CN103153326B - 含有色氨酸的肽 - Google Patents
含有色氨酸的肽 Download PDFInfo
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- CN103153326B CN103153326B CN201180048202.8A CN201180048202A CN103153326B CN 103153326 B CN103153326 B CN 103153326B CN 201180048202 A CN201180048202 A CN 201180048202A CN 103153326 B CN103153326 B CN 103153326B
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- tryptophan
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Abstract
本发明涉及用于在精神压力中或精神压力后降低饮食或食欲的组合物,所述组合物包含含有色氨酸的肽并且具有大于0.1的Trp/LNAA比例。
Description
技术领域
本发明涉及在精神压力暴露中或精神压力暴露后降低加餐。
背景技术
精神压力往往会影响食物喜好和摄入量。尽管精神压力能够引起厌食(hypophagia),但大量的研究一致表明,轻度至中度急性精神压力能增加甜的高脂肪食物的喜好和摄取量,并且精神压力引起的情绪化饮食可归因于减少的羟色胺(5-HTergic)神经传递或功能。
脑内的5-羟色胺(serotonine)水平已与情绪、机警、警觉、睡眠开始和质量、抗焦虑作用、抑郁、感情反应控制、食欲和性行为相关联。存在许多出版物,其中脑5-羟色胺水平的改变与天然氨基酸L-色氨酸(Trp或W)的可用度相关。因为这种关联,提高血浆色氨酸水平的方法受到大量关注。已有报道,每个个体1克/天左右的色氨酸量产生临床显著的作用(Markus et al.,Am.J Clin.Nutr2005;81,1026-1033)。提高血浆色氨酸水平的一种方法涉及消耗富含乳清蛋白(whey protein)α-乳白蛋白(lactalbumin)的蛋白质制剂。α-乳白蛋白制剂是可容易获得的,并具有相对高的色氨酸浓度。然而,其中提供α-乳白蛋白的途径(见例如DE4130284和JP2279700)并未考虑脑色氨酸和5-羟色胺水平的主要决定因子不只是血浆色氨酸浓度,而是所谓的Trp/LNAA比例(Fernstrom andWurtman.Science1971,173,149-152)。该Trp/LNAA比例表示血浆中色氨酸相对于大中性氨基酸(Large Neutral Amino Acid,LNAA;即酪氨酸、苯丙氨酸、亮氨酸、异亮氨酸和缬氨酸的和)的摩尔比例。这些LNAA与色氨酸竞争摄入大脑,推测是因为使用了相同的跨越血-脑屏障的转运机制。因此,提高脑色氨酸浓度的最有效途径是供应具有高Trp/LNAA比例的制备物。大量出版物例如WO02/46210涉及从α-乳白蛋白制备具有提高的Trp/LNAA比例的肽级分。
显然,游离色氨酸(即游离氨基酸)的使用会提供最简单和最便宜的用于提供具有高Trp/LNAA比例的制剂的途径。然而,在许多国家中,存在严格管制游离色氨酸供应的立法。游离色氨酸在其多种应用形式中的最大允许水平在各个国家不同。为了以更自然的方式供应额外的膳食色氨酸,近期的途径着眼于提供富含色氨酸的蛋白质。如所述的,α-乳白蛋白及其水解产物已作为提高血浆色氨酸水平的安全选择获得普及。
WO2008/052995和WO02/46210公开了一种生产组合物的方法,所述组合物包含含有色氨酸的肽且具有大于0.15的Trp/LNAA比例,其包含水解溶菌酶、优选鸡蛋溶菌酶,以制备具有5和45之间的DH的水解产物。
发明概述
本发明涉及用于口服消耗的含有色氨酸的肽或含有色氨酸的水解产物。令人惊奇的是,我们发现含有色氨酸的肽或含有色氨酸的水解产物的口服消耗导致饮食或食欲的降低,这通过数小时的减少食物的摄入来证明。因此,根据本发明的实施方式,口服摄取含有色氨酸的肽或含有色氨酸的水解产物,用来减少饮食或食欲。
根据本发明的一个方面,含有色氨酸的肽或含有色氨酸的水解产物被用于制造口服摄取的营养品或食品,来减少饮食或食欲。因此,含有色氨酸的肽或含有色氨酸的水解产物被用来减少饮食或食欲。根据本发明的另一个方面,公开了用来减少饮食或食欲的方法,其包含将含有色氨酸的肽或含有色氨酸的水解产物口服施用于需要该治疗的受试者。
