CN102292114A - 通过具有从装置表面突出的排列的碳纳米管的纳米增强的装置促进的药物递送和物质传递 - Google Patents
通过具有从装置表面突出的排列的碳纳米管的纳米增强的装置促进的药物递送和物质传递 Download PDFInfo
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Abstract
本发明涉及用于在装置和组织之间的物质传递的纳米增强的装置。装置包括基材,具有被锚固在基材内的大体上排列的碳纳米管,并且碳纳米管的至少一个端部从基材突出。突出的纳米管端部可以被药物包覆,以用于药物向身体组织中的递送。本发明可以被并入血管成形术导管气囊中或并入被使用在皮肤上的贴剂中。碳纳米管可以被集合在团簇中,以有效地形成可以将流体传递至皮下组织或从皮下组织传递流体的纳米针。纳米针可以与传感器结合使用以确定诸如pH、葡萄糖水平等等的体液信息。
Description
优先权声明
本申请是要求于01/27/2009提交的标题为“CARBONNANOTUBE-BASED DEVICE FOR SUB-DERMAL BLOOD AND FLUIDPROPERTY MONITORING(用于皮下血液和流体性质监测的基于碳纳米管的装置)”的美国临时申请第61/206,071号和于05/19/2009提交的标题为“METHOD,DEVICE AND DESIGN FOR DELIVERY OF DRUGS BE ANANO-ENHANCED ANGIOPLASTY BALLOON(用于通过纳米增强的血管成形术气囊递送药物的方法、装置和设计)”的美国临时申请第61/179,639号的优先权的权益的非临时专利申请。
发明领域
本发明涉及纳米结构增强的药物递送和血液监测装置,并且更特别地涉及用于促进在装置和身体组织之间的药物或流体传递的具有从装置表面突出的排列的碳纳米管的装置。
发明背景
在皮下层中的微脉管系统,如果能接触到的话,提供有关所测量的个人的健康的生命信息。例如,血糖水平、氧含量、激素浓度等等可以直接地从血液性质测量。经常地,为了获得该信息,皮肤必须被针刺穿以将血液抽入取样装置中。这样的针的使用通常与疼痛、血液向环境的暴露或由于皮肤的破裂导致的感染的风险相关联。该程序的一种微创的替代形式是将传感器嵌入皮下层内。此外,这种获得植入物的程序(取决于规模)是有问题的,这是由于放置植入物的过程以及由患者的身体产生的潜在的排斥两者。
与本发明的主题有关的另一个领域是在血管成形术程序中。经皮腔内血管成形术(PTA)和经皮腔内冠状动脉成形术(PTCA)是已确立的、经过证明的用于以微创方式再打开狭窄的或堵塞的动脉的方法。气囊被使用导管放置在动脉的狭窄的节段中并且然后被膨胀直到内腔达到其期望的直径。使用膨胀的气囊强制地打开动脉的狭窄的部分需要非常高的压力(15bar)并且趋于导致对血管壁的损伤。身体对这样的损伤的自然响应是过度增殖,即细胞分裂的异常高的速率,其导致内腔狭窄化并且因此降低血管的功能。这与打开狭窄的或堵塞的动脉的最初目的起相反的作用。
为了对抗血管的过度增殖响应,可以使用抗增殖紫杉烷药物,例如紫杉醇。小剂量的紫杉醇与组织的单次的短时间的接触已经显示出抑制局部细胞增殖。紫杉醇是经常在癌症化学疗法中使用的有丝分裂抑制剂,因为癌细胞展示出过度增殖。紫杉醇最初地来源于太平洋紫杉树的树皮,但是现在由其他生物工程方法生产。紫杉醇的作用机理是微管的通过与微管蛋白结合的稳定化,从而干扰它们的在细胞分裂期间的正常的分解。这具有减小细胞分裂速率的净效应,并且抵制过度增殖。
抗增殖紫杉烷具有用于最小化受损的血管壁的过度增殖响应的重要性质。它们具有高亲油性(疏水性)并且紧密地结合于各种细胞成分,给予在递送部位处的良好的局部保留。虽然亲水性化合物容易地渗透入组织中,但是它们也迅速地清除。紫杉醇是疏水性化合物并且由其被递送处的内腔扩散入动脉壁中(见参考文献第1号)。
用于治疗狭窄的或堵塞的动脉的典型程序是使用紫杉醇包覆的血管成形术气囊,其中紫杉醇起在气囊打开过程之后抑制过度增殖的作用。对这一过程的主要限制如下:
