CN102006788A - 合益素混合物 - Google Patents
合益素混合物 Download PDFInfo
- Publication number
- CN102006788A CN102006788A CN2009801129835A CN200980112983A CN102006788A CN 102006788 A CN102006788 A CN 102006788A CN 2009801129835 A CN2009801129835 A CN 2009801129835A CN 200980112983 A CN200980112983 A CN 200980112983A CN 102006788 A CN102006788 A CN 102006788A
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- CN
- China
- Prior art keywords
- preparation
- oligosaccharides
- lactobacillus
- acetyl group
- lactose
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
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- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
本发明涉及包含N-乙酰基-乳糖胺和/或含有N-乙酰基-乳糖胺的寡糖和益生菌乳杆菌属的物种的制剂。本发明扩展至包含所述制剂的营养组合物,以及所述制剂在预防和治疗胃肠道和上呼吸道病原性感染中的用途。
Description
发明领域
本发明涉及包含益生菌和益生元寡糖的制剂,其特异性地设计为增强益生菌的功效,并且涉及包含所述制剂的食物产品和所述制剂的用途。
发明背景
对所有的婴儿都建议使用母乳。但是,在某些情况下,由于医学的原因母乳喂养不足或不成功,或者母亲选择不进行母乳喂养。针对这些情况已开发了婴儿配方食品。
最近,已发现人们如果摄取某些菌株的话,这些菌株会呈现有价值的性质,这已引起了相当的重视。具体而言,已发现乳杆菌属(Lactobacilli)和双歧杆菌属(Bifidobacteria)的特定菌株能在肠道内建群,降低病原菌贴附肠上皮的能力,具有免疫调节效应并能够协助维持健康状态。有时将这样的细菌称为益生菌,并且已经提出将合适的益生菌加入到婴儿配方食品中。
已进行了广泛的研究来鉴定新型益生菌株。例如,EP 0 199 535、EP 0768 375、WO 97/00078、EP 0 577 903和WO 00/53200公开了乳杆菌属和双歧杆菌属的特定菌株及其有益的效果。
如前所述,由于能够在肠粘膜上建群,降低病原菌贴附肠上皮的能力,某些益生菌株已用于预防和治疗婴儿的腹泻。例如,已报道包括芬兰的Valio Oy在内生产的商品名为LGG的鼠李糖乳杆菌(Lactobacillus rhamnosus)ATCC 53103能有效减少细菌诱导的婴儿和儿童腹泻(Canani等,British Medical Journal 2007,Aug 18;335(7615):340),通常将其添加到人类食物、例如发酵乳制品中。
