CN112689517A - 提高抗癌药剂效能的方法 - Google Patents

提高抗癌药剂效能的方法 Download PDF

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CN112689517A
CN112689517A CN201980050535.0A CN201980050535A CN112689517A CN 112689517 A CN112689517 A CN 112689517A CN 201980050535 A CN201980050535 A CN 201980050535A CN 112689517 A CN112689517 A CN 112689517A
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让·德冈兹伯格
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Abstract

本发明涉及一种提高抗癌药剂的治疗效能的方法，所述方法包括向需要的对象给药有效量的吸附剂或抗生素失活酶。

Description

提高抗癌药剂效能的方法
技术领域
本发明涉及疗法领域。更具体来说，在本文中公开了一种提高抗癌药剂的治疗效能的方法，所述方法包括向需要的对象给药有效量的吸附剂。具体来说，本发明可用于在对象中防止肠道微生物菌群的破坏，并因此提高给药到所述需要的对象的抗癌药剂的治疗效能。
背景技术
癌症的特征在于身体中细胞的不受控制的生长，导致侵入必需器官并常常导致死亡。最初，癌症的药理学治疗利用靶向包括正常细胞在内的所有快速分裂的细胞的非特异性细胞毒性药剂。这些非特异性细胞毒性药剂具有抗肿瘤效果，但它们的使用通常受到严重毒性的限制，并且它们通常不能以持久的方式治愈患者。随着对能够使癌细胞茁壮成长的蛋白质和途径的理解的演变，已经开发出阻断癌细胞中被激活的特定蛋白质的更新的、靶向性更强的药剂。
具体来说，开发了免疫肿瘤药剂：它们利用患者的免疫系统来帮助治疗癌症。免疫系统具有特异性破坏肿瘤而对正常组织没有毒性以及可以以长期持续的方式阻止癌症复发的长期免疫的最大潜力。然而，此类药剂的效能可以被提高。
最近已显示，微生物菌群的组成对抗癌免疫监督的有效性具有重大影响，因此可能对靶向细胞毒性T淋巴细胞蛋白4(CTLA-4)或程序化细胞死亡蛋白1(PD-1)/程序化细胞死亡1配体1(PD-L1)轴的免疫检查点抑制剂的治疗活性以及免疫原性化学疗法的活性有贡献(Routy等，Nat Rev Clin Oncol.2018Jun；15(6):382-396)。
在更普遍的情况下，已将微生物菌群的改变与化疗效能受损相关联。具体来说，抗革兰氏阳性细菌的抗生素可能对某些化疗药剂例如环磷酰胺或顺铂的抗癌活性具有负面影响(Pflug,等，Oncoimmunology,2016 Apr 22；5(6):e1150399)。
所述微生物菌群的破坏通常被称为生态失调(dysbiosis)，并且可以通过所述微生物菌群的多样性降低和组成的改变来表征。除了其他分子之外，最近已显示抗生素严重破坏微生物菌群，并且所述破坏可以持续到抗生素摄入后几个月。
因此，在接受或即将接受抗癌药剂、特别是免疫肿瘤药剂的患者中，提供用于防止由使用引起生态失调的药剂例如抗生素引起的肠道微生物菌群的破坏的解决方案以提高它们的效能，并且除了其他临床相关结果之外还提高患者的无肿瘤进展生存率和总体生存率，将是有利的。
发明内容
本发明涉及一种在需要的对象中提高抗癌药剂的效能的方法，所述方法包括向所述对象给药有效量的吸附剂。本发明还涉及用于提高抗癌药剂的效能的吸附剂。本发明还涉及一种吸附剂，其与抗癌药剂例如免疫肿瘤药剂相组合用于治疗或预防癌症的方法中。
由于本发明，抗癌药剂的效能得以提高。具体来说，免疫肿瘤药剂的效能得以提高。不希望受到任何理论限制，据信这种提高是由于保留了肠道的共生微生物菌群，从而保留了抗癌免疫监督并甚至加强了针对癌症的免疫活性。
在特定实施方式中，所述对象是哺乳动物对象，优选为人类对象.
在其他特定实施方式中，所述对象已接受、接受或将接受引起生态失调的药剂。在特定实施方式中，所述引起生态失调的药剂是给药到所述对象用于预防或治疗感染的抗生素。在这种情形中，给药所述吸附剂是为了防止所述抗生素对肠道中、特别是肠的下部例如末端回肠、盲肠或结肠中的共生微生物菌群可能具有的不良作用。
在另一个实施方式中，所述对象不接受抗生素治疗。在这种情形中，给药所述吸附剂是为了防止由于抗生素给药之外的其他原因引起的肠道共生微生物菌群的破坏。例如，所述吸附剂可用于通过直接影响有害细菌例如艰难梭菌(Clostridium difficile)的生长或通过吸附由这些有害细菌释放的毒素，来治疗由所述有害细菌引起的感染。在另一个实例中，所述吸附剂可用于减轻某些治疗的副作用，所述治疗使用给药到所述患者的可能对肠道微生物菌群具有有害效应的药理学药剂或其他药剂。
在特定实施方式中，所述吸附剂是活性炭。
另一方面，所述对象可以用抗生素失活酶代替吸附剂来给药。在其他特定实施方式中，所述抗生素失活酶是β-内酰胺酶。在其他特定实施方式中，所述抗生素失活酶是红霉素酯酶。
在其他特定实施方式中，所述吸附剂或抗生素失活酶用于口服给药。
在另一个特定实施方式中，所述吸附剂或抗生素失活酶是在制剂中，所述制剂在肠的所需部分中，特别是在肠的下部、特别是末端回肠、盲肠或结肠中释放所述吸附剂或抗生素失活酶。
在某些实施方式中，所述抗癌药剂可以选自但不限于：
-微管蛋白毒素，紫杉烷，例如多西他赛、紫杉醇，
-铂化合物，例如顺铂、卡铂、奥沙利铂，
-干扰DNA复制的药剂，例如DNA嵌入剂例如蒽环霉素，
-拓扑异构酶抑制剂，例如依托泊苷，
-抗代谢物，例如甲氨蝶呤、阿糖胞苷(ara-C)、吉西他滨、5-氟尿嘧啶，
-烷化剂，例如氮芥、美法仑、卡莫司汀、异环磷酰胺或环磷酰胺，
-靶向药剂，例如酶抑制剂，特别是激酶抑制剂例如埃罗替尼、索拉非尼、伊马替尼或蛋白酶体抑制剂例如硼替佐米、卡非佐米、伊沙佐米，
-靶向生长因子受体的细胞外区域的单克隆抗体，例如曲妥珠单抗、贝伐单抗和西妥昔单抗，
-免疫肿瘤药剂，例如PD-1或PD-L1抑制剂例如派姆单抗、纳武单抗、德瓦鲁单抗、阿特珠单抗、阿维单抗、德瓦鲁单抗或靶向CTLA-4的药物例如伊匹单抗，以及
-它们的组合，特别是化学治疗剂与免疫肿瘤药剂的组合。
在某些实施方式中，所述免疫肿瘤药剂可以选自但不限于：
-免疫检查点抑制剂例如PD-1抑制剂，例如纳武单抗或派姆单抗，
-PDL-1抑制剂，例如阿特珠单抗、阿维单抗或德瓦鲁单抗；或CTLA-4抑制剂，例如伊匹单抗，
-癌症疫苗，例如西普鲁塞-T，
-免疫调节剂，例如沙利度胺、来那度胺、泊马度胺，
-非特异性免疫疗法药剂，例如干扰素或白介素，
-嵌合抗原受体(CAR)-T细胞疗法，例如tisagenlecleucel或axicabtageneciloleucel，以及
-它们的组合。
发明详述
吸附剂和吸附剂制剂
术语“吸附剂”是指通常可以通过其表面与待吸附物质之间的物理化学结合来吸附所述物质的任何化合物或材料。吸附剂可以是特异性或非特异性的。用于本发明的优选吸附剂是药用级吸附剂，最好适合于在人类或动物中用于制药或兽医应用。
适合用于本发明的吸附剂的实例包括但不限于活性炭(也被称为活性碳)；粘土，包括膨润土、高岭土、蒙脱土、硅镁土、多水高岭土、锂皂土等；二氧化硅，包括胶体二氧化硅(例如
Figure BDA0002921556420000051
AS-40)、中孔二氧化硅(MCM41)、煅制二氧化硅、沸石等；滑石；消胆胺等；聚苯乙烯磺酸盐等；单磺化和多磺化树脂；以及其他树脂，例如用于细菌学检验的树脂例如
Figure BDA0002921556420000052
树脂。
优选的吸附剂是药用级活性炭(例如来自于Chemviron、Cabot、Norit、JacobiCarbons、Merck Millipore、Sigma Aldrich、Desotec、Haycarb、Donau Carbon或其他来源)。在特定实施方式中，所述吸附剂是活性炭，更特别是具有高于600m2/g、特别是高于800m2/g、特别是高于1000m2/g、特别是高于1200m2/g、特别是高于1400m2/g、特别是高于1600m2/g、甚至更特别是高于1800m2/g的比表面积的活性炭。所述活性炭可以是植物、矿物或合成来源的，它的表面任选地通过物理或化学处理而改性。在特定实施方式中，所述活性炭是植物来源的。在特定实施方式中，所述活性炭源自于泥炭。在特定实施方式中，所述活性炭源自于椰子壳。在特定实施方式中，所述活性炭源自于混合在一起的不同来源例如泥炭和椰子壳。在特定实施方式中，所述活性炭的特征在于优选地高于100、甚至更特别地高于200、甚至更特别地高于300、甚至更特别地高于400、甚至更特别地高于500、甚至更特别地高于600的欧洲糖蜜值(应该注意的是，所述欧洲糖蜜值与北美糖蜜值反相关)。在特定实施方式中，所述活性炭具有高于10g/100g、甚至更特别地高于20g/100g、甚至更特别地高于30g/100g、甚至更特别地高于40g/100g、甚至更特别地高于50g/100g、甚至更特别地高于60g/100g的安替比林值(按照欧洲药典(EU Pharmacopeia)测量)。在特定实施方式中，所述活性炭的特征在于具有0.05至0.8之间、甚至更特别地0.1至0.6之间、甚至更特别地0.15至0.5之间、甚至更特别地0.2至0.4之间的密度。
在本发明的方法中使用的吸附剂的量可以随着待处理的宿主/材料和所述吸附剂的总容量、吸附力和选择性而变。通常，吸附剂的量是足以提高抗癌药剂的效能的量。在特定实施方式中，吸附剂的量是足以防止物质例如抗生素对肠道微生物菌群的被称为“生态失调”或肠道微生物菌群破坏的有害影响的量。具体来说，吸附剂的量是足以在对象中提高免疫肿瘤药剂的效能或提高抗癌免疫监督的有效性的量。
所述用于本发明的吸附剂可以配制成组合物例如药物组合物，所述组合物可以包含可药用赋形剂、载体和/或添加剂。此类组合物包括用于口服递送、直肠递送、局部施用、粘膜施用、吸入等的制剂。在特定实施方式中，所述吸附剂被配制成适合于给药到人类或动物的药物组合物。更优选地，所述吸附剂被配制成适合于在肠中或与肠道细菌接触时，特别是在胃肠道中、更特别是在肠的下部即末端回肠、盲肠和/或结肠中释放出所述吸附剂的口服制剂。
短语“可药用”在本文中用于指称那些在健全的医学判断范围内适合与人类和动物的组织相接触使用而没有过度毒性、刺激性、过敏反应或其他问题或并发症，与合理的利益/风险比相称的化合物、材料、组合物和/或剂型。适合于吸附剂的肠道递送的制剂的实例已描述在WO2006/122835和WO2007/132022中。在另一个实施方式中，所述吸附剂被配制在核心中。优选地，吸附剂的量在所述核心的总重量的约60％至约100％之间，更优选地约70％至约98％之间，更优选地约75％至约95％之间，更优选地约80％至约90％之间。在优选实施方式中，所述吸附剂用卡拉胶配制，优选地配制成球丸的形式，正如在WO2011/104275中提出的。这种制剂可以形成核心。这种核心可以用包衣层覆盖，使得所述吸附剂在肠的下部中，即在末端回肠、盲肠和/或结肠中释放。或者，可以将多个核心包含或包埋在适合于在肠的下部中，即在末端回肠、盲肠和/或结肠中释放其内含物的剂量单元形式中，例如其外壳适合于在肠的下部中释放出内含物的胶囊。在另一个实施方式中，所述球丸可以被包含在胶囊中，所述胶囊本身被包含在包衣胶囊中。在另一个实施方式中，所述球丸可以被包含或包埋在多单元粒子系统中。
卡拉胶是从红色海藻提取的一类天然存在的直链硫酸化多糖。卡拉胶是由重复的硫酸化和非硫酸化的半乳糖和3,6-脱水半乳糖(3,6-AG)单元构成的高分子量多糖。所述单元通过交替的α1-3和β1-4糖苷键相连。三种基本类型的卡拉胶是可商购的，即κ、ι和λ-卡拉胶，它们的区别在于半乳糖单元上的硫酸酯基团的数目和位置。用于本发明的卡拉胶可以选自κ、ι和λ-卡拉胶及其混合物。在这个实施方式的一种情况下，所述吸附剂与κ-卡拉胶混合。在特定实施方式中，所述混合物包含活性炭和κ-卡拉胶。优选地，卡拉胶的量在所述吸附剂和卡拉胶的总重量的约5％至约25％之间，更优选地约10％至约20％之间。在其他特定实施方式中，所述混合物中吸附剂(特别是活性炭)的量在所述吸附剂和卡拉胶的总重量的约95％至约75％之间，更优选地约90％至约80％之间。根据本发明的特定实施方式，卡拉胶的量为所述吸附剂和卡拉胶的总重量的约15％。例如，所述混合物可以含有85％的吸附剂和15％的卡拉胶。
根据本发明的特定实施方式，提供了具有上文指明的重量比的活性炭和卡拉胶、特别是κ-卡拉胶的混合物。
所述核心(或球丸)可以通过专业技术人员已知的任何适合的手段来生产。具体来说，对成粒技术进行改变以适应于生产所述核心。例如，所述核心可以如下获得：将所述吸附剂与卡拉胶以上文指明的比率混合，添加溶剂例如水以进行湿法成粒，然后挤出，任选地然后用滚刀进行滚圆或成球或进行一锅法成球。任何剩余的水可以例如通过使用常规技术干燥得到的球丸来去除。
在一个实施方式中，所述核心或球丸具有50μm至6000μm、特别是100μm至5000μm、特别是150μm至4000μm、特别是250至3000μm、特别是250至1000μm、特别是300至3000μm(例如500至3000μm)、特别是300至1000μm、特别是500至1000μm、特别是500至700μm范围内的平均粒径。
所述核心组合物还可以包含常规赋形剂例如抗粘附剂、粘合剂、填充剂、稀释剂、调味剂、着色剂、润滑剂、助流剂、防腐剂、吸附剂和/或甜味剂。此类赋形剂的量可以变化，但通常在所述球丸重量的0.1至50％的范围内。
正如上文讨论的，本发明的优选制剂包含含有吸附剂并可能增补有卡拉胶的核心，所述核心被包衣层覆盖，使得所述吸附剂在肠的下部中，即在末端回肠、盲肠和/或结肠中释放。
就此而言，在优选实施方式中，所述吸附剂被用作包含下述组分的制剂：
-含有所述吸附剂的核心，和
-外部包衣层，所述外部包衣层围绕所述核心形成，使得所述吸附剂在肠的下部中从所述制剂释放。
在优选实施方式中，所述吸附剂被用作包含下述组分的制剂：
-含有所述吸附剂和卡拉胶的核心，和
-外部包衣层，所述外部包衣层围绕所述核心形成，使得所述吸附剂在肠的下部中从所述制剂释放。
适合的包衣的实例包括pH依赖性肠溶聚合物、偶氮类聚合物、二硫化物聚合物和多糖特别是直链淀粉、果胶(例如用二价阳离子偶联的果胶例如果胶钙或果胶锌)、硫酸软骨素和瓜尔胶。代表性的pH依赖性肠溶聚合物包括偏苯三酸乙酸纤维素(CAT)、邻苯二甲酸乙酸纤维素(CAP)、丙烯酸类聚合物、甲基丙烯酸类聚合物、基于丙烯酸甲酯、甲基丙烯酸甲酯和甲基丙烯酸的阴离子共聚物、邻苯二甲酸羟丙基甲基纤维素(HPMCP)、琥珀酸乙酸羟丙基甲基纤维素(HPMCAS)、甲基丙烯酸和丙烯酸乙酯的共聚物、摩尔比为1：1的甲基丙烯酸和甲基丙烯酸甲酯的共聚物、摩尔比为1：2的甲基丙烯酸和甲基丙烯酸甲酯的共聚物、聚乙烯乙酸邻苯二甲酸酯(PVAP)和虫胶树脂。特别优选的聚合物包括虫胶，基于丙烯酸甲酯、甲基丙烯酸甲酯和甲基丙烯酸的阴离子共聚物例如摩尔比为7：3：1的聚(丙烯酸甲酯-共-甲基丙烯酸甲酯-共-甲基丙烯酸)，以及摩尔比为1：2的甲基丙烯酸和甲基丙烯酸甲酯的共聚物。理想情况下，所述聚合物在等于6.0和更高、优选地6.5和更高的pH下溶解。适合的包衣也可以通过将上文提到的聚合物和共聚物混合来获得。在另一个实施方式中，适合的包衣是时间依赖性包衣或基于时间依赖性聚合物例如乙基纤维素聚合物与藻酸钠的混合物。
在特定实施方式中，所述制剂包含位于所述核心与所述外部pH依赖性层之间的另一个中间包衣。所述中间包衣可以由各种不同的聚合物形成，包括pH依赖性聚合物、pH不依赖性水溶性聚合物、pH不依赖性不溶性聚合物及其混合物。此类pH依赖性聚合物的实例包括虫胶类型的聚合物、基于丙烯酸甲酯、甲基丙烯酸甲酯和甲基丙烯酸的阴离子共聚物、甲基丙烯酸和丙烯酸乙酯的共聚物、邻苯二甲酸羟丙基甲基纤维素(HPMCP)和琥珀酸乙酸羟丙基甲基纤维素(HPMCAS)。pH不依赖性水溶性聚合物的实例包括PVP或高分子量纤维素聚合物例如羟丙基甲基纤维素(HPMC)或羟丙基纤维素(HPC)。pH不依赖性不溶性聚合物的实例包括乙基纤维素聚合物或丙烯酸乙酯和甲基丙烯酸甲酯的共聚物。
在特定实施方式中，本发明使用包含下述组分的制剂：
-核心，其包含吸附剂(优选为活性炭)和卡拉胶(优选为κ-卡拉胶)的混合物，
-中间包衣，其选自HPMC、乙基纤维素以及甲基丙烯酸和丙烯酸乙酯的共聚物例如
Figure BDA0002921556420000101
L30D-55与丙烯酸乙酯和甲基丙烯酸甲酯的共聚物例如
Figure BDA0002921556420000102
NE30D的混合物(例如重量比为1:9至9:1、优选为2:8至3:7的混合物)，和
-外层，其由基于丙烯酸甲酯、甲基丙烯酸甲酯和甲基丙烯酸的阴离子共聚物例如7:3:1的聚(丙烯酸甲酯-共-甲基丙烯酸甲酯-共-甲基丙烯酸)例如
Figure BDA0002921556420000103
FS30D制成。
在特定实施方式中，所述制剂包含含有约85％的活性炭和约15％的κ-卡拉胶的核心以及包衣，所述包衣具有基于丙烯酸甲酯、甲基丙烯酸甲酯和甲基丙烯酸的阴离子共聚物(例如7:3:1的聚(丙烯酸甲酯-共-甲基丙烯酸甲酯-共-甲基丙烯酸)，例如
Figure BDA0002921556420000104
FS30D,Evonik,Darmstadt,Germany)或甲基丙烯酸和丙烯酸乙酯的共聚物(例如
Figure BDA0002921556420000105
L30D55,Evonik,Darmstadt,Germany)的混合物。
在另一个实施方式中，所述吸附剂被配制成如WO2014044794中所公开的组合物，其包含：
(a)包含活性碳的核心；
(b)围绕所述核心的第一层，所述第一层包含不可溶的半透性材料；和
(c)围绕所述第一层的第二层，其在预定pH下溶解或在胃肠道中的预定位置处溶解。
在这个实施方式的变化形式中，所述核心是活性碳。在另一个变化形式中，所述活性碳被喷砂处理或修边。在又一个变化形式中，所述活性碳具有0.02至5.0mm的粒径，例如0.6至1.2mm的粒径。在另一个变化形式中，所述不可溶的半透性材料包含乙基纤维素、单硬脂酸甘油酯、丁酸乙酸纤维素、二聚乳酸、聚氯乙烯和聚(甲基)丙烯酸酯聚合物例如Eudragit RL 100、Eudragit RL PO、Eudragit RL 30D、Eudragit RL 12.5、Eudragit RS100、Eudragit RS PO、Eudragit RS 30D、Eudragit RS 12.5和Eudragit NE 30D、EudragitHE 40D中的一者或多者。在另一个变化形式中，所述第一层还包含水溶性材料，其中所述第一层还可以包含含有羟丙基甲基纤维素(HPMC)的水溶性材料。在某些实施方式中，所述水溶性材料可以与所述不可溶的半透性材料混合，和/或可以包含以重量计0.1至30％的量的所述不可溶的半透性材料，例如以重量计2至25％的量的所述不可溶的半透性材料。在另一个特定变化形式中，所述第一层允许分子通过所述半透膜朝向核心逐渐扩散，与所述活性碳发生接触。在又一个变化形式中，所述第二层包含在pH 5至pH 7下溶解的材料。在某些变化形式中，所述第二层是肠溶层，包含在pH 1至4.9下基本上保持完整但在pH 5至7下快速分解的材料。在一个变化形式中，所述第二层包含pH敏感性聚合物。代表性的第二层包括选自下述的层：琥珀酸乙酸羟丙基甲基纤维素，偏苯三酸乙酸纤维素(CAT)，邻苯二甲酸乙酸纤维素(CAP)，基于丙烯酸甲酯、甲基丙烯酸甲酯和甲基丙烯酸的阴离子共聚物，邻苯二甲酸羟丙基甲基纤维素(HP CP)，琥珀酸乙酸羟丙基甲基纤维素(HPMCAS)，甲基丙烯酸和丙烯酸乙酯的共聚物，甲基丙烯酸和甲基丙烯酸甲酯的共聚物(1:1摩尔比)，甲基丙烯酸和甲基丙烯酸甲酯的共聚物(1:2摩尔比)，聚乙烯乙酸邻苯二甲酸酯(PVAP)和虫胶树脂。在这个实施方式的另一个特定变化形式中，所述活性碳是唯一的活性药物成分。在又一个变化形式中，所述组合物包含：
(a)包含活性碳的核心；
(b)围绕所述核心的第一层，所述第一层包含采取乙基纤维素形式的不可溶的半透性材料，并任选地还包含含有羟丙基甲基纤维素(HPMC)的水溶性材料；和
(c)第二层，其包含琥珀酸乙酸羟丙基甲基纤维素(HPMCAS)。
在另一个变化形式中，所述吸附剂是配制在组合物中的活性碳，所述组合物包含：
(a)核心，其是活性碳；
(b)围绕所述核心的第一层，所述第一层包含不溶于水的半透性材料，并且还包含含有以所述不可溶的半透性材料的重量计2-25％的量的羟丙基甲基纤维素的水溶性材料；和
(c)围绕所述第一层的第二层，其在pH 5至7下溶解。
抗生素
术语“抗生素”是指具有对抗细菌的活性的任何化合物。多亏本发明，可以被消除的抗生素包括但不限于：
-β-内酰胺类，包括：
-青霉素类(例如青霉素G、青霉素V、氨苄青霉素、阿莫西林、巴氨西林、羧苄青霉素、卡茚西林、替卡西林、阿洛西林、美洛西林、哌拉西林等)，
-耐青霉素酶的青霉素类(例如甲氧西林、苯唑西林、氯唑西林、双氯西林、萘夫西林等)，
-头孢菌素类，例如：第一代头孢菌素类(例如头孢羟氨苄、头孢氨苄、头孢拉定、头孢噻吩、头孢匹林、头孢唑啉等)；第二代头孢菌素类(例如头孢克洛、头孢孟多、头孢尼西、头孢西丁、头孢替坦、头孢呋辛、头孢呋辛酯、头孢美唑、头孢丙烯、氯碳头孢、头孢雷特等)；第三代头孢菌素类(例如头孢吡肟、头孢哌酮、头孢噻肟、头孢唑肟、头孢曲松、头孢他啶、头孢克肟、头孢泊肟、头孢布烯等)；第四代头孢菌素类(例如头孢克定、头孢吡肟、头孢唑兰、头孢匹罗、头孢喹诺等)；第五代和其他代头孢菌素类(例如头孢吡普、头孢洛林、头孢唑烷等)，
-碳青霉烯类(例如亚胺培南、美罗培南、厄他培南、多尼培南等)，
-单环内酰胺类(例如氨曲南等)，
-喹诺酮类(例如萘啶酮酸)和氟喹诺酮类(例如西诺沙星、环丙沙星、莫西沙星、左氧氟沙星、氧氟沙星、加替沙星、吉米沙星、诺氟沙星等)，
-磺胺类(例如磺胺、磺胺嘧啶、磺胺甲噁唑、磺胺异噁唑、磺乙酰胺、磺胺对甲氧嘧啶等)，
-氨基糖苷类(例如链霉素、庆大霉素、妥布霉素、阿米卡星、奈替米星、卡那霉素、新霉素B、C和E、壮观霉素、嘌呤霉素、庆大霉素等)，
-四环素类(例如四环素、金霉素、土霉素、美他环素、多西环素、米诺环素、替加环素、艾拉环素等)，
-大环内酯类(例如红霉素、阿奇霉素、克拉霉素、非达霉素、泰利霉素、交沙霉素、竹桃霉素、螺旋霉素、泰乐菌素、罗红霉素、喹红霉素、索利霉素等)，
-糖肽类(例如万古霉素、奥利万星、替拉万星、替考拉宁、达巴万星、雷莫拉宁等)，
-噁唑烷酮类(例如利奈唑胺、泼斯唑来、特地唑胺、雷得唑来、环丝氨酸等)，
-氯霉素类(例如氯霉素、甲砜霉素等)，
-林可酰胺类(例如克林霉素、林可霉素等)，
-链阳菌素类(例如普那霉素、奎奴普丁/达福普丁、维吉霉素等)，
-多粘菌素类(例如多粘菌素A、B、C、D、E1(粘杆菌素A)或E2、粘杆菌素B或C等)，
-二氨基嘧啶类(例如甲氧苄啶(通常与磺胺甲噁唑联合使用)、吡嗪酰胺等)，
-砜类(例如氨苯砜、阿地砜钠等)，
-对氨基苯甲酸，
-杆菌肽，
-异烟胼，
-利福霉素类(例如利福平、利福布丁、利福喷丁、利福拉齐、rimamixin等)，
-乙胺丁醇，
-乙硫异烟胺，
-卷曲霉素，
-氯法齐明，和
-任何其他抗细菌剂。
术语“抗生素”也覆盖抗生素的组合。
抗生素失活酶和酶制剂
在某些实施方式中，本发明使用抗生素失活酶在需要的对象中提高抗癌药剂的效能，其中所述对象已接受、接受或将接受用于预防或治疗感染的抗生素。
在本发明的情形中，“抗生素失活酶”是能够将抗生素水解或失活，从而使所述抗生素无生物学活性的酶。例如，与没有抗生素失活酶的情况下获得的抗生素的最低抑制浓度(MIC)相比，抗生素失活酶可以实质性提高抗生素的MIC。根据本发明，如果对某种浓度的给定抗生素敏感的细菌在用所述失活酶处理后的所述浓度的抗生素存在下的生长与在不存在所述抗生素的情况下的生长相一致，则抗生素失活是完全的。全失活的另一个定义是在用所述失活酶处理后，抗生素对敏感细菌的MIC提高至少2个数量级的情况。
根据本发明所述使用的抗生素失活酶可以是天然的、化学修饰的、遗传工程改造的或合成的。
抗生素失活酶还包括亲代抗生素失活酶的功能性变体，例如β-内酰胺酶、红霉素酯酶和酮还原酶的功能性变体。在本发明的情形中，酶的“功能性变体”是源自于亲代酶的酶，其具有相同类型的催化活性(例如，β-内酰胺酶变体是具有β-内酰胺酶活性的酶)，但具有不同的氨基酸序列。这种功能性变体可能与所述亲本酶具有至少70％、75％、80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、98.5％、99％、99.1％、99.2％、99.3％、99.4％、99.5％、99.6％、99.7％、99.8％、至少99.9％的一致性。这种功能性变体也可能对给定抗生素，例如在β-内酰胺酶的情况下对给定的β-内酰胺类抗生素具有相对于所述亲代抗生素失活酶的比活性来说至少50％、55％、60％、65％、70％、75％、80％、85％、90％、95％、100％、105％、110％、115％、120％、125％、130％、135％、140％、145％、150％、160％、170％、180％、190％、200％、220％、240％、260％、280％、300％、350％、400％、500％、600％、700％、800％或甚至至少1600％的比活性。
可用于本发明的实践的代表性抗生素失活酶包括但不限于使β-内酰胺类抗生素失活的酶(例如β-内酰胺酶)、使氟喹诺酮失活的酶(例如氨基糖苷N-乙酰转移酶)、使大环内酯失活的酶(例如红霉素酯酶或红霉素磷酸转移酶)、使四环素失活的酶(例如NADPH依赖性四环素氧化还原酶)或使林可酰胺失活的酶(例如林可霉素核苷酸转移酶)。
β-内酰胺酶是具有β-内酰胺酶活性的酶(EC 3.5.2.6)，即催化在诸如β-内酰胺类抗生素(例如青霉素类、头孢菌素类、碳青霉烯类、培南砜类)的化合物中存在的β-内酰胺环的酰胺键的不可逆水解，以产生没有其抗细菌活性的水解分子的酶。这类酶对于本领域技术人员来说是公知的(Wang等，1999,Curr Opin Chem Biol.3(5),614-22；Frère,J.M.1995,Mol Microbiol.16(3):385-95)。
在特定实施方式中，所述β-内酰胺酶是丝氨酸β-内酰胺酶或锌依赖性β-内酰胺酶、也被称为金属β-内酰胺酶。在另一个实施方式中，所述β-内酰胺酶选自A类、B类、C类和D类β-内酰胺酶。在其他特定实施方式中，所述β-内酰胺酶选自第1组、第2组、第3组和第4组β-内酰胺酶(Bush等，Antimicrob.Agents Chemother,39:1211)。在某些实施方式中，所述β-内酰胺酶是由来自于地衣芽孢杆菌(Bacillus lichenoformis)749/C的β-内酰胺酶的衍生物组成的P1A、P3A或P4A及其衍生物或来自于蜡样芽孢杆菌(Bacillus cereus)的金属β-内酰胺酶的P2A及其衍生物中的一者或多者。此外，所述β-内酰胺酶可以是广谱β-内酰胺酶(ESBL)，其任选地选自TEM、SHV、CTX-M、OXA、PER、VEB、GES和IBCβ-内酰胺酶。此外，所述β-内酰胺酶可以是抑制剂抗性的β-内酰胺酶，其任选地选自AmpC型β-内酰胺酶、碳青霉烯酶例如但不限于IMP型碳青霉烯酶(金属β-内酰胺酶)、VIM(维罗纳整合子编码的金属β-内酰胺酶)碳青霉烯酶、OXA(苯唑西林酶)组的β-内酰胺酶、KPC(肺炎克雷伯氏菌(K.pneumonia)碳青霉烯酶)、CMY(C类)、SME、IMI、NMC和CcrA以及NDM(新德里金属β-内酰胺酶例如NDM-1)β-内酰胺酶。
在某些实施方式中，所述β-内酰胺酶是VIM(维罗纳整合子编码的金属β-内酰胺酶)。说明性的VIM酶包括但不限于VIM-1、VIM-2、VIM-3、VIM-4和VIM-19。另外的VIM酶描述在例如Queenan等，(2007)Clin.Microbiol.Rev.20(3):440-458中。在其他特定实施方式中，所述β-内酰胺酶是VIM-2或其变体。此类β-内酰胺酶公开在PCT/EP2017/053985、PCT/EP2017/053986和EP17198414中。在特定情况下，本发明涉及使用PCT/EP2017/053985、PCT/EP2017/053986和EP17198414中公开的任何特定实施方式，包括其中公开的任何特定变体VIM-2。在特定实施方式中，所述抗生素失活酶是例如在SEQ ID NO：1中所表示的VIM-2。在另一个特定实施方式中，所述抗生素失活酶是具有如SEQ ID NO：2至46中所示的氨基酸序列的VIM-2功能性变体。在特定实施方式中，所述VIM-2功能性变体具有包含SEQ ID NO：29、SEQ ID NO：31、SEQ ID NO：34或SEQ ID NO：36或由其构成的序列。
在另一个实施方式中，所述β-内酰胺酶是来自于地衣芽孢杆菌(Bacilluslichenoformis)749/C的β-内酰胺酶或其变体，例如P1A、P3A(也被称为“ribaxamase”)或P4A。P1A具有SEQ ID NO：47中示出的序列。
在某些实施方式中，所述β-内酰胺酶是正如在例如WO2007147945中所述的来自于蜡样芽孢杆菌(Bacillus cereus)的金属β-内酰胺酶(也被称为P2A)或其功能性变体。在特定实施方式中，所述P2A酶具有SEQ ID NO：48中示出的序列。所述P2A酶的功能性变体可以与SEQ ID NO：48中示出的序列具有至少约60％、至少约70％、至少约80％、至少约90％、至少约95％、至少约97％、至少约98％、至少约99％或100％的一致性。
在某些实施方式中，所述β-内酰胺酶是正如在例如WO2011148041中所述的P3A或其功能性变体。在特定实施方式中，所述P3A酶具有SEQ ID NO：49(所述酶的成熟形式)或SEQ ID NO：50(所述酶的包括31个氨基酸长的信号肽的形式)中示出的序列。所述P3A酶的功能性变体可以与SEQ ID NO：49或SEQ ID NO：50中示出的序列具有至少约60％、至少约70％、至少约80％、至少约90％、至少约95％、至少约97％、至少约98％、至少约99％或100％的一致性。在特定实施方式中，所述β-内酰胺酶包含与SEQ ID NO：49具有至少80％序列一致性的氨基酸序列，并且特征在于它在对应于根据Ambler分类的第276位的位置处具有天冬氨酸(D)之外的亲水性氨基酸残基，并且所述亲水性氨基酸选自精氨酸(R)、组氨酸(H)、赖氨酸(K)、天冬酰胺(N)、谷氨酰胺(Q)、丝氨酸(S)和苏氨酸(T)。在其他特定实施方式中，所述β-内酰胺酶包含与SEQ ID NO：49具有至少80％序列一致性的氨基酸序列，并且特征在于它在对应于根据Ambler分类的第276位的位置处具有天冬酰胺(N)。在又一个实施方式中，所述β-内酰胺酶具有SEQ ID NO：49中示出的氨基酸序列，其中在对应于根据Ambler分类的第276位的位置处的氨基酸残基是天冬酰胺(N)。
