CN107949396A - 包含水解酪蛋白的营养组合物 - Google Patents
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Abstract
通过给予受试者包含广泛水解酪蛋白、广泛水解酪蛋白级分或其组合的营养组合物而调节激酶的方法。受调节的激酶可以是调节炎性信号传导、免疫耐受、代谢信号传导、细胞周期和生长因子信号传导的激酶。营养组合物可以剂量依赖性地抑制一系列丝氨酸、苏氨酸和酪氨酸激酶。
Description
技术领域
本公开内容涉及通过给予受试者广泛水解酪蛋白和/或其级分(“eHC”)来调节特定的激酶活性的方法。eHC可抑制一系列丝氨酸、苏氨酸和酪氨酸激酶。受试者可以是人类,特别是人类婴儿或儿童。
背景
激酶是催化磷酸化(磷酸基团转移)的一类酶。激酶调节许多复杂的过程,并且可以调节例如炎性信号传导、免疫耐受、代谢信号传导、细胞周期和生长因子信号传导以及在例如炎性信号传导、免疫耐受、代谢信号传导、细胞周期和生长因子信号传导中起重要信号传导作用。因此,抑制激酶可减少或预防炎症、增加免疫耐受性,并有益于代谢信号传导、细胞周期和生长因子信号传导。
因此,提供调节激酶以减少或预防炎症、增加免疫耐受性、并对代谢信号传导、细胞周期和生长因子信号传导有益的方法将是有益的。该方法可包括将营养组合物给予受试者,特别是婴儿或儿童。此外,该方法应该在动物,特别是人类婴儿中在功能上良好耐受,并且不应产生或引起过多的气体、腹胀、胃气胀或腹泻。
发明内容
简言之,在一个实施方案中,本公开内容涉及通过给予受试者包含广泛水解酪蛋白、广泛水解酪蛋白级分或其组合的营养组合物来调节一种或多种激酶的方法。
在一个实施方案中,调节是炎性信号传导激酶的调节。炎性信号传导激酶可以是IKKβ, IRAK4, ITK, JAK1, JAK3, JNK1 (MAPK8), JNK2, JNK3 (MAPK10), LCK,MAPKAPK2, p38α, SYK, COT (MAP3k8), FYN同种型A, FYN同种型B, KIT, MAP3k2,SPHK1, SPHK2, FMS, BTK, Erk1 (MAPK3), Erk2 (MAPk1), Erk5 (MAPk7)或其一种或多种的组合。
在另一个实施方案中,调节是代谢信号传导激酶的调节。代谢信号传导激酶可以是AKT1, AMPKα1/β1/γ1, p70S6K, PDK1, Erk2, SGK或其一种或多种的组合。
在又一个实施方案中,调节是细胞周期激酶的调节。细胞周期激酶可以是AurA,CDK2/CycA2复合物, CHK1或其一种或多种的组合。
在其它实施方案中,调节的激酶是生长因子信号传导激酶。调节的激酶可以是IGF1R, MET, PDGFRα, EGFR, EPHA2, EPHB4, FGFR1, FLT3, GSK3β, HGK, KDR, ABL,SRC, TIE2, TRKA, TYRO3的一种,或其一种或多种的组合。
所述方法还可以是调节另外的激酶,例如CAMK4, CK1ε, CSK, DAPK1, DYRK1B,MST1, NEK2, PAK2, PBK, PIm1, PKACα, PKCα, PKD2, PYK2, ROCK1, TSSK1或其一种或多种的组合。
所公开的方法中的营养组合物可以是婴儿配方,并且在一些实施方案中可以进一步包含脂肪、碳水化合物、益生菌、益生元或其组合。益生元可以包括聚葡萄糖和/或寡聚半乳糖。
实施本发明的最佳方式
现在将详细地提及本公开内容的实施方案,下文阐述了其一个或多个实例。各个实例通过解释本公开内容的营养组合物来提供,并且不是限制。实际上,对于本领域技术人员将是显而易见的是,可在不偏离本公开内容的范围的情况下对本公开内容的教导进行各种修改和变动。例如,作为一个实施方案的一部分说明或描述的特征,可以与另一个实施方案一起使用以产生又一个实施方案。
因此,需要的是,本公开内容涵盖落入随附权利要求书及其等同内容的范围的这类修改和变动。本公开内容的其它目的、特征和方面公开于下面的详细描述中,或从下面的详细描述来看是显而易见的。本领域普通技术人员要了解,本论述只是示例性实施方案的描述,并无意限制本公开内容的更宽泛的方面。
本公开内容总体上涉及包括适合给予儿科受试者的营养组合物的方法。
“营养组合物”意指满足受试者的营养需要的至少一部分的物质或制剂。术语“营养品”、“营养配方”、“肠内营养品”和“营养补剂”在整个公开内容中用作营养组合物的非限制性实例。而且,“营养组合物”可指液体剂、粉剂、凝胶剂、糊剂、固体剂、浓缩剂、混悬剂或即用形式的肠内配方、口服配方、婴儿配方、儿科受试者配方、儿童配方、成长乳和/或成人配方。
术语“肠内”意指可通过胃肠道或消化道或在胃肠道或消化道内传递的。“肠内给予”包括经口喂食、胃内喂食、经幽门给予或任何其它进入消化道的给予。“给予”比“肠内给予”更广义并包括胃肠外给予或使物质吸收进入受试者体内的任何其它给予途径。
“儿科受试者”意指年龄不足13岁的人。在一些实施方案中,儿科受试者是指介于出生和8岁之间的人类受试者。在其它实施方案中,儿科受试者是指介于1到6岁之间的人类受试者。在又一个的实施方案中,儿科受试者是指介于6到12岁之间的人类受试者。术语“儿科受试者”可指如下所述的婴儿(早产或足月)和/或儿童。
“婴儿”意指年龄范围从出生至不满1岁的人类受试者并包括从0至12个月的矫正年龄的婴儿。短语“矫正年龄”意指婴儿的实足年龄减去婴儿早产的时间的量。因此,如果怀胎已达足月,则矫正年龄是该婴儿的年龄。术语婴儿包括低出生体重婴儿、非常低出生体重婴儿、极低出生体重婴儿和早产婴儿。“早产儿”意指在妊娠的第37周结束前出生的婴儿。“晚期早产儿”是指妊娠的第34周和第36周之间的婴儿。“足月”意指在妊娠的第37周结束后出生的婴儿。“低出生体重婴儿”意指出生体重少于2500克(大约5磅,8盎司)的婴儿。“非常低出生体重婴儿”意指出生体重少于1500克(大约3磅,4盎司)的婴儿。“极低出生体重婴儿”意指出生体重少于1000克(大约2磅,3盎司)的婴儿。
“儿童”意指年龄范围从12个月至13岁的受试者。在一些实施方案中,儿童是年龄介于1和12岁之间的受试者。在其它实施方案中,术语“儿童(children / child)”是指介于1和约6岁之间或介于约7和约12岁之间的受试者。在其它实施方案中,术语“儿童”是指介于12个月和约13岁之间的任何年龄范围。
“儿童的营养制品”指满足儿童的至少一部分营养需要的组合物。成长乳是儿童的营养制品的实例。
术语“水解度”指肽键被水解方法破坏的程度。蛋白质水解度(为了表征营养组合物的水解蛋白组分的目的)由制剂领域普通技术人员通过定量所选择的制剂的蛋白质组分的氨基氮与总氮比例(AN/TN)容易地测定。氨基氮组分通过用于测定氨基氮含量的USP滴定方法定量,而总氮组分通过Kjeldahl方法测定,所有这些都是对于分析化学领域的普通技术人员而言熟知的方法。
当蛋白质的肽键通过酶促水解破坏时,每个肽键破坏释出一个氨基,引起氨基氮的增加。应该注意到,甚至未水解蛋白会含有一些暴露的氨基。水解蛋白也会具有与未水解蛋白(从该未水解蛋白形成水解蛋白)不同的分子量分布。水解蛋白的功能和营养性质可受不同大小的肽影响。分子量概况通常通过列出特定范围的分子量(以道尔顿计)级分(例如,2,000-5,000道尔顿,大于5,000道尔顿)的重量百分比给出。
当涉及水解的蛋白质或蛋白质水解产物时,术语“摩尔质量分布”与蛋白质水解产物中存在的每种肽的摩尔质量有关。例如,具有大于500道尔顿的摩尔质量分布的蛋白质水解产物是指包含在蛋白质水解产物中的每种肽具有至少500道尔顿的摩尔质量。为了生产具有大于500道尔顿的摩尔质量分布的蛋白质水解产物,蛋白质水解产物可以进行某些过滤程序或本领域已知的用于除去摩尔质量小于500道尔顿的肽、氨基酸和/或其它蛋白质性材料的任何其它程序。为了本公开内容的目的,可以使用本领域已知的任何方法来生产摩尔质量分布大于500道尔顿的蛋白质水解产物。
术语“蛋白质等同物”或“蛋白质等同物源”包括任何蛋白质源,例如大豆、蛋、乳清或酪蛋白,以及非蛋白质源,例如肽或氨基酸。此外,蛋白质等同物源可以是本领域中使用的任何物质,例如脱脂乳、乳清蛋白、酪蛋白、大豆蛋白、水解蛋白、氨基酸等。可用于实施本公开内容的牛乳蛋白源包括但不限于乳蛋白粉、乳蛋白浓缩物、乳蛋白分离物、脱脂乳固体、脱脂乳、脱脂奶粉、乳清蛋白、乳清蛋白分离物、乳清蛋白浓缩物、甜乳清、酸乳清、酪蛋白、酸性酪蛋白、酪蛋白酸盐(例如酪蛋白酸钠、酪蛋白酸钠钙、酪蛋白酸钙)、大豆蛋白及其任何组合。在一些实施方案中,蛋白质等同物源可以包含水解蛋白质,包括部分水解蛋白和广泛水解蛋白。在一些实施方案中,蛋白质等同物源可以包括完整蛋白质。
术语“蛋白质等同物源”也包括游离氨基酸。在一些实施方案中,氨基酸可以包括但不限于组氨酸、异亮氨酸、亮氨酸、赖氨酸、甲硫氨酸、半胱氨酸、苯丙氨酸、酪氨酸、苏氨酸、色氨酸、缬氨酸、丙氨酸、精氨酸、天冬酰胺、天冬氨酸、谷氨酸、谷氨酰胺、甘氨酸、脯氨酸、丝氨酸、肉毒碱、牛磺酸及其混合物。在一些实施方案中,氨基酸可以是支链氨基酸。在某些其它实施方案中,可以包含小氨基酸肽作为营养组合物的蛋白质组分。这样的小氨基酸肽可以是天然存在的或合成的。
术语“部分水解的”意指具有大于0%但少于约50%的水解度。
术语“广泛水解的”意指具有大于或等于约50%的水解度。因此,“广泛水解酪蛋白级分”是指具有大于或等于约50%的水解度的酪蛋白。在一些实施方案中,广泛水解可以包括大于约80%的水解度。在进一步的实施方案中,广泛水解可以包括大于约90%的水解度。“eHC”是指广泛水解酪蛋白和/或其级分。
术语“无蛋白的”意指含不可测量的量的完整蛋白,如通过标准蛋白检测方法如十二烷基(月桂基)硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)或尺寸排阻色谱法测量的。在一些实施方案中,营养组合物基本上不含蛋白,其中“基本上不含”如下文所定义。
“婴儿配方”意指满足婴儿的至少部分营养需求的组合物。在美国,21 C.F.R.第100、106和107章阐述的联邦法规规定了婴儿配方的内含物。这些法规限定尽力模拟人母乳的营养和其它性质的大量营养物、维生素、矿物质和其它成分水平。
术语“成长乳”是指一大类营养组合物,其预期用作多样化膳食的一部分,以支持年龄介于约1和约6岁之间的儿童的正常成长和发育。
“乳-基”指包含至少一种已经从哺乳动物的乳腺中吸出或提取的组分。在一些实施方案中,乳-基营养组合物包含源自驯养的有蹄类动物、反刍动物或其它哺乳动物的乳的组分或其任何组合。此外,在一些实施方案中,乳-基意指包含牛酪蛋白、乳清、乳糖或其任何组合。而且,“乳-基营养组合物”可指包含本领域已知的任何源自乳的或乳-基产品的任何组合物。
“乳”指一种已经从哺乳动物的乳腺中吸出或提取的组分。在一些实施方案中,营养组合物包含源自驯养的有蹄类动物、反刍动物或其它哺乳动物的乳的组分或其任何组合。
“分级分离(fractionation)过程”包括其中一定量的混合物被分成许多较小量(称为级分(fraction))的任何过程。所述级分可以在组成上不同于所述混合物和其它级分两者。分级分离过程的实例包括但不限于,熔融分级分离(melt fractionation)、溶剂分级分离、超临界流体分级分离和/或其组合。
“脂肪球”指一种由磷脂和其它膜和/或血清蛋白环绕的小质量脂肪,其中脂肪本身可以是任何植物或动物脂肪的组合。
“营养完全的”意指可用作营养的唯一来源的组合物,其可供应基本上所有每日必需量的维生素、矿物质和/或痕量元素与蛋白质、碳水化合物和脂质的组合。实际上,“营养完全的”描述了提供支持受试者的正常生长和发育所需的足量碳水化合物、脂质、必需脂肪酸、蛋白质、必需氨基酸、条件必需氨基酸、维生素、矿物质和能量的营养组合物。
因此,按定义,对早产婴儿是“营养完全的”营养组合物会提供早产婴儿生长所需要的质量上和数量上足量的碳水化合物、脂质、必需脂肪酸、蛋白质、必需氨基酸、条件必需氨基酸、维生素、矿物质和能量。
按定义,对足月婴儿是“营养完全的”营养组合物会提供足月婴儿生长所需要的质量上和数量上足量的所有碳水化合物、脂质、必需脂肪酸、蛋白质、必需氨基酸、条件必需氨基酸、维生素、矿物质和能量。
按定义,对儿童是“营养完全的”营养组合物会提供儿童生长所需要的质量上和数量上足量的所有碳水化合物、脂质、必需脂肪酸、蛋白质、必需氨基酸、条件必需氨基酸、维生素、矿物质和能量。
当应用于营养物时,术语“必需的”是指机体不能以足以用于正常生长和维持健康的量合成因此必须通过膳食供应的任何营养物。术语“条件必需的”在应用于营养物时意指在一定条件下必须通过膳食供应营养物,所述条件是机体无法获得足量的前体化合物用于发生内源合成。
“益生菌”意指对宿主的健康发挥有益作用的具有低致病性或无致病性的微生物。