施用益生元,例如在食品中添加益生元是预防或治疗胃肠道细菌病原体例如大肠杆菌(EPEC)感染的另一种方法。益生元是不消化的食品成分,其通过选择性刺激结肠中一种或有限数量的细菌生长和/或活性来有益地影响宿主,从而改善宿主健康。此类成分是不消化的,因为其在胃或小肠中不能分解和吸收,从而可以完整通过到达结肠,在此处由有益细菌选择性发酵。益生元的实例包括某些寡糖,例如寡聚果糖(FOS)和半乳寡聚糖(GOS)。Shoaf等人研究了包括寡聚果糖、菊糖、半乳寡聚糖、乳果糖和棉子糖在内的多种益生元抑制EPEC菌株E2348/69粘附Hep-2和Caco-2细胞的能力,他们发现纯化的半乳寡聚糖对Hep-2和Caco-2细胞均具有最大粘附抑制,分别使EPEC的粘附下降了65和70%,并推断根据其观察结果表明某些益生元寡糖可能具有抗粘附活性,直接抑制了病原体对宿主上皮细胞表面的粘附(Infect Immun 2006 Dec;74(12):6920-8)。
已知人乳比大多数其他动物乳含有更多的不能消化的寡糖。事实上,不能消化的寡糖代表了母乳中的第三大固体成分(在乳糖和脂类之后),初乳中以12-15g/l的浓度存在,成熟乳中以5-8g/l存在。人乳的寡糖非常耐受酶水解,表明这些寡糖可能显示出与其热值不直接相关的关键性功能。
由于对人乳的组成已经有更好的了解,已经建议将益生元添加到婴儿配方食品中。多种补充了益生元例如寡聚果糖和半乳寡聚糖混合物的婴儿配方食品是可商购的。但是,此类混合物只是粗略的近似人乳中的寡糖混合物。在人乳中已经检测到超过100种不同的寡糖成分,其中一些尚未在动物乳例如牛乳中检测到,或仅检测到少量。仅以非常小的量存在于牛乳和初乳中或完全不存在的人乳寡糖类型的实例是唾液酸化和岩藻糖基化的寡糖。
尽管某些寡糖已证实与降低病原体结合宿主上皮细胞能力有关,但各种不同寡糖的数目和功能依然有待阐明。例如,Cravioto等人报道人乳中寡糖富集的组分抑制了EPEC对HEp-2细胞的粘附(The Journal of Infectious Diseases 1991;163:1247-1255)。
对生产婴儿配方食品的持续呼声还建议婴儿配方食品既含益生菌又含益生元,其尽可能的复制接近人乳的组成和功效。例如,在WO 2007/101675中,建议在婴儿配方食品中补充益生菌菌株混合物和N-乙酰化、中性和唾液酸化的寡糖混合物,所述混合物提供比现有市售的寡聚果糖和半乳寡聚糖混合物更接近于人乳中的寡糖组成。据称,被描述为合益素(synbiotic)的该混合物可用于预防病原性感染。
然而,仍然一直都需要通过组合具有特殊有益效果的特定益生菌和益生元来进一步改善婴儿配方食品等组合物的保护效应。
发明概述
现在令人惊讶的发现,N-乙酰基-乳糖胺和/或含有N-乙酰基-乳糖胺的寡糖的施用在提高与所述寡糖共同施用的益生菌乳杆菌属的物种(Lactobacillus sp)的有益效果和功效方面是非常有效的。
因此,本发明在第一方面提供了包含N-乙酰基-乳糖胺和/或含有N-乙酰基-乳糖胺的寡糖和益生菌乳杆菌属的物种的制剂。
本发明在第二方面提供了益生菌乳杆菌属的物种和N-乙酰基-乳糖胺和/或含有N-乙酰基-乳糖胺的寡糖在制备预防和/或治疗胃肠道或上呼吸道病原性感染的药物或营养组合物中的用途。
本发明还扩展至在有需要的个体中预防或治疗胃肠道或上呼吸道病原性感染的方法,包括向个体施用治疗量的包含益生菌菌株和益生菌乳杆菌属的物种和N-乙酰基-乳糖胺和/或含有N-乙酰基-乳糖胺的寡糖的制剂。
