TW201521603A - 包含具有抗發炎性質之肽組份之營養組成物及其用途 - Google Patents
包含具有抗發炎性質之肽組份之營養組成物及其用途 Download PDFInfo
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- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 1
- 235000019190 thiamine hydrochloride Nutrition 0.000 description 1
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- YXVCLPJQTZXJLH-UHFFFAOYSA-N thiamine(1+) diphosphate chloride Chemical compound [Cl-].CC1=C(CCOP(O)(=O)OP(O)(O)=O)SC=[N+]1CC1=CN=C(C)N=C1N YXVCLPJQTZXJLH-UHFFFAOYSA-N 0.000 description 1
- GUGWNSHJDUEHNJ-UHFFFAOYSA-N thiamine(1+) monophosphate chloride Chemical compound [Cl-].CC1=C(CCOP(O)(O)=O)SC=[N+]1CC1=CN=C(C)N=C1N GUGWNSHJDUEHNJ-UHFFFAOYSA-N 0.000 description 1
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Abstract
本發明關於包含蛋白質等效來源之營養組成物,其中20%至80%之該蛋白質等效來源包括含有SEQ ID NO 4、SEQ ID NO 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及SEQ ID NO 63之肽組份,且20%至80%之該蛋白質等效來源包含完整蛋白質、部分水解蛋白質或彼等之組合。本發明進一步關於經由提供該營養組成物予目標個體來減少發炎反應及/或製造促炎性細胞因子(即,介白素17)之方法。
Description
本發明關於包含肽組份之營養組成物,其中該肽組份包含選定之肽類的組合。於某些實施態樣中,當被投予個體時,該營養組成物減少發炎反應。更具體的說,本發明之營養組成物可減少促炎性細胞因子,諸如介白素-17(以下稱為“IL-17”)之製造。本文所描述之營養組成物可配製成用於成人及兒科個體。
細胞因子為由多種細胞類型分泌之細胞信號傳導、免疫調節蛋白質,且其廣泛用於細胞間之通訊。幾乎所有有核細胞,尤其是血管內皮細胞和上皮細胞,以及彼等之定居巨噬細胞為細胞因子之有效製造者。促炎性細胞因子包括介白素-12(“IL-12”)、介白素-11(“IL-11”)、介白素-17(“IL-17”)、介白素-18(“IL-18”)、介白素-5(“IL-5”)、介白素-4(“IL-4”)、干擾素-γ(“IFN-γ”)、介白素-8(“IL-8”)、腫瘤壞死因子-α(“TNF-α”)、腫瘤壞死因子-
β(“TNF-β”)、介白素-6(“IL-6”)、介白素-1(“IL-1”)、介白素-20(“IL-20”)、介白素-33(“IL-33”)、白血球抑制因子(“LIF”)、制瘤素M(“OSM”)、睫狀神經營養因子(“CNTF”)、轉化生長因子-β(“TGF-β”)、粒細胞-巨噬細胞群落(“GM-CSF”)。尤其,IL-17為一種作為延遲型反應中之傳介子的細胞因子,其係經由增加各種組織中趨化因子之製造來將單核細胞及嗜中性粒細胞招募至發炎部位。IL-17係由T-輔助細胞製造,且係由IL-23誘導,這導致延遲型反應中之破壞性組織損害。IL-17之功能係作為回應病原體入侵免疫系統之促炎性細胞因子,並引起病原體之細胞外基質被破壞。
據報導,IL-17族之細胞因子的免疫調節功能可能是由於IL-17細胞因子誘導許多免疫信號傳導分子之事實。IL-17涉及誘導及介導促炎性反應。此外,IL-17與過敏性反應有關,因其誘導許多其他細胞因子之製造,包括,但不限於IL-6、B-CSF、GM-CSF、IL-1β、TGF-β、TNF-α、趨化因子,包括IL-8、GRO-α和MCP-1,以及前列腺素,諸如PGE2。IL-17誘導各種各樣細胞類型,包括纖維母細胞、內皮細胞、上皮細胞、角質形成細胞及巨噬細胞製造這些促炎性化合物。
由於其誘導發炎反應之能力,IL-17和許多與免疫及自體免疫相關之疾病有關。由IL-17介導之發炎病況包括氣喘、過敏、皮膚病況,諸如皮膚炎、牛皮癬及濕疹、發炎性腸病和關節炎。用於治療自體免疫性疾病之傳
統藥物嘗試阻止涉及自體免疫攻擊的炎症。然而,這些藥物中有多種亦抑制免疫系統對抗感染之能力,且可能有許多嚴重的副作用。此外,該發炎反應在某些新生兒及兒童疾病的發病中扮演重要角色。
因此,能減少個體之發炎反應的安全但有效之營養組成物是有需要的。更具體的說,對於能減少嬰兒之發炎反應,從而協助減少某些新生兒及兒童疾病發病,但不會實質上改變免疫系統對抗感染的能力,且在食用時不會產生不欲有之副作用的嬰兒配方是有需要的。
簡單地說,於一實施態樣中,本發明針對包含碳水化合物來源、蛋白質等效來源及脂肪來源之營養組成物,其中該蛋白質等效來源包含包括來自下列表1之選定的肽類之肽組份。於一些實施態樣中,20%至80%之該蛋白質等效來源包括含有SEQ ID NO 4、SEQ ID NO 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及SEQ ID NO 63之肽組份;且20%至80%之該蛋白質等效來源包含完整蛋白質、部分水解蛋白質或彼等之組合。
於另一實施態樣中,該營養組成物包含碳水化合物來源、蛋白質等效來源及脂肪來源,其中該蛋白質等效來源包含至少3種選自由下列所組成之群組的肽類:
SEQ ID NO 4、SEQ ID NO 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,及至少5種選自表1之額外的肽類;且其中20%至80%之該蛋白質等效來源包含完整蛋白質、部分水解蛋白質或彼等之組合。
於某些實施態樣中,該營養組成物為兒科營養組成物(諸如嬰兒配方或成長奶)、成人營養組成物或營養補充劑。
於一些實施態樣中,本發明之營養組成物能夠減少個體之促炎性反應。例如,當被投予個體時,該營養組成物減少促炎性細胞因子,諸如介白素-12(“IL-12”)、IL-17、介白素-5(“IL-5”)、介白素-4(“IL-4”)、干擾素-γ(“IFN-γ”)、介白素-8(“IL-8”)、腫瘤壞死因子α(“TNF-α”)、介白素-6(“IL-6”)及介白素-1(“IL-1”)之製造。因此,於一些實施態樣中,本發明針對用於減少個體之促炎性反應的方法,其包含投予個體如本文所描述之營養組成物。
應理解的是,前文之一般描述及以下本發明實施態樣之詳細描述意欲提供用於理解所申請專利之揭示內容的性質和特性之綜述或框架。該描述係用於解釋所申請專利之題材的原理和操作。熟習本技藝之人士在閱讀下列揭露內容後將清楚明白本發明之其他及進一步的特性和優點。
第1圖說明由暴露於高度水解之酪蛋白的經活化之原代人樹突細胞分泌之促炎性細胞因子IL-12p70。
第2圖說明由暴露於高度水解之酪蛋白的大於500道耳吞之餾分的經活化之原代人樹突細胞分泌的促炎性細胞因子IL-12p70。
第3圖說明由暴露於高度水解之酪蛋白之經活化的人樹突細胞分泌之促炎性細胞因子干擾素-γ。
第4圖說明由暴露於高度水解之酪蛋白的大於500道耳吞之餾分之經活化的原代人樹突細胞分泌之促炎性細胞因子干擾素-γ。
第5圖說明由暴露於高度水解之酪蛋白之經活化的人樹突細胞分泌之促炎性細胞因子介白素-8。
第6圖說明由暴露於高度水解之酪蛋白的大於500道耳吞之餾分之經活化的原代人樹突細胞分泌之促炎性細胞因子介白素-8。
第7圖說明由暴露於高度水解之酪蛋白之經活化的人樹突細胞分泌之促炎性細胞因子腫瘤壞死因子-α。
第8圖說明由暴露於高度水解之酪蛋白的大於500道耳吞之餾分之經活化的原代人樹突細胞分泌之促炎性細胞因子腫瘤壞死因子-α。
第9圖說明由暴露於高度水解之酪蛋白之經活化的人樹突細胞分泌之促炎性細胞因子介白素-6。
第10圖說明由暴露於高度水解之酪蛋白的大於500道耳吞之餾分之經活化的原代人樹突細胞分泌之促炎性細胞因子介白素-6。
第11圖說明由暴露於高度水解之酪蛋白之經活化的人樹突細胞分泌之促炎性細胞因子介白素-1β。
第12圖說明由暴露於高度水解之酪蛋白的大於500道耳吞之餾分之經活化的原代人樹突細胞分泌之促炎性細胞因子介白素-1β。
第13圖顯示出暴露於高度水解之酪蛋白之經活化的原代人巨噬細胞分泌之腫瘤壞死因子-α。
第14圖說明當暴露於高度水解之酪蛋白的大於500道耳吞餾分時,經活化之原代人巨噬細胞分泌之腫瘤壞死因子-α。
第15圖說明NOD小鼠、BALB/C小鼠及被餵食高度水解之酪蛋白飲食的NOD小鼠的迴腸中分泌之介白素-1(IL-1)。
第16圖說明NOD小鼠、BALB/C小鼠及被餵食高度水解之酪蛋白飲食的NOD小鼠的迴腸中分泌之介白素-18(IL-18)。
第17圖說明NOD小鼠、BALB/C小鼠及被餵食高度水解之酪蛋白飲食的NOD小鼠的迴腸中分泌之介白素-6(IL-6)。