术语“灭活的益生菌”意指这样的益生菌,其中所提及的益生菌的代谢活性或繁殖能力已经减少或被破坏。然而,“灭活的益生菌”在细胞水平上仍然保留其细胞结构或其它与细胞有关的结构,例如表多糖和至少一部分其生物二醇-蛋白和DNA/RNA结构。如本文所用的,术语“灭活的”与“无活力的”同义。
“益生元”意指不易消化的食物成分,其通过选择性地刺激消化道中一种或有限数量的可改善宿主健康的细菌的生长和/或活动而有益地影响宿主。
“支链脂肪酸” (“BCFA”)意指包含从碳链分支的碳成分的脂肪酸。典型地,分支是烷基分支,特别是甲基,但乙基和丙基分支也是已知的。与同等的直链脂肪酸比较,加入甲基分支会降低熔点。这包括在碳链上具有偶数个碳原子的支链脂肪酸。这些例子可以是十四烷酸、十六烷酸的异构体。
“奇数支链脂肪酸” (“OBCFA”)是具有奇数个碳原子且在碳链上具有一个或多个烷基分支的BCFA的子集。牛奶中发现的主要奇数支链脂肪酸包括,但不限于,十四烷酸、十五烷酸、十六烷酸和十七烷酸的异构体。为了本公开内容的目的,术语“BCFA”包括支链脂肪酸和奇数支链脂肪酸二者。
“反式-脂肪酸”意指具有反式-异构体的不饱和脂肪。反式-脂肪可以是单不饱和或多不饱和的。反式指连接于双键包含的碳原子的两个氢原子的排列。在反式排列中,氢在所述键的对侧。因此反式-脂肪酸是在反式几何构型中含有一个或多个双键的脂质分子。
“磷脂”意指含有甘油二酯、磷酸酯基团和简单的有机分子的有机分子。磷脂的例子包括但不限于,磷脂酸、磷脂酰乙醇胺、磷脂酰胆碱、磷脂酰丝氨酸、磷脂酰肌醇、磷脂酰肌醇磷酸、磷脂酰肌醇二磷酸和磷脂酰肌醇三磷酸、神经酰胺磷酸胆碱、神经酰胺磷酰乙醇胺和神经酰胺磷酰甘油。该定义还包鞘磷脂、糖脂和神经节苷脂。
“植物营养素”意指天然存在于植物中的化学化合物。植物营养素可以包括在任何源自植物的物质或提取物中。术语“植物营养素”包含由植物产生的几大类化合物,例如,多酚化合物、花青苷、原花青素和黄烷-3-醇(即儿茶素、表儿茶素),并可以源自,例如,果实、种子或茶提取物。此外,术语植物营养素包括所有的类胡萝卜素、植物甾醇、硫醇和其它源自植物的化合物。此外,技术人员会理解,除了蛋白质、纤维或其它源自植物的组分外,植物提取物还可包括植物营养素,例如多酚类。因此,例如,除了其它源自植物的物质外,苹果或葡萄籽提取物还可包括有益的植物营养素组分,例如多酚类。
“β-葡聚糖”意指所有的β-葡聚糖,包括特殊类型的β-葡聚糖,例如β-1,3-葡聚糖或β-1,3;1,6-葡聚糖。此外,β-1,3;1,6-葡聚糖是一种类型的β-1,3-葡聚糖。因此,术语“β-1,3-葡聚糖”包括β-1,3;1,6-葡聚糖。
“果胶”意指任何天然-存在的寡糖或多糖,其包含可在植物的细胞壁中发现的半乳糖醛酸。具有各种物理和化学性质的不同种类和等级的果胶是本领域已知的。事实上,果胶的结构可在植物之间、组织之间和甚至在单细胞壁内显著地变化。一般来说,果胶由带负电荷的酸性糖(半乳糖醛酸)组成,并且一些酸性基团呈甲酯基团的形式。果胶的酯化程度是用甲醇酯化的连接于吡喃半乳糖基糖醛酸(galactopyranosyluronic acid)单位的羧基的百分率的量度。
具有酯化程度少于50% (即,少于50%的羧基被甲基化以形成甲酯基团)的果胶被分类为低-酯、低甲氧基或低甲基化(“LM”)果胶,而具有50%或更大的酯化程度(即,超过50%的羧基被甲基化)的那些被分类为高-酯、高甲氧基或高甲基化(“HM”)果胶。极低(“VL”)果胶,一种低甲基化果胶的亚组,具有少于约15%的酯化程度。
如本文所用的,“来自非-人来源的乳铁蛋白”意指由除人母乳之外来源产生或从除人母乳之外来源获得的乳铁蛋白。例如,用于本公开内容的乳铁蛋白包括由遗传修饰的生物体产生的人乳铁蛋白以及非人乳铁蛋白。如本文所用的术语“生物体”,指任何连续的生命系统,如动物、植物、真菌或微生物。
如本文所用的,“非人乳铁蛋白”意指具有不同于人乳铁蛋白的氨基酸序列的氨基酸序列的乳铁蛋白。
“病原体”意指引起疾病状态或病理学综合征的生物体。病原体的实例可包括细菌、病毒、寄生虫、真菌、微生物或其组合。
“调节”或“调整”意指使产生修饰、控制和/或调节影响。在一些实施方案中,术语“调节”意指显示对特定组分的水平/量的增加或刺激作用。在其它实施方案中,“调节”意指显示对特定组分的水平/量的减少或抑制作用。
如本文所用的所有百分比、份数和比例均基于总制剂的重量计,除非另外指明。
“每日”给予的所有指定的量可以在24小时周期的过程内给予的1个单位剂量、单份量或二个或更多个剂量或份量递送。
本公开内容的营养组合物可基本上不含任何本文所述的任选的或选择的成分,条件是其余的营养组合物仍含有本文所述的所有必需成分或特征。在本上下文中,并且除非另外指明,否则术语“基本上不含”意指所选的组合物可含有小于功能量的任选成分,通常小于0.1%重量,并且还包括0%重量的所述任选或选择的成分。
本公开内容对单数特征或限制的所有提及应包括相应的复数特征或限制,并且反之亦然,除非在作出提及的上下文中另外指明或明确暗示与之相反。
可以任何顺序进行本文所采用的方法或过程步骤的所有组合,除非在作出提及的组合的上下文中另有指明或明确暗示与之相反。
本公开内容的方法和组合物(包括其组分),可包含本文所述实施方案的必需要素和限制以及本文或别处描述的可用于营养组合物的任何额外或任选的成分、组分或限制;由本文所述实施方案的必需要素和限制以及本文或别处描述的可用于营养组合物的任何额外或任选的成分、组分或限制组成;或基本由它们组成。
本文所用的术语“约”应解释为是指任何范围的端点所指定的两个数字。对范围的任何提及应视为对该范围内的任何子集提供依据。
本公开内容要解决的问题之一涉及激酶活性,并且通过提供一种通过给予受试者广泛水解酪蛋白、广泛水解酪蛋白级分或其组合来调节或调整一种或多种激酶的方法来解决。广泛水解酪蛋白可以在营养组合物,特别是婴儿配方中给予受试者。
eHC剂量依赖性地抑制一系列丝氨酸、苏氨酸和酪氨酸激酶。相反,氨基酸制剂没有显示出相似的活性,这支持了激酶抑制活性可能由eHC衍生的肽介导的论点。
受影响的激酶与例如免疫耐受性和炎症相关。可以将广泛水解酪蛋白加入到营养组合物例如婴儿配方中以降低受试者的炎症。
单独或作为一组,酪氨酸、丝氨酸和苏氨酸激酶可有助于免疫调节,导致获得耐受性。例如,先前已经证实SYK的阻断可以在已建立变态反应的小鼠中预防变应原致敏或诱导脱敏。本文中鉴定的激酶的描述可以在UniProt (Universal Protein Resource,可访问uniprot.org)中找到。
受调节的激酶可以是炎性信号传导激酶,包括KKβ, IRAK4, ITK, JAK1, JAK3,JNK1 (MAPK8), JNK2, JNK3 (MAPK10), LCK, MAPKAPK2, p38α, SYK, COT (MAP3k8),FYN同种型A, FYN同种型B, KIT, MAP3k2, SPHK1, SPHK2, FMS, BTK, Erk1 (MAPK3),Erk2 (MAPk1), Erk5 (MAPk7)或其组合。
IKKβ是充当IκB激酶的蛋白质亚基的酶,其是参与触发免疫反应的细胞因子激活的细胞内信号传导途径的组分。IKK的活性引起转录因子NFκB的激活,其在进入细胞核后激活涉及免疫反应的各种基因。
IRAK-4 (白细胞介素-1受体相关激酶4)是参与从Toll样受体发出天然免疫反应信号的蛋白激酶。它也支持来自T细胞受体的信号传导。没有IRAK-4的动物对病毒和细菌较敏感,但完全抵抗LPS攻击。JAK3(Janus激酶3)是属于Janus激酶家族的酪氨酸激酶。Janus家族的其它成员包括JAK1、JAK2和TYK2。它们是与细胞因子受体特异性缔合的胞质酪氨酸激酶。因为细胞因子受体蛋白缺乏酶活性,所以它们依赖于JAK以在其配体(例如细胞因子)结合时启动信号传导。
JNK1(MAPK8)(c-Jun N端激酶1/促分裂原活化蛋白激酶8)是参与诸如细胞增殖、分化、迁移、转化和程序性细胞死亡等各种过程的丝氨酸/苏氨酸蛋白激酶。细胞外刺激例如促炎细胞因子或物理应激刺激应激激活的蛋白激酶/c-Jun N端激酶(SAP/JNK)信号传导途径。在该级联中,两种双特异性激酶MAP2K4/MKK4和MAP2K7/MKK7磷酸化并激活JNK1。
JNK2(c-Jun N端激酶2)活性调节几种重要的细胞功能,包括细胞生长、分化、存活和凋亡。多种应激刺激可以激活JNK。
JNK3 (MAPK10)(c-Jun N端激酶3/促分裂原活化蛋白激酶10)是参与诸如神经元增殖、分化、迁移和程序性细胞死亡等各种过程的丝氨酸/苏氨酸蛋白激酶。细胞外刺激例如促炎细胞因子或物理应激刺激应激激活的蛋白激酶/c-Jun N端激酶(SAP/JNK)信号传导途径。
LCK(或淋巴细胞特异性蛋白酪氨酸激酶)是在淋巴细胞中发现的56kDa蛋白质。LCK是一种酪氨酸激酶,其将参与细胞内信号传导的蛋白质的酪氨酸残基磷酸化。LCK是酪氨酸激酶的SRC家族的成员,并参与T淋巴细胞中的白细胞介素2(IL-2)信号传导。
MAPKAPK2 (促分裂原活化蛋白激酶活化蛋白激酶2)参与细胞过程,包括应激、炎性反应、核输出、基因表达和细胞增殖。
p38α促分裂原活化蛋白激酶是一类促分裂原活化蛋白激酶。p38a促分裂原活化蛋白激酶响应于炎性信号而被激活。这类激酶对应激刺激例如细胞因子、紫外线照射、热休克和渗透压休克有响应,并参与细胞分化、细胞凋亡和自噬。
SYK(脾酪氨酸激酶)与ZAP70一起是酪氨酸激酶的SYK家族的成员。虽然SYK和ZAP-70主要在造血细胞中表达,但SYK在多种其它组织中也有表达。SYK和ZAP-70分别在B细胞或T细胞中表达,并分别从B细胞受体或T细胞受体传递信号。
ITK(白细胞介素-2诱导型T细胞激酶)参与适应性免疫反应的调节。ITK调节T细胞和非传统NKT细胞的发育、功能和分化。
JAK1(Janus激酶1)是属于Janus激酶家族的酪氨酸激酶。Janus家族的其它成员包括JAK2、JAK3和TYK2。它们是与细胞因子受体特异性缔合的胞质酪氨酸激酶。因为细胞因子受体蛋白缺乏酶活性,所以它们依赖JAK以在其配体(例如细胞因子)结合时启动信号传导。
JAK3(Janus激酶3)调节细胞生长、发育和分化。JAK3参与血细胞组分的发展(造血)。
对于脂多糖(LPS)诱导的TLR4介导的巨噬细胞中MAPK/ERK途径的激活,COT(MAP3k8)(促分裂原活化蛋白激酶激酶激酶8)是必需的,因此对于在免疫反应期间产生促炎细胞因子TNF -α(TNF)是关键的。参与T辅助细胞分化和T细胞中IFNG表达的调节。通过负调节I型干扰素(IFN)产生参与介导宿主对细菌感染的抗性。在体外,以不依赖IRAK1的方式响应于IL1而激活MAPK/ERK途径,导致IL8和CCL4的上调。
FYN同种型A是一种酪氨酸激酶癌基因家族,在多种细胞类型中表达,其生物学功能是多样的,包括通过T细胞受体的信号传导,调节脑功能以及粘附介导的信号传导。在适当的靶组织中改变Fyn的水平可以导致对自身免疫性疾病更好的治疗。
FYN同种型B是另一种酪氨酸激酶癌基因家族,在各种细胞类型中表达,其生物学功能是多样的,包括通过T细胞受体的信号传导,调节脑功能以及粘附介导的信号传导。在适当的靶组织中改变Fyn的水平可导致对自身免疫性疾病更好的治疗。
KIT(酪氨酸激酶受体KIT)是MGF(肥大细胞生长因子,也被称为干细胞因子)的3型跨膜受体。该基因中的突变与胃肠道间质瘤和肥大细胞疾病有关。
MAP3K2(促分裂原活化蛋白激酶激酶激酶2)是丝氨酸/苏氨酸蛋白激酶家族的成员,该激酶优先激活参与MAP激酶信号传导途径的其它激酶。该激酶已显示直接磷酸化并激活IκB激酶,并因此在NF-κB信号传导途径中起作用。还发现该激酶结合并激活蛋白激酶C相关激酶2,这表明其参与调节的信号传导过程。
SPHK1(鞘氨醇激酶1)催化鞘氨醇的磷酸化以形成鞘氨醇-1-磷酸(S1P),一种具有胞内和胞外功能的脂质介质。在细胞内,S1P调节增殖和存活,而在细胞外,它是细胞表面G蛋白偶联受体的配体。这种蛋白质及其产物S1P在炎性、抗细胞凋亡和免疫过程中重要的TNF-α信号传导和NF-κ-B激活途径中起关键作用。
SPHK2(鞘氨醇激酶2)的功能类似于SPHK1。
FMS(集落刺激因子1受体)是集落刺激因子1(一种控制巨噬细胞产生、分化和功能的细胞因子)的受体。该受体介导这种细胞因子的大部分(如果不是全部)生物效应。配体结合通过寡聚和转磷酸过程激活受体激酶。编码的蛋白质是酪氨酸激酶跨膜受体和酪氨酸蛋白激酶的CSF1/PDGF受体家族的成员。
BTK(布鲁顿无丙种球蛋白血症酪氨酸激酶)是一种非受体酪氨酸激酶,对于B淋巴细胞的发育、分化和信号传导是必不可少的。抗原与B细胞抗原受体(BCR)的结合触发最终导致B细胞活化的信号传导。在质膜上BCR接合和激活后,在几个位点使PLCG2磷酸化,通过钙动员引发下游信号传导途径,然后激活蛋白激酶C(PKC)家族成员。
