申请/专利权人：百时美施贵宝公司

申请日：2015-10-23

公开（公告）日：2024-03-15

公开（公告）号：CN110078814B

主分类号：C07K14/50

分类号：C07K14/50;C12N15/12;A61K45/06;A61K47/60;A61P3/10;A61P3/04;A61P1/16;A61P9/04;A61P9/00;A61P13/12;A61P11/00

优先权：["20141024 US 62/068,523","20141024 US 62/068,514","20141024 US 62/068,526","20141024 US 62/068,534","20141024 US 62/068,296","20150401 US 62/141,337","20150401 US 62/141,383"]

专利状态码：有效-授权

法律状态：2024.03.15#授权;2019.08.27#实质审查的生效;2019.08.02#公开

摘要：提供了修饰的FGF‑21多肽及其用途，例如用于治疗纤维化相关疾病。公开了含有内部缺失及任选的置换肽的修饰的FGF‑21多肽，其任选地经至少一个非天然编码的氨基酸修饰和或任选地融合至融合配偶体。

主权项：1.修饰的FGF-21多肽在制备用于治疗纤维化相关疾病的药物中的用途，所述纤维化相关疾病选自非酒精性脂肪性肝炎NASH、肝纤维化或肝硬化，其中所述修饰的FGF-21多肽由SEQIDNO:201的多肽组成，其中其对-乙酰基-苯丙氨酸残基连接至聚乙二醇部分。

全文数据：修饰的FGF-21多肽及其用途本申请为申请日为2015年10月23日、申请号为201580070276.X、发明名称为“修饰的FGF-21多肽及其用途”的发明专利申请的分案申请。发明领域本发明涉及含有内部缺失的任选地经肽置换的修饰的FGF-21多肽及其用于治疗或预防疾病和病症的用途。发明背景成纤维细胞生长因子是广泛表达于发育和成熟组织中的多肽Baird等，CancerCells,3:239-243,1991，其在多个生理功能中起关键作用McKeehan等，Prog.NucleicAcidRes.Mol.Biol.59:135-176,1998；Burgess,W.H.等，Annu.Rev.Biochem.58:575-6061989。根据文献，FGF家族由至少二十二个成员组成Reuss等，CellTissueRes.313:139-1572003。成纤维细胞生长因子21FGF-21已描述于文献中Nishimura等，BiochimicaetBiophysicaActa,1492:203-2062000；WO0136640；和WO0118172及美国专利公开No.20040259780，其各自通过提述以其整体并入本文。不同于其他FGF，已报导FGF-21不具有增殖及致瘤效应Ornitz和Itoh,GenomeBiology2001,23:评述3005.1-3005.12。一些FGF-21多肽及其用途描述于美国专利公开No.20010012628、美国专利No.6,716,626,U.S、美国专利公开No.20040259780、WO03011213、Kharitonenkov等JClinInvest.2005年6月；1156:1627-35；WO03059270；美国专利公开No.20050176631，WO2005091944；WO20070293430；美国专利公开No.20070293430：WO2008121563；美国专利No.4,904,584；WO9967291；WO9903887；WO0026354及美国专利No.5,218,092中，其各自通过提述以其整体并入本文。已报导人FGF-21在体内倾向于经历蛋白水解，在体外形成聚集体，经历脱酰胺Gimeno和Moller,TrendsEndocrinolMetab.2014Jun；256:303-11；美国专利No.8,361,963；Hecht等，PLoSOne.2012；711:e49345；美国专利公开No.20070293430；WO20060065582，有可能限制含有FGF-21的药物组合物的储存期限。聚集体及脱酰胺形式的治疗性多肽可潜在地增加免疫原性参见美国卫生及公共服务部U.S.DepartmentofHealthandHumanServices,“ImmunogenicityAssessmentforTherapeuticProteinProducts”,2014年8月；Subramanyam编，“TherapeuticProteinImmunogenicityFocusGroupNewsletter”,AmericanAssociationofPharmaceuticalScientists，第1卷，第3期2011年12月。以WO2008121563和美国专利公开No.20080255045公开的先前工作指示，经修饰以在特定位置含有连接至聚乙二醇的非天然编码的氨基酸的一些人FGF-21多肽展现出增加的体内半衰期和或保留的生物活性。然而，例示的人FGF-21多肽不包括本文所述的序列缺失或取代。纤维化是在器官或组织中过量纤维结缔组织的形成。纤维组织的过量沉积与可导致器官或组织功能损伤的病理性条件相关。受影响器官可包括肺肺纤维化lungpulmonaryfibrosis、肝脏肝纤维化liverhepaticfibrosis、肾脏肾纤维化kidneyrenalfibrosis及心脏心脏纤维化。纤维化还可影响其他组织及器官，包括关节、皮肤、肠、骨髓及其他组织及器官。例示性纤维化病况或疾病包括但不限于非酒精性脂肪性肝炎NASH，其影响肝脏；糖尿病性肾脏疾病及糖尿病性肾病变，其影响肾脏；及代谢性心脏衰竭，其影响心脏。例如，NASH的特征在于食用极少或不食用酒精的人的肝脏中的脂肪、炎症和损伤且可导致肝硬化。NASH倾向于在通常具有升高的血脂水平和患糖尿病或前驱糖尿病的超重或肥胖中年人中诊断出来。本发明的实施方案此外还阐述了与FGF-21多肽的活性和生产相关的问题，具有改进的生物学或药理学特性如改进的治疗性半衰期的FGF-21多肽的生产，和治疗或预防疾病和病症的方法。发明内容本文提供修饰的FGF-21多肽，其包含具有选自SEQIDNO:1-7的氨基酸序列的多肽，只是其中所述氨基酸序列包含：i2至19个氨基酸如5至19个氨基酸的内部缺失，其中所述内部缺失在与SEQIDNO:1的氨基酸116至134对应的区域内，其中所述内部缺失由长度为0至12个氨基酸的置换肽置换；和ii9个或少于9个额外氨基酸取代、缺失和或插入。本文还提供包含本文所述的修饰的FGF-21多肽的任一种和药学上可接受的载剂或赋形剂的组合物。本文提供修饰的FGF-21多肽，其包含具有选自SEQIDNO:1-7的氨基酸序列的多肽，只是其中所述氨基酸序列包含：i2至19个氨基酸如5至19个氨基酸的内部缺失，其中所述内部缺失在与SEQIDNO:1的氨基酸116至134对应的区域内，其中所述内部缺失由长度为0至12个氨基酸的置换肽置换；和ii9个或少于9个额外氨基酸取代、缺失和或插入。本文还提供在对其有需要的患者中调节葡萄糖及脂质稳态、葡萄糖摄入、GLUT1表达和或葡萄糖、甘油三酯、胰岛素或胰高血糖素血清浓度的至少一项的方法，其包括向患者施用治疗有效量的本文所述的修饰的FGF-21多肽。本文还提供在对其有需要的患者中或所述患者的血液、血清样品或另外的样品中增加胰岛素敏感性、增加脂联素水平、降低血糖水平、降低胰高血糖素水平、降低甘油三酯水平、降低果糖胺水平、降低低密度胆固醇水平或降低C反应性蛋白水平的方法，其包括向患者施用治疗有效量的本文所述的修饰的FGF-21多肽。本文还提供在对其有需要的患者中治疗选自如下的病况或病症的方法：肥胖、糖尿病、胰腺炎、胰岛素抗性、高胰岛素血症、葡萄糖不耐症、高血糖症、代谢综合征、葡萄糖耐量受损、葡萄糖清除不足、高血糖、A型胰岛素抗性、C型胰岛素抗性AKAHAIR-AN综合征、罗伯森-门登荷综合征Rabson-MendenhallSyndrome、多诺霍氏综合征Donohue'sSyndrome或矮怪病Leprechaunism、雄激素过多症、多毛症或黑棘皮病、和普拉德-威利综合征Prader-Willisyndrome，其包括向患者施用治疗有效量的本文所述的修饰的FGF-21多肽。本文还提供在对其有需要的患者中治疗1型糖尿病、2型糖尿病或肥胖的方法，其包括向患者施用治疗有效量的本文所述的修饰的FGF-21多肽。本文还提供治疗纤维化相关疾病的方法，其包括向对其有需要的患者施用有效量的本文所述的修饰的FGF-21多肽。本文还提供治疗纤维化相关疾病的方法，其包括向对其有需要的患者施用有效量的包含本文所述的修饰的FGF-21多肽和药学上可接受的载剂或赋形剂的组合物。本文还提供在对其有需要的患者中治疗肝纤维化或肝硬化的方法，其包括向患者施用治疗有效量的本文所述的修饰的FGF-21多肽。本文还提供在对其有需要的患者中治疗或预防NASH的方法，其包括向患者施用治疗有效量的本文所述的修饰的FGF-21多肽。本文还提供治疗NASH和或肝纤维化的方法，其包括向对其有需要的患者施用有效量的包含本文所述的修饰的FGF-21多肽和药学上可接受的载剂或赋形剂的组合物。本文还提供在对其有需要的患者中降低肝脂肪分数的方法，其包括向患者施用治疗有效量的本文所述的修饰的FGF-21多肽，其中所述患者任选地处于患上NASH的风险或已诊断患有NASH。本文还提供在对其有需要的患者中增加脂联素水平如血浆总脂联素和或高分子量脂联素的方法，其包括向患者施用治疗有效量的本文所述的修饰的FGF-21多肽，其中所述患者任选地处于患上NASH的风险或已诊断患有NASH。本文还提供在对其有需要的患者中治疗心脏衰竭或心脏纤维化的方法，其包括向患者施用治疗有效量的本文所述的修饰的FGF-21多肽。本文还提供在对其有需要的患者中治疗肾脏或肾纤维化的方法，其包括向患者施用治疗有效量的本文所述的修饰的FGF-21多肽。本文还提供在对其有需要的患者中治疗肺纤维化的方法，其包括向患者施用治疗有效量的本文所述的修饰的FGF-21多肽。本文还提供在对其有需要的患者中治疗纤维化相关疾病的方法，其包括向患者施用有效量的包含一个或多个非天然编码的氨基酸的修饰的FGF-21多肽，其中所述修饰的FGF-21多肽与具有选自SEQIDNO:1-7和201的氨基酸序列的人FGF-21多肽具有至少90％同一性，其中所述纤维化相关疾病选自NASH、肝纤维化、糖尿病性肾脏疾病、慢性肾病、肾纤维化、肺纤维化、心脏纤维化、心脏衰竭及代谢性心脏衰竭。附图简述图1A至B.例示性的修饰的FGF-21多肽序列。指示的坐标相对于SEQIDNO:1的野生型FGF-21多肽。完整多肽各包含N末端序列MHPIPDSSPLLQFGGQVRQRYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRELLLEDGYNVY，然后是位置108处的指示氨基酸，然后是标记为“氨基酸109-149”的区域的指示序列，然后是序列PGILAPQPPDVGSSDPLSM，然后是标记为“氨基酸169-181”的区域的指示序列。修饰的FGF-21多肽的完整氨基酸序列还示于下文实施例4中。图2.聚乙二醇化的化合物1及聚乙二醇化的化合物2的体外FGF-21活性的例示性剂量反应曲线在基于细胞的测定法中通过胞外信号调节激酶ERK12的FGF21依赖性磷酸化测量。图3.用于评估修饰的FGF-21多肽的热稳定性进行的差示扫描量热法DSC的代表性结果。在此实例中，显示了聚乙二醇化的化合物2具有高出聚乙二醇化的化合物1约8℃的转变中点“Tm”温度。热可逆性95％未显示。图4.用于评估修饰的FGF-21多肽的热稳定性进行的热扫描荧光分析TSF的代表性结果。在此图中，显示了化合物10具有高出化合物1具有野生型FGF-21序列，但其中包括N末端甲硫氨酸以在大肠杆菌E.coli中表达约8℃的转变中点“Tm”温度。图5.左小图：25℃聚乙二醇化的化合物1及聚乙二醇化的化合物2随时间的脱酰胺化的测量通过电荷异质性表示。聚乙二醇化的化合物2表现出减少的电荷异质性形成，指示脱酰胺化大幅减慢。右小图：40℃各自处于pH7.0和7.5mgml的蛋白质浓度的蔗糖组氨酸缓冲液中的聚乙二醇化的化合物1及聚乙二醇化的化合物2随时间的聚集体形成的测量通过大小排阻色谱测量。聚乙二醇化的化合物2表现出减少的聚集体形成。总之，这些结果能够预测相对于聚乙二醇化的化合物1，聚乙二醇化的化合物2具有更佳的储存稳定性和配制至较高浓度的能力。图6.在pH6.5处于蔗糖组氨酸缓冲液中上方小图、在pH7.0处于蔗糖组氨酸缓冲液中中间小图及在pH8.3处于蔗糖TRIS缓冲液中下方小图修饰的FGF-21多肽随时间的高分子量HMW聚集体形成的测量。相比于pH8.3处，在pH6.5及pH7.0处的HMW聚集体形成更为明显。大多数FGF-21变体表现出相对于聚乙二醇化的化合物1减少的HMW聚集体形成，但对于少数化合物，HMW聚集体形成与化合物1类似或更高。图7.在pH6.5处于蔗糖组氨酸缓冲液中上方小图、在pH7.0于蔗糖组氨酸缓冲液中中间小图及在pH8.3于蔗糖TRIS缓冲液中下方小图修饰的FGF-21多肽的脱酰胺化倾向的测量通过随时间的酸性变体的形成表示。与聚乙二醇化的化合物1相比，所有所示化合物都表现出减少的电荷酸性变体形成，指示降低的脱酰胺化。图8.修饰的FGF-21多肽的溶解度的测量。给定多肽浓度处相对较低的高分子量聚集体形成指示较大溶解度，由此使得能够配制为较大浓度。测试化合物的蛋白质浓度相对于高分子量HMW种类的百分比的观察结果经由pH7.2处的PBS缓冲液中的活塞流过滤测定法plugflowfiltrationassay来确定。线性拟合至这些观察结果中的每一项的线性斜率用于估计蛋白质聚集倾向。图9.在7天蛋白和外周血液单核细胞PBMC温育之后具有CD4+T细胞增殖响应的供体的百分比。对照蛋白是具有已知T细胞活化特性的大肠杆菌表达的蛋白。比较非聚乙二醇化经修饰的FGF-21化合物与化合物1具有野生型FGF-21序列，但其中包括N末端甲硫氨酸以在大肠杆菌中表达。“ConA”是伴刀豆球蛋白A一种已知免疫原性蛋白的缩写。图10.在向obob小鼠单次S.C.施用0.05mgkg之后聚乙二醇化的化合物1及聚乙二醇化的化合物2的药代动力学分析。修饰的FGF-21化合物的水平以总计与C-末端完整活性多肽二者测量。与聚乙二醇化的化合物1相比，聚乙二醇化的化合物2表现出对于C-末端完整形式大得多的总AUC，指示聚乙二醇化的化合物2的体内蛋白水解大幅降低。结果图示于上方小图中且药代动力学参数列表于左下方小图聚乙二醇化的化合物1及右下方小图聚乙二醇化的化合物2中。图11A至B.向食蟹猴cynomolgusmonkeys施用后的聚乙二醇化的化合物1和聚乙二醇化的化合物2的药代动力学分析。修饰的FGF-21化合物的水平以A总计和BC-末端完整活性多肽测量。与聚乙二醇化的化合物1相比，聚乙二醇化的化合物2表现出对于C-末端完整形式大得多的总AUC，指示聚乙二醇化的化合物2的体内蛋白水解大幅降低。图12.obob小鼠中重复给药研究的第21天的相对于介质的糖化血红蛋白HbA1c变化。图13.obob小鼠中重复给药研究的21天的体重。图14.obob小鼠中重复给药研究的21天的体重百分比变化。图15.obob小鼠中21天重复给药研究期间的血浆胰岛素浓度。图16.obob小鼠中21天重复给药研究期间的血浆葡萄糖浓度。图17.obob小鼠中21天重复给药研究期间的AUC血浆葡萄糖浓度变化，表示为与介质处理对照的百分比差异。图18.obob小鼠中21天重复给药研究期间的血浆甘油三酯浓度。图19.StelicNASH小鼠研究中的体重变化。图20.StelicNASH小鼠研究中的总食品消耗。图21.StelicNASH小鼠研究中的体重。图22.StelicNASH小鼠研究中的肝脏重量。图23.StelicNASH小鼠研究中的肝-体重比。图24.StelicNASH小鼠研究中的全血葡萄糖。图25.StelicNASH小鼠研究中的血浆ALT。图26.StelicNASH小鼠研究中的血浆甘油三酯。图27.StelicNASH小鼠研究中的总血浆胆固醇。图28.StelicNASH小鼠研究中的肝脏甘油三酯。图29.StelicNASH小鼠研究中的肝脏胆固醇。图30A至B.StelicNASH小鼠研究中HE染色肝脏切片的代表性显微照片。图31.StelicNASH小鼠研究中的NAFLD活性评分。图32A.StelicNASH小鼠研究中的脂肪变性评分。图32B.StelicNASH小鼠研究中的小叶炎症评分。图32C.StelicNASH小鼠研究中的肝细胞鼓起评分。图33A至B.StelicNASH小鼠研究中的天狼星红染色肝脏切片的代表性显微照片。图34.StelicNASH小鼠研究中的纤维化区域。图35A至B.StelicNASH小鼠研究中的F480免疫染色肝脏切片的代表性显微照片。图36.StelicNASH小鼠研究中的炎症区域。图37A至B.StelicNASH小鼠研究中油红染色肝脏切片的代表性显微照片。图38.StelicNASH小鼠研究中的脂肪沉积区域。图39A至D.StelicNASH小鼠研究中的相对基因表达。Aα-SMA、BTIMP-1、C1型胶原和DTGF-β的表达结果。图40A至D.融合至Fc域、PKEAdnectin或人血清白蛋白多肽的例示性修饰的FGF-21多肽序列。“融合排列”描述了融合蛋白的由N至C末端的通常取向，其意在说明但不限于对应多肽序列。