TWI626893B - Method for inhibiting pathogens using a nutritional composition - Google Patents
Method for inhibiting pathogens using a nutritional composition Download PDFInfo
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- TWI626893B TWI626893B TW100148650A TW100148650A TWI626893B TW I626893 B TWI626893 B TW I626893B TW 100148650 A TW100148650 A TW 100148650A TW 100148650 A TW100148650 A TW 100148650A TW I626893 B TWI626893 B TW I626893B
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- Prior art keywords
- lactoferrin
- kcal
- human
- source
- nutritional composition
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Abstract
一種用於抑制細菌入侵或附著之機轉的方法,其涉及對人投予營養組成物,該營養組成物包括脂質或脂肪;蛋白質來源;益菌生組成物;以及至少約10 mg/100kCal的藉由非人來源所製造之乳鐵蛋白。 A method for inhibiting the mechanism of bacterial invasion or attachment, which involves administering to a human a nutritional composition including lipids or fats; a source of protein; a probiotic composition; and at least about 10 mg / 100kCal Lactoferrin made from non-human sources.
Description
本文大致上關於例如嬰兒配方、人乳強化營養品、孩童膳食補充品等等之營養組成物領域,該營養組成物中具有乳鐵蛋白。更特定言之,本文係關於一種,藉由投予人包括由非人來源所製造之乳鐵蛋白的營養組成物來抑制有害細菌病原體之入侵機轉和/或抑制在人胃腸道中至少一種病原體的附著之方法。 This document relates generally to the field of nutritional compositions such as infant formula, human milk fortified supplements, children's dietary supplements, etc., which have lactoferrin in the nutritional composition. More specifically, this article relates to a method for inhibiting the invasion of harmful bacterial pathogens and / or inhibiting at least one pathogen in the human gastrointestinal tract by administering to a human a nutritional composition including lactoferrin produced by non-human sources Method of attachment.
目前有各種針對人、特別針對年輕人的膳食組成物,以提供在某些生命階段的補充性或主要營養。一般而言,市售針對嬰兒的膳食組成物盡可能模擬人乳的組成物及相關功能性之程度。透過組合其中某些具有生理活性的蛋白質及經摻合脂肪成分,調製膳食組成物以使彼等模擬人乳,以用作為完整或部分取代物。其他常利用於嬰兒膳食組成物中的成分可包括例如乳糖之碳水化合物來源和咸信存在於人乳中以供嬰兒吸收的維生素、礦物質及元素。 There are various dietary compositions for humans, especially young people, to provide supplemental or primary nutrition during certain life stages. In general, commercially available dietary compositions for infants mimic the composition of human milk and related functionalities as much as possible. By combining some of these proteins with physiological activity and blended fat ingredients, the dietary composition is modulated to make them mimic human milk for use as complete or partial substitutes. Other ingredients commonly used in infant dietary compositions may include, for example, carbohydrate sources of lactose and vitamins, minerals and elements that are present in human milk for absorption by infants.
一些存在於人乳中的蛋白質提供對抗病原體之防禦,以預防及抑制感染,另外同時促進嬰兒體內免疫反應。包括咕啉結合蛋白(haptocorrin,維生素B12結合蛋白)及乳鐵蛋白的蛋白質被認為能經由抑菌及殺菌活性幫助嬰兒防禦抵抗各種細菌病原體。 Some of the proteins found in human milk provide defense against pathogens to prevent and suppress infections, while also boosting the immune response in infants. Proteins including haptocorrin (vitamin B12 binding protein) and lactoferrin are believed to help infants defend against various bacterial pathogens through bacteriostatic and bactericidal activities.
乳鐵蛋白係人乳中的主要蛋白質之一,且被認為是具 有平均分子量約80千道耳頓的醣蛋白。其為鐵結合蛋白,具有以可逆形式結合二個鐵分子的能力,且可促進在人的小腸內之鐵攝取。功能上,乳鐵蛋白調控鐵吸收且由此可結合鐵基自由基,並供予鐵以供免疫反應之用。 Lactoferrin is one of the main proteins in human milk and is considered to have There are glycoproteins with an average molecular weight of about 80 kilodaltons. It is an iron-binding protein that has the ability to bind two iron molecules in a reversible form and can promote iron uptake in the human small intestine. Functionally, lactoferrin regulates iron absorption and can thus bind iron-based free radicals and supply iron for immune responses.
通常,乳鐵蛋白的另外角色係其保衛人的小腸感染方面之抗微生物活性,尤其是針對嬰兒。已知乳鐵蛋白在特定細菌生長抑制之同時,在成功入侵小腸細胞前殺除微生物的抑菌性及殺菌性。 Often, the additional role of lactoferrin is in its antimicrobial activity in defending human intestinal infections, especially in infants. Lactoferrin is known to inhibit the bacteriostatic and bactericidal properties of microorganisms before successfully invading small intestinal cells while inhibiting the growth of specific bacteria.
為獲得商業可行性之膳食組成物,加入乳鐵蛋白通常受到侷限,此乃由於在加工處理期間預期失去活性所致。舉例而言,通常在加工處理嬰兒配方及其他例如人乳強化營養品及各種孩童產品中的溫度及pH要求而降低了乳鐵蛋白的特定功能,造成最終調製物中未含乳鐵蛋白。此外,針對乳鐵蛋白常僅考慮到其鐵結合性;因此,在此種性質被認為藉由加工處理條件而變少時,調製物通常可能不具乳鐵蛋白。 In order to obtain a commercially viable dietary composition, the addition of lactoferrin is often limited due to the expected loss of activity during processing. For example, the temperature and pH requirements of infant formulas and other fortified human milk supplements and various children's products often reduce the specific functions of lactoferrin, resulting in the absence of lactoferrin in the final preparation. In addition, lactoferrin is often considered only for its iron-binding properties; therefore, when this property is considered to be reduced by processing conditions, the modulator may generally not have lactoferrin.
又,如本技術領域所知,人乳的鐵含量相對較低(每升人乳含有約0.3毫克鐵)。雖然含量低,但人類嬰兒具有高吸收率,吸收人乳中約一半的鐵。然而,當餵予人類嬰兒具有例如每升約10 mg至約12 mg的高量鐵營養強化之先前技術配方時,嬰兒吸收少於總鐵量的約5%。以該先前技術配方中的高鐵量,雖然乳鐵蛋白係已知的鐵運輸蛋白,基本上所有乳鐵蛋白鐵結合位置預期均會被占據。 Also, as is known in the art, human milk has a relatively low iron content (containing about 0.3 mg of iron per liter of human milk). Although low in content, human infants have high absorption rates, absorbing about half of the iron in human milk. However, when human infants are fed with prior art formulations with high levels of iron nutrition fortification, such as about 10 mg to about 12 mg per liter, the infant absorbs less than about 5% of the total iron content. With the high iron content in this prior art formulation, although lactoferrin is a known iron transporter, basically all lactoferrin iron binding sites are expected to be occupied.
該先前技術配方的其他困難包括無能力提供抑菌作用 。這一部分係由於使用具有經阻斷或已損壞之結合位置之乳鐵蛋白,然而抑菌作用至少部分相關於存在於配方中的乳鐵蛋白之鐵結合程度。 Other difficulties with this prior art formulation include the inability to provide bacteriostatic effects . This is partly due to the use of lactoferrin with a blocked or damaged binding site, but the bacteriostatic effect is at least partially related to the degree of iron binding of the lactoferrin present in the formulation.
因此,有益的是,提供例如嬰兒配方、人乳營養強化品、孩童的膳食補充等等含有乳鐵蛋白、特別是藉由非人來源製造的乳鐵蛋白之營養組成物。較佳的是,包括在該組成物中的乳鐵蛋白即使於高溫及低pH條件下加工處理後仍具有抑菌效果。結合包括經過例如於巴斯德殺菌期間之高或低pH或高溫條件而維持乳鐵蛋白抗入侵或抗附著機轉之特性,提供可至少部分保護抵抗有害細菌病原體的膳食組成物。 Therefore, it would be beneficial to provide nutritional compositions containing lactoferrin, especially lactoferrin made from non-human sources, such as infant formula, human milk nutrition supplements, dietary supplements for children, and the like. Preferably, the lactoferrin included in the composition has a bacteriostatic effect even after processing under high temperature and low pH conditions. The combination includes maintaining lactoferrin's resistance to invasion or resistance to mechanical attachment through high or low pH or high temperature conditions, such as during pasteurization, to provide a dietary composition that can at least partially protect against harmful bacterial pathogens.
