CN105705037A - 用于抑制肿瘤生长的营养组合物 - Google Patents
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- CN105705037A CN105705037A CN201380080014.2A CN201380080014A CN105705037A CN 105705037 A CN105705037 A CN 105705037A CN 201380080014 A CN201380080014 A CN 201380080014A CN 105705037 A CN105705037 A CN 105705037A
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Abstract
用于抑制肿瘤生长的营养组合物,包含:作为蛋白质的乳蛋白水解物和发酵乳蛋白;作为脂质的含油酸的油和乳磷脂和/或大豆卵磷脂;作为碳水化合物的异麦芽酮糖。作为口服或管饲营养物,所述组合物可用于抑制肿瘤(癌症)患者的肿瘤生长。
Description
技术领域
本发明涉及营养组合物,所述营养组合物可用于肿瘤患者的营养管理和治疗。此外,本发明涉及可用于病理性改善结肠实体肿瘤患者的营养组合物。
背景技术
近几年,报道了使用营养组合物来治疗肿瘤患者。在申请号为CN20061054681的中国专利申请中,报道了豆浆、蔬菜汁和鸡蛋营养米粉以及小麦面粉食品。
含有大豆异黄酮成分的豆浆能有效地抑制白血病、结肠癌、肺癌、胃癌等肿瘤的发生;胡萝卜汁中的黄酮和丰富的抗氧化成分,对人体有很强的抗癌效果,鸡蛋中含有丰富的蛋白质给人体补充营养,提高人体免疫力,预防疾病的发生。
在申请号为CN20081006602的另外一件中国专利申请中,描述了含有短肽和益生菌的特殊营养食品配方。该食品配方使用来源于食物的短肽作为原料,添加了益生菌和功能性低聚糖形成复合配方。以该配方,可制备干粉食品,并用冷水冲服。
根据该配方制备的食品可提高生物体蛋白质的营养状况,优化肠道的微生态环境,调节肠道的功能,预防便秘、腹泻和结肠癌,提高机体免疫力,美白,全面增强体质。
在公开号为WO2004002238的国际专利申请中,报道了一种营养组合物,该组合物结合了益生元源和能够诱导哺乳动物酶活性的植物化学成分两者的附加效果。提供了能降低患癌症风险的营养组合物。
在公开号为WO2007107295的国际专利申请中,提供了一种包含异麦芽酮糖,白菌二糖,海藻酮糖(trehalulose)和/或松二糖的低血糖食品组合物,作为低血糖碳水化合物来源。低血糖食品用于治疗和/或预防人体或者动物体的恶性肿瘤疾病。
在公开号为JP10-203994的日本专利申请中,提供了一种组合物,该组合物包含作为活性成分的500-5000分子量的肽,该肽通过使蛋白酶作用于至少一种蛋白质而形成,该蛋白质选自:源于牛奶、大豆和酵母的蛋白质。该组合物在免疫增强,抗应激,抗氧化性,降血压,抗癌,降低中性脂肪(neutralfatdepressive)和超高密度脂蛋白胆固醇活性方面表现优异。
以上是一些涉及有关治疗肿瘤的营养组合物的专利以及专利申请文件,但是这一领域还没有这样的液体膳食型营养组合物,该组合物含有预定能量百分比的蛋白质、脂质和碳水化合物,并添加了粘性的可溶性食物纤维和菊粉或水解物。
现有技术文件
专利文献
PLT1:CN20061054681
PLT2:CN20081006602
PLT3:WO2004002238A1
PLT4:WO2007107295A1
PLT5:JP10-203994
PLT6:JP2001-181303
PLT7:WO01049308
发明内容
本发明要解决的问题
本发明的目标是提供抑制肿瘤生长的营养组合物,特别是抑制结肠肿瘤的生长。
解决问题的方法
本发明的发明人提供可抑制肿瘤生长的营养组合物,如减小肿瘤重量和体积,且具有光滑边缘和在病理组织学检查中密集生长的细胞。
本发明的有益效果
本发明提供了一种用于肿瘤患者营养管理和治疗的营养组合物。例如,摄入的患者通过服用流体膳食(fluiddiet)、肠内营养、经口营养、输卵管供给而能够抑制肿瘤生长。进一步,本发明更适用于营养管理安全和康复护理期间。
本发明的营养组合物可以抑制肿瘤生长,同时提供营养。营养组合物可被用于,例如,液体食物,经口/肠内/管供给营养物来抑制摄入该组合物的患者的肿瘤生长。而且,本发明的组合物可安全地摄入,因此适合长期治疗的营养管理。
附图说明
图1是S–180荷瘤小鼠的存活曲线。
图2是C26荷瘤小鼠的食物摄入图。
图3是C26荷瘤小鼠的体重的曲线图。
图4是从C26荷瘤小鼠解剖的肿瘤照片。
图5是C26荷瘤小鼠的肿瘤体积的曲线图。
图6是C26肿瘤组织(苏木精-伊红染色,×100)的病理学照片。
具体实施方式
本发明将在以下被详细描述。
1.蛋白质
1-1.乳蛋白水解物
酪蛋白、乳清蛋白(乳清蛋白浓缩物(WPC)、乳清蛋白分离物(WPI)、α-乳白蛋白(α-La)、和β-乳球蛋白(β-Lg))、乳蛋白浓缩物(MPC或全乳蛋白(TMP)),这些都可作为蛋白质来源。