Erk1(MAPK3)(促分裂原活化蛋白激酶3)和Erk2(MAPK1)。MAPK3/ERK1和MAPK1/ERK2属于丝氨酸/苏氨酸激酶,是在MAPK/ERK级联中发挥重要作用的两种MAPK。它们也参与由活化KIT和KITLG/SCF启动的信号传导级联。根据细胞情况,MAPK/ERK级联通过调节转录、翻译、细胞骨架重排介导多种生物功能,例如细胞生长、粘附、存活和分化。MAPK/ERK级联还通过磷酸化许多转录因子在分化细胞的减数分裂、有丝分裂和分裂后功能的启动和调节中发挥作用。
Erk5(MAPK7)(促分裂原活化蛋白激酶7)是MAP激酶家族的成员。MAP激酶作为多种生物化学信号的整合点起作用,并参与多种细胞过程,例如增殖、分化、转录调节和发育。该激酶被促分裂原活化蛋白激酶激酶5(MAP2K5/MEK5)特异性激活。它参与各种受体分子(包括受体型激酶和G蛋白偶联受体)的下游信号传导过程。响应于细胞外信号,该激酶易位至细胞核,其中它通过磷酸化并激活不同的转录因子而调节基因表达。
该方法可以调节代谢信号传导激酶,包括AKT1, AMPK, p70S6K, PDK1, Erk2,SGK或其组合。
AKT1(v-akt鼠胸腺瘤病毒癌基因同系物1)激酶涉及许多信号传导途径,例如细胞分化、细胞存活、凋亡和神经系统形成和功能。
AMPK参与调节细胞能量,响应低ATP水平,并调节脂肪酸氧化和自噬。AMPK包括AMPKα1、AMPKβ1和AMPKγ1。
p70S6K参与细胞信号转导,包括细胞生长和G1期细胞周期进展。
Erk2,也被称为促分裂原活化蛋白激酶1,是细胞外信号调节激酶,参与细胞过程例如分化、发育、增殖和转录调节。
包括SGK1的SGK(血清和糖皮质激素调节的激酶)是离子通道调节剂,包括活化钾、钠和氯离子通道。
PDK(丙酮酸脱氢酶激酶)。丙酮酸脱氢酶是催化丙酮酸氧化脱羧的线粒体多酶复合物的一部分,并且是负责调节哺乳动物中碳水化合物燃料的稳态的主要酶之一。酶活性受磷酸化/去磷酸化循环调节。通过特定的PDK磷酸化PDH导致失活。PDK包括PDK1、PDK2、PDK3和PDK4。
在一个实施方案中,调节是细胞周期激酶的调节。细胞周期激酶可以是AurA、CDK2/CycA2复合物、CHK1或其组合。
AurA (Aurora A)是有丝分裂丝氨酸/苏氨酸激酶家族的成员。它牵涉有丝分裂和减数分裂期间的重要过程,有丝分裂和减数分裂的适当功能对于健康细胞增殖是不可或缺的。在细胞周期的G2期至M期转变期间,Aurora A被磷酸化激活。
CDK2/CycA2复合物(细胞周期蛋白依赖性激酶2复合物)控制细胞周期,对减数分裂至关重要。CDK2/CycA2复合物在人胚胎干细胞的细胞增殖、细胞死亡和DNA修复中起作用。
CHK1/CHEK1(检验点激酶1)在CAMKL激酶家族中。CHK1调节细胞周期、细胞增殖和细胞死亡。
在其它实施方案中,受调节的激酶是生长因子信号传导激酶。受调节的生长因子信号转导激酶可以是IGF1R, MET, PDGFRα, EGFR, EPHA2, EPHB4, FGFR1, FLT3, GSK3β,HGK, KDR, ABL, SRC, TIE2, TRKA, TYRO3中的一种或其组合。
IGF1R(胰岛素样生长因子1受体)介导细胞生长和细胞存活,并可在癌细胞的存活中起作用。IGF1R介导胰岛素样生长因子1(IGF1)的活性。
MET(肝细胞生长因子受体;酪氨酸蛋白激酶Met)从细胞外基质转导信号并调节细胞增殖、破坏、扩散、形态发生、存活和细胞间连接。MET涉及成人的发育和伤口愈合、器官再生和组织重塑中的信号传导。另外,MET的功能障碍可能是癌症的一个原因。
PDGFRα(血小板衍生的生长因子α)介导包括神经胶质细胞在内的中枢神经系统的发育。PDGFRα作为PDGFA、PDGFB和PDGFC的细胞表面受体起作用,并在胚胎发育、细胞增殖、存活和趋化性的调节中起重要作用。根据情况,促进或抑制细胞增殖和细胞迁移。在骨髓衍生的间充质干细胞的分化中发挥重要作用。正常骨骼发育所需。
EGFR(ErbB-1或表皮生长因子受体)调节细胞分裂和细胞。EGFR激活几个信号传导级联以将细胞外信号转化为适当的细胞应答。EGFR激活至少4个主要的下游信号传导级联,包括RAS-RAF-MEK-ERK、PI3激酶-AKT、PLC-γ-PKC和STATs模块。EGFR也可能激活NF-κ-B信号传导级联。还直接磷酸化其它蛋白,如RGS16,激活其GTP酶活性,并可能将EGF受体信号传导偶联至G蛋白偶联受体信号传导。EGFR可能与癌细胞和神经退行性疾病的发展有关。
EPHA2 (ephrin A型2)调节神经和血管系统的发育。EPHA2与位于相邻细胞上的膜结合的ephrin-A家族配体结合,导致接触依赖的双向信号传导至相邻细胞。EPHA2可参与UV辐射诱导的细胞凋亡,并在若干胎儿组织的发育中发挥作用。参与血管生成。
EPHB4 (ephrin B型4)调节神经系统的发育。EPHB4与位于相邻细胞上的跨膜ephrin-B家族配体结合,导致接触依赖的双向信号传导至相邻细胞。EPHB4通过调节细胞粘附和细胞迁移在心脏形态发生和血管生成中起核心作用。EPHB4介导的正向信号传导控制与表达EFNB2的细胞的细胞排斥和分离。EPHB4参与出生后血管重塑,形态发生和渗透性,因此在肿瘤血管生成的情况下是重要的。
FGFR1 (成纤维细胞生长因子受体1)调节细胞增殖、分化和迁移。FGFR1作为成纤维细胞生长因子的细胞表面受体起作用,与Pfeiffer综合征、Jackson-Weiss综合征、Antley-Bixler综合征和癌症有关。
HGK (肝细胞祖细胞激酶样或生发中心激酶样)调节细胞转化、侵袭和粘附。
KDR (激酶插入结构域受体或血管内皮生长因子受体2)调节血管生成、血管发育和细胞分化。
ABL (酪氨酸-蛋白激酶ABL1;Abelson酪氨酸-蛋白激酶1) ABL在与细胞生长和存活相关的许多关键过程中起作用,例如响应细胞外刺激的细胞骨架重塑,细胞运动性和粘附,受体内吞作用,自噬,DNA损伤反应和细胞凋亡。通过控制细胞骨架动力学的蛋白质的酪氨酸磷酸化协调肌动蛋白重塑,所述蛋白质例如WASF3(参与分支形成);ANXA1(参与膜锚定);DBN1,DBNL,CTTN,RAPH1和ENAH(参与信号传导);或MAPT和PXN(微管结合蛋白)。
SRC (原癌基因酪氨酸-蛋白激酶Src)调节细胞分裂、运动性、粘附和存活。SRC参与控制包括基因转录、免疫反应、细胞粘附、细胞周期进程、凋亡、迁移和转化在内的多种生物学活性的信号传导途径。
TIE2 (促血管生成素-1受体;蛋白受体酪氨酸激酶,上皮特异性的)调节血管生成并作为ANGPT1、ANGPT2和ANGPT4的细胞表面受体起作用和调节血管生成、内皮细胞存活、增殖、迁移、粘附和细胞扩散、肌动蛋白细胞骨架重组,以及血管静止的维持。TIE2通过防止促炎性血浆蛋白和白细胞从血管泄漏而具有抗炎作用。
TRKA(原肌球蛋白受体激酶A,高亲和力神经生长因子受体,神经营养酪氨酸激酶受体1型或TRK1转化酪氨酸激酶蛋白)调节细胞分化和神经系统发育。
TYRO3 (TYRO3蛋白酪氨酸激酶)调节精子发生、免疫调节、吞噬作用和控制细胞存活和增殖。
GSK3β (糖原合酶激酶-3)是最初被鉴定为糖原合酶的磷酸化剂和失活剂的脯氨酸指导的丝氨酸-苏氨酸激酶。GSK3β参与能量代谢、神经元细胞发育和身体模式形成。
所述方法还可调节另外的激酶,例如CAMK4, CK1ε, CSK, DAPK1, DYRK1B, MST1,NEK2, PAK2, PBK, PIM1, PKACα, PKCα, PKD2, PYK2, ROCK1, TSSK1或其组合。
CAMK4(钙/钙调蛋白依赖性蛋白激酶IV型)调节淋巴细胞、神经元和雄性生殖细胞中的转录。
CK1ε(酪蛋白激酶1ε)调节基因转录和翻译。
CSK(c-src酪氨酸激酶)调节细胞生长、分化、迁移和免疫反应。
DAPK1(死亡相关蛋白激酶1)调节细胞凋亡。
DYRK1B (双特异性酪氨酸-(Y)-磷酸化调节激酶1B)调节核细胞功能。
MST1 (巨噬细胞刺激1)调节细胞凋亡、神经系统发育、器官生长和血管模式化。
NEK2(丝氨酸/苏氨酸蛋白激酶Nek2)调节细胞减数分裂。
PAK2(p21活化的激酶)调节细胞凋亡。
PBK (淋巴因子激活的杀伤T细胞来源的蛋白激酶)磷酸化MAP激酶p38,并可在淋巴细胞激活中发挥作用。
PIM1(原癌基因丝氨酸/苏氨酸蛋白激酶)主要在脾、胸腺、骨髓、前列腺、口腔上皮、海马和胎儿肝细胞中表达,PIM1也被发现在自人类肿瘤分离的细胞培养物中高度表达。PIM1主要参与细胞周期进展、凋亡和转录激活,以及更一般的信号转导途径。
PKACα(蛋白激酶A催化亚基α)调节转录。
PKCα(蛋白激酶C-α)调节细胞粘附、细胞转化、细胞周期检验点和细胞体积控制。
PKD2(蛋白激酶D)调节细胞增殖、存活和免疫反应。
PYK2(富含脯氨酸的酪氨酸激酶)调节与血小板和免疫细胞相关的途径。
ROCK1 (Rho相关,含有卷曲螺旋的蛋白激酶1)调节与成纤维细胞、血小板和免疫细胞相关的途径。
TSSK1(睾丸特异性丝氨酸激酶1)调节与精子发生相关的途径。
如所讨论的,本公开内容涉及通过给予受试者广泛水解酪蛋白和/或其级分来调节特定激酶活性的方法。更具体地,在某些实施方案中,eHC包括包含SEQ ID NO 4, SEQ IDNO 13, SEQ ID NO 17, SEQ ID NO 21, SEQ ID NO 24, SEQ ID NO 30, SEQ ID NO 31,SEQ ID NO 32, SEQ ID NO 51, SEQ ID NO 57, SEQ ID NO 60和SEQ ID NO 63的肽组分。在一些实施方案中,所述肽组分可以包含表1中公开的另外的肽。例如,所述组合物可以包括表1中公开的至少10种另外的肽。
在另一个实施方案中,所述eHC还包括肽组分,所述肽组分包含至少3种选自SEQID NO 4, SEQ ID NO 13, SEQ ID NO 17, SEQ ID NO 21, SEQ ID NO 24, SEQ ID NO30, SEQ ID NO 31, SEQ ID NO 32, SEQ ID NO 51, SEQ ID NO 57, SEQ ID NO 60和SEQID NO 63的肽,以及至少5种选自表1的另外的肽。
下表1鉴定了可以包括在本公开内容的eHC中的特定氨基酸序列。
表1
下表2进一步鉴定了可包括在本文公开的eHC中和/或构成本文公开的eHC的来自表1的氨基酸序列子集。
表2
营养组合物
如提到的,eHC可被掺入或添加到营养组合物,特别是婴儿配方中。将营养组合物给予(例如,喂食)至受试者构成调节受试者中的激酶的方法。
因此,本公开内容一般涉及包含蛋白质等同物源的营养组合物,其中至少1%的蛋白质等同物源包含eHC,并且至多99%的蛋白质等同物源包含完整蛋白、部分水解蛋白、氨基酸或其组合。在实施方案中,1%至80%的蛋白质等同物源包含eHC,并且20%至99%的蛋白质等同物源包含完整蛋白、部分水解蛋白、氨基酸或其组合。在其它实施方案中,40%至100%的蛋白质等同物源包含eHC,0至60%的蛋白质等同物源包含完整蛋白、部分水解蛋白、氨基酸或其组合。在其它实施方案中,40%至70%的蛋白质等同物源包含eHC,并且30%至60%的蛋白质等同物源包含完整蛋白、部分水解蛋白、氨基酸或其组合。
在另一个实施方案中,20%至80%的蛋白质等同物源包括肽组分,其包含至少3种选自SEQ ID NO 4, SEQ ID NO 13, SEQ ID NO 17, SEQ ID NO 21, SEQ ID NO 24, SEQID NO 30, SEQ ID NO 31, SEQ ID NO 32, SEQ ID NO 51, SEQ ID NO 57, SEQ ID NO60和SEQ ID NO 63的肽,和至少5种选自表1的另外的肽。
在一些实施方案中,eHC可以以约0.2g/100kcal至约5.6g/100kcal的量存在于营养组合物中。在其它实施方案中,eHC可以以约1g/100kcal至约4g/100kcal的量存在于营养组合物中。在其它实施方案中,eHC可以以约2g/100kcal至约3g/100kcal的量存在于营养组合物中。
本文公开的蛋白质等同物源可以与营养组合物中的其它成分一起配制,以为目标受试者提供适当的营养水平。在一些实施方案中,蛋白质等同物源被包含在适合于支持正常生长的营养完全配方中。
在其它实施方案中,营养组合物可包含可添加至其它营养制剂的营养补剂或添加剂,所述营养制剂包括但不限于,食品和/或饮料。出于本公开内容的目的,“营养补剂”包括浓缩的营养物源,例如本文中鉴定的肽,或者其目的是补充正常饮食的具有营养或生理效果的其它物质。
如所讨论的,可以提供eHC作为蛋白质等同物源的要素。在一些实施方案中,表1和2中鉴定的肽可以通过水解获得,或者它们可以通过本领域技术人员已知的方法在体外合成。在Rangavajla等人的美国专利号7,618,669中公开了使用蛋白水解酶的水解方法的非限制性实例,该专利通过引用整体并入本文。然而,在本公开内容的实践中可以使用其它水解方法。