簡言之,本文在一實施例中係關於一種使用包含脂質或脂肪、蛋白質來源、益菌生組成物及來自非人來源之乳鐵蛋白的營養組成物以用於抑制一或多種細菌病原體入侵機轉和/或至少一種病原體的附著之方法,其中該組成物即使於包括暴露於嚴苛環境條件的加工處理後仍提供抵抗人胃腸道內不希望之細菌菌株的活性抗入侵機轉。 Briefly, in one embodiment, the present invention relates to the use of a nutritional composition comprising lipid or fat, a protein source, a probiotic composition, and lactoferrin from a non-human source for inhibiting the invasion of one or more bacterial pathogens. A method of mechanical transformation and / or attachment of at least one pathogen, wherein the composition provides an active anti-invasive mechanism against undesired bacterial strains in the human gastrointestinal tract even after processing including exposure to harsh environmental conditions.
在一實施例中,本文係關於一種營養產品,其包含:a.至高達約7 g/100 kcal的脂肪或脂質,更佳為約3 g/100 kcal至約7 g/100 kcal的脂肪或脂質;b.至高達約5 g/100 kcal的蛋白質來源,更佳為約1 g/100 kcal至約5 g/100 kcal的蛋白質來源;c.至少約10 mg/100 kcal的乳鐵蛋白,更佳為約70 mg/100 kcal至約220 mg/100 kcal藉由非人來源所製造之乳鐵蛋白,最佳為約90 mg/100 kcal至約190 mg/100 kcal藉由非人來源所製造之乳鐵蛋白;以及d.約0.1 g/100 kcal至約1 g/100 kcal之益菌生組成物,其包含聚葡萄糖(polydextrose)和/或半乳糖寡醣。更佳的是,該營養組成物包含約0.3 g/100 kcal至約0.7 g/100 kcal的益菌生組成物,其包含聚葡萄糖及半乳糖寡醣之組合。 In one embodiment, this document relates to a nutritional product comprising: a. Up to about 7 g / 100 kcal of fat or lipid, more preferably about 3 g / 100 kcal to about 7 g / 100 kcal of fat or Lipids; b. Protein sources up to about 5 g / 100 kcal, more preferably about 1 g / 100 kcal to about 5 g / 100 kcal of protein source; c. lactoferrin of at least about 10 mg / 100 kcal, more preferably about 70 mg / 100 kcal to about 220 mg / 100 kcal by non-human sources The lactoferrin produced is preferably about 90 mg / 100 kcal to about 190 mg / 100 kcal, lactoferrin manufactured from non-human sources; and d. About 0.1 g / 100 kcal to about 1 g / 100 A probiotic composition of kcal, which comprises polydextrose and / or galactooligosaccharide. More preferably, the nutritional composition comprises a probiotic composition of about 0.3 g / 100 kcal to about 0.7 g / 100 kcal, which comprises a combination of polydextrose and galactooligosaccharide.
較佳的是,該乳鐵蛋白係非人乳鐵蛋白和/或藉由經基因修飾之生物體所製造之人乳鐵蛋白。術語「生物體」如本文中所用,指任何持續之生命系統,例如動物、植物、真菌或微生物。在一特佳實施例中,所使用之乳鐵蛋白係使有效量之含乳鐵蛋白營養組成物可被投予,以抑制人胃腸道中至少一種病原體之入侵機轉之至少一者,即使於加工處理期間,該營養組成物已經暴露於典型的某些類似巴斯德殺菌之加工處理條件的pH及溫度波動亦然。該乳鐵蛋白包括可破壞某些細菌於人體細胞上所使用之附著因子及注射針(injection needle)的抗入侵機轉。在另一特佳實施例中,所使用之乳鐵蛋白係使有效量之含乳鐵蛋白營養組成物可被投予,以抑制人胃腸道中至少一種病原體之附著,即使於加工處理期間,該營養組成物已經暴露於典型的某些類似巴斯德殺菌之加工處理條件的pH及溫度 波動亦然。此種病原體實例包括腸毒性大腸桿菌(ETEC)、腸內病原性大腸桿菌(EPEC)、流行性感冒嗜血桿菌(Haemophilus influenza)、產志賀毒素性大腸桿菌(STEC)、腸凝集性大腸桿菌(EAEC)、鼠傷寒血清沙氏桿菌血清型(Salmonella ser.Typhimurium)、副痢疾桿菌(Shigella flexneri)、輪狀病毒、諾羅病毒、呼吸道融合性病毒(RSV)、腺病毒及其組合。 Preferably, the lactoferrin is a non-human lactoferrin and / or a human lactoferrin produced by a genetically modified organism. The term "organism" as used herein refers to any continuous living system, such as an animal, plant, fungus, or microorganism. In a particularly preferred embodiment, the lactoferrin is used so that an effective amount of a lactoferrin-containing nutritional composition can be administered to inhibit at least one of the invasion mechanisms of at least one pathogen in the human gastrointestinal tract, even if During processing, the nutritional composition has been exposed to pH and temperature fluctuations typical of some pasteurized processing conditions. The lactoferrin includes an anti-invasion mechanism that can destroy the adhesion factors and injection needles used by certain bacteria on human cells. In another particularly preferred embodiment, the lactoferrin is used so that an effective amount of a lactoferrin-containing nutritional composition can be administered to inhibit the adhesion of at least one pathogen in the human gastrointestinal tract, even during processing. The same applies to the pH and temperature fluctuations of nutritional compositions that have been exposed to typical pasteurization-like processing conditions. Examples of such pathogens include enterotoxic E. coli (ETEC), enteropathogenic E. coli (EPEC), Haemophilus influenza , Shiga toxin-producing E. coli (STEC), enteroaggregative E. coli ( EAEC), Salmonella ser. Typhimurium, Shigella flexneri , Rotavirus, Norovirus, Respiratory Fusion Virus (RSV), Adenovirus and combinations thereof.
本文提供新穎的營養產品,其容易消化、提供生理化學效益和/或提供生理效益。在本文之一實施例中,營養組成物包含脂質或脂肪、蛋白質來源、具有至少20%寡醣的益菌生組成物,特別為包含半乳糖寡醣(GOS)者、以及乳鐵蛋白,其提供對抗存於人胃腸道中不希望的細菌菌株的活性抗入侵或抗附著機轉。包含在該組成物中之該乳鐵蛋白係藉由非人來源所製造。在某些實施例中,該益菌生包含半乳糖寡醣及聚葡萄糖之組合。更特定言之,本文中所揭露之組成物包含:a.至高達約7 g/100 kcal的脂肪或脂質,更佳為約3至約7 g/100 kcal的脂肪或脂質;b.至高達約5 g/100 kcal的蛋白質來源,更佳為約1至約5 g/100 kcal的蛋白質來源;c.約0.1 g/100 kcal至約1 g/100 kcal之益菌生組成物,其包含聚葡萄糖和/或半乳糖寡醣。更佳的是,該營養組成物包含約0.3 g/100 kcal至約0.7 g/100 kcal的益菌生組成物,其包含聚葡萄糖及半乳糖寡醣之組合;以及 d.至少約10 mg/100kCal的乳鐵蛋白,更佳為約70 mg至約220 mg/100kCal的乳鐵蛋白,且最佳為約90 mg至約190 mg/100kCal的乳鐵蛋白。 Provided herein are novel nutritional products that are easy to digest, provide physiochemical benefits, and / or provide physiological benefits. In one embodiment herein, the nutritional composition comprises a lipid or fat, a protein source, a probiotic composition with at least 20% oligosaccharides, particularly a galactose oligosaccharide (GOS), and lactoferrin, Provides active anti-invasion or anti-adhesion machinery against unwanted bacterial strains present in the human gastrointestinal tract. The lactoferrin contained in the composition is manufactured from a non-human source. In certain embodiments, the probiotic contains a combination of galactooligosaccharide and polydextrose. More specifically, the composition disclosed herein comprises: a. Up to about 7 g / 100 kcal of fat or lipid, more preferably about 3 to about 7 g / 100 kcal of fat or lipid; b. Up to A protein source of about 5 g / 100 kcal, more preferably a protein source of about 1 to about 5 g / 100 kcal; c. A probiotic composition of about 0.1 g / 100 kcal to about 1 g / 100 kcal, comprising Polydextrose and / or galactooligosaccharide. More preferably, the nutritional composition comprises a probiotic composition of about 0.3 g / 100 kcal to about 0.7 g / 100 kcal, which comprises a combination of polydextrose and galactooligosaccharides; and d. Lactoferrin of at least about 10 mg / 100 kCal, more preferably about 70 mg to about 220 mg / 100 kCal of lactoferrin, and most preferably about 90 mg to about 190 mg / 100 kCal of lactoferrin.
如本文中所用,術語「益菌生」意指藉由選擇性刺激結腸(colon)中可改善宿主健康的一種或少數種細菌生長和/或活性來正面影響宿主的非可消化性食物成分。 As used herein, the term "probiotics" means non-digestible food ingredients that positively affect a host by selectively stimulating the growth and / or activity of one or a few bacteria in the colon that can improve host health.