通常用于水解乳清蛋白的酶为,例如,胃蛋白酶、胰蛋白酶、和胰凝乳蛋白酶。然而,也有研究报道使用植物来源的木瓜蛋白酶,以及来自细菌和真菌的蛋白酶(FoodTechnol.,48:68-71,1994;TrendsFoodSci.Technol.,7:120-125,1996;FoodProteinsandTheirApplications:pp.443-472,1997)。乳清蛋白水解酶活性变化很大。胃蛋白酶降解已变性的α-乳白蛋白和α-乳白蛋白,但是不能降解天然的β-乳球蛋白(Neth.MilkdairyJ.,47:15-22,1993)。胰蛋白酶可缓慢水解α-乳白蛋白,但是几乎不能降解β-乳球蛋白(Neth.MilkdairyJ.,45:225-240,1991)。
胰凝乳蛋白酶可迅速降解α-乳白蛋白,然而,却缓慢降解β-乳球蛋白。
木瓜蛋白酶可水解牛血清白蛋白(BSA)和β-乳球蛋白,但是对α-乳白蛋白显示抗性(Int.DairyJournal6:13-31,1996a)。然而,在酸性pH值下,不与Ca键合的α-乳白蛋白可被木瓜蛋白酶完全降解(J.DairySci.,76:311-320,1993)。
通过控制乳蛋白的酶促降解和修饰蛋白质,在宽的pH范围和加工条件下可以改变该蛋白质的功能特性(EnzymeandChemicalModificationofproteinsinFoodproteinsandtheirApplicationspp.393-423,1997MarcelDekker,Inc.,NewYork,1997;FoodTechnol.,48:68-71,1994)。
肽键的水解增加了带电基团的数目和疏水性,降低了分子量,并且修改了分子构型(J.DairySci.,76:311-320,1993)。功能性质的变化很大程度上取决于水解的程度。
例如,通常观察到的乳清蛋白功能的最大的变化是溶解度增加和粘度下降。通常当水解度高时,即使加热,水解物也不会沉淀,并且在pH3.5到4.0下高度可溶。水解物的粘度也远远低于完整的蛋白质。这些差异在蛋白质浓度高时尤为突出。其他影响包括改变的凝胶的性质、提高的耐热性、增加的乳化和起泡能力、降低的乳液和泡沫稳定性(Int.Dairyjournal,6:13-31,1996a;DairyChemistry4,pp.347-376,1989;J.DairySci.,79:782-790,1996)。
已知各种源于乳蛋白的生物活性寡肽。例如,包含经处理的原料的癌症转移抑制剂,该原料是通过培养蛋白质消化酶(如,胃蛋白酶)得到的(JP62059220(A))。
例如,已知蛋白质消化酶(如胃蛋白酶)的自溶产物,通过肿瘤细胞的血小板聚集诱导活性来抑制癌症转移(JPA62-059220)。
已知源于乳蛋白的各种生理活性肽。例如,已知的蛋白质消化酶(如胃蛋白酶)的自溶产物,通过肿瘤细胞的血小板聚集诱导活性来抑制癌症转移。进一步,还已知的是在抗体疗法中,乳铁蛋白水解物增强了抗体制剂的细胞毒活性。
除了以上引用的参考文献,存在许多有关乳蛋白水解物的专利(公开的专利申请和专利)。实例包括:专利之一,分别水解酪蛋白和乳清蛋白,然后吸附和去除疏水部分,接着将酪蛋白和乳清蛋白以指定的比例混合(日本专利No.2,986,764);专利之二,用源自芽孢杆菌和放线菌的蛋白酶水解乳清蛋白,然后去除所述酶和不溶性水解产品的专利(日本专利No.3,222,638);专利之三,其涉及肽混合物,在所述肽混合物中,通过酶促降解β-乳球蛋白获得的支链氨基酸/芳香族氨基酸的摩尔比为10重量%或更多,其中芳香族氨基酸小于2.0重量%,并且其中平均分子量为几百到几千(日本专利No.3,183,945);专利之四,在乳清蛋白中选择性酶促降解β-乳球蛋白(日本专利No.2,794,305);和专利之五,使用源自地衣芽孢杆菌和/或枯草杆菌的蛋白酶,通过非pH值稳态(non-pH-stat)技术将乳清蛋白水解至15%-30%(葡萄糖当量;DE),然后从截留分子量大于10,000的超滤膜获得透过液(日本专利No.3,167,723);且除了这些专利和专利申请,本发明包括其他专利和未审的公开专利。
乳清蛋白的蛋白水解物可通过以下五个步骤制备。下面的步骤(1)到(5)描述了乳清蛋白水解物的制备方法。
(1)将作为干燥产品的、蛋白质含量约为90%(w/w)的乳清蛋白分离物(WPI,戴维斯克国际食品有限公司),溶解在蒸馏水中,以提供8%(w/v)的蛋白质浓度,从而得到蛋白质水性溶液。
(2)将所述水性溶液在85℃下加热2min使蛋白质变性。热处理的水性溶液的pH约为7.5。
(3)然后,加入2.4L相对于蛋白质(底物)的浓度为2.0%(w/w)的碱性蛋白酶(酶,诺维信),将水性溶液保持在55℃下进行水解3小时。
(4)随后,加入相对于蛋白质(底物)浓度为3.0%(w/w)的源于猪的胰蛋白酶PTN6.0S(酶,诺维信日本有限公司),将水性溶液保持在55℃下进行水解3小时。换句话说,总的水解时间为6小时。完成这些水解反应后,水性溶液的pH大约在7.0。
(5)将乳清蛋白水解物离心(20,000×g,10分钟),使用截留分子量为10,000的超滤膜处理(MilliporeCorporation,UltrafreeMC)。