第18圖說明NOD小鼠、BALB/C小鼠及被餵食高度水解之酪蛋白飲食的NOD小鼠的迴腸中分泌之介
白素-17(IL-17)。
第19圖說明NOD小鼠、BALB/C小鼠及被餵食高度水解之酪蛋白飲食的NOD小鼠之腸引流淋巴結T細胞分泌之IFN-γ。
第20圖說明NOD小鼠、BALB/C小鼠及被餵食高度水解之酪蛋白飲食的NOD小鼠之腸引流淋巴結T細胞分泌之介白素-4。
第21圖說明NOD小鼠、BALB/C小鼠及被餵食高度水解之酪蛋白飲食的NOD小鼠之腸引流淋巴結T細胞分泌之介白素-17。
第22圖說明從暴露於高度水解之酪蛋白、未經無菌過濾之高度水解之酪蛋白及高度水解之酪蛋白的大於500道耳吞之餾分的小鼠巨噬細胞分泌之促炎性細胞因子介白素-12p40。
第23圖說明從暴露於高度水解之酪蛋白、未經無菌過濾之高度水解的酪蛋白及高度水解之酪蛋白的大於500道耳吞之餾分的小鼠巨噬細胞分泌之促炎性細胞因子介白素-1β。
第24圖說明從暴露於高度水解之酪蛋白、未經無菌過濾之高度水解的酪蛋白及高度水解之酪蛋白的大於500道耳吞之餾分的小鼠巨噬細胞分泌之促炎性細胞因子介白素-6。
第25圖說明從暴露於高度水解之酪蛋白、未經無菌過濾之高度水解的酪蛋白及高度水解之酪蛋白的大
於500道耳吞之餾分的小鼠巨噬細胞分泌之腫瘤壞死因子-α。
第26圖說明從暴露於高度水解之酪蛋白、未經無菌過濾之高度水解的酪蛋白及高度水解之酪蛋白的大於500道耳吞之餾分的小鼠巨噬細胞分泌之促炎性細胞因子介白素-12p40、介白素-1β、介白素-6、和腫瘤壞死因子-α。
現在參考本發明之實施態樣的詳細內容,下文中列舉其一或多個實例。所提供之各實例係用於解釋本發明之營養組成物,而非限制。事實上,熟習本技藝之人士將清楚明白本發明之教示內容可在不偏離本發明之範圍下做各種修改及變化。例如,所說明或描述之為一種實施態樣之一部分的特性可與另一實施態樣一起使用,以產生更進一步之實施態樣。
因此,本發明意圖涵蓋這類在所附之申請專利範圍及其同等項之範圍內的修改和變化。本發明之其他目的、特徵及態樣揭露於下列之詳細描述中或可從其中顯明。本技藝之一般技術人士將理解目前的討論僅用於說明示例性實施態樣,而不欲限制本發明之更廣泛的態樣。
“營養組成物”意指滿足個體之至少一部分營養需求的物質或調合物。在本發明之全文中,術語“營養”、“營養配方”、“腸內營養”及“營養補充劑”係作為營養組成
物之非限制性實例。再者,“營養組成物”可指液體、粉劑、凝膠、糊劑、固體、濃縮物、懸浮液或為即時使用形式之腸內配方、口服配方、嬰兒配方、兒科個體配方、兒童配方、成長奶及/或成人配方。
術語“腸內”意指可通過胃腸道或消化道,或在胃腸道或消化道內投遞。“腸內投予”包括經口餵食、胃內餵食、經幽門投予或任何其他進入消化道之投予。“投予”較“腸內投予”更廣義且包括胃腸道外投予或任何其他將物質投入個體內的投予途徑。
術語“醫療食品”係指被配製成或欲用於飲食管理疾病或失調的腸內組成物。醫療食品可能為用於口服或管飼(鼻胃管)之食物,其可被歸類為用於飲食管理具有獨特之營養需求的特定醫學失調、疾病或病況,且計劃在醫師之監督下使用。
本文所使用之術語“肽”係描述胺基酸之線性分子鏈,包括單鏈分子或其片段。本文所描述之肽類所包含的胺基酸總數不超過50個。肽類可進一步形成由至少兩個完全相同或不同之分子所組成之寡聚體或多聚體。此外,其中之胺基酸及/或肽鍵已被功能性類似物取代之這類肽的肽模擬物亦包含在術語“肽”中。這類功能性類似物可包括,但不限於除了該20種由基因編碼之胺基酸外的所有已知胺基酸,諸如硒代半胱胺酸。
術語“肽”亦可指經天然改質之肽類,其中該修改係藉由,例如糖基化、乙醯化、磷酸化及本技藝所周
知之類似修改進行。於一些實施態樣中,該肽組份與本文中亦揭示之蛋白質來源不同。此外,肽類可藉,例如重組、半合成、合成方法製造,或可從天然來源取得,諸如將蛋白質(包括,但不限於酪蛋白)水解後取得,這些均根據本技藝中已知之方法進行。
術語“水解度”係指肽鍵被水解方法打破之程度。例如,於一些實施態樣中,本發明之蛋白質等效來源可包含其水解度不大於40%之蛋白質。
術語“部分水解”意指其水解度大於0%,但小於約50%。
術語“高度水解”意指其水解度大於或等於約50%。
當用來指稱水解蛋白質或蛋白質水解產物時,術語“莫耳質量分佈”係關於存在於蛋白質水解產物中之每個肽的莫耳質量。例如,其莫耳質量分佈大於500道耳吞之蛋白質水解產物意指包含在該蛋白質水解產物中的每個肽之莫耳質量至少為500道耳吞。因此,於一些實施態樣中,表1及表2中所揭示之肽類係源自其莫耳質量分佈大於500道耳吞之蛋白質水解產物。為了製造其莫耳質量分佈大於500道耳吞之蛋白質水解產物,可將蛋白質水解產物進行某些過濾程序或任何其他本技藝中已知之用於去除莫耳質量小於500道耳吞之肽、胺基酸及/或其他蛋白物質的程序。為了本發明之目的,可使用本技藝中已知之任何方法來製造其莫耳質量分佈大於500道耳吞的蛋白
質水解產物。
術語“蛋白質等效物”或“蛋白質等效來源”包括任何蛋白質來源,諸如大豆、蛋、乳清或酪蛋白,以及非蛋白質來源,諸如肽或胺基酸。此外,該蛋白質等效來源可為任何本技藝中所使用者,例如脫脂乳、乳清蛋白、酪蛋白、大豆蛋白、水解蛋白質、肽、胺基酸及類似物。用於實行本發明之牛乳蛋白來源包括,但不限於乳蛋白粉、乳蛋白濃縮物、乳蛋白分離物、脫脂乳固體、脫脂乳、脫脂奶粉、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、酪蛋白酸鹽(例如酪蛋白酸鈉、酪蛋白酸鈉鈣、酪蛋白酸鈣)、大豆蛋白及彼等之任何組合。於一些實施態樣中,該蛋白質等效來源可包含水解蛋白質,包括部分水解蛋白質及高度水解蛋白質。於一些實施態樣中,該蛋白質等效來源可包括完整蛋白質。更特別地,該蛋白質等效來源可包括a)約20%至約80%之此交所描述的肽組份,及b)20%至80%之完整蛋白質、部分水解蛋白質或彼等之組合。
術語“蛋白質等效來源”亦包含游離胺基酸。於一些實施態樣中,該胺基酸可包含,但不限於組胺酸、異白胺酸、白胺酸、賴胺酸、蛋胺酸、半胱胺酸、苯丙胺酸、酪胺酸、蘇胺酸、色胺酸、纈胺酸、丙胺酸、精胺酸、天門冬醯胺、天門冬胺酸、麩胺酸、麩醯胺、甘胺酸、脯胺酸、絲胺酸、肉鹼、牛磺酸及彼等之混合物。於一些實施態樣中,該胺基酸可為支鏈型胺基酸。於某些其
他實施態樣中可包含小胺基酸肽類作為該營養組成物之蛋白質組份。這類小胺基酸肽類可為天然產生的或合成的。
“兒科個體”係指小於13歲的人。於一些實施態樣中,兒科個體係指介於出生和8歲之間的人個體。於其他實施態樣中,兒科個體係指介於1和6歲之間的人個體。再於進一步之實施態樣中,兒科個體係指介於6和12歲之間的人個體。術語“兒科個體”可指如下述之嬰兒(早產兒或足月)及/或兒童。
“嬰兒”意指年齡在出生到不超過一歲之範圍內的人個體且包括矯正年齡在0至12個月之嬰兒。“矯正年齡”一詞意指嬰兒之實足年齡減去嬰兒早產的時間量。因此,嬰兒若足月出生則矯正年齡為嬰兒的年齡。術語嬰兒包括低出生體重嬰兒、非常低出生體重嬰兒及早產兒。“早產兒”意指在妊娠第37週結束前出生的嬰兒。“足月兒”意指在妊娠第37週結束後出生之嬰兒。
“兒童”意指年齡在12個月至約13歲之範圍內的個體。於一些實施態樣中,兒童為年齡介於1歲和12歲之間的個體。於其他實施態樣中,術語“兒童們”或“兒童”係指介於1歲和約6歲之間,或介於約7歲和約12歲之間的個體。於其他實施態樣中,術語“兒童們”或“兒童”係指介於12個月和約13歲之間的任何年齡範圍。
“兒童營養產品”係指滿足兒童之至少一部分營養需求的組成物。成長奶為兒童營養產品之一種實例。
“嬰兒配方”意指滿足嬰兒之至少一部分營養需
求的組成物。在美國,嬰兒配方之內容係由21 C.F.R.章節100、106及107中所列出之聯邦法規規定。這些法規界定大量營養素、維生素、礦物質及其他成分之數量,以努力模擬人類母乳之營養及其他性質。
術語“成長奶”係指欲作為多樣化膳食之一部分,以支持年齡介於約1和約6歲之間的兒童正常生長和發育的廣大類別之營養組成物。
“營養完整”意指可作為唯一之營養來源的組成物,該組成物將實質上提供所有每日需要量之維生素、礦物質及/或微量元素,聯同蛋白質、碳水化合物及脂質。確切的說,“營養完整”係描述能提供支持個體正常生長和發育所需之足量的碳水化合物、脂質、必需脂肪酸、蛋白質、必需胺基酸、半必需胺基酸、維生素、礦物質及能源的營養組成物。
因此,顧名思義,對早產兒而言為“營養完整”之營養組成物將定性且定量地提供早產兒生長所需之足量的碳水化合物、脂質、必需脂肪酸、蛋白質、必需胺基酸、半必需胺基酸、維生素、礦物質及能量。
顧名思義,對足月兒而言為“營養完整”之營養組成物將定性且定量地提供足月兒生長所需之足量的全部碳水化合物、脂質、必需脂肪酸、蛋白質、必需胺基酸、半必需胺基酸、維生素、礦物質及能量。
顧名思義,對兒童而言為“營養完整”之營養組成物將定性且定量地提供兒童生長所需之足量的全部碳
水化合物、脂質、必需脂肪酸、蛋白質、必需胺基酸、半必需胺基酸、維生素、礦物質及能量。
當應用於營養素時,術語“必需”係指身體無法合成足夠用於正常生長及維持健康之量,因此必需由飲食供應的任何營養素。當應用於營養素時,術語“半必需”意指當身體無法取得足量之先質化合物以使內源性合成發生時,必需由膳食提供之營養素。
“益菌素”意指經由選擇性地刺激消化道中可改善宿主健康之一種或有限數量之細菌生長及/或活性而有益地影響該宿主的非消化性食物成分。
“益生菌”意指對宿主健康發揮至少一種有益效果之具有低或無致病性的微生物。