術語「益生菌」意指對宿主健康具有正面作用的低致病性或不具致病性微生物。 The term "probiotic" means a low-pathogenic or non-pathogenic microorganism that has a positive effect on the health of the host.
如本文中所用,術語「嬰兒」通常係定義為約出生至12個月大的人。 As used herein, the term "infant" is generally defined as a person from about birth to 12 months of age.
「早產嬰兒」為妊娠少於約37週後出生的嬰兒。 "Premature babies" are babies born after about 37 weeks of pregnancy.
如本文中所用,「足月嬰兒」意指妊娠至少37週後出生之嬰兒。 As used herein, "term infant" means an infant born after at least 37 weeks of pregnancy.
「孩童」係定義為年齡從12個月至約12歲的人。 "Child" is defined as someone from 12 months to about 12 years old.
如本文中所用,「抗細菌有效量」通常定義為提供至少一種對抗細菌菌株之抗入侵機轉的乳鐵蛋白量。 As used herein, an "antibacterially effective amount" is generally defined as the amount of lactoferrin that provides at least one anti-invasion mechanism against a bacterial strain.
如本文中所用,「由非人來源生產之乳鐵蛋白」意指藉由人乳以外的來源製造或獲得之乳鐵蛋白。舉例而言,用於本文之乳鐵蛋白包括藉由經基因修飾之生物體所製造的人類乳鐵蛋白和非人乳鐵蛋白。 As used herein, "lactoferrin produced from non-human sources" means lactoferrin manufactured or obtained from sources other than human milk. For example, lactoferrin as used herein includes human lactoferrin and non-human lactoferrin produced by genetically modified organisms.
「生物活性乳鐵蛋白」意指具有至少一種對抗病原菌之抗入侵或抗附著機轉的乳鐵蛋白。 "Bioactive lactoferrin" means lactoferrin with at least one anti-invasion or anti-adhesion mechanism against pathogenic bacteria.
如本文中所用,術語「非人乳鐵蛋白」指具有與人乳鐵蛋白胺基酸序列不同的胺基酸序列之乳鐵蛋白。 As used herein, the term "non-human lactoferrin" refers to lactoferrin having an amino acid sequence that is different from the human lactoferrin amino acid sequence.
如本文中所用,術語「生物體(organism)」指任何持續之生命系統,例如動物、植物、真菌或微生物。 As used herein, the term "organism" refers to any continuous living system, such as an animal, plant, fungus, or microorganism.
如本文中所用,術語「模擬」意指具有或採取某形式或外觀的,或具有或產生症狀類似的。 As used herein, the term "mimic" means having or taking some form or appearance, or having or producing similar symptoms.
在一些實施例中,該營養產品可為嬰兒配方。術語「嬰兒配方」係指為人乳取代物,其呈液體或粉末形式、能滿足嬰兒營養分需求的組成物。在美國,嬰兒配方含量規定於美國聯邦法規第21章第100、106及107部份。這些法規界定大量營養素、維生素、礦物質及其他成分量以模擬人乳的營養及其他性質。在另一實施例中,該營養產品可為人乳強化營養品,意指其為加至人乳中以加強人乳營養價值之組成物。作為人乳強化營養品,該揭露之組成物可為粉末形式或液體形式。在又另一實施例中,本文之營養產品可為孩童的營養組成物。 In some embodiments, the nutritional product may be an infant formula. The term "infant formula" refers to a composition that is a substitute for human milk that is in liquid or powder form and meets the nutritional needs of infants. In the United States, infant formula levels are specified in Title 21, Sections 100, 106, and 107 of the Code of Federal Regulations. These regulations define large amounts of nutrients, vitamins, minerals and other ingredients to mimic the nutritional and other properties of human milk. In another embodiment, the nutritional product may be a human milk fortified product, which means that it is a composition added to human milk to enhance the nutritional value of human milk. As a human milk fortified nutritional product, the disclosed composition may be in powder form or liquid form. In yet another embodiment, the nutritional product herein may be a nutritional composition for children.
本文之營養產品可提供最低、部分或完全營養支持。該組成物可為營養補充品或代餐。在一些實施例中,該組成物可連同食物或營養組成物而被投予。在此實施例中,該組成物可於個體攝取前與食物或其他營養組成物互混,或可於個體攝取食物或營養組成物之前或之後投予至該個體。該組成物可投予至接受嬰兒配方、母乳、人乳強化營 養品或彼等之組合的早產嬰兒。 The nutritional products in this article can provide minimal, partial, or full nutritional support. The composition may be a nutritional supplement or a meal replacement. In some embodiments, the composition may be administered along with a food or nutritional composition. In this embodiment, the composition may be intermixed with food or other nutritional composition prior to ingestion by the individual, or may be administered to the individual before or after ingestion of the food or nutritional composition by the individual. This composition can be administered to infant formula, breast milk, and human milk fortification camps. Premature babies with nutrition or their combination.
該組成物可為(但不必要為)營養完整。熟習本技術人士將理解「營養完整」取決於許多因素而有不同,包括但不限於該術語所適用之個體年齡、臨床條件及膳食攝入。一般而言,「營養完整」意指本文之營養組成物提供所有正常生長所需的適量之碳水化合物、脂質、必需脂肪酸、蛋白質、必需胺基酸、條件性必需胺基酸、維生素、礦物質及能量。若在指涉營養素,術語「必需」係指任何身體無法合成足以供正常生長以及維持健康而因此必須藉由膳食供應的營養素。術語「條件性必需」若在指涉營養素,則意指該營養素在當身體無法取得適量前驅化合物以供進行內生性合成的條件下必須藉由膳食供應。 The composition may be (but not necessarily) nutritionally complete. Those skilled in the art will understand that "nutrition integrity" varies depending on many factors, including but not limited to the age of the individual to which the term applies, clinical conditions, and dietary intake. In general, "nutritive integrity" means that the nutritional composition herein provides all the appropriate amounts of carbohydrates, lipids, essential fatty acids, proteins, essential amino acids, conditional essential amino acids, vitamins, minerals required for normal growth. And energy. When referring to nutrients, the term "essential" refers to any nutrient that the body cannot synthesize enough for normal growth and maintenance of health and therefore must be supplied through the diet. The term "conditionally necessary" when referring to a nutrient means that the nutrient must be supplied through the diet under conditions where the body cannot obtain a suitable amount of precursor compounds for endogenous synthesis.
針對早產嬰兒,該「營養完整」之組成物依定義為提供該早產嬰兒所有生長所需的質量上適量之碳水化合物、脂質、必需脂肪酸、蛋白質、必需胺基酸、條件性必需胺基酸、維生素、礦物質及能量。針對足月嬰兒,該「營養完整」之組成物依定義為提供該足月嬰兒所有生長所需的質量上適量之碳水化合物、脂質、必需脂肪酸、蛋白質、必需胺基酸、條件性必需胺基酸、維生素、礦物質及能量。針對孩童,該「營養完整」之組成物依定義為提供該孩童所有生長所需的質量上適量之碳水化合物、脂質、必需脂肪酸、蛋白質、必需胺基酸、條件性必需胺基酸、維生素、礦物質及能量。 For premature babies, the "nutritional integrity" composition is defined as providing a suitable amount of carbohydrates, lipids, essential fatty acids, proteins, essential amino acids, conditional essential amino acids, Vitamins, minerals and energy. For term infants, the "nutritional integrity" composition is defined as providing the appropriate amount of carbohydrates, lipids, essential fatty acids, proteins, essential amino acids, and conditional essential amino groups required for all growth of the term infant Acids, vitamins, minerals and energy. For children, the "nutritive integrity" composition is defined as providing the child with all the quality carbohydrates, lipids, essential fatty acids, proteins, essential amino acids, conditional essential amino acids, vitamins, Minerals and energy.
該營養組成物可用任何本技術領域中已知形式提供, 包括粉末、凝膠、懸浮液、糊劑、固體、液體、液體濃縮物或現成品。在一較佳實施例中,該營養組成物係嬰兒配方,特別適於用作為嬰兒唯一來源營養的嬰兒配方。 The nutritional composition may be provided in any form known in the art, These include powders, gels, suspensions, pastes, solids, liquids, liquid concentrates or ready-made products. In a preferred embodiment, the nutritional composition is an infant formula, which is particularly suitable for use as an infant formula as the sole source of nutrition for infants.
在較佳實施例中,本文所揭露之營養產品可經腸道投予。如本文所用,「腸道」意指透過胃腸道或消化道或於胃腸道或消化道內,且「經腸道投予」包括口服餵食、胃內餵食、鼻腸管投服(transpyloric administration)或任何其他導入消化道。 In a preferred embodiment, the nutritional products disclosed herein can be administered enterally. As used herein, "intestinal tract" means through or within the gastrointestinal or digestive tract, and "parenteral administration" includes oral feeding, intragastric feeding, transpyloric administration, or Any other lead into the digestive tract.