1.需要长充气时间,以确保紫杉醇向动脉壁的扩散是足够的,这可能导致过度的动脉壁损伤。以前的研究显示,为了将紫杉醇的初始剂量的约90%传递至动脉壁,需要40-60分钟(见参考文献第1号);
2.为了获得过度增殖抑制的最优的益处,重要的是,紫杉醇包覆层在前进进入动脉的狭窄的节段时不会被血流损失或洗掉。以前的研究显示,紫杉醇被血流从未充气的气囊洗掉的速率是高至初始剂量的1.2%每分钟(见参考文献第2号);
3.紫杉醇包覆层的潜在的损失,这是由于在前进进入动脉的狭窄的节段时与健康的动脉壁接触;以及
4.可以被施用于单一气囊上的紫杉醇的最大剂量是约11mg(10μg/mm2),这显著地小于由FDA批准的对于紫杉酚的紫杉醇剂量(见参考文献第1号)。
应当注意,碳纳米管用于增强气囊导管的材料性质的目的的用途已经在美国专利第7,037,562号中提出。然而,在该专利中,将碳纳米管与用于加强材料的目的而并非用于药物递送目的的基体材料混合。此外,碳纳米管未被以使得它们将从表面突出并且促进药物递送的方式排列或部分地锚固在基体材料中,使装置对该目的不起作用。
因此,持续存在对保护所抽出的流体免受大气压力的微创血液监测装置的需求,且还持续存在对可以将大量的期望的药物有效地在相对短的时期内递送至被影响的部位且最少的药物损失到经过的体液的药物递送系统的需求。
所引用的参考文献的列表
在整个本申请中引用以下的参考文献。为了清楚和便利,参考文献在本文中列出,作为阅者的中心资源。以下的参考文献在此以引用方式并入,如同被完全地包括在本文中一样。参考文献通过参考相应的参考文献号而被在本申请中引用。
(1)Creel,C.J.,M.A.Lovich和E.R.Edelman,Arterial paclitaxeldistribution and deposition(动脉紫杉醇分布和沉积).Circulation Research,2000,86(8):第879-884页。
(2)Scheller,B.,U.Speck,C.Abramjuk,U.Bernhardt,M.Bohm和G.Nickenig,Paclitaxel balloon coating,a novel method for prevention andtherapy of restenosis(紫杉醇气囊包覆层,一种新颖的用于再狭窄的预防和治疗的方法).Circulation,2004.110(7):第810-814页。
(3)Bronikowski,M.J.,Longer nanotubes at lower temperatures:Theinfluence of effective activation energies on carbon nanotube growth bythermal chemical vapor deposition(在较低温度的较长纳米管:有效活化能对由热化学气相沉积生长的碳纳米管的影响).Journal of PhysicalChemistry C,2007.111(48):第17705-17712页。
发明概述
本发明涉及纳米增强的装置,其用于装置和组织之间的物质传递。在一个方面,装置包括基材以及大体上排列的具有两个端部的碳纳米管的阵列,碳纳米管被锚固在基材内且至少一个端部从基材突出,由此突出的碳纳米管增强装置的物质传递能力。
在装置的另一个方面,碳纳米管阵列被选自由药物和基因组成的组的物质包覆。
在装置的又一个方面,被锚固的碳纳米管的突出的端部不具有包覆药物并且药物包覆区域由选自由碳纳米管的侧壁和在碳纳米管之间的自由空间组成的组的区域组成。
在另一个方面,装置类型选自由血管成形术气囊和待用在组织上的贴剂组成的组。
在装置的另外的方面,包覆药物是紫杉醇。
在装置的另一个方面,药物从装置向组织的递送通过选自由以下组成的组的方法来增强:对碳纳米管阵列的至少一部分的直接的或间接的加热、使电流通过碳纳米管阵列的至少一部分、对碳纳米管阵列的至少一部分的激光刺激或其他光学刺激、在碳纳米管阵列的至少一部分上的超声波、碳纳米管阵列的至少一部分的机械振动以及在制造期间改变纳米管阵列的疏水性。