術語“經滅活之益生菌”意指其中所引用之益生有機體的代謝活性或繁殖能力已被降低或破壞的益生菌。然而,該“經滅活之益生菌”在細胞層級確實仍然保留至少一部分其生物乙二醇-蛋白質及DNA/RNA結構。如本文所使用之術語“經滅活”係與“無法存活”同義。更具體的說,一種非限制性之經滅活之益生菌的實例為經滅活之鼠李糖乳桿菌(Lactobacillus rhamnosus)GG(“LGG”)或“經滅活之LGG”。
除非另外具體指明,本文所使用之所有百分比、份數及比例係按總調合物之重量計。
本發明之營養組成物可能實質上不含任何本文所描述之可選擇或選定之成分,惟其該其餘的營養組成
物仍含有所有本文所描述之必要成分或特性。在此背景下,除非另外具體指明,術語“實質上不含”意指該選定之組成物中所含有之可選擇的成分可能少於該成分之功能量,通常少於0.1重量%,且亦包括含有0重量%之這類可選擇或選定之成分。
除非另外具體指明或明確暗示與所引用之背景相反,本發明中對單數特性或限制之所有引用應包括該對應之複數特性或限制,反之亦然。
除非另外具體指明或明確暗示與所引用之組合的背景相反,本文所使用之所有方法或加工步驟的組合可以任何順序執行。
本發明之方法和組成物(包括其組份)可包含此處所描述之實施態樣的必需要素和限制,以及本文所描述或可用於營養組成物中之任何額外或可選擇之成分、組份或限制,由彼等所組成或實質上由彼等所組成。
如本文所使用之術語“約”應解釋為係指被具體指定為終點之兩個數字內的任何範圍。對範圍之任何引用應被認為係提供對該範圍內之任何子集的支持。
促炎性反應或發炎係由某些刺激,諸如病原體、受損之細胞或刺激物引起。發炎反應可導致影響許多組織類型(包括上皮、結締組織、神經及腸)之各種失調。例如發炎反應可導致或加重皮膚病(諸如牛皮癬、濕疹及其他皮疹)、發炎性腸病、囊性纖維化、多發性硬化症、關節炎(例如類風濕關節炎)、過敏,甚至癌症之症
狀。發炎性疾病越來越普遍,許多甚至無法治愈。因此,提供可調節個體之促炎性反應的營養組成物將有利的。
因此,本發明大致上關於包含蛋白質等效來源之營養組成物,其中該蛋白質等效來源包括含有SEQ ID NO 4、SEQ ID NO 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及SEQ ID NO 63之肽組份。於一些實施態樣中,該肽組份可包含表1中所揭示之額外的肽類。例如,該組成物可包含至少10種表1中所揭示之額外的肽類。於一些實施態樣中,20%至80%之該蛋白質等效來源包含該肽組份,且20%至80%之該蛋白質等效來源包含完整蛋白質、部分水解蛋白質及彼等之組合。於一些實施態樣中,術語“額外”意指選擇與那些所列舉者不同的肽。
於另一實施態樣中,20%至80%之蛋白質等效來源包括含有至少3種選自由下列所組成之群組的肽組份:SEQ ID NO 4、SEQ ID NO 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,及至少5種選自表1之額外的肽類;且其中20%至80%之該蛋白質等效來源包含完整蛋白質、部分水解蛋白質或彼等之組合。
下列表1鑑別可包括在本發明之營養組成物的肽組份中之肽類的胺基酸序列。
下列表2進一步從表1鑑別可被包括在本發明之肽組份中的胺基酸序列之子集。
於一些實施態樣中,該營養組成物中之該肽組份的存在量為約0.2克/100千卡至約5.6克/100千卡。於其他實施態樣中,該營養組成物中之該肽組份的存在量為約1克/100千卡至約4克/100千卡。再於其他實施態樣中,該營養組成物中之該肽組份的存在量為約2克/100千卡至約3克/100千卡。
本發明之肽組份可與該營養組成物中之其他成分一起配製以提供該目標個體適當之營養素數量。於一些實施態樣中,該肽組份係包含在適合支持正常生長之營養完整的配方中。
於其他實施態樣中,該肽組份可包含可能添加在其他營養調合物(包括,但不限於食品及/或飲料)中之營養補充劑或添加劑。在本發明之目的方面,“營養補充劑”包括濃縮之營養素來源,例如本文所鑑別之肽類,或其他旨在用於補充正常飲食之具有營養或生理效果的物質。
該肽組份可以蛋白質等效來源之要素形式提供。於一些實施態樣中,表1及表2中所鑑別之肽類可由取自牛乳蛋白質之蛋白質等效來源提供,包括,但不限於牛酪蛋白及牛乳清。於一些實施態樣中,該蛋白質等效來源包含水解之牛酪蛋白或水解之牛乳清。因此,於一些實施態樣中,表1及表2中所鑑別之肽類可由酪蛋白水解產物提供。這類肽可藉由水解取得或可藉由熟習本技藝之人士已知之方法在體外合成。
本文中揭示一種水解方法之非限制性實施例。於一些實施態樣中,該方法可用於取得蛋白質水解產物及本發明之肽類。該蛋白質係使用蛋白水解酶,蛋白酶N水解。蛋白酶N“Amano”係購自美國伊利諾州Elgin,Amano Enzyme有限公司。蛋白酶N為源自枯草芽孢桿菌細菌菌種之蛋白水解酶製劑。該蛋白酶粉末之指定規格為“不少於150,000單位/克”意指一單位之蛋白酶N為在pH 7.0下,60分鐘內能產生等於100微克酪胺酸之胺基酸的酶量。為了製造本發明之嬰兒配方,蛋白酶N之使用量為水解之總蛋白質重量的約0.5%至約1.0%。
藉由蛋白酶N進行之蛋白質水解作用通常係在約50℃至約60℃之溫度下進行。令水解進行一段時間以取得介於約4%和10%之間的水解度。於一特定之實施態樣中,令水解進行一段時間以取得介於約6%和9%之間的水解度。於另一實施態樣中,令水解進行一段時間以取得約7.5%之水解度。此水解量可在約一個半小時至約3
小時之間取得。
水解期間應維持恆定之pH值。於本發明之方法中係將pH值調整且維持在介於約6.5和8之間。於一特定之實施態樣中係將pH值維持在約7.0。
為了維持乳清蛋白、酪蛋白、水及蛋白酶N之溶液的理想pH值,可在水解期間使用氫氧化鈉及/或氫氧化鉀之苛性鹼溶液來調節pH值。若使用氫氧化鈉來調整pH值,添加在該溶液中之氫氧化鈉的量應控制在使其包含之量少於該蛋白水解產物成品中之總固體的約0.3%。亦可使用10%氫氧化鉀溶液來將該溶液之pH值調節至所需之數值(無論是在添加酶之前或水解期間調節),以維持理想之pH值。
在蛋白質水解期間加入該溶液中之苛性鹼溶液的量可藉由pH恆穩器或經由持續及成比例地添加苛性鹼溶液來控制。該水解產物可藉由標準分批生產過程或藉由連續生產過程製造。
為了更能確保該蛋白質部分水解產物之一貫品質,將該水解產物進行酶去活化,以結束該水解過程。該酶去活化步驟可包括在約82℃之溫度下熱處理約10分鐘。或者,可經由將溶液在約92℃之溫度下加熱約5秒來將該酶去活化。當酶之去活化完成後,可將該水解產物以液體狀態儲存在低於10℃之溫度下。
於一些實施態樣中,該蛋白質等效來源包含水解蛋白質(包括部分水解蛋白質及高度水解蛋白質),
諸如酪蛋白。於一些實施態樣中,蛋白質等效來源包含水解蛋白質,包括其莫耳質量分佈大於500道耳吞之肽類。於一些實施態樣中,該水解蛋白質包含其莫耳質量分佈在約500道耳吞至約1500道耳吞之範圍內的肽類。再者,於一些實施態樣中,該水解蛋白質可包含其莫耳質量分佈在約500道耳吞至約2000道耳吞之範圍內的肽類。
於一些實施態樣中,該蛋白質等效來源可包含該肽組份、完整蛋白質、水解之蛋白質(包括部分水解蛋白質),及彼等之組合。於一些實施態樣中,20%至80%之該蛋白質等效來源包含本發明之肽組份。於一些實施態樣中,30%至60%之該蛋白質等效來源包含本發明之肽組份。再於其他實施態樣中,40%至50%之該蛋白質等效來源包含該肽組份。
於一些實施態樣中,20%至80%之該蛋白質等效來源包含完整蛋白質、部分水解蛋白質或彼等之組合。於一些實施態樣中,40%至70%之該蛋白質等效來源包含完整蛋白質、部分水解蛋白質或彼等之組合。再於進一步之實施態樣中,50%至60%之該蛋白質等效來源包含完整蛋白質、部分水解蛋白質或彼等之組合。
於一些實施態樣中,該蛋白質等效來源包含其水解度低於40%之部分水解蛋白質。於其他實施態樣中,該蛋白質等效來源可包含其水解度低於25%,或低於15%之部分水解蛋白質。
於一些實施態樣中,該營養組成物每100千
卡包含介於約1克和約7克之間的蛋白質等效來源。於其他實施態樣中,該營養組成物每100千卡包含介於約3.5克和約4.5克之間的蛋白質等效來源。
此外,包含該肽組份之蛋白質等效來源可藉由本技藝所熟知之任何方法添加入或納入該營養組成物中。於一些實施態樣中,可將該肽組份添加在營養組成物中,以補充該營養組成物。例如,於一實施態樣中,可將該肽組份添加在市售之嬰兒配方中。例如,Enfalac、Enfamil®、Enfamil®早產兒配方、帶有鐵之Enfamil®、Enfamil®LIPIL®、Lactofree®、Nutramigen®、Pregestimil®及ProSobee®(可從美國印第安納州,Evansville,Mead Johnson公司取得)可被輔以合適數量之肽組份並用於實行本發明。
本發明之包含肽組份的營養組成物可在一或多個劑量中每日投予。任何口服上可接受之劑量型均在本發明之考慮內。這類劑量型之實例包括,但不限於丸劑、片劑、膠囊、軟凝膠、液體、液體濃縮物、粉劑、酏劑、溶液、懸浮液、乳劑、錠劑、小珠、扁囊劑及彼等之組合。
於一些實施態樣中,可將該肽組份添加在更完整之營養產品中。於此實施態樣中,該營養組成物可含有蛋白質、脂肪及碳水化合物組份,並可用來補充飲食或可作為唯一之營養來源。
於一些實施態樣中,該營養組成物包含至少
