CN1330517A - 蛋黄中的食物诱导的抗分泌性蛋白 - Google Patents
蛋黄中的食物诱导的抗分泌性蛋白 Download PDFInfo
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Abstract
本发明总体上涉及天然抗分泌性蛋白(NASP)在制备富含NASP的功能性食物产品或者用于治疗和预防由极端体液排出而引起的疾病样状态的药物产品中的应用。特别是,本发明涉及富含NASP的蛋黄作为NASP源的应用。
Description
本发明总体上涉及天然抗分泌性蛋白(NASP)在制备富含NASP的功能性食物产品和用于治疗和预防由特别高水平的体液排出而引起的异常生理状态的药物产品中的应用。特别是,本发明涉及富集了NASP的蛋黄在制备这类产品中的应用。本发明还涉及这样制备的食物产品和药物产品及其制备方法。
下文中将使用术语“NASP”表示“天然抗分泌性蛋白”,而使用术语“ASP”表示包括在天然抗分泌性蛋白之内的“抗分泌性蛋白”。
从瑞典专利SE9000028-2(公布号466,331)得知,通过向动物饲料中添加某些糖、氨基酸和酰胺可以刺激抗分泌性因子或抗分泌性蛋白(ASP:也称FIL)的形成。要用于形成目的量ASP的这些物质的种类和用量是通过该专利中公开的一种方法确定的。简言之,该方法涉及测定大鼠小肠中的标准化分泌应答。由该专利显而易见的是,所形成的被诱导的ASPs指导体液分泌到肠。因此,腹泻状况可得以预防或治愈,这意味着更安全地生产动物产品。该专利中,天然抗分泌性蛋白的含量或用量根据其对进入实验室大鼠(它们已经用霍乱毒素激发过)的小肠的流体分泌作用的效果而确定。一个ASP单位(FIL单位)相当于与没有ASP的对照相比肠中流体流量降低50%。极少量的抗分泌性蛋白就有活性,因此通过它们的效果比通过它们的质量更容易测定它们。
从PCT申请PCT/SE96/01049得知,某些抗分泌性蛋白的结构及其活性部位得到了鉴定。已经表明,一种由重组遗传工程或由固相技术制备、具有确定的结构的合成ASP对活细胞膜上的体液流动有总体的控制效果。
从PCT申请PCT/SE97/01918(WO98/21978)得知,食用某种具有酶活性的食物可在体内诱导ASP的形成。诱导作用的效果以及由此导致的ASP的形成根据个体及其症状而不同,并且以目前不可预测的强度和诱导期间而发生。然而,可在以后测定它们,而且可在所述测定结果的指导下进行必要的校正。
在本发明人按照瑞典专利SE9000028-2调制饲料以刺激抗分泌性蛋白的形成的长期工作中,出乎意料地发现了,所形成的天然抗分泌性蛋白的合成能力和富集情况在体内的分布很不均匀。在某些器官、身体部位或体液内发现了很高水平的NASP,特别是在禽类蛋的蛋黄内。本发明基于这一出乎意料的发现,因而涉及富集了NASP的蛋黄在制备富含NASP的食物产品和用于治疗和预防由极端体液排出而引起的异常生理状态的药物产品中的应用。因此,本发明开辟了目前尚不知道的通过食用原样的蛋黄或呈食物产品形式的蛋黄而将NASP直接引入食物或饲料的可能性。
本发明总体上涉及天然抗分泌性蛋白在制备富含NASP的功能性食物产品和用于治疗和预防由极端体液排出而引起的异常生理状态的药物产品中的应用。本发明的一个方面涉及富集了NASP的蛋黄在制备具有高水平NASP的食物产品中的应用。本发明的另一方面涉及它们在制备特殊营养目的的食物和含有NASP的药物产品中的应用。
在本申请的上下文中,“功能性食物产品”是指具有有益于健康的功能(即,对人或动物的健康具有有益效果)的食物产品。
在本申请的上下文中,“食物产品”这一表述旨在包括供人使用的食物以及供动物使用的饲料。所述食物可以是产品形式的物品,其制品包括蛋黄,例如,薄烤饼、煎蛋饼、冰淇淋以及各种面包。该蛋黄还可以蛋黄乳或者煮老的蛋或嫩煮的蛋的形式食用。所述食物还可以呈面包、饼干、面食制品、酒糟和薄片、麦片粥或薄糊的形式,或者混入各种含有或不含电解质的饮料组合物,或含有肉和肉制品的食物制品,根据下述本发明一个实施方案的富集了NASP的脂肪和脂肪产品或者乳和乳产品,根据该方案,NASP已经以或多或少纯化的形式混入食物产品中。为了符合有关可口性和膳食变化性(meal variation)的要求,可应用技术人员已知的知识以极大的自由度对食物制品进行调配。