本发明的发明人先前已证实了从牛乳中获得的N-乙酰化、中性和唾液酸化的寡糖混合物能有效刺激鼠李糖乳杆菌CGMCC 1.3724以减轻细菌毒素诱导的损伤,观察到的保护效应依赖于在OS混合物存在下介导的细菌-宿主细胞的相互作用(crosstalk)(数据未公开)。在不希望受限于理论的情况下,发明人相信N-乙酰基-乳糖胺和/或含有N-乙酰基-乳糖胺的寡糖和益生菌乳杆菌的组合在预防或治疗上呼吸道和胃肠道病原性感染中的效果源于降低病原体粘附腔上皮的能力的协同效应,其通过寡糖对形成束状纤毛粘附和其他粘附(例如紧密粘附素(intimin))的直接效应和刺激的宿主-益生菌乳杆菌的相互作用的间接效应来发挥作用。
发明详述
在本说明书中,当阅读和解释说明书内容、实施例和权利要求时,应当考虑下列名词具有以下定义:
“婴儿”表示年龄低于12个月的儿童;
“婴儿配方食品”表示预期用来提供生命前四到六个月的婴儿完整营养的食物,和直到12个月为止作为其他食物的补充物;
“益生菌”表示对宿主的健康或良好状态具有有益效果的微生物细胞制剂或微生物细胞组分(Salminen S,Ouwehand A.Benno Y.等“probiotics:how should they be defined”Trend Food Sci.Technol.1999:10107-10);
本发明涉及N-乙酰基-乳糖胺和/或含有N-乙酰基-乳糖胺的寡糖。含有N-乙酰基-乳糖胺的合适的寡糖包括乳糖-N-四糖(LNT)和乳糖-N-新四糖(LNnT)。LNT和LNnT可通过使用糖基转移酶从供体部分经酶促转移糖单元至受体部分来化学合成,例如US专利5,288,637中所述。或者,LNT和LNnT可通过游离或结合于寡糖(例如乳果糖)的酮-己糖(例如果糖)的化学转化形成N-乙酰基己糖胺或含有N-乙酰基己糖胺的寡糖进行制备,如Wrodnigg,T.M.;Stutz,A.E.(1999)Angew.Chem.Int.Ed.38:827-828所述。通过该方法生产的N-乙酰基乳糖胺随后可转运至作为受体部分的乳糖。
益生菌乳杆菌可选自符合益生菌定义的任何乳杆菌菌株,其具有掺入本发明的制剂中的产品的可接受的贮存期限。例如,婴儿配方食品需要在直到36个月之内保持稳定和有效。当然,本发明的制剂并不需要整合到其他产品例如食品中,但可作为食品或与适当的赋形剂混合,以例如粉末或胶囊或压缩片剂的形式摄入。
优选的乳杆菌属的物种实例是鼠李糖乳杆菌、副干酪乳杆菌(Lactobacillus paracasei)和罗伊乳杆菌(Lactobacillus reuteri)。特别优选的菌株是鼠李糖乳杆菌ATCC 53103、鼠李糖乳杆菌CGMCC 1.3724、副干酪乳杆菌CNCM I-2116、罗伊乳杆菌ATCC 55730和罗伊乳杆菌DSM17938。
可以根据任何本领域已知的合适方法培养选定的益生菌乳杆菌,并制备以用于添加到本发明的制剂或营养组合物中,例如通过冷冻干燥或喷雾干燥。或者,菌株可从专业供应商例如Christian Hansen和Morinaga处购买,其已经被制备成用于向营养组合物如婴儿配方食品中添加的合适形式。
本发明的制剂可提供每g寡糖为102到1010cfu的益生菌。
在本发明一个优选的方面,上述制剂添加到营养组合物中。在本发明的上下文中,术语“营养组合物”包括任何可消费的产品。因此,它可以是用于人类消费的产品,尤其是婴儿配方食品、成人乳等。然而,所述制剂的消费不限于婴儿和儿童,特别的,本发明的制剂可掺入到脱水乳或谷类混合物中。
营养组合物优选是婴儿配方食品,其含有0.1到3g/100g组合物(基于干重)的N-乙酰基-乳糖胺和/或含有N-乙酰基-乳糖胺的寡糖。