一種碳水化合物來源。該碳水化合物來源可為任何本技藝所使用者,例如乳糖、葡萄糖、果糖、玉米糖漿固體、麥芽糊精、蔗糖、澱粉、稻米糖漿固體及類似物。在該營養組成物中之碳水化合物組份之量通常可在約5克/100千卡和約25克/100千卡之間變化。於一些實施態樣中,該碳水化合物之量係介於約6克/100千卡和約22克/100千卡之間。於其他實施態樣中,該碳水化合物之量係介於約12克/100千卡和約14克/100千卡之間。於一些實施態樣中,玉米糖漿固體為較佳者。再者,水解、部分水解及/或高度水解之碳水化合物可能較利於被納入該營養組成物中,因為它們容易消化。具體而言,水解碳水化合物較不可能含有過敏性表面結合部位。
適合用於本發明之碳水化合物物質的非限制性實例包括水解的或完整的、天然或經化學改質之來自玉米、木薯、稻米或馬鈴薯之糯和非糯形式澱粉。合適之碳水化合物的非限制性實例包括各種被表述為水解之玉米澱粉、麥芽糊精、麥芽糖、玉米糖漿、右旋糖、玉米糖漿固體、葡萄糖和各種其他葡萄糖聚合物,及彼等之組合的水解澱粉。其他合適之碳水化合物的非限制性實例包括那些常稱為蔗糖、乳糖、果糖、高果糖玉米糖漿、不可消化之寡醣(諸如果寡醣)者及彼等之組合。
於一些實施態樣中,該營養組成物可不含蛋白質且含有游離胺基酸作為該蛋白質等效來源之要素。於一些實施態樣中,該胺基酸可為支鏈胺基酸。於某些其他
實施態樣中,可包含小胺基酸肽類作為營養組成物之蛋白質組份。這類小胺基酸肽類可為天然產生或合成的。在該營養組成物中之游離胺基酸的量可在約1克/100千卡至約5克/100千卡之間變化。
該營養組成物亦可包含脂肪來源。用於本發明之營養組成物的合適脂肪或脂質來源可為任何本技藝中所已知或使用者,包括,但不限於動物來源,例如乳脂肪、奶油、奶油脂肪,蛋黃脂質;海洋來源,諸如魚油、海洋油、單細胞油;蔬菜和植物油,諸如玉米油、芥花油、葵花油、大豆油、棕櫚油、椰子油、高油酸葵花油、月見草油、菜籽油、橄欖油、亞麻仁(亞麻籽)油、棉籽油、高油酸紅花油、棕櫚油硬脂、棕櫚仁油、小麥胚芽油;中鏈甘油三酯油和脂肪酸之乳劑和酯類;以及彼等之任何組合。
於一些實施態樣中,該營養組成物包含約1.3克/100千卡至約7.2克/100千卡之脂肪來源。於其他實施態樣中,該脂肪來源之存在量可為約2.5克/100千卡至約6.0克/100千卡。再於其他實施態樣中,該脂肪來源之存在量可為約3.0克/100千卡至約4.0克/100千卡。
本發明之營養組成物可含有長鏈多元不飽和脂肪酸(“LCPUFA”)來源。合適之LCPUFA包括,但不限於DHA、二十碳五烯酸(“EPA”)、ARA、n-6通路中之亞油酸(18:2 n-6)、γ-亞麻酸(18:3 n-6)、二高-γ-亞麻酸(20:3 n-6)、α-亞麻酸(18:3 n-3)、十八碳
四烯酸(18:4 n-3)、二十碳四烯酸(20:4 n-3)、二十碳五烯酸(20:5 n-3)及二十二碳五烯酸(22:6 n-3)。
該營養組成物中之LCPUFA的量為至少約5毫克/100千卡,且可在約5毫克/100千卡至約100毫克/100千卡之間變化,更佳為約10毫克/100千卡至約50毫克/100千卡。
LCPUFA之來源包括乳製品,像蛋及乳脂;海洋油,諸如鱈魚、鯡魚、沙丁魚、鮪魚及許多其他魚類;某些動物脂肪、豬油、牛脂及微生物油,諸如真菌及海藻油,或者可為任何其他可取得LCPUFA並將其用於營養組成物中之經強化或非經強化形式的來源。該LCPUFA可為複雜混合物之一部分,其係藉由本技藝已知之旨在富集這類混合物中之LCPUFA及LCPUFA的衍生物或先質之分離技術取得。
該LCPUFA可以下列形式提供在營養組成物中:游離脂肪酸之酯;甘油單酯、甘油二酯及甘油三酯;磷酸甘油酯,包括卵磷脂;及/或彼等之混合物。此外,LCPUFA可以磷脂之形式(尤其是磷脂醯膽鹼)提供在營養組成物中。
於一實施態樣中,尤其是若該營養組成物為嬰兒配方時,該營養組成物係經輔以DHA和ARA二者。於此實施態樣中,該ARA:DHA之重量比可介於約1:3和約9:1之間。於一特殊之實施態樣中,該ARA:DHA
之重量比為約1:2至約4:1。
於一些實施態樣中,該營養組成物中之DHA的存在量較有利為至少約17毫克/100千卡,且可在約5毫克/100千卡至約75毫克/100千卡之間變化。於一些實施態樣中,DHA之存在量為約10毫克/100千卡至約50毫克/100千卡。
該營養組成物可使用本技藝中已知之標準技術被輔以含有DHA及/或ARA之油。例如,可經由取代等量之通常存在於該組成物中的油(諸如高油酸葵花油)來將DHA及ARA添加在該組成物中。另一實例為該含有DHA和ARA之油可經由取代等量之通常存在於不含DHA和ARA之組成物中的全部脂肪摻合物之其餘部分來添加在組成物中。
若使用DHA及/或ARA時,該DHA及/或ARA之來源可為本技藝中已知之任何來源,諸如海洋油、魚油、單細胞油、蛋黃脂質及腦脂質。於一些實施態樣中,該DHA和ARA係源自單細胞Martek油、DHASCO®及ARASCO®或彼等之變體。該DHA和ARA可為天然形式,惟其該LCPUFA來源之其餘部分不會對嬰兒造成任何實質之有害影響。或者,可使用精製形式之DHA和ARA。
於一實施態樣中,DHA及ARA之來源為如美國專利案第5,374,567;5,550,156;及5,397,591號(其揭示內容全文以引用方式併入本文)中所教示之單細胞油。
然而,本發明並不僅限於這類油。
此外,該營養組成物之一些實施態樣可模擬人類母乳之某些特徵。然而,為了滿足一些個體的特定營養需求,該營養組成物可包含較人乳所含有者更多量之某些營養組份。例如,該營養組成物可包含較人類母乳所含有者多量之DHA。因此,該營養組成物中增加的DHA含量可以彌補現有之DHA的營養不足。
於某些實施態樣中,該營養組成物亦可含有一或多種益菌素(亦稱為益菌素來源)。益菌素可刺激攝入之益生菌微生物的生長及/或活性、選擇性地減少在腸道中發現之病原體及有利地影響腸道中之短鏈脂肪酸的變化形廓。這類益菌素可為天然產生的、合成的或透過有機體及/或植物之遺傳工程操作研發,無論這類新來源為現在已知的或稍後研發的。可用於本發明中之益菌素可包括寡醣、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素。
更具體的說,可用於本發明中之益菌素可包括聚右旋糖、聚右旋糖粉、乳果糖、乳果寡醣、棉籽糖、葡萄寡醣、菊糖、果寡醣、異麥芽寡醣、大豆寡醣、乳果寡醣、木寡醣、殼寡醣(chito-oligosaccharide)、甘露寡醣、阿拉伯寡醣、唾液酸寡醣、岩藻寡醣(fuco-oligosaccharide)、半乳寡醣及龍膽寡醣(gentio-oligosaccharides)。於一些實施態樣中,存在於該營養組成物中之益菌素的總量可為約0.1克/100千卡至約1克
/100千卡。於某些實施態樣中,存在於該營養組成物中之益菌素的總量可為約0.3克/100千卡至約0.7克/100千卡。再者,該營養組成物可包含含有聚右旋糖(“PDX”)及/或半乳寡醣(“GOS”)之益菌素組份。於一些實施態樣中,該益菌素組份包含至少20%之GOS、PDX或彼等之混合物。
於一實施態樣中,若該益菌素組成物中使用PDX,該營養組成物中之PDX的量可在約0.1克/100千卡至約1克/100千卡的範圍內。於另一實施態樣中,該聚右旋糖的量係在約0.2克/100千卡至約0.6克/100千卡的範圍內。再於其他實施態樣中,該營養組成物中之PDX的量可為約0.1毫克/100千卡至約0.5毫克/100千卡。
於一實施態樣中,若該益菌素組成物中使用GOS,該營養組成物中之GOS的量可為約0.1克/100千卡至約1克/100千卡。於另一實施態樣中,該營養組成物中之GOS的量可為約0.2克/100千卡至約0.5克/100千卡。於其他實施態樣中,該營養組成物中之GOS的量可為約0.1毫克/100千卡至約1.0毫克/100千卡,或約0.1毫克/100千卡至約0.5毫克/100千卡。
於該營養組成物之特定的實施態樣中係將PDX與GOS組合投予。於本實施態樣中可將PDX和GOS以介於約9:1和1:9之間的PDX:GOS比率投予。於另一實施態樣中,該PDX:GOS之比率可介於約5:1和1:5之間。再於另一實施態樣中,該PDX:GOS之比率
可介於約1:3和3:1之間。於特定之實施態樣中,該PDX:GOS之比率可為約5:5。於另一特定之實施態樣中,該PDX對GOS之比率可為約8:2。
於特定之實施態樣中係將總量為至少約0.2毫克/100千卡或約0.2毫克/100千卡至約1.5毫克/100千卡之GOS和PDX補充在該營養組成物中。於一些實施態樣中,該營養組成物可包含總量為約0.6至約0.8毫克/100千卡之GOS及PDX。
於一些實施態樣中,該營養組成物可含有一或多種益生菌。本實施態樣可接受本技藝中已知之任何益生菌。於特定之實施態樣中,該益生菌可選自以下群組中之任何益生菌:乳酸桿菌種(Lactobacillus species)、鼠李糖乳桿菌GG(Lactobacillus rhamnosus GG)(ATCC編號53103)、雙岐桿菌種(Bifidobacterium species)、長雙岐桿菌BB 536(Bifidobacterium longum BB 536)(BL999,ATCC:BAA-999)、長雙岐桿菌AH1206(NCIMB:41382)、短雙岐桿菌AH1205(Bifidobacterium breve AH1205)(NCIMB:41387)、嬰兒雙岐桿菌35624(Bifidobacterium infantis 35624)(NCIMB:41003)及動物雙歧桿菌乳亞種BB-12(Bifidobacterium animalis subsp.lactis BB-12)(DSM編號10140)或彼等之任何組合。