在又一个方面,药物递送增强在选自由以下组成的组的时间被执行:立即地和根据定时程序。
在本发明的装置的另一个方面,碳纳米管被布置成一个或多个团簇,使得每个团簇有效地起针的作用。
在装置的另一个方面,基材包括多孔材料,由此物质可以通过碳纳米管的固有的毛细管作用而被传递至装置和组织或被从装置和组织传递。
在另一个方面,多孔基材材料负载有待通过碳纳米管传递至组织的物质。
在装置的又一个方面,碳纳米管团簇以使得碳纳米管团簇被彼此电隔离的方式被图案化,并且其中装置还包括用于读取纳米管团簇的电势的传感器。
在另一个方面,碳纳米管的固有的电导率通过用预涂覆的化学品处理碳纳米管来改变。
在本发明的另外的方面,碳纳米管团簇的子集(subset)用葡萄糖氧化酶处理,并且碳纳米管的另一个子集用铁氰化物处理,由此通过血液的传导可被实现并且允许血糖水平的测量。
附图简述
结合参考以下附图,从以下的对本发明的各种方面的详细描述,本发明的目的、特征和优点将是明显的,在附图中:
图1是具有从其突出的碳纳米管的聚合物锚固层的侧视图;
图2A是根据本发明的纳米增强的装置的侧视图,描绘了碳纳米管的仅一个端部从基材突出;
图2B是根据本发明的纳米增强的装置的侧视图,描绘了装置刺破组织的表面;
图3A是根据本发明的纳米增强的装置的侧视图,描绘了碳纳米管被药物或剂包覆;
图3B是根据本发明的纳米增强的装置的侧视图,描绘了碳纳米管阵列的内部部分被药物包覆,但是纳米管端部被保留不具有包覆药物;
图4A是根据本发明的被锚固在多孔基材材料中的纳米针的侧视图;
图4B是具有从其突出的多重纳米针的装置的侧视图;
图5A是连接于传感器以形成纳米探针的纳米增强的装置的侧视图;
图5B是根据本发明的纳米探针装置的俯视图,其中纳米针团簇被图案化为电隔离的子集;
图6A是示出了根据本发明的纳米增强的导管气囊的图示,其中气囊处于放气状态中;
图6B是示出了根据本发明的纳米增强的导管气囊的图示,其中气囊处于膨胀状态中;
图7A是根据本发明的纳米增强的表皮贴剂的透视图;以及
图7B是示出了被附着在使用者的臂上的纳米增强的表皮贴剂的图示。
详细描述
本发明涉及纳米结构增强的药物递送和血液监测装置,并且更特别地涉及用于促进在装置和身体组织之间的药物或流体传递的具有从装置表面突出的排列的碳纳米管的装置。以下的描述被呈现以使本领域的普通技术人员能够制造和使用本发明并且能够将其结合入特定的应用的情况中。各种修改,以及在不同的应用中的多种用途,对于本领域的技术人员来说将是易于明白的,并且本文定义的一般原理可以被应用于各种实施方案。因此,本发明不意图被限制于所呈现的实施方案,而是将符合与本文公开的原理和新颖的特征一致的最宽的范围。
在以下的详细描述中,阐明了多种具体的细节,以提供对本发明的更彻底的理解。然而,对本领域的技术人员将明显的是,本发明可以被实施,而不一定被限于这些具体的细节。在其他情况下,熟知的结构和装置被以框图的形式而非详细地示出,以避免模糊本发明。
阅者的注意力被引导至与本说明书同时提交的并且与本说明书共同向公众检查开放的所有论文和文献,并且所有这样的论文和文献的内容都被以引用方式并入本文。在本说明书(包括任何伴随的权利要求、摘要和附图)中公开的所有特征都可以被起到相同的、等效的或相似的目的的替代特征代替,除非另外明确地说明。因此,除非另外明确地说明,否则所公开的每个特征都仅是一个通用系列的等效的或相似的特征的一个示例。
此外,权利要求中的不明确地声明用于进行具体的功能的“手段”或用于进行具体的功能的“步骤”的任何要素都不应被解释为35U.S.C.第6节第112款中规定的“手段”或“步骤”条款。特别地,在本文权利要求中的“的步骤”或“的行为”的使用不意图调用35U.S.C.第6节第112款的条款。
(1)引言
本发明涉及纳米结构增强的药物递送和血液监测装置,并且更特别地涉及用于促进在装置和身体组织之间的药物或物质传递的具有从装置表面突出的排列的碳纳米管的装置。