在一些实施方案中,蛋白质等同物源包含水解蛋白,例如酪蛋白,其包括部分水解蛋白和广泛水解蛋白(即,eHC)。在一些实施方案中,eHC包含广泛水解酪蛋白和/或其级分,包括摩尔质量分布大于500道尔顿的肽。在一些实施方案中,eHC包含具有约500道尔顿至约1,500道尔顿的范围的摩尔质量分布的肽。而且,在一些实施方案中,eHC可以包含具有约500道尔顿至约2,000道尔顿的摩尔质量分布范围的肽。
在一些实施方案中,蛋白质等同物源包含具有小于40%的水解度的部分水解蛋白。在其它实施方案中,蛋白质等同物源可以包含具有小于25%或小于15%的水解度的部分水解蛋白。
在一个具体的实施方案中,除了eHC以外,营养组合物不含蛋白质且含有游离氨基酸作为蛋白质等同物源。在该实施方案中,氨基酸可以包括但不限于组氨酸、异亮氨酸、亮氨酸、赖氨酸、甲硫氨酸、半胱氨酸、苯丙氨酸、酪氨酸、苏氨酸、色氨酸、缬氨酸、丙氨酸、精氨酸、天冬酰胺、天冬氨酸、谷氨酸、谷氨酰胺、甘氨酸、脯氨酸、丝氨酸、肉毒碱、牛磺酸及其混合物。在一些实施方案中,氨基酸可以是支链氨基酸。在其它实施方案中,可以包含小氨基酸肽作为营养组合物的蛋白质组分。这样的小氨基酸肽可以是天然存在的或合成的。营养组合物中游离氨基酸的量可以在约1至约5g/100kcal之间变化。在一个实施方案中,100%的游离氨基酸具有小于500道尔顿的分子量。在该实施方案中,营养组合物可以是低变应原性的。
在一个实施方案中,当蛋白质等同物源包含完整蛋白时,完整蛋白包含约40%至约85%乳清蛋白和约15%至约60%酪蛋白。
在一些实施方案中,营养组合物每100kcal包含约1g至约7g蛋白质等同物源。在其它实施方案中,营养组合物每100kcal包含约3.5g至约4.5g的蛋白质等同物源。
包含eHC的本公开内容的营养组合物可以每日一个或多个剂量给予。本公开内容预期任何口服可接受的剂型。这种剂型的实例包括但不限于丸剂、片剂、胶囊剂、软凝胶剂、液体剂、液体浓缩剂、粉剂、酏剂、溶液剂、混悬剂、乳剂、锭剂、珠剂、扁囊剂及其组合。
在一些实施方案中,可以将包含eHC的蛋白质等同物源添加到更完全的营养产品中。在该实施方案中,营养组合物可以含有脂肪或脂质和碳水化合物来源或组分,并且可以用于补充饮食或可以用作唯一的营养来源。
在一些实施方案中,营养组合物包含至少一种碳水化合物。碳水化合物可以是本领域使用的任一种,例如乳糖、葡萄糖、果糖、玉米糖浆固体、麦芽糖糊精、蔗糖、淀粉、稻米糖浆固体等。营养组合物中碳水化合物组分的量通常可以在约5g/100kcal至约25g/100kcal之间变化。在一些实施方案中,碳水化合物的量在约6g/100kcal至约22g/100kcal之间。在其它实施方案中,碳水化合物的量在约12g/100kcal至约14g/100kcal之间。在一些实施方案中,玉米糖浆固体是优选的。而且,水解的、部分水解的和/或广泛水解的碳水化合物由于它们的易消化性,对于包含在营养组合物中可以是合乎需要的。特别地,水解碳水化合物不太可能含有致敏的抗原表位。
适用于本文的碳水化合物材料的非限制性实例包括来源于玉米、木薯、稻米或马铃薯的水解或完整的、天然或化学改性的淀粉(蜡质性或非-蜡质性形式)。合适的碳水化合物的非限制性实例包括表征为水解玉米淀粉、麦芽糊精、麦芽糖、玉米糖浆、右旋糖、玉米糖浆固体、葡萄糖和各种其它葡萄糖聚合物及其组合的各种水解淀粉。其它合适的碳水化合物的非限制性实例包括通常称为蔗糖、乳糖、果糖、高果糖玉米糖浆、不消化的寡糖(例如寡聚果糖)及其组合的那些。
在一个具体的实施方案中,营养组合物的碳水化合物组分包含100%乳糖。在另一个实施方案中,另外的碳水化合物组分包含介于约0%和60%之间的乳糖。在另一个实施方案中,碳水化合物组分包含介于约15%和55%之间的乳糖。在又另一个实施方案中,碳水化合物组分包含介于约20%和30%之间的乳糖。在这些实施方案中,剩余的碳水化合物来源可以是本领域已知的任何碳水化合物。在一个实施方案中,碳水化合物组分包含约25%乳糖和约75%玉米糖浆固体。
在一些实施方案中,碳水化合物可包含至少一种淀粉或淀粉组分。淀粉是由两个不同的聚合物部分组成的碳水化合物:直链淀粉和支链淀粉。直链淀粉是由α-1,4连接的葡萄糖单元组成的线性部分。支链淀粉具有与直链淀粉相同的结构,但一些葡萄糖单元以α-1,6键化合,产生分支结构。淀粉一般含有17-24%的直链淀粉和76-83%的支链淀粉。还发展出了特殊遗传种类的植物,其产生具有不寻常的直链淀粉与支链淀粉的比例的淀粉。一些植物产生不含直链淀粉的淀粉。这些突变体在胚乳和花粉中产生淀粉粒,所述淀粉粒用碘染成红色并含有几乎100%的支链淀粉。这样的产生支链淀粉的植物主要是蜡质性玉米、蜡质性高粱和蜡质性稻米淀粉。
在加热、剪切和酸性条件下的淀粉性能可以经化学改性而改变或改进。改性通常通过引入化学取代基实现。例如,在高温或高剪切下的粘性可通过与二-或多官能团试剂,例如磷酰氯交联而增加或稳定。
在一些情况下,本公开内容的营养组合物包含至少一种经胶质化和/或预胶质化的淀粉。如本领域已知,当聚合物分子在其长度的一部分上相互作用以形成俘获溶剂和/或溶质分子的网络时发生胶质化。此外,当果胶分子由于共溶质(cosolute)分子的竞争性水合作用而失去水合的一些水时形成凝胶。影响胶质化发生的因素包括pH、共溶质的浓度、阳离子的浓度和类型、温度和果胶浓度。特别地,LM果胶会仅仅在二价阳离子,例如钙离子的存在下成凝胶。此外在LM果胶中,具有最低酯化程度的那些具有最高的胶凝温度和对用于交叉桥接(crossbridging)的二价阳离子的最大需要。
与此同时,淀粉的预胶质化是预煮淀粉以产生在冷水中水合和溶胀的物质的过程。然后干燥预煮的淀粉,例如通过转鼓式干燥或喷雾干燥。此外,本公开内容的淀粉可经化学改性以进一步扩展其最终特性的范围。本公开内容的营养组合物可包含至少一种预胶质化淀粉。
天然淀粉颗粒是不溶于水的,但当在水中加热时,当存在足够的热能以克服淀粉分子的键合力时,天然淀粉颗粒开始溶胀。随着继续加热,颗粒溶胀至其原始体积的许多倍。这些溶胀的颗粒之间的摩擦力是对淀粉糊粘性作出贡献的主要因素。
本公开内容的营养组合物可包含天然或改性淀粉,例如,蜡质性玉米淀粉、蜡质性稻米淀粉、玉米淀粉、稻米淀粉、马铃薯淀粉、木薯淀粉、小麦淀粉或其任何混合物。一般地,普通玉米淀粉包含约25%的直链淀粉,而蜡质性玉米淀粉几乎完全由支链淀粉组成。同时,马铃薯淀粉一般包含约20%的直链淀粉,稻米淀粉包含约20:80的直链淀粉:支链淀粉比例,和蜡质性稻米淀粉仅包含约2%的直链淀粉。此外,木薯淀粉一般包含约15%-约18%的直链淀粉,而小麦淀粉具有约25%的直链淀粉含量。
在一些实施方案中,营养组合物包含胶质化和/或预胶质化蜡质性玉米淀粉。在其它实施方案中,营养组合物包含胶质化和/或预胶质化木薯淀粉。其它胶质化或预胶质化淀粉,例如稻米淀粉或马铃薯淀粉也可使用。
用于本公开内容的营养组合物中的合适的脂肪或脂质可以是本领域已知或使用的任何脂肪或脂质,包括但不限于动物来源,例如乳脂、黄油、黄油脂肪、蛋黄脂质;海洋来源例如鱼油、海洋油、单细胞油;蔬菜和植物油例如玉米油、低芥酸菜子油(canola oil)、葵花油、大豆油、棕榈油精、椰子油、高油酸葵花籽油、月见草油、菜籽油、橄榄油、亚麻仁(亚麻籽)油、棉籽油、高油酸红花油、棕榈硬脂、棕榈仁油、小麦胚芽油;中链甘油三酯油以及脂肪酸的乳液和酯;及其任何组合。
在一个实施方案中,脂质或脂肪的量不大于约7g/100Kcal;在一些实施方案中,脂质或脂肪以约2至约7g/100Kcal的水平存在。
在某些实施方案中营养组合物还可含有一种或多种益生元(也被称为益生元组分)。益生元发挥健康益处,其可包括,但不限于,一种或有限数目的有益的肠道菌的生长和/或活性的选择性刺激,摄入的益生菌微生物的生长和/或活性的刺激,肠道病原体的选择性减少,和对肠道短链脂肪酸概况的有利的影响。这样的益生元可以是天然-存在的、合成的或通过生物体和/或植物的基因操作开发的,无论这种新的来源是现在已知的还是以后开发的。可用于本公开内容的益生元可包括寡糖、多糖,和含有果糖、木糖、大豆、半乳糖、葡萄糖和甘露糖的其它益生元。
更具体地说,可用于本公开内容的益生元可包括聚葡萄糖(PDX)、聚葡萄糖粉、乳果糖、乳蔗糖(lactosucrose)、棉子糖、寡聚葡萄糖、菊糖、寡聚果糖(FOS)、寡聚异麦芽糖、大豆寡糖、乳蔗糖、寡聚木糖(XOS)、寡聚壳糖、寡聚甘露糖、寡聚阿拉伯糖、寡聚唾液酸糖、寡聚岩藻糖、寡聚半乳糖(GOS)和寡聚龙胆糖。
在一个实施方案中,营养组合物中存在的益生元的总量可以为约1.0g/L至约10.0g/L的组合物。更优选地,营养组合物中存在的益生元的总量可以为约2.0g/L至约8.0g/L的组合物。在一些实施方案中,营养组合物中存在的益生元的总量可以为约0.01g/100kcal至约1.5g/100kcal。在某些实施方案中,营养组合物中存在的益生元的总量可以为约0.15g/100kcal至约1.5g/100kcal。而且,营养组合物可以包含含有PDX的益生元组分。在一些实施方案中,益生元组分包含至少20%w/w的PDX、GOS或其混合物。
在一个实施方案中,营养组合物中的PDX的量可以在约0.015g/100kcal至约1.5g/100kcal的范围内。在另一个实施方案中,聚葡萄糖的量在约0.2g/100kcal至约0.6g/100kcal的范围内。在一些实施方案中,PDX可以足以提供约1.0g/L至10.0g/L的量包含在营养组合物中。在另一个实施方案中,营养组合物含有约2.0g/L至8.0g/L的量的PDX。而在其它实施方案中,营养组合物中PDX的量可以为约0.05g/100kcal至约1.5g/100kcal。
在一些实施方案中,益生元组分还包含GOS。在一个实施方案中,营养组合物中的GOS的量可以为约0.015g/100kcal至约1.0g/100kcal。在另一个实施方案中,营养组合物中GOS的量可以为约0.2g/100kcal至约0.5g/100kcal。
在本公开内容的具体实施方案中,将PDX与GOS组合给予。
在一个具体的实施方案中,GOS和PDX以至少约0.015g/100kcal或约0.015g/100kcal至约1.5mg/100kcal的总量补充到营养组合物中。在一些实施方案中,营养组合物可以包含总量为约0.1至约1.0mg/100kcal的GOS和PDX。
乳铁蛋白也包括在本公开内容的营养组合物的一些实施方案中。乳铁蛋白是约80kD的单链多肽,其依据种类含有1-4个聚糖。不同种类的乳铁蛋白的3-D结构是极其相似的,但并不相同。每个乳铁蛋白包含两个同源叶,分别称为N-和C-叶,指分子的N-末端和C-末端部分。每个叶进一步由两个亚-叶或结构域组成,形成裂缝,其中铁离子(Fe3+)以协同合作的方式与碳酸(氢)根阴离子紧紧地结合。这些结构域分别被称为N1、N2、C1和C2。乳铁蛋白的N-末端具有强阳离子肽区域,其负责许多重要的结合特性。乳铁蛋白具有非常高的等电点(~pI 9)并且它的阳离子性质在其防御细菌、病毒和真菌的病原体的能力中起着主要作用。在乳铁蛋白的N-末端区域内存在若干阳离子氨基酸残基簇,其介导乳铁蛋白抗广泛范围的微生物的生物活性。例如,人乳铁蛋白的N-末端残基1-47 (牛乳铁蛋白的1-48)对乳铁蛋白的铁-非依赖性生物活性是至关重要的。在人乳铁蛋白中,残基2-5 (RRRR)和28-31 (RKVR)是在对乳铁蛋白的抗微生物活性特别重要的N-末端中富含精氨酸的阳离子结构域。N-末端中的类似的区域存在于牛乳铁蛋白中(残基17-42;FKCRRWQWRMKKLGAPSITCVRRAFA)。
来自不同的宿主种类的乳铁蛋白在它们的氨基酸序列中可以变化,但是通常具有相对高的等电点,伴有在内部叶的末端区域的带正电荷的氨基酸。用于本公开内容的合适的非人乳铁蛋白包括,但不限于,与人乳铁蛋白的氨基酸序列具有至少48%同源性的那些。例如,牛乳铁蛋白(“bLF”)具有这样的氨基酸组成,其具有与人乳铁蛋白约70%序列同源性。在一些实施方案中,非人乳铁蛋白具有与人乳铁蛋白至少55%的同源性,而在一些实施方案中,为至少65%同源性。用于本公开内容的可接受的非人乳铁蛋白包括,但不限于,bLF、猪乳铁蛋白、马乳铁蛋白、水牛乳铁蛋白、山羊乳铁蛋白、鼠乳铁蛋白和骆驼乳铁蛋白。
在一个实施方案中,乳铁蛋白以至少约15mg/100kCal的量存在于营养组合物中。在某些实施方案中,营养组合物可以包含每100kCal约15至约300mg乳铁蛋白。在另一个实施方案中,当营养组合物是婴儿配方时,营养组合物可以包含乳铁蛋白,其量为每100kCal约60mg至约150mg乳铁蛋白;在又一个实施方案中,营养组合物可以包含每100kCal约60mg至约100mg乳铁蛋白。