若包含在組成物中,該益生菌之量可在每100千卡約含1×104至約1.5×1010cfu之益生菌之間變化,更佳為每100千卡約含1×106至約1×109cfu之益生菌。於
某些其他實施態樣中,該益生菌之量可在約1×107cfu/100千卡至約1×108cfu/100千卡之間變化。
於一實施態樣中,該益生菌可為可存活的或無法存活的。如本文所使用之術語“可存活的”係指活的微生物。術語“無法存活的”或“無法存活的益生菌”係指無法存活之益生菌微生物、彼等之細胞組份及/或彼等之代謝物。這類無法存活之益生菌可能已被熱滅殺或以其他方式滅活,但其保留能有利地影響該宿主之健康的能力。可用於本發明中之益生菌可為天然產生的、合成的或透過有機體之遺傳工程操作研發的,無論這類新來源為現在已知的或稍後研發的。
於一些實施態樣中,除了益生菌本身以外,本發明之營養組成物可包含來自益生菌分批培養過程之後期指數生長期的培養上清液(下文中稱為“培養上清液”);於特定之實施態樣中,該益生菌為LGG。分批培養之培養上清液(其亦可稱為“耗盡培養基”)可具有對抗病原體感染之保護作用,包括由阪崎克羅諾桿菌(C.sakazakii)造成之感染。具體的說,該收穫之培養上清液可以防止阪崎克羅諾桿菌入侵器官(諸如腦),並降低與阪崎克羅諾桿菌相關之死亡率。
於一些實施態樣中,該營養組成物包含用於治療或預防病原體感染之來自益生菌分批培養過程之後期指數生長期的培養上清液。於某些實施態樣中,該益生菌為LGG,且該病原體為阪崎克羅諾桿菌。
不欲受理論之限制,咸信,該培養物上清液之活性可歸因於該被發現在LGG之分批培養的後期指數期(或“對數期”)釋入培養基中之組份(包括蛋白質物質,且可能包括(胞外)多醣物質)的混合物。咸信,該培養上清液中之化學組成為數種胺基酸、寡肽和多肽,以及具各種分子量之蛋白質的混合物。該培養上清液可進一步包含多醣結構物及/或核苷酸。於一些實施態樣中,該培養上清液涉及整體,即,未分餾之培養上清液。進一步說,於一些實施態樣中,該培養上清液涉及整體,即,未分餾之培養上清液。
在細菌之分批培養中所認定之階段為熟習本技藝之人士所已知。這些階段為“滯後(lag)”、“對數(log)”(“對數(logarithmic)”或“指數(exponential)”)及“靜止(stationary)”和“死亡(death)”(或“對數下降(logarithmic decline)”)期。在存有活菌的所有時期中,該細菌代謝來自培養基之營養素並分泌(施加、釋出)物質入培養基中。該在生長階段之指定時點所分泌出之物質的組成一般而言是無法預測的。
於一些實施態樣中,根據該揭示內容及/或其實施態樣之組成物可藉由包含以下步驟之過程取得:(a)使用分批過程將益生菌,諸如LGG培養在合適之培養基中;(b)在該培養步驟之後期指數生長期收穫該培養上清液,該後期指數生長期之界定係參考介於該分批培養過程之滯後期及靜止期之間的後半段時間;(c)可選
擇地從該上清液除去低分子量成分,以保留分子量大於5千道耳吞(kDa),或甚至大於6kDa之成分;(d)從該培養上清液去除液體內容物,從而取得該組成物。
於本發明及其實施態樣中,分泌之物質係從後期指數期收穫。後期指數期係發生在中期指數期(其為指數期持續期間之一半時間,因此,對後期指數期之引用係指介於滯後期和靜止期之間的時間的後半段)後之時間。尤其是,本文所使用之術語“後期指數期”係指分批培養過程中介於滯後期和靜止期之間的時間的後四分之一部分。於本發明之一些實施態樣中係在指數期持續期間的75%至85%之時間點處收穫該培養上清液,最佳地,在指數期約經過六分之五的時間收穫。
術語“培養(cultivation或culturing)”係指在合適之培養基上或培養基中繁殖微生物(本例中之微生物為LGG)。這類培養基可具有多種類型,尤其是如本技藝中慣常使用之液體肉湯。較佳之肉湯為,例如一般用於培養乳桿菌之MRS肉湯。MRS肉湯通常包含聚山梨醇酯、醋酸鹽、鎂及錳(其已知可作為用於乳桿菌之特殊生長因子)及豐富營養基。典型之組成物包含(含量以克/升表示):來自酪蛋白之腖10.0;肉萃取物8.0;酵母萃取物4.0;D(+)-葡萄糖20.0;磷酸氫二鉀2.0;吐溫®80 1.0;檸檬酸三銨2.0;醋酸鈉5.0;硫酸鎂0.2;硫酸錳0.04。
於一些實施態樣中,本發明之培養上清液可被包含在為嬰兒配方之營養組成物中。收穫分泌出之細菌
產物帶來的問題是該培養基無法輕易地去除不欲有之組份。具體地說,這關係到用於相對弱勢之個體的營養產品,諸如嬰兒配方或臨床營養。若先將來自培養上清液之特定組份分離出、純化,再應用在營養產品中則此問題不會產生。然而,使用更完整之培養上清液是有需要的。此將用來提供能更佳地反映該益生菌(即,LGG)之天然作用的組成物。然而,人們不能只使用培養上清液本身作為用於欲特定用在嬰兒配方及類似物中之無法存活的益生菌物質之底質。
於一些實施態樣中,從LGG培養中收穫之培養上清液不含有營養組成物(諸如嬰兒配方)中所不欲有或不被普遍接受之組份(如可能存在於培養基中者)。在慣常存在於MRS肉湯中之聚山梨醇酯方面,用於培養細菌之培養基可包含乳化之非離子性表面活性劑,例如以聚乙氧基化山梨醇酐及油酸為基礎(通常可以吐溫®聚山梨醇酯之形式取得,諸如吐溫®80)。雖然食品(例如冰淇淋)中常發現這些表面活性劑在且通常被認為是安全的,但他們在食品管控中並不全被認為是令人滿意,甚至可被接受用於相對弱勢之個體的營養產品中(諸如嬰兒配方或臨床營養)。
因此,於一些實施態樣中,本發明採用其中可避免上述之聚山梨醇酯的培養基。為此,本發明之培養基不含有聚山梨醇酯,諸如吐溫80。於本發明之較佳實施態樣及/或其實施態樣中,該培養基可包含選自由下列
所組成之群組的油成分:油酸、亞麻籽油、橄欖油、菜籽油、葵花油及彼等之混合物。可理解的是,若實質上或完全避免存有聚山梨醇酯表面活性劑則可取得該油成分的全部益處。
於一些實施態樣中,該培養上清液可具有中性pH值,諸如介於pH 5和pH 7之間的pH值,較佳為pH 6。
除了前述者外,應注意的是乳桿菌(包括LGG)之分批培養為熟習本技藝之人士可取得之公知常識。因此,這些方法不需要在此進一步解釋。本發明之培養上清液可藉由任何已知之用於從細菌培養物分離培養上清液之技術收穫。這類技術為本技藝中所周知且包括,例如離心、過濾、沉降及類似技術。
本發明之營養組成物可以本技藝中已知之任何形式提供,諸如粉劑、凝膠、懸浮液、糊劑、固體、液體、液體濃縮物、可重構成之奶粉替代品或即時使用之產品。於某些實施態樣中,該營養組成物可包含營養補充劑、兒童營養產品、嬰兒配方、人乳強化劑、成長奶或任何其他經過設計之用於嬰兒或兒科個體的營養組成物。本發明之營養組成物包括,例如經口攝入,促進健康之物質,包括,例如食品、飲料、片劑、膠囊及粉劑。再者,本發明之營養組成物可被標準化成特定之卡路里含量,其可以即時使用之產品形式提供或可以濃縮形式提供。於一些實施態樣中,該營養組成物為粉劑形式,其粒徑係在5
微米至1500微米之範圍內,更佳為在10微米至300微米之範圍內。
若該營養組成物為即時使用產品之形式,該營養組成物之滲透重量莫耳濃度可介於約100和約1100毫滲透重量莫耳濃度/公斤水之間,更典型為約200至約700毫滲透莫耳濃度/公斤水。
於某些實施態樣中,該營養組成物為低過敏性。於其他實施態樣中,該營養組成物為符合猶太教規的食品(kosher)及/或符合穆斯林教規的食品(halal)。再於進一步之實施態樣中,該營養組成物含有非經遺傳工程改質之成分。於一實施態樣中,該營養調合物不含蔗糖。該營養組成物亦可不含乳糖。於其他實施態樣中,該營養組成物不含有任何中鏈甘油三酯油。於一些實施態樣中,該組成物中不含有角叉菜膠。於其他實施態樣中,該營養組成物不含有所有樹膠。
本發明之營養組成物並不限於包含本文中具體表列之營養素的組成物。任何營養素均可以該組成物之一部分的形式投遞,以滿足個體之營養需求及/或優化個體之營養狀態。
再者,於一些實施態樣中,該營養組成物為營養完整的,其含有欲作為個體唯一之營養來源的合適類型及量之脂質、碳水化合物、蛋白質、維生素及礦物質。確切的說,該營養組成物可選擇地包含任意數量之蛋白質、肽、胺基酸、脂肪酸、益生菌及/或彼等之代謝副產
物、益菌素、碳水化合物及任何可提供個體許多營養和生理助益之其他營養素或其他化合物。進一步說,本發明之營養組成物可包含香料、香料增强劑、甜味劑、色素、維生素、礦物質、治療成分、功能性食品成分、食品成分、加工成分或彼等之組合。
本發明之營養組成物可被標準化成特定之卡路里含量,其可以即時使用之產品形式提供,或者其可以濃縮形式提供。
於一些實施態樣中,本發明之營養組成物為成長奶。成長奶為欲用於1歲以上(通常為1-3歲、4-6歲、或1-6歲)之兒童的經強化之以乳為底質的飲品。其不為醫療食品,且不欲作為代餐或補充劑來解決特殊之營養缺乏。相反地,成長奶之設計係欲作為多樣化飲食的補充劑,以提供兒童達到持續性每日攝入所有必需維生素和礦物質、大量營養素加上額外之功能性膳食組份(諸如那些具有聲稱之促進健康特性的非必需營養素)的額外保障。
根據在地法規及所欲族群之膳食攝入信息,根據本發明之營養組成物的確切組成在各市場之間可有所變化。於一些實施態樣中,根據本發明之營養組成物係由乳蛋白來源(諸如全脂或脫脂奶),加上用於取得所需之感官性質之添加的糖和甜味劑,以及添加的維生素和礦物質所組成。該脂肪組成物通常係源自乳汁原料。總蛋白之目標可設定為符合人乳、牛乳之數值或較低之數值。總碳