在制备蛋白质水解物的方法中,关于蛋白质水解的制备方法,例如,选择预热、酶底物的比例(E/S)、pH、水解温度以及水解时间作为五个参数进行优化。预热:65-90℃,E/S:0.01-0.2,水解温度:30-65℃,水解时间:3小时至小于20小时。
用于制备蛋白水解物的酶的实例包括以下来自诺和诺德的酶:
(1)内切蛋白酶
源于地衣芽孢杆菌:碱性蛋白酶
源于缓慢芽孢杆菌(B.lentus):益瑞蛋白酶
源自枯草芽孢杆菌:中性蛋白酶(Neutrase)
源于细菌:复合蛋白酶
源于猪胰脏:PTN(胰蛋白酶)
(2)外切蛋白酶
源于米曲霉:风味蛋白酶
源于猪或牛内脏:羧肽酶
除了上述酶的例子,包括源于动物的胰酶、胃蛋白酶、源于植物的木瓜蛋白酶、菠萝蛋白酶、源于微生物的内切蛋白酶和外切蛋白酶(例如,乳酸菌、酵母菌、霉菌和分枝杆菌)、它们未精制的纯化原料和细菌匀浆。此外,当组合用酶时,源于地衣芽孢杆菌的碱性蛋白酶和源于猪胰脏的PTN(胰蛋白酶)的组合经常被使用。
本发明的蛋白水解物包括:酶水解物,超滤后的保留液或透过液,和表现相似活性的市售乳蛋白水解物。例如,本发明的蛋白水解物可以使用截留分子量为10,000的超滤膜的保留液。例如,截留分子量为10,000的超滤膜的保留液可用作本发明的蛋白水解物。
乳蛋白的结合率可以通过添加乳蛋白水解物的量进行调节,乳蛋白结合率可依据加入的其他成分(发酵乳蛋白、含油酸的脂肪和油、乳磷脂、大豆卵磷脂、异麦芽酮糖或类似物),癌症病人的病理、症状、年龄、体重,使用目的或类似情况,进行适当调整。具体地,添加乳蛋白水解物的量的实例包括每100mL营养组合物0.9-5.0g,优选0.9-3.0g,更优选1.0-2.5g,更优选1.2-2.0g,但不限于这些范围。
1-2发酵的乳蛋白
发酵乳(使用发酵剂(starter)如乳酸菌通过发酵液态奶得到的任何产品,所述液态奶通过混合一种或多种家畜奶,如牛奶、水牛奶、山羊奶、绵羊奶、或马奶和/或乳原料,如部分脱脂乳、脱脂乳、还原全脂乳(reducedwholemilk)、还原脱脂乳、还原部分脱脂乳、黄油、它们的奶油或类似物而制备)可作为本发明中发酵乳蛋白的来源。例如,新鲜奶酪、天然奶酪、酸奶和乳清干酪包含在本发明的发酵乳中。另外,本发明的奶酪,指的是那些通过使用乳酸菌对乳、酪乳或奶油进行发酵而获得的那些,或通过加入酶至乳、酪乳或奶油而获得的凝乳,其中乳清被去除,且凝固或陈化的存在是无关紧要的。通常使用保加利亚乳杆菌和嗜热链球菌作为生产发酵乳的发酵剂,但不局限于此,且实例包括乳酸杆菌和双歧杆菌,如乳酸链球菌、乳脂链球菌、醋酸乳酸双重链球菌、屎肠球菌(Enterococcusfaecium)、粪肠球菌(Enterococcusfecalis)、干酪乳杆菌、瑞士乳杆菌、嗜酸乳杆菌、鼠李糖乳杆菌(Lactobacillusrhamnosis)、植物乳杆菌、鼠乳杆菌(Lactobacillusmurinus)、罗伊氏乳杆菌、短乳杆菌、加氏乳杆菌、长双歧杆菌、两岐双岐杆菌以及短双岐杆菌。此外,所使用的微生物,如丙酸菌属,在生产发酵乳时,可组合使用。本发明的营养组合物可以用任何发酵乳制成,但优选使用新鲜奶酪或酸奶制备,更优选使用夸克奶酪(quark)或酸奶制备。
新鲜奶酪有很多种,如乡村奶酪、夸克奶酪、纤丝奶酪、纽沙尔特干酪、奶油奶酪、马苏里拉奶酪、意大利乳清奶酪以及马斯卡彭奶酪。夸克奶酪是一种未陈化(新鲜的)的奶酪,脂肪含量低,具有新鲜风味和酸味的特点。生产夸克奶酪的典型方法描述如下。先对脱脂乳巴氏杀菌,然后接种0.5-5%的发酵剂乳酸杆菌(主要是保加利亚乳杆菌和嗜热链球菌)进行发酵。当溶液的pH值达到4.6的时候,形成凝乳,将所获得的凝乳冷却后用夸克奶酪分离器将乳清离心分离。本发明的夸克组合物的实例为17-19%(w/w)的总固体含量,11-13%(w/w)的蛋白质,低于1%(w/w)的脂肪,2-8%(w/w)的碳水化合物和低于2%(w/w)的乳糖。此外,使用凝乳酶凝固的奶酪也包括在本发明的夸克奶酪中。进一步,那些通过向脱脂奶中加入乳球菌属的乳酸菌和乳脂菌(cremoris)和属于明串珠菌属菌种的混合培养液所培养的、并去除了乳清的奶酪,也包括在本发明的夸克奶酪中。更进一步,那些通过使用刀具切割以与上面的方法同样的方式获得的凝乳和加热发酵液分离乳清的奶酪,也包括在本发明的新鲜奶酪中。
已知乳酸菌和其培养物上清液具有免疫调节功能。例如,已知乳酸乳球菌乳脂亚种(Lactococcuslactissubsp.cremoris)和源于其培养物上清液的多糖可选择性地对转化细胞(肿瘤、癌细胞等)有毒性(日本专利,公开号2009-256312)。
根据其他组分(乳蛋白水解物、含油酸的脂肪和油、乳磷脂、大豆卵磷脂、异麦芽酮糖或类似物)的添加量、摄取对象的病理、症状、年龄、体重、以及使用目的等,适当调节发酵乳蛋白的添加量。具体地,发酵乳蛋白添加量的实例包括,0.5-6g,优选2-6g,更优选2.5-4.5g的蛋白质每100mL营养组合物,但不限于这些范围。
日本部级条例(卫生福利部第52号条例,1951年12月27日)就奶和奶制品涉及的成分标准,建立了“禁止从分娩5天内的牛、山羊或绵羊挤奶”的规定,作为乳等通用的成分标准和制备方法的准则。换句话说,该部级条例限制了初乳在乳制品中的使用。