所述的蛋黄来源于可供人和动物食用并富集了NASP的禽类的蛋。这些蛋优选是由产卵的母鸡生产的,但也可由例如鹌鹑和鸵鸟获得。
“富含NASP的食物产品”或“具有高水平的NASP的食物产品”是指含有一定浓度的NASP的食物产品,该浓度使得食用该食物产品时能对健康提供所需的有价值的效果。正如由实施例1可见的那样,在蛋中可测定到特别高水平的NASP。当食用富含NASP的产品时,要设法使血中的ASP水平超过0.5FIL单位/ml血。健康人血中ASP水平似乎在0和0.5FIL单位/ml血之间。
按SE9000028-2刺激NASP的产生时,已经惊人而出乎意料地表明,在禽类的蛋中可诱导极高水平的NASP。经过这种诱导之后,血中的NASP水平通常为0.5-2FIL单位/ml血(1FIL单位=在对大鼠进行的所谓肠环试验(intestine loop test)中对流体排出具有50%抑制作用的NASP的量;参见SE9000028-2和其中引用的参考文献3和4)。按SE9000028-2,通过经饲料给禽类提供氨基酸、糖和酰胺的独特组合,可以优选在蛋黄中积累很高水平的NASP。由此提供了高达约1,000-约10,000FIL单位/ml的水平。在该上下文中应提到的是,在没有前面的诱导作用的情况下,蛋黄中的NASP水平通常约为1FIL单位/ml;参见S.Lange等人,British Poultry Science(1994)35:615-620。
根据本发明的用途的一个实施方案,可通过将NASP原样混入食物中而制备富含NASP的食物产品,其中所述的NASP是由富含NASP的蛋黄以或多或少纯化的形式获得的。
于是,可能如SE9000028-2中公开的那样刺激禽类中NASP的形成,然后还如所述瑞典专利公开的那样在琼脂糖柱上从蛋黄消化物中回收或浓缩NASP,接着用α-甲基糖苷在生理盐水(优选是磷酸盐缓冲过的)中的溶液洗脱它。然后,在例如以溶液形式(sol)施用之前,可对该洗脱物进行进一步的纯化和调制。
在这种情况下,随后可以将回收或浓缩的NASP施用给动物或人,分别与饲料或食物混合,或者作为或多或少分离的产品,制备和调制成药物或其它有利健康的产品。这类药物产品是以技术人员已知的方式、使用包括载体和稀释剂在内的被接受的赋形剂制备的,并且根据所需的施用途径调制成固体或液体形式。
根据本发明的制备富含NASP的食物产品的用途的一个实施方案,使用原样的蛋黄或使用完整的蛋,可能是经过加工的形式,用于与食物或饲料混合。任意的加工可包括研磨、浸提、萃取、蒸发、超滤、干燥以及类似的标准操作以获得一种适合实际目的的、富含NASP的蛋黄产品,用于与食物或饲料混合。
根据本发明,开辟了实现与NASP摄入相关的有利效果的新途径。鉴于可诱导的在蛋黄内的高浓度的NASP,NASP的效果可得以利用,而无须等待先前已知的NASP诱导作用引起的时间延迟。
于是,本发明的又一方面涉及一种从禽类的蛋提供蛋黄的方法,所述禽类的蛋富集了NASP,该方法的特征在于,按SE9000028-2用诱导NASP的饲料喂养要从其取蛋的禽类而刺激NASP的形成。特别是,饲养鹑鸡类的禽类时,本发明的方法是可适用的,因为如上述的那样,已经惊人而出乎意料地表明,在所述禽类的蛋中可诱导极高水平的NASP。同样如上述的那样,通过经饲料给雌禽鸡提供氨基酸、糖和酰胺的独特组合,可以优选在蛋黄中积累很高水平的NASP(大约100-1,000FIL单位)。因此,开辟了按照该专利中公开的发明制备具有极高水平的NASP的蛋黄的特殊的可能性。通过将这种蛋黄或其产品混入食物或饲料中,可以一种方便、安全而经济的方式将NASP补充给人或动物。于是,非常惊人而且先前不为人知的是,可以在食物中获得如此高水平的NASP。因而,如上述的那样,NASP的效果可得以利用,而无须等待先前已知的NASP诱导作用引起的时间延迟。同样出乎意料的是,即使口服时,NASP也保持其效果。