特别地,本发明在精神压力(包括急性和慢性(生活)精神压力)情况下具有其效果。
本发明涉及用于在精神压力中或精神压力后降低饮食或食欲的组合物,所述组合物包含含有色氨酸的肽或含有色氨酸的水解产物并且具有大于0.1的Trp/LNAA比例。本发明还涉及在精神压力中或精神压力后得到降低的饮食或食欲的治疗,其通过口服摄取包含含有色氨酸的肽或含有色氨酸的水解产物并且具有大于0.1的Trp/LNAA比例的组合物来实现。
此外,本发明涉及用于通过口服摄取减少饮食或食欲的含有色氨酸的肽或含有色氨酸的水解产物、或包含含有色氨酸的肽或含有色氨酸的水解产物的组合物。该组合物优选地为食品或饮料、膳食补充剂、保健品或饲料或宠物食品,包含谷物棒、烘焙物品如蛋糕和曲奇、液体食物如汤或汤粉末、包括非醇饮品和醇饮品的饮料以及要被添加进饮用水和液体食物和临床食物中的液体制剂。
发明详述
人们认为精神压力以一种消极的方式影响人类的饮食行为。精神压力可改变食物摄取,这可能导致吃零食或暴饮暴食。至少两种方式的精神压力发挥重要的作用。慢性(生活)精神压力以及急性精神压力可能与更偏爱甜品和脂肪有关。其结果往往是体重增加并可最终导致肥胖的发展。
本发明提供了包含以肽形式存在的色氨酸的组合物,其非常适用于提供血浆中Trp/LNAA比例的有效提高。我们注意到包含色氨酸的二肽和三肽有利地有助于该提高。在本发明的一个实施方式中,在工业过程中用酶(预)水解溶菌酶、优选地鸡卵溶菌酶,即(鸡卵)溶菌酶优选地以水解产物的形式提供。以水解产物形式提供时,大幅促进含色氨酸的肽的胃肠吸收。本发明的水解产物的特征在于,分子的Trp/LNAA比例高于0.1、优选地高于0.15。本发明的水解产物优选地具有5和45之间、更优选地10和40之间的DH(水解度)。
因此,本发明提供了本发明的组合物(例如水溶性含色氨酸的肽)用于在摄取肽后90分钟内、优选地60分钟内、最优选地30分钟内提高血浆中Trp/LNAA比例的用途,或用于制备下述保健品组合物的用途,所述保健品组合物用于在摄取肽后90分钟内、优选地60分钟内、最优选地30分钟内提高血浆中的Trp/LNAA比例。优选地,与含色氨酸的肽同时或几乎同时消耗蛋白质或含蛋白质食物。提高的Trp/LNAA比例表示与消耗或摄取本发明的组合物之前的情况相比该比例的提高。
“蛋白质”或“多肽”在本文中被定义为包含多于30个氨基酸残基的链。
“肽”或“寡肽”在本文中被定义为通过肽键连接的至少两个氨基酸的链。术语“肽”和“寡肽”被认为是同义的(如通常所认为的那样),且每种术语可根据上下文的需要交换使用。
“水溶性”肽是在5.0的pH下可溶于水的肽。
本文中,所有(寡)肽和多肽的化学式根据通常的实践从左到右以氨基端到羧基端的方向书写。本文中使用的氨基酸的单字母代码是本领域公知的,并可见于Sambrook,et al.(Molecular Cloning:A Laboratory Manual,2nded.Cold Spring Harbor Laboratory,Cold Spring Harbor Laboratory Press,Cold Spring Harbor,NY,1989)中。
蛋白质水解产物、水解产物或经水解的蛋白质表示通过蛋白质的酶水解形成的产物;富集的水解产物是下述蛋白质水解产物级分,其例如富含选定的肽,或其中肽或多肽已从水解产物中被去除。因此,富集的水解产物优选地是肽的混合物(或肽混合物)。因此,本发明的肽混合物是至少两种,优选地至少三种,更优选地至少四种含色氨酸的肽的混合物。更优选地,混合物包含肽的群,所述群中的多于50%、优选地甚至多于60%、最优选地多于75%的存在的肽具有低于500Da的分子量。含色氨酸的肽表示包含至少一个L-色氨酸氨基酸残基的肽。WO2008/052995和WO02/46210公开了包含可用于本发明中的与肽结合的色氨酸的蛋白水解产物和组合物。
Trp/LNAA比例表示色氨酸相对于其它大中性氨基酸(LNAA;即酪氨酸、苯丙氨酸、亮氨酸、异亮氨酸和缬氨酸的总和)水平的摩尔比例。除了血浆Trp/LNAA比例外,Trp/LNAA比例仅涉及与肽结合的氨基酸。因此,游离的色氨酸、酪氨酸、苯丙氨酸、亮氨酸、异亮氨酸和缬氨酸不被考虑进Trp/LNAA比例中。
本文中,富含色氨酸的组合物指的是,包含含有色氨酸的肽或含有色氨酸的水解产物且具有大于0.1的Trp/LNAA比例的组合物。
本发中,富含色氨酸的水解产物指的是,包含含有色氨酸的肽且具有大于0.1的Trp/LNAA比例的水解产物。