在本发明中,优选地,用于制备乳蛋白和发酵乳蛋白的水解物的乳是普通乳(或者被称为成熟乳)。乳可能源自任何动物,如奶牛、水牛、山羊、绵羊、马、或人。
2.脂质
2-1.磷脂
乳磷脂和源于大豆的卵磷脂或蛋黄卵磷脂的结合作为磷脂使用。乳磷脂也可单独使用。在生物化学、医学和药理学领域,术语“卵磷脂”仅指代磷脂酰胆碱。然而在商业或工业领域,卵磷脂作为磷脂酰胆碱、磷脂酰乙醇胺、磷脂酰肌醇、磷脂酸和其它磷脂混合物的通用术语使用。在“日本食品添加剂的规范和标准(Japan'sSpecificationsandStandardsforFoodAdditives)”中,第七版(1999),卵磷脂被定义为“从油籽或动物源获得的物质,其主要成分是磷脂”。
乳磷脂包含鞘磷脂(sphingoyelin)(SM)、磷脂酰胆碱(PC)、磷脂酰乙醇胺(PE)、磷脂酰肌醇(PI)、磷脂酰丝氨酸(PS)和溶血磷脂酰胆碱(LPC),并且仅存在于乳脂球膜(MFGM)中。表1是MFGM磷脂级分的组成(BulletinofJapanDairyTechnicalAssociation,第50卷,第58-91页,2000)。
如表1所示,乳磷脂的特征是含有大量的鞘磷脂,所述鞘磷脂没有包含在大豆卵磷脂内。
[表1]
磷脂成分 | 重量% |
鞘磷脂 | 22 |
磷脂酰胆碱(Shosphatidylcholine) | 36 |
磷脂酰乙醇胺 | 27 |
磷脂酰肌醇 | 11 |
磷脂酰丝氨酸 | 4 |
溶血磷脂酰胆碱 | 2 |
大豆卵磷脂
虽然大豆卵磷脂作为天然食品添加剂,被广泛地用于食品和食品产品领域,但多烯磷脂酰胆碱也被用作药物(用途:在慢性肝脏疾病中改善肝功能、脂肪肝和高血脂)。
通常根据磷脂酰胆碱含量对“天然的”卵磷脂产品进行归类。生产了根据它们的用途改善的各种类型的卵磷脂。如表2所示,基于纯化和大豆卵磷脂分级,根据主要成分磷脂酰胆碱的差别,对大豆卵磷脂产品进行了适当的分类(Fujikawa,T.,OilChemistry,Vol.40(10):951-958,1991)。
[表2]
表2
类型 | 磷脂酰胆碱含量(%) |
糊状卵磷脂(Paste lecithin) | 15-20 |
纯化卵磷脂 | 20-25 |
提取卵磷脂 | 30-40 |
磷脂酰胆碱浓缩卵磷脂 | 45-60 |
磷脂酰胆碱高度纯化卵磷脂 | 75-95 |
磷脂(PC,PE,PS,PG,等) | 分别为98%或更多 |
在本发明中,乳磷脂和大豆卵磷脂可单独或组合使用。加入的乳磷脂和/或大豆卵磷脂的量可依据其他成分(乳蛋白水解物、发酵乳蛋白、含油酸的脂肪和油、异麦芽酮糖等)的添加量、摄入对象的病理、病症、年龄、体重、以及使用目的进行适当调整。具体地,乳磷脂和/或大豆卵磷脂添加量的实例包括0.01-0.5g,0.05-0.5g,0.1-0.5g和0.2-0.3g的总量每100mL营养组合物,但不限于这些范围。
2-2.其他脂质
健康、劳动和福利部建议饱和脂肪酸(SFA:棕榈酸、硬脂酸等):单不饱和脂肪酸(MUFA:油酸等):多不饱和脂肪酸(PUFA:亚油酸、亚麻酸等)的优选摄入比例从之前的1:1.5:1变为3:4:3,且n-6脂肪酸:n-3脂肪酸比例为4:1.这个建议的原因之一是,在日本很难实行单不饱和脂肪酸摄入比例是饱和脂肪酸和多不饱和脂肪酸的1.5倍的饮食。因此,在脂质的脂肪酸组成中,单不饱和脂肪酸含量提高。将为单不饱和脂肪酸的油酸,混合入脂肪酸组成中,构成超过30%、或优选30-60%的混合物。含有大量的油酸的脂质来源,包括高油酸葵花籽油、油菜籽油、橄榄油、高油酸红花油、大豆油、玉米油和棕榈油。更进一步,营养调和的油和脂肪(NOF公司)也是含油酸脂质来源。也可使用葵花籽油、油菜籽油、橄榄油和含橄榄油的混合物。
油酸含量可随着其他成分(如源于发酵乳的蛋白质)含量进行调整。乳卵磷脂和大豆卵磷脂可单独或组合使用。每100mL产品的总含量可以是0.1-0.5g,或优选0.2-0.3g。油酸含量可以是2-3g,或优选2.1-2.5g。更进一步,加入多不饱和脂肪酸,如DHA、EPA和花生四烯酸,和中链脂肪酸,如辛酸、癸酸和月桂酸,来调节饱和脂肪酸:单不饱和脂肪酸:多不饱和脂肪酸的比例至3:4:3。
加入油酸的量可根据其他成分(乳蛋白水解物、发酵乳蛋白、乳磷脂、大豆卵磷脂、异麦芽酮糖等)的添加量、摄入对象的病理、病症、年龄、体重、以及使用目的或类似情况进行适当调整。具体地,在本发明营养组合物的脂肪酸组成中,油酸加入量实例包括25%或更多,优选30%或更多,且更优选30-50%,但不限于这些范围。进一步,加入多不饱和脂肪酸,如DHA、EPA或花生四烯酸,和中链脂肪酸,如辛酸、癸酸或月桂酸进行调节,使得饱和脂肪酸:单不饱和脂肪酸:多不饱和脂肪酸的比例接近3:4:3。
3.碳水化合物和膳食纤维
本发明中所指的主要碳水化合物是异麦芽酮糖。其他碳水化合物实例包括糖醇(山梨糖醇、木糖醇、麦芽糖醇等)、蜂蜜、砂糖、葡萄糖、果糖和转化糖。