碳纳米管技术的最新进展使创造具有在纳米(nm)数量级的直径和在毫米数量级的长度的针或棒成为可能。碳纳米管在本申请中是有益的,因为神经在真皮层内的分布与具有相似于碳纳米管的尺寸的纳米尺度针相比是稀疏的。例如,紧密接触的200个具有10nm直径的碳纳米管的聚集体将使整个聚集体占据小于2微米的总直径。美国专利申请公布第2008/0145616A1号描述了用于选择性地锚固大量纳米尺度结构的方法。这种锚固方法允许控制将纳米尺度结构锚固入其他材料中的深度。
(2)本发明的细节
碳纳米管可以以多种方式来形成,最简单的方式是使用催化剂包覆的基材的热化学气相沉积。通常,几纳米的铁被包覆至硅上,被预先制备并且在含碳原料气流例如乙烯下被放置在管式炉中,并且升温至合适的温度,例如725摄氏度。碳纳米管的热化学气相沉积生长(见参考文献第3号)生成在生长基材上的竖直地排列的碳纳米管,生长基材通常是硅。
将碳纳米管的阵列锚固在基材上可以通过使碳纳米管的阵列与未固化的聚合物材料层接触并且然后经历固化步骤来进行,如在上文引用的美国专利申请公布第2008/0145616 A1号中描述的。为了将碳纳米管锚固入管状基材例如气囊材料中,可以使用围绕中心棒以保持圆柱形形状的牺牲释放层。首先,牺牲释放层被沉积并且围绕棒形成并且尽可能地固化,并且然后气囊材料围绕释放层沉积。然后纳米管被附接入或锚固入气囊材料中。最后,牺牲释放层被除去,释放具有被附接入或锚固入其中的纳米管的气囊。
在碳纳米管被附接于基材之前或之后,药物、基因或其他物质可以在另一个步骤中被包覆至碳纳米管上。这在使用药物紫杉醇时是特别可行的,因为紫杉醇是疏水性的,并且碳纳米管在它们的生长状态中也是疏水性的。因此,紫杉醇分子将优先附着于碳纳米管,直到碳纳米管被放置为与血管中的堵塞材料紧密接触,堵塞材料通常是脂肪性的并且在本质上是疏水性的。此外,因为碳纳米管表面在本质上是疏水性的,所以残留在纳米管的层内的任何物质将免除接触纳米管表面的水和水溶液以及混合物,例如血液。因此,在本发明的期望的方面,碳纳米管的末端被保留不具有药物包覆层,并且药物包覆层被限于纳米管的侧壁和/或在纳米管之间的自由空间。该方面提供安全的表面接触并且还避免将药物过早释放入非目标组织或体液中。
通过碳纳米管向目标组织中的药物释放可以通过与能够进行多种增强方法的增强装置结合来增强,增强方法包括但不限于将以下中的一个或多个施用于碳纳米管:直接的或间接的加热,例如通过诸如射频(RF)波的电磁辐射;电流;激光或其他光学刺激;超声波;机械振动;以及通过在制造期间改变纳米管阵列的疏水性(这可以例如通过使用氧等离子体处理来进行)。这些增强方法可以被施用于碳纳米管的空间图案化的部分或完全的集合。方法还可以立即地或根据定时程序被施用。
图1是示出了本发明的一般的实施方案的图示。装置包括基材层100以及多个大体上排列的碳纳米管102。每个碳纳米管具有两个端部104,并且碳纳米管102被锚固在基材100内,且至少一个端部104从基材突出。在所示的实例中,碳纳米管102的两个端部从基材100突出。
图2A图示了装置的另一个实施方案,其中排列的碳纳米管102的仅一个端部104从基材100突出。图2B示出了碳纳米管阵列102的端部104刺破身体组织200例如皮肤、动脉壁、结肠等等的表面。应当注意,本发明还具有在医学领域之外的潜在的用途。装置可以可行地向除了身体组织之外的各种目标传递物质,并且因此不被限制于上文的应用。
碳纳米管可以被药物、基因或待被传递至组织的其他物质包覆。图3A图示了碳纳米管102被待被传递至身体组织的物质300包覆的装置的实施方案。在如图3B中所示的可选择的实施方案中,碳纳米管的端部104被保留不具有包覆物质300。包覆物质300,在这种情况下,将被限于碳纳米管的侧壁和/或在碳纳米管之间的空间。
(3.0)具体应用
(3.1)针