在另一个实施方式中，所述β-内酰胺酶是正如在例如WO2015/161243中所述的P4A或其功能性变体。在特定实施方式中，所述P4A酶具有SEQ ID NO：79或SEQ ID NO：80的序列。所述P4A酶的功能性变体可以与SEQ ID NO：79或SEQ ID NO：80中示出的序列具有至少约60％、至少约70％、至少约80％、至少约90％、至少约95％、至少约97％、至少约98％、至少约99％或100％的一致性。
在某些实施方式中，所述β-内酰胺酶是肺炎克雷伯氏菌(Klebsiellapneumoniae)碳青霉烯酶(KPC)。说明性的KPC包括但不限于KPC-1/2(SEQ ID NO：51)、KPC-3(SEQ ID NO：52)、KPC-4(SEQ ID NO：53)、KPC-5(SEQ ID NO：54)、KPC-6(SEQ ID NO：55)、KPC-7(SEQ ID NO：56)、KPC-8(SEQ ID NO：57)、KPC-9(SEQ ID NO：58)、KPC-10(SEQ ID NO：59)、KPC-11(SEQ ID NO：60)、KPC-12(SEQ ID NO：61)、KPC-13(SEQ ID NO：62)、KPC-14(SEQID NO：63)、KPC-15(SEQ ID NO：64)和KPC-17(SEQ ID NO：65)。在一个实施方式中，所述β-内酰胺酶是KPC-1/2。在一个实施方式中，所述β-内酰胺酶是KPC-3。KPC酶的功能性变体可以与SEQ ID NO：51至SEQ ID NO：65中示出的序列具有至少约60％、至少约70％、至少约80％、至少约90％、至少约95％、至少约97％、至少约98％、至少约99％或100％的一致性。
在另一个实施方式中，所述β-内酰胺酶是新德里金属β-内酰胺酶(NDM)。说明性的NDM包括但不限于NDM-1(SEQ ID NO：66)、NDM-2(SEQ ID NO：67)、NDM-3(SEQ ID NO：68)、NDM-4(SEQ ID NO：69)、NDM-5(SEQ ID NO：70)、NDM-6(SEQ ID NO：71)、NDM-7(SEQ ID NO：72)、NDM-8(SEQ ID NO：73)、NDM-9(SEQ ID NO：74)、NDM-10(SEQ ID NO：75)、NDM-11(SEQID NO：76)、NDM-12(SEQ ID NO：77)和NDM-13(SEQ ID NO：78)。在一个实施方式中，所述β-内酰胺酶是NDM-1。在一个实施方式中，所述广谱碳青霉烯酶是NDM-4。NDM酶的功能性变体可以与SEQ ID NO：66至SEQ ID NO：78中示出的序列具有至少约60％、至少约70％、至少约80％、至少约90％、至少约95％、至少约97％、至少约98％、至少约99％或100％的一致性。
在某些实施方式中，所述β-内酰胺酶是IMP型碳青霉烯酶。说明性的IMP型酶包括但不限于IMP-1、IMP-4、IMP-8、IMP-11、IMP-43和IMP-44。另外的IMP型酶描述在例如Queenan等，(2007)Clin.Microbiol.Rev.20(3):440-458中。
在某些实施方式中，所述β-内酰胺酶来自于OXA(苯唑西林酶)组的β-内酰胺酶。说明性的OXAβ-内酰胺酶包括但不限于OXA-23、OXA-24、OXA-27、OXA-40、OXA-48、OXA-49、OXA-50、OXA-51、OXA-58、OXA-64、OXA-71和OXA-181。另外的OXA型β-内酰胺酶描述在例如Walther-Rasmussen等，Journal of Antimicrobial Chemotherapy(2006),57:373-383和Queenan等，(2007)Clin.Microbiol.Rev.20(3):440-458中。
在某些实施方式中，所述β-内酰胺酶是CMY(C类碳青霉烯酶)酶。说明性的具有碳青霉烯酶活性的CMY酶是CMY-10，正如在例如Lee等，(2006)Research Journal ofMicrobiology(1):1-22中所述。
在某些实施方式中，所述β-内酰胺酶是SME酶(表示粘质沙雷氏菌(Seiratiamarcescens))。说明性的SME酶包括但不限于SME-1、SME-2或SME-3，正如在例如Queenan等，(2007)Clin.Microbiol.Rev.20(3):440-458中所述。
在某些实施方式中，所述β-内酰胺酶是IMI酶(水解亚胺培南的β-内酰胺酶)。说明性的IMI酶包括但不限于IMI-1或IMI-2，正如在例如Queenan等，(2007)Clin.Microbiol.Rev.20(3):440-458中所述。
在某些实施方式中，所述β-内酰胺酶是NMC酶(不是金属酶的碳青霉烯酶)。说明性的NMC酶是NMC-A，正如在例如Queenan等，(2007)Clin.Microbiol.Rev.20(3):440-458中所述。
在某些实施方式中，所述β-内酰胺酶是GES酶(圭亚那广谱)。说明性的GES酶包括但不限于GE-2、GES-4、GES-5、GES-6、GES-7、GES-8、GES-9、GES-11、GES-14和GES-18，正如在例如Queenan等，(2007)Clin.Microbiol.Rev.20(3):440-458和Johnson等，(2014)A、B和D类β-内酰胺酶的晶体结构(Crystal Structures of Class A,B,and Dβ-Lactamases)(http://www.carbapenemase.ca/crystal_structures.html)中所述。
在某些实施方式中，所述β-内酰胺酶是来自于脆弱拟杆菌(Bacteroidesfragilis)的CcrA(CfiA)金属β-内酰胺酶。
在某些实施方式中，所述β-内酰胺酶是来自于居泉沙雷氏菌(Serratiafonticola)的SFC-1酶或来自于肺炎克雷伯氏菌(Klebsiella pneumoniae)的SHV-38酶，正如在例如Walther-Rasmussen等，(2007)Journal of Antimicrobial Chemotherapy,60:470-482中所述。
在另一个实施方式中，所述抗生素失活酶是红霉素酯酶。红霉素酯酶(EC编号3.1.1)是指催化红霉素以及其他大环内酯类抗生素的失活的一类酶。这些酶水解大环内酯类例如红霉素和竹桃霉素的内酯环，正如在Barthelemy等，1984,J.Antibiot.37,1692-1696中所解释的。已知的红霉素酯酶是细菌起源的。它们由例如大肠埃希氏杆菌(Escherichia coli)、Halobacterium salinarum、Gramella forsetii、反硝化无色杆菌(Achromobacter denitrificans)或红球菌属(Rhodococcus sp.)产生。在特定实施方式中，所述红霉素酯酶是通常由对红霉素具有高度抗性的肠杆菌科的成员产生的酶之一，正如在Arthur等，1987,Antimicrob.Agents Chemother.31(3),404-409中所述。两种来自于大肠杆菌的红霉素酯酶已被记录在参考名EreA和EreB下，在本发明中设想了这两者的使用。在本发明的特定实施方式中，所述红霉素酯酶是来自于大肠杆菌的EreB红霉素酯酶(参见Arthur等，1986,Nucleic Acids Res 14(12),4987-4999)。
在另一个实施方式中，所述抗生素失活酶是酮还原酶。酮还原酶(KRED)或羰基还原酶类(EC 1.1.1.184)酶可用于从相应的前手性酮底物合成光学活性的醇类。KRED通常将酮底物转变成相应的醇产物，但是也可以催化逆反应，将醇底物氧化成相应的酮/醛产物。
在另一个实施方式中，所述抗生素失活酶是杂合蛋白分子。代表性的杂合蛋白分子是在美国专利申请20170354706中公开的那些。此类杂合蛋白分子可以包含键合在一起的两种酶，能够使至少一种抗生素失活。在特定实施方式中，这些酶被合并成单个单链蛋白。这两种酶可以均来自于相同类别，或各自来自于不同类别。例如，所述两种酶可以是β-内酰胺酶，或在β-内酰胺酶、使氨基糖苷失活的酶、使氟喹诺酮失活的酶、使林可酰胺失活的酶、使大环内酯失活的酶或使四环素失活的酶的类别中选择。在特定实施方式中，所述杂合蛋白分子中的每种酶使不同抗生素失活。在另一个实施方式中，所述杂合蛋白分子包含能够使属于相同类别的抗生素失活的两种酶。在特定实施方式中，所述杂合蛋白中的至少一种组分酶的序列与SEQ ID NO：81至SEQ ID NO：87具有至少40％、45％、50％、55％、60％、65％、70％、75％、80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、98.5％、99％、99.1％、99.2％、99.3％、99.4％、99.5％、99.6％、99.7％、99.8％或至少99.9％的序列同源性。在其他特定实施方式中，所述杂合蛋白中的至少一种组分酶的序列具有由SEQID NO：81、SEQ ID NO：82、SEQ ID NO：83、SEQ ID NO：84、SEQ ID NO：85、SEQ ID NO：86或SEQ ID NO：87构成的序列。在其他特定实施方式中，所述杂合蛋白分子具有包含选自SEQID NO：88至90的序列或由其构成的氨基酸序列。
在另一个实施方式中，所述酶不论是生物还是合成生产的，均可以被进一步酶法和/或化学修饰，以便增强其活性、稳定性、溶解性或任何其他有益特征。这种修饰的一个实例是将聚乙二醇连接到表面氨基或PEG化。
在特定实施方式中，所述抗生素失活酶被配制成适合于在肠的所需部分中释放出所述酶的制剂。在特定实施方式中，所述肠的所需部分是肠的下部，例如回肠、盲肠或结肠。在另一个特定实施方式中，所述肠的所需部分是肠的上部，例如十二指肠或空肠。在特定实施方式中，所述制剂包含用肠溶包衣(例如用在高于或等于7.0的pH下溶解的肠溶包衣)包被的酶的球丸。在另一个特定实施方式中，所述制剂包含在肠溶包衣胶囊(例如具有在高于或等于5.5的pH或高于或等于7.0的pH下溶解的肠溶包衣)内的肠溶包衣的酶球丸(例如具有在高于或等于5.5的pH或高于或等于7.0的pH下溶解的肠溶包衣)。在另一个特定实施方式中，所述制剂包含在未包衣的胶囊中的肠溶包衣的球丸。所述制剂的选择可以取决于所述抗生素向所述对象的给药途径。例如，在所述抗生素肠胃外给药的情况下，可以考虑在肠的上部或下部释放所述抗生素失活酶的制剂。在所述抗生素口服给药的情况下，所述酶制剂优选地在肠的下部中，在所述抗生素被所述酶的失活不再能够干扰所述抗生素在小肠中的所需吸收的位置处释放出所述酶，以便从所述抗生素的正面效应获益。
在特定实施方式中，所述制剂包括消化道蛋白酶例如胃蛋白酶、胰蛋白酶、胰凝乳蛋白酶、羧肽酶、弹性蛋白酶的抑制剂，以便保护所述酶并延长其在肠道中有活性的时间。
使用方法
本发明涉及如上所提供的吸附剂，其用于提高抗癌药剂例如免疫肿瘤药剂的治疗效能的方法中。本发明还涉及如上所提供的吸附剂，其用于与抗癌药剂例如免疫肿瘤药剂相组合治疗或预防癌症的方法中。本发明还涉及如上所提供的吸附剂，其用于与抗癌药剂例如免疫肿瘤药剂相组合治疗或预防癌症的方法中，从而提高所述抗癌药剂的效能。本发明还涉及如上所提供的吸附剂，其用于与抗癌药剂例如免疫肿瘤药剂相组合治疗或预防癌症的方法中，从而保留所述抗癌药剂的效能。本发明还涉及如上所提供的吸附剂，其用于与抗癌药剂例如免疫肿瘤药剂相组合治疗或预防癌症的方法中，从而增强所述抗癌药剂的效能。
所述吸附剂可以在所述疗法中的任何时点，例如在所述抗癌药剂例如免疫肿瘤药剂之前、期间和/或之后给药。具体来说，所述吸附剂可以在所述患者一被诊断为患有恶性肿瘤后就立即给药，即使给药抗癌药剂的打算仅占极少的可能性。有时也被称为抗肿瘤药剂的抗癌药剂是在哺乳动物例如人类中用于对抗癌症的物质。术语“抗癌药剂”包括但不限于直接影响癌细胞或例如通过影响血管化间接影响癌细胞的化学品和生物药剂。例如，抗癌药剂包括但不限于化学治疗分子例如细胞生长抑制剂、细胞毒性药剂和抗血管生成药剂、靶向癌细胞的抗癌抗体、抗癌肽和抗癌病毒。说明性的抗癌药剂包括但不限于：
-微管蛋白毒素，紫杉烷，例如多西他赛、紫杉醇，
-铂化合物，例如顺铂、卡铂、奥沙利铂，
-干扰DNA复制的药剂例如DNA嵌入剂，例如蒽环霉素类，
-拓扑异构酶抑制剂，例如依托泊苷，
-抗代谢物，例如甲氨蝶呤、阿糖胞苷(ara-C)、吉西他滨、5-氟尿嘧啶，
-烷化剂，例如氮芥、美法仑、卡莫司汀、异环磷酰胺或环磷酰胺，
-靶向药剂，例如酶抑制剂、特别是激酶抑制剂例如埃罗替尼、索拉非尼、伊马替尼，或蛋白酶体抑制剂例如硼替佐米、卡非佐米、伊沙佐米，
-靶向生长因子受体的细胞外区域的单克隆抗体，例如曲妥珠单抗、贝伐单抗和西妥昔单抗，
-免疫肿瘤药剂，和
-它们的组合。
蒽环霉素类包括但不限于多柔比星和道诺霉素。
拓扑异构酶抑制剂还包括但不限于喜树碱、伊立替康、拓扑替康及其衍生物。
抗代谢物还包括但不限于卡培他滨和培美曲塞。
在特定实施方式中，所述抗癌药剂是免疫肿瘤药剂。免疫肿瘤药剂(也被称为免疫靶向药剂)至少部分通过参与免疫系统或通过免疫系统相关的作用方式针对肿瘤发挥作用。更特别地，免疫肿瘤药剂可以通过调节免疫细胞的行动起作用。
免疫肿瘤药剂的实例包括调节免疫检查点的药剂，例如2B4、4-1BB(CD137)、AaR、B7-H3、B7-H4、BAFFR、BTLA、CD2、CD7、CD27、CD28、CD30、CD40、CD80、CD83配体、CD86、CD160、CD200、CDS、CEACAM、CTLA-4、GITR、HVEM、ICAM-1、KIR、LAG-3、LAIR1、LFA-1(CD 11a/CD 18)、LIGHT、NKG2C、NKp80、OX40、PD-1、PD-L1、PD-L2、SLAMF7、TGFRp、TIGIT、Tim3和VISTA。
免疫肿瘤药剂可以采取抗体、肽、小分子或病毒的形式。在特定实施方式中，所述免疫肿瘤药剂是针对PD-1、PD-L1或PD-L2的抗体。
在特定实施方式中，所述免疫肿瘤药剂是精氨酸酶、CTLA-4、吲哚胺2,3-加双氧酶和/或PD-1/PD-L1的抑制剂。在某些实施方式中，所述免疫肿瘤药剂是阿巴伏单抗、阿德木单抗、阿夫土珠单抗、阿仑单抗、麻安莫单抗、阿泊珠单抗、博纳吐单抗、BMS-936559、卡妥索单抗、德瓦鲁单抗、epacadostat、依帕珠单抗、indoximod、奥英妥珠单抗、intelumumab、伊匹单抗、isatuximab、lambrolizumab、MED 14736、MPDL3280A、纳武单抗、obinutuzumab、ocaratuzumab、奥法木单抗、olatatumab、派姆单抗、pidilizumab、利妥昔单抗、ticilimumab、samalizumab或tremelimumab。
更通常地，免疫肿瘤药剂可以是可用于治疗恶性肿瘤疾病并且至少部分通过参与免疫系统起作用或具有免疫系统相关的作用方式的任何药剂。例如，所述免疫肿瘤药剂可以选自但不限于：
-免疫检查点抑制剂，例如PD-1抑制剂例如纳武单抗或派姆单抗；
-免疫检查点抑制剂，例如PDL-1抑制剂例如阿特珠单抗、阿维单抗或德瓦鲁单抗，或CTLA-4抑制剂例如伊匹单抗，
-癌症疫苗，例如西普鲁塞-T；
-免疫调节剂，例如沙利度胺、来那度胺、泊马度胺；
-非特异性免疫疗法，例如干扰素或白介素；和
-嵌合抗原受体(CAR)-T细胞疗法，例如tisagenlecleucel或axicabtageneciloleucel；以及
-它们的组合。
在特定实施方式中，所述抗癌药剂是抗PD-1抗体。在其他特定实施方式中，所述抗PD-1抗体选自纳武单抗和派姆单抗。
在本发明的特定实施方式中，所述抗癌药剂选自阿法替尼、阿柏西普、阿仑单抗、阿利维甲酸、六甲蜜胺、阿那格雷、三氧化二砷、天冬酰胺酶、阿特珠单抗、阿维单抗、阿西替尼、阿扎胞苷、苯达莫司汀、贝伐单抗、蓓萨罗丁、博来霉素、硼替佐米、博舒替尼、白消安、卡巴他赛、卡培他滨、卡铂、卡莫氟、卡莫司汀、西妥昔单抗、苯丁酸氮芥、盐酸氮芥、顺铂、克拉屈滨、氯法拉滨、克唑替尼、环磷酰胺、阿糖胞苷、达卡巴嗪、放线菌素D、达沙替尼、道诺霉素、地西他滨、地尼白介素、地诺单抗、多西他赛、多柔比星、德瓦鲁单抗、表柔比星、埃罗替尼、雌莫司汀、依托泊苷、依维莫司、氟尿苷、氟达拉滨、氟尿嘧啶、福莫司汀、吉非替尼、吉西他滨、吉妥单抗、羟基脲、替伊莫单抗、伊达比星、异环磷酰胺、伊马替尼、伊匹单抗、伊立替康、异维甲酸、伊沙匹隆、拉帕替尼、来那度胺、洛莫司汀、美法仑、巯基嘌呤、甲氨蝶呤、丝裂霉素、米托蒽醌、奈达铂、奈拉滨、尼罗替尼、纳武单抗、奥法木单抗、奥沙利铂、紫杉醇、帕尼单抗、帕比司他、帕唑帕尼、派姆单抗、培美曲塞、喷司他丁、帕妥珠单抗、泊马度胺、帕纳替尼、丙卡巴肼、雷替曲塞、瑞戈非尼、利妥昔单抗、罗米地辛、鲁索替尼、索拉非尼、链脲佐菌素、舒尼替尼、他米巴罗汀、替加氟、替莫唑胺、西罗莫司脂化物、替尼泊苷、沙利度胺、硫鸟嘌呤、拓扑替康、托西莫单抗、曲妥珠单抗、维甲酸、丙戊酸盐、戊柔比星、凡德他尼、维罗非尼、长春花碱、长春新碱、长春地辛、长春氟宁、长春瑞滨和伏立诺他。
本发明的吸附剂和抗癌药剂可用于在对象中治疗或预防一种癌症或多种癌症。在某些实施方式中，所述癌症可以是选自下述的癌症之一或其变化形式：急性成淋巴细胞性白血病(ALL)，急性髓系白血病(AML)，肾上腺皮质癌，肛门癌，阑尾癌，非典型畸胎瘤/横纹肌样瘤，基底细胞癌，胆管癌，膀胱癌，骨癌，脑肿瘤，星形细胞瘤，脑和脊髓肿瘤，脑干神经胶质瘤，中枢神经系统非典型畸胎瘤/横纹肌样瘤，中枢神经系统胚胎性肿瘤，乳腺癌，支气管肿瘤，伯基特淋巴瘤，类癌瘤，原发灶不明的转移癌，中枢神经系统癌症，宫颈癌，儿童期癌症，脊索瘤，慢性淋巴细胞性白血病(CLL)，慢性髓系白血病(CML)，慢性骨髓增殖性障碍，结肠癌，结肠直肠癌，颅咽管瘤，皮肤T细胞淋巴瘤导管原位癌(DCIS)，胚胎性肿瘤，子宫内膜癌，成室管膜细胞瘤，室管膜瘤，食管癌，嗅神经母细胞瘤，尤文氏肉瘤，颅外生殖细胞瘤，性腺外生殖细胞瘤，肝外胆管癌，眼癌，骨纤维组织细胞瘤，胆囊癌，胃癌，胃肠类癌瘤，胃肠间质瘤(GIST)，生殖细胞瘤，颅外生殖细胞瘤，性腺外生殖细胞瘤，卵巢生殖细胞瘤，妊娠滋养细胞肿瘤，神经胶质瘤，毛细胞白血病，头颈癌，心癌，肝细胞癌，组织细胞增生症，朗格汉斯细胞癌，霍奇金淋巴瘤，下咽癌，眼内黑素瘤，胰岛细胞瘤，卡波斯肉瘤，肾癌，朗格汉斯细胞组织细胞增生症，喉癌，白血病，唇和口腔癌，肝癌，原位小叶癌(LCIS)，肺癌,淋巴瘤，AIDS相关淋巴瘤，巨球蛋白血症，男性乳腺癌，成神经管细胞瘤，髓上皮瘤，黑素瘤，梅克尔细胞癌，恶性间皮瘤，隐匿原发的转移性鳞状颈部癌，涉及NUT基因的中线导管癌，口腔癌，多发性内分泌肿瘤综合征，多发性骨髓瘤/浆细胞瘤，蕈样肉芽肿，骨髓增生异常综合征，骨髓增生异常/骨髓增殖性肿瘤，慢性髓系白血病(CML)，急性髓系白血病(AML)，骨髓瘤，多发性骨髓瘤，慢性骨髓增殖性障碍，鼻腔癌，副鼻窦癌，鼻咽癌，成神经细胞瘤，非霍奇金淋巴瘤，非小细胞肺癌，口癌，口腔癌，唇癌，口咽癌，骨肉瘤，卵巢癌，胰腺癌，乳头状瘤病，副神经节瘤，副鼻窦癌，鼻腔癌，甲状旁腺癌，阴茎癌，咽癌，嗜铬细胞瘤，中间分化的松果体实质肿瘤，松果体母细胞瘤，垂体瘤，浆细胞瘤，胸膜肺母细胞瘤，乳腺癌，原发性中枢神经系统(CNS)淋巴瘤，前列腺癌，直肠癌，肾细胞癌，透明细胞肾细胞癌，肾盂癌，输尿管癌，移行细胞癌，成视网膜细胞瘤，横纹肌肉瘤，唾液腺癌，肉瘤，塞扎里综合征，皮肤癌，小细胞肺癌，小肠癌，软组织肉瘤，鳞状细胞癌，隐匿原发的鳞状颈癌(例如转移性)，头颈部鳞状细胞癌(HNSCC)，胃癌，幕上原始神经外胚层肿瘤，T-细胞淋巴瘤，睾丸癌，喉癌，胸腺瘤，胸腺癌，甲状腺癌，肾盂和输尿管的移行细胞癌，三阴性乳腺癌(TBC)，原发灶不明的妊娠滋养细胞肿瘤，儿童期罕见癌症，尿道癌，子宫癌，子宫肉瘤，华氏巨球蛋白血症和威尔姆氏瘤。
具体来说，所述癌症可以选自：
-上皮起源的影响器官例如乳腺(乳腺腺癌)、皮肤(黑素瘤)、肺(非小细胞肺癌和小细胞肺癌)、肾(肾细胞癌)、胰腺(胰腺癌)、膀胱的肿瘤，
-消化道肿瘤例如胃-食管腺癌，
-头颈癌(特别是鳞状肿瘤)，
-鳞状肺部肿瘤，
-影响血液免疫细胞的恶性肿瘤，例如多发性骨髓瘤、淋巴瘤(所有类型的霍奇金和非霍奇金淋巴瘤)、白血病包括淋巴细胞性白血病(例如急性成淋巴细胞性白血病(ALL)或慢性淋巴细胞性白血病,(CLL))、髓系白血病(例如急性髓系白血病(AML)和慢性髓系白血病(CML))、毛细胞白血病、T-细胞前淋巴细胞性白血病、大颗粒淋巴细胞性白血病、成年T-细胞白血病、成年T-细胞淋巴瘤/白血病。
在特定实施方式中，所述癌症选自肺部癌症、黑素瘤、胰腺癌症、肾脏癌症、难治性白血病和淋巴瘤。
在某些实施方式中，本发明的方法还可以包括与所述抗癌药剂联合给药一种或多种另外的治疗剂。可以与所述抗癌药剂联合给药的代表性治疗剂包括但不限于：氨鲁米特，安吖啶，阿那曲唑，天冬酰胺酶，AZD5363，卡介苗(beg)，比卡鲁胺，博来霉素，硼替佐米，布舍瑞林，白消安，喜树碱，卡培他滨，卡铂，卡非佐米，卡莫司汀，苯丁酸氮芥，氯喹，顺铂，克拉屈滨，氯膦酸盐，考比替尼，秋水仙素，环磷酰胺，环丙孕酮，阿糖胞苷，达卡巴嗪，放线菌素D，道诺霉素，脱甲绿胶酶素，地塞米松，二氯乙酸盐，双烯雌酚，己烯雌酚，多西他赛，多柔比星，表柔比星，埃罗替尼，雌二醇，雌莫司汀，依托泊苷，依维莫司，依西美坦，非格司亭，氟达拉滨，氟氢可的松，氟尿嘧啶，氟甲睾酮，氟他胺，吉西他滨，金雀异黄酮，戈舍瑞林，羟基脲，伊达比星，异环磷酰胺，伊马替尼，干扰素，伊立替康，来那度胺，来曲唑，亚叶酸，亮丙瑞林，左旋咪唑，洛莫司汀，氯尼达明，氮芥，甲羟孕酮，甲地孕酮，美法仑，巯基嘌呤，美司钠，甲福明，甲氨蝶呤，米替福新，丝裂霉素，米托坦，米托蒽醌，MK-2206，尼鲁米特，诺考达唑，奥曲肽，奥拉帕尼，奥沙利铂，紫杉醇，帕米磷酸盐，帕唑帕尼，喷司他丁，哌立福辛，普卡霉素，泊马度胺，卟吩姆钠，丙卡巴肼，雷替曲塞，利妥昔单抗，卢卡帕尼，司美替尼，索拉非尼，链脲佐菌素，舒尼替尼，苏拉明，他唑帕尼，他莫昔芬，替莫唑胺，西罗莫司脂化物，替尼泊苷，睾酮，沙利度胺，硫鸟嘌呤，噻替哌，二氯二茂钛，拓扑替康，曲美替尼，曲妥珠单抗，维甲酸，维利帕尼，长春花碱，长春新碱，长春地辛和长春瑞滨。可以与所述抗癌药剂联合给药的其他代表性治疗剂包括但不限于培美曲塞。
在特定实施方式中，抗癌疗法是使用免疫肿瘤药剂和至少一种其他抗癌药剂的组合疗法。例如，所述患者可以用免疫肿瘤药剂和选自铂盐(例如顺铂、卡铂等)、培美曲塞和依托泊苷的至少一种其他抗癌药剂给药。例如所述至少一种其他抗癌药剂可以是：
-培美曲塞，
-培美曲塞和铂盐，
-依托泊苷，或
-依托泊苷和铂盐。
在另一个实施方式中，本发明提供了一种药剂盒，其包含抗癌药剂和吸附剂。在某些实施方式中，所述药剂盒可用于治疗本文中所描述的病症或疾病。
本发明提供了一种治疗或预防癌症的方法，所述方法包括联合给药吸附剂和抗癌药剂。多亏了本发明，给药所述抗癌药剂和吸附剂相对于所述抗癌药剂的单独给药来说提供了提高的效能。
在某些实施方式中，所述抗癌药剂在所述吸附剂后约5分钟之内至约7小时之内给药。在特定实施方式中，所述吸附剂在所述抗癌药剂给药之前多次给药，以便确保所述患者的抗癌免疫监督系统得以改善。例如，所述吸附剂可以每天、每天一次或几次给药共几天。例如，所述吸附剂可以在所述抗癌药剂给药之前1、至少2、至少3、至少4、至少5、至少6或至少7天每天给药一次或几次。
在某些情况下，所述吸附剂用于患有癌症并且除了所述抗癌药剂之外还给药、将给药或已给药可能扰乱所述患者的肠道微生物菌群的物质的对象。多亏了本发明，可以阻止此类物质的有害影响，并因此可以提高所述抗癌药剂的效能。因此，本发明涉及一种减轻物质对患有癌症的对象的肠道微生物菌群可能具有的有害效应的方法，所述对象是抗癌药剂治疗的接受者，所述方法包括向所述对象给药有效量的吸附剂。
在某些实施方式中，所述物质是被给药以治疗所述患者中的病理状况的药物物质。事实上，某些药物物质可以被给药以便治疗疾病，但当它们到达肠的下部时可能对肠道微生物菌群具有有害影响。所述对象仍将接受所述药物物质以从它的所需效应获益，但在另一方面，应该提供避免其次级效应的解决方案。说明性的具有这种行为的物质包括抗生素。抗生素可以被给药到对象以便治疗细菌感染。然而，由于抗生素在设计上能够影响细菌生长或存活，因此它们威胁肠道微生物菌群平衡，并且当它们到达肠的下部时可能引起生态失调。这种诱导的生态失调进而可能导致给药到所述对象的抗癌药物的效能降低。其他说明性的可能引起生态失调的药物物质(也被称为“引起生态失调的药物物质”)包括但不限于：
-化学治疗剂，例如紫杉烷(例如多西他赛、紫杉醇)、蒽环霉素类(例如多柔比星)、拓扑异构酶抑制剂(例如依托泊苷、伊立替康)、抗代谢物(例如甲氨蝶呤、阿糖胞苷、5-氟尿嘧啶、吉西他滨、培美曲塞)、烷化剂(例如美法仑)、激酶抑制剂(例如埃罗替尼)，
-抗真菌剂，例如伏立康唑、两性霉素B脂质体、泊沙康唑，
-抗病毒剂，例如阿昔洛韦、美替沙腙，
-抗炎药剂，例如阿司匹林、布洛芬，以及
-质子泵抑制剂，例如奥美拉唑、泮托拉唑、埃索美拉唑。
因此，在本发明的另一方面，所述吸附剂被给药到患有癌症并正用、将用、已用引起生态失调的药物物质例如抗生素治疗的对象。
同样地，在某些情况下，本文中所描述的抗生素失活酶用于患有癌症并正给药、将给药或已给药抗生素的对象。在这种情况下，所述抗生素失活酶选自能够使给药到所述对象的特定抗生素失活的酶(例如，在所述抗生素是β-内酰胺类抗生素的情况下给药β-内酰胺酶；在另一个实例中，如果所述抗生素是大环内酯类抗生素，则给药红霉素酯酶)。多亏本发明的这个方面，可以阻止所述抗生素的有害影响，并因此可以提高所述抗癌药剂的效能。因此，本发明涉及一种减轻抗生素对患有癌症的对象的肠道微生物菌群可能具有的有害效应的方法，所述对象是抗癌药剂疗法的接受者，所述方法包括向所述对象给药有效量的抗生素失活酶。
所述吸附剂或抗生素失活酶(如果适合的话，由于所述引起生态失调的药物物质是抗生素)甚至可以在所述抗癌药剂开始给药之前很久就给药到所述对象。例如，所述对象可能已被诊断为患有恶性肿瘤，但治疗不能在几天、几周、几个月或几年前开始。在这种情况下，如果所述对象在这些事件之间患有需要使用引起生态失调的药剂例如抗生素治疗的疾病，则通过给药本文中所提供的吸附剂或抗生素失活酶以防止肠道微生物菌群生态失调，将是有利的。同样地，所述吸附剂或抗生素失活酶甚至可以在所述抗癌药剂的给药开始之前或结束之后很长时间给药到所述对象。首先，所述对象的癌症可能不幸复发。在这种情况下，在对象接受引起生态失调的药物物质例如抗生素时停止吸附剂或抗生素失活酶的系统性给药可能严重损害将来使用相同或另一种抗癌药剂的疗法的效能。其次，某些疗法例如基因疗法可能在给药后几年有效，只要所述治疗性基因表达即可。在这种情况下，所述吸附剂或抗生素失活酶的给药对于提高这种长期持续的抗癌疗法是有益的。当然，所述吸附剂或抗生素失活酶优选地在所述抗癌药剂疗法的整个过程中给药，此时所述对象接受使用引起生态失调的药物物质例如抗生素的疗法。
在特定实施方式中，本发明涉及一种用于在需要抗癌药剂的对象中提高这种抗癌药剂的效能的吸附剂，其中所述对象也用引起生态失调的药物物质例如抗生素给药。
在另一个特定实施方式中，本发明涉及一种用于在需要抗癌药剂的对象中提高这种抗癌药剂的效能的抗生素失活酶，其中所述对象也用抗生素给药。
本发明还涉及一种吸附剂，其用于在对象用引起生态失调的药物物质例如抗生素给药时在所述对象中防止抗癌药剂效能的降低。
本发明还涉及一种抗生素失活酶，其用于在对象用抗生素给药时在所述对象中防止抗癌药剂效能的降低。
本发明还涉及一种吸附剂，其用于在对象用引起生态失调的药物物质例如抗生素给药时在所述对象中维持抗癌药剂的效能。
此外，本发明还涉及一种抗生素失活酶，其用于在对象用抗生素给药时在所述对象中维持抗癌药剂的效能。
本发明还涉及一种吸附剂，其与引起生态失调的药物物质例如抗生素一起用于需要抗癌药剂疗法的对象中。
另外，本发明还涉及一种抗生素失活酶，其与抗生素一起用于需要抗癌药剂疗法的对象中。
本发明还涉及一种吸附剂，其与引起生态失调的药物物质例如抗生素相组合，用于治疗或预防可以用所述引起生态失调的药物物质治疗或预防的疾病的方法中，其中需要所述治疗的对象也需要抗癌疗法。
本发明还涉及一种抗生素失活酶，其与抗生素相组合用于治疗或预防可以用所述抗生素治疗或预防的疾病，其中需要所述治疗的对象也需要抗癌疗法。
本发明还涉及一种吸附剂，其用于在需要抗癌药剂的对象中阻止引起生态失调的药物物质例如抗生素对所述抗癌药剂的效能的影响。
另外，本发明涉及一种抗生素失活酶，其用于在需要抗癌药剂的对象中阻止抗生素对所述抗癌药剂的效能的影响。
本发明还涉及一种吸附剂，其用于在需要抗癌药剂的对象中阻止可能由引起生态失调的药物物质例如抗生素引起的所述抗癌药剂效能的降低，所述引起生态失调的药物物质被给药到所述对象是为了治疗或预防可以用所述引起生态失调的药物物质治疗或预防的另一种病理状况。
本发明还涉及一种抗生素失活酶，其用于在需要抗癌药剂的对象中阻止可能由抗生素引起的所述抗癌药剂效能的降低，所述抗生素被给药到所述对象是为了治疗或预防可以用所述抗生素治疗或预防的另一种病理状况。
在特定实施方式中，所述吸附剂或抗生素失活酶与引起生态失调的药物物质例如抗生素几乎同时给药到所述对象。“几乎同时”意味着所述吸附剂或抗生素失活酶在所述引起生态失调的药物物质特别是抗生素给药之前不久、给药的同时和/或给药之后不久，优选地在其给药之前不久给药。在特定实施方式中，所述吸附剂或抗生素失活酶在所述引起生态失调的药物物质特别是抗生素被给药之前或之后不到30分钟内给药，优选地在所述引起生态失调的药物物质特别是抗生素被给药之前或之后不到20分钟、不到19、18、17、16、15、14、13、12、11、10、9、8、7、6、5、4、3、2分钟或不到1分钟之内给药。在其他特定实施方式中，所述吸附剂或抗生素失活酶每天给药至少一次，特别是每天至少两次，更特别是每天三次或每天四次。在其他特定实施方式中，所述吸附剂或抗生素失活酶在使用所述引起生态失调的药物物质、特别是使用抗生素的整个治疗过程中给药。在这个实施方式的变化形式中，所述吸附剂或抗生素失活酶可以比所述引起生态失调的药物物质、特别是比抗生素给药更长的时间，以便确保消除任何残留的引起生态失调的药物物质，特别是任何残留的抗生素。例如，在中断所述引起生态失调的药物物质的给药后、特别是抗生素的给药后至少一天例如两天，仍然可以给药所述吸附剂或抗生素失活酶。