在某些情况下,可能需要高剂量的NASP。在这些情况下,可将由蛋回收的NASP添加到食物产品中,或者,可将NASP制成供摄入的片剂或悬浮液的形式。
一个特别优选的实施方案包括用喷雾干燥法制备蛋黄粉。这种产品在各种食物产品的工业加工中是特别适合的,在所述工业加工中,希望摄入该食物产品时能获得NASP的有益效果。该蛋黄粉使其自身能很好地混入象香肠和冰淇淋这样差别极大的产品中,正因为如此,才有可能实现在膳食特性(meal character)方面的大的变化。
在希望通过浸提或萃取制备富集或浓缩的NASP产品的情况下,蛋黄粉同样最适合作为中间体,用于以后制备例如药物。
由于其较低的分子量,NASP较为热稳定,于是在制备供食用的食物之前就可以含于其中。
已经表明,NASP对活细胞膜上的体液流动有总体的控制效果,因此,不仅腹泻状况能得以缓解、治愈或预防,而且由生理失调导致的不适或由极端体液排出而引起的疾病样状态(例如炎症、浮肿、关节炎、青光眼)以及体内的其它变化(例如偏头痛、灼伤和外伤性损伤)也是如此。于是,本发明的食物产品和药物产品可用于该目的。
下面通过如下的非限定性实施例阐释本发明。
实施例1
用按SE9000028-2配制的产蛋母鸡饲料和常规类型的对照饲料(11.4MJ/kg饲料,170g粗蛋白/kg饲料)饲养产蛋的母鸡(约32周龄的白色来航鸡)。经两周的诱导期之后,收集鸡蛋。
从每组收集10只鸡蛋,将这些鸡蛋的蛋黄和蛋清手工分离。用同样大体积的PBS(磷酸盐缓冲盐水)稀释后,按前述方法对蛋黄进行亲合色谱处理(I.Lnnroth和S.Lange,Biochimica Biophysica Acta(1986)883:138-144)。简言之,将PBS稀释过的蛋黄加样到已经用PBS预平衡过的含有 “Sepharose 6B”(Pharmacia LKB BiotechnologyAB,Sollentuna,Sweden)的小柱上。用PBS洗涤后,用甲基-α-D-葡糖苷在PBS中的1.0M溶液洗脱NASP。对PBS浸析后,将样品于-20EC下贮藏,直至按Lange描述的对大鼠的肠环模型进行有关NASP的试验(S.Lange,FEMS Microbiology Letters(1982)15:239,242)。
蛋黄中的NASP水平 对照组 试验组
(FIL单位/g) 1.20 2.0(稀释1∶100时)
1.0(稀释1∶1,000时)
实施例2
一名42岁、经过造瘘术、残留约为0.5m的妇女患有严重的腹泻。平均而言,每天4次分泌大约1.5升内脏内含物。经过一周的每日摄入由糖、奶油和实施例1的蛋黄制成的冰淇淋(相当于每日摄49干蛋黄物质)后,排粪频率减少到每日一次,排粪量减少到大约350ml。在三周的治疗期间,该妇女的体重增加了3.5kgs。又过了一周后,她食用相同量的按相同配方由普通蛋制备的冰淇淋。三天之内,排粪频率和排粪量达到与该试验前相同的值。又过了一周后,继续进行该试验,在两天之内,排粪减少到与前次试验期间相同的值。
实施例3
将四头鲜重50kgs的小牛分成两组,每组有两头小牛。给对照组的小牛提供商品乳代用品形式的小牛营养物。在试验组中,提供得自实施例1实验的混合蛋黄,相当于两个蛋黄的日剂量。饲养持续七天。在实验前和实验后测定血中的ASP水平。
ASP水平
(FIL单位/ml血浆)
对照组 试验组
实验前 0.3 0.1
实验 0.1 0.9
实施例4
腹泻是狗类中的常见病,特别是那些在乡村饲养的有可能自主择食的狗。这样的一只德国牧羊狗(1.5岁龄、体重约40kgs)一段时间来患有严重腹泻和带水的、周期性的带血排粪。这只狗还间歇性地呕吐。看过兽医,开出抗生素处方,口服片剂一周。在此期间,这只狗吃较少量的狗用商品干饲料。没有发现腹泻的减少。该抗生素治疗后,用浸泡过的干饲料(其中有得自实施例6的蛋黄粉,用量相当于每天一个蛋黄)喂狗四天。一昼夜后,这只狗有了固形排粪,再过三天后,认为它已经很健康了。在此以前,狗主人从未见过这样快的恢复。