乳鐵蛋白為約80 kD的單鏈多肽,取決於物種而含有1-4個聚醣。不同物種的乳鐵蛋白3-D結構均非常相似,但並非完全相同。各乳鐵蛋白包含二個同型葉狀結構,稱之為N-葉及C-葉,係分別指分子N端及C端。各葉狀結構進一步由二個次葉或結構域組成,其形成裂隙,其中鐵離子(Fe3+)係與(二)碳酸鹽陰離子協同合作而緊密結合。這些結構域係分別稱為N1、N2、C1及C2。乳鐵蛋白的N端具有強陽離子胜肽區域,其負責許多重要結合特性。乳鐵蛋白具有非常高的等電點(~pI 9)且其陽離子本質在其防禦細菌、病毒及真菌病原體的能力中扮演主要角色。於乳鐵蛋白N端區域內有數群陽離子胺基酸殘基調介乳鐵蛋白抗各類微生物的生物活性。舉例而言,人類乳鐵蛋白的N端殘基1-47(牛乳鐵蛋白為1-48)對乳鐵蛋白的鐵非依賴性生物活性係為關鍵。在人類乳鐵蛋白中,在N端中的殘基2至5(RRRR)及28至31(RKVR)為富含精胺酸陽離子結構域,對乳鐵蛋白的抗微生物活性特別 重要。在牛乳鐵蛋白中亦發現相似N端區域(殘基17至42;FKCRRWQWRMKKLGAPSITCVRRAFA)。 Lactoferrin is a single-chain polypeptide of about 80 kD and contains 1-4 glycans depending on the species. The 3-D structures of lactoferrin in different species are very similar, but not exactly the same. Each lactoferrin contains two homotypic leaf-like structures, called N-lobe and C-lobe, which refer to the N- and C-terminus of the molecule, respectively. Each leaf-like structure is further composed of two secondary leaves or domains, which form a fissure, in which the iron ion (Fe 3+ ) and the (di) carbonate anion cooperate in close cooperation. These domain systems are called N1, N2, C1, and C2, respectively. The N-terminus of lactoferrin has a strong cationic peptide region, which is responsible for many important binding properties. Lactoferrin has a very high isoelectric point (~ pI 9) and its cationic nature plays a major role in its ability to defend against bacterial, viral and fungal pathogens. There are several groups of cationic amino acid residues in the N-terminal region of lactoferrin to mediate the biological activity of lactoferrin against various microorganisms. For example, N-terminal residues 1-47 of human lactoferrin (1-48 for bovine lactoferrin) are critical to the iron-independent biological activity of lactoferrin. In human lactoferrin, residues 2 to 5 (RRRR) and 28 to 31 (RKVR) at the N-terminus are rich in arginine cation domains, which are particularly important for the antimicrobial activity of lactoferrin. A similar N-terminal region (residues 17 to 42; FKCRRWQWRMKKLGAPSITCVRRAFA) was also found in bovine lactoferrin.
如在「Perspectives on Interactions Between Lactoferrin and Bacteria」中所述(該文出現在BIOCHEMISTRY AND CELL BIOLOGY,pp 275-281(2006)刊物中),來自不同宿主物種的乳鐵蛋白雖然通常具有相對高等電點,在內葉末端區域帶有正電胺基酸,但胺基酸序列不同。適合用於本文之乳鐵蛋白包括具有與HLf(349-364)片段的胺基酸序列AVGEQELRKCNQWSGL至少48%同源性。在一些實施例中,該乳鐵蛋白具有與HLf(349-364)片段的胺基酸序列AVGEQELRKCNQWSGL至少65%同源性,且在實施例中,為至少75%同源性。舉例而言,用於本文之非人乳鐵蛋白包括而不限於牛乳鐵蛋白、豬乳鐵蛋白、馬乳鐵蛋白、水牛乳鐵蛋白、山羊乳鐵蛋白、鼠類乳鐵蛋白及駱駝乳鐵蛋白。 As described in Perspectives on Interactions Between Lactoferrin and Bacteria (this article appears in BIOCHEMISTRY AND CELL BIOLOGY, pp. 275-281 (2006)), lactoferrin from different host species although usually has a relatively high isoelectric point , With positively charged amino acids in the inner lobe terminal region, but the amino acid sequence is different. Suitable lactoferrin for use herein includes having at least 48% homology to the amino acid sequence AVGEQELRKCNQWSGL of the HLf (349-364) fragment. In some embodiments, the lactoferrin has at least 65% homology to the amino acid sequence AVGEQELRKCNQWSGL of the HLf (349-364) fragment, and in embodiments, at least 75% homology. For example, non-human lactoferrin as used herein includes, but is not limited to, bovine lactoferrin, porcine lactoferrin, horse lactoferrin, buffalo lactoferrin, goat lactoferrin, murine lactoferrin, and camel lactoferrin protein.
令人驚訝的是,此處所包含之乳鐵蛋白形式即使暴露於低pH(即低於7,且甚至低至約4.6或更低)和/或高溫(即超過約65℃,且高至約120℃)、為預期會破壞或嚴重地限制人乳鐵蛋白或重組人乳鐵蛋白穩定性或活性之條下,仍維持相關活性。用於本文所述之營養組成物種類的某些加工方式過程中,可預期這些低pH和/或高溫條件,例如巴斯德殺菌法。舉例而言,在牛乳鐵蛋白的胺基酸組成僅具有與人乳鐵蛋白約70%序列同源性、且在人或重組人乳鐵蛋白變得不穩定或無活性的條件下係穩定及仍具 活性的同時,牛乳鐵蛋白具有對抗人腸道中所發現之非所欲細菌病原的殺菌活性。 Surprisingly, the lactoferrin forms contained herein are even exposed to low pH (ie, less than 7, and even as low as about 4.6 or less) and / or high temperatures (ie, more than about 65 ° C, and as high as about 120 ° C), which is expected to disrupt or severely limit the stability or activity of human lactoferrin or recombinant human lactoferrin, and still maintain the relevant activity. These low pH and / or high temperature conditions may be expected during certain processing methods for the types of nutritional compositions described herein, such as pasteurization. For example, the amino acid composition of bovine lactoferrin has only about 70% sequence homology with human lactoferrin, and is stable under conditions where human or recombinant human lactoferrin becomes unstable or inactive, and Still with At the same time, bovine lactoferrin has a fungicidal activity against undesired bacterial pathogens found in the human intestine.
在美國專利第4,791,193號(其係以引用方式全部併入本案)中,Okonogi等人揭露用於製造高純度牛乳鐵蛋白的方法。通常,該方法如揭露者,包括三個步驟。首先生乳原料與弱酸性陽離子交換劑接觸以吸附乳鐵蛋白,接以第二步驟,在此步驟進行沖洗以移除未吸附物質。接著為去吸附步驟,此時乳鐵蛋白被移走,以產生經純化之牛乳鐵蛋白。其他方法可包括如美國專利第7,368,141、5,849,885、5,919,913及5,861,491號中所述步驟,該等揭露均以引用方式全部併入本案。 In U.S. Patent No. 4,791,193, which is incorporated by reference in its entirety, Okonogi et al. Disclose a method for manufacturing high-purity bovine lactoferrin. Generally, the method, as disclosed, involves three steps. The raw milk raw material is first contacted with a weakly acidic cation exchanger to adsorb lactoferrin, followed by a second step, in which a rinse is performed to remove unadsorbed material. This is followed by a desorption step, at which time the lactoferrin is removed to produce purified bovine lactoferrin. Other methods may include steps as described in US Patent Nos. 7,368,141, 5,849,885, 5,919,913, and 5,861,491, all of which are incorporated herein by reference in their entirety.
在一實施例中,乳鐵蛋白存在於該營養組成物中的量為至少約10 mg/100 kCal,特別是當希望該營養組成物係欲供孩童之用。在某些實施例中,乳鐵蛋白之上限係約240 mg/100 kCal。在另一實施例中,其中該營養組成物係嬰兒配方,乳鐵蛋白存在於該營養組成物中的量為約70 mg至約220 mg/100 kCal;在又另一實施例中,乳鐵蛋白存在的量為約90 mg至約190 mg/100 kCal。針對嬰兒的營養組成物可包括乳鐵蛋白量為每升配方約0.5 mg至約1.5 mg。在取代人乳的營養組成物,乳鐵蛋白含量可為每升配方約0.6 mg至約1.3 mg。 In one embodiment, lactoferrin is present in the nutritional composition in an amount of at least about 10 mg / 100 kCal, especially when the nutritional composition is desired for children. In some embodiments, the upper limit for lactoferrin is about 240 mg / 100 kCal. In another embodiment, wherein the nutritional composition is an infant formula, lactoferrin is present in the nutritional composition in an amount of about 70 mg to about 220 mg / 100 kCal; in still another embodiment, lactoferrin The protein is present in an amount of about 90 mg to about 190 mg / 100 kCal. Nutritional compositions for infants may include lactoferrin in an amount of about 0.5 mg to about 1.5 mg per liter of formula. In replacing the nutritional composition of human milk, the content of lactoferrin may be from about 0.6 mg to about 1.3 mg per liter of the formula.