与肽结合的氨基酸是属于肽的一部分的氨基酸,并且不是游离的氨基酸。
食欲被定义为进食的期望,其通过饥饿感觉来刺激。
精神压力往往会影响食物喜好和摄入量。大量研究一致表明,慢性生活精神压力、轻度至中度急性精神压力和或消极情绪能增加甜的高脂肪食物的喜好和摄取量,并且精神压力引起的情绪化饮食可归因于减少的羟色胺(5-HTergic)神经传递或功能。特别是针对这种精神压力引起的情绪化食用零食,本发明提供了减少此类吃零食行为的方法。精神压力可以来自许多不同的情况,并且可以是短暂的或长期持久的。
慢性精神压力或慢性生活精神压力是指长期的精神压力。慢性精神压力是由于尚未得到解决的情况或在解决前持续数月或甚至数年的情况的结果而发生。
急性精神压力是指短暂的精神压力。例如,当限期将至时。
有利地,在工业过程中用酶(预)水解溶菌酶(优选地鸡卵溶菌酶),即(鸡卵)溶菌酶优选地以水解产物或富集的水解产物的形式提供。以所述(富集的)水解产物形式提供时,大幅促进含色氨酸的肽的胃肠吸收。(鸡卵)溶菌酶水解产物可被分级分离,从而提高水解产物级分的色氨酸含量。该级分或富集的水解产物与分级分离前的水解产物相比优选地具有提高的Trp/LNAA比例。具有额外的游离色氨酸的水解产物或富集的水解产物的富集也形成了本发明的一部分。
根据本发明的(富集的)蛋白水解产物或由该蛋白水解产物得到的肽级分,可以以任意合适的形式使用,例如食物或饮料,如用于特殊营养用途的食物(Foods for Special Nutritional Uses)、膳食补剂、保健品或甚至用在饲料或宠物食物中。如果在食物或饮料中使用,则优选具有相对低蛋白含量的制品,从而在消耗根据本发明的制品后维持血液中高的Trp/LNAA比例。另外,优选地向含溶菌酶水解产物的食物或饮料中添加碳水化合物,以甚至进一步提高消耗后血中的高Trp/LNAA比例。合适的食物制品包括例如谷物棒、烘焙物品如蛋糕和曲奇,以及液体食物如汤或汤粉末。除了乳制品如乳和酸乳之外,其它合适的饮料包括非醇饮品和醇饮品以及要被添加进饮用水和液体食物中的液体制剂。非醇饮品优选地为矿物质水、运动饮品、果汁、柠檬水、茶、浓缩饮品如shots、能量饮品(例如含有葡糖醛酸内酯、咖啡因或牛磺酸)和碳酸饮料(例如汽水(pops)、苏打水(sodas)和可乐饮品)。用于特殊营养用途的食物包括运动食物和临床食物。术语膳食补剂在本文中使用时表示由口摄取的制品,其含有旨在补充膳食的化合物或化合物的混合物。这些制品中的化合物或化合物的混合物可包括:维生素、矿物质、草药(herbs)或其它植物和氨基酸。膳食补剂也可以是提取物或浓缩物,并可以以许多形式存在,例如片剂、胶囊、软胶囊(softgel)、胶囊(gelcap)、液体或粉末。在本文使用时,术语保健品表示同时在营养和药物两种应用领域中的有用性。根据本发明的保健品组合物可以是适合施用给动物体(包括人体)的任何形式,特别是口服施用的任何常规形式,例如固体形式如食物或饲料(添加剂/补充剂)、食物或饲料预混物、片剂、丸剂、颗粒、锭剂、胶囊和泡腾剂配制物如粉末和片剂,或是液体形式如溶液、乳剂或悬浮液,例如饮料、糊剂和油性悬浮液。掺入根据本发明的水解产物的受控(延迟)释放配制物也构成本发明的一部分。另外,可向本发明的保健品组合物中添加多维生素和矿物质补剂,以获得足量的必需营养素,所述必需营养素在一些膳食中缺失。多维生素和矿物质补剂也适用于疾病预防,和针对由于生活模式造成的营养丢失和不足提供保护。因为本发明涉及降低食欲和饮食,这可影响体重增加,本发明的组合物优选地包含少量的碳水化合物或脂肪,从而保持组合物的低热量。因此在其中使用了本发明的最终产物,通常是“轻”制品。
附图图例
图1在摄入含色氨酸水解蛋白(M)和安慰剂蛋白(K)后面试准备期间总的食物摄入量
材料和方法
材料
从Genencor(Leiden,荷兰)获得商业名称为“Protex6L”的枯草杆菌蛋白酶。
溶菌酶作为Delvozyme L(22%干物质)从DSM Food Specialities(Delft,荷兰)获得。
氨基酸分析
根据如Amino Acid Analysis System of Waters操作员手册(MilfordMA,USA)中所述的PicoTag方法进行氨基酸分析。为此,将样品干燥并使用异硫氰酸苯酯直接衍生化。如所述使用HPLC方法定量存在的经衍生化的氨基酸。因为常规酸水解期间Trp和Cys被破坏,所以使用特殊的方法定量这两种氨基酸。为了在水解期间防止Cys降解,首先使用过氧化氢将该氨基酸氧化为半胱氨酸,然后定量。色氨酸的分析以经稍微修改的Waters步骤为基础。在该步骤中,在真空下干燥肽溶液的小份试样,然后在1小时内于150℃下氮气中,在含0.2%色胺的4M甲烷磺酸中水解。使用装备有Alltech Altima C18柱和荧光检测的HPLC直接定量反应产物。