实施例5
将得自实施例1中试验组的蛋打碎,在蛋分离装置中分离。各自分别收集蛋黄和蛋清。在一个“Anhydro”喷雾干燥器中将蛋黄喷雾干燥,入口温度为150EC,出口温度约为75EC。
收集蛋粉,完全干燥后,收集粘在喷雾干燥器壁上的蛋粉。这些壁沉积物在干燥室中的平均停留时间为4小时。将蛋粉溶于水,按公知方式在琼脂糖柱上收集ASP。将洗脱物稀释,测定其ASP含量。
稀释1∶100后的FIL单位
蛋粉,平均样品 X
得自干燥室壁的蛋粉 Y
因此,从食品工艺的角度来看,NASP是足够热稳定的。
技术效果
本发明以全新、先前未知的独立性程度,提供了施用NASP的全新、先前未知的可能性。PCT申请PCT/SE97/01918(WO98/21978)中公开的NASP诱导作用可能难于长期保持,这归因于饮食更容易被认为单调这一事实。通过本发现,可以适度的加工成本制备高可口性和良好的接受性的食物产品。在长病程或对诱导NASP的食物的应答弱的情况下,可施用食物形式的NASP,该食物具有适合于对生活质量或身体性能效率的所需效果的、预定的或高或低的剂量。在长期施用的情况下,可避免不喜欢(厌恶)食物,因为可以制备具有不同味道和膳食特性的许多种产品。可用工业方法将食物产品制成半制成的食物或制成的膳食或者制成适用作制备药物中的或生产NASP浓缩物中的粗产品的物品。可以通过萃取或浸提从蛋黄或优选从蛋黄粉回收浓缩物。
Claims (24)
1.具有高水平天然抗分泌性蛋白(NASP)的蛋黄。
2.权利要求1的蛋黄,其NASP含量相当于1,000—10,000FIL单位/ml。
3.权利要求1或2的蛋黄,它来源于鹑鸡类的禽类的蛋。
4.权利要求1—3任一项的蛋黄在制备富含NASP的功能性食物产品中的应用。
5.权利要求1—3任一项的蛋黄在制备用于治疗和预防由极端体液排出而引起的疾病样状态的药物产品中的应用。
6.NASP本身根据权利要求4或5的应用,所述NASP以纯化或浓缩的形式回收自富含NASP的蛋黄。
7.权利要求4或5的应用,其特征在于,所述蛋黄照原样应用或者以一种通过加工获得的形式应用,所述加工例如是研磨、浸提、萃取、蒸发、膜过滤、干燥以及类似的标准操作。
8.权利要求4—7任一项的应用,其特征在于,通过用诱导NASP的饲料喂养产蛋的禽类而提供了所述蛋黄。
9.权利要求8的应用,其特征在于,通过将氨基酸、糖和酰胺混入所述饲料而实现所述NASP诱导作用,混入的量和比例使NASP的形成得到刺激。
10.权利要求8的应用,其特征在于,通过将具有刺激NASP的形成的酶活性的产物混入所述饲料而实现所述NASP诱导作用。
11.权利要求10的应用,其特征在于,所述具有酶活性的产物是发芽的谷类。
12.一种从产蛋的禽类提供蛋黄的方法,所述蛋黄富集了天然抗分泌性蛋白(NASP),其特征在于,用诱导NASP的饲料喂养所述的禽类。
13.权利要求12的方法,其特征在于,所述禽类是一种鹑鸡类的禽类。
14.权利要求12或13的方法,其特征在于,所述诱导NASP的饲料含有含量能刺激NASP的形成的氨基酸、糖和酰胺。
15.权利要求12或13的方法,其特征在于,所述饲料含有具有刺激NASP的形成的酶活性的产物。
16.权利要求15的方法,其特征在于,所述具有酶活性的产物是发芽的谷类。
17.具有高水平天然抗分泌性蛋白(NASP)的食物产品,其特征在于,含有权利要求1—3任一项的蛋黄或者得自蛋黄的或多或少纯化形式的NASP。
18.权利要求17的食物产品,其特征在于,通过在制备该食物产品时往其中混入作为NASP源的、权利要求1—3任一项的富含NASP的蛋黄或其加工形式而获得了高水平的NASP。
19.权利要求17的食物产品,其特征在于,通过在制备该食物产品时往其中添加纯化或浓缩形式的NASP本身而获得了高水平的NASP,所述纯化或浓缩形式的NASP本身得自权利要求1—3任一项的富含NASP的蛋黄。
20.权利要求17的食物产品,其特征在于,它包括蛋黄本身。