實施本發明的適合之脂肪或脂質來源可為任何本技術領域中已知或使用者,包括但不限於動物來源,例如:乳脂(milk fat)、乳酪(butter)、乳酪脂(butter fat)、 蛋黃脂質;水產來源,例如:魚油、水產油脂、單細胞油;蔬菜及植物油脂,例如:玉米油、菜籽油、葵花子油、大豆油、軟棕櫚油(palmolein)、椰子油、高油酸葵花子油、月見草油、芥花子油、橄欖油、亞麻子油、棉籽油、高油酸紅花子油、硬棕櫚油(palm stearin)、棕櫚仁油、小麥胚芽油;中鏈三酸甘油脂及脂肪酸之乳化劑及酯類;及任何彼等之組合。 A suitable source of fat or lipid for practicing the present invention may be any known or user in the art, including but not limited to animal sources, such as: milk fat, butter, butter fat, Egg yolk lipids; aquatic sources such as: fish oil, aquatic fats, single-cell oils; vegetable and vegetable fats such as corn oil, rapeseed oil, sunflower oil, soybean oil, palmolein, coconut oil, high oleic acid Sunflower oil, evening primrose oil, canola oil, olive oil, linseed oil, cottonseed oil, high oleic safflower oil, palm stearin, palm kernel oil, wheat germ oil; medium chain triglycerides and Emulsifiers and esters of fatty acids; and any combination thereof.
可用於實施本文的牛乳蛋白來源包括但不限於乳蛋白粉末、乳蛋白濃縮物、乳蛋白分離物、脫脂奶固體、脫脂奶、脫脂乳粉、乳清蛋白、乳清蛋白分離物、乳清蛋白濃縮物、甜乳清、酸乳清、酪蛋白、酸酪蛋白、酪蛋白鹽(例如酪蛋白鈉、酪蛋白鈉鈣、酪蛋白鈣)及任何彼等之組合。 Sources of cow's milk proteins useful in the practice herein include, but are not limited to, milk protein powder, milk protein concentrate, milk protein isolate, skim milk solids, skim milk, skim milk powder, whey protein, whey protein isolate, whey protein Concentrates, sweet whey, sour whey, casein, sour casein, casein salts (e.g. sodium caseinate, sodium caseinate calcium, caseinate calcium) and any combination thereof.
在一實施例中,該蛋白質係以完整蛋白質提供。在其他實施例中,該蛋白質係以完整蛋白質及部分水解蛋白質兩者來提供,水解程度介於約4%至10%。在又另一實施例中,該蛋白質來源可以含麩醯胺酸胜肽補充。 In one embodiment, the protein is provided as a whole protein. In other embodiments, the protein is provided as both a whole protein and a partially hydrolyzed protein, with a degree of hydrolysis ranging from about 4% to 10%. In yet another embodiment, the protein source may be supplemented with a glutamate-containing peptide.
在本文之一特定實施例中,該蛋白質來源之乳清:酪蛋白比例係類似於人乳中之比例。在一實施例中,該蛋白質來源包含約20%至約85%乳清蛋白。在另一實施例中,該蛋白質來源可包含約20%至約80%酪蛋白。在本文之又另一實施例中,該蛋白質來源包括約60%至約85%乳清以及約15%至約40%酪蛋白。 In a specific embodiment herein, the protein-derived whey: casein ratio is similar to that in human milk. In one embodiment, the protein source comprises about 20% to about 85% whey protein. In another embodiment, the protein source may include about 20% to about 80% casein. In yet another embodiment herein, the protein source includes about 60% to about 85% whey and about 15% to about 40% casein.
在本文之一實施例中,該營養組成物亦可含有一或多 種益生菌。只要能達到預期結果,在此實施例中,任何本技術領域已知的益生菌均可接受。在一特定實施例中,該益生菌可選自乳酸桿菌(Lactobacillus)菌種、鼠李醣乳酸桿菌(Lactobacillus rhamnosus GG)、雙叉桿菌(Bifidobacterium)菌種、龍根雙叉桿菌(Bifidobacterium longum)、短雙叉桿菌(Bifidobacterium brevis)及動物雙叉桿菌(Bifidobacterium animalis subsp.lactis BB-12)。 In one embodiment herein, the nutritional composition may also contain one or more probiotics. As long as the desired results are achieved, any probiotics known in the art are acceptable in this example. In a specific embodiment, the probiotic bacteria may be selected from Lactobacillus strains, Lactobacillus rhamnosus GG, Bifidobacterium strains, Bifidobacterium longum strains. , Bifidobacterium brevis and Bifidobacterium animalis subsp . Lactis BB-12.
若包括在該組成物中,則該益生菌量可為每日每kg體重約104至約1010菌落形成單位(cfu)。在另一實施例中,該益生菌量可為每日每kg體重約106至約109cfu。在又另一實施例中,該益生菌量可為每日每kg體重至少約106 cfu。又,該揭露之組成物亦可包括益生菌態(probiotic-conditioned)介質組分。 If included in the composition, the amount of the probiotic may be formed units (cfu) per day is from about 104 to about 1010 colonies per kg body weight. In another embodiment, the amount of probiotics may be about 10 6 to about 10 9 cfu per kg of body weight per day. In yet another embodiment, the amount of probiotics may be at least about 10 6 cfu per kg of body weight per day. In addition, the disclosed composition may also include a probiotic-conditioned medium component.
在一實施例中,該益生菌可為活菌或非活菌。如本文中所用,術語「活菌」係指活的微生物。術語「非活菌」或「非活益生菌」意指非活的益生菌微生物、它們的細胞組成和其代謝物。此等非活益生菌可為經過熱殺死或其他不活化、但仍留存對宿主健康有利影響能力。在本文中,可用的益生菌可為天然存在、合成或透過遺傳操作生物體所開發,而不論此種新來源為現今已知或之後所開發者。 In one embodiment, the probiotic bacteria may be live bacteria or non-live bacteria. As used herein, the term "live bacteria" refers to living microorganisms. The term "non-living bacteria" or "non-living probiotics" means non-living probiotic microorganisms, their cellular composition and their metabolites. These non-living probiotic bacteria can be heat-killed or otherwise inactivated, but still retain the ability to have a beneficial effect on host health. In this context, the probiotics available may be developed by naturally occurring, synthetic or genetically manipulated organisms, whether such new sources are now known or later developed.
營養組成物含有一或多種益菌生。此種益菌生可為天然存在、合成或透過遺傳操作生物體和/或植物所開發,而不論此種新來源為現今已知或之後所開發者。在某些實 施例中,包括於本文的組成物之益菌生包括美國專利第7,572,474號所教示,該案之揭露係以引用方式併入本案。 The nutritional composition contains one or more probiotics. Such probiotics can be developed by naturally occurring, synthetic or genetically manipulated organisms and / or plants, regardless of whether the new source is now known or later developed. In some real In the examples, the probiotics of the compositions included herein include the teachings of US Patent No. 7,572,474, the disclosure of which is incorporated herein by reference.
可用於本文之益菌生可包括寡醣、多醣及其他含有果糖、木糖、大豆、半乳糖、葡萄糖及甘露糖之益菌生。更具體而言,有用於本文之益菌生可包括乳果糖、乳果寡醣(lactosucrose)、棉子糖、葡萄糖寡醣、菊糖、聚葡萄糖、聚葡萄糖粉末、半乳糖寡醣、果糖寡醣、異麥芽糖寡醣、大豆寡醣、乳果寡醣、木糖寡醣、幾丁寡醣(chito-oligosaccharide)、甘露糖寡醣、阿拉伯糖寡醣、唾液酸寡醣、海藻糖寡醣、及龍膽寡醣(gentio-oligosaccharide)。 Probiotics useful herein may include oligosaccharides, polysaccharides, and other probiotics containing fructose, xylose, soy, galactose, glucose, and mannose. More specifically, the probiotics useful herein may include lactulose, lactosucrose, raffinose, gluco-oligosaccharides, inulin, polydextrose, polydextrose powder, galactooligosaccharides, and fructo-oligosaccharides. Sugar, isomalt oligosaccharide, soy oligosaccharide, lactulose oligosaccharide, xylose oligosaccharide, chito-oligosaccharide, mannose oligosaccharide, arabinose oligosaccharide, sialic acid oligosaccharide, trehalose oligosaccharide , And gentio-oligosaccharide.