异麦芽酮糖是由通过α-1,6键键合的一分子葡萄糖和一分子果糖组成的二糖,是蔗糖的同分异构体,也被称为6-O-(α-D-吡喃葡萄糖基)-D-果糖、异麦芽酮糖或帕拉金糖(paratinose)。异麦芽酮糖的分子量为342.297,CAS号13718-94-0,作为甜味剂等使用。异麦芽酮糖在蜂蜜、甘蔗或类似物中含量极少。异麦芽酮糖也可以通过衍生于精蛋白杆菌(Protaminobacterrubrum)的α-葡糖基转移酶或类似物作用于蔗糖,使得α-1,2键转化为α-1,6键而产生。异麦芽酮糖的甜味接近于蔗糖,但是其甜度大概是蔗糖的一半。口服摄入的异麦芽酮糖,吸收前在消化道被异麦芽糖酶分解,和蔗糖一样消化成葡萄糖和果糖(ToshinaoGoda等.NipponEiyoShokuryoGakkaishi(inJapanese)(JournalofJapaneseSocietyofNutritionandFoodScience),Vol.36(3):169-173,1983)。当摄入异麦芽酮糖以与异麦芽酮糖的消化作用竞争时,也可被异麦芽糖酶消化的异麦芽酮糖、潘糖、异麦芽三糖或类似物被认为能抑制消化和吸收(JournalofJapaneseSocietyofNutritionandFoodScience),Vol.36(3),169-173,1983)。异麦芽酮糖的热量值为4kcal/g。在本发明中,异麦芽酮糖包含帕拉金糖浆(paratinosesyrup)、还原帕拉金糖(paratinose)、帕拉金糖淀粉糖浆和类似物。帕拉金糖淀粉糖浆是一种类似淀粉糖浆的液体,主要成分为通过异麦芽酮糖脱水缩合制备的低聚糖,如四糖、己糖或辛糖。
对于异麦芽酮糖,已知含有异麦芽酮糖等的低血糖食品组合物被用来治疗或预防恶性肿瘤疾病(国际专利申请的国家公开No.2009-530326)。
异麦芽酮糖的添加量可根据其他成分(乳蛋白水解物、发酵乳蛋白、含油酸的脂肪和油、乳磷脂、大豆卵磷脂或类似物)的添加量、摄入对象的病理、病症、年龄、体重、以及使用目的或类似情况进行适当调整。具体地,异麦芽酮糖添加量实例包括4-5g,优选5-7g,每100mL营养组合物,但不限于这些范围(JPA2009-530326)。
本发明的营养组合物可通过适当加入蛋白质、脂质和糖类来调节其热量值。本发明营养组合物的热量值实例包括,50-150千卡,且优选80-120千卡,每100mL营养组合物,但不限于这些范围。
在本发明的营养组合物中,整个营养组合物的蛋白质、脂质和糖类的能量比例,与第六次修订后的对日本人的推荐的膳食允许量和膳食参考摄取量一致。具体地,实例包括15-25%的蛋白质,20-30%的脂质和45-65%的糖类,但不限于这些范围。
本发明的营养组合物还含有膳食纤维。膳食纤维指的是食品中不能被人体消化酶水解的物质,基于亲水性分为水溶性膳食纤维和不溶性膳食纤维。对于水溶性膳食纤维,可使用不易消化的低聚糖,如乳果糖,或乳糖醇、蜜三糖。已知不易消化的低聚糖的生理功能是改善肠内环境,也就是,通过到达大肠依然未消化、有助于活化并且增殖肠道双歧杆菌属微生物,来调节肠道的功能。其他可使用的候选水溶性膳食纤维包括果胶(原果胶、果胶酯酸和果胶酸)、酶解瓜尔胶和枸子胶。
进一步,可使用的候选水溶性膳食纤维包括高分子水溶性膳食纤维,如大豆多糖增稠剂、魔芋葡甘露聚糖、海藻酸、低分子海藻酸、车前草(psyllium)、阿拉伯树胶、海藻多糖(纤维素、木质素类物质、琼脂、卡拉胶、海藻酸、岩藻多糖和海带多糖)、微生物胶(韦兰胶、凝胶多糖、黄原胶、结兰胶、葡聚糖、短梗霉多糖和鼠李聚糖胶(rhamsangum))和其他胶(源自种子的刺槐豆胶、罗望子胶和他拉胶、源自汁液的刺梧桐树胶和黄芪胶);低分子水溶性膳食纤维,如聚葡萄糖、非消化的糊精、菠萝纤维和麦芽糖醇。
不溶性膳食纤维增加大肠中不消化物质的体积,并缩短它们的通过时间。这提高了排便频率和粪便质量。不溶性膳食纤维候选实例包括纤维素、半纤维素、木质素、甲壳素、壳聚糖、大豆膳食纤维、小麦麸、松木纤维、玉米纤维、甜菜纤维和燕麦麸、小麦麸、薏仁麸、米麸、谷子、小米、稗子、普通高粱、糠谷类(Brancereals)、豆类麸皮、荞麦、芝麻麸皮、豆腐渣等。实例包括谷糠,如燕麦麸、黑麦麸、薏仁麸(teargrassbran)、米麸、普通谷子、小米,以及稗子、高粱、豆类(豆科)麸皮,假谷麸皮(pseudocerealbran),如荞麦、芝麻麸皮和豆渣。
除了以上蛋白质、脂质、糖类和膳食纤维,本发明的营养组合物可以使用水、蛋白质、糖类、脂质、维生素、矿物质、有机酸、有机碱、水果汁、调味剂、乳化剂、增稠剂、稳定剂或类似物。蛋白质实例包括动物或植物蛋白质,如全脂奶粉、脱脂奶粉、部分脱脂奶粉、酪蛋白、乳清粉、乳清蛋白、乳清蛋白浓缩物、乳清蛋白分离物、乳清蛋白水解物、α-酪蛋白、β-酪蛋白、κ-酪蛋白、β-乳球蛋白、α-乳白蛋白、乳铁蛋白、大豆蛋白质、蛋类蛋白质和肉类蛋白质,它们的分解产物;各种源自奶的成份,如黄油、乳清矿物质、奶油、乳清、非蛋白氮、唾液酸、磷脂和乳糖。营养组合物可含有肽和氨基酸,如酪蛋白磷酸肽或赖氨酸。糖类实例包括糖、加工淀粉(除糊精外,可溶性淀粉、英国淀粉(Britishstarch)、氧化淀粉、淀粉酯和淀粉醚),和膳食纤维。