水化合物之目標通常設定為提供盡可能少之添加的糖(諸如蔗糖或果糖)來取得可接受的味道。通常,維生素A、鈣及維生素D之添加量為符合區域牛奶的營養貢獻。另外,於一些實施態樣中,維生素和礦物質之添加量可為每一份量提供約20%之膳食參考攝入量(DRI)或20%之日需值(DV)。再者,根據經鑑定之所欲族群的營養需求、原料貢獻和地區規定,不同市場間之營養數值可有所變化。
該營養組成物中亦可添加一或多種足量之維生素及/或礦物質以供應個體之每日營養需求。本技藝之一般技術人士應理解維生素及礦物質需求將根據,例如兒童的年齡而有所不同。例如,嬰兒之維生素和礦物質需求可能與年齡介於1和13歲之間的兒童不同。因此,本實施態樣並不欲將該營養組成物限制在特定之年齡組,而是提供可接受之維生素和礦物質組份的範圍。
於提供用於兒童之營養組成物的實施態樣中,該組成物可選擇地包括,但不限於一或多種下列維生素或彼等之衍生物:維生素B1(硫胺素、焦磷酸硫胺素、TPP、三磷酸硫胺素、TTP、鹽酸硫胺素、單硝酸硫胺素)、維生素B2(核黃素、黃素單核苷酸、FMN、黃素腺嘌呤二核苷酸、FAD、乳黃素(lactoflavin)、卵黃素(ovoflavin))、維生素B3(菸鹼酸(niacin)、菸鹼酸(nicotinic acid)、菸醯胺(nicotinamide)、菸鹼醯胺(niacinamide)、菸醯胺腺嘌呤二核苷酸、NAD、菸鹼酸
單核苷酸、NicMN、吡啶-3-羧酸)、維生素B3先質色胺酸、維生素B6(吡哆醇、吡哆醛、吡哆胺、鹽酸吡哆醇)、泛酸(泛酸鹽、泛醇)、葉酸鹽(葉酸、葉酸(folacin)、蝶醯麩胺酸(pteroylglutamic acid))、維生素B12(鈷胺素、甲鈷胺素、脫氧腺苷鈷胺素、氰鈷胺素、羥鈷胺素、腺苷鈷胺素)、生物素、維生素C(抗壞血酸)、維生素A(視黃醇、醋酸視黃酯、棕櫚酸視黃酯、具其他長鏈脂肪酸之視黃酯、視黃醛、視黃酸、視黃醇酯)、維生素D(骨化醇、膽骨化醇、維生素D3、1,25-二羥基維生素D)、維生素E(α-生育酚、醋酸α-生育酚、琥珀酸α-生育酚、菸鹼酸α-生育酚、α-生育酚)、維生素K(維生素K1、葉綠醌、萘醌、維生素K2、甲萘醌(menaquinone)-7、維生素K3、甲萘醌-4、甲萘醌(menadione)、甲萘醌-8、甲萘醌-8H、甲萘醌-9、甲萘醌-9H、甲萘醌-10、甲萘醌-11、甲萘醌-12、甲萘醌-13)、膽鹼、肌醇、β-胡蘿蔔素及彼等之任何組合。
於提供兒童營養產品(諸如成長奶)的實施態樣中,該組成物可選擇地包括,但不限於一或多種下列礦物質或彼等之衍生物:硼、鈣、醋酸鈣、葡萄糖酸鈣、氯化鈣、乳酸鈣、磷酸鈣、硫酸鈣、氯化物、鉻、氯化鉻、吡啶甲酸鉻、銅、硫酸銅(copper sulfate)、葡萄糖酸銅、硫酸銅(cupric sulfate)、氟化物、鐵、羰基鐵、三價鐵、富馬酸亞鐵、正磷酸鐵、研磨鐵、多醣鐵、碘化物、碘、鎂、碳酸鎂、氫氧化鎂、氧化鎂、硬脂酸鎂、硫
酸鎂、錳、鉬、磷、鉀、磷酸鉀、碘化鉀、氯化鉀、醋酸鉀、硒、硫、鈉、多庫酯鈉、氯化鈉、硒酸鈉、鉬酸鈉、鋅、氧化鋅、硫酸鋅、及彼等之混合物。礦物質化合物之非限制性示例衍生物包括任何礦物質化合物之鹽、鹼鹽、酯和螯合物。
礦物質可以鹽之形式(諸如磷酸鈣、甘油磷酸鈣、檸檬酸鈉、氯化鉀、磷酸鉀、磷酸鎂、硫酸亞鐵、硫酸鋅、硫酸銅、硫酸錳及亞硒酸鈉)被添加在成長奶或其他兒童營養組成物中。其他維生素和礦物質可依本技藝所知者添加。
於一實施態樣中,每一份量之該兒童營養組成物中可含有的維生素A、C和E、鋅、鐵、碘、硒和膽鹼量為對任何指定國家之最大膳食建議量的約10至約50%,或對國家群組之平均膳食建議量的約10至約50%。於另一實施態樣中,每一份量之該兒童營養組成物可供應之維生素B量為對任何指定國家之最大膳食建議量的約10至30%,或對國家群組之平均膳食建議量的約10至30%。再於另一實施態樣中,該兒童營養產品中之維生素D、鈣、鎂、磷及鉀的數量可相當於乳品中所發現者之平均數量。於其他實施態樣中,每一份量之該兒童營養組成物中的其他營養素之存在量可為對任何指定國家的最大膳食建議量之約20%,或對國家群組之平均膳食建議量的約20%。
本發明之營養組成物可選擇擇地包括一或多
種以下調味劑,包括,但不限於調味之萃取物、揮發油、可可粉或巧克力調味劑、花生醬調味劑、餅乾碎片、香草或任何市售之調味劑。有用之調味劑的實例包括,但不限於純茴香萃取物、仿香蕉萃取物、仿櫻桃萃取物、巧克力萃取物、純檸檬萃取物、純柳橙萃取物、純薄荷萃取物、蜂蜜、仿鳳梨萃取物、仿蘭姆酒萃取物、仿草莓萃取物、葡萄或葡萄籽萃取物、蘋果萃取物、越橘萃取物或香草精;或揮發油,諸如香蜂花油(balm oil)、月桂油、佛手柑油、柏木油(cedarwood oil)、櫻花油、肉桂油、丁香油或薄荷油;花生醬、巧克力調味劑、香草餅乾碎片、奶油糖果、太妃糖及彼等之混合物。調味劑的量根據所使用之調味劑可有很大的變化。調味劑之類型和量可依本技藝所已知者選擇。
本發明之營養組成物可選擇地包括一或多種可添加在其中以用來穩定該最終產品之乳化劑。合適之乳化劑的實例包括,但不限於卵磷脂(例如來自卵、或大豆或任何其他植物及動物來源)、α-乳白蛋白及/或甘油單酯及甘油二酯,及彼等之混合物。熟習本技藝之人士可輕易明白其他乳化劑,而合適之乳化劑的選擇將部分取決於該調合物及最終產品。
本發明之營養組成物可選擇地包括一或多種亦可添加在其中以延長產品之貨架期的防腐劑。合適之防腐劑包括,但不限於山梨酸鉀、山梨酸鈉、苯甲酸鉀、苯甲酸鈉、乙二胺四醋酸二鈉鈣及彼等之混合物。
本發明之營養組成物可選擇地包括一或多種穩定劑。用於實行本發明之營養組成物的合適穩定劑包括,但不限於阿拉伯膠、印度樹膠(ghatti gum)、刺梧桐樹膠(karaya gum)、黃蓍膠、瓊脂、紅藻膠(furcellaran)、果阿膠(guar gum)、結冷膠、刺槐豆膠、果膠、低甲氧基果膠、明膠、微晶型纖維素、CMC(羧甲基纖維素鈉)、甲基纖維素、羥丙基甲基纖維素、羥丙基纖維素、DATEM(甘油單酯及甘油二酯之二乙醯基酒石酸酯)、右旋糖苷、角叉菜膠、CITREM及彼等之混合物。
於一些實施態樣中,本文所描述之營養組成物有利地減少個體之發炎反應。因此,本發明關於經由投予個體含有本文所描述之蛋白質等效來源之營養組成物來減少個體之促炎性反應的方法。例如,本方法可減少個體之促炎性細胞因子的製造。更特別地,本發明關於經由提供個體含有本文所描述之肽組分的營養組成物來減少製造IL-17的方法。
於一些實施態樣中,該用於減少個體之發炎反應的方法包含投予個體包含碳水化合物來源、蛋白質等效來源及脂肪來源之營養組成物,其中該蛋白質等效來源包括:a)包含SEQ ID NO 4、SEQ ID NO 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及SEQ ID NO 63之肽組份,及
b)完整蛋白質、水解蛋白質或彼等之組合。於一些實施態樣中,該肽組份可包含表1中所揭示之額外的肽類。例如,該組成物可包括至少10種表1中所揭示之額外的肽類。於一些實施態樣中,20%至80%之該蛋白質等效來源包含肽組份,且20%至80%之該蛋白質等效來源包含完整蛋白質、部分水解蛋白質及彼等之組合。
於另一實施態樣中,該方法包含投予個體一種營養組成物,其中20%至80%之該蛋白質等效來源包括一種肽組份,該肽組份包含至少3種選自由下列所組成之群組的肽類:SEQ ID NO 4、SEQ ID NO 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及SEQ ID NO 63,及至少5種選自表1之額外的肽類;且其中20%至80%之該蛋白質等效來源包含完整蛋白質、部分水解蛋白質或彼等之組合。
再於其他實施態樣中,該用於減少發炎反應之方法包括提供包含選自表1之肽組份的營養組成物,其中該肽組份係源自其莫耳質量分佈大於500道耳吞之酪蛋白水解產物。於一些實施態樣中,該酪蛋白水解產物之莫耳質量分佈係在500至2000道耳吞的範圍內。於其他實施態樣中,該用於減少發炎反應之方法包括提供包含本文所描述之肽組份的營養組成物,其中該肽組份係源自不包含莫耳質量分佈小於200道耳吞之肽類的酪蛋白水解產物。
於一些實施態樣中,該目標個體可為兒科個體。此外,於一實施態樣中,該提供予兒科個體之營養組成物可為嬰兒配方。本文中所鑑別並添加在嬰兒配方中之肽組份可選自特定來源且其濃度可經過調整以將健康益處最大化。於此方法之另一實施態樣中,該包含本發明之肽組份的營養組成物為成長奶。
於其中該營養組成物為嬰兒配方之實施態樣中,該組成物可有利地減少嬰兒體內之促炎性反應,從而減少發炎性疾病之發病率。此外,發炎性疾病減少可能會持續整個童年且持續到成年。類似地,當該營養組成物為成長奶時,攝入該成長奶之兒童可能會經歷成年時期及兒童時期發炎性疾病之發病率降低。
實施例1進一步說明用於製造蛋白質部分水解產物之方法。最初,將60.3公斤之非乳固體(奶粉)及37.4公斤之乳清蛋白濃縮物(60%)在含有54℃之水的槽中混合。該漿液之總固體含量係介於20%和23%之間。然後,測量該漿液之pH值。在該漿液中加入氫氧化鈉和氫氧化鉀,以將該漿液之pH值調節至7.0。調節pH值後,將0.5公斤之Amano N酶加入該漿液中。將Amano N酶加入該漿液後,使用氫氧化鈉和氫氧化鉀將pH值持續調節至pH 7.0。加入該漿液中之氫氧化鈉的總量為0.3公斤。加入該漿液中之氫氧化鉀的總量為1.5公