在一些实施方案中,营养组合物每毫升配方可以包含约0.5mg至约1.5mg的量的乳铁蛋白。在替代人乳的营养组合物中,乳铁蛋白可以以约0.6mg至约1.3mg/毫升配方的量存在。在某些实施方案中,营养组合物每升可以包含约0.1至约2克乳铁蛋白。在一些实施方案中,营养组合物每升配方包含约0.6至约1.5克乳铁蛋白。
在某些实施方案中使用的bLF可以是从全乳中分离和/或具有低体细胞计数的任何bLF,其中“低体细胞计数”是指小于200,000个细胞/mL的体细胞计数。举例来说,合适的bLF可获自Tatua Co-operative Dairy Co. Ltd., Morrinsville, New Zealand;FrieslandCampina Domo, Amersfoort, Netherlands;或Fonterra Co-Operative GroupLimited, Auckland, New Zealand。
用于本公开内容的乳铁蛋白可以,例如,从非人动物的乳中分离或通过经遗传修饰的生物体生产。例如,在美国专利号4,791,193(其通过引用以其全文结合到本文中)中,Okonogi等人公开制备高纯度的牛乳铁蛋白的方法。一般来说,所公开的方法包括三个步骤。首先使原料乳材料与弱酸性阳离子交换剂接触以吸附乳铁蛋白,接着进行洗涤以除去未吸附的物质的第二个步骤。随后进行解吸步骤,其中乳铁蛋白被取出以产生纯化的牛乳铁蛋白。其它方法可包括如在美国专利号7,368,141、5,849,885、5,919,913和5,861,491中描述的步骤,所述专利的公开内容都通过引用以其全文结合到本文中。
在某些实施方案中,用于本公开内容的乳铁蛋白可通过用于从乳源分离蛋白的膨胀床吸附(“EBA”)方法提供。EBA,有时也称为稳定的流化床吸附,是从乳源分离乳蛋白,如乳铁蛋白的方法,该方法包括建立包含颗粒基质的膨胀床吸附柱,将乳源施加于所述基质,并用包含约0.3-约2.0 M氯化钠的洗脱缓冲液从基质洗脱乳铁蛋白。任何哺乳动物乳源可用于本方法,而在具体实施方案中,乳源是牛乳来源。在一些实施方案中,乳源包括全脂乳、减脂乳、脱脂乳、乳清、酪蛋白、或其混合物。
在具体实施方案中,目标蛋白是乳铁蛋白,而其它乳蛋白,如乳过氧化物酶或乳清蛋白,也可被分离。在一些实施方案中,该方法包括以下步骤:建立包含颗粒基质的膨胀床吸附柱,将乳源施加于基质,并用约0.3-约2.0 M氯化钠从基质洗脱乳铁蛋白。在其它实施方案中,乳铁蛋白用约0.5-约1.0 M氯化钠洗脱,而在另外的实施方案中,乳铁蛋白用约0.7-约0.9 M氯化钠洗脱。
膨胀床吸附柱可以是本领域任何已知的,例如描述于美国专利号7,812,138、6,620,326和6,977,046中的那些,所述专利的公开内容藉此通过引用结合到本文中。在一些实施方案中,将乳源以膨胀模式施加于柱,并以膨胀或填充模式进行洗脱。在具体实施方案中,洗脱以膨胀模式进行。例如,膨胀模式的膨胀比例可以是约1-约3,或约1.3-约1.7。EBA技术在国际申请公布号WO 92/00799、WO 02/18237、WO 97/17132中进一步描述,所述申请藉此通过引用以其全文结合到本文中。
乳铁蛋白的等电点是大约8.9。分离乳铁蛋白的现有技术EBA方法使用200 mM氢氧化钠作为洗脱缓冲液。因此,系统的pH升至12以上,而乳铁蛋白的结构和生物活性可因不可逆的结构改变而受损。现在已经发现氯化钠溶液可用作从EBA基质分离乳铁蛋白的洗脱缓冲液。在某些实施方案中,氯化钠具有约0.3 M-约2.0 M的浓度。在其它实施方案中,乳铁蛋白洗脱缓冲液具有约0.3 M-约1.5 M,或约0.5 m-约1.0 M的氯化钠浓度。
在某些实施方案中,本公开内容的营养组合物还可以含有LCPUFA的来源;特别是含有DHA的LCPUFA的来源。其它合适的LCPUFA包括但不限于,α亚油酸、γ亚油酸、亚油酸、亚麻酸、二十碳五烯酸(EPA)和ARA。实际上,DHA和/或ARA可以与肌醇协同作用以进一步改善神经系统的健康和发育。
在一个实施方案中,特别是如果营养组合物是婴儿配方时,营养组合物补充有DHA和ARA二者。在该实施方案中,ARA: DHA的重量比可以在约1: 3至约9: 1之间。在一个具体的实施方案中,ARA: DHA的比率为约1: 2至约4: 1。
长链多不饱和脂肪酸在营养组合物中的量有利地为至少约5mg/100kcal,和可从约5mg/100kcal至约100mg/100kcal,更优选约10mg/100kcal至约50mg/100kcal变化。
营养组合物可以使用本领域已知的标准技术补充含有DHA和/或ARA的油。例如,DHA和ARA可以通过替换等量的油,例如通常存在于组合物中的高油酸向日葵油而添加到组合物中。作为另一个实例,含有DHA和ARA的油可以通过替换不含DHA和ARA的组合物中通常存在的等量的其余总脂肪共混物而添加到组合物中。
如果使用的话,DHA和/或ARA的来源可以是本领域已知的任何来源,例如海洋油、鱼油、单细胞油、蛋黄脂质和脑脂质。在一些实施方案中,DHA和ARA来源于单细胞Martek油、DHASCO® 和ARASCO®或其变体。DHA和ARA可以是天然形式,只要LCPUFA来源的其余部分不会对婴儿产生任何实质性的有害作用。或者,DHA和ARA可以以精制的形式使用。
在一个实施方案中,DHA和ARA的来源是单细胞油,如美国专利号5,374,567;5,550,156和5,397,591教导的,其公开内容通过引用整体并入本文。然而,本公开内容不限于仅这些油。
在一些实施方案中,营养组合物可以包含源自乳的富集的脂质部分。来源于乳的富集的脂质部分可以通过任何数量的分级分离技术来生产。这些技术包括但不限于熔点分级分离、有机溶剂分级分离、超临界流体分级分离以及其任何变体和组合。在一些实施方案中,营养组合物可以包含源自含有乳脂肪球的乳的富集的脂质部分。
在某些实施方案中,添加富集的脂质部分或含有乳脂肪球的富集的脂质部分可以给营养组合物提供饱和脂肪酸、反式脂肪酸、单不饱和脂肪酸、多不饱和脂肪酸、奇数支链脂肪酸(OBCFAs)、支链脂肪酸(BCFAs)、(共轭亚油酸)CLA、胆固醇、磷脂和/或乳脂肪球膜(MFGM)以及MFGM蛋白的来源。
乳脂肪球可以具有至少约2μm的平均直径(体积-表面积平均直径)。在一些实施方案中,平均直径在约2μm至约13μm的范围内。在其它实施方案中,乳脂肪球可以在约2.5μm至约10μm的范围内。仍然在其它实施方案中,乳脂肪球的平均直径可以在约3μm至约6μm的范围内。在某些实施方案中,球的比表面积小于3.5m2/g,并且在其它实施方案中,在约0.9m2/g至约3m2/g之间。不受任何特定理论的约束,认为上述大小的乳脂肪球更易于接近脂肪酶,因此导致更好的脂质消化。
在一些实施方案中,富集的脂质部分和/或乳脂肪球含有饱和脂肪酸。饱和脂肪酸可以以约0.1g/100kcal至约8.0g/100kcal的浓度存在。在某些实施方案中,饱和脂肪酸可以以约0.5g/100kcal至约2.0g/100kcal的量存在。在其它实施方案中,饱和脂肪酸可以约3.5g/100kcal至约6.9g/100kcal存在。
适合包含的饱和脂肪酸的实例包括但不限于丁酸、戊酸、己酸、辛酸、癸酸、月桂酸、肉豆蔻酸、棕榈酸、硬脂酸、花生酸、山俞酸、木蜡酸、十四酸、十六酸、棕榈酸和十八酸、和/或其组合和混合物。
另外,在一些实施方案中,富集的脂质部分和/或乳脂肪球可包含月桂酸。月桂酸也被称为十二烷酸,是具有12个碳原子链的饱和脂肪酸,被认为是目前在人母乳中发现的主要抗病毒和抗菌物质之一。乳脂肪球可以富含在Sn-1、Sn-2和/或Sn-3位置含有月桂酸的甘油三酯。不受任何特定理论的束缚,认为当摄入富集的脂质部分时,口腔舌脂肪酶和胰脂肪酶将甘油三酯水解成包括单月桂酸的甘油酯和游离月桂酸的混合物。
月桂酸在球中的浓度从80mg/100ml至800mg/100ml变化。球中单月桂基的浓度可以在20mg/100ml至300mg/100ml进料的范围内。在一些实施方案中,范围是60mg/100ml至130mg/100ml。
在某些实施方案中,富集的脂质部分和/或乳脂肪球可以含有反式脂肪酸。包含在乳脂肪球中的反式脂肪酸可以是单不饱和或多不饱和反式脂肪酸。在一些实施方案中,反式脂肪酸可以以约0.2g/100kcal至约7.0g/100kcal的量存在。在其它实施方案中,反式脂肪酸可以以约3.4g/100kcal至约5.2g/100kcal的量存在。在其它实施方案中,反式脂肪酸可以以约1.2g/100kcal至约4.3g/100kcal的量存在。
用于包含的反式脂肪酸的实例包括但不限于,异油酸或反油酸及其混合物。此外,当哺乳动物消耗时,将异油酸转化为瘤胃酸,其是具有抗癌性质的共轭亚油酸。此外,富含异油酸的饮食可以帮助降低总胆固醇、LDL胆固醇和甘油三酯水平。
在一些实施方案中,富集的脂质部分和/或乳脂肪球可以包含OBCFA。在某些实施方案中,OBCFA可以以约0.3g/100kcal至约6.1g/100kcal的量存在。在其它实施方案中,OBCFA可以以约2.2g/100kcal至约4.3g/100kcal的量存在。在又一个实施方案中,OBCFA可以以约3.5g/100kcal至约5.7g/100kcal的量存在。在其它实施方案中,乳脂肪球包含至少一种OBCFA。
通常,婴儿可以在哺乳母亲的子宫内和从母乳中吸收OBCFA。因此,在人乳中鉴定的OBCFA优选包含在营养组合物的乳脂肪球中。将OBCFA添加到婴儿或儿童的配方允许这样的配方反映人乳的组成和功能,并促进一般健康和康乐。
在一些实施方案中,富集的脂质部分和/或乳脂肪球可以包含BCFA。在一些实施方案中,BCFA以约0.2g/100kcal至约5.82g/100kcal的浓度存在。在另一个实施方案中,BCFA以约2.3g/100kcal至约4.2g/100kcal的量存在。在又一个实施方案中,BCFA以约4.2g/100kcal至约5.82g/100kcal存在。在其它实施方案中,乳脂肪球包含至少一种BCFA。
在人乳中鉴定的BCFA优选包含在营养组合物中。将BCFA添加到婴儿或儿童配方允许这样的配方反映人乳的组成和功能,并促进一般健康和康乐。
在某些实施方案中,富集的脂质部分和/或乳脂肪球可包含CLA。在一些实施方案中,CLA可以以约0.4g/100kcal至约2.5g/100kcal的浓度存在。在其它实施方案中,CLA可以约0.8g/100kcal至约1.2g/100kcal存在。在其它实施方案中,CLA可以约1.2g/100kcal至约2.3g/100kcal存在。在其它实施方案中,乳脂肪球包含至少一种CLA。
在人乳中鉴定的CLA优选包含在营养组合物中。一般来说,CLA被婴儿从哺乳母亲的人乳吸收。将CLA添加到婴儿或儿童配方允许这样的配方反映人乳的组成和功能,并促进一般健康和康乐。
存在于营养组合物的乳脂肪球中的CLA的实例包括但不限于顺-9,反-11 CLA、反-10,顺-12 CLA、顺-9,反-12-十八碳二烯酸和其混合物。
在一些实施方案中,本公开内容的富集的脂质部分和/或乳脂肪球包含单不饱和脂肪酸。富集的脂质部分和/或乳脂肪球可以配制成包括约0.8g/100kcal至约2.5g/100kcal的单不饱和脂肪酸。在其它实施方案中,乳脂肪球可包含约1.2g/100kcal至约1.8g/100kcal的单不饱和脂肪酸。
单不饱和脂肪酸的实例包括但不限于棕榈油酸、顺式-异油酸、油酸及其混合物。
在某些实施方案中,本公开内容的富集的脂质部分和/或乳脂肪球包含约2.3g/100kcal至约4.4g/100kcal的多不饱和脂肪酸。在其它实施方案中,多不饱和脂肪酸以约2.7g/100kcal至约3.5g/100kcal存在。在又一个实施方案中,多不饱和脂肪酸以约2.4g/100kcal至约3.3g/100kcal存在。
在一些实施方案中,本公开内容的富集的脂质部分和/或乳脂肪球包含多不饱和脂肪酸、例如亚油酸、亚麻酸、十八碳三烯酸、花生四烯酸(ARA)、二十碳四烯酸、二十碳五烯酸(EPA)、二十二碳五烯酸(DPA)和二十二碳六烯酸(DHA)。多不饱和脂肪酸是前列腺素和类二十烷酸的前体,已知其提供许多健康益处,包括抗炎反应、胆固醇吸收和增加的支气管功能。
在一些实施方案中,本公开内容的富集的脂质部分和/或乳脂肪球还可以包含胆固醇,水平为约100mg/100kcal至约400mg/100kcal。在另一个实施方案中,胆固醇以约200mg/100kcal至约300mg/100kcal存在。与人乳和牛乳相似,包含在乳脂肪球中的胆固醇可以存在于乳脂肪球的外双层膜中以提供对球状膜的稳定性。
在一些实施方案中,本公开内容的富集的脂质部分和/或乳脂肪球包含约50mg/100kcal至约200mg/100kcal磷脂。在其它实施方案中,磷脂以约75mg/100kcal至约150mg/100kcal存在。在其它实施方案中,磷脂以约100mg/100kcal至约250mg/100kcal的浓度存在。
在某些实施方案中,可通过提供磷脂膜或双层磷脂膜将磷脂掺入乳脂肪球中以稳定乳脂球。因此,在一些实施方案中,可以用比人乳中发现的更高量的磷脂配制乳脂肪球。