脂质实例包括动物油和脂肪,如猪油和鱼油,它们的分馏油,氢化油,酯交换油;植物油,如棕榈油、红花油、玉米油、菜籽油、椰子油,以及它们的分馏油,氢化油和酯交换油。维生素实例包括维生素A、胡萝卜素、复合维生素B、维生素C、复合维生素D、维生素E、复合维生素K、维生素P、维生素Q、烟酸(niacin)、烟酸(nicotinicacid)、泛酸、生物素、肌醇、咕啉和叶酸;矿物质实例包括钙、钾、镁、钠、铜、铁、锰、锌和硒。有机酸实例包括苹果酸、柠檬酸、乳酸、酒石酸和异抗坏血酸。此外,该营养组合物含有具有减小排泄物异味效果的物质(例如,食用香草提取物5mg至500mg(0.005%至0.5%)),类胡萝卜素制剂(例如,含有α-胡萝卜素、β-胡萝卜素、番茄红素或叶黄素的制剂,10μg至200μg(0.00001%至0.0002%)),或抗氧化剂(儿茶酸、多元酚或类似物)。这些成份的两种或更多可组合使用,也可使用含有大量这些成分的合成产品和/或食品。食品的形式可以是固体、液体、凝胶或类似物中的任何一种。
本发明的营养组合物可通过本领域已知的方法制得。必要时,将以上描述的部分或全部原材料混合,然后根据需要进行均匀化。均化作用指的是通过充分混合,均匀化混合好的每个成份的步骤,同时将脂肪球和其他成分的粗大颗粒机械地微粒化,用于防止脂肪形成乳皮和聚集,从而实现营养组合物的均匀乳化状态。
在本发明营养组合物的生产中,进行了加热处理或热巴氏灭菌。热巴氏灭菌条件可以是用于食品的典型的巴氏灭菌条件,热巴氏灭菌可使用普通装置实现。例如,可采用62–65℃×30分钟,72℃或更高温度×15秒或更长时间,72℃或更高温度×15分钟或更长时间,或120–150℃×1-5秒巴氏灭菌;121–124℃×5-20分钟或105–140℃下灭菌;蒸馏(retort)(高温高压)巴氏灭菌;高压蒸汽灭菌或类似方法,但不限于这些。热巴氏灭菌可以优选在加压条件下实施。
进一步,实例包括,方法之一,液体营养组合物先热灭菌,然后无菌包装入容器(例如,UHT灭菌方法和无菌包装方法的结合方法);方法之二,将液体营养组合物包装进容器,然后和容器一起热灭菌(例如,高压灭菌器法);方法之三,将组合物包装入罐容器或用于液体食物或经口/插管供给营养物的各种容器(所谓的软袋、营养袋或类似物),进行蒸馏巴氏灭菌(例如,115-145℃,5-10秒),然后在约140-145℃热灭菌约5-8秒,冷却后无菌包装,但不限于这些方法。
通过热处理或热巴氏灭菌杀死来源于原料发酵乳的发酵剂(乳酸杆菌、双歧杆菌、丙酸杆菌或类似物)。
本发明的组合物可用来抑制肿瘤生长。本发明的营养组合物以口腔/肠内营养物的形式可用于患者、老年人、婴儿或类似人群的营养管理,例如,以液体食物、经口/插管供给营养物、饮品和凝胶食品的形式。
本发明营养组合物的渗透压大约为500-1000mOsm/l,渗透压实例包括约550-750mOsm/l。在室温下测量粘度,营养组合物粘度实例包括20-100cp(1cp=1mPa·s),优选25-60cp,更优选30-50cp,但不限于这些范围。
进一步,本发明营养组合物的pH可调节至4.6或更低,优选3.0-4.3,更优选3.8-4.2,但不限于这些范围。
本发明的营养组合物能有效抑制患有肿瘤病人的肿瘤生长。更具体地,可用于抑制抑制患有结肠肿瘤、结肠实体肿瘤、肉瘤细胞肿瘤、肺癌、胸腺瘤、乳腺癌、咽癌、喉癌、胃癌、食道癌、肝癌、胆管癌、胆囊癌、胰腺癌、阴茎癌、肾盂和输尿管癌、肾细胞癌、睾丸瘤、前列腺癌、膀胱癌、外阴癌、宫颈癌、子宫内膜癌、子宫肉瘤、阴道癌、卵巢癌、卵巢生殖细胞肿瘤或脑肿瘤等病人的肿瘤生长。
在移植了肿瘤细胞株的小鼠体内实验中,发现本发明的营养组合物抑制了肿瘤增殖和对周围组织的浸润。当肿瘤浸润周围组织时,肿瘤有可能转移。当肿瘤浸润内部器官或周围神经时,引起疼痛。于是,本发明的组合物能抑制肿瘤生长、肿瘤浸润/转移或肿瘤引起的疼痛。
本发明的营养组合物在医药品或饮品或食品中的每天膳食摄入量不受限制,因为它根据摄入对象的病理、年龄、病症、体重、使用目的而变化,以及本发明的营养组合物是否是仅有的营养物等。特别地,膳食摄入量的实例包括,每人每天,400-1600mL,优选600-1600mL,更优选800-1200mL。对于一个成年人,例如,可以接受多达3000mL每天。膳食摄入量也可以由摄入对象的主治医师决定。
进一步,本发明的营养组合物可以和传统已知的具有抗肿瘤活性的医药产品或食品结合使用。
本发明的营养组合物可以以医药品或饮品或食品的任一形式使用。例如,本发明的营养组合物,当作为医药品直接服用或作为特定饮食用食品(如特定保健用食品)、营养成分功能声称食品、营养补充食品、液体食品或补品直接摄入时,预期可提高肠道菌群。或者,无论食品形态,如液体、浆体或固体、粉体,该组合物可被加入各种食品(牛奶、软饮料、发酵乳、酸奶、奶酪、面包、饼干、梳打饼、比萨饼皮、配制乳粉、液体食品、特定膳食用食品、病患食品、营养食品、冷冻食品、加工食品、其它市售食品和类似物),然后被摄取(consumed)。进一步,例如,当营养组合物以粉体形态使用时,可通过喷雾干燥或冷冻干燥的方法制得所述组合物。