斤。
允許水解反應發生90分鐘,時間係從將Amano N酶加入漿液時開始計算。在90分鐘結束時,將漿液加熱處理以將該酶去活化。該熱處理包含將漿液之溫度提高至82℃持續10分鐘。在本實施例中取得之水解度係介於6%和9%之間。然後,將漿液冷卻並噴霧乾燥,以取得粉末狀水解產物。
實施例2提供測定該水解產物肽之分子量分佈的非限制性方法。
使用粒徑排阻層析法(SEC)來測定藉由目前描述之水解過程創建之水解產物肽的分子量分佈。具體地說,稱量足夠量之粉末狀嬰兒配方,以在50毫升錐形離心管中置入0.5克蛋白質。加入水以使管內之體積達到45毫升。將混合物置於Sarstedt D-5223混合器中並混合1小時。混合後,經由在管內加入另外5毫升的水以創建1%之蛋白質溶液。製備貯存標準物並混合1小時。
另外,將14.91克氯化鉀(KCl)加入1000毫升之燒杯中。在燒杯中加入700毫升水,以溶解該氯化鉀。然後,將250毫升乙腈及1.0毫升三氟醋酸加入該KCl溶液(洗提液)中。使用0.2M K2HPO4溶液將pH調整成3.0。
將HPCL試劑瓶填滿並以洗提液洗滌該瓶,
保留約50毫升以用於稀釋樣本及標準物。以洗提液淨化日立L-6200 A Intelligent Pump管線並以洗提液平衡該管柱一小時。
將樣本混合一小時後,將5.0毫升之各樣本移液入具螺旋蓋之玻璃試管中。亦將5.0毫升二氯甲烷移液到各個管中。將試管加蓋並藉由倒置4次來混合。然後,將該樣本在200×g離心5分鐘。
當樣本在離心機中時,以洗提液(800微升+3200微升)稀釋貯存標準物1-5。將約1毫升之各標準物移液到兩個自動進樣小瓶的各小瓶中並加蓋。
以洗提液(100微升+900微升)稀釋離心之樣本1-10的上(水)層。依下述將小瓶裝載入自動進樣器的托盤中:空白、標準物、樣本及第二標準物。將托盤置於日立自動進樣器中。使用在自動進樣器面板上之按鍵將欲測試之小瓶的全部數目輸入自動進樣器程式中,並測試樣本。結果指出該蛋白質之分子量的變化形廓。
實施例3係針對在原代人樹突細胞分析中藉由高度水解之酪蛋白及其大於500道耳吞之餾分來抑制促炎性細胞因子。具體的說,該被抑制之促炎性細胞因子包括介白素-12p70(“IL-12p70”)、干擾素-γ(IFN-γ)、介白素-8(“IL-8”)、腫瘤壞死因子-α(TNF-α)、介白素-6(“IL-6”)及介白素-1β(“IL-1β”)。
將分離出之CD14+單核細胞懸浮於RPMI培養基中且輔以10%經熱滅活之胎牛血清(“FBS”)、青黴素/鏈黴素、500IU/毫升之重組介白素-4(“rhIL-4”)及800IU/毫升之rhGM-CSF,並在37℃及5% CO2下,在組織培養瓶中培養5天,以允許分化成樹突細胞。
第5天,將細胞以20,000細胞/孔之密度接種於在96孔U底盤中的相同培養基中,再加入最終濃度為0.007、0.02、0.08及0.2%質量/體積之包含經高度水解的酪蛋白及其大於500道耳吞之餾分的水解產物樣本。使用最終濃度為1微克/毫升之地塞米松(“Dex”)作為用於抑制細胞因子製造的陽性對照組。所有樣本均以一式二份之生物複本測試。
第6天,以在不含GM-CSF及IL-4之培養基(RPMI+10%FBS+青黴素/鏈黴素)中的水解產物樣本更新細胞。加入水解產物一小時後,加入CD40L+增強劑(Alexis)使最終濃度為0.5微克/毫升。
然後,將細胞在37℃,5% CO2下培育24小時,接著收集上清液並儲存於-20℃直到根據製造商之指示,使用Meso Seale Discovery促炎性細胞因子多重分析來定量細胞因子。
如第1-2圖中所示,高度水解之酪蛋白及該高度水解之酪蛋白之大於500道耳吞的餾分可抑制經活化之原代人樹突細胞分泌促炎性細胞因子介白素-12p70(IL-12p70)。
如第3-4圖中所示,高度水解之酪蛋白及該高度水解之酪蛋白之大於500道耳吞的餾分可抑制經活化之原代人樹突細胞分泌促炎性細胞因子干擾素-γ(IFN-γ)。
如第5-6圖中所示,高度水解之酪蛋白及該高度水解之酪蛋白之大於500道耳吞的餾分可抑制經活化之原代人樹突細胞分泌促炎性細胞因子介白素-8(IL-8)。
如第7-8圖中所示,高度水解之酪蛋白及該高度水解之酪蛋白之大於500道耳吞的餾分可抑制經活化之原代人樹突細胞分泌促炎性細胞因子腫瘤壞死因子-α(TNF-α)。
如第9-10圖中所示,高度水解之酪蛋白及該高度水解之酪蛋白之大於500道耳吞的餾分可抑制經活化之原代人樹突細胞分泌促炎性細胞因子介白素-6(IL-6)。
如第11-12圖中所示,高度水解之酪蛋白及該高度水解之酪蛋白之大於500道耳吞的餾分可抑制經活化之原代人樹突細胞分泌促炎性細胞因子介白素-1β(IL-1β)。
實施例4係針對在原代人巨噬細胞分析中藉由高度水解之酪蛋白及其大於500道耳吞之餾分來抑制促
炎性細胞因子腫瘤壞死因子-α(“TNF-α”)。
將分離出之單核細胞懸浮於輔以10%經熱滅活之胎牛血清(FBS)及青黴素/鏈黴素的RPMI培養基中。加入重組人M-CSF,使最終濃度為100毫微克/毫升,並將細胞在37℃及5% CO2下培育5天。
第5天,將最終濃度為0.007、0.02、0.08及0.2%質量/體積之包含高度水解之酪蛋白及其大於500道耳吞之餾分的水解產物樣本加入帶有M-CSF之RPMI培養基中。
第6天,移除包含該水解產物樣本之培養基並以在不含M-CSF之培養基中的水解產物樣本更新細胞。加入水解產物一小時後,加入LPS(大腸桿菌(0111:84)),使最終濃度成為10毫微克/毫升。然後,將細胞在37℃和5% CO2下培育24小時,接著收集上清液並儲存於-20℃下,直到根據製造商之指示,使用Meso Scale Discovery TNF-α分析來定量TNF-α。
如第13-14圖中所示,高度水解之酪蛋白及該高度水解之酪蛋白之大於500道耳吞的餾分可抑制經活化之原代人巨噬細胞分泌促炎性細胞因子腫瘤壞死因子-α(TNF-α)。
實施例5係針對在非肥胖型糖尿病(NOD)小鼠之迴腸中評估促炎性細胞因子傳信者RNA的表現。
簡單的說,使用年齡相符之雌性BALB/C小鼠作為健康對照組小鼠。若未另外指明,在開始斷奶時使BALB/C小鼠維持例常食物,而NOD小鼠則維持例常食物或含高度水解之酪蛋白(CH)的膳食調合物。
藉由RNA分離套組將RNA從迴腸樣本分離出。合成cDNA並藉由定量性實時PCR估計細胞因子之表現。經由運行標準物並以甘油醛-3-磷酸脫氫酶(GAPDH)作為裝載對照組來計算RNA之絕對複本數(分子/微克)。
從第15-18圖中可察知,與年齡相符之健康(BALB/C)對照小鼠相比,NOD小鼠之迴腸中促炎性細胞因子IL-1、IL-6、IL-18及IL-17的分泌增加。經餵食高度水解之酪蛋白的飲食之NOD小鼠中的這些增加之數量減少。
實施例6為針對從非肥胖型糖尿病(NOD)小鼠之腸引流淋巴結中分離出之T細胞所分泌之促炎性細胞因子的變化形廓之評估。
簡單的說,使用年齡相符之雌性BALB/C小鼠作為健康對照組小鼠。若未另外指明,在開始斷奶時使BALB/C小鼠維持例常食物,而NOD小鼠則維持例常食物或含高度水解之酪蛋白(CH)的膳食調合物。
以在完整培養基(帶有10%(體積/體積)
FBS、L-麩醯胺2毫莫耳/升、100U/毫升青黴素加鏈黴素之DMEM)中之與培養板結合的抗CD3刺激來自各組小鼠之經分離出的腸繫膜淋巴結細胞。48小時後收集上清液餾份,根據製造商之說明以多重細胞因子珠陣列進行細胞因子分析。
如第19-21圖中所示,與年齡相符之健康(BALB/C)對照組小鼠相比,來自NOD小鼠之腸引流淋巴結T細胞的促炎性細胞因子IFN-γ、IL-4及IL-17的分泌增加。藉由高度水解之酪蛋白的飲食,NOD小鼠中的這些增加之數量減少。
實施例7係針對小鼠巨噬細胞株中細胞因子分泌的分析。
將J774A.1小鼠巨噬細胞株維持在輔以10%(體積/體積)胎牛血清、50IU/毫升青黴素及0.5毫克/毫升鏈黴素之RPMI-1640中。將細胞在37℃及5% CO2下培育。
將濃度為100毫克/毫升且通過0.2微米尼龍過濾器過濾滅菌之酪蛋白水解產物樣本加入無菌水中,該酪蛋白水解產物樣本包括高度水解之酪蛋白及其大於500道耳吞之餾分。亦測試未經過濾滅菌之高度水解的酪蛋白以作為額外的對照組。將細胞以1×106細胞/毫升之密度平皿接種在盤中,與最終濃度為1毫克/毫升之水解產物一
起培育1小時,一式三份。然後,或者以100毫微克/毫升LPS處理細胞,或者不處理細胞24小時,再藉由離心收集上清液並儲存在-20℃下。
根據製造商之指示,使用用於IL-1β、IL-6、IL-12p40及TNF-α之個別ELISA分析來評估上清液中之各細胞因子的濃度。以一式三份測試樣本,並在graphpad prism 6軟體上使用平常之單因子ANOVA計算差異之顯著性。p<0.05被認為具顯著性差異。
如第22-25圖中可察知,藉由高度水解之酪蛋白(包括在細胞培育前有或無進行無菌過濾者)可減少從以LPS刺激之小鼠巨噬細胞分泌促炎性細胞因子IL-12p40及IL-1β(*p<0.05)。高度水解之酪蛋白顯著減少促炎性細胞因子IL-6之分泌(*p<0.05)。該高度水解之酪蛋白的大於500道耳吞之餾分顯著減少促炎性細胞因子TNF-α之分泌(*p<0.05)。結果亦顯示在第26圖之表格格式中。
表3提供包括來自表1之8種肽的肽組份之示例性實施例。
表4提供包括來自表1之某些肽類的肽組份之示例性實施例。