本发明的婴儿配方食品可以含有不超过2.0g/100kcal、优选1.8-2.0g/100kcal的蛋白质源。认为蛋白质类型对于本发明并不关键,只要满足必需氨基酸的最小需求量并且能够保证满意的生长即可,但是优选其中超过50重量%的蛋白质源是乳清。所以,可以使用基于乳清、酪蛋白及其混合物的蛋白质源,以及基于大豆的蛋白质源。就乳清蛋白质来说,蛋白质源可以基于酸乳清或甜乳清或其混合物,可包括以任何所需比例的α-乳球蛋白和β-乳球蛋白。
蛋白质可以是完整的或水解的或完整和水解蛋白质的混合物。可能需要提供部分水解的蛋白质(水解度2-20%),例如对于认为有发生牛奶过敏的婴儿。如果需要水解的蛋白质,可以根据需要和本领域已知的情况进行水解过程。例如,乳清蛋白质水解物可以通过在一个或多个步骤中酶水解乳清组分来制备。如果用作起始材料的乳清组分基本不含乳糖,那么发现蛋白质在水解过程中会遇到更少的赖氨酸阻断(lysine blockage)。这使得赖氨酸阻断的程度从总赖氨酸的约15重量%降低到少于10重量%;例如约7重量%的酪氨酸,这大大的提高了蛋白质源的营养质量。
婴儿配方食品可含有碳水化合物源。可以使用任何常规用于婴儿配方食品的碳水化合物源,例如乳糖、蔗糖、麦芽糖糊精、淀粉及其混合物,但优选的碳水化合物源是乳糖。优选的,该碳水化合物源占该配方食品的总能量的35-65%。
婴儿配方食品也可含有脂类源。该脂类源可以是适用于婴儿配方食品的任何脂类或脂肪。优选的脂肪源包括棕榈酸油酸甘油酯、高油酸葵花籽油和高油酸红花油。例如可以加入含有大量预先形成的花生四烯酸和二十二碳六烯酸的少量油类,例如鱼油或微生物油一样,也可以加入必需脂肪酸亚油酸和α-亚麻酸。总的来说,优选脂肪含量为例如占该配方食品的总能量的30-55%。脂肪源优选n-6脂肪酸与n-3脂肪酸的比例为约5∶1至约15∶1;例如约8∶1至约10∶1。
婴儿配方食品也可含有营养有效量的被认为对日常饮食必须的所有维生素和矿物质。已经确定了对某些维生素和矿物质的最低需求。任选存在于婴儿配方食品中的矿物质、维生素和其他营养物质的实例包括维生素A、维生素B1、维生素B2、维生素B6、维生素B12、维生素E、维生素K、维生素C、维生素D、叶酸、肌醇、尼克酸、生物素、泛酸、胆碱、钙、磷、碘、铁、镁、铜、锌、锰、氯、钾、钠、硒、铬、钼、牛磺酸和L-肉碱。矿物质通常以盐的形式加入。特定矿物质和其他维生素的存在和量根据所针对的婴儿群体而改变。
在需要时,婴儿配方食品可含有乳化剂和稳定剂,例如大豆卵磷脂、柠檬酸甘油单酯和甘油二酯等。
除所述N-乙酰基-乳糖胺和/或含有N-乙酰基-乳糖胺的寡糖外,所述婴儿配方食品优选还包含0.3-10%的量的至少一种益生元。益生元是不消化的食品成分,其通过选择性的刺激结肠中一种或有限数量的细菌的生长和/或活性来有益的影响宿主,从而增进宿主健康。此类成分不消化是因为其在胃或小肠中不能被分解和吸收,从而可以完整的通过并到达结肠,在这里由有益细菌选择性地进行发酵。益生元的实例包括某些寡糖,例如寡聚果糖(FOS)和半乳寡聚糖(GOS)。可以使用组合的益生元,例如90%GOS和10%短链寡聚果糖(例如以商品名Raftilose出售的产品)或10%的菊糖(例如以商品名Raftiline出售的产品)。尤其优选的益生元组合是包含以下成分的混合物:5-70重量%的至少一种N-乙酰化寡糖,其选自GalNAcα1,3Galβ1,4Glc和Galβ1,6GalNAcα1,3Galβ1,4Glc;20-95重量%的至少一种中性寡糖,其选自Galβ1,6Gal、Galβ1,6Galβ1,4Glc、Galβ1,6Galβ1,6Glc、Galβ1,3Galβ1,3Glc、Galβ1,3Galβ1,4Glc、Galβ1,6Galβ1,6Galβ1,4Glc、Galβ1,6Galβ1,3Galβ1,4Glc、Galβ1,3Galβ1,6Galβ1,4Glc和Galβ1,3Galβ1,3Galβ1,4Glc;以及2-50重量%的至少一种唾液酸化的寡糖,其选自NeuAcα2,3Galβ1,4Glc和NeuAcα2,6Galβ1,4Glc。该混合物可从动物乳中制备,例如WO2007/101675中所描述。