本发明的一个方面是使用碳纳米管的图案化分布以起微尺度针的作用的装置,微尺度针可以容易地刺穿真皮层,到达微脉管系统或孔隙流体。这样的装置可以被用作针或反向针(reverse needle)以分别促进物质向身体的传递和从身体的传递。图4A示出了碳纳米管102的群被锚固入基材100中使得它们形成纳米针。在所示的实施方案中,基材是多孔的400。双面箭头402示出了物质通过碳纳米管102向基材或从基材迁移的可能的方向。当起针的作用时,血液或孔隙流体通过基于碳纳米管的针与流体的皮下接触被抽入多孔基材400中,所述皮下接触相似于图2B中示出的刺破行为。流体的分析通过一些单独的方法来进行,例如通过光学分析。为了起反向针的作用,多孔基材400使用一些剂例如药物预负载,然后所述剂可以通过基于碳纳米管的针被递送入皮下区域中。图4B示出了具有纳米管102的多重针状团簇的装置。
(3.2)探针
纳米针装置可以与传感器组合以起到用于监测血糖水平或其他血液或体液性质例如pH、糖水平、氧含量等等的探针的作用。装置基于碳纳米管的固有的电导率起作用。图5A是根据本发明的纳米探针的图示。探针包括被锚固在与传感器500结合的基材100中的一个或多个碳纳米管102团簇。传感器可以操作地连接于碳纳米管102或连接于基材100,或二者,这取决于期望的操作模式。在图5A中,传感器500操性地连接于碳纳米管102团簇。在如图5B中的俯视图中所示的期望的实施方案中,纳米管团簇以使得它们被彼此电隔离的方式被图案化。所示的图案包括纳米管团簇的内环502和外环504。图5B中所示的方面的一个具体的用途是测量血糖水平。在该应用中,碳纳米管团簇的子集(例如内环502)被葡萄糖氧化酶处理,而碳纳米管团簇的另一个子集(例如外环504)被铁氰化物处理,由此通过血液的传导性可被实现并且允许血糖水平的测量。应当注意,本发明可以采用除了图5B中所示的具体实施方式之外的许多纳米管团簇的图案。此外,装置还可以用作包覆的探针,其中诸如药物的剂可以被预包覆至碳纳米管的表面上或碳纳米管内,如图3A和图3B中所示的,并且因此通过碳纳米管的穿透而被递送和释放入身体组织中。
(3.3)漆刷物
如图4B中所示的碳纳米管的组件还可以借助于碳纳米管的固有的毛细管作用充当湿漆刷物。相似的方法在美国专利申请第11/124,523号中描述,其据此以引用方式并入,如同在本文中完整地阐明一样。
(3.4)血管成形术气囊
本发明的另一个期望的实施方案是在血管成形术程序中用于递送药物紫杉醇以在气囊的充气以打开血管的一部分之后预防组织细胞的过度增殖。然而,本发明不应当被解释为限于在血管成形术程序中的使用或限于紫杉醇的递送。本发明可以可行地用于向身体的各部分递送种类繁多的物质,身体的各部分包括但不限于皮肤、子宫、支气管以及包括结肠的胃肠道的各部分,。
图6A和图6B图示了根据本发明的纳米增强的血管成形术气囊600。图6A示出了以放气形式的血管成形术气囊,如其将被插入血管或其他身体包封物中。当充气时,如图6B所示,表面碳纳米管102以图2B所示的方式刺穿血管表面,将包覆在其上的任何药物释放入目标组织中。
本发明,关于在血管成形术气囊中的用途方面,克服了如在上文的背景部分中描述的相关技术的限制,这些限制包括长充气时间、由于在插入过程期间的损失导致的可用的药物量的减少、向健康组织过早的递送和/或非期望的递送、以及低的总递送药物量。因为用于递送的药物的可能的量取决于表面积,所以纳米增强的气囊-导管将在插入过程中保持更大的药物量并且以1000倍的数量级的因子增加最大可递送药物量,与由纳米结构提供的表面积的增加相似。
以下的表达式比较了纳米增强的气囊与不具有纳米增强的标准气囊的表面积:
其中:
d(图1中的106)是单一碳纳米管的直径;
h(图1中的108)是单一碳纳米管的长度;
s(图1中的110)是在单一碳纳米管之间的距离;
R(图6B中的604)是气囊的半径;并且
L(图6A中的602)是气囊的长度,并且
使用以下假定的典型值:
d=10nm;
s=100nm;并且
h=500nm,
该结果表明如所描述的导管气囊的表面扩散增强可以容易地将表面积增加103的因子。
(3.5)表皮贴剂