表5提供本發明之營養組成物的示例性實施態樣並描述每100千卡之份量中所欲包含之各成分的量。
本專利說明書中所引用之所有參考資料,包括,但不限於所有論文、出版物、專利案、專利申請案、演示、文本、報告、手稿、手冊、書籍、網路貼文、雜誌文章、期刊,等之全部內容在此以引用方式納入本專利說
明書中。本文中參考資料的討論只是為了概述由其作者所下之斷言,並非承認任何引用構成先前技藝。本申請人保留質疑所引用之參考資科的準確性和相關性之權利。
雖然本揭露內容之實施態樣已使用特定術語、裝置和方法描述,這類描述僅用於說明。所使用之文字是描述,而非限制之文字。可理解的是,本技藝之一般技術人士可在不背離本揭露內容之精神和範圍(這些列於接續之申請專利範圍中)的情況下作出各種改變和變化。此外,應理解的是,各種實施態樣之觀點可以全部或部分互換。例如,雖然已示例用於生產根據那些方法製造之市售無菌液體營養補充劑的方法,仍可以考慮其他用途。因此,所附之申請專利範圍的精神和範圍不應只限於其中所包含之版本的說明。
<110> 營養股份有限公司 亞太
<120> 包含具有抗發炎性質之肽組份之營養組成物及其用途
<130> MJE00375NP
<160> 64
<170> PatentIn version 3.5
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Claims (20)
- 一種營養組成物,其包含:(i)碳水化合物來源;(ii)脂肪來源;及(iii)蛋白質等效來源,其中a)20%至80%之該蛋白質等效來源包括含有SEQ ID NO 4、SEQ ID NO 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及SEQ ID NO 63之肽組份;且b)20%至80%之該蛋白質等效來源包含完整蛋白質、部分水解蛋白質或彼等之組合。
- 如申請專利範圍第1項之營養組成物,其中該肽組份之存在量為約0.2克/100千卡至約5.6克/100千卡。
- 如申請專利範圍第1項之營養組成物,其中該肽組份進一步包含至少10種選自表1之額外的肽類。
- 如申請專利範圍第1項之營養組成物,其中該蛋白質等效來源包含水解度小於40%之部分水解蛋白質。
- 如申請專利範圍第1項之營養組成物,其進一步包含至少一種長鏈多元不飽和脂肪酸。
- 如申請專利範圍第5項之營養組成物,其中該至少一種長鏈多元不飽和脂肪酸係選自由下列所組成之群組:二十二碳六烯酸及花生四烯酸。
- 如申請專利範圍第1項之營養組成物,其進一步包含來自益生菌分批培養過程之指數生長末期的培養上清液,以用於治療或預防由病原體造成之感染。
- 如申請專利範圍第1項之營養組成物,其進一步包含益生菌。
- 如申請專利範圍第1項之營養組成物,其進一步包含益菌素。
- 如申請專利範圍第1項之營養組成物,其中當被投予個體時,該營養組成物減少製造選自由下列所組成之群組的促炎性細胞因子:IL-12、IL-17、IL-5、IL-4、IFN-γ、IL-8、TNF-α、IL-6及IL-1。
- 一種營養組成物,其包含:(i)碳水化合物來源;(ii)脂肪來源;及(iii)蛋白質等效來源;其中a)20%至80%之該蛋白質等效來源包括肽組份,且該肽組份含有至少5種選自表1之肽類及至少3種選自表2之額外的肽類;且b)20%至80%之該蛋白質等效來源包含完整蛋白質、部分水解蛋白質或彼等之組合。
- 如申請專利範圍第11項之營養組成物,其包含至少10種選自表1之額外的肽類。
- 如申請專利範圍第11項之營養組成物,其中該蛋 白質等效來源包含莫耳質量分佈大於500道耳吞之酪蛋白水解產物。
- 如申請專利範圍第11項之營養組成物,其中該蛋白質等效來源包含水解度小於40%之部分水解蛋白質。
- 如申請專利範圍第11項之營養組成物,其進一步包含益菌素。
- 如申請專利範圍第11項之營養組成物,其進一步包含來自益生菌分批培養過程之指數生長末期的培養上清液,以用於治療或預防由病原體造成之感染。
- 一種用於減少個體的發炎反應之方法,其包含提供該個體包含下列之營養組成物:碳水化合物來源,脂肪來源,及蛋白質等效來源,其中20%至80%之該蛋白質等效來源包括含有SEQ ID NO 4、SEQ ID NO 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及SEQ ID NO 63之肽組份,且其中20%至80%之該蛋白質等效來源包含完整蛋白質、部分水解蛋白質或彼等之組合。
- 如申請專利範圍第17項之方法,其中該肽組份進一步包含至少10種選自表1之額外的肽類。
- 如申請專利範圍第17項之方法,其中該蛋白質等 效來源包含水解度小於40%之部分水解蛋白質。
- 如申請專利範圍第17項之方法,其中該營養組成物減少製造選自由下列所組成之群組的促炎性細胞因子:IL-12、IL-17、IL-5、IL-4、IFN-γ、IL-8、TNF-α、IL-6及IL-1。
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2013
- 2013-03-15 US US13/833,094 patent/US8889633B2/en active Active
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2014
- 2014-03-11 CN CN201480015753.8A patent/CN105072928B/zh active Active
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- 2014-03-11 WO PCT/US2014/023608 patent/WO2014150558A1/en active Application Filing
- 2014-03-11 MY MYPI2015702455A patent/MY172218A/en unknown
- 2014-03-11 SG SG11201505689PA patent/SG11201505689PA/en unknown
- 2014-03-11 PE PE2015001979A patent/PE20151978A1/es not_active Application Discontinuation
- 2014-03-11 MX MX2015011934A patent/MX365415B/es active IP Right Grant
- 2014-03-11 AU AU2014237014A patent/AU2014237014B2/en active Active
- 2014-03-11 CA CA2905799A patent/CA2905799C/en not_active Expired - Fee Related
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- 2014-03-11 EP EP14717284.5A patent/EP2983531B1/en active Active
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US20150093463A1 (en) | 2015-04-02 |
PE20151978A1 (es) | 2016-01-23 |
MY172218A (en) | 2019-11-18 |
ES2736313T3 (es) | 2019-12-27 |
CA2905799A1 (en) | 2014-09-25 |
MX2015011934A (es) | 2015-12-01 |
HK1216976A1 (zh) | 2016-12-16 |
MX365415B (es) | 2019-06-03 |
CN105072928A (zh) | 2015-11-18 |
US8889633B2 (en) | 2014-11-18 |
AU2014237014A1 (en) | 2015-08-13 |
EP2983531A1 (en) | 2016-02-17 |
AR095332A1 (es) | 2015-10-07 |
RU2015140269A (ru) | 2017-04-21 |
CA2905799C (en) | 2021-03-30 |
NZ710020A (en) | 2020-04-24 |
WO2014150558A1 (en) | 2014-09-25 |
BR112015018335A2 (pt) | 2017-08-22 |
PH12015502071A1 (en) | 2016-01-25 |
US9457058B2 (en) | 2016-10-04 |
EP2983531B1 (en) | 2019-04-24 |
US20140271553A1 (en) | 2014-09-18 |
CN105072928B (zh) | 2021-02-12 |
PH12015502071B1 (en) | 2016-01-25 |
SG11201505689PA (en) | 2015-08-28 |
AU2014237014B2 (en) | 2018-02-22 |
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