婴儿配方食品可任选地含有可能具有有益效果的其他物质,例如乳铁蛋白、核苷酸、核苷等。
如果要将本发明的制剂加入婴儿配方食品或其他基于乳的营养组合物,该组合物可以以本领域已知的任何适宜的方式制备。例如,可通过将蛋白质源、任何除乳糖外的碳水化合物和脂肪源以合适的比例混合制备婴儿配方食品。在需要时可加入乳化剂。此时可加入维生素和矿物质,但是通常稍后加入,以避免热降解。任何亲脂性维生素、乳化剂等可在混合前溶解在脂肪源中。然后可将水,优选经过反渗透处理的水加入混合,以形成液体混合物。
随后可对所述液体混合物热处理以减少细菌载量。例如可快速加热液体混合物到约80℃至约110℃的范围内约5秒至约5分钟。这可以通过蒸汽注入或热交换器进行,例如平板热交换器。
然后将液体混合物冷却至约60℃至约85℃,例如通过瞬时冷却。然后将液体混合物匀浆,例如在两个阶段中匀浆,第一阶段大约7MPa至约40MPa,第二阶段大约2MPa至约14MPa。均质化的混合物可进一步冷却,以便加入任何热敏感的成分,例如维生素和矿物质。此时可以方便的标准化该均质混合物的pH和固体含量。
将均质化的混合物转移至适宜的干燥装置中,例如喷雾干燥器或冷冻干燥机中,变成粉末。这些粉末应具有低于约5重量%的含水量。
本发明的制剂可预先制备,并通过干法混合直接加到营养组合物中。或者,益生菌乳杆菌和N-乙酰基-乳糖胺和/或含有N-乙酰基-乳糖胺的寡糖可通过干法混合分别加入到营养组合物中。
在另一个实施方案中,本发明的制剂可以是补充物的形式,其包括足以获得个体所需效应的量的益生菌乳杆菌和N-乙酰基-乳糖胺和/或含有N-乙酰基-乳糖胺的寡糖。优选的,N-乙酰基-乳糖胺和/或含有N-乙酰基-乳糖胺的寡糖的日剂量为0.1-3g,益生菌乳杆菌的日剂量为10e5-10e12cfu。补充物中所包含的N-乙酰基-乳糖胺和/或含有N-乙酰基-乳糖胺的寡糖和益生菌乳杆菌的量将根据如何施用所述补充物而相应地进行选择。例如,如果一天两次施用补充物,则每次补充物可含有0.05-1.5g N-乙酰基-乳糖胺和/或含有N-乙酰基-乳糖胺的寡糖和10e3-10e6cfu的益生菌乳杆菌。补充物可以是例如片剂、胶囊、软锭剂或液体形式。所述补充物还可包含保护性水胶体(例如树胶、蛋白质、改性淀粉)、粘合剂、薄膜成型剂、包封剂/材料、壁/壳材料、基质化合物、包衣、乳化剂、表面活性剂、增溶剂(油、脂肪、蜡类、卵磷脂等)、吸附剂、载体、填充剂、辅助化合物、分散剂、湿润剂、操作助剂(溶剂)、流动剂、味觉掩蔽剂、填充剂、胶凝剂和凝胶成型剂。补充物也可以含有常规的药用添加剂和佐剂、赋形剂和稀释剂,包括但不限于水、任何来源的明胶、植物胶、木素磺酸盐、滑石、糖、淀粉、阿拉伯胶、植物油、聚亚烷基二醇、矫味剂、防腐剂、稳定剂、乳化剂、缓冲液、润滑剂、着色剂、湿润剂、填充剂等。
此外,所述补充物可含有适合于口服或肠内施用的有机或无机载体物质,以及维生素、矿物质痕量元素和符合政府规章推荐例如USRDA的其他微量营养素。