在特定实施方式中，本发明涉及一种吸附剂或抗生素失活酶，其与抗生素相组合、特别是几乎同时地用于需要抗癌药剂的对象。根据这个实施方式，所述吸附剂或抗生素失活酶阻止所述抗生素对所述对象的肠道微生物菌群可能具有的不良作用，并因此可以提高所述抗癌药剂的治疗效能。
因此，本发明还涉及一种包含吸附剂和引起生态失调的药物物质例如抗生素的药剂盒，或涉及一种包含抗生素失活酶和抗生素的药剂盒。所述药剂盒可用于治疗或预防可以使用所述引起生态失调的药物物质例如抗生素治疗或预防的病理状况。在所述药剂盒的特定实施方式中，所述引起生态失调的药物物质是抗生素。所述药剂盒还可以包含执行本发明的旨在阻止抗癌药剂的效能降低的方法的说明书。所述药剂盒的组分可以同时、分开地或顺序给药。正如上文中提供的，所述吸附剂或抗生素失活酶具体来说可以在所述引起生态失调的药剂例如抗生素给药之前、期间或之后给药，特别是在之前不久或之后不久、更特别是在之前不久给药。
实施例
实施例1：
为了评估在抗PD-1治疗期间使用抗生素的效果，将小鼠在第0天用癌细胞接种。小鼠从第-14天至第+25天通过皮下给药提供抗生素。在癌细胞接种后，在两周期间将小鼠通过腹膜内给药用抗PD-1治疗每周两次进行治疗。在所述实验期间，每两天记录肿瘤尺寸，并且也测量存活率。在第+25天，与未接受抗生素治疗的小鼠相比，在接受所述抗生素治疗的小鼠中观察到更大的肿瘤尺寸。
实施例2：
为了评估在抗PD-1治疗期间与抗生素相伴给药的吸附剂的效果，使用与实施例1中相同的流程，并从第-14天至第+28天通过口饲管每天两次提供吸附剂。在第+25天，与接受抗生素而没有吸附剂5的小鼠相比，在接受吸附剂5的小鼠中观察到更小的肿瘤尺寸。
实施例3：
为了评估在抗PD-1治疗期间与抗生素相伴给药的抗生素失活酶的效果，使用与实施例1中相同的流程，所述抗生素是β-内酰胺类抗生素。从第-14天至第+28天也通过口饲管每天两次提供β-内酰胺酶。在第+25天，与接受所述抗生素而没有β-内酰胺酶的小鼠相比，在接受所述β-内酰胺酶的小鼠中观察到更小的肿瘤尺寸。
实施例4：
为了评估在抗PD-L1治疗期间使用抗生素的效果，将小鼠在第0天用癌细胞接种。小鼠从第-14天至第+25天通过皮下给药提供抗生素。在癌细胞接种后，在两周期间将小鼠通过腹膜内给药用抗PD-L1治疗每周两次进行治疗。在所述实验期间，每两天记录肿瘤尺寸，并且也测量存活率。在第+25天，与未接受抗生素治疗的小鼠相比，在接受所述抗生素治疗的小鼠中观察到更大的肿瘤尺寸。
实施例5：
为了评估在抗PD-L1治疗期间与抗生素相伴给药的吸附剂的效果，使用与实施例3中相同的流程，并从第-14天至第+28天通过口饲管每天两次提供吸附剂。在第+25天，与接受抗生素而没有吸附剂的小鼠相比，在接受所述吸附剂的小鼠中观察到更小的肿瘤尺寸。
实施例6：
为了评估在抗PD-L1治疗期间与抗生素相伴给药的抗生素失活酶的效果，使用与实施例4中相同的流程，所述抗生素是β-内酰胺类抗生素。从第-14天至第+28天也通过口饲管每天两次提供β-内酰胺酶。在第+25天，与接受所述抗生素而没有β-内酰胺酶的小鼠相比，在接受所述β-内酰胺酶的小鼠中观察到更小的肿瘤尺寸。
实施例7：
为了评估在抗CTLA-4治疗期间使用抗生素的效果，将小鼠在第0天用癌细胞接种。小鼠从第-14天至第+25天通过皮下给药提供抗生素。在癌细胞接种后，在两周期间将小鼠通过腹膜内给药用抗CTLA-4治疗每周两次进行治疗。在所述实验期间，每两天记录肿瘤尺寸，并且也测量存活率。在第+25天，与未接受抗生素治疗的小鼠相比，在接受所述抗生素治疗的小鼠中观察到更大的肿瘤尺寸。
实施例8：
为了评估在抗CTLA-4治疗期间与抗生素相伴给药的吸附剂的效果，使用与实施例7中相同的流程，并从第-14天至第+28天通过口饲管每天两次提供吸附剂。在第+25天，与接受抗生素而没有吸附剂的小鼠相比，在接受所述吸附剂的小鼠中观察到更小的肿瘤尺寸。
实施例9：
为了评估在抗CTLA-4治疗期间与抗生素相伴给药的抗生素失活酶的效果，使用与实施例1中相同的流程，所述抗生素是β-内酰胺类抗生素。从第-14天至第+28天也通过口饲管每天两次提供β-内酰胺酶抗生素失活酶。在第+25天，与接受所述抗生素而没有β-内酰胺酶的小鼠相比，在接受所述β-内酰胺酶的小鼠中观察到更小的肿瘤尺寸。
实施例10：
为了评估在抗PD-1治疗期间与抗生素相伴给药的抗生素失活酶的效果，使用与实施例1中相同的流程，所述抗生素是大环内酯。从第-14天至第+28天也通过口饲管每天两次提供红霉素酯酶抗生素失活酶。在第+25天，与接受所述抗生素而没有红霉素酯酶的小鼠相比，在接受所述红霉素酯酶的小鼠中观察到更小的肿瘤尺寸。
实施例11：
为了评估在抗PD-L1治疗期间与抗生素相伴给药的抗生素失活酶的效果，使用与实施例1中相同的流程，所述抗生素是大环内酯。从第-14天至第+28天也通过口饲管每天两次提供红霉素酯酶抗生素失活酶。在第+25天，与接受所述抗生素而没有红霉素酯酶的小鼠相比，在接受所述红霉素酯酶的小鼠中观察到更小的肿瘤尺寸。
实施例12：
为了评估在抗CTLA-4治疗期间与抗生素相伴给药的抗生素失活酶的效果，使用与实施例1中相同的流程，所述抗生素是大环内酯。从第-14天至第+28天也通过口饲管每天两次提供红霉素酯酶抗生素失活酶。在第+25天，与接受所述抗生素而没有红霉素酯酶的小鼠相比，在接受所述红霉素酯酶的小鼠中观察到更小的肿瘤尺寸。
实施例13：
为了评估在抗PD-1治疗期间使用吸附剂的效果，将小鼠在第0天用癌细胞接种。在癌细胞接种后，在两周期间将小鼠通过腹膜内给药用抗PD-1治疗每周两次进行治疗。将所述小鼠分成两组，一组从第-14天至第+28天通过口饲管每天两次提供吸附剂，另一组不接受吸附剂。在所述实验期间，每两天记录肿瘤尺寸，并且也测量存活率。在第+25天，与接受吸附剂的小鼠相比，在未接受所述吸附剂的小鼠中观察到更大的肿瘤尺寸。
实施例14：
为了评估在抗PD-L1治疗期间使用吸附剂的效果，将小鼠在第0天用癌细胞接种。在癌细胞接种后，在两周期间将小鼠通过腹膜内给药用抗PD-L1治疗每周两次进行治疗。将所述小鼠分成两组，一组从第-14天至第+28天通过口饲管每天两次提供吸附剂，另一组不接受吸附剂。在所述实验期间，每两天记录肿瘤尺寸，并且也测量存活率。在第+25天，与接受吸附剂的小鼠相比，在未接受所述吸附剂的小鼠中观察到更大的肿瘤尺寸。
实施例15：
为了评估在抗CTLA-4治疗期间使用吸附剂的效果，将小鼠在第0天用癌细胞接种。在癌细胞接种后，在两周期间将小鼠通过腹膜内给药用抗CTLA-4治疗每周两次进行治疗。将所述小鼠分成两组，一组从第-14天至第+28天通过口饲管每天两次提供吸附剂，另一组不接受吸附剂。在所述实验期间，每两天记录肿瘤尺寸，并且也测量存活率。在第+25天，与接受吸附剂的小鼠相比，在未接受所述吸附剂的小鼠中观察到更大的肿瘤尺寸。
实施例16：
为了评估在抗PD1治疗期间使用吸附剂的能力，将60只小鼠在右前胁区域中用Hepa 1-6细胞(5x106)接种。肿瘤细胞接种的日期被命名为第0天。在肿瘤细胞接种后，每天检查动物的发病率和死亡率。在日常监测期间，检查所述动物的肿瘤生长和治疗对行为例如移动、食物和水的消耗、体重增加/减轻(体重在随机分组后每周测量两次)、眼/毛发失去光泽和任何其他异常的任何影响。为各个动物详细记录死亡率和观察到的临床体征。每周两次使用卡尺在两个维度上测量肿瘤体积。
将小鼠随机分成相等大小的3个组：
-组A：抗PD1抗体(3mg/kg)+抗生素安慰剂+吸附剂安慰剂
-组B：抗PD1抗体(3mg/kg)+克林霉素(25mg/kg)+吸附剂安慰剂
-组C：抗PD1抗体(3mg/kg)+克林霉素(25mg/kg)+吸附剂(1.5g/kg)
-组D：抗PD1抗体安慰剂(3mg/kg)+抗生素安慰剂+吸附剂安慰剂
所使用的抗PD1抗体是由Bioxcell生产的RMP1-14克隆。抗PD1抗体的同种型对照(抗PD1抗体安慰剂)是大鼠IgG2a。所述吸附剂是活性炭。克林霉素从GUANGZHOUBAIYUNSHAN TIANXIN PHARMACEUTICAL CO获得。所述抗PD1抗体每周两次腹膜内给药共3周。所述克林霉素从D-14到最后一次注射抗PD-1抗体后4天每天一次通过皮下途径给药。所述吸附剂从D-14到最后一次抗生素注射后3天每天两次通过口饲管给药。
我们在第29天测量了小鼠中的肿瘤尺寸，并将每个组的尺寸中位数报告在下表中：
Figure BDA0002921556420000391
从上表清楚地看出，与对照组D相比，在组A中抗PD1抗体能够降低肿瘤的生长。正如在本申请中提到的，在组B中抗生素的添加引起抗PD1抗体效能的丧失，在存在抗生素的情况下肿瘤生长如在对照组D中。令人吃惊的是，给药能够使抗生素残留物失活的吸附剂(参见组C)导致所述抗PD1抗体效能的恢复。因此，我们显示了吸附剂能够在给药抗生素的对象中提高抗癌药剂的效能。
序列表
<110> 达·沃尔泰拉公司（DA VOLTERRA）
<120> 提高抗癌药剂效能的方法
<130> B2811PC00
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225 230 235 240
<210> 11
<211> 235
<212> PRT
<213> 人工
<220>
<223> VIM-2 Q34R Q22H DCT236
<400> 11
Val Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn
225 230 235
<210> 12
<211> 235
<212> PRT
<213> 人工
<220>
<223> VIM-2 Q34R Q22N DCT236
<400> 12
Val Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn
225 230 235
<210> 13
<211> 235
<212> PRT
<213> 人工
<220>
<223> VIM-2 Q34R E130D DCT236
<400> 13
Val Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Gln Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn
225 230 235
<210> 14
<211> 235
<212> PRT
<213> 人工
<220>
<223> VIM-2 Q34R E130D Q22N DCT236
<400> 14
Val Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn
225 230 235
<210> 15
<211> 235
<212> PRT
<213> 人工
<220>
<223> VIM-2 Q34R E130D Q22H DCT236
<400> 15
Val Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn
225 230 235
<210> 16
<211> 240
<212> PRT
<213> 人工
<220>
<223> VIM-2 V1M Q34R Q22H
<400> 16
Met Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu
225 230 235 240
<210> 17
<211> 240
<212> PRT
<213> 人工
<220>
<223> VIM-2 V1M Q34R Q22N
<400> 17
Met Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu
225 230 235 240
<210> 18
<211> 240
<212> PRT
<213> 人工
<220>
<223> VIM-2 V1M Q34R E130D
<400> 18
Met Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Gln Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu
225 230 235 240
<210> 19
<211> 240
<212> PRT
<213> 人工
<220>
<223> VIM-2 V1M Q34R E130D Q22N
<400> 19
Met Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu
225 230 235 240
<210> 20
<211> 240
<212> PRT
<213> 人工
<220>
<223> VIM-2 V1M Q34R E130D Q22H
<400> 20
Met Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu
225 230 235 240
<210> 21
<211> 235
<212> PRT
<213> 人工
<220>
<223> VIM-2 V1M Q34R Q22H DCT236
<400> 21
Met Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn
225 230 235
<210> 22
<211> 235
<212> PRT
<213> 人工
<220>
<223> VIM-2 V1M Q34R Q22N DCT236
<400> 22
Met Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn
225 230 235
<210> 23
<211> 235
<212> PRT
<213> 人工
<220>
<223> VIM-2 V1M Q34R E130D DCT236
<400> 23
Met Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Gln Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn
225 230 235
<210> 24
<211> 235
<212> PRT
<213> 人工
<220>
<223> VIM-2 V1M Q34R E130D Q22N DCT236
<400> 24
Met Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn
225 230 235
<210> 25
<211> 235
<212> PRT
<213> 人工
<220>
<223> VIM-2 V1M Q34R E130D Q22H DCT236
<400> 25
Met Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn
225 230 235
<210> 26
<211> 240
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A
<400> 26
Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Gln Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Gln Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu
225 230 235 240
<210> 27
<211> 240
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A Q34R Q22H
<400> 27
Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu
225 230 235 240
<210> 28
<211> 240
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A Q34R Q22N
<400> 28
Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu
225 230 235 240
<210> 29
<211> 240
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A Q34R E130D
<400> 29
Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Gln Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu
225 230 235 240
<210> 30
<211> 240
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A Q34R E130D Q22N
<400> 30
Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu
225 230 235 240
<210> 31
<211> 240
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A Q34R E130D Q22H
<400> 31
Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu
225 230 235 240
<210> 32
<211> 235
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A Q34R Q22H DCT236
<400> 32
Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn
225 230 235
<210> 33
<211> 235
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A Q34R Q22N DCT236
<400> 33
Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn
225 230 235
<210> 34
<211> 235
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A Q34R E130D DCT236
<400> 34
Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Gln Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn
225 230 235
<210> 35
<211> 235
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A Q34R E130D Q22N DCT236
<400> 35
Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn
225 230 235
<210> 36
<211> 235
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A Q34R E130D Q22H DCT236
<400> 36
Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn
225 230 235
<210> 37
<211> 240
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A V1M Q34R Q22H
<400> 37
Met Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu
225 230 235 240
<210> 38
<211> 240
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A V1M Q34R Q22N
<400> 38
Met Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu
225 230 235 240
<210> 39
<211> 240
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A V1M Q34R E130D
<400> 39
Met Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Gln Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu
225 230 235 240
<210> 40
<211> 240
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A V1M Q34R E130D Q22N
<400> 40
Met Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu
225 230 235 240
<210> 41
<211> 240
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A V1M Q34R E130D Q22H
<400> 41
Met Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu
225 230 235 240
<210> 42
<211> 235
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A V1M Q34R Q22H DCT236
<400> 42
Met Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn
225 230 235
<210> 43
<211> 235
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A V1M Q34R Q22N DCT236
<400> 43
Met Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn
225 230 235
<210> 44
<211> 235
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A V1M Q34R E130D DCT236
<400> 44
Met Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Gln Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn
225 230 235
<210> 45
<211> 235
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A V1M Q34R E130D Q22N DCT236
<400> 45
Met Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn
225 230 235
<210> 46
<211> 235
<212> PRT
<213> 人工
<220>
<223> VIM-2 V10A V1M Q34R E130D Q22H DCT236
<400> 46
Met Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly
1 5 10 15
Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala
20 25 30
Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val
35 40 45
Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys
50 55 60
Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro
65 70 75 80
Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly
85 90 95
Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser
100 105 110
Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser
115 120 125
Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val
130 135 140
Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val
145 150 155 160
Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu
165 170 175
Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu
180 185 190
Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln
195 200 205
Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys
210 215 220
His Thr Thr Asn Val Val Lys Ala His Thr Asn
225 230 235
<210> 47
<211> 263
<212> PRT
<213> 人工
<220>
<223> P1A
<400> 47
Glu Met Lys Asp Asp Phe Ala Lys Leu Glu Glu Gln Phe Asp Ala Lys
1 5 10 15
Leu Gly Ile Phe Ala Leu Asp Thr Gly Thr Asn Arg Thr Val Ala Tyr
20 25 30
Arg Pro Asp Glu Arg Phe Ala Phe Ala Ser Thr Ile Lys Ala Leu Thr
35 40 45
Val Gly Val Leu Leu Gln Gln Lys Ser Ile Glu Asp Leu Asn Gln Arg
50 55 60
Ile Thr Tyr Thr Arg Asp Asp Leu Val Asn Tyr Asn Pro Ile Thr Glu
65 70 75 80
Lys His Val Asp Thr Gly Met Thr Leu Lys Glu Leu Ala Asp Ala Ser
85 90 95
Leu Arg Tyr Ser Asp Asn Ala Ala Gln Asn Leu Ile Leu Lys Gln Ile
100 105 110
Gly Gly Pro Glu Ser Leu Lys Lys Glu Leu Arg Lys Ile Gly Asp Glu
115 120 125
Val Thr Asn Pro Glu Arg Phe Glu Pro Glu Leu Asn Glu Val Asn Pro
130 135 140
Gly Glu Thr Gln Asp Thr Ser Thr Ala Arg Ala Leu Val Thr Ser Leu
145 150 155 160
Arg Ala Phe Ala Leu Glu Asp Lys Leu Pro Ser Glu Lys Arg Glu Leu
165 170 175
Leu Ile Asp Trp Met Lys Arg Asn Thr Thr Gly Asp Ala Leu Ile Arg
180 185 190
Ala Gly Val Pro Asp Gly Trp Glu Val Ala Asp Lys Thr Gly Ala Ala
195 200 205
Ser Tyr Gly Thr Arg Asn Asp Ile Ala Ile Ile Trp Pro Pro Lys Gly
210 215 220
Asp Pro Val Val Leu Ala Val Leu Ser Ser Arg Asp Lys Lys Asp Ala
225 230 235 240
Lys Tyr Asp Asp Lys Leu Ile Ala Glu Ala Thr Lys Val Val Met Lys
245 250 255
Ala Leu Asn Met Asn Gly Lys
260
<210> 48
<211> 215
<212> PRT
<213> 人工
<220>
<223> P2A
<400> 48
Glu Thr Gly Thr Ile Ser Ile Ser Gln Leu Asn Lys Asn Val Trp Val
1 5 10 15
His Thr Glu Leu Gly Tyr Phe Asn Gly Glu Ala Val Pro Ser Asn Gly
20 25 30
Leu Val Leu Asn Thr Ser Lys Gly Leu Val Leu Val Asp Ser Ser Trp
35 40 45
Asp Asn Lys Leu Thr Lys Glu Leu Ile Glu Met Val Glu Lys Lys Phe
50 55 60
Gln Lys Arg Val Thr Asp Val Ile Ile Thr His Ala His Ala Asp Arg
65 70 75 80