HuSAC34A：具有氨基酸序列SEQIDNO:321的修饰的人血清白蛋白；HuSAC34A；desLeu-585：具有氨基酸序列SEQIDNO:322的修饰的人血清白蛋白；PKE1：具有氨基酸序列SEQIDNO:319的PKEAdnectin；PKE2：具有氨基酸序列SEQIDNO:320的PKEAdnectin。后面有数字的L表示接头；FGF-21Cmp.2表示含有内部缺失及置换肽的FGF-21序列具体而言为化合物2且FGF-21Cmp.1表示缺乏所述缺失及置换肽的FGF-21序列具体而言为化合物1。FGF-21Cmp.1和FGF-21Cmp.2可包括或缺乏化合物1及2中所含有的N末端甲硫氨酸。Fc多肽序列的变体形式以圆括号标识，例如FchIgG1a_191。标记FGF-21-1aaCmp.2和FGF-21-3aaCmp.2是指通过自其C末端缺失所述数量的氨基酸分别为1或3而修饰的化合物2序列。图41.融合至Adnectin融合配偶体的修饰的FGF-21多肽的溶解度的测量。给定多肽浓度处相对较少的高分子量聚集体形成指示较大溶解度，由此使得能够配制为较大浓度。测试化合物的蛋白质浓度相对于高分子量HMW种类的百分比的观察结果经由pH为7.2的PBS缓冲液中的活塞流过滤测定法来确定。线性拟合至这些观察结果中的每一项的线性斜率用于估计蛋白质聚集倾向。图42A至B.StelicNASH小鼠研究中处理组的体重。A.用介质或PEG-化合物“PEG-Cmpd”1处理9至12周的小鼠的比较。B.用介质或PEG-化合物1处理9-15周的小鼠的比较。图43A至B.StelicNASH小鼠研究中处理组的肝脏重量。A.用介质或PEG-化合物1处理9-12周的小鼠的比较。B.用介质或PEG-化合物1处理9-15周的小鼠的比较。经9-15周处理的小鼠的肝脏重量显著降低p0.5。图64.基于细胞的Elk1-荧光素酶测定法中测量的FGF-21变体的效力。各数据点指示个体重复且水平棒表示均值。值表示为以摩尔计的测量的EC50值的以10为底的对数。图65.C57BL6J小鼠中单次急性剂量的PEG-化合物2对百分比体重变化的影响。用介质250mM蔗糖20mMTris，pH8.3或0.03、0.1、0.3或1.0mgkg的PEG-化合物2处理C57BL6J小鼠，n＝10组。确定基线0、SC给药后24、48、72和144h时C57BL6J小鼠的百分比体重变化。所有值为均值±SEM*P95％数据未显示。图4显示了TSF的代表性结果。化合物10表现出相对于化合物1具有野生型FGF-21序列，但其包括N末端甲硫氨酸以在大肠杆菌中表达Tm增加约8℃。其他化合物的结果示于下表6中。总之，这些结果说明可合理预期例示性化合物会降低形成聚集体的倾向，例如化合物2、10和16。预期降低的形成聚集体的倾向会赋予更长的储存期限和或配制为较大浓度的能力。表6.通过热扫描荧光确定修饰的FGF-21多肽相比于化合物1的热稳定性结果。对于一些化合物观察到了多至约7-8℃的热稳定性增加。实施例7脱酰胺化和聚集体形成本实施例描述了对修饰的FGF-21多肽的脱酰胺化和聚集体形成的评估。方法热应力测试：在测试缓冲液250mM蔗糖，20mM组氨酸，pH6.5和pH7或20mMTRISpH7.5中以7.5mg蛋白mL的浓度制备目的蛋白样品。通过分析型大小排阻色谱aSEC和时间＝0时的成像毛细管等电聚焦icIEF的电荷变体分析来测验样品。然后通过将样品置于25℃培养箱中1周然后在40℃5周来测试样品的应力。取出等分试样并通过icIEF和aSEC在各个时间点进行检查，在接下来的4至5周取出并进一步通过aSEC和icIEF进行检查以监测稳定性。aSEC使用Agilent1100HPLC系统AgilentTechnologies,SantaClara,CAUSA在Zenix-C300SEC柱SepaxTechnologies,Newark,DEUSA上用300×4.6mm的尺寸进行大小排阻色谱aSEC。所用的流动相是200mM磷酸钾和150mM氯化钠，其调节至pH6.9，流动速率为0.35mLmin。对于各分析注射约20μg样品。在UV280nm处监测分离。在与各物质对应的保留时间处通过曲线下面积的积分来进行单体对HMW物种的定量。icIEF通过成像毛细管等电点聚焦icIEF进行的电荷变体分析脱酰胺化会增加蛋白的净负电荷和酸性变体的形成检测脱酰胺化，其在iCE280仪器ProteinSimple,SantaClara,CAUSA上使用经碳氟化合物涂布的毛细管100μm×50mm进行。使用含有0.35％甲基纤维素、4％pH3-10Pharmalytes、4M尿素和2％pI标志物的溶液将样品稀释至0.3mgmL。阳极电解液和阴极电解液分别为0.08M磷酸和0.1M氢氧化钠。通过应用1.5kV2min然后3.0kV11min来达到聚焦。由对应的曲线下面积的积分，相对于亲本物质，来定量-电荷的修饰的物种。结果聚乙二醇化的化合物2的生物物理学特性表现出相对于聚乙二醇化的化合物1的优越性。在2-8℃经5周均未观察到降解，伴随着在40℃在加速条件下观察到相对于聚乙二醇化的化合物1的低速降解图5。八周加速研究指示，在测试配制物组氨酸蔗糖pH7.0中聚乙二醇化的化合物2优于聚乙二醇化的化合物1。在这些赋形剂条件下，脱酰胺化消除且聚乙二醇化的化合物2降低了形成可溶性高分子量HMW聚集体的倾向。这些结果提示，相对于聚乙二醇化的化合物1，聚乙二醇化的化合物2具有降低的聚集倾向，具有可供配制选择的较宽pH范围。还分析了HMW聚集体形成。在给定浓度处HMW聚集体形成较少表示溶解度较大，由此使得能够配制为较高浓度。图6说明了数种修饰的FGF-21多肽和化合物1的HMW聚集体形成。对于大多数测试化合物，相对于聚乙二醇化的化合物1的聚集体形成降低例如聚乙二醇化的化合物2和聚乙二醇化的化合物10。对于少数化合物，HMW聚集体形成类似于或高于聚乙二醇化的化合物1。还分析了脱酰胺化。图7说明数种修饰的FGF-21多肽和聚乙二醇化的化合物1所检测的脱酰胺化水平由随时间的酸性变体的形成指示。对于所有显示的修饰的FGF-21多肽，检测到的脱酰胺化水平是相对于聚乙二醇化的化合物1降低的。实施例8修饰的FGF-21多肽的溶解度评估本实施例描述了修饰的FGF-21多肽的相对溶解度的测量。结果显示化合物能够配制成相对高的浓度，其允许更便利地施用有效剂量。方法相对溶解度评估通过序贯活塞流浓缩循环，然后是大小排阻色谱分析来进行。将于PBSpH7.2中以类似但不相同的起始浓度配制的样品吸入至3kDa分子量截留离心浓缩器中并在4℃以4,750RPM旋转10分钟、15分钟和30分钟的循环。在旋转循环之间，自浓缩装置移出等分试样并进行分析型大小排阻色谱分析aSEC在用PBSpH7.2缓冲液平衡的GEHealthcareSuperdexS-7510300GL柱上以确定溶液中HMW及单体物种的浓度。用NanoDrop分光光度计通过280nm处的吸光度来确定总浓度。相对于SEC色谱图迹线中的单体峰，通过高分子量峰的曲线下面积计算来确定高分子量百分比。绘制所得数据点以将观察测试条件下从最难溶构建体至最可溶的排列顺序可视化，由数据点产生的斜率越低，测试条件下的蛋白变体越稳定。结果修饰的FGF-21多肽的相对溶解度通过测量高分子量HMW聚集体的形成作为蛋白浓度的函数来确定。在给定浓度下较低的HMW聚集体形成指示较大的溶解度，由此使得能够配制至较高浓度。测试化合物的活塞流溶解度曲线的斜率示于下表7中较低值表示较少聚集体形成且因此能够配制为较高浓度。对于其中数种化合物例如化合物2和化合物10观察到相对低的聚集体形成水平，指示较低的聚集体形成和较大的溶解度图8和表7。观察到化合物1的活塞流溶解度斜率最高。表7.修饰的FGF-21多肽的活塞流溶解度结果化合物活塞流溶解度斜率化合物11.863化合物20.3171化合物31.2219化合物50.8548化合物61.1353化合物71.4318化合物100.3647化合物111.1156化合物121.0317化合物190.9743化合物201.1545化合物220.1997化合物230.771实施例9FGF-21缺失分子的免疫原性测试进行T细胞活化响应研究，并指示大多数化合物中所用的蛋白序列与聚乙二醇化的化合物1中所用的蛋白序列表现出等同响应。方法通过CD4+T细胞增殖测定评估免疫原性。使用Ficoll梯度自不同组的人血液供体分离外周血单核细胞PBMC。对供体进行HLA分型以确保覆盖人群体中存在的II类MHC等位基因的变化。在标记之后，具有羧基荧光素琥珀酰亚胺酯CFSE的PBMC以每孔200,000个细胞在测试化合物存在下于标准细胞培养基中平板接种于96孔格式中7天。通过用抗CD4抗体标记和流式细胞术分析CD4+T细胞的增殖。抗原性蛋白的百分比计算为相对于介质对照表现出显著CD4+T细胞增殖响应的供体的百分比。结果在利用人T细胞增殖测定的体外免疫原性风险评估中，与40个供体中的15个显示出来自化合物1具有野生型FGF-21序列，但其包括N末端甲硫氨酸以在大肠杆菌中表达阳性信号相比，在暴露于化合物2经7天之后，40个供体中的13个32.5％显示出CD4+增殖响应图9。尽管此基于细胞的实验不能替代实际世界的人免疫响应观察结果，但此分析指示与野生型FGF21相比，化合物2的人免疫原性风险没有增加。在相对于化合物1的CD4+增殖测定中测试了进一步修饰的FGF-21多肽图9。免疫原性一般与对照FGF21序列化合物1观察到的免疫原性类似，但其中例外为化合物10，其表现出相对较更高的免疫响应，指示若向人患者施用，则该化合物可能具有不合意的免疫原性。实施例10C末端完整活性的修饰的FGF-21多肽的体内稳定性本研究评估了体内C末端完整即活性的修饰的FGF-21多肽的水平。与聚乙二醇化的化合物1相比，聚乙二醇化的化合物2表现出大大增加的C末端完整的活性多肽的比例，包括更长的体内活性的持续时间。方法在食蟹猴中在分别皮下SC给药0.25mgKg和0.225mgkg之后评估聚乙二醇化的化合物2和聚乙二醇化的化合物1的药代动力学。在药物施用之后0、0.25、0.5、1、3、7、24、48、72、96、120、144、168小时收集血液样品0.2mL并储存于-80℃直至进行生物分析。总计和C末端完整的聚乙二醇化的化合物1及聚乙二醇化的化合物2的PK参数通过血清或血浆浓度相对于时间数据的非分区分析non-compartmentalanalysis获得PhoenixTM6.3,PharsightCorporation,St.Louis,MO。使用线性和对数梯形求和的组合计算时间零点至无穷大[AUCtot]的曲线下面积。可以定量浓度使用3个时间点的最小值估计AUC和半衰期t12。使用未验证的基于中型探索MSD的电化学发光免疫吸附剂分析ECLIA测量了单次剂量PK研究中总聚乙二醇化的化合物1的浓度。使用PEG特异性单克隆抗体mAb捕获聚乙二醇化的化合物1，随后使用兔抗FGF21多克隆抗体pAb检测总聚乙二醇化的化合物1。还在雄性ObOb小鼠中评估了聚乙二醇化的化合物2和聚乙二醇化的化合物1的皮下SC药代动力学PK。向小鼠n＝3时间点途径以单次皮下0.1mgkg注射施用聚乙二醇化的化合物2。施用0.05mgkg呈单次皮下剂量形式的聚乙二醇化的化合物1。在药物施用后的各个时间点收集血液样品并基本上如对于食蟹猴所述进行处理。经由电化学发光读出，以相对光度单位RLU表示，其与通过捕获和检测试剂结合的总聚乙二醇化的化合物1的量成比例。标准曲线在0.2与154ngmL之间。使用4参数对数logistic拟合回归模型用1Y的权重因子定量测试样品。使用相同的测定来测量ZDF大鼠和食蟹猴血清中C末端完整的聚乙二醇化的化合物1的浓度，只是使用特异性针对聚乙二醇化的化合物1的C末端的兔抗FGF21pAb进行检测。标准曲线在0.2与154ngmL之间。使用4参数对数拟合回归模型用1Y的加权因子定量测试样品。结果聚乙二醇化的化合物1经历体内蛋白水解，以使得随时间大多数化合物变为截短的非活化形式。在obob小鼠图10和食蟹猴图11中直接比较聚乙二醇化的化合物1和2的体内蛋白水解稳定性。这些结果显示了与聚乙二醇化的化合物1相比，含有缺失的修饰的FGF-21化合物聚乙二醇化的化合物2的活性C末端完整形式的AUC增加7至8倍，指示聚乙二醇化的化合物2的体内蛋白水解稳定性增加。实施例11修饰的FGF-21多肽的药代动力学研究本实施例显示了在在小鼠、大鼠及食蟹猴中使用修饰的FGF-21多肽进行的药代动力学研究的结果参见下表8。简言之，在小鼠、大鼠和猴中聚乙二醇化的化合物2的清除率低范围：0.94-2.6mLhkg。约0.04-0.05Lkg的分布体积Vss接近血浆体积且指示血管外间隙中的最少分布。在小鼠40％和猴58％中的皮下SC生物利用度是可接受的，而在大鼠16％中其相对低。此低皮下生物利用度可为特异性针对大鼠的，因为已观察到其他聚乙二醇化分子具有类似现象在大鼠中具有低SC生物利用度，在小鼠、猴和人中具有良好生物利用度。更重要的是，这些物种包括大鼠中聚乙二醇化的化合物2的SC生物利用度比聚乙二醇化的化合物1高2-3.4倍。在向小鼠、大鼠及猴皮下施用聚乙二醇化的化合物2之后，相对于聚乙二醇化的化合物1，完整蛋白的剂量校正的暴露AUC一致7-8倍增加。在所有三种物种中，相对于聚乙二醇化的化合物1，在皮下施用之后完整聚乙二醇化的化合物2的暴露增加可归因于较低的全身清除率2-5倍以及皮下生物利用度改进2-3.4倍。由蛋白水解片段和完整聚乙二醇化的化合物2的混合物构成的总聚乙二醇化的化合物2也以这些药代动力学研究测量。在小鼠和大鼠中，在静脉内施用之后完整的聚乙二醇化的化合物2相对于总聚乙二醇化的化合物2的AUC比率接近1，从而指示系统性蛋白水解最少。然而，在这些物种中，在皮下施用之后该比率为0.6-0.86，指示了在SC部位的一些蛋白水解。在猴中注意到了类似趋势，其中与静脉内施用相比，在SC施用之后完整聚乙二醇化的化合物2相对于总聚乙二醇化的化合物2的AUC比率有少量降低在SC给药之后为0.7对比在IV给药之后为0.8。总体而言，聚乙二醇化的化合物2表现出有利的体外及体内药代动力学特性，包括增加的生物利用度、降低的清除率及增加的AUC血浆浓度-时间曲线下面积。表8.小鼠、大鼠和猴中完整的修饰的FGF-21蛋白的IV和SC药代动力学缩写：CL：血浆的表观总身体药物清除率；Vss：稳定状态下的表观分布体积；MRT：平均保留时间；SCBA：皮下生物利用度；SCAUC：皮下血浆浓度-时间曲线下面积。实施例12糖尿病小鼠模型中修饰的FGF-21多肽的体内重复给药研究在21天重复给药研究中直接head-to-head评估聚乙二醇化的化合物2和聚乙二醇化的化合物1。结果指示尽管两种化合物对于改善代谢症状都有效，但较长的活性聚乙二醇化的化合物2的体内半衰期会产生更大的治疗作用，在比较每周给药的影响时尤其如此。方法在本研究开始时雄性obob小鼠JacksonLaboratories,BarHarbor,ME为8周龄。基于体重和葡萄糖水平将小鼠随机分入处理组。所有组通过皮下s.c.施用1mlkg给药溶液处理。处理组如下：1介质250mM蔗糖20mMTris，pH8.3，每周两次BIW，2聚乙二醇化的化合物1，0.15mgkgBIW，3聚乙二醇化的化合物2，0.15mgkgBIW，4聚乙二醇化的化合物1，0.3mgkg，每周一次QW，或5聚乙二醇化的化合物2，0.3mgkgQW。每周仅施用一次化合物的小鼠在BIW组施用其每周第二次化合物注射剂之日时施用介质。确定饲喂状态下整个21天给药期中的体重、血浆葡萄糖、甘油三酯Olympus临床化学分析仪，AU680和胰岛素ELISA，MercodiaInc.。在研究开始和终止时也用Olympus分析仪确定全血中的糖化血红蛋白HbA1c。结果皮下施用0.15mgkg的剂量每周两次BIW和0.3mgkgQW施用聚乙二醇化的化合物1与聚乙二醇化的化合物2。第21天在最低处测量的血浆浓度显示总完整和蛋白水解的FGF21的暴露对于两种变体相似，但在QW或BIW施用之后，活性C末端完整聚乙二醇化的化合物2的暴露分别比聚乙二醇化的化合物1高12至25倍表9。表9：在obob小鼠中在重复给药21天之后最低处的C末端完整的聚乙二醇化的化合物1和聚乙二醇化的化合物2的暴露平均值±s.e.m.*基于obob小鼠中的单剂量药代动力学数据预测从将葡萄糖无酶催化添加至此蛋白内的特异性氨基酸中体内产生糖化血红蛋白HbA1c，且测量的HbA1c百分比与循环红细胞的寿命上并入的平均血糖对应。大于6.5％的HbA1c值是糖尿病的诊断准则。图12中显示出重复给药研究的第21天的介质校正的HbA1c变化。BIW施用0.15mgkg的任一FGF21多肽在施用21天之后将HbA1c校正至小于5％的值。然而，与以QW施用0.3mgkg介质相比，仅聚乙二醇化的化合物2在统计上显著降低ΔHbA1c，这与对于活性C末端完整蛋白的功能性暴露显著增加是一致的。这些结果表面维持目标最低浓度C最低处对于功效可为足够的表9。与降低的HbA1c结果一致，在整个研究中血浆葡萄糖水平也显著降低第2至21天，相对于介质的所有组p0.05。在处理四周之后在研究结束时，PEG-化合物2组中的血浆葡萄糖低于介质组23％485±39对比629±43,Pb628.6±39.0bUnixdbdb,PEG-化合物2628.6±39.0b485.2±42.5ca.数据表示为均值±S.E.M。b.P30μgmg是糖尿病性患者中最敏感性和特异性的早期肾病指标之一参见Chae等，JKoreanMedSci.2012；27:784-787，其通过提述以其整体并入本文。自3小时点尿收集物测量微量白蛋白并将值校正至尿液肌酐。在研究过程期间非糖尿病dbm瘦小鼠的平均uACR值范围为21至37μgmg图68和表28。