在一實施例中,在該營養組成物中所存在的益菌生總量於每公升該組成物可占約1.0 g至約10.0 g。或者,在該營養組成物中所存在的益菌生總量於每公升該組成物可占約2.0 g至約8.0 g。該益菌生的至少20%應包含半乳糖寡醣(GOS)、聚葡萄糖或其混合物。較佳的是,該營養組成物包含聚葡萄糖和半乳糖寡醣。任意地,除了聚葡萄糖和/或半乳糖寡醣,該營養組成物包含一或多種額外益菌生。在一實施例中,該營養組成物中的半乳糖寡醣和/或聚葡萄糖各含量範圍可為約1.0 g/L至約4.0 g/L。 In one embodiment, the total amount of probiotics present in the nutritional composition may be about 1.0 g to about 10.0 g per liter of the composition. Alternatively, the total amount of probiotics present in the nutritional composition may be about 2.0 g to about 8.0 g per liter of the composition. At least 20% of the probiotics should contain galactose oligosaccharides (GOS), polydextrose, or mixtures thereof. Preferably, the nutritional composition comprises polydextrose and galactooligosaccharide. Optionally, in addition to polydextrose and / or galactooligosaccharide, the nutritional composition comprises one or more additional probiotics. In one embodiment, each content of galactooligosaccharide and / or polyglucose in the nutritional composition may range from about 1.0 g / L to about 4.0 g / L.
在一實施例中,存在於該營養組成物中的益菌生總量可為約0.1 g/100 kcal至約1 g/100 kcal。更佳的是,存於該營養組成物中的益菌生總量可約0.3 g/100 kcal至約0.7 g/100 kcal。該益菌生至少20%應包含半乳糖寡醣(GOS)和/或聚葡萄糖(PDX)。 In one embodiment, the total amount of probiotics present in the nutritional composition may be about 0.1 g / 100 kcal to about 1 g / 100 kcal. More preferably, the total amount of probiotics stored in the nutritional composition may be about 0.3 g / 100 kcal to about 0.7 g / 100 kcal. At least 20% of this probiotic should contain galactose oligosaccharides (GOS) and / or polydextrose (PDX).
在一實施例中,該營養組成物中之半乳糖寡醣量可為約0.2 g/100 Kcal至約1.0 g/100 Kcal。在另一實施例中,該營養組成物中的半乳糖寡醣量可為約0.1 g/100 Kcal至約0.5 g/100 Kcal。在另一實施例中,半乳糖寡醣量係約0.2 g/100 kcal至約0.6 g/100 kcal的範圍內。在一實施例中,若使用聚葡萄糖作為益菌生,則該營養組成物中的聚葡萄糖量範圍可為約0.1 g/100 kcal至約1 g/100 kcal。在另一實施例中,聚葡萄糖量係約0.2 g/100 kcal至約0.6 g/100 kcal的範圍內。在又另一實施例中,若在該益菌生組成物中使用聚葡萄糖,則在一實施例中,該營養組成物中的聚葡萄糖量範圍可為約0.1 g/100 Kcal至約0.5 g/100 Kcal的範圍內。在某些實施例中,該益菌生組成物中的聚葡萄糖與半乳糖寡醣比例係介於約9:1至約1:9。 In one embodiment, the amount of galactooligosaccharide in the nutritional composition may be about 0.2 g / 100 Kcal to about 1.0 g / 100 Kcal. In another embodiment, the amount of galactooligosaccharide in the nutritional composition may be about 0.1 g / 100 Kcal to about 0.5 g / 100 Kcal. In another embodiment, the amount of galactooligosaccharide is in the range of about 0.2 g / 100 kcal to about 0.6 g / 100 kcal. In one embodiment, if polydextrose is used as the probiotic, the amount of polydextrose in the nutritional composition may range from about 0.1 g / 100 kcal to about 1 g / 100 kcal. In another embodiment, the amount of polydextrose ranges from about 0.2 g / 100 kcal to about 0.6 g / 100 kcal. In yet another embodiment, if polydextrose is used in the probiotic composition, in one embodiment, the amount of polydextrose in the nutritional composition may range from about 0.1 g / 100 Kcal to about 0.5 g. / 100 Kcal. In certain embodiments, the ratio of polydextrose to galactooligosaccharide in the probiotic composition is between about 9: 1 to about 1: 9.
較佳的是,當提供該營養組成物予人時,該益菌生組成物合併乳鐵蛋白抑制胃腸道中一或多種病原體的附著。此種病原體之一實例係阪崎腸桿菌(Enterobacter sakazakii)(或又名阪崎克隆納斯桿菌(Cronobacter sakazakii))。另一病原體為大腸桿菌,其附著係藉由合併乳鐵蛋白與該益菌生組成物所抑制。 Preferably, when the nutritional composition is provided to a human, the probiotic composition combined with lactoferrin inhibits the adhesion of one or more pathogens in the gastrointestinal tract. One example of such a pathogen is Enterobacter sakazakii (or aka Cronobacter sakazakii ). Another pathogen is E. coli, whose attachment is inhibited by combining lactoferrin with the probiotic composition.
本文之營養配方亦可含有長鏈多不飽和脂肪酸(LCPUFA)來源,其包含二十二碳六烯酸(DHA)。其他適合的LCPUFA包括但不限於α-亞麻油酸、γ-亞麻油酸、 亞麻油酸、次亞麻油酸、二十碳五烯酸(EPA)及花生四烯酸(ARA)。 The nutritional formulas herein may also contain a long-chain polyunsaturated fatty acid (LCPUFA) source, which contains docosahexaenoic acid (DHA). Other suitable LCPUFAs include, but are not limited to, alpha-linolenic acid, gamma-linolenic acid, Linoleic acid, hypolinoleic acid, eicosapentaenoic acid (EPA) and arachidonic acid (ARA).
在一實施例中,該營養組成物係以DHA及ARA兩者補充。在此實施例中,ARA:DHA重量比例可為約1:3至約9:1。在本文的一實施例中,此比例係約1:2至約4:1。 In one embodiment, the nutritional composition is supplemented with both DHA and ARA. In this embodiment, the ARA: DHA weight ratio may be from about 1: 3 to about 9: 1. In an embodiment herein, the ratio is about 1: 2 to about 4: 1.
該營養組成物中的長鏈多不飽和脂肪酸量可為約5 mg/100 kcal至約100 mg/100 kcal,更佳為約10 mg/100 kcal至約50 mg/100 kcal。 The amount of long-chain polyunsaturated fatty acids in the nutritional composition may be about 5 mg / 100 kcal to about 100 mg / 100 kcal, and more preferably about 10 mg / 100 kcal to about 50 mg / 100 kcal.
該營養組成物可使用本技術領域中已知的標準技術補充含有DHA及ARA的油脂。舉例而言,DHA及ARA可藉由取代等量之通常存在於配方中的油脂(例如高油酸葵花子油)來加至該配方。作為另一實例,含有DHA及ARA的油脂可藉由取代等量之通常存在於未有DHA及ARA配方中的其餘整體脂肪摻合物來加至該配方。 The nutritional composition can be supplemented with fats containing DHA and ARA using standard techniques known in the art. For example, DHA and ARA can be added to the formula by replacing equivalent amounts of fats and oils typically present in the formula (such as high oleic sunflower oil). As another example, fats and oils containing DHA and ARA can be added to the formulation by replacing equivalent amounts of the remaining whole fat blends that are normally present in formulations without DHA and ARA.
若有使用,則DHA及ARA來源可為本技術領域中任何已知來源,例如水產油脂、魚油、單細胞油脂、蛋黃脂質及腦部脂質。在一些實施例中,該DHA及ARA係分別源自單細胞Martek油脂、DHASCO®及ARASCO®或彼等之變化物。該DHA及ARA可為天然形式,惟該LCPUFA來源之剩餘物不會造成對嬰兒任何實質有害影響。或者,該DHA及ARA可以精製形式使用。 If used, DHA and ARA sources can be any known source in the art, such as aquatic oils, fish oils, single-cell oils, egg yolk lipids, and brain lipids. In some embodiments, the DHA and ARA are derived from single-cell Martek oil, DHASCO® and ARASCO®, or variants thereof. The DHA and ARA can be in natural form, but the residue of the LCPUFA source will not cause any substantial harmful effects on the baby. Alternatively, the DHA and ARA can be used in a refined form.
在本文的一實施例中,DHA及ARA來源係如美國專利第5,374,567、5,550,156及5,397,591號中所教示之單細胞油脂,該等專利所揭露者係以引用方式全部併入本案 。然而,本文不只限於此等油脂。 In an embodiment herein, the sources of DHA and ARA are single-cell lipids as taught in U.S. Pat. Nos. 5,374,567, 5,550,156, and 5,397,591. The disclosures of these patents are incorporated herein by reference in their entirety. . However, this article is not limited to these greases.