Ile Gly Gly Ile Thr Ala Leu Lys Glu Arg Gly Ile Lys Ala His Ser
85 90 95
Thr Ala Leu Thr Ala Glu Leu Ala Lys Asn Ser Gly Tyr Glu Glu Pro
100 105 110
Leu Gly Asp Leu Gln Thr Ile Thr Ser Leu Lys Phe Gly Asn Thr Lys
115 120 125
Val Glu Thr Phe Tyr Pro Gly Lys Gly His Thr Glu Asp Asn Ile Val
130 135 140
Val Trp Leu Pro Gln Tyr Gln Ile Leu Ala Gly Gly Cys Leu Val Lys
145 150 155 160
Ser Ala Glu Ala Lys Asp Leu Gly Asn Val Ala Asp Ala Tyr Val Asn
165 170 175
Glu Trp Ser Thr Ser Ile Glu Asn Val Leu Lys Arg Tyr Gly Asn Ile
180 185 190
Asn Ser Trp Pro Gly His Gly Glu Val Gly Asp Lys Gly Leu Leu Leu
195 200 205
His Thr Leu Asp Leu Leu Lys
210 215
<210> 49
<211> 258
<212> PRT
<213> 人工
<220>
<223> P3A变体
<220>
<221> MISC_FEATURE
<222> (243)..(243)
<223> Xaa可以是任何天然存在的氨基酸
<400> 49
Met Lys Asp Asp Phe Ala Lys Leu Glu Glu Gln Phe Asp Ala Lys Leu
1 5 10 15
Gly Ile Phe Ala Leu Asp Thr Gly Thr Asn Arg Thr Val Ala Tyr Arg
20 25 30
Pro Asp Glu Arg Phe Ala Phe Ala Ser Thr Ile Lys Ala Leu Thr Val
35 40 45
Gly Val Leu Leu Gln Gln Lys Ser Ile Glu Asp Leu Asn Gln Arg Ile
50 55 60
Thr Tyr Thr Arg Asp Asp Leu Val Asn Tyr Asn Pro Ile Thr Glu Lys
65 70 75 80
His Val Asp Thr Gly Met Thr Leu Lys Glu Leu Ala Asp Ala Ser Leu
85 90 95
Arg Tyr Ser Asp Asn Ala Ala Gln Asn Leu Ile Leu Lys Gln Ile Gly
100 105 110
Gly Pro Glu Ser Leu Lys Lys Glu Leu Arg Lys Ile Gly Asp Glu Val
115 120 125
Thr Asn Pro Glu Arg Phe Glu Pro Glu Leu Asn Glu Val Asn Pro Gly
130 135 140
Glu Thr Gln Asp Thr Ser Thr Ala Arg Ala Leu Val Thr Ser Leu Arg
145 150 155 160
Ala Phe Ala Leu Glu Asp Lys Leu Pro Ser Glu Lys Arg Glu Leu Leu
165 170 175
Ile Asp Trp Met Lys Arg Asn Thr Thr Gly Asp Ala Leu Ile Arg Ala
180 185 190
Gly Val Pro Asp Gly Trp Glu Val Ala Asp Lys Thr Gly Ala Ala Ser
195 200 205
Tyr Gly Thr Arg Asn Asp Ile Ala Ile Ile Trp Pro Pro Lys Gly Asp
210 215 220
Pro Val Val Leu Ala Val Leu Ser Ser Arg Asp Lys Lys Asp Ala Lys
225 230 235 240
Tyr Asp Xaa Lys Leu Ile Ala Glu Ala Thr Lys Val Val Met Lys Ala
245 250 255
Leu Asn
<210> 50
<211> 299
<212> PRT
<213> 人工
<220>
<223> P3A变体
<220>
<221> misc_feature
<222> (280)..(280)
<223> Xaa可以是任何天然存在的氨基酸
<220>
<221> MISC_FEATURE
<222> (290)..(290)
<223> Xaa可以是任何天然存在的氨基酸
<400> 50
Met Ile Gln Lys Arg Lys Arg Thr Val Ser Phe Arg Leu Val Leu Met
1 5 10 15
Cys Thr Leu Leu Phe Val Ser Leu Pro Ile Thr Lys Thr Ser Ala Gln
20 25 30
Ala Ser Lys Thr Glu Met Lys Asp Asp Phe Ala Lys Leu Glu Glu Gln
35 40 45
Phe Asp Ala Lys Leu Gly Ile Phe Ala Leu Asp Thr Gly Thr Asn Arg
50 55 60
Thr Val Ala Tyr Arg Pro Asp Glu Arg Phe Ala Phe Ala Ser Thr Ile
65 70 75 80
Lys Ala Leu Thr Val Gly Val Leu Leu Gln Gln Lys Ser Ile Glu Asp
85 90 95
Leu Asn Gln Arg Ile Thr Tyr Thr Arg Asp Asp Leu Val Asn Tyr Asn
100 105 110
Pro Ile Thr Glu Lys His Val Asp Thr Gly Met Thr Leu Lys Glu Leu
115 120 125
Ala Asp Ala Ser Leu Arg Tyr Ser Asp Asn Ala Ala Gln Asn Leu Ile
130 135 140
Leu Lys Gln Ile Gly Gly Pro Glu Ser Leu Lys Lys Glu Leu Arg Lys
145 150 155 160
Ile Gly Asp Glu Val Thr Asn Pro Glu Arg Phe Glu Pro Glu Leu Asn
165 170 175
Glu Val Asn Pro Gly Glu Thr Gln Asp Thr Ser Thr Ala Arg Ala Leu
180 185 190
Val Thr Ser Leu Arg Ala Phe Ala Leu Glu Asp Lys Leu Pro Ser Glu
195 200 205
Lys Arg Glu Leu Leu Ile Asp Trp Met Lys Arg Asn Thr Thr Gly Asp
210 215 220
Ala Leu Ile Arg Ala Gly Val Pro Asp Gly Trp Glu Val Ala Asp Lys
225 230 235 240
Thr Gly Ala Ala Ser Tyr Gly Thr Arg Asn Asp Ile Ala Ile Ile Trp
245 250 255
Pro Pro Lys Gly Asp Pro Val Val Leu Ala Val Leu Ser Ser Arg Asp
260 265 270
Lys Lys Asp Ala Lys Tyr Asp Xaa Lys Leu Ile Ala Glu Ala Thr Lys
275 280 285
Val Val Met Lys Ala Leu Asn Met Asn Gly Lys
290 295
<210> 51
<211> 293
<212> PRT
<213> 人工
<220>
<223> KPC-1/2
<400> 51
Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro
1 5 10 15
Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro
20 25 30
Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala
35 40 45
Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg
50 55 60
Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu
65 70 75 80
Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr
85 90 95
Gly Lys Asn Ala Leu Val Pro Trp Ser Pro Ile Ser Glu Lys Tyr Leu
100 105 110
Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr
115 120 125
Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro
130 135 140
Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg
145 150 155 160
Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Ser Asp Ala
165 170 175
Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu
180 185 190
Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp
195 200 205
Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val
210 215 220
Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Val Tyr
225 230 235 240
Gly Thr Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro
245 250 255
Ile Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys His
260 265 270
Ser Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu
275 280 285
Gly Val Asn Gly Gln
290
<210> 52
<211> 293
<212> PRT
<213> 人工
<220>
<223> KPC-3
<400> 52
Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro
1 5 10 15
Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro
20 25 30
Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala
35 40 45
Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg
50 55 60
Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu
65 70 75 80
Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr
85 90 95
Gly Lys Asn Ala Leu Val Pro Trp Ser Pro Ile Ser Glu Lys Tyr Leu
100 105 110
Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr
115 120 125
Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro
130 135 140
Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg
145 150 155 160
Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala
165 170 175
Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu
180 185 190
Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp
195 200 205
Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val
210 215 220
Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Val Tyr
225 230 235 240
Gly Thr Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro
245 250 255
Ile Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys Tyr
260 265 270
Ser Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu
275 280 285
Gly Val Asn Gly Gln
290
<210> 53
<211> 292
<212> PRT
<213> 人工
<220>
<223> KPC-4
<400> 53
Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro
1 5 10 15
Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro
20 25 30
Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala
35 40 45
Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg
50 55 60
Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu
65 70 75 80
Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr
85 90 95
Gly Lys Asn Ala Leu Val Arg Trp Ser Pro Ile Ser Glu Lys Tyr Leu
100 105 110
Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr
115 120 125
Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro
130 135 140
Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg
145 150 155 160
Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala
165 170 175
Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu
180 185 190
Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp
195 200 205
Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val
210 215 220
Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Gly Tyr
225 230 235 240
Gly Thr Ala Asn Asp Tyr Ala Trp Trp Pro Thr Gly Arg Ala Pro Ile
245 250 255
Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys His Ser
260 265 270
Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu Gly
275 280 285
Val Asn Gly Gln
290
<210> 54
<211> 293
<212> PRT
<213> 人工
<220>
<223> KPC-5
<400> 54
Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro
1 5 10 15
Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro
20 25 30
Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala
35 40 45
Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg
50 55 60
Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu
65 70 75 80
Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr
85 90 95
Gly Lys Asn Ala Leu Val Arg Trp Ser Pro Ile Ser Glu Lys Tyr Leu
100 105 110
Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr
115 120 125
Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro
130 135 140
Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg
145 150 155 160
Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala
165 170 175
Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu
180 185 190
Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp
195 200 205
Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val
210 215 220
Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Val Tyr
225 230 235 240
Gly Thr Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro
245 250 255
Ile Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys His
260 265 270
Ser Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu
275 280 285
Gly Val Asn Gly Gln
290
<210> 55
<211> 292
<212> PRT
<213> 人工
<220>
<223> KPC-6
<400> 55
Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro
1 5 10 15
Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro
20 25 30
Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala
35 40 45
Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg
50 55 60
Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu
65 70 75 80
Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr
85 90 95
Gly Lys Asn Ala Leu Val Pro Trp Ser Pro Ile Ser Glu Lys Tyr Leu
100 105 110
Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr
115 120 125
Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro
130 135 140
Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg
145 150 155 160
Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala
165 170 175
Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu
180 185 190
Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp
195 200 205
Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val
210 215 220
Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Gly Tyr
225 230 235 240
Gly Thr Ala Asn Asp Tyr Ala Trp Trp Pro Thr Gly Arg Ala Pro Ile
245 250 255
Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys His Ser
260 265 270
Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu Gly
275 280 285
Val Asn Gly Gln
290
<210> 56
<211> 293
<212> PRT
<213> 人工
<220>
<223> KPC-7
<400> 56
Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro
1 5 10 15
Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro
20 25 30
Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala
35 40 45
Ile Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg
50 55 60
Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu
65 70 75 80
Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr
85 90 95
Gly Lys Asn Ala Leu Val Pro Trp Ser Pro Ile Ser Glu Lys Tyr Leu
100 105 110
Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr
115 120 125
Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro
130 135 140
Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg
145 150 155 160
Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala
165 170 175
Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu
180 185 190
Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp
195 200 205
Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Leu Arg Ala Ala Val
210 215 220
Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Val Tyr
225 230 235 240
Gly Thr Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro
245 250 255
Ile Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys Tyr
260 265 270
Ser Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu
275 280 285
Gly Val Asn Gly Gln
290
<210> 57
<211> 292
<212> PRT
<213> 人工
<220>
<223> KPC-8
<400> 57
Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro
1 5 10 15
Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro
20 25 30
Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala
35 40 45
Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg
50 55 60
Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu
65 70 75 80
Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr
85 90 95
Gly Lys Asn Ala Leu Val Pro Trp Ser Pro Ile Ser Glu Lys Tyr Leu
100 105 110
Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr
115 120 125
Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro
130 135 140
Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg
145 150 155 160
Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala
165 170 175
Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu
180 185 190
Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp
195 200 205
Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val
210 215 220
Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Gly Tyr
225 230 235 240
Gly Thr Ala Asn Asp Tyr Ala Trp Trp Pro Thr Gly Arg Ala Pro Ile
245 250 255
Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys Tyr Ser
260 265 270
Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu Gly
275 280 285
Val Asn Gly Gln
290
<210> 58
<211> 293
<212> PRT
<213> 人工
<220>
<223> KPC-9
<400> 58
Met Ser Lys Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro
1 5 10 15
Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro
20 25 30
Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala
35 40 45
Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg
50 55 60
Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu
65 70 75 80
Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr
85 90 95
Gly Lys Asn Ala Leu Val Pro Trp Ser Pro Ile Ser Glu Lys Tyr Leu
100 105 110
Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr
115 120 125
Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro
130 135 140
Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg
145 150 155 160
Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala
165 170 175
Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu
180 185 190
Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp
195 200 205
Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val
210 215 220
Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Ala Tyr
225 230 235 240
Gly Thr Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro
245 250 255
Ile Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys Tyr
260 265 270
Ser Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu
275 280 285
Gly Val Asn Gly Gln
290
<210> 59
<211> 293
<212> PRT
<213> 人工
<220>
<223> KPC-10
<400> 59
Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro
1 5 10 15
Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro
20 25 30
Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala
35 40 45
Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg
50 55 60
Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu
65 70 75 80
Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr
85 90 95
Gly Lys Asn Ala Leu Val Arg Trp Ser Pro Ile Ser Glu Lys Tyr Leu
100 105 110
Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr
115 120 125
Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro
130 135 140
Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg
145 150 155 160
Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala
165 170 175
Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu
180 185 190
Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp
195 200 205
Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val
210 215 220
Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Val Tyr
225 230 235 240
Gly Thr Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro
245 250 255
Ile Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys Tyr
260 265 270
Ser Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu
275 280 285
Gly Val Asn Gly Gln
290
<210> 60
<211> 293
<212> PRT
<213> 人工
<220>
<223> KPC-11
<400> 60
Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro
1 5 10 15
Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro
20 25 30
Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala
35 40 45
Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg
50 55 60
Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu
65 70 75 80
Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr
85 90 95
Gly Lys Asn Ala Leu Val Leu Trp Ser Pro Ile Ser Glu Lys Tyr Leu
100 105 110
Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr
115 120 125
Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro
130 135 140
Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg
145 150 155 160
Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala
165 170 175
Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu
180 185 190
Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp
195 200 205
Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val
210 215 220
Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Val Tyr
225 230 235 240
Gly Thr Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro
245 250 255
Ile Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys His
260 265 270
Ser Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu
275 280 285
Gly Val Asn Gly Gln
290
<210> 61
<211> 293
<212> PRT
<213> 人工
<220>
<223> KPC-12
<400> 61
Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro
1 5 10 15
Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro
20 25 30
Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala
35 40 45
Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg
50 55 60
Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu
65 70 75 80
Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr
85 90 95
Gly Lys Asn Ala Leu Val Pro Trp Ser Pro Ile Ser Glu Lys Tyr Leu
100 105 110
Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr
115 120 125
Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro
130 135 140
Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg
145 150 155 160
Leu Asp Arg Trp Glu Leu Glu Met Asn Ser Ala Ile Pro Gly Asp Ala
165 170 175
Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu
180 185 190
Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp
195 200 205
Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val
210 215 220
Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Val Tyr
225 230 235 240
Gly Thr Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro
245 250 255
Ile Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys His
260 265 270
Ser Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu
275 280 285
Gly Val Asn Gly Gln
290
<210> 62
<211> 293
<212> PRT
<213> 人工
<220>
<223> KPC-13
<400> 62
Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro
1 5 10 15
Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro
20 25 30
Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala
35 40 45
Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg
50 55 60
Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu
65 70 75 80
Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Gly Thr Pro Ile Arg Tyr
85 90 95
Gly Lys Asn Ala Leu Val Pro Trp Ser Pro Ile Ser Glu Lys Tyr Leu
100 105 110
Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr
115 120 125
Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro
130 135 140
Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg
145 150 155 160
Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala
165 170 175
Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu
180 185 190
Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp
195 200 205
Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val
210 215 220
Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Val Tyr
225 230 235 240
Gly Thr Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro
245 250 255
Ile Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys Tyr
260 265 270
Ser Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu
275 280 285
Gly Val Asn Gly Gln
290
<210> 63
<211> 291
<212> PRT
<213> 人工
<220>
<223> KPC-14
<400> 63
Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro
1 5 10 15
Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro
20 25 30
Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala
35 40 45
Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg
50 55 60
Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu
65 70 75 80
Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr
85 90 95
Gly Lys Asn Ala Leu Val Pro Trp Ser Pro Ile Ser Glu Lys Tyr Leu
100 105 110
Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr
115 120 125
Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro
130 135 140
Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg
145 150 155 160
Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala
165 170 175
Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu
180 185 190
Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp
195 200 205
Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val
210 215 220
Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Val Tyr
225 230 235 240
Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro Ile Val
245 250 255
Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys His Ser Glu
260 265 270
Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu Gly Val
275 280 285
Asn Gly Gln
290
<210> 64
<211> 292
<212> PRT
<213> 人工
<220>
<223> KPC-15
<400> 64
Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro
1 5 10 15
Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro
20 25 30
Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala
35 40 45
Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg
50 55 60
Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu
65 70 75 80
Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr
85 90 95
Gly Lys Asn Ala Leu Val Arg Trp Ser Pro Ile Ser Glu Lys Tyr Leu
100 105 110
Thr Thr Gly Met Thr Val Leu Glu Leu Ser Ala Ala Ala Val Gln Tyr
115 120 125
Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro
130 135 140
Ala Lys Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg
145 150 155 160
Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala
165 170 175
Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu
180 185 190
Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp
195 200 205
Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val
210 215 220
Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Gly Tyr
225 230 235 240
Gly Thr Ala Asn Asp Tyr Ala Trp Trp Pro Thr Gly Arg Ala Pro Ile
245 250 255
Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys Tyr Ser
260 265 270
Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu Gly
275 280 285
Val Asn Gly Gln
290
<210> 65
<211> 293
<212> PRT
<213> 人工
<220>
<223> KPC-17
<400> 65
Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro
1 5 10 15
Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro
20 25 30
Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala
35 40 45
Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg
50 55 60
Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu
65 70 75 80
Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr
85 90 95
Gly Lys Asn Ala Leu Val Pro Trp Ser Pro Ile Ser Glu Lys Tyr Leu
100 105 110
Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr
115 120 125
Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro
130 135 140
Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg
145 150 155 160
Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala
165 170 175
Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu
180 185 190
Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Leu Val Asp
195 200 205
Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val
210 215 220
Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Val Tyr
225 230 235 240
Gly Thr Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro
245 250 255
Ile Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys His
260 265 270
Ser Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu
275 280 285
Gly Val Asn Gly Gln
290
<210> 66
<211> 269
<212> PRT
<213> 人工
<220>
<223> NDM-1
<400> 66
Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala
1 5 10 15
Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg
20 25 30
Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp
35 40 45
Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr
50 55 60
Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val
65 70 75 80
Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asp Asp
85 90 95
Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro
100 105 110
Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met
115 120 125
Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser
130 135 140
Asn Gln Leu Ala Pro Gln Glu Gly Met Val Ala Ala Gln His Ser Leu
145 150 155 160
Thr Phe Ala Ala Asn Gly Val Trp Glu Pro Ala Thr Ala Pro Asn Phe
165 170 175
Gly Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn
180 185 190
Ile Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu
195 200 205
Ile Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala Asp
210 215 220
Thr Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro
225 230 235 240
Lys Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala
245 250 255
Ala Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg
260 265
<210> 67
<211> 269
<212> PRT
<213> 人工
<220>
<223> NDM-2
<400> 67
Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala
1 5 10 15
Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Ala Gly Glu Ile Arg
20 25 30
Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp
35 40 45
Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr
50 55 60
Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val
65 70 75 80
Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asp Asp
85 90 95
Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro
100 105 110
Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met
115 120 125
Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser
130 135 140
Asn Gln Leu Ala Pro Gln Glu Gly Met Val Ala Ala Gln His Ser Leu
145 150 155 160
Thr Phe Ala Ala Asn Gly Val Trp Glu Pro Ala Thr Ala Pro Asn Phe
165 170 175
Gly Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn
180 185 190
Ile Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu
195 200 205
Ile Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala Asp
210 215 220
Thr Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro
225 230 235 240
Lys Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala
245 250 255
Ala Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg
260 265
<210> 68
<211> 269
<212> PRT
<213> 人工
<220>
<223> NDM-3
<400> 68
Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala
1 5 10 15
Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg
20 25 30
Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp
35 40 45
Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr
50 55 60
Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val
65 70 75 80
Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asn Asp
85 90 95
Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro
100 105 110
Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met
115 120 125
Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser
130 135 140
Asn Gln Leu Ala Pro Gln Glu Gly Met Val Ala Ala Gln His Ser Leu
145 150 155 160
Thr Phe Ala Ala Asn Gly Val Trp Glu Pro Ala Thr Ala Pro Asn Phe
165 170 175
Gly Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn
180 185 190
Ile Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu
195 200 205
Ile Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala Asp
210 215 220
Thr Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro
225 230 235 240
Lys Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala
245 250 255
Ala Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg
260 265
<210> 69
<211> 268
<212> PRT
<213> 人工
<220>
<223> NDM-4
<400> 69
Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala
1 5 10 15
Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg
20 25 30
Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp
35 40 45
Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr
50 55 60
Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val
65 70 75 80
Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asp Asp
85 90 95
Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro
100 105 110
Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met
115 120 125
Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser
130 135 140
Asn Gln Leu Ala Pro Gln Glu Gly Leu Val Ala Ala Gln His Ser Leu
145 150 155 160
Thr Phe Ala Ala Asn Gly Trp Glu Pro Ala Thr Ala Pro Asn Phe Gly
165 170 175
Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn Ile
180 185 190
Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu Ile
195 200 205
Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala Asp Thr
210 215 220
Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro Lys
225 230 235 240
Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala Ala
245 250 255
Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg
260 265
<210> 70
<211> 270
<212> PRT
<213> 人工
<220>
<223> NDM-5
<400> 70
Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala
1 5 10 15
Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg
20 25 30
Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp
35 40 45
Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr
50 55 60
Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val
65 70 75 80
Arg Asp Gly Gly Arg Val Leu Leu Val Asp Thr Ala Trp Thr Asp Asp
85 90 95
Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro
100 105 110
Val Ala Leu Ala Val Val Thr His Ala His Gln Asp Lys Met Gly Gly
115 120 125
Met Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu
130 135 140
Ser Asn Gln Leu Ala Pro Gln Glu Gly Leu Val Ala Ala Gln His Ser
145 150 155 160
Leu Thr Phe Ala Ala Asn Gly Val Trp Glu Pro Ala Thr Ala Pro Asn
165 170 175
Phe Gly Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp
180 185 190
Asn Ile Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys
195 200 205
Leu Ile Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala
210 215 220
Asp Thr Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe
225 230 235 240
Pro Lys Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg
245 250 255
Ala Ala Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg
260 265 270
<210> 71
<211> 269
<212> PRT
<213> 人工
<220>
<223> NDM-6
<400> 71
Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala
1 5 10 15
Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg
20 25 30
Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp
35 40 45
Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr
50 55 60
Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val
65 70 75 80
Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asp Asp
85 90 95
Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro
100 105 110
Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met
115 120 125
Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser
130 135 140
Asn Gln Leu Ala Pro Gln Glu Gly Met Val Ala Ala Gln His Ser Leu
145 150 155 160
Thr Phe Ala Ala Asn Gly Val Trp Glu Pro Ala Thr Ala Pro Asn Phe
165 170 175
Gly Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn
180 185 190
Ile Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu
195 200 205
Ile Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala Asp
210 215 220
Thr Glu His Tyr Ala Ala Ser Val Arg Ala Phe Gly Ala Ala Phe Pro
225 230 235 240
Lys Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala
245 250 255
Ala Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg
260 265
<210> 72
<211> 268
<212> PRT
<213> 人工
<220>
<223> NDM-7
<400> 72
Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala
1 5 10 15
Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg
20 25 30
Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp
35 40 45
Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr
50 55 60
Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val
65 70 75 80
Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asp Asp
85 90 95
Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro
100 105 110
Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met
115 120 125
Asn Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser
130 135 140
Asn Gln Leu Ala Pro Gln Glu Gly Leu Val Ala Ala Gln His Ser Leu
145 150 155 160
Thr Phe Ala Ala Asn Gly Trp Glu Pro Ala Thr Ala Pro Asn Phe Gly
165 170 175
Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn Ile
180 185 190
Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu Ile
195 200 205
Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala Asp Thr
210 215 220
Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro Lys
225 230 235 240
Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala Ala
245 250 255
Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg
260 265
<210> 73
<211> 269
<212> PRT
<213> 人工
<220>
<223> NDM-8
<400> 73
Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala
1 5 10 15
Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg
20 25 30
Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp
35 40 45
Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr
50 55 60
Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val
65 70 75 80
Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asp Asp
85 90 95
Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro
100 105 110
Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met
115 120 125
Gly Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser
130 135 140
Asn Gln Leu Ala Pro Gln Glu Gly Leu Val Ala Ala Gln His Ser Leu
145 150 155 160
Thr Phe Ala Ala Asn Gly Val Trp Glu Pro Ala Thr Ala Pro Asn Phe
165 170 175
Gly Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn
180 185 190
Ile Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu
195 200 205
Ile Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala Asp
210 215 220
Thr Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro
225 230 235 240
Lys Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala
245 250 255
Ala Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg
260 265
<210> 74
<211> 268
<212> PRT
<213> 人工
<220>
<223> NDM-9
<400> 74
Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala
1 5 10 15
Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg
20 25 30
Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp
35 40 45
Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr
50 55 60
Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val
65 70 75 80
Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asp Asp
85 90 95
Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro
100 105 110
Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met
115 120 125
Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser
130 135 140
Asn Gln Leu Ala Pro Gln Lys Gly Met Val Ala Ala Gln His Ser Leu
145 150 155 160
Thr Phe Ala Ala Asn Gly Trp Glu Pro Ala Thr Ala Pro Asn Phe Gly
165 170 175
Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn Ile
180 185 190
Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu Ile
195 200 205
Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala Asp Thr
210 215 220
Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro Lys
225 230 235 240
Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala Ala
245 250 255
Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg
260 265
<210> 75
<211> 269
<212> PRT
<213> 人工
<220>
<223> NDM-10
<400> 75
Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala
1 5 10 15
Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Ser
20 25 30
Pro Thr Ile Asp Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp
35 40 45
Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr
50 55 60
Leu Asp Met Pro Ser Phe Gly Ala Val Thr Ser Asn Gly Leu Ile Val
65 70 75 80
Arg Asp Gly Gly Arg Val Leu Trp Asp Thr Ala Trp Thr Asp Asp Gln
85 90 95
Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro Val
100 105 110
Ala Leu Ala Val Val Thr His Ala His Gln Asp Lys Met Gly Gly Met
115 120 125
Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser
130 135 140
Asn Gln Leu Ala Pro Gln Glu Gly Met Val Ala Ala Gln His Ser Leu
145 150 155 160
Thr Phe Ala Ala Asn Gly Trp Val Glu Pro Ala Thr Ala Pro Asn Phe
165 170 175
Gly Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn
180 185 190
Ile Thr Val Gly Ile Asp Arg Thr Asp Ile Ala Phe Gly Gly Cys Leu
195 200 205
Ile Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala Asp
210 215 220
Thr Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro
225 230 235 240
Lys Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala
245 250 255
Ala Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg
260 265
<210> 76
<211> 268
<212> PRT
<213> 人工
<220>
<223> NDM-11
<400> 76
Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala
1 5 10 15
Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg
20 25 30
Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp
35 40 45
Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr
50 55 60
Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val
65 70 75 80
Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asp Asp
85 90 95
Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro
100 105 110
Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met
115 120 125
Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser
130 135 140
Asn Gln Leu Ala Pro Gln Glu Gly Leu Val Ala Ala Gln His Ser Leu
145 150 155 160
Thr Phe Ala Ala Asn Gly Trp Glu Pro Ala Thr Ala Pro Asn Phe Gly
165 170 175
Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn Ile
180 185 190
Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu Ile
195 200 205
Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Asp Asp Ala Asp Thr
210 215 220
Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro Lys
225 230 235 240
Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala Ala
245 250 255
Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg
260 265
<210> 77
<211> 268
<212> PRT
<213> 人工
<220>
<223> NDM-12
<400> 77
Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala
1 5 10 15
Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg
20 25 30
Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp
35 40 45
Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr
50 55 60
Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val
65 70 75 80
Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asp Asp
85 90 95
Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro
100 105 110
Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met
115 120 125
Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser
130 135 140
Asn Gln Leu Ala Pro Gln Glu Gly Leu Val Ala Ala Gln His Ser Leu
145 150 155 160
Thr Phe Ala Ala Asn Gly Trp Glu Pro Ala Thr Ala Pro Asn Phe Gly
165 170 175
Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn Ile
180 185 190
Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu Ile
195 200 205
Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Asp Asp Ala Asp Thr
210 215 220
Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro Lys
225 230 235 240
Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala Ala
245 250 255
Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg
260 265
<210> 78
<211> 268
<212> PRT
<213> 人工
<220>
<223> NDM-13
<400> 78
Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala
1 5 10 15
Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg
20 25 30
Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp
35 40 45
Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr
50 55 60
Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val
65 70 75 80
Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asn Asp
85 90 95
Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro
100 105 110
Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met
115 120 125
Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser
130 135 140
Asn Gln Leu Ala Pro Gln Glu Gly Leu Val Ala Ala Gln His Ser Leu
145 150 155 160
Thr Phe Ala Ala Asn Gly Trp Glu Pro Ala Thr Ala Pro Asn Phe Gly
165 170 175
Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn Ile
180 185 190
Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu Ile
195 200 205
Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala Asp Thr
210 215 220
Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro Lys
225 230 235 240
Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala Ala
245 250 255
Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg
260 265
<210> 79
<211> 260
<212> PRT
<213> 人工
<220>
<223> P4A
<400> 79
Glu Met Lys Asp Asp Phe Ala Lys Leu Glu Glu Gln Phe Asp Ala Lys
1 5 10 15
Leu Gly Ile Phe Ala Leu Asp Thr Gly Thr Asn Arg Thr Val Ala Tyr
20 25 30
Arg Pro Asp Glu Arg Phe Ala Phe Ala Ser Thr Ile Lys Ala Leu Thr
35 40 45
Val Gly Val Leu Leu Gln Gln Lys Ser Ile Glu Asp Leu Asn Gln Arg
50 55 60
Ile Thr Thr Arg Asp Asp Leu Val Asn Tyr Asn Pro Ile Thr Glu Lys
65 70 75 80
His Val Asp Thr Gly Met Thr Leu Lys Glu Leu Ala Asp Ala Ser Leu
85 90 95
Arg Tyr Ser Asp Asn Ala Ala Gln Asn Leu Ile Leu Lys Gln Ile Gly
100 105 110
Gly Pro Glu Ser Leu Lys Lys Glu Leu Arg Lys Ile Gly Asp Glu Val
115 120 125
Thr Asn Pro Glu Arg Phe Glu Pro Glu Leu Asn Glu Val Asn Pro Gly
130 135 140
Glu Thr Gln Asp Thr Ser Thr Ala Arg Ala Leu Val Thr Ser Leu Arg
145 150 155 160
Ala Phe Ala Leu Glu Asp Lys Leu Pro Ser Glu Lys Arg Glu Leu Leu
165 170 175
Ile Asp Trp Met Lys Arg Asn Thr Thr Gly Asp Ala Leu Ile Arg Ala
180 185 190
Gly Val Pro Asp Gly Trp Glu Val Gly Asp Lys Thr Gly Ser Gly Asp
195 200 205
Tyr Gly Thr Arg Asn Asp Ile Ala Ile Ile Trp Pro Pro Lys Gly Asp
210 215 220
Pro Trp Leu Ala Val Leu Ser Ser Arg Asp Lys Lys Asp Ala Lys Tyr
225 230 235 240
Asp Asn Lys Leu Ile Ala Glu Ala Thr Lys Trp Met Lys Ala Leu Asn
245 250 255
Met Asn Gly Lys
260
<210> 80
<211> 299
<212> PRT
<213> 人工
<220>
<223> P4A + 信号肽
<400> 80
Met Ile Gln Lys Arg Lys Arg Thr Val Ser Phe Arg Leu Val Leu Met
1 5 10 15
Cys Thr Leu Leu Phe Val Ser Leu Pro Ile Thr Lys Thr Ser Ala Gln
20 25 30
Ala Ser Lys Thr Glu Met Lys Asp Asp Phe Ala Lys Leu Glu Glu Gln
35 40 45
Phe Asp Ala Lys Leu Gly Ile Phe Ala Leu Asp Thr Gly Thr Asn Arg
50 55 60
Thr Val Ala Tyr Arg Pro Asp Glu Arg Phe Ala Phe Ala Ser Thr Ile
65 70 75 80
Lys Ala Leu Thr Val Gly Val Leu Leu Gln Gln Lys Ser Ile Glu Asp
85 90 95
Leu Asn Gln Arg Ile Thr Tyr Thr Arg Asp Asp Leu Val Asn Tyr Asn
100 105 110
Pro Ile Thr Glu Lys His Val Asp Thr Gly Met Thr Leu Lys Glu Leu
115 120 125
Ala Asp Ala Ser Leu Arg Tyr Ser Asp Asn Ala Ala Gln Asn Leu Ile
130 135 140
Leu Lys Gln Ile Gly Gly Pro Glu Ser Leu Lys Lys Glu Leu Arg Lys
145 150 155 160
Ile Gly Asp Glu Val Thr Asn Pro Glu Arg Phe Glu Pro Glu Leu Asn
165 170 175
Glu Val Asn Pro Gly Glu Thr Gln Asp Thr Ser Thr Ala Arg Ala Leu
180 185 190
Val Thr Ser Leu Arg Ala Phe Ala Leu Glu Asp Lys Leu Pro Ser Glu
195 200 205
Lys Arg Glu Leu Leu Ile Asp Trp Met Lys Arg Asn Thr Thr Gly Asp
210 215 220
Ala Leu Ile Arg Ala Gly Val Pro Asp Gly Trp Glu Val Gly Asp Lys
225 230 235 240
Thr Gly Ser Gly Asp Tyr Gly Thr Arg Asn Asp Ile Ala Ile Ile Trp
245 250 255
Pro Pro Lys Gly Asp Pro Val Val Leu Ala Val Leu Ser Ser Arg Asp
260 265 270
Lys Lys Asp Ala Lys Tyr Asp Asn Lys Leu Ile Ala Glu Ala Thr Lys
275 280 285
Val Val Met Lys Ala Leu Asn Met Asn Gly Lys
290 295
<210> 81
<211> 263
<212> PRT
<213> 大肠埃希氏杆菌（Escherichia coli）
<400> 81
His Pro Glu Thr Leu Val Lys Val Lys Asp Ala Glu Asp Gln Leu Gly
1 5 10 15
Ala Arg Val Gly Tyr Ile Glu Leu Asp Leu Asn Ser Gly Lys Ile Leu
20 25 30
Glu Ser Phe Arg Pro Glu Glu Arg Phe Pro Met Val Ser Thr Phe Lys
35 40 45
Val Leu Leu Cys Gly Ala Val Leu Ser Arg Val Asp Ala Gly Gln Glu
50 55 60
Gln Leu Gly Arg Arg Ile His Tyr Ser Gln Asn Asp Leu Val Glu Tyr
65 70 75 80
Ser Pro Val Thr Glu Lys His Leu Thr Asp Gly Met Thr Val Arg Glu
85 90 95
Leu Cys Ser Ala Ala Ile Thr Met Ser Asp Asn Thr Ala Ala Asn Leu
100 105 110
Leu Leu Thr Thr Ile Gly Gly Pro Lys Glu Leu Thr Ala Phe Leu His
115 120 125
Asn Met Gly Asp His Val Thr Arg Leu Asp Arg Trp Glu Pro Glu Leu
130 135 140
Asn Glu Ala Ile Pro Asn Asp Glu Arg Asp Thr Thr Met Pro Ala Ala
145 150 155 160
Met Ala Thr Thr Leu Arg Lys Leu Leu Thr Gly Glu Leu Leu Thr Leu
165 170 175
Ala Ser Arg Gln Gln Leu Ile Asp Trp Met Glu Ala Asp Lys Val Ala
180 185 190
Gly Pro Leu Leu Arg Ser Ala Leu Pro Ala Gly Trp Phe Ile Ala Asp
195 200 205
Lys Ser Gly Ala Gly Glu Arg Gly Ser Arg Gly Ile Ile Ala Ala Leu
210 215 220
Gly Pro Asp Gly Lys Pro Ser Arg Ile Val Val Ile Tyr Thr Thr Gly
225 230 235 240
Ser Gln Ala Thr Met Asp Glu Arg Asp Arg Gln Ile Ala Glu Ile Gly
245 250 255
Ala Ser Leu Ile Lys His Trp
260
<210> 82
<211> 263
<212> PRT
<213> 大肠埃希氏杆菌（Escherichia coli）
<400> 82
Gln Thr Ser Ala Val Gln Gln Lys Leu Ala Ala Leu Glu Lys Ser Ser
1 5 10 15
Gly Gly Arg Leu Gly Val Ala Leu Ile Asp Thr Ala Asp Asn Thr Gln
20 25 30
Val Leu Tyr Arg Gly Asp Glu Arg Phe Pro Met Cys Ser Thr Ser Lys
35 40 45
Val Met Ala Ala Ala Ala Val Leu Lys Gln Ser Glu Thr Gln Lys Gln
50 55 60
Leu Leu Asn Gln Pro Val Glu Ile Lys Pro Ala Asp Leu Val Asn Tyr
65 70 75 80
Asn Pro Ile Ala Glu Lys His Val Asn Gly Thr Met Thr Leu Ala Glu
85 90 95
Leu Ser Ala Ala Ala Leu Gln Tyr Ser Asp Asn Thr Ala Met Asn Lys
100 105 110
Leu Ile Ala Gln Leu Gly Gly Pro Gly Gly Val Thr Ala Phe Ala Arg
115 120 125
Ala Ile Gly Asp Glu Thr Phe Arg Leu Asp Arg Thr Glu Pro Thr Leu
130 135 140
Asn Thr Ala Ile Pro Gly Asp Pro Arg Asp Thr Thr Thr Pro Arg Ala
145 150 155 160
Met Ala Gln Thr Leu Arg Gln Leu Thr Leu Gly His Ala Leu Gly Glu
165 170 175
Thr Gln Arg Ala Gln Leu Val Thr Trp Leu Lys Gly Asn Thr Thr Gly
180 185 190
Ala Ala Ser Ile Arg Ala Gly Leu Pro Thr Ser Trp Thr Ala Gly Asp
195 200 205
Lys Thr Gly Ser Gly Gly Tyr Gly Thr Thr Asn Asp Ile Ala Val Ile
210 215 220
Trp Pro Gln Gly Arg Ala Pro Leu Val Leu Val Thr Tyr Phe Thr Gln
225 230 235 240
Pro Gln Gln Asn Ala Glu Ser Arg Arg Asp Val Leu Ala Ser Ala Ala
245 250 255
Arg Ile Ile Ala Glu Gly Leu
260
<210> 83
<211> 257
<212> PRT
<213> 金黄色葡萄球菌（Staphylococcus aureus）
<400> 83
Lys Glu Leu Asn Asp Leu Glu Lys Lys Tyr Asn Ala His Ile Gly Val
1 5 10 15
Tyr Ala Leu Asp Thr Lys Ser Gly Lys Glu Val Lys Phe Asn Ser Asp
20 25 30
Lys Arg Phe Ala Tyr Ala Ser Thr Ser Lys Ala Ile Asn Ser Ala Ile
35 40 45
Leu Leu Glu Gln Val Pro Tyr Asn Lys Leu Asn Lys Lys Val His Ile
50 55 60
Asn Lys Asp Asp Ile Val Ala Tyr Ser Pro Ile Leu Glu Lys Tyr Val
65 70 75 80
Gly Lys Asp Ile Thr Leu Lys Ala Leu Ile Glu Ala Ser Met Thr Tyr
85 90 95
Ser Asp Asn Thr Ala Asn Asn Lys Ile Ile Lys Glu Ile Gly Gly Ile
100 105 110
Lys Lys Val Lys Gln Arg Leu Lys Glu Leu Gly Asp Lys Val Thr Asn
115 120 125
Pro Val Arg Tyr Glu Ile Glu Leu Asn Tyr Tyr Ser Pro Lys Ser Lys
130 135 140
Lys Asp Thr Ser Thr Pro Ala Ala Phe Gly Lys Thr Leu Asn Lys Leu
145 150 155 160
Ile Ala Asn Gly Lys Leu Ser Lys Glu Asn Lys Lys Phe Leu Leu Asp
165 170 175
Leu Met Leu Asn Asn Lys Ser Gly Asp Thr Leu Ile Lys Asp Gly Val
180 185 190
Pro Lys Asp Tyr Lys Val Ala Asp Lys Ser Gly Gln Ala Ile Thr Tyr
195 200 205
Ala Ser Arg Asn Asp Val Ala Phe Val Tyr Pro Lys Gly Gln Ser Glu
210 215 220
Pro Ile Val Leu Val Ile Phe Thr Asn Lys Asp Asn Lys Ser Asp Lys
225 230 235 240
Pro Asn Asp Lys Leu Ile Ser Glu Thr Ala Lys Ser Val Met Lys Glu
245 250 255
Phe
<210> 84
<211> 199
<212> PRT
<213> 大肠埃希氏杆菌（Escherichia coli）
<400> 84
Met Ser Asn Ala Lys Thr Lys Leu Gly Ile Thr Lys Tyr Ser Ile Val
1 5 10 15
Thr Asn Ser Asn Asp Ser Val Thr Leu Arg Leu Met Thr Glu His Asp
20 25 30
Leu Ala Met Leu Tyr Glu Trp Leu Asn Arg Ser His Ile Val Glu Trp
35 40 45
Trp Gly Gly Glu Glu Ala Arg Pro Thr Leu Ala Asp Val Gln Glu Gln
50 55 60
Tyr Leu Pro Ser Val Leu Ala Gln Glu Ser Val Thr Pro Tyr Ile Ala
65 70 75 80
Met Leu Asn Gly Glu Pro Ile Gly Tyr Ala Gln Ser Tyr Val Ala Leu
85 90 95
Gly Ser Gly Asp Gly Arg Trp Glu Glu Glu Thr Asp Pro Gly Val Arg
100 105 110
Gly Ile Asp Gln Leu Leu Ala Asn Ala Ser Gln Leu Gly Lys Gly Leu
115 120 125
Gly Thr Lys Leu Val Arg Ala Leu Val Glu Leu Leu Phe Asn Asp Pro
130 135 140
Glu Val Thr Lys Ile Gln Thr Asp Pro Ser Pro Ser Asn Leu Arg Ala
145 150 155 160
Ile Arg Cys Tyr Glu Lys Ala Gly Phe Glu Arg Gln Gly Thr Val Thr
165 170 175
Thr Pro Tyr Gly Pro Ala Val Tyr Met Val Gln Thr Arg Gln Ala Phe
180 185 190
Glu Arg Thr Arg Ser Asp Ala
195
<210> 85
<211> 419
<212> PRT
<213> 大肠埃希氏杆菌（Escherichia coli）
<400> 85
Met Arg Phe Glu Glu Trp Val Lys Asp Lys His Ile Pro Phe Lys Leu
1 5 10 15
Asn His Pro Asp Asp Asn Tyr Asp Asp Phe Lys Pro Leu Arg Lys Ile
20 25 30
Ile Gly Asp Thr Arg Val Val Ala Leu Gly Glu Asn Ser His Phe Ile
35 40 45
Lys Glu Phe Phe Leu Leu Arg His Thr Leu Leu Arg Phe Phe Ile Glu
50 55 60
Asp Leu Gly Phe Thr Thr Phe Ala Phe Glu Phe Gly Phe Ala Glu Gly
65 70 75 80
Gln Ile Ile Asn Asn Trp Ile His Gly Gln Gly Thr Asp Asp Glu Ile
85 90 95
Gly Arg Phe Leu Lys His Phe Tyr Tyr Pro Glu Glu Leu Lys Thr Thr
100 105 110
Phe Leu Trp Leu Arg Glu Tyr Asn Lys Ala Ala Lys Glu Lys Ile Thr
115 120 125
Phe Leu Gly Ile Asp Ile Pro Arg Asn Gly Gly Ser Tyr Leu Pro Asn
130 135 140
Met Glu Ile Val His Asp Phe Phe Arg Thr Ala Asp Lys Glu Ala Leu
145 150 155 160
His Ile Ile Asp Asp Ala Phe Asn Ile Ala Lys Lys Ile Asp Tyr Phe
165 170 175
Ser Thr Ser Gln Ala Ala Leu Asn Leu His Glu Leu Thr Asp Ser Glu
180 185 190
Lys Cys Arg Leu Thr Ser Gln Leu Ala Arg Val Lys Val Arg Leu Glu
195 200 205
Ala Met Ala Pro Ile His Ile Glu Lys Tyr Gly Ile Asp Lys Tyr Glu
210 215 220
Thr Ile Leu His Tyr Ala Asn Gly Met Ile Tyr Leu Asp Tyr Asn Ile
225 230 235 240
Gln Ala Met Ser Gly Phe Ile Ser Gly Gly Gly Met Gln Gly Asp Met
245 250 255
Gly Ala Lys Asp Lys Tyr Met Ala Asp Ser Val Leu Trp His Leu Lys
260 265 270
Asn Pro Gln Ser Glu Gln Lys Val Ile Val Val Ala His Asn Ala His
275 280 285
Ile Gln Lys Thr Pro Ile Leu Tyr Asp Gly Phe Leu Ser Cys Leu Pro
290 295 300
Met Gly Gln Arg Leu Lys Asn Ala Ile Gly Asp Asp Tyr Met Ser Leu
305 310 315 320
Gly Ile Thr Ser Tyr Ser Gly His Thr Ala Ala Leu Tyr Pro Glu Val
325 330 335
Asp Thr Lys Tyr Gly Phe Arg Val Asp Asn Phe Gln Leu Gln Glu Pro
340 345 350
Asn Glu Gly Ser Val Glu Lys Ala Ile Ser Gly Cys Gly Val Thr Asn
355 360 365
Ser Phe Val Phe Phe Arg Asn Ile Pro Glu Asp Leu Gln Ser Ile Pro
370 375 380
Asn Met Ile Arg Phe Asp Ser Ile Tyr Met Lys Ala Glu Leu Glu Lys
385 390 395 400
Ala Phe Asp Gly Ile Phe Gln Ile Glu Lys Ser Ser Val Ser Glu Val
405 410 415
Val Tyr Glu
<210> 86
<211> 388
<212> PRT
<213> 脆弱拟杆菌（Bacteroides fragilis）
<400> 86
Met Thr Met Arg Ile Asp Thr Asp Lys Gln Met Asn Leu Leu Ser Asp
1 5 10 15
Lys Asn Val Ala Ile Ile Gly Gly Gly Pro Val Gly Leu Thr Met Ala
20 25 30
Lys Leu Leu Gln Gln Asn Gly Ile Asp Val Ser Val Tyr Glu Arg Asp
35 40 45
Asn Asp Arg Glu Ala Arg Ile Phe Gly Gly Thr Leu Asp Leu His Lys
50 55 60
Gly Ser Gly Gln Glu Ala Met Lys Lys Ala Gly Leu Leu Gln Thr Tyr
65 70 75 80
Tyr Asp Leu Ala Leu Pro Met Gly Val Asn Ile Ala Asp Lys Lys Gly
85 90 95
Asn Ile Leu Ser Thr Lys Asn Val Lys Pro Glu Asn Arg Phe Asp Asn
100 105 110
Pro Glu Ile Asn Arg Asn Asp Leu Arg Ala Ile Leu Leu Asn Ser Leu
115 120 125
Glu Asn Asp Thr Val Ile Trp Asp Arg Lys Leu Val Met Leu Glu Pro
130 135 140
Gly Lys Lys Lys Trp Thr Leu Thr Phe Glu Asn Lys Pro Ser Glu Thr
145 150 155 160
Ala Asp Leu Val Ile Leu Ala Asn Gly Gly Met Ser Lys Val Arg Lys
165 170 175
Phe Val Thr Asp Thr Glu Val Glu Glu Thr Gly Thr Phe Asn Ile Gln
180 185 190
Ala Asp Ile His Gln Pro Glu Ile Asn Cys Pro Gly Phe Phe Gln Leu
195 200 205
Cys Asn Gly Asn Arg Leu Met Ala Ser His Gln Gly Asn Leu Leu Phe
210 215 220
Ala Asn Pro Asn Asn Asn Gly Ala Leu His Phe Gly Ile Ser Phe Lys
225 230 235 240
Thr Pro Asp Glu Trp Lys Asn Gln Thr Gln Val Asp Phe Gln Asn Arg
245 250 255
Asn Ser Val Val Asp Phe Leu Leu Lys Glu Phe Ser Asp Trp Asp Glu
260 265 270
Arg Tyr Lys Glu Leu Ile His Thr Thr Leu Ser Phe Val Gly Leu Ala
275 280 285
Thr Arg Ile Phe Pro Leu Glu Lys Pro Trp Lys Ser Lys Arg Pro Leu
290 295 300
Pro Ile Thr Met Ile Gly Asp Ala Ala His Leu Met Pro Pro Phe Ala
305 310 315 320
Gly Gln Gly Val Asn Ser Gly Leu Val Asp Ala Leu Ile Leu Ser Asp
325 330 335
Asn Leu Ala Asp Gly Lys Phe Asn Ser Ile Glu Glu Ala Val Lys Asn
340 345 350
Tyr Glu Gln Gln Met Phe Met Tyr Gly Lys Glu Ala Gln Glu Glu Ser
355 360 365
Thr Gln Asn Glu Ile Glu Met Phe Lys Pro Asp Phe Thr Phe Gln Gln
370 375 380
Leu Leu Asn Val
385
<210> 87
<211> 267
<212> PRT
<213> 屎肠球菌（Enterococcus faecium）
<400> 87
Met Leu Lys Gln Lys Glu Leu Ile Ala Asn Val Lys Asn Leu Thr Glu
1 5 10 15
Ser Asp Glu Arg Ile Thr Ala Cys Met Met Tyr Gly Ser Phe Thr Lys
20 25 30
Gly Glu Gly Asp Gln Tyr Ser Asp Ile Glu Phe Tyr Ile Phe Leu Lys
35 40 45
His Ser Ile Thr Ser Asn Phe Asp Ser Ser Asn Trp Leu Phe Asp Val
50 55 60
Ala Pro Tyr Leu Met Leu Tyr Lys Asn Glu Tyr Gly Thr Glu Val Val
65 70 75 80
Ile Phe Asp Asn Leu Ile Arg Gly Glu Phe His Phe Leu Ser Glu Lys
85 90 95
Asp Met Asn Ile Ile Pro Ser Phe Lys Asp Ser Gly Tyr Ile Pro Asp
100 105 110
Thr Lys Ala Met Leu Ile Tyr Asp Glu Thr Gly Gln Leu Glu Asn Tyr
115 120 125
Leu Ser Glu Ile Ser Gly Ala Arg Pro Asn Arg Leu Thr Glu Glu Asn
130 135 140
Ala Asn Phe Leu Leu Cys Asn Phe Ser Asn Leu Trp Leu Met Gly Ile
145 150 155 160
Asn Val Leu Lys Arg Gly Glu Tyr Ala Arg Ser Leu Glu Leu Leu Ser
165 170 175