本发明可以作为医药品或补品以多种形式施用,例如,当本发明的营养组合物用作医药品或补品时,它可以以不同的形式施用。其形式实例包括口服施用,如片剂、胶囊剂、颗粒剂、粉剂和糖浆剂。作为主要的药剂,每种这些制剂可依照常规方法配制,使用用于制剂的和通常可用于药物制剂技术领域的已知辅助材料,如赋形剂、粘合剂、崩解剂、润滑剂、风味剂、增溶佐剂、混悬剂或包衣剂。这些制剂可以进一步包含适量的钙。更进一步,可以加入适量的维生素、矿物质、有机酸、糖类、氨基酸、肽或类似物。
实施例
列出的本发明的实施例用来更具体地解释本发明。它们只是用来解释本发明的例证,本发明并不局限于它们。
材料和方法
小鼠的小鼠肉瘤肿瘤细胞(S-180)和小鼠的结肠肿瘤细胞(C26),这两种常用的癌细胞株用于形成S-180小鼠腹水肿瘤模型和C26小鼠实体肿瘤模型。小鼠随机分为两组,每组18-20只,雌雄各半。从接种的第一天起,使小鼠以本发明的营养组合物饮食(以下称为“试验组合物”)或对照饮食AIN-93M,自由进食14天。对于S-180荷瘤小鼠,记录生存时间作为主要指标。对于C26荷瘤小鼠,记录进食、体重、肿瘤重量和肿瘤体积;测定血清TNF-α和IL-6水平;并进行肿瘤组织病理学。
1.试验饮食
试验组合物黄色细粉末,其通过干燥表1和表3中的液体膳食组合物制得。脂质中油酸的量为总脂肪酸组成的39%,二十碳五烯酸为1.2%,二十二碳六烯酸为0.8%。
AIN-93M白色细粉末,由日本东方酵母有限责任公司制得。AIN-93M是对大鼠或小鼠营养研究有益的标准精制饮食,1993年经美国国家营养研究所(UnitedStatesNationalInstituteofNutrition)公开。AIN-93M精制饮食是用于成长期过成熟动物的标准(表4,表5)(AIN-93Mpurifieddietisstandardforaperiodofgrowthwastoomatureanimals(Table4,Table5))。
表3
用于制备试验组合物的液体膳食组合物
表4
AIN-93M
表5
试验组合物和AIN-93M的比较
2.移植的肿瘤细胞株
S-180细胞株中国上海癌症生物技术研究所提供
C26细胞株中国上海癌症生物技术研究所提供
3.动物
种类:BALB/c小鼠
来源:上海SIPPR-BK实验动物有限责任公司
性别:雌雄各半
年龄:6-7周
体重:18-20g
数量:76(36只小鼠用于S-180实验,40只小鼠用于C26实验)
动物生产许可证编号:SCXK(上海)2008-0016
动物使用许可证编号:SYXK(上海)2008-0027
居住条件:SPF实验动物房,24±1℃,相对湿度40-70%,12/12小时光/暗循环,所有小鼠自由进食。
4.方法
接种前一星期复原(recover)S-180细胞株和C26细胞株。使用BALB/c小鼠建立小鼠肿瘤模型。控制小鼠,将皮肤消毒,小鼠腹腔内接种S-180细胞或小鼠右后腋窝附近皮下接种C26细胞。在接种日,动物被随机分为AIN-93M组和试验组合物组,分开关入笼内。
36只小鼠用于S-180腹水瘤研究,每组18只。每笼有六只小鼠,笼内放置了两个特制的粉末食物给食器,每个装有大约40g的食物。小鼠可随意进食。每隔一天称量食物摄入量,每三天记录体重,和记录死亡小鼠的数量。
40只小鼠用于C26实体肿瘤研究,每组20只。每笼有五只小鼠,笼内放置了两个特制的粉末食物给食器,每个装有大约40g的食物。小鼠可随意进食。每隔一天称量肿瘤体积和食物摄入量,每三天记录体重。接种后14天,小鼠以颈脱位法处死,并收集眼部血液。解剖出肿瘤,测量肿瘤体积,称量肿瘤重量,然后,肿瘤组织固定在10%中性福尔马林中,用于病理组织学检查。
肿瘤体积计算公式:
V(cm3)=a*b2/2(a=长直径(cm);b=短直径(cm))
肿瘤生长抑制率(%)=(对照组平均肿瘤体积–试验组合物组平均肿瘤体积)×100%/对照组平均肿瘤体积,或
肿瘤生长抑制率(%)=(对照组平均肿瘤重量–试验组合物组平均肿瘤重量)×100%/对照组平均肿瘤重量
疗效标准:两组肿瘤重量(或肿瘤体积)平均值的差异是P<0.05的统计上显著性。
5.统计分析
使用统计软件SPSS11.5版进行数据分析。在S-180肿瘤研究中,两组的平均生存时间和累积生存率用存活分析对数秩检验进行比较。在C26肿瘤研究中,两组的平均肿瘤质量和平均肿瘤体积用学生t检验进行比较。P<0.05被认为是统计上显著的。
III结果
1.小鼠S-180腹水肿瘤研究
在试验组合物组的小鼠的平均存活为13.0±0.3天(平均值±标准误差),在AIN-93M组的小鼠的平均存活为12.4±0.4天。两组在平均存活方面或累计生存率方面无显著差异(P>0.05,存活分析对数秩检验)(表6,表7)。存活曲线表明,试验组合物组小鼠的存活率高于对照组,并且两条曲线在任何时间点不交叉,说明试验组合物组的小鼠的存活时间趋于长于对照组(图1,图2),尽管没有显著差异。
2.小鼠C26实体肿瘤研究
2.1食物摄入量和体重
在整个实验期间,试验组合物组的荷瘤小鼠的平均食物摄入量为2.60±0.49g/鼠/天,对照组的荷瘤小鼠的平均食物摄入量为3.37±0.45g/鼠/天(表8)。两组的食物摄入量无显著差异(P>0.05)(图2)。两组的体重变化也没有显著差异(P>0.05)(图3)。