本发明的制剂可有效预防和治疗上呼吸道和胃肠道的细菌和病毒的病原性感染,包括轮状病毒和诺沃克样病毒、肠致病性大肠杆菌(EPEC)、肠出血性大肠杆菌(EHEC)、肠毒性大肠杆菌(ETEC)和沙门氏菌属的物种(Salmonella sp)所引起的胃肠道感染,和包括由呼吸道合胞病毒、博卡病毒(bocaviruses)、鼻病毒属、肺炎链球菌、流感嗜血菌和粘膜炎莫拉菌所引起的上呼吸道感染。本发明的制剂可以特别有效地预防和治疗中耳炎,这是一种与经常发生的呼吸道感染相关的病症。
现在通过参考下列实施例来进一步说明本发明。
实施例1
下表列出了适用于本发明的婴儿配方食品的组成的实例:
营养成分 | 每100kcal | 每升 |
能量(kcal) | 100 | 670 |
蛋白质(g) | 1.83 | 12.3 |
脂肪(g) | 5.3 | 35.7 |
亚油酸(g) | 0.79 | 5.3 |
α-亚麻酸(mg) | 101 | 675 |
乳糖(g) | 11.2 | 74.7 |
矿物质(g) | 0.37 | 2.5 |
Na(mg) | 23 | 150 |
K(mg) | 89 | 590 |
Cl(mg) | 64 | 430 |
Ca(mg) | 62 | 410 |
P(mg) | 31 | 210 |
Mg(mg) | 7 | 50 |
Mn(μg) | 8 | 50 |
Se(μg) | 2 | 13 |
维生素A(μg RE) | 105 | 700 |
维生素D(μg) | 1.5 | 10 |
维生素E(mg TE) | 0.8 | 5.4 |
Claims (10)
1.包含N-乙酰基-乳糖胺和/或含有N-乙酰基-乳糖胺的寡糖和益生菌乳杆菌属的物种的制剂。
2.权利要求1的制剂,其中含有N-乙酰基-乳糖胺的寡糖是乳糖-N-四糖和乳糖-N-新四糖。
3.权利要求1或2的制剂,其中益生菌乳杆菌属的物种是鼠李糖乳杆菌ATCC 53103、鼠李糖乳杆菌CGMCC 1.3724、罗伊乳杆菌ATCC 55730或罗伊乳杆菌DSM 17938。
4.包含上述权利要求任一项所述制剂的营养组合物。
5.权利要求4的营养组合物,其是婴儿配方食品。
6.权利要求4或5的食物产品,其包含基于干物质重量的0.3-4%的权利要求1-3任一项所述的制剂。
7.权利要求1-3任一项所述的制剂在制备预防或治疗胃肠道病原性感染的营养组合物或药物中的用途。
8.权利要求1-3任一项所述的制剂在制备预防或治疗上呼吸道病原性感染的营养组合物或药物中的用途。
9.权利要求1-3任一项所述的制剂在制备预防或治疗中耳炎的营养组合物或药物中的用途。
10.权利要求7、8或9的用途,其中所述感染由致病菌引起。
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CN104968216A (zh) * | 2012-12-10 | 2015-10-07 | N·V·努特里奇亚 | 含有非消化性寡糖的营养组合物 |
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CN104203008A (zh) * | 2012-04-10 | 2014-12-10 | Cj第一制糖株式会社 | 含有塔格糖和益生乳酸菌的合益素食品组合物 |
CN104968216A (zh) * | 2012-12-10 | 2015-10-07 | N·V·努特里奇亚 | 含有非消化性寡糖的营养组合物 |
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US9254303B2 (en) | 2016-02-09 |
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