Gln Leu Gln Lys Asn Thr Leu Gln Leu Ile Arg Met Ala Glu Lys Asn
180 185 190
Ala Asp Asn Trp Leu Asn Met Ser Lys Asn Leu Glu Lys Glu Ile Ser
195 200 205
Leu Glu Asn Tyr Lys Lys Phe Ala Lys Thr Thr Ala Arg Leu Asp Lys
210 215 220
Val Glu Leu Phe Glu Ala Tyr Lys Asn Ser Leu Leu Leu Val Met Asp
225 230 235 240
Leu Gln Ser His Leu Ile Glu Gln Tyr Asn Leu Lys Val Thr His Asp
245 250 255
Ile Leu Glu Arg Leu Leu Asn Tyr Ile Ser Glu
260 265
<210> 88
<211> 532
<212> PRT
<213> 人工
<220>
<223> TEM36-GGGGGG-CTXM16
<400> 88
His Pro Glu Thr Leu Val Lys Val Lys Asp Ala Glu Asp Gln Leu Gly
1 5 10 15
Ala Arg Val Gly Tyr Ile Glu Leu Asp Leu Asn Ser Gly Lys Ile Leu
20 25 30
Glu Ser Phe Arg Pro Glu Glu Arg Phe Pro Met Val Ser Thr Phe Lys
35 40 45
Val Leu Leu Cys Gly Ala Val Leu Ser Arg Val Asp Ala Gly Gln Glu
50 55 60
Gln Leu Gly Arg Arg Ile His Tyr Ser Gln Asn Asp Leu Val Glu Tyr
65 70 75 80
Ser Pro Val Thr Glu Lys His Leu Thr Asp Gly Met Thr Val Arg Glu
85 90 95
Leu Cys Ser Ala Ala Ile Thr Met Ser Asp Asn Thr Ala Ala Asn Leu
100 105 110
Leu Leu Thr Thr Ile Gly Gly Pro Lys Glu Leu Thr Ala Phe Leu His
115 120 125
Asn Met Gly Asp His Val Thr Arg Leu Asp Arg Trp Glu Pro Glu Leu
130 135 140
Asn Glu Ala Ile Pro Asn Asp Glu Arg Asp Thr Thr Met Pro Ala Ala
145 150 155 160
Met Ala Thr Thr Leu Arg Lys Leu Leu Thr Gly Glu Leu Leu Thr Leu
165 170 175
Ala Ser Arg Gln Gln Leu Ile Asp Trp Met Glu Ala Asp Lys Val Ala
180 185 190
Gly Pro Leu Leu Arg Ser Ala Leu Pro Ala Gly Trp Phe Ile Ala Asp
195 200 205
Lys Ser Gly Ala Gly Glu Arg Gly Ser Arg Gly Ile Ile Ala Ala Leu
210 215 220
Gly Pro Asp Gly Lys Pro Ser Arg Ile Val Val Ile Tyr Thr Thr Gly
225 230 235 240
Ser Gln Ala Thr Met Asp Glu Arg Asp Arg Gln Ile Ala Glu Ile Gly
245 250 255
Ala Ser Leu Ile Lys His Trp Gly Gly Gly Gly Gly Gly Gln Thr Ser
260 265 270
Ala Val Gln Gln Lys Leu Ala Ala Leu Glu Lys Ser Ser Gly Gly Arg
275 280 285
Leu Gly Val Ala Leu Ile Asp Thr Ala Asp Asn Thr Gln Val Leu Tyr
290 295 300
Arg Gly Asp Glu Arg Phe Pro Met Cys Ser Thr Ser Lys Val Met Ala
305 310 315 320
Ala Ala Ala Val Leu Lys Gln Ser Glu Thr Gln Lys Gln Leu Leu Asn
325 330 335
Gln Pro Val Glu Ile Lys Pro Ala Asp Leu Val Asn Tyr Asn Pro Ile
340 345 350
Ala Glu Lys His Val Asn Gly Thr Met Thr Leu Ala Glu Leu Ser Ala
355 360 365
Ala Ala Leu Gln Tyr Ser Asp Asn Thr Ala Met Asn Lys Leu Ile Ala
370 375 380
Gln Leu Gly Gly Pro Gly Gly Val Thr Ala Phe Ala Arg Ala Ile Gly
385 390 395 400
Asp Glu Thr Phe Arg Leu Asp Arg Thr Glu Pro Thr Leu Asn Thr Ala
405 410 415
Ile Pro Gly Asp Pro Arg Asp Thr Thr Thr Pro Arg Ala Met Ala Gln
420 425 430
Thr Leu Arg Gln Leu Thr Leu Gly His Ala Leu Gly Glu Thr Gln Arg
435 440 445
Ala Gln Leu Val Thr Trp Leu Lys Gly Asn Thr Thr Gly Ala Ala Ser
450 455 460
Ile Arg Ala Gly Leu Pro Thr Ser Trp Thr Ala Gly Asp Lys Thr Gly
465 470 475 480
Ser Gly Gly Tyr Gly Thr Thr Asn Asp Ile Ala Val Ile Trp Pro Gln
485 490 495
Gly Arg Ala Pro Leu Val Leu Val Thr Tyr Phe Thr Gln Pro Gln Gln
500 505 510
Asn Ala Glu Ser Arg Arg Asp Val Leu Ala Ser Ala Ala Arg Ile Ile
515 520 525
Ala Glu Gly Leu
530
<210> 89
<211> 532
<212> PRT
<213> 人工
<220>
<223> 融合体CTXM16-6G-TEM36
<400> 89
Gln Thr Ser Ala Val Gln Gln Lys Leu Ala Ala Leu Glu Lys Ser Ser
1 5 10 15
Gly Gly Arg Leu Gly Val Ala Leu Ile Asp Thr Ala Asp Asn Thr Gln
20 25 30
Val Leu Tyr Arg Gly Asp Glu Arg Phe Pro Met Cys Ser Thr Ser Lys
35 40 45
Val Met Ala Ala Ala Ala Val Leu Lys Gln Ser Glu Thr Gln Lys Gln
50 55 60
Leu Leu Asn Gln Pro Val Glu Ile Lys Pro Ala Asp Leu Val Asn Tyr
65 70 75 80
Asn Pro Ile Ala Glu Lys His Val Asn Gly Thr Met Thr Leu Ala Glu
85 90 95
Leu Ser Ala Ala Ala Leu Gln Tyr Ser Asp Asn Thr Ala Met Asn Lys
100 105 110
Leu Ile Ala Gln Leu Gly Gly Pro Gly Gly Val Thr Ala Phe Ala Arg
115 120 125
Ala Ile Gly Asp Glu Thr Phe Arg Leu Asp Arg Thr Glu Pro Thr Leu
130 135 140
Asn Thr Ala Ile Pro Gly Asp Pro Arg Asp Thr Thr Thr Pro Arg Ala
145 150 155 160
Met Ala Gln Thr Leu Arg Gln Leu Thr Leu Gly His Ala Leu Gly Glu
165 170 175
Thr Gln Arg Ala Gln Leu Val Thr Trp Leu Lys Gly Asn Thr Thr Gly
180 185 190
Ala Ala Ser Ile Arg Ala Gly Leu Pro Thr Ser Trp Thr Ala Gly Asp
195 200 205
Lys Thr Gly Ser Gly Gly Tyr Gly Thr Thr Asn Asp Ile Ala Val Ile
210 215 220
Trp Pro Gln Gly Arg Ala Pro Leu Val Leu Val Thr Tyr Phe Thr Gln
225 230 235 240
Pro Gln Gln Asn Ala Glu Ser Arg Arg Asp Val Leu Ala Ser Ala Ala
245 250 255
Arg Ile Ile Ala Glu Gly Leu Gly Gly Gly Gly Gly Gly His Pro Glu
260 265 270
Thr Leu Val Lys Val Lys Asp Ala Glu Asp Gln Leu Gly Ala Arg Val
275 280 285
Gly Tyr Ile Glu Leu Asp Leu Asn Ser Gly Lys Ile Leu Glu Ser Phe
290 295 300
Arg Pro Glu Glu Arg Phe Pro Met Val Ser Thr Phe Lys Val Leu Leu
305 310 315 320
Cys Gly Ala Val Leu Ser Arg Val Asp Ala Gly Gln Glu Gln Leu Gly
325 330 335
Arg Arg Ile His Tyr Ser Gln Asn Asp Leu Val Glu Tyr Ser Pro Val
340 345 350
Thr Glu Lys His Leu Thr Asp Gly Met Thr Val Arg Glu Leu Cys Ser
355 360 365
Ala Ala Ile Thr Met Ser Asp Asn Thr Ala Ala Asn Leu Leu Leu Thr
370 375 380
Thr Ile Gly Gly Pro Lys Glu Leu Thr Ala Phe Leu His Asn Met Gly
385 390 395 400
Asp His Val Thr Arg Leu Asp Arg Trp Glu Pro Glu Leu Asn Glu Ala
405 410 415
Ile Pro Asn Asp Glu Arg Asp Thr Thr Met Pro Ala Ala Met Ala Thr
420 425 430
Thr Leu Arg Lys Leu Leu Thr Gly Glu Leu Leu Thr Leu Ala Ser Arg
435 440 445
Gln Gln Leu Ile Asp Trp Met Glu Ala Asp Lys Val Ala Gly Pro Leu
450 455 460
Leu Arg Ser Ala Leu Pro Ala Gly Trp Phe Ile Ala Asp Lys Ser Gly
465 470 475 480
Ala Gly Glu Arg Gly Ser Arg Gly Ile Ile Ala Ala Leu Gly Pro Asp
485 490 495
Gly Lys Pro Ser Arg Ile Val Val Ile Tyr Thr Thr Gly Ser Gln Ala
500 505 510
Thr Met Asp Glu Arg Asp Arg Gln Ile Ala Glu Ile Gly Ala Ser Leu
515 520 525
Ile Lys His Trp
530
<210> 90
<211> 537
<212> PRT
<213> 人工
<220>
<223> 融合体TEM36 - G(EAAAK)2 - CTXM16
<400> 90
His Pro Glu Thr Leu Val Lys Val Lys Asp Ala Glu Asp Gln Leu Gly
1 5 10 15
Ala Arg Val Gly Tyr Ile Glu Leu Asp Leu Asn Ser Gly Lys Ile Leu
20 25 30
Glu Ser Phe Arg Pro Glu Glu Arg Phe Pro Met Val Ser Thr Phe Lys
35 40 45
Val Leu Leu Cys Gly Ala Val Leu Ser Arg Val Asp Ala Gly Gln Glu
50 55 60
Gln Leu Gly Arg Arg Ile His Tyr Ser Gln Asn Asp Leu Val Glu Tyr
65 70 75 80
Ser Pro Val Thr Glu Lys His Leu Thr Asp Gly Met Thr Val Arg Glu
85 90 95
Leu Cys Ser Ala Ala Ile Thr Met Ser Asp Asn Thr Ala Ala Asn Leu
100 105 110
Leu Leu Thr Thr Ile Gly Gly Pro Lys Glu Leu Thr Ala Phe Leu His
115 120 125
Asn Met Gly Asp His Val Thr Arg Leu Asp Arg Trp Glu Pro Glu Leu
130 135 140
Asn Glu Ala Ile Pro Asn Asp Glu Arg Asp Thr Thr Met Pro Ala Ala
145 150 155 160
Met Ala Thr Thr Leu Arg Lys Leu Leu Thr Gly Glu Leu Leu Thr Leu
165 170 175
Ala Ser Arg Gln Gln Leu Ile Asp Trp Met Glu Ala Asp Lys Val Ala
180 185 190
Gly Pro Leu Leu Arg Ser Ala Leu Pro Ala Gly Trp Phe Ile Ala Asp
195 200 205
Lys Ser Gly Ala Gly Glu Arg Gly Ser Arg Gly Ile Ile Ala Ala Leu
210 215 220
Gly Pro Asp Gly Lys Pro Ser Arg Ile Val Val Ile Tyr Thr Thr Gly
225 230 235 240
Ser Gln Ala Thr Met Asp Glu Arg Asp Arg Gln Ile Ala Glu Ile Gly
245 250 255
Ala Ser Leu Ile Lys His Trp Gly Glu Ala Ala Ala Lys Glu Ala Ala
260 265 270
Ala Lys Gln Thr Ser Ala Val Gln Gln Lys Leu Ala Ala Leu Glu Lys
275 280 285
Ser Ser Gly Gly Arg Leu Gly Val Ala Leu Ile Asp Thr Ala Asp Asn
290 295 300
Thr Gln Val Leu Tyr Arg Gly Asp Glu Arg Phe Pro Met Cys Ser Thr
305 310 315 320
Ser Lys Val Met Ala Ala Ala Ala Val Leu Lys Gln Ser Glu Thr Gln
325 330 335
Lys Gln Leu Leu Asn Gln Pro Val Glu Ile Lys Pro Ala Asp Leu Val
340 345 350
Asn Tyr Asn Pro Ile Ala Glu Lys His Val Asn Gly Thr Met Thr Leu
355 360 365
Ala Glu Leu Ser Ala Ala Ala Leu Gln Tyr Ser Asp Asn Thr Ala Met
370 375 380
Asn Lys Leu Ile Ala Gln Leu Gly Gly Pro Gly Gly Val Thr Ala Phe
385 390 395 400
Ala Arg Ala Ile Gly Asp Glu Thr Phe Arg Leu Asp Arg Thr Glu Pro
405 410 415
Thr Leu Asn Thr Ala Ile Pro Gly Asp Pro Arg Asp Thr Thr Thr Pro
420 425 430
Arg Ala Met Ala Gln Thr Leu Arg Gln Leu Thr Leu Gly His Ala Leu
435 440 445
Gly Glu Thr Gln Arg Ala Gln Leu Val Thr Trp Leu Lys Gly Asn Thr
450 455 460
Thr Gly Ala Ala Ser Ile Arg Ala Gly Leu Pro Thr Ser Trp Thr Ala
465 470 475 480
Gly Asp Lys Thr Gly Ser Gly Gly Tyr Gly Thr Thr Asn Asp Ile Ala
485 490 495
Val Ile Trp Pro Gln Gly Arg Ala Pro Leu Val Leu Val Thr Tyr Phe
500 505 510
Thr Gln Pro Gln Gln Asn Ala Glu Ser Arg Arg Asp Val Leu Ala Ser
515 520 525
Ala Ala Arg Ile Ile Ala Glu Gly Leu
530 535

Claims (14)

1.一种用于在需要抗癌治疗的对象中提高抗癌药剂效能的吸附剂，其中待治疗的对象接受、将接受或已接受引起生态失调的药剂。
2.根据权利要求1所述的用途的吸附剂，其中所述引起生态失调的药剂是被给药用于预防或治疗感染的抗生素。
3.根据权利要求1或2所述的用途的吸附剂，其中所述吸附剂是活性炭。
4.根据权利要求1至3中的任一项所述的用途的吸附剂，其中所述吸附剂在包含下述组分的制剂中：
-含有吸附剂的核心，和
-外部包衣层，所述外部包衣层围绕所述核心形成，使得所述吸附剂在肠的下部中从所述制剂释放。
5.根据权利要求4所述的用途的吸附剂，其中所述核心还包含卡拉胶，例如κ-卡拉胶。
6.一种用于在需要抗癌疗法的对象中提高抗癌药剂效能的方法中使用的抗生素失活酶，其中所述对象接受、将接受或已接受用于预防或治疗感染的抗生素。
7.根据权利要求6所述的用途的抗生素失活酶，其中(i)所述酶是β-内酰胺酶，特别是VIM-2或ribaxamase，并且所述抗生素是β-内酰胺类抗生素；或(ii)所述酶是是红霉素酯酶，并且所述抗生素是大环内酯。
8.根据权利要求6所述的用途的抗生素失活酶，其中所述酶是包含键合在一起的两种抗生素失活酶的杂合蛋白分子，所述两种酶使相同或不同的抗生素或来自于相同或不同类别的抗生素失活。
9.根据权利要求6至8中的任一项所述的用途的抗生素失活酶，其被配制在用于口服给药的组合物中，所述组合物适合于在肠的所需部分例如肠的下部中释放所述抗生素失活酶。
10.根据权利要求1至5中的任一项所述的用途的吸附剂或根据权利要求6至9中的任一项所述的用途的抗生素失活酶，其中所述抗癌药剂选自阿法替尼、阿柏西普、阿仑单抗、阿利维甲酸、六甲蜜胺、阿那格雷、三氧化二砷、天冬酰胺酶、阿特珠单抗、阿维单抗、阿西替尼、阿扎胞苷、苯达莫司汀、贝伐单抗、蓓萨罗丁、博来霉素、硼替佐米、博舒替尼、白消安、卡巴他赛、卡培他滨、卡铂、卡莫氟、卡莫司汀、西妥昔单抗、苯丁酸氮芥、盐酸氮芥、顺铂、克拉屈滨、氯法拉滨、克唑替尼、环磷酰胺、阿糖胞苷、达卡巴嗪、放线菌素D、达沙替尼、道诺霉素、地西他滨、地尼白介素、地诺单抗、多西他赛、多柔比星、德瓦鲁单抗、表柔比星、埃罗替尼、雌莫司汀、依托泊苷、依维莫司、氟尿苷、氟达拉滨、氟尿嘧啶、福莫司汀、吉非替尼、吉西他滨、吉妥单抗、羟基脲、替伊莫单抗、伊达比星、异环磷酰胺、伊马替尼、伊匹单抗、伊立替康、异维甲酸、伊沙匹隆、拉帕替尼、来那度胺、洛莫司汀、美法仑、巯基嘌呤、甲氨蝶呤、丝裂霉素、米托蒽醌、奈达铂、奈拉滨、尼罗替尼、纳武单抗、奥法木单抗、奥沙利铂、紫杉醇、帕尼单抗、帕比司他、帕唑帕尼、派姆单抗、培美曲塞、喷司他丁、帕妥珠单抗、泊马度胺、帕纳替尼、丙卡巴肼、雷替曲塞、瑞戈非尼、利妥昔单抗、罗米地辛、鲁索替尼、索拉非尼、链脲佐菌素、舒尼替尼、他米巴罗汀、替加氟、替莫唑胺、西罗莫司脂化物、替尼泊苷、沙利度胺、硫鸟嘌呤、拓扑替康、托西莫单抗、曲妥珠单抗、维甲酸、丙戊酸盐、戊柔比星、凡德他尼、维罗非尼、长春花碱、长春新碱、长春地辛、长春氟宁、长春瑞滨和伏立诺他。
11.根据权利要求1至10中的任一项所述的用途的吸附剂或抗生素失活酶，其中所述抗癌药剂是免疫肿瘤药剂。
12.根据权利要求11所述的用途的吸附剂或抗生素失活酶，其中所述免疫肿瘤药剂选自：
-免疫检查点抑制剂，例如
PD-1抑制剂，例如纳武单抗或派姆单抗；或
PD-L1抑制剂，例如阿特珠单抗、阿维单抗或德瓦鲁单抗；或
CTLA-4抑制剂，例如伊匹单抗；或
PD-L2抑制剂；
-单克隆抗体，例如曲妥珠单抗；
-癌症疫苗，例如西普鲁塞-T；
-非特异性免疫疗法，例如来那度胺、干扰素或白介素；以及
-嵌合抗原受体(CAR)-T细胞疗法，例如tisagenlecleucel或axicabtageneciloleucel。
13.根据权利要求1至11中的任一项所述的用途的吸附剂或抗生素失活酶，其中将所述抗癌药剂与至少一种其他抗癌药剂、特别是与至少一种免疫肿瘤药剂组合。
14.根据权利要求1至13中的任一项所述的用途的吸附剂或抗生素失活酶，其中所述癌症选自急性成淋巴细胞性白血病(ALL)、急性髓系白血病(AML)、肾上腺皮质癌、肛门癌、阑尾癌、非典型畸胎瘤/横纹肌样瘤、基底细胞癌、胆管癌、膀胱癌、骨癌、脑肿瘤、星形细胞瘤、脑和脊髓肿瘤、脑干神经胶质瘤、中枢神经系统非典型畸胎瘤/横纹肌样瘤、中枢神经系统胚胎性肿瘤、乳腺癌、支气管肿瘤、伯基特淋巴瘤、类癌瘤、原发灶不明的转移癌、中枢神经系统癌症、宫颈癌、儿童期癌症、脊索瘤、慢性淋巴细胞性白血病(CLL)、慢性髓系白血病(CML)、慢性骨髓增殖性障碍、结肠癌、结肠直肠癌、颅咽管瘤、皮肤T细胞淋巴瘤导管原位癌(DCIS)、胚胎性肿瘤、子宫内膜癌、成室管膜细胞瘤、室管膜瘤、食管癌、嗅神经母细胞瘤、尤文氏肉瘤、颅外生殖细胞瘤、性腺外生殖细胞瘤、肝外胆管癌、眼癌、骨纤维组织细胞瘤、胆囊癌、胃癌、胃肠类癌瘤、胃肠间质瘤(GIST)、生殖细胞瘤、颅外生殖细胞瘤、性腺外生殖细胞瘤、卵巢生殖细胞瘤、妊娠滋养细胞肿瘤、神经胶质瘤、毛细胞白血病、头颈癌、心癌、肝细胞癌、组织细胞增生症、朗格汉斯细胞癌、霍奇金淋巴瘤、下咽癌、眼内黑素瘤、胰岛细胞瘤、卡波斯肉瘤、肾癌、朗格汉斯细胞组织细胞增生症、喉癌、白血病、唇和口腔癌、肝癌、原位小叶癌(LCIS)、肺癌、淋巴瘤、AIDS相关淋巴瘤、巨球蛋白血症、男性乳腺癌、成神经管细胞瘤、髓上皮瘤、黑素瘤、梅克尔细胞癌、恶性间皮瘤、隐匿原发的转移性鳞状颈部癌、涉及NUT基因的中线导管癌、口腔癌、多发性内分泌肿瘤综合征、多发性骨髓瘤/浆细胞瘤、蕈样肉芽肿、骨髓增生异常综合征、骨髓增生异常/骨髓增殖性肿瘤、慢性髓系白血病(CML)、急性髓系白血病(AML)、骨髓瘤、多发性骨髓瘤、慢性骨髓增殖性障碍、鼻腔癌、副鼻窦癌、鼻咽癌、成神经细胞瘤、非霍奇金淋巴瘤、非小细胞肺癌、口癌、口腔癌、唇癌、口咽癌、骨肉瘤、卵巢癌、胰腺癌、乳头状瘤病、副神经节瘤、副鼻窦癌、鼻腔癌、甲状旁腺癌、阴茎癌、咽癌、嗜铬细胞瘤、中间分化的松果体实质肿瘤、松果体母细胞瘤、垂体瘤、浆细胞瘤、胸膜肺母细胞瘤、乳腺癌、原发性中枢神经系统(CNS)淋巴瘤、前列腺癌、直肠癌、肾细胞癌、透明细胞肾细胞癌、肾盂癌、输尿管癌、移行细胞癌、成视网膜细胞瘤、横纹肌肉瘤、唾液腺癌、肉瘤、塞扎里综合征、皮肤癌、小细胞肺癌、小肠癌、软组织肉瘤、鳞状细胞癌、隐匿原发的鳞状颈癌(例如转移性)、头颈部鳞状细胞癌(HNSCC)、胃癌、幕上原始神经外胚层肿瘤、T-细胞淋巴瘤、睾丸癌、喉癌、胸腺瘤、胸腺癌、甲状腺癌、肾盂和输尿管的移行细胞癌、三阴性乳腺癌(TBC)、原发灶不明的妊娠滋养细胞肿瘤、儿童期罕见癌症、尿道癌、子宫癌、子宫肉瘤、华氏巨球蛋白血症和威尔姆氏瘤。
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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002061019A1 (en) * 2001-02-01 2002-08-08 Seng-Ha Hoang Manufacturing method of pine charcoal and novel use thereof as an anticancer agent
CN1657100A (zh) * 2004-02-19 2005-08-24 中国人民解放军军事医学科学院毒物药物研究所 选择性靶向胃癌和淋巴系统的纳米级制剂及其制备方法
CN101128187A (zh) * 2005-02-09 2008-02-20 达沃尔泰拉公司 活性剂的结肠递送
CN102869351A (zh) * 2010-02-23 2013-01-09 达·沃尔泰拉公司 用于吸附剂在肠内的经口递送的制剂
WO2017003908A1 (en) * 2015-06-30 2017-01-05 The Trustees Of Columbia University In The City Of New York Talc-bound compositions and uses thereof

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2008540602A (ja) 2005-05-18 2008-11-20 ダ・ボルテラ 吸着剤の結腸送達
US8048413B2 (en) 2006-05-17 2011-11-01 Helene Huguet Site-specific intestinal delivery of adsorbents, alone or in combination with degrading molecules
FI119190B (fi) 2006-06-21 2008-08-29 Ipsat Therapies Oy Modifioitu beta-laktamaasi ja menetelmä sen valmistamiseksi
FI20105572A0 (fi) 2010-05-24 2010-05-24 Prevab R Lcc Muokattu beeta-laktamaasi ja siihen liittyvät menetelmät ja käytöt
CN104797243A (zh) 2012-09-21 2015-07-22 辉凌公司 药物组合物
EP3132033B1 (en) 2014-04-17 2019-10-30 Synthetic Biologics, Inc. Beta-lactamases with improved properties for therapy
FR3027307B1 (fr) 2014-10-16 2016-11-04 Azurrx Sas Molecule proteique hybride apte a inhiber au moins un antibiotique et composition pharmaceutique la comportant

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002061019A1 (en) * 2001-02-01 2002-08-08 Seng-Ha Hoang Manufacturing method of pine charcoal and novel use thereof as an anticancer agent
CN1657100A (zh) * 2004-02-19 2005-08-24 中国人民解放军军事医学科学院毒物药物研究所 选择性靶向胃癌和淋巴系统的纳米级制剂及其制备方法
CN101128187A (zh) * 2005-02-09 2008-02-20 达沃尔泰拉公司 活性剂的结肠递送
CN102869351A (zh) * 2010-02-23 2013-01-09 达·沃尔泰拉公司 用于吸附剂在肠内的经口递送的制剂
WO2017003908A1 (en) * 2015-06-30 2017-01-05 The Trustees Of Columbia University In The City Of New York Talc-bound compositions and uses thereof

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