水解程度
使用快速OPA测试(Nielsen,P.M.;Petersen,D.;Dambmann,C.Improved method for determining food protein degree of hydrolysis.Journal ofFood Science2001,66,642-646),监测用多种水解混合物孵育期间获得的水解程度(DH)。
以下的实施例进一步阐述本发明。
实施例
实施例1
使用Protex水解溶菌酶来生产富含色氨酸的水解产物
使用NaOH将含10%(w/w)纯溶菌酶的溶液调节至pH8.2,并加热至52℃。通过添加25微升Protex/(g存在的蛋白质)来开始水解。在连续搅拌并维持pH在8.2情况下,继续水解5.5小时,得到无可见沉淀物的几乎澄清的溶液。在用于失活Protex活性的加热步骤后,取样用于DH分析。溶液的DH显示几乎为30%。在10kDa过滤器上超滤经热处理的溶液,得到完全澄清溶液(富含色氨酸水解产物或富含色氨酸的水解的蛋白)。
实施例2
使用富含色氨酸的水解产物减少精神压力引起的吃零食
按照双盲、安慰剂对照、交叉设计来进行本研究。在两个实验环节中,每组42名参与者来到实验室,从而在急性精神压力暴露前后以及在富含色氨酸的水解的蛋白(THP,见实施例1)摄入或安慰剂蛋白(PLC)摄入后监测他们的食物摄入量。在测试的两天,对膳食条件的顺序进行平衡抵消,采用至少一周的清除期(washout period)。
每次测试天前,指导参与者在环节前,远离酒精至少36小时并禁食12小时;只允许不含糖的水和茶。
抵达后,志愿者要么消耗THP饮品,要么消耗PLC饮品。然后,使参与者休息(看书或看电视)2小时。参与者处在单独的房间中,然后指导每个参与者准备接受一些陌生的大学工作人员组成的小组的面试。在这5分钟的准备阶段,参与者可以免费获得预先称重的部分零食,并指示他们可以想吃多少就吃多少。准备阶段后,每对参与者汇聚在一个更大的试验室内进行15分钟的精神压力诱导面试。精神压力诱导面试阶段完成后,参与者有5分钟的时间免费获取预先称重的部分零食。
饮食控制
在这两个实验环节,消耗包含富含色氨酸的卵蛋白水解产物、THP(DSM Delft,荷兰)或者包含安慰剂酪蛋白水解产物(PLC,DSMDelft,荷兰)的饮品。表1中给出了产品的主要特性。
表1:标准蛋白质(PLC)和富含色氨酸蛋白水解产物(TPH)条件的组合物。
来源 | Trp(g) | Trp(mg) | TRP/LNAA(mol/mol) |
PLC | 4g/200ml | 32 | 0.02 |
THP | 4g/200ml | 235 | 0.21 |
表2:所用蛋白质的氨基酸组成分析(profile)
食物摄入量
为了测量吃零食行为,在精神压力暴露前(准备期)后给参与者一个食品托盘;其包含预先稳重的部分零食(小糖果、饼干和坚果)。每次在准备期后以及完成公开演讲(面试)任务后,将食物容器称重,以测定食物摄入的总量。
结果
分析结果显示显著的治疗效果(P=0.037),表明与PLC(55g±50g)相比,THP(34g±27g)后食物摄入量总共降低21g(见图1),这说明在精神压力下包含含有色氨酸的肽的溶菌酶水解产物和其他含色氨酸的水解产物减少吃零食的行为。
Claims (10)
1.组合物在制造用于精神压力中或精神压力后降低饮食或食欲的药物、食物或饮料、膳食补剂、保健品或饮料或宠物食物中的用途,所述组合物包含含有色氨酸的肽并且具有大于0.1的Trp/LNAA比例。
2.如权利要求1所述的用途,其中所述含有色氨酸的肽以含色氨酸的水解产物的形式存在。
3.如权利要求1或2所述的用途,所述组合物具有至少0.15的Trp/LNAA比例。
4.如权利要求1或2所述的用途,所述组合物具有在0.15和1.8之问的Trp/LNAA比例。
5.如权利要求1或2所述的用途,其中所述精神压力为慢性精神压力或急性精神压力。
6.组合物用于在精神压力中或精神压力后降低饮食或食欲的非治疗性用途,所述组合物包含含有色氨酸的肽并且具有大于0.1的Trp/LNAA比例。
7.如权利要求6所述的组合物的用途,其中所述含有色氨酸的肽以含色氨酸的水解产物的形式存在。