已经测量了人乳脂质的磷脂组成(以总磷脂的重量百分比计)为磷脂酰胆碱(“PC”) 24.9%, 磷脂酰乙醇胺(“PE”) 27.7%, 磷脂酰丝氨酸(“PS”) 9.3%, 磷脂酰肌醇(“PI”) 5.4%和鞘磷脂(“SM”) 32.4%, (Harzer, G. et al., canAm. J. Clin. Nutr.,Vol. 37, pp. 612-621 (1983))。因此在一个实施方案中,乳脂肪球包含PC,PE,PS,PI,SM中的一种或多种及其混合物。此外,包含在乳脂肪球中的磷脂组合物可以通过掺入所需的磷脂来配制以提供某些健康益处。
在某些实施方案中,本公开内容的富集的脂质部分和/或乳脂肪球包含乳脂肪球膜蛋白。在一些实施方案中,乳脂肪球膜蛋白以约50mg/100kcal至约500mg/100kcal存在。
在一些实施方案中,在本公开内容的富集的脂质部分和/或乳脂肪球中可以包括半乳糖脂。为了本公开内容的目的,“半乳糖脂”是指其糖基团是半乳糖的任何糖脂。更具体地说,半乳糖脂与糖鞘脂不同之处在于它们在其组成中不含氮。半乳糖脂在支持大脑发育和整体神经元健康方面发挥重要作用。此外,半乳糖脂、半乳糖脑苷脂和硫苷脂分别占髓磷脂总含量的约23%和4%,因此在一些实施方案中,可以掺入乳脂肪球中。
此外,本公开内容的营养组合物包含至少一种果胶来源。果胶来源可包含本领域已知的任何种类或等级的果胶。在一些实施方案中,果胶具有少于50%的酯化程度并被分类为低甲基化(“LM”)果胶。在一些实施方案中,果胶具有大于或等于50%的酯化程度和被分类为高-酯或高甲基化(“HM”)果胶。在还有的其它实施方案中,果胶为极低(“VL”)果胶,其具有少于约15%的酯化程度。此外,本公开内容的营养组合物可包含LM果胶、HM果胶、VL果胶或其任何混合物。营养组合物可包括可溶于水的果胶。此外如本领域已知的,果胶溶液的溶解性和粘性与分子量、酯化程度、果胶制剂的浓度以及相反离子的pH和存在有关。
而且,果胶具有形成凝胶的独特能力。一般来说,在类似的条件下,果胶的胶凝程度、胶凝温度和凝胶强度彼此成比例,且各自一般与果胶的分子量成比例和与酯化程度成反比。例如,当果胶溶液的pH降低时,羧酸基团的离子化受到抑制,结果是失去其电荷,糖分子在其整个长度上不互相排斥。因此,多糖分子可在其长度的一部分上缔合以形成凝胶。但具有增加的甲基化程度的果胶会在稍微更高的pH下胶凝,因为它们在任何给定pH下具有较少的羧酸根阴离子(J.N. Bemiller,对果胶的介绍:结构和特性(Introduction to Pectins: Structure and Properties), 果胶的化学和功能(Chemistry and Functionof Pectins);第1章;1986)。
营养组合物可包含与果胶和/或胶质化果胶一起的胶质化淀粉和/或预胶质化淀粉。虽然不希望受到任何理论的束缚,相信与淀粉颗粒一起使用果胶,例如LM果胶(其是大分子量的水状胶体),提供增加流体基质中分子内摩擦力的协同作用。果胶的羧基也可与存在于营养组合物中的钙离子相互作用,因而导致粘性的增加,因为果胶的羧基与钙离子,并且还与存在于营养组合物中的肽形成弱的凝胶结构。在一些实施方案中,营养组合物包含分别介于约12:1和20:1之间的比例的淀粉:果胶。在其它实施方案中,所述淀粉与果胶的比例是约17:1。在一些实施方案中,营养组合物可包含约0.05和约2.0% w/w之间的果胶。在一特定的实施方案中,营养组合物可包含约0.5% w/w的果胶。
本文使用的果胶通常具有8,000道尔顿或更大的峰分子量。本公开内容的果胶具有优选的介于8,000和约500,000之间,更优选介于约10,000和约200,000之间和最优选介于约15,000和约100,000道尔顿之间的峰分子量。在一些实施方案中,本公开内容的果胶可以是水解果胶,其分子量小于完整或未改性果胶的分子量。本公开内容的水解果胶可通过本领域已知的减少分子量的任何方法制备。实例包括化学水解、酶促水解和机械剪切。减少分子量的优选的方法是通过在升高的温度下进行碱性或中性水解。在一些实施方案中,营养组合物包含部分水解果胶。在某些实施方案中,部分水解果胶具有少于完整或未改性果胶的分子量但超过3,300道尔顿的分子量。
营养组合物可含有至少一种酸性多糖。酸性多糖,例如带负电荷的果胶,可诱导对受试者的胃肠道中的病原体的抗-粘附作用。事实上,衍生自果胶的非人乳酸性寡糖能够与上皮表面相互作用且已知抑制病原体在上皮表面上的附着。
在一些实施方案中,营养组合物包含至少一种果胶-衍生的酸性寡糖。果胶-衍生的酸性寡糖(pAOS)由酶促果胶分解(pectinolysis)产生,和pAOS的大小取决于所用的酶和反应的持续时间。在这样的实施方案中,pAOS可有益地影响受试者的粪便粘性、排便频率、粪便pH和/或喂养耐受性。本公开内容的营养组合物可包含约1 g pAOS每升的营养组合物-约6 g pAOS每升的营养组合物。
在一些实施方案中,营养组合物包含至多约20% w/w的淀粉和果胶的混合物。在一些实施方案中,营养组合物包含至多约19%的淀粉和至多约1%的果胶。在其它实施方案中,营养组合物包含至多约15%的淀粉和至多约5%的果胶。在还有的其它实施方案中,营养组合物包含至多约18%的淀粉和至多约2%的果胶。在一些实施方案中,营养组合物包含约0.05%w/w和约20% w/w之间的淀粉和果胶的混合物。其它实施方案包含约0.05%和约19% w/w之间的淀粉和约0.05%和约1% w/w之间的果胶。此外,营养组合物可包含约0.05%和约15% w/w之间的淀粉和约0.05%和约5% w/w之间的果胶。
在一些实施方案中,营养组合物包含唾液酸。唾液酸是超过50个9碳糖的成员的家族,所有这些都是神经氨酸的衍生物。在人类中发现的主要唾液酸家族来自N-乙酰神经氨酸亚家族。唾液酸存在于乳中,例如牛和山羊。在哺乳动物中,与其它体细胞膜相比,神经元细胞膜具有最高浓度的唾液酸。唾液酸残基也是神经节苷脂的组分。
如果包含在营养组合物中,则唾液酸可以以约0.5mg/100kcal至约45mg/100kcal的量存在。在一些实施方案中,唾液酸可以以约5mg/100kcal至约30mg/100kcal的量存在。在其它实施方案中,唾液酸可以以约10mg/100kcal至约25mg/100kcal的量存在。
在一个实施方案中,营养组合物可含有一种或多种益生菌。本领域已知的任何益生菌可以是这个实施方案中可接受的。在具体的实施方案中,益生菌可以选自任何乳酸杆菌菌种(Lactobacillus species)例如鼠李糖乳酸杆菌GG (LGG) (ATCC号53103)、双歧杆菌菌种(Bifidobacterium species)例如长双歧杆菌(Bifidobacterium longum) BB536(BL999, ATCC: BAA-999)、长双歧杆菌(Bifidobacterium longum) AH1206 (NCIMB:41382)、短双歧杆菌(Bifidobacterium breve) AH1205 (NCIMB: 41387)、婴儿双岐杆菌(Bifidobacterium infantis) 35624 (NCIMB: 41003)和动物双歧杆菌产乳酸亚种BB-12(Bifidobacterium animalis subsp. lactis BB-12) (DSM No. 10140)或其任何组合。
如果包括在组合物中,益生菌的量可以从每100 Kcal约1 x 104至约1.5 x 1012cfu的益生菌变化。在一些实施方案中,益生菌的量可以为每100 Kcal约1 x 106至约1 x109 cfu的益生菌。在某些其它实施方案中,益生菌的量可从约1 x 107 cfu/100 Kcal至约1x 108 cfu每100 Kcal的益生菌变化。
在一个实施方案中,益生菌可以是有活力的或无活力的。如本文所用的,术语“有活力的”是指活的微生物体。术语“无活力的”或“无活力的益生菌”意指无生命的益生菌微生物、其细胞组成和/或其代谢物。这样的无活力的益生菌可以被热杀死或以别的方式灭活,但它们保留有利地影响宿主健康的能力。可用于本公开内容的益生菌可以是天然-存在的、合成的或通过生物体的基因操作开发的,无论这样的来源是现在已知的还是以后开发的。
在一些实施方案中,营养组合物可包括包含益生菌细胞等同物的来源,其指等同于相等数目的活细胞的无活力、不复制的益生菌的水平。术语“不复制”要理解为从相同量的复制细菌(cfu/g)获得的不复制微生物的量,包括灭活的益生菌、DNA片段、细胞壁或胞质化合物。换言之,非-存活的、不复制生物体的量以cfu来表示,就如所有的微生物都是活的,而不管它们是否是死的、不复制、灭活的、破碎的等。在无活力的益生菌包含在营养组合物中时,益生菌细胞等同物的量可从约1 x 104至约1.5 x 1010益生菌细胞等同物/100 Kcal变化。在一些实施方案中,益生菌细胞等同物的量可以是从约1 x 106至约1 x 109益生菌细胞等同物/100 Kcal营养组合物。在某些其它实施方案中,益生菌细胞等同物的量可以从约1 x 107-约1 x 108益生菌细胞等同物/100 Kcal营养组合物变化。
在一些实施方案中,掺入营养组合物的益生菌来源可包含有活力的菌落形成单元和无活力的细胞-等同物二者。
在一些实施方案中,营养组合物包含来自益生菌分批培养过程的指数生长后期的培养物上清液。不希望受到理论的束缚,相信培养物上清液的活性可归因于如发现的在益生菌分批培养的指数(或“对数”)期的后阶段释放到培养基中的组分(包括蛋白质材料,和可能包括(表)多糖材料)的混合物。本文所用的术语“培养物上清液”包括在培养基中存在的组分的混合物。在细菌的分批培养中公认的阶段是技术人员已知的。这些是“滞后”、“对数” (“对数的”或“指数的”)、“静止”和“死亡” (或“对数下降”)期。在活菌存在的所有阶段中,细菌从培养基中代谢营养物,并将材料分泌(施加、释放)到培养基中。在生长阶段的给定的时间点分泌的材料的组成一般是不可预测的。
在一个实施方案中,培养物上清液可通过包括以下步骤的方法获得:(a) 使用分批方法,使益生菌例如LGG在合适的培养基中培养;(b) 收获在培养步骤的指数生长后期的培养物上清液,该期参照分批-培养过程的滞后期和静止期之间的后半段时间限定;(c) 任选地从上清液除去低分子量成分以保留5-6千道尔顿(kDa)以上分子量的成分;(d) 从培养物上清液除去液体内容物以获得组合物。
培养物上清液可包含从指数期晚期收获的分泌材料。指数期晚期发生在中期指数期(其为指数期持续时间的中段时间,因此提及指数期晚期为滞后期和静止期之间的后半段时间)之后的时间。特别是,本文使用术语“指数期晚期”是指LGG分批-培养过程的滞后期和静止期之间时间的后四分之一部分。在一些实施方案中,培养物上清液在指数期持续时间的75%-85%的时间点收获,且可以在指数期中流逝时间的约5/6时收获。
如提到的,所公开的营养组合物可包含β-葡聚糖源。葡聚糖是多糖,具体而言是葡萄糖的聚合物,其是天然存在的并且可存在于细菌、酵母、真菌和植物的细胞壁中。β葡聚糖(β-葡聚糖)本身是不同子集的葡萄糖聚合物,其由通过β-型糖苷键连接在一起以形成复杂的碳水化合物的葡萄糖单体链构成。
β-1,3-葡聚糖是纯化自例如酵母、蕈类、细菌、藻类或谷类的碳水化合物聚合物。β-1,3-葡聚糖的化学结构取决于β-1,3-葡聚糖的来源。而且,各种物理化学参数,例如溶解度、主要结构、分子量和分支,在β-1,3-葡聚糖的生物活性中起作用。
β-1,3-葡聚糖是天然存在的多糖,含或不含β-1,6-葡萄糖侧链,其存在于各种植物、酵母、真菌和细菌的细胞壁中。β-1,3;1,6-葡聚糖是含有具有在(1,6)位连接的侧链并带有(1,3)键的葡萄糖单元的葡聚糖。β-1,3;1,6葡聚糖是共享结构共性的一组异源葡萄糖聚合物,包括通过β-1,3键连接的直链葡萄糖单元骨架与从该骨架延伸的β-1,6-连接的葡萄糖分支。虽然这是本发明所述β-葡聚糖类别的基本结构,但可能存在一些变化。例如,某些酵母β-葡聚糖具有从β(1,6)分支延伸出的额外的β(1,3)分支区域,这进一步增加了其相应结构的复杂性。
来源于面包酵母,酿酒酵母(Saccharomyces cerevisiae)的β-葡聚糖由在1和3位连接的D-葡萄糖分子链组成,其具有在1和6位上连接的葡萄糖侧链。酵母来源的β-葡聚糖是不溶性纤维样复合糖,具有如下普遍结构:带有β-1,3骨架的葡萄糖单元直链,所述骨架中间散布着长度一般为6-8个葡萄糖单元的β-1,6侧链。更具体地说,来源于面包酵母的β-葡聚糖是聚-(1,6)-β-D-吡喃葡糖基-(1,3)-β-D-吡喃葡萄糖。
此外,β-葡聚糖在儿科受试者中耐受性良好,不产生或引起过量产气、腹胀、胃气胀或腹泻。将β-葡聚糖添加至用于儿科受试者的营养组合物中,例如婴儿配方、成长乳或另一个儿童营养制品,会通过提高针对侵入的病原体的抗性来改善受试者的免疫应答,并因此维持或改善整体健康。