21.权利要求17—19任一项的食物产品,其特征在于,它包括薄烤饼、煎蛋饼、冰淇淋、煮老的蛋或嫩煮的蛋等等。
22.用于治疗和预防由极端体液排出而引起的疾病样状态的药物产品,其特征在于,除了常规赋形剂和辅助剂之外,还含有来源于权利要求1—3任一项的富含NASP的蛋黄的天然抗分泌性蛋白(NASP)。
23.一种制备权利要求17—21任一项的具有高水平天然抗分泌性蛋白(NASP)的食物产品的方法,其特征在于,在食物或饲料中加入权利要求1—3任一项的富含NASP的蛋黄或其加工形式或得自蛋黄的或多或少纯化的NASP。
24.权利要求23的方法,其特征在于,通过研磨、浸提、萃取、蒸发、膜过滤、干燥以及类似的标准操作获得了蛋黄的加工形式。
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EP4125984A1 (en) | 2020-03-26 | 2023-02-08 | Lantmännen Functional Foods AB | A consumable product comprising malted cereals for promoting recovery at physical activity |
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Publication number | Priority date | Publication date | Assignee | Title |
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SE466331B (sv) | 1990-01-04 | 1992-02-03 | Svenska Lantmaennens | Saett att korrigera och optimera naeringssammansaettningen i foeda |
US5464637A (en) * | 1992-02-27 | 1995-11-07 | C. Itoh Feed Mills Co., Ltd. | Egg containing iron or vitamin D3 in high content |
SE508609C2 (sv) | 1995-08-24 | 1998-10-19 | Rural Patent Svenska Ab | Antisekretorisk faktor - dess aminosyrasekvens, nubleinsyrasekvens och användning |
SE9604251L (sv) * | 1996-11-20 | 1997-12-22 | Svenska Lantmaennen Riksfoerbu | Födoämne som vid förtäring inducerar antisekretoriska proteiner |
SE508086C2 (sv) * | 1996-12-16 | 1998-08-24 | Valmet Karlstad Ab | Rullstol |
SE513496C2 (sv) * | 1998-12-17 | 2000-09-18 | Rural Patent Svenska Ab | NASP-berikad äggula samt dess användning |
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1998
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101437533B (zh) * | 2006-04-27 | 2016-10-12 | 兰特门内阿斯-法克托尔公司 | 抗分泌蛋白对于治疗间室综合症的应用 |
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