本发明的纳米增强的表面可以被结合入待被用在皮肤上的表皮贴剂。图7A图示了纳米增强的表皮贴剂700的实例。贴剂具有从待被附着于皮肤的表面突出的排列的碳纳米管102。贴剂700的一部分,例如周边702,可以被粘合剂包覆,以利于向皮肤的粘附。当贴剂700被施用于皮肤时,如图7B中,碳纳米管102将以图2B中所示的方式进入皮肤并且将包覆在其上的任何药物或剂释放入皮肤中。贴剂可以与本发明的在前描述的实施方案中的任一个结合使用。
Claims (13)
1.一种纳米增强的装置,用于所述装置和组织之间的物质传递,包括:
基材;以及
大体上排列的具有两个端部的碳纳米管的阵列,所述碳纳米管被锚固在所述基材内且至少一个端部从所述基材突出,由此突出的碳纳米管增强所述装置的物质传递能力。
2.根据权利要求1所述的装置,其中所述碳纳米管阵列被选自由药物和基因组成的组的物质包覆。
3.根据权利要求2所述的装置,其中所述碳纳米管具有侧壁并且其中所述纳米管阵列含有在所述碳纳米管之间的自由空间,并且其中所锚固的碳纳米管的突出的端部不具有包覆物质,并且物质包覆区域由选自由所述碳纳米管的侧壁和在所述碳纳米管之间的自由空间组成的组的区域组成。
4.根据权利要求2所述的装置,其中所述装置选自由血管成形术气囊和待用在所述组织上的贴剂组成的组。
5.根据权利要求4所述的装置,其中所述药物是紫杉醇。
6.根据权利要求2所述的装置,其中所述药物从所述装置向所述组织的递送通过与能够进行选自由以下组成的组的增强行为的药物递送增强装置结合来增强:对所述碳纳米管阵列的至少一部分的直接的或间接的加热、使电流通过所述碳纳米管阵列的至少一部分、对所述碳纳米管阵列的至少一部分的激光刺激或其他光学刺激、在所述碳纳米管阵列的至少一部分上的超声波、所述碳纳米管阵列的至少一部分的机械振动以及在制造期间改变所述纳米管阵列的疏水性。
7.根据权利要求6所述的装置,其中所述药物递送增强装置在选自由以下组成的组的时间执行所述增强行为:立即地和根据定时程序。
8.根据权利要求1所述的装置,其中所述碳纳米管被布置成一个或多个团簇,使得每个团簇有效地起针的作用。
9.根据权利要求8所述的装置,其中所述基材包括多孔材料,由此物质能通过所述碳纳米管的固有的毛细管作用而被传递至所述装置和所述组织或被从所述装置和所述组织传递。
10.根据权利要求9所述的装置,其中多孔基材材料负载有待通过所述碳纳米管传递至所述组织的物质。
11.根据权利要求8所述的装置,其中所述碳纳米管的团簇以使得所述碳纳米管的团簇被彼此电隔离的方式被图案化,并且其中所述装置还包括用于读取所述纳米管的团簇的电势的传感器。
12.根据权利要求11所述的装置,其中所述碳纳米管具有固有的电导率,并且其中所述碳纳米管的所述固有的电导率通过用预涂覆的化学品处理所述碳纳米管来改变。
13.根据权利要求12所述的装置,其中所述碳纳米管的团簇的一子集用葡萄糖氧化酶处理,并且所述碳纳米管的另一个子集用铁氰化物处理,由此通过血液的传导可被实现并且允许血糖水平的测量。
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EP2381972A2 (en) | 2011-11-02 |
US20100196446A1 (en) | 2010-08-05 |
JP2012516202A (ja) | 2012-07-19 |
US20150238742A1 (en) | 2015-08-27 |
WO2010087971A3 (en) | 2010-12-23 |
WO2010087971A2 (en) | 2010-08-05 |
JP5620408B2 (ja) | 2014-11-05 |
US9050444B2 (en) | 2015-06-09 |
US20130158377A1 (en) | 2013-06-20 |
US9352136B2 (en) | 2016-05-31 |
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