2.2肿瘤重量和肿瘤体积
如表8所示,试验组合物组的小鼠平均肿瘤重量是1.36±0.54g,对照组的平均肿瘤重量是1.83±0.89g,有显著性差异(P<0.05)。根据肿瘤重量计算的试验组合物组的肿瘤生长抑制率是25.59%。
同样地,试验组合物组的小鼠平均肿瘤体积是1.868±0.844g,对照小鼠的平均肿瘤体积是2.582±1.055g。在第4、7、11天,测得的试验组合物组的平均肿瘤体积全部显著地大于对照组(P<0.05)。根据肿瘤体积计算的试验组合物组的肿瘤生长抑制率是27.65%(表9,图4,图5)。
2.3肿瘤病理
在肿瘤结构解剖期间,观察到试验组合物组的肿瘤完整性更好,有清晰的边缘,容易剥离。与此相反,AIN-93M组的肿瘤似乎已经侵入周围组织,很难剥离。
病理检查显示:试验组合物组的肿瘤边缘光滑,癌细胞和出血性病灶分布在边缘区域;在中心区域,深度染色细胞密集生长(图6A),这些具有深度染色细胞核的癌细胞大小和形状不同(图6B)。AIN-93M对照组的肿瘤边缘不清晰,可以看到大面积的出血(图6C);癌细胞扩散,坏死明显,且被出血包围(图6D)。
病理检查结果表明,试验组合物组的恶性肿瘤比对照组的那些侵入性更小,表明试验组合物在某种程度上对肿瘤具有抑制作用。
A.试验组合物组:试验组合物组的肿瘤边缘整齐光滑,癌细胞扩散,可以看到伊红样的均质物质(即,出血)。在中心区域可以看到深度染色癌细胞。
B.试验组合物组:在中心区域,癌细胞密集生长,细胞有不同的大小和形状,可以看到小面积均质的伊红样的物质(即,出血)。(右上方,高倍放大:各种不同形状和大小的癌细胞,细胞核被深度染色。HE×400)
C.对照组:肿瘤边缘不一致,有大面积均质的伊红样的物质(即,出血)。
D.对照组:癌细胞扩散,坏死病灶可见,且被均质的伊红样的物质(即,出血)包围。
表6
表6两组中S180荷瘤小鼠的平均存活天数
P>0.05,存活分析对数秩检验
表7
表7两组中S180荷瘤小鼠的累积存活率
P>0.05,存活分析对数秩检验
表8
表8C26肿瘤重量和肿瘤抑制率
*与对照组相比P<0.05
表9
表9不同时间下C26肿瘤体积和肿瘤抑制率
*与对照组相比P<0.05
总结
两种常用的小鼠肿瘤模型S-180腹水肿瘤模型和C26小鼠实体肿瘤模型,被用来初步评估试验组合物是否能够抑制小鼠肿瘤生长。结果表明试验组合物组的S-180荷瘤小鼠的平均生存时间没有明显长于对照组小鼠(P>0.05);在C26小鼠肿瘤模型中,试验组合物能抑制肿瘤生长。
工业应用
本发明的营养组合物对患有结肠癌实体肿瘤的病人有用,如抑制结肠癌实体肿瘤生长和降低恶性肿瘤的侵犯。
Claims (16)
1.适用于肿瘤病人的营养组合物,包含:作为蛋白质的乳蛋白水解物和发酵乳蛋白;作为脂质的含油酸的油和乳磷脂和/或大豆卵磷脂;和作为碳水化合物的异麦芽酮糖。
2.如权利要求1所述的营养组合物,其特征在于,所述乳蛋白选自由下述所组成的组:酪蛋白、乳蛋白浓缩物(MPC)、乳清蛋白浓缩物(WPC)、乳清蛋白分离物(WPI)、α-乳白蛋白、β-乳球蛋白和乳铁蛋白。
3.如权利要求1所述的营养组合物,其特征在于,在100mL所述组合物中,所述乳蛋白水解物的量为0.9g-5.0g。
4.如权利要求1所述的营养组合物,其特征在于,所述乳蛋白水解物可通过用来自地衣芽孢杆菌(Bacilluslichenifomus)的碱性蛋白酶和来自猪胰脏的胰蛋白酶,水解乳清蛋白分离物(WPI)获得。
5.如权利要求4所述的营养组合物,其特征在于,所述乳蛋白水解物是通过用分离分子量为10,000的超滤膜进一步处理获得的渗透物。
6.如权利要求1所述的营养组合物,其特征在于,所述发酵乳蛋白源自奶酪。
7.如权利要求6所述的营养组合物,其特征在于,所述奶酪是夸克奶酪。
8.如权利要求1所述的营养组合物,其特征在于,在100mL所述组合物中,所述发酵乳蛋白的量为0.5g-6.0g。
9.如权利要求1所述的营养组合物,其特征在于,在100mL所述组合物中,所述异麦芽酮糖的量为4g-15g。
10.如权利要求1所述的营养组合物,其特征在于,所述脂质中油酸的量为总脂肪酸组成的30%或更多。
11.肿瘤生长的抑制方法,所述方法包括对需要其的患者施用有效量的营养组合物,所述营养组合物包含:作为蛋白质的乳蛋白水解物和发酵乳蛋白;作为脂质的含油酸的油和乳磷脂和/或大豆卵磷脂;和作为碳水化合物的异麦芽酮糖。
12.如权利要求11所述的方法,其特征在于,所述患者患有结肠实体肿瘤。
13.如权利要求11所述的方法,其特征在于,所述组合物是口服或管饲(肠内)营养物。
14.肿瘤治疗或减小的方法,所述方法包括对需要其的患者施用有效量的营养组合物,该营养组合物包含:作为蛋白质的乳蛋白水解物和发酵乳蛋白;作为脂质的含油酸的油和乳磷脂和/或大豆卵磷脂;和作为碳水化合物的异麦芽酮糖。
15.如权利要求14所述的方法,其特征在于,所述肿瘤是结肠实体肿瘤。
16.如权利要求14所述的方法,其特征在于,所述组合物是口服或管饲(肠内)营养物。
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