在一些实施方案中,存在于组合物中的β-葡聚糖的量在每100g组合物约0.010和约0.080 g之间。在其它实施方案中,营养组合物包含每份饮食约10和约30 mg之间的β-葡聚糖。在另一个实施方案中,营养组合物包含每8液量盎司(236.6 mL)份饮食约5和约30 mg之间的β-葡聚糖。在其它实施方案中,营养组合物包含足以提供每天约15 mg和约90 mg之间的β-葡聚糖的量的β-葡聚糖。营养组合物可以多个剂量递送,以达到一整天递送给受试者的目标量的β-葡聚糖。
在一些实施方案中,营养组合物中的β-葡聚糖的量在每100 Kcal约3 mg和约17mg之间。在另一个实施方案中,β-葡聚糖的量在每100 Kcal约6 mg和约17 mg之间。
还可以足以供应受试者每日营养需要的量将一种或多种维生素和/或矿物质加入营养组合物中。本领域普通技术人员将了解,维生素和矿物质需要会根据例如儿童的年龄而变化。例如,婴儿可具有与年龄介于1岁和13岁之间的儿童不同的维生素和矿物质需要。因此,实施方案无意将营养组合物限于具体的年龄组,而是提供一定范围的可接受的维生素和矿物质组分。
营养组合物可任选地包括但不限于下列维生素或其衍生物的一种或多种:维生素B1 (硫胺、焦磷酸硫胺,TPP、三磷酸硫胺,TTP、盐酸硫胺、一硝酸硫胺)、维生素B2 (核黄素、黄素单核苷酸,FMN、黄素腺嘌呤二核苷酸,FAD、乳黄素、卵黄素)、维生素B3 (尼克酸、烟酸、烟酰胺、尼克酰胺、烟酰胺腺嘌呤二核苷酸,NAD、烟酸单核苷酸,NicMN、吡啶-3-甲酸)、维生素B3-前体色氨酸、维生素B6 (吡哆醇、吡哆醛、吡哆胺、盐酸吡哆醇)、泛酸(泛酸盐、泛醇)、叶酸盐(叶酸(folic acid)、叶酸(folacin)、蝶酰谷氨酸)、维生素B12(钴胺素、甲钴胺、脱氧腺苷钴胺素、氰钴胺素、羟钴胺素、腺苷钴胺素)、生物素、维生素C (抗坏血酸)、维生素A(视黄醇、视黄醇乙酸酯、视黄醇棕榈酸酯、含有其它长链脂肪酸的视黄酯、视黄醛、视黄酸、视黄醇酯)、维生素D (钙化醇、胆钙化醇、维生素D3、1,25,-二羟维生素D)、维生素E (α-生育酚、α-生育酚乙酸酯、α-生育酚琥珀酸酯、α-生育酚烟酸酯、α-生育酚)、维生素K (维生素K1、叶绿醌、萘醌、维生素K2、甲基萘醌-7、维生素K3、甲基萘醌-4、甲萘醌、甲基萘醌-8、甲基萘醌-8H、甲基萘醌-9、甲基萘醌-9H、甲基萘醌-10、甲基萘醌-11、甲基萘醌-12、甲基萘醌-13)、胆碱、肌醇、β-胡萝卜素及其任何组合。
此外,营养组合物可以任选地包括但不限于,下列矿物质或其衍生物的一种或几种:硼、钙、醋酸钙、葡萄糖酸钙、氯化钙、乳酸钙、磷酸钙、硫酸钙、氯化物、铬、氯化铬、吡啶甲酸铬(chromium picolinate)、铜、硫酸铜(copper sulfate)、葡萄糖酸铜、硫酸铜(cupric sulfate)、氟化物、铁、羰基铁、三价铁、富马酸亚铁、正磷酸铁、铁研制剂(irontrituration)、多糖铁、碘化物、碘、镁、碳酸镁、氢氧化镁、氧化镁、硬脂酸镁、硫酸镁、锰、钼、磷、钾、磷酸钾、碘化钾、氯化钾、醋酸钾、硒、硫、钠、多库酯钠、氯化钠、硒酸钠、钼酸钠、锌、氧化锌、硫酸锌及其混合物。矿物质化合物的非限制性示例性衍生物包括任何矿物质化合物的盐、碱性盐、酯和螯合物。
矿物质可以以盐例如磷酸钙、甘油磷酸钙、柠檬酸钠、氯化钾、磷酸钾、磷酸镁、硫酸亚铁、硫酸锌、硫酸铜、硫酸镁和亚硒酸钠的形式添加至营养组合物中。可添加本领域已知的额外的维生素和矿物质。
在一个实施方案中,营养组合物每份可包含任何给定的国家的最大饮食推荐量的约10和约50%之间或一组国家的平均饮食推荐量的约10和约50%之间的维生素A、C和E、锌、铁、碘、硒和胆碱。在另一个实施方案中,儿童营养组合物每份可供给任何给定的国家的最大饮食推荐量的约10-30%或一组国家的平均饮食推荐量的约10-30%的维生素B族。在又另一个实施方案中,儿童营养产品中的维生素D、钙、镁、磷和钾的水平可与乳中存在的平均水平一致。在其它实施方案中,儿童营养组合物中的其它营养物每份可以以任何给定国家的最大饮食推荐量的约20%或一组国家的平均饮食推荐量的约20%存在。
本公开内容的营养组合物可以任选地包括下列调味剂中的一种或多种,其包括但不限于:调味提取物、挥发油类、可可或巧克力调味剂、花生酱调味剂、饼干屑、香草或任何可市售获得的调味剂。有用的调味剂的实例包括,但不限于,纯茴香提取物、仿香蕉提取物、仿樱桃提取物、巧克力提取物、纯柠檬提取物、纯橙子提取物、纯薄荷提取物、蜂蜜、仿菠萝提取物、仿朗姆酒(imitation rum)提取物、仿草莓提取物或香草提取物;或挥发油类,例如蜂蜜花油、月桂油、香柠檬油、柏木油、樱桃油、肉桂油、丁香油或薄荷油;花生酱、巧克力调味剂、香草饼干屑、奶油硬糖、太妃糖,及其混合物。调味剂的量可变化很大,这取决于使用的调味剂。可依照本领域已知的,选择调味剂的类型和量。
本公开内容的营养组合物可以任选地包括一种或多种可添加用于稳定最终产品的乳化剂。合适的乳化剂的实例包括但不限于,卵磷脂(例如来自蛋或大豆)、α乳清蛋白和/或甘油单酯和甘油二酯及其混合物。其它乳化剂对于技术人员是显而易见的,合适乳化剂的选择将部分取决于配方和终产品。
本公开内容的营养组合物可以任选地包含一种或多种也可添加以延长产品货架期的防腐剂。合适的防腐剂包括但不限于,山梨酸钾、山梨酸钠、苯甲酸钾、苯甲酸钠、EDTA钙二钠,及其混合物。
本公开内容的营养组合物可以任选地包含一种或多种稳定剂。用于实施本公开内容的营养组合物的合适的稳定剂包括但不限于,阿拉伯树胶、茄替胶、刺梧桐胶、西黄蓍胶、琼脂、帚叉藻聚糖、瓜尔豆胶、吉兰胶、槐豆胶、果胶、低甲氧基果胶、明胶、微晶纤维素、CMC(羧甲基纤维素钠)、甲基纤维素羟丙基甲基纤维素、羟丙基纤维素、DATEM (甘油单酯和甘油二酯的二乙酰酒石酸酯)、葡聚糖、角叉菜胶,及其混合物。
所公开的营养组合物可以本领域已知的任何形式提供,例如粉剂、凝胶剂、混悬剂、糊剂、固体剂、液体剂、液体浓缩物、可复溶奶粉代用品或即用制品。在某些实施方案中,营养组合物可包含营养补剂、儿童营养制品、婴儿配方、人乳强化剂、成长乳或设计用于婴儿或儿科受试者的任何其它的营养组合物。本公开内容的营养组合物包括例如可口服摄入的促进健康的物质,包括例如食品、饮料、片剂、胶囊剂和粉剂。而且,可使本公开内容的营养组合物标准化至特定的含热量,其可作为即用制品提供,或它可以浓缩形式提供。在一些实施方案中,营养组合物可呈粉末形式,其粒径范围为5 μm-1500 μm,更优选范围为10 μm-300 μm。
如果营养组合物呈即用制品的形式,营养组合物的重量克分子渗透浓度可介于约100和约1100 mOsm/kg水之间,更通常约200-约700 mOsm/kg水。
本公开内容的营养组合物还可包含至少一种另外的植物营养素,即除了上述果胶和/或淀粉组分之外另一种植物营养素组分。在人乳中鉴定的植物营养素,或其衍生物、共轭形式或前体,优选地包含在营养组合物中。典型地,类胡萝卜素和多酚的膳食来源被养育的母亲所吸收并保留在乳汁中,使得它们可为养育婴儿利用。加入这些植物营养素至婴儿或儿童的配方,以使这样的配方反映人乳的组成和功能并促进总体健康和良好状态。
例如,在一些实施方案中,本公开内容的营养组合物可在每份膳食8液量盎司(236.6 mL)中包含介于约80和约300 mg之间的花青苷,介于约100和约600 mg之间的原花青素,介于约50和约500 mg之间的黄烷-3-醇,或其任何组合或混合物。在其它实施方案中,营养组合物包含苹果提取物、葡萄籽提取物或其组合或混合物。而且,营养组合物的至少一种植物营养素可衍生自任何单一或共混的果实、葡萄籽和/或苹果或茶提取物。
为了本公开内容的目的,可将天然、纯化的、包囊的和/或化学或酶促-改性形式的额外植物营养素加入到营养组合物中,以递送所需感官和稳定性特性。在包囊的情况下,合乎需要的是,封囊的植物营养素抵抗水的溶解但在抵达小肠时释放。这可通过应用肠溶衣实现,所述肠溶衣例如交联藻酸盐等。
适合于营养组合物的另外的植物营养素的实例包括,但不限于,花青苷、原花青素、黄烷-3-醇(即儿茶素、表儿茶素等)、黄烷酮、类黄酮、异类黄酮、茋类(stilbenoids)(即白藜芦醇等)、原花青素、花青苷、白藜芦醇、槲皮素、姜黄素和/或其任何混合物,以及纯化的或天然形式的植物营养素的任何可能的组合。营养组合物的某些组分,特别是植物-基组分可提供植物营养素的来源。
某些量的植物营养素可以固有地存在于已知成分中,例如天然油,其通常用来制备用于儿科受试者的营养组合物。这些固有的植物营养素可以是但不必认为是本公开内容描述的植物营养素组分的一部分。在一些实施方案中,如本文描述的植物营养素浓度和比率基于加入的和固有的植物营养素源计算。在其它实施方案中,如本文描述的植物营养素浓度和比率仅基于加入的植物营养素源计算。
在一些实施方案中,营养组合物包含花青苷,例如,橙皮(aurantinidin)、矢车菊色素、飞燕草色素、欧天芥菜色素(europinidin)、木樨黄定(luteolinidin)、天竺葵色素、锦葵色素、芍药素(peonidin)、矮牵牛花色素(petunidin)和玫瑰色素(rosinidin)的糖苷。适用于营养组合物的这些和其它花青苷存在于多种植物源中。花青苷可以衍生自单一植物源或植物源的组合。适用于本发明组合物的富含花青苷的植物的非限制性实例包括:浆果(巴西莓(acai)、葡萄、越桔、蓝莓、越橘(lingonberry)、黑加仑子、阿龙尼亚苦味果、黑莓、覆盆子、樱桃、红加仑子、蔓越莓、红莓苔子、野生黄莓、欧洲越橘(whorleberry)、花楸浆果)、紫玉米、紫薯、紫萝卜、红甜薯、红卷心菜、茄子。
在一些实施方案中,本公开内容的营养组合物包含原花青素,其包括但不限于黄烷-3-醇和具有在2-11范围内的聚合度的黄烷-3-醇的聚合物(例如,儿茶素、表儿茶素)。这样的化合物可以衍生自单一植物源或植物源的组合。适用于所公开的营养组合物的富含原花青素的植物源的非限制性实例包括:葡萄、葡萄皮、葡萄籽、绿茶、红茶、苹果、松树皮、肉桂、可可、越橘(bilberry)、蔓越莓、黑加仑、阿龙尼亚苦味果。
适用于本发明营养组合物的黄烷-3-醇的非限制性实例包括儿茶素、表儿茶素、没食子儿茶素、表没食子儿茶素、表儿茶素没食子酸酯、表儿茶素-3-没食子酸酯、表没食子儿茶素和没食子酸酯。富含合适的黄烷-3-醇的植物包括,但不限于,茶、红葡萄、可可、绿茶、杏和苹果。
某些多酚化合物,特别是黄烷-3-醇,可通过增加脑血流(这与增加和持续脑能量/营养物递送以及新神经元的形成有关),改进人类受试者的学习和记忆。多酚也可提供神经保护作用且可增加大脑突触发生和抗氧化能力,从而支持年幼儿童的最佳脑发育。
用于营养组合物的优选的黄烷-3-醇源包括至少一种苹果提取物、至少一种葡萄籽提取物或其混合物。对于苹果提取物,黄烷-3-醇被分解成以4%-20%的范围存在的单体和以80%-96%的范围存在的聚合物。对于葡萄籽提取物,黄烷-3-醇被分解成总黄烷-3-醇和总多酚含量的单体(约46%)和聚合物(约54%)。聚合物黄烷-3-醇的优选的聚合度在约2和11之间的范围内。而且,苹果和葡萄籽提取物可含有儿茶素、表儿茶素、表没食子儿茶素、表儿茶素没食子酸酯、表没食子儿茶素没食子酸酯、聚合物原花青素、茋类(即白藜芦醇)、黄酮醇(即槲皮素、杨梅酮),或其任何混合物。富含黄烷-3-醇的植物源包括,但不限于苹果、葡萄籽、葡萄、葡萄皮、茶(绿茶或红茶)、松树皮、肉桂、可可、越橘(bilberry)、蔓越莓、黑加仑、阿龙尼亚苦味果。
如果将营养组合物给予儿科受试者,可以给予范围介于每日约0.01 mg和约450mg之间的量的黄烷-3-醇(包括单体黄烷-3-醇、聚合黄烷-3-醇或其组合)。在一些情况下,给予婴儿或儿童的黄烷-3-醇的量可在每日从约0.01 mg至约170 mg,每日从约50至约450mg,或每日从约100 mg至约300 mg的范围内。
在本公开内容的实施方案中,黄烷-3-醇以范围从约0.4至约3.8 mg/g营养组合物(约9-约90 mg/100 Kcal)的量存在于营养组合物中。在另一个实施方案中,黄烷-3-醇以范围从约0.8至约2.5 mg/g营养组合物(约20-约60 mg/100 Kcal)的量存在。
在一些实施方案中,本公开内容的营养组合物包含黄烷酮。合适的黄烷酮的非限制性实例包括紫铆亭、圣草酚、橙皮素(hesperetin)、橙皮苷、高北美圣草素、异樱花素、柚苷配基、柚皮素(naringin)、乔松素(pinocembrin)、枸橘素(poncirin)、樱花素(sakuranetin)、樱花甙(sakuranin)、steurbin。富含黄烷酮的植物源包括,但不限于橙子、橘子、葡萄柚、柠檬、酸橙。营养组合物可经配制以递送介于每日约0.01和约150 mg之间的黄烷酮。