在某些實施例中,該營養組成物包含約0.5 mg/100 kcal至約5 mg/100 kcal的鐵,包括與乳鐵蛋白結合之鐵。 In certain embodiments, the nutritional composition comprises about 0.5 mg / 100 kcal to about 5 mg / 100 kcal of iron, including iron bound to lactoferrin.
如用於本文之實施例的乳鐵蛋白之效益係其在人胃腸道中之抗入侵及抗附著機轉。尤其,乳鐵蛋白可破壞某些細菌入侵並造成發病所使用之注射針。同樣地,乳鐵蛋白抑制病原體在人胃腸道中的附著。已知造成發病的此類細菌之一實例為大腸桿菌,其可造成嬰兒、孩童及成人腹瀉,且被認為是小兒腹瀉的起因。大腸桿菌製造細菌蛋白並透過針狀複體(needle complex)經由第三型分泌系統移位細菌蛋白。 The benefits of lactoferrin as used in the examples herein are its resistance to invasion and resistance to attachment in the human gastrointestinal tract. In particular, lactoferrin can destroy certain bacterial invasions and cause injection needles used for pathogenesis. Similarly, lactoferrin inhibits the adhesion of pathogens in the human gastrointestinal tract. One example of such bacteria known to cause disease is E. coli, which can cause diarrhea in infants, children, and adults, and is considered to be the cause of diarrhea in children. E. coli makes bacterial proteins and translocates them through a needle complex via a type III secretion system.
許多革蘭氏陰性致病菌的分泌系統為第三型分泌系統,其中包括的細菌如下:志賀桿菌屬、沙門桿菌屬、假單孢菌屬及大腸桿菌。第三型分泌系統透過使用運輸的毒性蛋白之針,使毒性蛋白由細菌細胞質經由該針至直接運輸至宿主細胞的細胞質。使用該針提供穿過包括革蘭式陰性菌及人細胞真核細胞膜的二層膜之多層膜的通路。尤其,在大腸桿菌菌株中,該針狀複體係由大腸桿菌分泌組分F(EscF)與接附於該針尖端的大腸桿菌分泌蛋白A(EspA)構成,形成大體上中空結構,供組分從細菌至宿主人體細胞的通路之用。此時,例如EspB之細菌蛋白可透過此管導入該宿主細胞。雖然EspB的生理學可能尚未被完全了解,但在「The Enteropathogenic E.coli effector EspB facilitates microvillus effacing and Antiphagocytosis by Inhibiting Myosin Function」一文中(於CELL HOSTS AND MICROBE,pp 383-392(2007)之中),描述EspB與肌凝蛋白結合,最終抑制人體免疫反應之吞噬作用。通常,肌凝蛋白蛋白質與肌動蛋白絲(actin filament)交互作用,以參與細胞過程,例如清除可能細菌病原體的吞噬作用。當大腸桿菌放出的EspB抑制各種肌凝蛋白蛋白質與肌動蛋白絲之間的交互作用而抑制吞噬作用,則導致嬰兒、孩童及成人的腹瀉或其他胃痛,發生有害症狀。 The secretion system of many Gram-negative pathogens is a type 3 secretion system, which includes the following bacteria: Shigella, Salmonella, Pseudomonas, and E. coli. The third type of secretion system uses the needle of the transported toxic protein to transport the toxic protein from the bacterial cytoplasm through the needle to the cytoplasm of the host cell. The needle is used to provide access through a multi-layered membrane including a two-layered membrane of gram-negative bacteria and human cell eukaryotic cell membranes. In particular, in E. coli strains, the acicular complex consists of E. coli secreted component F (EscF) and E. coli secreted protein A (EspA) attached to the tip of the needle, forming a generally hollow structure for the components The use of pathways from bacteria to host human cells. At this time, a bacterial protein such as EspB can be introduced into the host cell through this tube. Although the physiology of EspB may not be fully understood, in the article " The Enteropathogenic E. coli effector EspB facilitates microvillus effacing and Antiphagocytosis by Inhibiting Myosin Function " (in CELL HOSTS AND MICROBE, pp 383-392 (2007)) Describes that EspB binds to myosin and ultimately inhibits the phagocytosis of the human immune response. Generally, myosin proteins interact with actin filaments to participate in cellular processes, such as phagocytosis that clears possible bacterial pathogens. When EspB released by E. coli inhibits the interaction between various myosin proteins and actin filaments and inhibits phagocytosis, it can cause diarrhea or other stomach pains in infants, children, and adults, causing harmful symptoms.
乳鐵蛋白的其中一種抗入侵機轉係抑制EspB移位至人細胞中。尤其,一種機轉可包括抑制用於EspB自細菌移位的必要分泌結構形成。乳鐵蛋白可降解EspA,即負責用於移位Esp B至宿主細胞中的類管狀結構的蛋白質。當EspA可藉由乳鐵蛋白降解時,穿過人細胞膜的通口即不會產生,因此減輕由EspB進入人細胞的細胞質所產生的發病。此外,乳鐵蛋白亦可具有蛋白分解活性,造成EspB的降解。最後,乳鐵蛋白有效干擾針狀複體與病原體分泌系統的連接,同時降解造成包括胃腸痛及腹瀉之症狀的蛋白質。 Lactoferrin, one of the anti-invasion transgenic lines, inhibits the translocation of EspB into human cells. In particular, a mechanism may include inhibiting the formation of necessary secretory structures for the translocation of EspB from bacteria. Lactoferrin degrades EspA, a protein responsible for the tubular-like structure that is used to displace Esp B into host cells. When EspA can be degraded by lactoferrin, the passage through the human cell membrane will not be generated, thus reducing the incidence of EspB entering the cytoplasm of human cells. In addition, lactoferrin can also have proteolytic activity, causing the degradation of EspB. Finally, lactoferrin effectively interferes with the connection of the acicular complex to the pathogen's secretory system, while degrading proteins that cause symptoms including gastrointestinal pain and diarrhea.
提供下列實例以說明本文之營養組成物實施例,但不應解讀為對本文之任何限制。熟悉本技術領域者於考量本 文所揭露營養組成物或方法的說明書或實施,將清楚明白本文中之申請專利範圍範疇內的其他實施例。本說明書連同本實例僅欲視為例示,本文的範圍及精神係由接續於實例之後的申請專利範圍所指定。 The following examples are provided to illustrate the nutritional composition examples herein, but should not be construed as any limitation to this document. Those familiar with the technical field should consider this The description or implementation of the nutritional composition or method disclosed herein will clearly understand other embodiments within the scope of the patent application herein. This specification, together with this example, is only to be regarded as an example, and the scope and spirit of this document are specified by the scope of patent application following the example.
本實例示例單獨藉由乳鐵蛋白以及乳鐵蛋白合併聚葡萄糖及半乳糖寡醣病原體、即阪崎腸桿菌4603和29004以及大腸桿菌E2348/69的抑制作用。 This example exemplifies the inhibition of lactoferrin and lactoferrin combined with polydextrose and galactooligosaccharide pathogens, namely E. sakazakii 4603 and 29004, and E. coli E2348 / 69.
依初步實驗,測得阪崎腸桿菌4603和29004培養菌對HEp-2細胞產生最高的附著率。於培養6小時後,離心收集這些阪崎腸桿菌菌株培養菌及大腸桿菌E2348/69培養菌、以磷酸鹽緩衝液沖洗、並以補充10%胎牛血清的最低必需培養基(MEM)重新懸浮。HEp-2細胞(自ATTC取得)於含有25 ml之補充有10%FBS的MEM的75 cm2組織培養瓶、在CO2恆溫培養箱中於組織培養條件生長。藉由加入0.5 ml的0.25%胰蛋白酶-EDTA溶液(Sigma)並在組織培養條件培養共15分鐘,收集長滿(confluent)之HEp-2細胞。以0.5 ml的FBS將胰蛋白酶去活化,並在24孔組織培養盤中將細胞種於12-mm直徑的蓋玻片上,每孔約3.6 x 105活細胞。在開始各實驗前或達到長滿為止,將盤置於組織培養條件下共二天。 According to preliminary experiments, it was determined that E. sakazakii 4603 and 29004 cultured bacteria had the highest adhesion rate to HEp-2 cells. After 6 hours of incubation, these E. sakazakii strains and E. coli E2348 / 69 cultures were collected by centrifugation, rinsed with phosphate buffer, and resuspended in 10% fetal bovine serum-required minimal medium (MEM). HEp-2 cells (obtained from ATTC) were grown in 75 cm 2 tissue culture flasks containing 25 ml of MEM supplemented with 10% FBS in tissue culture conditions in a CO 2 incubator. Confluent HEp-2 cells were collected by adding 0.5 ml of a 0.25% trypsin-EDTA solution (Sigma) and culturing under tissue culture conditions for a total of 15 minutes. Trypsin was deactivated with 0.5 ml of FBS and cells were seeded on 12-mm diameter coverslips in a 24-well tissue culture plate with approximately 3.6 x 10 5 viable cells per well. Plates were placed under tissue culture conditions for two days before starting each experiment or until reaching fullness.