8.如权利要求6或7所述的组合物的用途,所述组合物具有至少0.15的Trp/LNAA比例。
9.如权利要求6或7所述的组合物的用途,所述组合物具有在0.15和1.8之间的Trp/LNAA比例。
10.如权利要求6或7所述的组合物的用途,其中所述精神压力为慢性精神压力或急性精神压力。
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EP10186571.5 | 2010-10-05 | ||
EP10186571 | 2010-10-05 | ||
PCT/EP2011/066801 WO2012045617A1 (en) | 2010-10-05 | 2011-09-28 | Peptides containing tryptophan |
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EP (1) | EP2624850B1 (zh) |
JP (1) | JP6041242B2 (zh) |
KR (1) | KR101957531B1 (zh) |
CN (1) | CN103153326B (zh) |
BR (1) | BR112013008096A2 (zh) |
ES (1) | ES2545187T3 (zh) |
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EP2079847B1 (en) | 2006-11-02 | 2012-03-07 | DSM IP Assets B.V. | Use of lysozyme hydrolysates containing tryptophan |
ES2545187T3 (es) * | 2010-10-05 | 2015-09-09 | Dsm Ip Assets B.V. | Empleo del hidrolizado de lisozima conteniendo triptófano para disminuir el apetito durante o después de situaciones de estrés |
PL2701716T3 (pl) | 2011-04-28 | 2018-02-28 | Dsm Ip Assets B.V. | Hydrolizaty białkowe jako środki do przezwyciężania uzależnień |
JP6436312B2 (ja) * | 2012-11-02 | 2018-12-12 | ディーエスエム アイピー アセッツ ビー.ブイ.Dsm Ip Assets B.V. | トリプトファンに富んだタンパク質加水分解物の使用 |
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US20090270337A1 (en) * | 2008-04-29 | 2009-10-29 | Conopco, Inc., D/B/A Unilever | Composition Comprising Carbohydrates and Peptides which Comprise Tryptophan |
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JPH02279700A (ja) | 1989-04-19 | 1990-11-15 | Asahi Chem Ind Co Ltd | 高トリプトファン含有ペプチド |
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US6579899B1 (en) * | 1998-07-16 | 2003-06-17 | Massachusetts Institute Of Technology | Composition for treatment of stress |
DE60034074T2 (de) | 2000-12-06 | 2007-07-12 | Campina Nederland Holding B.V. | Verfahren zur herstellung tryptophanreicher peptide |
WO2004069265A1 (en) | 2003-02-07 | 2004-08-19 | Campina B.V. | Use of tryptophan rich peptides |