此外,营养组合物还可包含黄酮醇。可以使用来自植物或藻类提取物的黄酮醇。黄酮醇,例如ishrhametin、山柰酚(kaempferol)、杨梅酮、槲皮素,可以足够递送给受试者介于每日约0.01和150 mg之间的量包含在营养组合物中。
营养组合物的植物营养素组分还可包含已在人乳中鉴定出的植物营养素,包括但不限于柚苷配基、橙皮素、花青苷、槲皮素、山柰酚、表儿茶素、表没食子儿茶素、表儿茶素-没食子酸酯、表没食子儿茶素-没食子酸酯或其任何组合。在某些实施方案中,营养组合物包含介于约50和约2000 nmol/L之间的表儿茶素,介于约40和约2000 nmol/L之间的表儿茶素没食子酸酯,介于约100和约4000 nmol/L之间的表没食子儿茶素没食子酸酯,介于约50和约2000 nmol/L之间的柚苷配基,介于约5和约500 nmol/L之间的山柰酚,介于约40和约4000 nmol/L之间的橙皮素,介于约25和约2000 nmol/L之间的花青苷,介于约25和约500nmol/L之间的槲皮素,或其混合物。而且,营养组合物可包含植物营养素或其母体化合物的代谢产物,或它可包含其它类型的膳食植物营养素,例如芥子油苷或萝卜硫素。
在某些实施方案中,营养组合物包含类胡萝卜素,例如叶黄素、玉米黄素、虾青素、番茄红素、β-胡萝卜素、α-胡萝卜素、γ-胡萝卜素和/或β-玉米黄质。富含类胡萝卜素的植物源包括,但不限于猕猴桃、葡萄、柑桔(citrus)、西红柿、西瓜、番木瓜和其它红色水果,或深绿色蔬菜,如羽衣甘蓝,菠菜,萝卜青菜(turnip greens)、宽叶羽衣甘蓝、罗马生菜(romainlettuce)、西兰花、绿皮密生西葫芦(zucchini)、豌豆和布鲁塞尔豆芽、菠菜、胡萝卜。
人不能合成类胡萝卜素,但在人类母乳中已鉴定出超过34种类胡萝卜素,包括某些类胡萝卜素的异构体和代谢产物。除了它们在母乳中的存在,膳食类胡萝卜素,例如α和β-胡萝卜素、番茄红素、叶黄素、玉米黄素、虾青素和玉米黄质也存在于喂乳母亲和母乳喂养的婴儿的血清中。类胡萝卜素一般被报道改善细胞与细胞之间的通信,促进免疫功能,支持健康的呼吸器官卫生,保护皮肤免于UV光的损害,并且与某些类型癌症和所有原因死亡率的风险降低相关。而且,膳食来源的类胡萝卜素和/或多酚由人受试者吸收,在母乳中积聚和保留,使得它们可为养育婴儿利用。因此,加入植物营养素至婴儿配方或儿童的产品中将使配方更接近人乳的组成和功能。
类黄酮,总体上,也可被包括在营养组合物中,因为类黄酮不能由人类合成。此外,来自植物或藻类提取物的类黄酮可以单体、二聚体和/或聚合物的形式使用。在一些实施方案中,营养组合物包含与哺乳期的头3个月期间人乳类似水平的单体形式的类黄酮。虽然类黄酮糖苷配基(单体)已在人乳样品中鉴定出,共轭形式的类黄酮和/或它们的代谢产物也可以用于营养组合物中。类黄酮可以下列形式加入:游离的、葡糖苷酸、甲基葡糖苷酸、硫酸酯和甲基硫酸酯。
营养组合物还可包含异类黄酮和/或异黄酮。实例包括,但不限于,木黄酮(染料木黄酮)、黄豆苷元(黄豆苷)、黄豆黄素(glycitein)、鹰嘴豆素A (biochanin A)、刺芒柄花素(formononetin)、考迈斯托醇(coumestrol)、德鸢尾素(irilone)、二羟四氢黄酮(orobol)、假靛黄素(pseudobaptigenin)、三尖叶猪异黄酮A和B (anagyroidisoflavone A和B)、毛蕊异黄酮(calycosin)、黄豆黄素、鸢尾苷元(irigenin)、5-O-甲基木黄酮、红车轴草素(pratensein)、樱黄素(prunetin)、Ψ-顶生配质(psi-tectorigenin)、凹猪屎豆碱(retusin)、鸢尾黄素(tectorigenin)、鸢尾定(iridin)、芒柄花苷(ononin)、葛根素、鸢尾苷(tectoridin)、大鱼藤异黄酮(derrubone)、羽扇豆异黄酮(luteone)、0,5,7,4'三羟基-6-异戊烯基异黄酮(wighteone)、猫尾草异黄酮(alpinumisoflavone)、吡喃异黄酮(barbigerone)、二-O-甲基猫尾草异黄酮和4'-甲基-猫尾草异黄酮。富含异类黄酮的植物源包括,但不限于,大豆、补骨脂、野葛、羽扇豆、蚕豆、鹰嘴豆(chickpea)、紫花苜蓿、豆荚和花生。营养组合物可经配制以递送介于每日约0.01和约150 mg之间的异黄酮和/或异类黄酮。
在一个实施方案中,本公开内容的营养组合物包含有效量的胆碱。胆碱是细胞正常功能必需的营养物。它是膜磷脂的前体,且它加速乙酰胆碱,一种涉及记忆存储的神经递质的合成和释放。而且,虽然不希望受到这种或任何其它理论的束缚,相信膳食胆碱和二十二碳六烯酸(DHA)协同作用促进磷脂酰胆碱的生物合成并因此帮助促进人类受试者中的突触发生。此外,胆碱和DHA可表现出促进树突棘形成的协同作用,这在建立的突触连接的维持中是重要的。在一些实施方案中,本公开内容的营养组合物包含有效量的胆碱,其是约20mg胆碱每8液量盎司(236.6 mL)份饮食-约100 mg每8液量盎司(236.6 mL)份饮食。
此外,在一些实施方案中,营养组合物是营养完全的,含有为受试者营养物的唯一来源的合适类型和量的脂质、碳水化合物、蛋白质、维生素和矿物质。事实上,营养组合物可任选地包含任何数目的蛋白质、肽、氨基酸、脂肪酸、益生菌和/或它们的代谢副产物、益生元、碳水化合物和可提供给受试者许多营养和生理学益处的任何其它营养物或其它化合物。此外,本公开内容的营养组合物可包含香料、增味剂、甜味剂、色素、维生素、矿物质、治疗成分、功能食物成分、食物成分、加工成分或其组合。
以下实施例描述了本公开内容的实施方案。考虑了本文公开的公开方法的说明书或实践,本文权利要求范围内的其它实施方案对于本领域技术人员而言将是显而易见的。意图是说明书连同实施例仅被认为是示例性的,本公开内容的范围和精神由实施例后面的权利要求书指示。在实施例中,除非另有说明,否则所有百分比均以重量计。
实施例1
在范围为50-5000μg/ml - 广泛水解酪蛋白(eHC)、> 500Da的广泛水解酪蛋白级分(eHC> 500Da)和氨基酸制备物(AA)的三种不同浓度下测试两种水解产物和一种氨基酸混合物。
在含有20mM 4-(2-羟乙基)-1-哌嗪乙烷磺酸(HEPES),0.01%辛基苯酚乙氧基化物(triton X-100)和2mM二硫苏糖醇(DTT) pH = 7.5,补充有4%二甲基亚砜(DMSO)的缓冲液中,将样品溶解为2%(20mg/mL)储液。在三种浓度50μg/mL,500μg/mL和5000μg/mL下,通过酶联免疫吸附测定(ELISA)、基于固定化金属离子亲和力的荧光偏振(IMAP)或迁移率变动测定(MSA)以KmATP测定激酶活性。
激酶数据在抑制热图中表示,包括与“+激酶”(100%)和“无激酶”(0%)相比的抑制百分比。使用以下公式计算%抑制:(1-(exp值-“无激酶”值)/(“+激酶”值-“无激酶”值))* 100。在热图中,负值设置为零(无抑制)。
表3
表4
以下趋势从表3和4可见:AA显示低活性,eHC显示对特定激酶的活性,并且eHC> 500Da比eHC更具活性,表明eHC中较长的序列最有可能介导激酶抑制。
激酶概况分析中存在明显的趋势。这种趋势与剂量有关,并且影响大多数测试的激酶。SYK和GSK3β是最有效被抑制的激酶。
AA在50μg/ml和500μg/ml不显示活性。在5000μg/ml时,在大多数激酶中可见中等活性(潜在的非特异性抑制)。
对于eHC和eHC> 500Da,在50μg/ml下SYK和GSK3β遵循该趋势。在500μg/ml下,PIM1, IKKβ, SYK, GSKβ, PDK1和AurA遵循该趋势。在5000μg/ml下,趋势遵循者是:生长因子受体激酶,特别是PDGFRα, EGFR, EPHA2, EPHB4, FLT3, HGK, KDR, SRC, TIE2和TRKA;炎症信号传导激酶,特别是IKKβ, IRAK4, JAK3, JNK2, LCK, p38α, SYK; 代谢信号传导,特别是GSK3β, PDK1, Erk2, SGK, 细胞周期激酶AurA; 和PIM1, PYK2, TSSK1。
对于eHC和eHC> 500Da,SYK和GSK3β在50μg/ml时被抑制。
TYRO3, CK1ε, DYRK1B, PKACα, PDK2和ROCK1在测试浓度下不被水解产物抑制。
eHC> 500Da比eHC更具活性,说明eHC的分级分离以剂量依赖性方式增加活性。
表5提供了根据本公开内容的营养组合物的示例性实施方案,并且描述了每100kcal份包括的每种成分的量。
表5. 实施例营养组合物的营养概况
虽然已使用特定术语、装置和方法描述本公开内容的优选实施方案,但是这类描述只用于说明目的。所用词汇是描述词汇而不是限制词汇。要了解,在不偏离随附权利要求书中阐述的本公开内容的精神或范围的情况下,本领域普通技术人员可进行变动和改变。另外,应了解,各个实施方案的各方面可全部或部分互换。因此,随附权利要求书的精神和范围不应限于其中所含的优选形式的描述。
本说明书引用的所有参考文献,包括而不限于所有的论文、出版物、专利、专利申请、简报、教科书、报告、手稿、小册子、书籍、互联网帖子、杂志文章、期刊等,藉此通过引用以其整体结合到本说明书中。本文的参考文献的讨论仅仅意在概括其作者所做出的断言,并非承认任何参考文献构成先有技术。申请人保留对所引用的参考文献的准确性和相关性提出异议的权利。
Claims (13)
1.通过给予受试者营养组合物而调节激酶的方法,所述营养组合物包含具有大于500道尔顿的摩尔质量分布的广泛水解酪蛋白、广泛水解酪蛋白级分或其组合。
2. 权利要求1的方法,其中所述营养组合物包含蛋白质等同物源,其中所述蛋白质等同物源的至少1%包含广泛水解酪蛋白、广泛水解酪蛋白级分或其组合,使得蛋白质等同物的至少1%至80%包含以下个体肽:SEQ ID NO 4, SEQ ID NO 13, SEQ ID NO 17, SEQ IDNO 21, SEQ ID NO 24, SEQ ID NO 30, SEQ ID NO 31, SEQ ID NO 32, SEQ ID NO 51,SEQ ID NO 57, SEQ ID NO 60和SEQ ID NO 63。
3.权利要求2的方法,其中所述蛋白质等同物源以约0.2g/100kcal至约5.6g/100kcal的营养组合物的量存在。
4. 权利要求2的方法,其中所述蛋白质等同物源还包含选自以下的至少10种个体肽:SEQ ID NO 1, SEQ ID NO 2, SEQ ID NO 3, SEQ ID NO 5, SEQ ID NO 6, SEQ ID NO 7,SEQ ID NO 8, SEQ ID NO 9, SEQ ID NO 10, SEQ ID NO 11, SEQ ID NO 12, SEQ ID NO14, SEQ ID NO 15, SEQ ID NO 16, SEQ ID NO 18, SEQ ID NO 19, SEQ ID NO 20, SEQID NO 22 , SEQ ID NO 23, SEQ ID NO 25, SEQ ID NO 26, SEQ ID NO 27, SEQ ID NO28, SEQ ID NO 29, SEQ ID NO 33, SEQ ID NO 34, SEQ ID NO 35, SEQ ID NO 36, SEQID NO 37, SEQ ID NO 38, SEQ ID NO 39, SEQ ID NO 40, SEQ ID NO 41, SEQ ID NO42, SEQ ID NO 43, SEQ ID NO 44, SEQ ID NO 45, SEQ ID NO 46, SEQ ID NO 47, SEQID NO 48, SEQ ID NO 49, SEQ ID NO 50, SEQ ID NO 52, SEQ ID NO 53, SEQ ID NO54, SEQ ID NO 55, SEQ ID NO 56, SEQ ID NO 58, SEQ ID NO 59, SEQ ID NO 61, SEQID NO 62, SEQ ID NO 64及其组合。
5.权利要求1的方法,其中所述营养组合物还包含至少一种长链多不饱和脂肪酸。
6.权利要求5的方法,其中所述至少一种长链多不饱和脂肪酸选自二十二碳六烯酸和花生四烯酸。
7.权利要求1的方法,其中所述营养组合物还包含来自益生菌分批培养过程的指数生长后期的培养物上清液。
8.权利要求1的方法,所述营养组合物还包含益生菌。
9.权利要求1的方法,所述营养组合物还包含益生元。
10.权利要求9的方法,其中益生元组分包含聚葡萄糖。
11.权利要求10的方法,其中益生元组分还包含寡聚半乳糖。
12.权利要求1的方法,其中所述营养组合物还包含脂肪源和碳水化合物源。
13.权利要求1的方法,其中所述营养组合物是婴儿配方。
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