檢測即刻開始前,終濃度單獨為0.1 mg/ml、0.6 mg/ml及1 mg/ml之乳鐵蛋白及乳鐵蛋白合併1:1半乳糖 寡醣(自DOMO取得)及聚葡萄糖(自DMV取得)之摻合物至終濃度4 mg/ml及16 mg/ml係加入HEp-2細胞。亦製備未含乳鐵蛋白的對照孔。接著,900:1的大腸桿菌或阪崎腸桿菌培養菌(約含107細胞數)加入各孔中(三重覆組)。之後,組織培養盤置於37℃的CO2恆溫培養箱中共三小時。接以沖洗該些孔五次,以移除未附著的細胞,並藉由顯微鏡計數及定量即時PCR計算附著的細菌數。 Immediately before the test, lactoferrin and lactoferrin with final concentrations of 0.1 mg / ml, 0.6 mg / ml, and 1 mg / ml combined with 1: 1 galactooligosaccharide (obtained from DOMO) and polydextrose (from DMV (Obtained) was added to HEp-2 cells to a final concentration of 4 mg / ml and 16 mg / ml. Control wells without lactoferrin were also prepared. Subsequently, 900: 1, or E. sakazakii bacteria culture (number of cells containing about 107) were added to each well (repeated three groups). Thereafter, the tissue culture plate was placed in a CO 2 incubator at 37 ° C. for a total of three hours. The wells were then washed five times to remove unattached cells, and the number of attached bacteria was counted by microscope counting and quantitative real-time PCR.
針對顯微鏡計數,蓋玻片以100%甲醇固定、以10% Geimsa染色共15分鐘、以蒸餾水沖洗、並乾燥整夜。蓋玻片定著於顯微鏡載玻片上,並於位相差顯微鏡下以100x物鏡觀察。使用Motic Image軟體將各蓋玻片拍攝十五張顯微照片,接著為整個蓋玻片建立幾何圖案。使用Image J影像分析軟體計數各影像中的細胞及細菌數。附著性以每個HEp-2細胞的附著細菌數計算。附著性抑制係以對照組中每個細胞的附著細菌數減去處理組每個細胞的附著細菌數後再除以對照組中每個細胞的附著細菌數計算。針對使用大腸桿菌培養菌的實驗,人工計數具有大腸桿菌微菌落的細胞。有四或更多個細菌的細胞則認定為陽性(具有典型局部附著性狀)。測定具有附著微菌落的HEp-2細胞數,並如上所述計算附著性抑制。實驗以三重覆組進行,並重覆三次(n=9)。 For microscope counting, coverslips were fixed with 100% methanol, stained with 10% Geimsa for 15 minutes, rinsed with distilled water, and dried overnight. The coverslip was fixed on a microscope slide and observed under a phase-contrast microscope with a 100x objective lens. Fifteen photomicrographs were taken of each cover glass using Motic Image software, and then a geometric pattern was created for the entire cover glass. Image J software was used to count the number of cells and bacteria in each image. Adhesion was calculated as the number of attached bacteria per HEp-2 cell. Adhesion inhibition was calculated by subtracting the number of bacteria attached to each cell in the control group from the number of bacteria attached to each cell in the control group and then dividing by the number of bacteria attached to each cell in the control group. For experiments using E. coli cultures, cells with E. coli microcolonies were manually counted. Cells with four or more bacteria were considered positive (with typical local attachment traits). The number of HEp-2 cells with attached microcolonies was measured, and the inhibition of adhesion was calculated as described above. The experiment was performed in a triplicate group and repeated three times (n = 9).
除了顯微鏡計數之外,亦藉由定量即時PCR(qRT-PCR)計數被附著的細胞(如Humphries等人, Interactions of enteropathogenic Escherichia coli with pediatric and adult intestinal biopsy specimens during early adherence,Infect.Immun.,77,4463-4468(2009)所述)。簡言之,自經感染之HEp-2細胞抽取基因組DNA,並使用放大阪崎腸桿菌4603和29004或大腸桿菌2348/69的16s rRNA區域之寡核苷酸引子,以qRT-PCR定量。使用經適當稀釋的全基因組DNA作為內控制組,並製備qPCR終點相關於細胞濃度的標準曲線。該PCR混合物由11.25μl SYBR溶液、2.5 MasterMix、1 μl的各引子及5 μl的DNA模板組成。該PCR反應係使用Eppendorf Mastercycler Realplex2進行。 In addition to microscopic counting, attached cells are also counted by quantitative real-time PCR (qRT-PCR) (such as Humphries et al., Interactions of enteropathogenic Escherichia coli with pediatric and adult intestinal biopsy specimens during early adherence, Infect. Immun., 77, 4463-4468 (2009)). Briefly, genomic DNA was extracted from infected HEp-2 cells and quantified by qRT-PCR using oligonucleotide primers containing 16s rRNA regions of E. sakazakii 4603 and 29004 or E. coli 2348/69. An appropriately diluted whole genomic DNA was used as the internal control group, and a standard curve for qPCR endpoints as a function of cell concentration was prepared. The PCR mixture consisted of 11.25 μl SYBR solution, 2.5 MasterMix, 1 μl of each primer, and 5 μl of DNA template. This PCR reaction was performed using Eppendorf Mastercycler Realplex2.
本實例說明根據本文之營養產品的實施例。 This example illustrates an example of a nutritional product according to this document.
此實例說明根據本文的營養產品之另一實施例。 This example illustrates another embodiment of a nutritional product according to this document.
此實例說明可用於製備根據本文的營養產品之成分的一實施例。 This example illustrates one embodiment of the ingredients that can be used to prepare a nutritional product according to this document.
此實例說明可用於製備根據本文的營養產品之成分的另一實施例。 This example illustrates another example of ingredients that can be used to prepare a nutritional product according to this document.
所有本說明書中所引用之參考文獻、包括但不限於所有論文、刊物、專利、專利申請、發表、文字、報導、文稿、小冊、書籍、網路文章、期刊文章、定期性刊物等等,在此係以引用方式將它們全部併入本說明書。本文中之參考資料的討論僅欲用於總結其作者之主張,並不作為承認任何參考資料構成先前技術。申請人保留挑戰引用之參考文獻之準確性和針對性的權利。 All references cited in this specification, including but not limited to all papers, journals, patents, patent applications, publications, texts, reports, manuscripts, brochures, books, web articles, journal articles, periodic publications, etc., All of them are incorporated herein by reference. The discussion of references in this article is intended only to summarize the author's claims and is not an admission that any reference constitutes prior art. The applicant reserves the right to challenge the accuracy and relevance of the cited references.
雖然已使用特定術語、裝置及方法說明本文之較佳實施例,但此說明僅為描述目的。所使用文字為說明文字而非限制性文字。應了解,本技術領域中具有通常知識者在 未悖離於下列申請專利範圍所主張的本文之精神及範疇下,可進行更改及變動。此外,應了解,各種實施例可全部或部分相互取代。舉例而言,雖然已例示用於製造依據該些方法所製成的市售無菌液體營養補充物的方法,但其他使用亦被涵蓋。因此,後附申請專利範圍之精神及範疇不應受限於本說明中所含的較佳態樣。 Although specific terms, devices, and methods have been used to describe the preferred embodiments herein, this description is for illustrative purposes only. The text used is descriptive rather than restrictive. It should be understood that those with ordinary knowledge in the art Changes and modifications may be made without departing from the spirit and scope of this document as claimed in the following patent application scope. In addition, it should be understood that various embodiments may be substituted in whole or in part for each other. For example, although methods for making commercially available sterile liquid nutritional supplements made according to these methods have been exemplified, other uses are also covered. Therefore, the spirit and scope of the appended patent application scope should not be limited to the preferred aspects contained in this description.
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US12/980,813 US8648036B2 (en) | 2010-12-29 | 2010-12-29 | Use of nutritional compositions including lactoferrin and one or more prebiotics in inhibiting adhesion of pathogens in the gastrointestinal tract |
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US20140271978A1 (en) * | 2013-03-15 | 2014-09-18 | Mead Johnson Nutrition Company | Low-buffer nutritional compositions and uses thereof |
US9609888B2 (en) * | 2013-07-31 | 2017-04-04 | Mead Johnson Nutrition Company | Nutritional compositions containing synergistic combination and uses thereof |
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