,在小鼠體重每1kg分別口服投予半乳寡醣50mg以上之實施例品1、2之B、C群組顯著降低。由此結果,得知實施例品1、2的半乳寡醣有過敏症改善作用。又,可知此改善作用在對小鼠體重每1kg投予實施例品1、2的半乳寡醣50mg以上之情形時會觀測到。 The results of the experiment showed that the quality of the right ear or the index of inflammatory index (F4/80, P40phox) after 2 weeks of administration was compared with that of the A group (control), and the half-milk was orally administered per 1 kg of the mouse body weight. The group of B and C of the exemplified product 1 and 2 having an oligosaccharide of 50 mg or more was remarkably lowered. From the results, it was found that the galactooligosaccharides of Examples 1 and 2 had an allergy-improving effect. Further, it was found that this improvement was observed when 50 parts of galactooligosaccharides of Examples 1 and 2 were administered to 1 kg of mouse body weight.

[實施例3] [Example 3]

(過敏症改善用錠劑之製備) (Preparation of lozenges for allergy improvement)

以表2表示之配比,混合各原料使其成為合計為100.0質量份後,依常法成形、打錠為1g,而製造過敏症改善用錠劑。 In the compounding ratio shown in Table 2, the raw materials were mixed to a total of 100.0 parts by mass, and then formed into a total of 1 g, and an ingot for improving allergy was produced.

[實施例4] [Example 4]

(過敏症改善用液狀營養組成物之製備) (Preparation of liquid nutritional composition for allergy improvement)

將實施例品2的半乳寡醣50g溶解於4,950g的去離子水,加熱至50℃後,以TK均質混合機(TK ROBO MICS;特殊機化工業公司製),以6,000rpm攪拌混合30分鐘,而獲得含有實施例品2之半乳寡醣含量50g/5kg之半乳寡醣溶液。於此半乳寡醣溶液5.0kg中,摻合酪蛋白5.0kg、大豆蛋白質5.0kg、魚油1.0kg、紫蘇油3.0kg、糊精17.0kg、礦物質混合物6.0kg、維他命混合物1.95kg、乳化劑2.0kg、安定劑4.0kg、香料0.05kg,並填充至200ml的殺菌軟袋內,以蒸餾殺菌機(第1種壓力容器、TYPE:RCS-4CRTGN、日阪製作所製)於121℃殺菌20分鐘,製成過敏症改善用液狀營養組成物50kg。 50 g of galactooligosaccharide of Example 2 was dissolved in 4,950 g of deionized water, and the mixture was heated to 50 ° C, and then stirred and mixed at 6,000 rpm with a TK homomixer (TK ROBO MICS; manufactured by Special Machine Chemical Co., Ltd.). In a minute, a galactooligosaccharide solution containing 50 g/5 kg of the galactooligosaccharide content of Example 2 was obtained. In 5.0 kg of this galactooligosaccharide solution, 5.0 kg of casein, 5.0 kg of soybean protein, 1.0 kg of fish oil, 3.0 kg of perilla oil, 17.0 kg of dextrin, 6.0 kg of mineral mixture, 1.95 kg of vitamin mixture, emulsifier 2.0 kg, a stabilizer of 4.0 kg, and a fragrance of 0.05 kg, and filled in a 200 ml sterilized soft bag, and sterilized at 121 ° C for 20 minutes in a distillation sterilizer (first type pressure vessel, TYPE: RCS-4 CRTGN, manufactured by HISAKA CO., LTD.). 50 kg of a liquid nutritional composition for allergy improvement was prepared.

[實施例5] [Example 5]

(過敏症改善用飲料之製備) (Preparation of beverages for allergy improvement)

將脫脂奶粉300g溶解至409g之去離子水後,溶解實施例品1之半乳寡醣1g,並加熱至50℃後,以超分散器(ULTRA-TURRAXT-25;IKA日本公司製),用9,500rpm攪拌混合30分鐘。添加麥芽糖醇100g、酸味料2g、還原糖漿20g、香料2g、去離子水166g後,填充至100ml的玻璃瓶中,於95℃殺菌15秒鐘後蓋緊,而製成過敏症改善用飲料10瓶(內容量100ml)。 After dissolving 300 g of skim milk powder in 409 g of deionized water, 1 g of the galactooligosaccharide of Example 1 was dissolved, and the mixture was heated to 50 ° C, and then used as a super disperser (ULTRA-TURRAXT-25; manufactured by IKA Japan Co., Ltd.). Mix and mix for 30 minutes at 9,500 rpm. After adding 100 g of maltitol, 2 g of sourdil, 20 g of reducing syrup, 2 g of flavoring agent, and 166 g of deionized water, the mixture was filled in a 100 ml glass bottle, sterilized at 95 ° C for 15 seconds, and then tightly closed to prepare an allergy-improving beverage 10 . Bottle (content amount 100ml).