US7064104B2 (en) * | 2003-06-13 | 2006-06-20 | The Procter & Gamble Company | Methods of managing the symptoms of premenstrual syndrome |
EP2079847B1 (en) * | 2006-11-02 | 2012-03-07 | DSM IP Assets B.V. | Use of lysozyme hydrolysates containing tryptophan |
BRPI0911907B8 (pt) * | 2008-04-29 | 2021-05-25 | Dsm Ip Assets Bv | processo para produzir uma composição contendo triptofano e composição proteica contendo triptofano |
ES2545187T3 (es) * | 2010-10-05 | 2015-09-09 | Dsm Ip Assets B.V. | Empleo del hidrolizado de lisozima conteniendo triptófano para disminuir el apetito durante o después de situaciones de estrés |
-
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- 2011-09-28 ES ES11761592.2T patent/ES2545187T3/es active Active
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- 2011-09-28 KR KR1020137011360A patent/KR101957531B1/ko active IP Right Grant
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- 2011-09-28 CN CN201180048202.8A patent/CN103153326B/zh active Active
- 2011-09-28 WO PCT/EP2011/066801 patent/WO2012045617A1/en active Application Filing
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US20050089546A1 (en) * | 2003-09-16 | 2005-04-28 | Dick Wurtman | Compositions and methods comprising complex carbohydrate and alpha-lactalbumin |
CN101535494A (zh) * | 2006-11-02 | 2009-09-16 | 帝斯曼知识产权资产管理有限公司 | 含有色氨酸的肽 |
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BR112013008096A2 (pt) | 2016-08-09 |
WO2012045617A1 (en) | 2012-04-12 |
US20130296232A1 (en) | 2013-11-07 |
PL2624850T3 (pl) | 2015-11-30 |
ES2545187T3 (es) | 2015-09-09 |
CN103153326A (zh) | 2013-06-12 |
US9629890B2 (en) | 2017-04-25 |
KR101957531B1 (ko) | 2019-03-12 |
JP6041242B2 (ja) | 2016-12-07 |
EP2624850A1 (en) | 2013-08-14 |
JP2013541540A (ja) | 2013-11-14 |
KR20130139972A (ko) | 2013-12-23 |
EP2624850B1 (en) | 2015-07-08 |
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