[實施例6] [Embodiment 6]

(犬用過敏症改善飼料的製備) (Canine allergy improves feed preparation)

將實施例品2之半乳寡醣2kg溶解於98kg的去離子水,並加熱至50 ℃後,以TK均質混合機(MARK II 160型;特殊機化工業公司製),用3,600rpm攪拌混合40分鐘,而獲得含有實施例品2之半乳寡醣2g/100g的半乳寡醣溶液。於此半乳寡醣溶液10kg摻合大豆粕12kg、脫脂奶粉14kg、黃豆油4kg、玉米油2kg、棕櫚油23.2kg、玉米澱粉14kg、小麥粉9kg、麥麩2kg、維他命混合物5kg、纖維素2.8kg、礦物質混合物2kg,於120℃殺菌4分鐘,而製成犬用過敏症改善飼料100kg。 2 kg of galactooligosaccharide of Example 2 was dissolved in 98 kg of deionized water and heated to 50 After the temperature was °C, the mixture was stirred and mixed at 3,600 rpm for 40 minutes with a TK homomixer (manufactured by MARK II 160; manufactured by Kosei Kogyo Co., Ltd.) to obtain a galactooligosaccharide containing 2 g/100 g of the galactooligosaccharide of Example 2. Solution. 10 kg of this galactooligosaccharide solution was blended with soybean meal 12 kg, skim milk powder 14 kg, soybean oil 4 kg, corn oil 2 kg, palm oil 23.2 kg, corn starch 14 kg, wheat flour 9 kg, wheat bran 2 kg, vitamin mixture 5 kg, cellulose 2.8 2 kg of kg and mineral mixture were sterilized at 120 ° C for 4 minutes to prepare 100 kg of dog allergy-improving feed.

Claims (5)

一種過敏症改善劑,以具有O-β-D-半乳呋喃糖基-(1→6)-[O-β-D-半乳呋喃糖基-(1→4)]-D-葡萄糖或O-β-D-半乳呋喃糖基-(1→4)-[O-β-D-半乳呋喃糖基-(1→6)]-D-葡萄糖表示之結構式之半乳寡醣作為有效成分。 An allergy improving agent having O-β-D-galipofuranosyl-(1→6)-[O-β-D-galipuranosyl-(1→4)]-D-glucose or O-β-D-galofuranosyl-(1→4)-[O-β-D-galipofuranosyl-(1→6)]-D-glucose is a structural galactooligosaccharide As an active ingredient. 如申請專利範圍第1項之過敏症改善劑,其中,該半乳寡醣係來自哺乳類之乳汁。 The allergy-improving agent according to claim 1, wherein the galactooligosaccharide is derived from a mammalian milk. 如申請專利範圍第1或2項之過敏症改善劑,其中,該半乳寡醣係來自牛乳。 The allergy-improving agent according to claim 1 or 2, wherein the galactooligosaccharide is derived from cow's milk. 一種過敏症改善用飲食品、過敏症改善用營養組成物、過敏症改善用飼料或過敏症改善用醫藥品,其特徵在於含有如申請專利範圍第1至3項中之任一項之過敏症改善劑。 A food or drink for improving allergy, a nutritional composition for improving allergy, a feed for allergy improvement, or a pharmaceutical for allergy improvement, characterized by containing an allergy as in any one of claims 1 to 3. Improver. 一種過敏症改善方法,其特徵在於經口攝取如申請專利範圍1至3項中之任一項之過敏症改善劑。 An allergy-improving method characterized by orally ingesting an allergy-improving agent according to any one of claims 1 to 3.
TW102142097A 2012-11-19 2013-11-19 Allergy improving agent TW201427603A (en)

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DE102020133860A1 (en) 2020-12-16 2022-06-23 MNT Systeme GmbH USE OF β-GALACTOOLIGOSACCHRIDS FOR THE PREPARATION OF A COSMETIC AND/OR PHARMACOLOGICALLY ACTIVE COMPOSITION

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