在研究期间，与非糖尿病性瘦dbm小鼠相比，unixdbdb小鼠中的uACR由23显著增加至62倍所有Pb1060.2±256.9b1319.9±282.3b1341.3±283.5bUnixdbdb,PEG-化合物2865.8±170.8538.5±107.5c616.1±145.2c460.6±92.7c肾脏湿重反映了由于糖尿病中的超滤的肾肥大的状态。与非糖尿病性瘦小鼠相比，在unixdbdb小鼠中平均肾脏重量显著较大，用介质处理的unixdbdb小鼠的肾脏重量增加59％。用PEG-化合物2处理四周显著抑制糖尿病小鼠中的肾脏重量增加286±6.3对比253±7.4，介质对比PEG-化合物2，Pa98911162523335-肾小管间质b3341224211-骨盆c21212-12-11a肾小球膜扩张是局部或弥散性的，且部分或总体的。2级是基于50％的具有膜扩张的肾小球评估和或至少1个肾小球中的额外肾小球膜、毛细管、上皮或荚膜病损；3级是基于这些额外肾小球病损的多于一项。b小管病损包括膨胀、萎缩和或增生；间质病损通常与肾小球病损相关且其特征为最小炎症或局部管周纤维化。1级为局部；2级为具有2至4个病灶的多灶性的。c化脓性炎症；1级基于限于骨盆上皮的炎症；2级基于延伸至髓质或骨盆周围皮层。实施例20.对异丙肾上腺素灌注的BalbC小鼠中的心脏肥大和心脏纤维化的影响异丙肾上腺素Iso灌注的Balbc小鼠是心脏纤维化和肥大的啮齿动物模型。将10至12周的雄性Balbc小鼠基于药物施用前的体重随机化。经由渗透微泵皮下施用Iso或介质于盐水中的0.02％抗坏血酸3周。小鼠经3周的过程每周两次BIW同时给药0.5mgkgPEG化合物2或培养基。然后使动物安乐死并在大血管处移出心脏并轻吸以去除心室中的任何血液，然后称重。自各组中的剩余24只小鼠，将整个心脏于干冰上冷冻并储存于≤-70℃以进行羟脯氨酸HP分析。使用总胶原测定试剂盒根据制造商的说明书QuickzymeBiosciences,Netherlands进行心脏HP含量心脏纤维化终点分析。在3周生命期结束时，测量整个心脏HP含量。与假拟物+介质组9.96±0.38μgmg相比，在Iso+介质组21.35±0.81μgmg中，经校正的HP含量增加114％P百时美施贵宝公司修饰的FGF-21多肽及其用途43270.22132015-10-23US62068,2962014-10-24US62068,5142014-10-24US62068,5232014-10-24US62068,5262014-10-24US62068,5342014-10-24US62141,3372015-04-01US62141,3832015-04-01487PatentInversion3.51181PRT人Homosapiens1HisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGlnVal151015ArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAlaHis202530LeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGlnSer354045ProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIleGln505560IleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAspGly65707580AlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPheArg859095GluLeuLeuLeuGluAspGlyTyrAsnValTyrGlnSerGluAlaHis100105110GlyLeuProLeuHisLeuProGlyAsnLysSerProHisArgAspPro115120125AlaProArgGlyProAlaArgPheLeuProLeuProGlyLeuProPro130135140AlaProProGluProProGlyIleLeuAlaProGlnProProAspVal145150155160GlySerSerAspProLeuSerMetValGlyProSerGlnGlyArgSer165170175ProSerTyrAlaSer1802191PRT人Homosapiens2MetHisHisHisHisHisHisSerGlyGlyHisProIleProAspSer151015SerProLeuLeuGlnPheGlyGlyGlnValArgGlnArgTyrLeuTyr202530ThrAspAspAlaGlnGlnThrGluAlaHisLeuGluIleArgGluAsp354045GlyThrValGlyGlyAlaAlaAspGlnSerProGluSerLeuLeuGln505560LeuLysAlaLeuLysProGlyValIleGlnIleLeuGlyValLysThr65707580SerArgPheLeuCysGlnArgProAspGlyAlaLeuTyrGlySerLeu859095HisPheAspProGluAlaCysSerPheArgGluLeuLeuLeuGluAsp100105110GlyTyrAsnValTyrGlnSerGluAlaHisGlyLeuProLeuHisLeu115120125ProGlyAsnLysSerProHisArgAspProAlaProArgGlyProAla130135140ArgPheLeuProLeuProGlyLeuProProAlaProProGluProPro145150155160GlyIleLeuAlaProGlnProProAspValGlySerSerAspProLeu165170175SerMetValGlyProSerGlnGlyArgSerProSerTyrAlaSer1801851903209PRT人Homosapiens3MetAspSerAspGluThrGlyPheGluHisSerGlyLeuTrpValSer151015ValLeuAlaGlyLeuLeuLeuGlyAlaCysGlnAlaHisProIlePro202530AspSerSerProLeuLeuGlnPheGlyGlyGlnValArgGlnArgTyr354045LeuTyrThrAspAspAlaGlnGlnThrGluAlaHisLeuGluIleArg505560GluAspGlyThrValGlyGlyAlaAlaAspGlnSerProGluSerLeu65707580LeuGlnLeuLysAlaLeuLysProGlyValIleGlnIleLeuGlyVal859095LysThrSerArgPheLeuCysGlnArgProAspGlyAlaLeuTyrGly100105110SerLeuHisPheAspProGluAlaCysSerPheArgGluLeuLeuLeu115120125GluAspGlyTyrAsnValTyrGlnSerGluAlaHisGlyLeuProLeu130135140HisLeuProGlyAsnLysSerProHisArgAspProAlaProArgGly145150155160ProAlaArgPheLeuProLeuProGlyLeuProProAlaProProGlu165170175ProProGlyIleLeuAlaProGlnProProAspValGlySerSerAsp180185190ProLeuSerMetValGlyProSerGlnGlyArgSerProSerTyrAla195200205Ser4208PRT人Homosapiens4MetAspSerAspGluThrGlyPheGluHisSerGlyLeuTrpValSer151015ValLeuAlaGlyLeuLeuGlyAlaCysGlnAlaHisProIleProAsp202530SerSerProLeuLeuGlnPheGlyGlyGlnValArgGlnArgTyrLeu354045TyrThrAspAspAlaGlnGlnThrGluAlaHisLeuGluIleArgGlu505560AspGlyThrValGlyGlyAlaAlaAspGlnSerProGluSerLeuLeu65707580GlnLeuLysAlaLeuLysProGlyValIleGlnIleLeuGlyValLys859095ThrSerArgPheLeuCysGlnArgProAspGlyAlaLeuTyrGlySer100105110LeuHisPheAspProGluAlaCysSerPheArgGluLeuLeuLeuGlu115120125AspGlyTyrAsnValTyrGlnSerGluAlaHisGlyLeuProLeuHis130135140LeuProGlyAsnLysSerProHisArgAspProAlaProArgGlyPro145150155160AlaArgPheLeuProLeuProGlyLeuProProAlaProProGluPro165170175ProGlyIleLeuAlaProGlnProProAspValGlySerSerAspPro180185190LeuSerMetValGlyProSerGlnGlyArgSerProSerTyrAlaSer1952002055181PRT人Homosapiens5HisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGlnVal151015ArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAlaHis202530LeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGlnSer354045ProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIleGln505560IleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAspGly65707580AlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPheArg859095GluLeuLeuLeuGluAspGlyTyrAsnValTyrGlnSerGluAlaHis100105110GlyLeuProLeuHisLeuProGlyAsnLysSerProHisArgAspPro115120125AlaProArgGlyProAlaArgPheLeuProLeuProGlyLeuProPro130135140AlaLeuProGluProProGlyIleLeuAlaProGlnProProAspVal145150155160GlySerSerAspProLeuSerMetValGlyProSerGlnGlyArgSer165170175ProSerTyrAlaSer1806209PRT人Homosapiens6MetAspSerAspGluThrGlyPheGluHisSerGlyLeuTrpValSer151015ValLeuAlaGlyLeuLeuLeuGlyAlaCysGlnAlaHisProIlePro202530AspSerSerProLeuLeuGlnPheGlyGlyGlnValArgGlnArgTyr354045LeuTyrThrAspAspAlaGlnGlnThrGluAlaHisLeuGluIleArg505560GluAspGlyThrValGlyGlyAlaAlaAspGlnSerProGluSerLeu65707580LeuGlnLeuLysAlaLeuLysProGlyValIleGlnIleLeuGlyVal859095LysThrSerArgPheLeuCysGlnArgProAspGlyAlaLeuTyrGly100105110SerLeuHisPheAspProGluAlaCysSerPheArgGluLeuLeuLeu115120125GluAspGlyTyrAsnValTyrGlnSerGluAlaHisGlyLeuProLeu130135140HisLeuProGlyAsnLysSerProHisArgAspProAlaProArgGly145150155160ProAlaArgPheLeuProLeuProGlyLeuProProAlaLeuProGlu165170175ProProGlyIleLeuAlaProGlnProProAspValGlySerSerAsp180185190ProLeuSerMetValGlyProSerGlnGlyArgSerProSerTyrAla195200205Ser7208PRT人Homosapiens7MetAspSerAspGluThrGlyPheGluHisSerGlyLeuTrpValSer151015ValLeuAlaGlyLeuLeuGlyAlaCysGlnAlaHisProIleProAsp202530SerSerProLeuLeuGlnPheGlyGlyGlnValArgGlnArgTyrLeu354045TyrThrAspAspAlaGlnGlnThrGluAlaHisLeuGluIleArgGlu505560AspGlyThrValGlyGlyAlaAlaAspGlnSerProGluSerLeuLeu65707580GlnLeuLysAlaLeuLysProGlyValIleGlnIleLeuGlyValLys859095ThrSerArgPheLeuCysGlnArgProAspGlyAlaLeuTyrGlySer100105110LeuHisPheAspProGluAlaCysSerPheArgGluLeuLeuLeuGlu115120125AspGlyTyrAsnValTyrGlnSerGluAlaHisGlyLeuProLeuHis130135140LeuProGlyAsnLysSerProHisArgAspProAlaProArgGlyPro145150155160AlaArgPheLeuProLeuProGlyLeuProProAlaLeuProGluPro165170175ProGlyIleLeuAlaProGlnProProAspValGlySerSerAspPro180185190LeuSerMetValGlyProSerGlnGlyArgSerProSerTyrAlaSer1952002058546DNA人Homosapiens8caccccatccctgactccagtcctctcctgcaattcgggggccaagtccggcagcggtac60ctctacacagatgatgcccagcagacagaagcccacctggagatcagggaggatgggacg120gtggggggcgctgctgaccagagccccgaaagtctcctgcagctgaaagccttgaagccg180ggagttattcaaatcttgggagtcaagacatccaggttcctgtgccagcggccagatggg240gccctgtatggatcgctccactttgaccctgaggcctgcagcttccgggagctgcttctt300gaggacggatacaatgtttaccagtccgaagcccacggcctcccgctgcacctgccaggg360aacaagtccccacaccgggaccctgcaccccgaggaccagctcgcttcctgccactacca420ggcctgccccccgcacccccggagccacccggaatcctggccccccagccccccgatgtg480ggctcctcggaccctctgagcatggtgggaccttcccagggccgaagccccagctacgct540tcctga5469576DNA人Homosapiens9atgcatcatcatcatcatcatagcggcggccaccccatccctgactccagtcctctcctg60caattcgggggccaagtccggcagcggtacctctacacagatgatgcccagcagacagaa120gcccacctggagatcagggaggatgggacggtggggggcgctgctgaccagagccccgaa180agtctcctgcagctgaaagccttgaagccgggagttattcaaatcttgggagtcaagaca240tccaggttcctgtgccagcggccagatggggccctgtatggatcgctccactttgaccct300gaggcctgcagcttccgggagctgcttcttgaggacggatacaatgtttaccagtccgaa360gcccacggcctcccgctgcacctgccagggaacaagtccccacaccgggaccctgcaccc420cgaggaccagctcgcttcctgccactaccaggcctgccccccgcacccccggagccaccc480ggaatcctggccccccagccccccgatgtgggctcctcggaccctctgagcatggtggga540ccttcccagggccgaagccccagctacgcttcctga57610630DNA人Homosapiens10atggactcggacgagaccgggttcgagcactcaggactgtgggtttctgtgctggctggt60cttctgctgggagcctgccaggcacaccccatccctgactccagtcctctcctgcaattc120gggggccaagtccggcagcggtacctctacacagatgatgcccagcagacagaagcccac180ctggagatcagggaggatgggacggtggggggcgctgctgaccagagccccgaaagtctc240ctgcagctgaaagccttgaagccgggagttattcaaatcttgggagtcaagacatccagg300ttcctgtgccagcggccagatggggccctgtatggatcgctccactttgaccctgaggcc360tgcagcttccgggagctgcttcttgaggacggatacaatgtttaccagtccgaagcccac420ggcctcccgctgcacctgccagggaacaagtccccacaccgggaccctgcaccccgagga480ccagctcgcttcctgccactaccaggcctgccccccgcacccccggagccacccggaatc540ctggccccccagccccccgatgtgggctcctcggaccctctgagcatggtgggaccttcc600cagggccgaagccccagctacgcttcctga63011627DNA人Homosapiens11atggactcggacgagaccgggttcgagcactcaggactgtgggtttctgtgctggctggt60cttctgggagcctgccaggcacaccccatccctgactccagtcctctcctgcaattcggg120ggccaagtccggcagcggtacctctacacagatgatgcccagcagacagaagcccacctg180gagatcagggaggatgggacggtggggggcgctgctgaccagagccccgaaagtctcctg240cagctgaaagccttgaagccgggagttattcaaatcttgggagtcaagacatccaggttc300ctgtgccagcggccagatggggccctgtatggatcgctccactttgaccctgaggcctgc360agcttccgggagctgcttcttgaggacggatacaatgtttaccagtccgaagcccacggc420ctcccgctgcacctgccagggaacaagtccccacaccgggaccctgcaccccgaggacca480gctcgcttcctgccactaccaggcctgccccccgcacccccggagccacccggaatcctg540gccccccagccccccgatgtgggctcctcggaccctctgagcatggtgggaccttcccag600ggccgaagccccagctacgcttcctga62712545DNA人Homosapiens12caccccatccctgactccagtcctctcctgcaattcgggggccaagtccggcagcggtac60ctctacacagatgatgcccagcagacagaagcccacctggagatcagggaggatgggacg120gtggggggcgctgctgaccagagccccgaaagtctcctgcagctgaaagccttgaagccg180ggagttattcaaatcttgggagtcaagacatccaggttcctgtgccagcggccagatggg240gccctgtatggatcgctccactttgaccctgaggcctgcagcttccgggagctgcttctt300gaggacggatacaatgtttaccagtccgaagcccacggcctcccgctgcacctgccaggg360aacaagtccccacaccgggaccctgcaccccgaggaccagctcgcttcctgccactacca420ggcctgccccccgcactcccggagccacccggaatcctggccccccagccccccgatgtg480ggctcctcggaccctctgagcatggtgggaccttcccagggccgaagcccagctacgctt540cctga54513630DNA人Homosapiens13atggactcggacgagaccgggttcgagcactcaggactgtgggtttctgtgctggctggt60cttctgctgggagcctgccaggcacaccccatccctgactccagtcctctcctgcaattc120gggggccaagtccggcagcggtacctctacacagatgatgcccagcagacagaagcccac180ctggagatcagggaggatgggacggtggggggcgctgctgaccagagccccgaaagtctc240ctgcagctgaaagccttgaagccgggagttattcaaatcttgggagtcaagacatccagg300ttcctgtgccagcggccagatggggccctgtatggatcgctccactttgaccctgaggcc360tgcagcttccgggagctgcttcttgaggacggatacaatgtttaccagtccgaagcccac420ggcctcccgctgcacctgccagggaacaagtccccacaccgggaccctgcaccccgagga480ccagctcgcttcctgccactaccaggcctgccccccgcactcccggagccacccggaatc540ctggccccccagccccccgatgtgggctcctcggaccctctgagcatggtgggaccttcc600cagggccgaagccccagctacgcttcctga63014627DNA人Homosapiens14atggactcggacgagaccgggttcgagcactcaggactgtgggtttctgtgctggctggt60cttctgggagcctgccaggcacaccccatccctgactccagtcctctcctgcaattcggg120ggccaagtccggcagcggtacctctacacagatgatgcccagcagacagaagcccacctg180gagatcagggaggatgggacggtggggggcgctgctgaccagagccccgaaagtctcctg240cagctgaaagccttgaagccgggagttattcaaatcttgggagtcaagacatccaggttc300ctgtgccagcggccagatggggccctgtatggatcgctccactttgaccctgaggcctgc360agcttccgggagctgcttcttgaggacggatacaatgtttaccagtccgaagcccacggc420ctcccgctgcacctgccagggaacaagtccccacaccgggaccctgcaccccgaggacca480gctcgcttcctgccactaccaggcctgccccccgcactcccggagccacccggaatcctg540gccccccagccccccgatgtgggctcctcggaccctctgagcatggtgggaccttcccag600ggccgaagccccagctacgcttcctga6271577DNA人工衍生自詹氏甲烷球菌MethanococcusjannaschiitRNA的突变体tRNA15ccggcggtagttcagcagggcagaacggcggactctaaatccgcatggcgctggttcaaa60tccagcccgccggacca7716306PRT人工衍生自詹氏甲烷球菌Methanococcusjannaschii合成酶的突变体合成酶16MetAspGluPheGluMetIleLysArgAsnThrSerGluIleIleSer151015GluGluGluLeuArgGluValLeuLysLysAspGluLysSerAlaVal202530IleGlyPheGluProSerGlyLysIleHisLeuGlyHisTyrLeuGln354045IleLysLysMetIleAspLeuGlnAsnAlaGlyPheAspIleIleIle505560TyrLeuAlaAspLeuHisAlaTyrLeuAsnGlnLysGlyGluLeuAsp65707580GluIleArgLysIleGlyAspTyrAsnLysLysValPheGluAlaMet859095GlyLeuLysAlaLysTyrValTyrGlySerGluHisGlyLeuAspLys100105110AspTyrThrLeuAsnValTyrArgLeuAlaLeuLysThrThrLeuLys115120125ArgAlaArgArgSerMetGluLeuIleAlaArgGluAspGluAsnPro130135140LysValAlaGluValIleTyrProIleMetGlnValAsnGlyIleHis145150155160TyrGluGlyValAspValAlaValGlyGlyMetGluGlnArgLysIle165170175HisMetLeuAlaArgGluLeuLeuProLysLysValValCysIleHis180185190AsnProValLeuThrGlyLeuAspGlyGluGlyLysMetSerSerSer195200205LysGlyAsnPheIleAlaValAspAspSerProGluGluIleArgAla210215220LysIleLysLysAlaTyrCysProAlaGlyValValGluGlyAsnPro225230235240IleMetGluIleAlaLysTyrPheLeuGluTyrProLeuThrIleLys245250255ArgProGluLysPheGlyGlyAspLeuThrValAsnSerTyrGluGlu260265270LeuGluSerLeuPheLysAsnLysGluLeuHisProMetAspLeuLys275280285AsnAlaValAlaGluGluLeuIleLysIleLeuGluProIleArgLys290295300ArgLeu3051777DNAMethanococcusjannaschii17ccggcggtagttcagcagggcagaacggcggactctaaatccgcatggcgctggttcaaa60tccggcccgccggacca771888DNA人工优化的琥珀抑制子tRNA18cccagggtagccaagctcggccaacggcgacggactctaaatccgttctcgtaggagttc60gagggttcgaatcccttccctgggacca881989DNA人工AnoptimizedAGGAframeshiftsupressortRNA19gcgagggtagccaagctcggccaacggcgacggacttcctaatccgttctcgtaggagtt60cgagggttcgaatccctcccctcgcacca8920306PRT人工用于并入p-叠氮-L-苯丙氨酸的氨基酰tRNA合成酶20MetAspGluPheGluMetIleLysArgAsnThrSerGluIleIleSer151015GluGluGluLeuArgGluValLeuLysLysAspGluLysSerAlaGly202530IleGlyPheGluProSerGlyLysIleHisLeuGlyHisTyrLeuGln354045IleLysLysMetIleAspLeuGlnAsnAlaGlyPheAspIleIleIle505560LeuLeuAlaAspLeuHisAlaTyrLeuAsnGlnLysGlyGluLeuAsp65707580GluIleArgLysIleGlyAspTyrAsnLysLysValPheGluAlaMet859095GlyLeuLysAlaLysTyrValTyrGlySerThrPheGlnLeuAspLys100105110AspTyrThrLeuAsnValTyrArgLeuAlaLeuLysThrThrLeuLys115120125ArgAlaArgArgSerMetGluLeuIleAlaArgGluAspGluAsnPro130135140LysValAlaGluValIleTyrProIleMetGlnValAsnThrTyrTyr145150155160TyrLeuGlyValAspValAlaValGlyGlyMetGluGlnArgLysIle165170175HisMetLeuAlaArgGluLeuLeuProLysLysValValCysIleHis180185190AsnProValLeuThrGlyLeuAspGlyGluGlyLysMetSerSerSer195200205LysGlyAsnPheIleAlaValAspAspSerProGluGluIleArgAla210215220LysIleLysLysAlaTyrCysProAlaGlyValValGluGlyAsnPro225230235240IleMetGluIleAlaLysTyrPheLeuGluTyrProLeuThrIleLys245250255ArgProGluLysPheGlyGlyAspLeuThrValAsnSerTyrGluGlu260265270LeuGluSerLeuPheLysAsnLysGluLeuHisProMetAspLeuLys275280285AsnAlaValAlaGluGluLeuIleLysIleLeuGluProIleArgLys290295300ArgLeu30521306PRT人工用于并入p-苯甲酰-L-苯丙氨酸的氨基酰tRNA合成酶21MetAspGluPheGluMetIleLysArgAsnThrSerGluIleIleSer151015GluGluGluLeuArgGluValLeuLysLysAspGluLysSerAlaGly202530IleGlyPheGluProSerGlyLysIleHisLeuGlyHisTyrLeuGln354045IleLysLysMetIleAspLeuGlnAsnAlaGlyPheAspIleIleIle505560LeuLeuAlaAspLeuHisAlaTyrLeuAsnGlnLysGlyGluLeuAsp65707580GluIleArgLysIleGlyAspTyrAsnLysLysValPheGluAlaMet859095GlyLeuLysAlaLysTyrValTyrGlySerSerPheGlnLeuAspLys100105110AspTyrThrLeuAsnValTyrArgLeuAlaLeuLysThrThrLeuLys115120125ArgAlaArgArgSerMetGluLeuIleAlaArgGluAspGluAsnPro130135140LysValAlaGluValIleTyrProIleMetGlnValAsnThrSerHis145150155160TyrLeuGlyValAspValAlaValGlyGlyMetGluGlnArgLysIle165170175HisMetLeuAlaArgGluLeuLeuProLysLysValValCysIleHis180185190AsnProValLeuThrGlyLeuAspGlyGluGlyLysMetSerSerSer195200205LysGlyAsnPheIleAlaValAspAspSerProGluGluIleArgAla210215220LysIleLysLysAlaTyrCysProAlaGlyValValGluGlyAsnPro225230235240IleMetGluIleAlaLysTyrPheLeuGluTyrProLeuThrIleLys245250255ArgProGluLysPheGlyGlyAspLeuThrValAsnSerTyrGluGlu260265270LeuGluSerLeuPheLysAsnLysGluLeuHisProMetAspLeuLys275280285AsnAlaValAlaGluGluLeuIleLysIleLeuGluProIleArgLys290295300ArgLeu30522305PRT人工用于并入炔丙基-苯丙氨酸的氨基酰tRNA合成酶22MetAspGluPheGluMetIleLysArgAsnThrSerGluIleIleSer151015GluGluGluLeuArgGluValLeuLysLysAspGluLysSerAlaAla202530IleGlyPheGluProSerGlyLysIleHisLeuGlyHisTyrLeuGln354045IleLysLysMetIleAspLeuGlnAsnAlaGlyPheAspIleIleIle505560LeuLeuAlaAspLeuHisAlaTyrLeuAsnGlnLysGlyGluLeuAsp65707580GluIleArgLysIleGlyAspTyrAsnLysLysValPheGluAlaMet859095GlyLeuLysAlaLysTyrValTyrGlySerProPheGlnLeuAspLys100105110AspTyrThrLeuAsnValTyrArgLeuAlaLeuLysThrThrLeuLys115120125ArgAlaArgArgSerMetGluLeuIleAlaArgGluAspGluAsnPro130135140LysValAlaGluValIleTyrProIleMetGlnValAsnAlaIleTyr145150155160LeuAlaValAspValAlaValGlyGlyMetGluGlnArgLysIleHis165170175MetLeuAlaArgGluLeuLeuProLysLysValValCysIleHisAsn180185190ProValLeuThrGlyLeuAspGlyGluGlyLysMetSerSerSerLys195200205GlyAsnPheIleAlaValAspAspSerProGluGluIleArgAlaLys210215220IleLysLysAlaTyrCysProAlaGlyValValGluGlyAsnProIle225230235240MetGluIleAlaLysTyrPheLeuGluTyrProLeuThrIleLysArg245250255ProGluLysPheGlyGlyAspLeuThrValAsnSerTyrGluGluLeu260265270GluSerLeuPheLysAsnLysGluLeuHisProMetAspLeuLysAsn275280285AlaValAlaGluGluLeuIleLysIleLeuGluProIleArgLysArg290295300Leu30523305PRT人工用于并入炔丙基-苯丙氨酸的氨基酰tRNA合成酶23MetAspGluPheGluMetIleLysArgAsnThrSerGluIleIleSer151015GluGluGluLeuArgGluValLeuLysLysAspGluLysSerAlaAla202530IleGlyPheGluProSerGlyLysIleHisLeuGlyHisTyrLeuGln354045IleLysLysMetIleAspLeuGlnAsnAlaGlyPheAspIleIleIle505560LeuLeuAlaAspLeuHisAlaTyrLeuAsnGlnLysGlyGluLeuAsp65707580GluIleArgLysIleGlyAspTyrAsnLysLysValPheGluAlaMet859095GlyLeuLysAlaLysTyrValTyrGlySerProPheGlnLeuAspLys100105110AspTyrThrLeuAsnValTyrArgLeuAlaLeuLysThrThrLeuLys115120125ArgAlaArgArgSerMetGluLeuIleAlaArgGluAspGluAsnPro130135140LysValAlaGluValIleTyrProIleMetGlnValAsnIleProTyr145150155160LeuProValAspValAlaValGlyGlyMetGluGlnArgLysIleHis165170175MetLeuAlaArgGluLeuLeuProLysLysValValCysIleHisAsn180185190ProValLeuThrGlyLeuAspGlyGluGlyLysMetSerSerSerLys195200205GlyAsnPheIleAlaValAspAspSerProGluGluIleArgAlaLys210215220IleLysLysAlaTyrCysProAlaGlyValValGluGlyAsnProIle225230235240MetGluIleAlaLysTyrPheLeuGluTyrProLeuThrIleLysArg245250255ProGluLysPheGlyGlyAspLeuThrValAsnSerTyrGluGluLeu260265270GluSerLeuPheLysAsnLysGluLeuHisProMetAspLeuLysAsn275280285AlaValAlaGluGluLeuIleLysIleLeuGluProIleArgLysArg290295300Leu30524305PRT人工用于并入炔丙基-苯丙氨酸的氨基酰tRNA合成酶24MetAspGluPheGluMetIleLysArgAsnThrSerGluIleIleSer151015GluGluGluLeuArgGluValLeuLysLysAspGluLysSerAlaAla202530IleGlyPheGluProSerGlyLysIleHisLeuGlyHisTyrLeuGln354045IleLysLysMetIleAspLeuGlnAsnAlaGlyPheAspIleIleIle505560LeuLeuAlaAspLeuHisAlaTyrLeuAsnGlnLysGlyGluLeuAsp65707580GluIleArgLysIleGlyAspTyrAsnLysLysValPheGluAlaMet859095GlyLeuLysAlaLysTyrValTyrGlySerLysPheGlnLeuAspLys100105110AspTyrThrLeuAsnValTyrArgLeuAlaLeuLysThrThrLeuLys115120125ArgAlaArgArgSerMetGluLeuIleAlaArgGluAspGluAsnPro130135140LysValAlaGluValIleTyrProIleMetGlnValAsnAlaIleTyr145150155160LeuAlaValAspValAlaValGlyGlyMetGluGlnArgLysIleHis165170175MetLeuAlaArgGluLeuLeuProLysLysValValCysIleHisAsn180185190ProValLeuThrGlyLeuAspGlyGluGlyLysMetSerSerSerLys195200205GlyAsnPheIleAlaValAspAspSerProGluGluIleArgAlaLys210215220IleLysLysAlaTyrCysProAlaGlyValValGluGlyAsnProIle225230235240MetGluIleAlaLysTyrPheLeuGluTyrProLeuThrIleLysArg245250255ProGluLysPheGlyGlyAspLeuThrValAsnSerTyrGluGluLeu260265270GluSerLeuPheLysAsnLysGluLeuHisProMetAspLeuLysAsn275280285AlaValAlaGluGluLeuIleLysIleLeuGluProIleArgLysArg290295300Leu30525306PRT人工用于并入p-叠氮-苯丙氨酸的氨基酰tRNA合成酶25MetAspGluPheGluMetIleLysArgAsnThrSerGluIleIleSer151015GluGluGluLeuArgGluValLeuLysLysAspGluLysSerAlaThr202530IleGlyPheGluProSerGlyLysIleHisLeuGlyHisTyrLeuGln354045IleLysLysMetIleAspLeuGlnAsnAlaGlyPheAspIleIleIle505560LeuLeuAlaAspLeuHisAlaTyrLeuAsnGlnLysGlyGluLeuAsp65707580GluIleArgLysIleGlyAspTyrAsnLysLysValPheGluAlaMet859095GlyLeuLysAlaLysTyrValTyrGlySerAsnPheGlnLeuAspLys100105110AspTyrThrLeuAsnValTyrArgLeuAlaLeuLysThrThrLeuLys115120125ArgAlaArgArgSerMetGluLeuIleAlaArgGluAspGluAsnPro130135140LysValAlaGluValIleTyrProIleMetGlnValAsnProLeuHis145150155160TyrGlnGlyValAspValAlaValGlyGlyMetGluGlnArgLysIle165170175HisMetLeuAlaArgGluLeuLeuProLysLysValValCysIleHis180185190AsnProValLeuThrGlyLeuAspGlyGluGlyLysMetSerSerSer195200205LysGlyAsnPheIleAlaValAspAspSerProGluGluIleArgAla210215220LysIleLysLysAlaTyrCysProAlaGlyValValGluGlyAsnPro225230235240IleMetGluIleAlaLysTyrPheLeuGluTyrProLeuThrIleLys245250255ArgProGluLysPheGlyGlyAspLeuThrValAsnSerTyrGluGlu260265270LeuGluSerLeuPheLysAsnLysGluLeuHisProMetAspLeuLys275280285AsnAlaValAlaGluGluLeuIleLysIleLeuGluProIleArgLys290295300ArgLeu30526306PRT人工用于并入p-叠氮-苯丙氨酸的氨基酰tRNA合成酶26MetAspGluPheGluMetIleLysArgAsnThrSerGluIleIleSer151015GluGluGluLeuArgGluValLeuLysLysAspGluLysSerAlaThr202530IleGlyPheGluProSerGlyLysIleHisLeuGlyHisTyrLeuGln354045IleLysLysMetIleAspLeuGlnAsnAlaGlyPheAspIleIleIle505560LeuLeuAlaAspLeuHisAlaTyrLeuAsnGlnLysGlyGluLeuAsp65707580GluIleArgLysIleGlyAspTyrAsnLysLysValPheGluAlaMet859095GlyLeuLysAlaLysTyrValTyrGlySerSerPheGlnLeuAspLys100105110AspTyrThrLeuAsnValTyrArgLeuAlaLeuLysThrThrLeuLys115120125ArgAlaArgArgSerMetGluLeuIleAlaArgGluAspGluAsnPro130135140LysValAlaGluValIleTyrProIleMetGlnValAsnProLeuHis145150155160TyrGlnGlyValAspValAlaValGlyGlyMetGluGlnArgLysIle165170175HisMetLeuAlaArgGluLeuLeuProLysLysValValCysIleHis180185190AsnProValLeuThrGlyLeuAspGlyGluGlyLysMetSerSerSer195200205LysGlyAsnPheIleAlaValAspAspSerProGluGluIleArgAla210215220LysIleLysLysAlaTyrCysProAlaGlyValValGluGlyAsnPro225230235240IleMetGluIleAlaLysTyrPheLeuGluTyrProLeuThrIleLys245250255ArgProGluLysPheGlyGlyAspLeuThrValAsnSerTyrGluGlu260265270LeuGluSerLeuPheLysAsnLysGluLeuHisProMetAspLeuLys275280285AsnAlaValAlaGluGluLeuIleLysIleLeuGluProIleArgLys290295300ArgLeu30527306PRT人工用于并入p-叠氮-苯丙氨酸的氨基酰tRNA合成酶27MetAspGluPheGluMetIleLysArgAsnThrSerGluIleIleSer151015GluGluGluLeuArgGluValLeuLysLysAspGluLysSerAlaLeu202530IleGlyPheGluProSerGlyLysIleHisLeuGlyHisTyrLeuGln354045IleLysLysMetIleAspLeuGlnAsnAlaGlyPheAspIleIleIle505560LeuLeuAlaAspLeuHisAlaTyrLeuAsnGlnLysGlyGluLeuAsp65707580GluIleArgLysIleGlyAspTyrAsnLysLysValPheGluAlaMet859095GlyLeuLysAlaLysTyrValTyrGlySerThrPheGlnLeuAspLys100105110AspTyrThrLeuAsnValTyrArgLeuAlaLeuLysT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60TyrAlaValSerLysAspAspTyrTyrProHisGluHisArgProIle65707580SerIleAsnTyrArgThr855584PRT人工序列合成多肽55GluValValAlaAlaThrProThrSerLeuLeuIleSerTrpAspAla151015SerHisTyrGluArgArgTyrTyrArgIleThrTyrGlyGluThrGly202530GlyAsnSerProValGlnGluPheThrValProArgTyrHisHisThr354045AlaThrIleSerGlyLeuLysProGlyValAspTyrThrIleThrVal505560TyrAlaValThrGlnAlaGlnGluHisTyrGlnProProIleSerIle65707580AsnTyrArgThr5682PRT人工序列合成多肽56GluValValAlaAlaThrProThrSerLeuLeuIleSerTrpAsnSer151015TyrTyrHisSerAlaAspTyrTyrArgIleThrTyrGlyGluThrGly202530GlyAsnSerProValGlnGluPheThrValProTyrProProThrThr354045AlaThrIleSerGlyLeuLysProGlyValAspTyrThrIleThrVal505560TyrAlaValTyrSerAlaLysSerTyrTyrProIleSerIleAsnTyr65707580ArgThr5786PRT人工序列合成多肽57GluValValAlaAlaThrProThrSerLeuLeuIleSerTrpSerLys151015TyrSerLysHisGlyHisTyrTyrArgIleThrTyrGlyGluThrGly202530GlyAsnSerProValGlnGluPheThrValProSerGlyAsnAlaThr354045AlaThrIleSerGlyLeuLysProGlyValAspTyrThrIleThrVal505560TyrAlaValGluAspThrAsnAspTyrProHisThrHisArgProIle65707580SerIleAsnTyrArgThr855884PRT人工序列合成多肽58GluValValAlaAlaThrProThrSerLeuLeuIleSerTrpHisGly151015GluProAspGlnThrArgTyrTyrArgIleThrTyrGlyGluThrGly202530GlyAsnSerProValGlnGluPheThrValProProTyrArgArgThr354045AlaThrIleSerGlyLeuLysProGlyValAspTyrThrIleThrVal505560TyrAlaValThrSerGlyTyrThrGlyHisTyrGlnProIleSerIle65707580AsnTyrArgThr5986PRT人工序列合成多肽59GluValValAlaAlaThrProThrSerLeuLeuIleSerTrpSerLys151015TyrSerLysHisGlyHisTyrTyrArgIleThrTyrGlyGluThrGly202530GlyAsnSerProValGlnGluPheThrValAspProSerSerTyrThr354045AlaThrIleSerGlyLeuLysProGlyValAspTyrThrIleThrVal505560TyrAlaValSerLysAspAspTyrTyrProHisGluHisArgProIle65707580SerIleAsnTyrArgThr856082PRT人工序列合成多肽60GluValValAlaAlaThrProThrSerLeuLeuIleSerTrpTyrGlu151015ProTyrThrProIleHisTyrTyrArgIleThrTyrGlyGluThrGly202530GlyAsnSerProValGlnGluPheThrValProGlyTyrTyrGlyThr354045AlaThrIleSerGlyLeuLysProGlyValAspTyrThrIleThrVal505560TyrAlaValTyrGlyTyrTyrGlnTyrThrProIleSerIleAsnTyr65707580ArgThr6186PRT人工序列合成多肽61GluValValAlaAlaThrProThrSerLeuLeuIleSerTrpSerLys151015TyrSerLysHisGlyHisTyrTyrArgIleThrTyrGlyGluThrGly202530GlyAsnSerProValGlnGluPheThrValProSerGlyAsnAlaThr354045AlaThrIleSerGlyLeuLysProGlyValAspTyrThrIleThrVal505560TyrAlaValSerAspAspAsnLysTyrTyrHisGlnHisArgProIle65707580SerIleAsnTyrArgThr856286PRT人工序列合成多肽62GluValValAlaAlaThrProThrSerLeuLeuIleSerTrpGlyHis151015TyrArgArgSerGlyHisTyrTyrArgIleThrTyrGlyGluThrGly202530GlyAsnSerProValGlnGluPheThrValAspProSerSerTyrThr354045AlaThrIleSerGlyLeuLysProGlyValAspTyrThrIleThrVal505560TyrAlaValSerLysAspAspTyrTyrProHisGluHisArgProIle65707580SerIleAsnTyrArgThr856386PRT人工序列合成多肽63GluValValAlaAlaThrProThrSerLeuLeuIleSerTrpSerLys151015TyrSerLysHisGlyHisTyrTyrArgIleThrTyrGlyGluThrGly202530GlyAsnSerProValGlnGluPheThrValProSerGlyAsnAlaThr354045AlaThrIleSerGlyLeuLysProGlyValAspTyrThrIleThrVal505560TyrAlaValGluAspThrAsnAspTyrProHisThrHisArgProIle65707580SerIleAsnTyrArgThr856482PRT人工序列合成多肽64GluValValAlaAlaThrProThrSerLeuLeuIleSerTrpTyrGlu151015ProGlyAlaSerValTyrTyrTyrArgIleThrTyrGlyGluThrGly202530GlyAsnSerProValGlnGluPheThrValProSerTyrTyrHisThr354045AlaThrIleSerGlyLeuLysProGlyValAspTyrThrIleThrVal505560TyrAlaValTyrGlyTyrTyrGluTyrGluProIleSerIleAsnTyr65707580ArgThr6582PRT人工序列合成多肽65GluValValAlaAlaThrProThrSerLeuLeuIleSerTrpGlnSer151015TyrTyrAlaHisSerAspTyrTyrArgIleThrTyrGlyGluThrGly202530GlyAsnSerProValGlnGluPheThrValProTyrProProGlnThr354045AlaThrIleSerGlyLeuLysProGlyValAspTyrThrIleThrVal505560TyrAlaValTyrAlaGlySerSerTyrTyrProIleSerIleAsnTyr65707580ArgThr6686PRT人工序列合成多肽66GluValValAlaAlaThrProThrSerLeuLeuIleSerTrpGlyHis151015TyrArgArgSerGlyHisTyrTyrArgIleThrTyrGlyGluThrGly202530GlyAsnSerProValGlnGluPheThrValAspProSerSerTyrThr354045AlaThrIleSerGlyLeuLysProGlyValAspTyrThrIleThrVal505560TyrAlaValSerLysAspAspTyrTyrProHisGluHisArgProIle65707580SerIleAsnTyrArgThr856782PRT人工序列合成多肽67GluValValAlaAlaThrProThrSerLeuLeuIleSerTrpProGlu151015ProGlyThrProValTyrTyrTyrArgIleThrTyrGlyGluThrGly202530GlyAsnSerProValGlnGluPheThrValProAlaTyrTyrGlyThr354045AlaThrIleSerGlyLeuLysProGlyValAspTyrThrIleThrVal505560TyrAlaValTyrGlyTyrTyrAspTyrSerProIleSerIleAsnTyr65707580ArgThr6886PRT人工序列合成多肽68GluValValAlaAlaThrProThrSerLeuLeuIleSerTrpTyrArg151015TyrGluLysThrGlnHisTyrTyrArgIleThrTyrGlyGluThrGly202530GlyAsnSerProValGlnGluPheThrValProProGluSerGlyThr354045AlaThrIleSerGlyLeuLysProGlyValAspTyrThrIleThrVal505560TyrAlaValTyrAlaGlyTyrGluTyrProHisThrHisArgProIle65707580SerIleAsnTyrArgThr856987PRT人工序列合成多肽69GluValValAlaAlaThrProThrSerLeuLeuIleSerTrpValLys151015SerGluGluTyrTyrArgTyrTyrArgIleThrTyrGlyGluThrGly202530GlyAsnSerProValGlnGluPheThrValProTyrTyrValHisThr354045AlaThrIleSerGlyLeuLysProGlyValAspTyrThrIleThrVal505560TyrAlaValThrGluTyrTyrTyrAlaGlyAlaValValSerValPro65707580IleSerIleAsnTyrArgThr857082PRT人工序列合成多肽70GluValValAlaAlaThrProThrSerLeuLeuIleSerTrpTyrAsp151015ProTyrThrTyrGlySerTyrTyrArgIleThrTyrGlyGluThrGly202530GlyAsnSerProValGlnGluPheThrValGlyProTyrThrThrThr354045AlaThrIleSerGlyLeuLysProGlyValAspTyrThrIleThrVal505560TyrAlaValSerTyrTyrTyrSerThrGlnProIleSerIleAsnTyr65707580ArgThr7186PRT人工序列合成多肽71GluValValAlaAlaThrProThrSerLeuLeuIleSerTrpSerAsn151015AspGlyProGlyLeuSerTyrTyrArgIleThrTyrGlyGluThrGly202530GlyAsnSerProValGlnGluPheThrValProSerSerGlnThrThr354045AlaThrIleSerGlyLeuLysProGlyValAspTyrThrIleThrVal505560TyrAlaValSerTyrTyrThrLysLysAlaTyrSerAlaGlyProIle65707580SerIleAsnTyrArgThr857282PRT人工序列合成多肽72GluValValAlaAlaThrProThrSerLeuLeuIleSerTrpProAsp151015ProTyrTyrLysProAspTyrTyrArgIleThrTyrGlyGluThrGly202530GlyAsnSerProValGlnGluPheThrValProArgAspTyrThrThr354045AlaThrIleSerGlyLeuLysProGlyValAspTyrThrIleThrVal505560TyrAlaValTyrSerTyrTyrGlyTyrTyrProIleSerIleAsnTyr65707580ArgThr7312PRT人工合成序列73ProGlyAsnLysSerProHisArgAspProAlaPro1510748PRT人工合成序列74GlyAlaGlyGlyGlyGlySerGly15756PRT人工合成序列75GluProLysSerSerAsp157621PRT人工合成序列76GluSerProLysAlaGlnAlaSerSerValProThrAlaGlnProGln151015AlaGluGlyLeuAla207717PRT人工合成序列77GluLeuGlnLeuGluGluSerAlaAlaGluAlaGlnAspGlyGluLeu151015Asp789PRT人工合成序列78GlyGlnProAspGluProGlyGlySer157913PRT人工合成序列79GlyGlySerGlySerGlySerGlySerGlySerGlySer15108017PRT人工合成序列80GluLeuGlnLeuGluGluSerAlaAlaGluAlaGlnGluGlyGluLeu151015Glu815PRT人工合成序列81GlySerGlySerGly15824PRT人工合成序列82GlySerGlyCys1837PRT人工合成序列83AlaGlyGlyGlyGlySerGly15844PRT人工合成序列84GlySerGlySer1858PRT人工合成序列85GlnProAspGluProGlyGlySer15866PRT人工合成序列86GlySerGlySerGlySer15876PRT人工合成序列87ThrValAlaAlaProSer15888PRT人工合成序列88LysAlaGlyGlyGlyGlySerGly158913PRT人工合成序列89LysGlySerGlySerGlySerGlySerGlySerGlySer1510909PRT人工合成序列90LysGlnProAspGluProGlyGlySer159118PRT人工合成序列91LysGluLeuGlnLeuGluGluSerAlaAlaGluAlaGlnAspGlyGlu151015LeuAsp927PRT人工合成序列92LysThrValAlaAlaProSer15939PRT人工合成序列93LysAlaGlyGlyGlyGlySerGlyGly159414PRT人工合成序列94LysGlySerGlySerGlySerGlySerGlySerGlySerGly15109510PRT人工合成序列95LysGlnProAspGluProGlyGlySerGly15109619PRT人工合成序列96LysGluLeuGlnLeuGluGluSerAlaAlaGluAlaGlnAspGlyGlu151015LeuAspGly9716PRT人工合成序列97LysThrValAlaAlaProSerGlyAlaGlyGlyGlyGlySerGlyGly151015987PRT人工合成序列98AlaGlyGlyGlyGlySerGly159913PRT人工合成序列99GlySerGlySerGlySerGlySerGlySerGlySerGly15101009PRT人工合成序列100GlnProAspGluProGlyGlySerGly15101182PRT人工合成序列101MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrGlnSerGluAla100105110HisGlyLeuProLeuHisLeuProGlyAsnLysSerProHisArgAsp115120125ProAlaProArgGlyProAlaArgPheLeuProLeuProGlyLeuPro130135140ProAlaProProGluProProGlyIleLeuAlaProGlnProProAsp145150155160ValGlySerSerAspProLeuSerMetValGlyProSerGlnGlyArg165170175SerProSerTyrAlaSer180102173PRT人工合成序列102MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrGlnSerGluAla100105110HisGlyLeuProLeuHisLeuGlySerGlyArgGlyProAlaArgPhe115120125LeuProLeuProGlyLeuProProAlaProProGluProProGlyIle130135140LeuAlaProGlnProProAspValGlySerSerAspProLeuSerMet145150155160ValGluProSerGlnGlyArgSerProSerTyrAlaSer165170103182PRT人工合成序列103MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrGlnSerGluAla100105110HisGlyLeuProLeuHisLeuProGlyAsnLysSerProHisArgAsp115120125ProAlaProArgGlyProAlaArgPheLeuProLeuProGlyLeuPro130135140ProAlaProProGluProProGlyIleLeuAlaProGlnProProAsp145150155160ValGlySerSerAspProLeuSerMetValGluProSerGlnGlyArg165170175SerProSerTyrAlaSer180104182PRT人工合成序列104MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrGlnSerGluAla100105110HisGlyLeuProLeuHisLeuProGlyLysLysSerProHisArgAsp115120125ProAlaProArgGlyProAlaArgPheLeuProLeuProGlyLeuPro130135140ProAlaProProGluProProGlyIleLeuAlaProGlnProProAsp145150155160ValGlySerSerAspProLeuSerMetValGluProSerGlnGlyArg165170175SerProSerTyrAlaSer180105180PRT人工合成序列105MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrGlnSerGluAla100105110HisGlyLeuProLeuHisLeuProGlyAspLysSerArgAspProAla115120125ProArgGlyProAlaArgPheLeuProLeuProGlyLeuProProAla130135140ProProGluProProGlyIleLeuAlaProGlnProProAspValGly145150155160SerSerAspProLeuSerMetValGluProSerGlnGlyArgSerPro165170175SerTyrAlaSer180106180PRT人工合成序列106MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrGlnSerGluAla100105110HisGlyLeuProLeuHisLeuProGlyHisLysSerArgAspProAla115120125ProArgGlyProAlaArgPheLeuProLeuProGlyLeuProProAla130135140ProProGluProProGlyIleLeuAlaProGlnProProAspValGly145150155160SerSerAspProLeuSerMetValGluProSerGlnGlyArgSerPro165170175SerTyrAlaSer180107182PRT人工合成序列107MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrGlnSerGluAla100105110HisGlyLeuProLeuHisLeuProGlyAspLysSerProHisArgAsp115120125ProAlaProArgGlyProAlaArgPheLeuProLeuProGlyLeuPro130135140ProAlaProProGluProProGlyIleLeuAlaProGlnProProAsp145150155160ValGlySerSerAspProLeuSerMetValGluProSerGlnGlyArg165170175SerProSerTyrAlaSer180108170PRT人工合成序列108MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrGlnSerGluAla100105110HisGlyLeuProLeuHisLeuGlySerGlyAlaArgPheLeuProLeu115120125ProGlyLeuProProAlaProProGluProProGlyIleLeuAlaPro130135140GlnProProAspValGlySerSerAspProLeuSerMetValGluPro145150155160SerGlnGlyArgSerProSerTyrAlaSer165170109182PRT人工合成序列109MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrGlnSerGluAla100105110HisGlyLeuProLeuHisLeuProGlyGlnLysSerProHisArgAsp115120125ProAlaProArgGlyProAlaArgPheLeuProLeuProGlyLeuPro130135140ProAlaProProGluProProGlyIleLeuAlaProGlnProProAsp145150155160ValGlySerSerAspProLeuSerMetValGluProSerGlnGlyArg165170175SerProSerTyrAlaSer180110172PRT人工合成序列110MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrGlnSerGluAla100105110HisGlyLeuProLeuHisLeuGlySerGlyGlyProAlaArgPheLeu115120125ProLeuProGlyLeuProProAlaProProGluProProGlyIleLeu130135140AlaProGlnProProAspValGlySerSerAspProLeuSerMetVal145150155160GluProSerGlnGlyArgSerProSerTyrAlaSer165170111179PRT人工合成序列111MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrGlnSerGluAla100105110HisGlyLeuProLeuHisLeuGlySerGlyHisArgAspProAlaPro115120125ArgGlyProAlaArgPheLeuProLeuProGlyLeuProProAlaPro130135140ProGluProProGlyIleLeuAlaProGlnProProAspValGlySer145150155160SerAspProLeuSerMetValGluProSerGlnGlyArgSerProSer165170175TyrAlaSer112180PRT人工合成序列112MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrGlnSerGluAla100105110HisGlyLeuProLeuHisLeuProHisHisSerGlyArgAspProAla115120125ProArgGlyProAlaArgPheLeuProLeuProGlyLeuProProAla130135140ProProGluProProGlyIleLeuAlaProGlnProProAspValGly145150155160SerSerAspProLeuSerMetValGluProSerGlnGlyArgSerPro165170175SerTyrAlaSer180113180PRT人工合成序列113MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrGlnSerGluAla1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HisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrGlnSerGluAla100105110HisGlyLeuProHisSerGlyGlyArgPheLeuProLeuProGlyLeu115120125ProProAlaProProGluProProGlyIleLeuAlaProGlnProPro130135140AspValGlySerSerAspProLeuSerMetValGluProSerGlnGly145150155160ArgSerProSerTyrAlaSer165130167PRT人工合成序列130MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrGlnSerGluAla100105110HisGlyLeuProHisGlySerGlyArgPheLeuProLeuProGlyLeu115120125ProProAlaProProGluProProGlyIleLeuAlaProGlnProPro130135140AspValGlySerSerAspProLeuSerMetValGluProSerGlnGly145150155160ArgSerProSerTyrAlaSer165131172PRT人工合成序列131MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrGlnSerGluAla100105110HisGlyLeuProLeuHisLeuGlySerGlyGlyProAlaArgPheLeu115120125ProLeuProGlyLeuProProAlaProProGluProProGlyIleLeu130135140AlaProGlnProProAspValGlySerSerAspProLeuSerMetVal145150155160ThrProSerGlnGlyArgSerProSerTyrAlaSer165170132187PRT人工合成序列132MetHisHisHisHisHisHisProIleProAspSerSerProLeuLeu151015GlnPheGlyGlyGlnValArgGlnArgTyrLeuTyrThrAspAspAla202530GlnGlnThrGluAlaHisLeuGluIleArgGluAspGlyThrValGly354045GlyAlaAlaAspGlnSerProGluSerLeuLeuGlnLeuLysAlaLeu505560LysProGlyValIleGlnIleLeuGlyValLysThrSerArgPheLeu65707580CysGlnArgProAspGlyAlaLeuTyrGlySerLeuHisPheAspPro859095GluAlaCysSerPheArgGluLeuLeuLeuGluAspGlyTyrAsnVal100105110TyrGlnSerGluAlaHisGlyLeuProLeuHisLeuProGlyAsnLys115120125SerProHisArgAspProAlaProArgGlyProAlaArgPheLeuPro130135140LeuProGlyLeuProProAlaLeuProGluProProGlyIleLeuAla145150155160ProGlnProProAspValGlySerSerAspProLeuSerMetValGly165170175ProSerGlnGlyArgSerProSerTyrAlaSer180185133133000134134000135135000136136000137137000138138000139139000140140000141141000142142000143143000144144000145145000146146000147147000148148000149149000150150000151151000152152000153153000154154000155155000156156000157157000158158000159159000160160000161161000162162000163163000164164000165165000166166000167167000168168000169169000170170000171171000172172000173173000174174000175175000176176000177177000178178000179179000180180000181181000182182000183183000184184000185185000186186000187187000188188000189189000190190000191191000192192000193193000194194000195195000196196000197197000198198000199199000200200000201182PRT人工合成序列misc_feature109..109对-乙酰基-L-苯丙氨酸，其任选连接至30kDa聚乙二醇201MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrXaaSerGluAla100105110HisGlyLeuProLeuHisLeuProGlyAsnLysSerProHisArgAsp115120125ProAlaProArgGlyProAlaArgPheLeuProLeuProGlyLeuPro130135140ProAlaProProGluProProGlyIleLeuAlaProGlnProProAsp145150155160ValGlySerSerAspProLeuSerMetValGlyProSerGlnGlyArg165170175SerProSerTyrAlaSer180202173PRT人工合成序列misc_feature109..109对-乙酰基-L-苯丙氨酸，其任选连接至30kDa聚乙二醇202MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrXaaSerGluAla100105110HisGlyLeuProLeuHisLeuGlySerGlyArgGlyProAlaArgPhe115120125LeuProLeuProGlyLeuProProAlaProProGluProProGlyIle130135140LeuAlaProGlnProProAspValGlySerSerAspProLeuSerMet145150155160ValGluProSerGlnGlyArgSerProSerTyrAlaSer165170203203000204204000205180PRT人工合成序列misc_feature109..109对-乙酰基-L-苯丙氨酸，其任选连接至30kDa聚乙二醇205MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrXaaSerGluAla100105110HisGlyLeuProLeuHisLeuProGlyAspLysSerArgAspProAla115120125ProArgGlyProAlaArgPheLeuProLeuProGlyLeuProProAla130135140ProProGluProProGlyIleLeuAlaProGlnProProAspValGly145150155160SerSerAspProLeuSerMetValGluProSerGlnGlyArgSerPro165170175SerTyrAlaSer180206180PRT人工合成序列misc_feature109..109对-乙酰基-L-苯丙氨酸，其任选连接至30kDa聚乙二醇206MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrXaaSerGluAla100105110HisGlyLeuProLeuHisLeuProGlyHisLysSerArgAspProAla115120125ProArgGlyProAlaArgPheLeuProLeuProGlyLeuProProAla130135140ProProGluProProGlyIleLeuAlaProGlnProProAspValGly145150155160SerSerAspProLeuSerMetValGluProSerGlnGlyArgSerPro165170175SerTyrAlaSer180207207000208208000209209000210172PRT人工合成序列misc_feature109..109对-乙酰基-L-苯丙氨酸，其任选连接至30kDa聚乙二醇210MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrXaaSerGluAla100105110HisGlyLeuProLeuHisLeuGlySerGlyGlyProAlaArgPheLeu115120125ProLeuProGlyLeuProProAlaProProGluProProGlyIleLeu130135140AlaProGlnProProAspValGlySerSerAspProLeuSerMetVal145150155160GluProSerGlnGlyArgSerProSerTyrAlaSer165170211179PRT人工合成序列misc_feature109..109对-乙酰基-L-苯丙氨酸，其任选连接至30kDa聚乙二醇211MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrXaaSerGluAla100105110HisGlyLeuProLeuHisLeuGlySerGlyHisArgAspProAlaPro115120125ArgGlyProAlaArgPheLeuProLeuProGlyLeuProProAlaPro130135140ProGluProProGlyIleLeuAlaProGlnProProAspValGlySer145150155160SerAspProLeuSerMetValGluProSerGlnGlyArgSerProSer165170175TyrAlaSer212180PRT人工合成序列misc_feature109..109对-乙酰基-L-苯丙氨酸，其任选连接至30kDa聚乙二醇212MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrXaaSerGluAla100105110HisGlyLeuProLeuHisLeuProHisHisSerGlyArgAspProAla115120125ProArgGlyProAlaArgPheLeuProLeuProGlyLeuProProAla130135140ProProGluProProGlyIleLeuAlaProGlnProProAspValGly145150155160SerSerAspProLeuSerMetValGluProSerGlnGlyArgSerPro165170175SerTyrAlaSer180213213000214214000215215000216216000217217000218218000219180PRT人工合成序列misc_feature109..109对-乙酰基-L-苯丙氨酸，其任选连接至30kDa聚乙二醇219MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrXaaSerGluAla100105110HisGlyLeuProLeuHisLeuGlySerGlyProHisArgAspProAla115120125ProArgGlyProAlaArgPheLeuProLeuProGlyLeuProProAla130135140ProProGluProProGlyIleLeuAlaProGlnProProAspValGly145150155160SerSerAspProLeuSerMetValGluProSerGlnGlyArgSerPro165170175SerTyrAlaSer180220178PRT人工合成序列misc_feature109..109对-乙酰基-L-苯丙氨酸，其任选连接至30kDa聚乙二醇220MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrXaaSerGluAla100105110HisGlyLeuProLeuHisLeuGlyGlyHisArgAspProAlaProArg115120125GlyProAlaArgPheLeuProLeuProGlyLeuProProAlaProPro130135140GluProProGlyIleLeuAlaProGlnProProAspValGlySerSer145150155160AspProLeuSerMetValGluProSerGlnGlyArgSerProSerTyr165170175AlaSer221178PRT人工合成序列misc_feature109..109对-乙酰基-L-苯丙氨酸，其任选连接至30kDa聚乙二醇221MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGlyValIle505560GlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArgProAsp65707580GlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCysSerPhe859095ArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrXaaSerGluAla100105110HisGlyLeuProLeuHisLeuGlySerGlyArgAspProAlaProArg115120125GlyProAlaArgPheLeuProLeuProGlyLeuProProAlaProPro130135140GluProProGlyIleLeuAlaProGlnProProAspValGlySerSer145150155160AspProLeuSerMetValGluProSerGlnGlyArgSerProSerTyr165170175AlaSer222176PRT人工合成序列misc_feature109..109对-乙酰基-L-苯丙氨酸，其任选连接至30kDa聚乙二醇222MetHisProIleProAspSerSerProLeuLeuGlnPheGlyGlyGln151015ValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAla202530HisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAlaAspGln354045SerProGluSe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ValSerThrProThrLeuValGluValSerArgAsnLeuGlyLys420425430ValGlySerLysCysCysLysHisProGluAlaLysArgMetProCys435440445AlaGluAspTyrLeuSerValValLeuAsnGlnLeuCysValLeuHis450455460GluLysThrProValSerAspArgValThrLysCysCysThrGluSer465470475480LeuValAsnArgArgProCysPheSerAlaLeuGluValAspGluThr485490495TyrValProLysGluPheAsnAlaGluThrPheThrPheHisAlaAsp500505510IleCysThrLeuSerGluLysGluArgGlnIleLysLysGlnThrAla515520525LeuValGluLeuValLysHisLysProLysAlaThrLysGluGlnLeu530535540LysAlaValMetAspAspPheAlaAlaPheValGluLysCysCysLys545550555560AlaAspAspLysGluThrCysPheAlaGluGluGlyLysLysLeuVal565570575AlaAlaSerGlnAlaAlaLeuGlyLeuGlyGluThrGlySerHisPro580585590IleProAspSerSerProLeuLeuGlnPheGlyGlyGlnValArgGln595600605ArgTyrLeuTyrThrAspAspAlaGlnGlnThrGluAlaHisLeuGlu610615620IleArgGluAspGlyThrValGlyGlyAlaAlaAspGlnSerProGlu625630635640SerLeuLeuGlnLeuLysAlaLeuLysProGlyValIleGlnIleLeu645650655GlyValLysThrSerArgPheLeuCysGlnArgProAspGlyAlaLeu660665670TyrGlySerLeuHisPheAspProGluAlaCysSerPheArgGluLeu675680685LeuLeuGluAspGlyTyrAsnValTyrGlnSerGluAlaHisGlyLeu690695700ProLeuHisLeuGlySerGlyArgGlyProAlaArgPheLeuProLeu705710715720ProGlyLeuProProAlaProProGluProProGlyIleLeuAlaPro725730735GlnProProAspValGlySerSerAspProLeuSerMetValGluPro740745750SerGlnGluArgSerProSerTyrAlaSer755760427767PRT人工合成序列427AspAlaHisLysSerGluValAlaHisArgPheLysAspLeuGlyGlu151015GluAsnPheLysAlaLeuValLeuIleAlaPheAlaGlnTyrLeuGln202530GlnAlaProPheGluAspHisValLysLeuValAsnGluValThrGlu354045PheAlaLysThrCysValAlaAspGluSerAlaGluAsnCysAspLys505560SerLeuHisThrLeuPheGlyAspLysLeuCysThrValAlaThrLeu65707580ArgGluThrTyrGlyGluMetAlaAspCysCysAlaLysGlnGluPro859095GluArgAsnGluCysPheLeuGlnHisLysAspAspAsnProAsnLeu100105110ProArgLeuValArgProGluValAspValMetCysThrAlaPheHis115120125AspAsnGluGluThrPheLeuLysLysTyrLeuTyrGluIleAlaArg130135140ArgHisProTyrPheTyrAlaProGluLeuLeuPhePheAlaLysArg145150155160TyrLysAlaAlaPheThrGluCysCysGlnAlaAlaAspLysAlaAla165170175CysLeuLeuProLysLeuAspGluLeuArgAspGluGlyLysAlaSer180185190SerAlaLysGlnArgLeuLysCysAlaSerLeuGlnLysPheGlyGlu195200205ArgAlaPheLysAlaTrpAlaValAlaArgLeuSerGlnArgPhePro210215220LysAlaGluPheAlaGluValSerLysLeuValThrAspLeuThrLys225230235240ValHisThrGluCysCysHisGlyAspLeuLeuGluCysAlaAspAsp245250255ArgAlaAspLeuAlaLysTyrIleCysGluAsnGlnAspSerIleSer260265270SerLysLeuLysGluCysCysGluLysProLeuLeuGluLysSerHis275280285CysIleAlaGluValGluAsnAspGluMetProAlaAspLeuProSer290295300LeuAlaAlaAspPheValGluSerLysAspValCysLysAsnTyrAla305310315320GluAlaLysAspValPheLeuGlyMetPheLeuTyrGluTyrAlaArg325330335ArgHisProAspTyrSerValValLeuLeuLeuArgLeuAlaLysThr340345350TyrGluThrThrLeuGluLysCysCysAlaAlaAlaAspProHisGlu355360365CysTyrAlaLysValPheAspGluPheLysProLeuValGluGluPro370375380GlnAsnLeuIleLysGlnAsnCysGluLeuPheGluGlnLeuGlyGlu385390395400TyrLysPheGlnAsnAlaLeuLeuValArgTyrThrLysLysValPro405410415GlnValSerThrProThrLeuValGluValSerArgAsnLeuGlyLys420425430ValGlySerLysCysCysLysHisProGluAlaLysArgMetProCys435440445AlaGluAspTyrLeuSerValValLeuAsnGlnLeuCysValLeuHis450455460GluLysThrProValSerAspArgValThrLysCysCysThrGluSer465470475480LeuValAsnArgArgProCysPheSerAlaLeuGluValAspGluThr485490495TyrValProLysGluPheAsnAlaGluThrPheThrPheHisAlaAsp500505510IleCysThrLeuSerGluLysGluArgGlnIleLysLysGlnThrAla515520525LeuValGluLeuValLysHisLysProLysAlaThrLysGluGlnLeu530535540LysAlaValMetAspAspPheAlaAlaPheValGluLysCysCysLys545550555560AlaAspAspLysGluThrCysPheAlaGluGluGlyLysLysLeuVal565570575AlaAlaSerGlnAlaAlaLeuGlyLeuGlyGlyGlyGlySerGlyGly580585590GlyGlySerHisProIleProAspSerSerProLeuLeuGlnPheGly595600605GlyGlnValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThr610615620GluAlaHisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAla625630635640AspGlnSerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGly645650655ValIleGlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArg660665670ProAspGlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCys675680685SerPheArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrGlnSer690695700GluAlaHisGlyLeuProLeuHisLeuGlySerGlyArgGlyProAla705710715720ArgPheLeuProLeuProGlyLeuProProAlaProProGluProPro725730735GlyIleLeuAlaProGlnProProAspValGlySerSerAspProLeu740745750SerMetValGluProSerGlnGluArgSerProSerTyrAlaSer755760765428767PRT人工合成序列428AspAlaHisLysSerGluValAlaHisArgPheLysAspLeuGlyGlu151015GluAsnPheLysAlaLeuValLeuIleAlaPheAlaGlnTyrLeuGln202530GlnAlaProPheGluAspHisValLysLeuValAsnGluValThrGlu354045PheAlaLysThrCysValAlaAspGluSerAlaGluAsnCysAspLys505560SerLeuHisThrLeuPheGlyAspLysLeuCysThrValAlaThrLeu65707580ArgGluThrTyrGlyGluMetAlaAspCysCysAlaLysGlnGluPro859095GluArgAsnGluCysPheLeuGlnHisLysAspAspAsnProAsnLeu100105110ProArgLeuValArgProGluValAspValMetCysThrAlaPheHis115120125AspAsnGluGluThrPheLeuLysLysTyrLeuTyrGluIleAlaArg130135140ArgHisProTyrPheTyrAlaProGluLeuLeuPhePheAlaLysArg145150155160TyrLysAlaAlaPheThrGluCysCysGlnAlaAlaAspLysAlaAla165170175CysLeuLeuProLysLeuAspGluLeuArgAspGluGlyLysAlaSer180185190SerAlaLysGlnArgLeuLysCysAlaSerLeuGlnLysPheGlyGlu195200205ArgAlaPheLysAlaTrpAlaValAlaArgLeuSerGlnArgPhePro210215220LysAlaGluPheAlaGluValSerLysLeuValThrAspLeuThrLys225230235240ValHisThrGluCysCysHisGlyAspLeuLeuGluCysAlaAspAsp245250255ArgAlaAspLeuAlaLysTyrIleCysGluAsnGlnAspSerIleSer260265270SerLysLeuLysGluCysCysGluLysProLeuLeuGluLysSerHis275280285CysIleAlaGluValGluAsnAspGluMetProAlaAspLeuProSer290295300LeuAlaAlaAspPheValGluSerLysAspValCysLysAsnTyrAla305310315320GluAlaLysAspValPheLeuGlyMetPheLeuTyrGluTyrAlaArg325330335ArgHisProAspTyrSerValValLeuLeuLeuArgLeuAlaLysThr340345350TyrGluThrThrLeuGluLysCysCysAlaAlaAlaAspProHisGlu355360365CysTyrAlaLysValPheAspGluPheLysProLeuValGluGluPro370375380GlnAsnLeuIleLysGlnAsnCysGluLeuPheGluGlnLeuGlyGlu385390395400TyrLysPheGlnAsnAlaLeuLeuValArgTyrThrLysLysValPro405410415GlnValSerThrProThrLeuValGluValSerArgAsnLeuGlyLys420425430ValGlySerLysCysCysLysHisProGluAlaLysArgMetProCys435440445AlaGluAspTyrLeuSerValValLeuAsnGlnLeuCysValLeuHis450455460GluLysThrProValSerAspArgValThrLysCysCysThrGluSer465470475480LeuValAsnArgArgProCysPheSerAlaLeuGluValAspGluThr485490495TyrValProLysGluPheAsnAlaGluThrPheThrPheHisAlaAsp500505510IleCysThrLeuSerGluLysGluArgGlnIleLysLysGlnThrAla515520525LeuValGluLeuValLysHisLysProLysAlaThrLysGluGlnLeu530535540LysAlaValMetAspAspPheAlaAlaPheValGluLysCysCysLys545550555560AlaAspAspLysGluThrCysPheAlaGluGluGlyLysLysLeuVal565570575AlaAlaSerGlnAlaAlaLeuGlyLeuGlyGluThrGlySerSerGly580585590GluGlyThrHisProIleProAspSerSerProLeuLeuGlnPheGly595600605GlyGlnValArgGlnArgTyrLeuTyrThrAspAspAlaGlnGlnThr610615620GluAlaHisLeuGluIleArgGluAspGlyThrValGlyGlyAlaAla625630635640AspGlnSerProGluSerLeuLeuGlnLeuLysAlaLeuLysProGly645650655ValIleGlnIleLeuGlyValLysThrSerArgPheLeuCysGlnArg660665670ProAspGlyAlaLeuTyrGlySerLeuHisPheAspProGluAlaCys675680685SerPheArgGluLeuLeuLeuGluAspGlyTyrAsnValTyrGlnSer690695700GluAlaHisGlyLeuProLeuHisLeuGlySerGlyArgGlyProAla705710715720ArgPheLeuProLeuProGlyLeuProProAlaProProGluProPro725730735GlyIleLeuAlaProGlnProProAspValGlySerSerAspProLeu740745750SerMetValGluProSerGlnGluArgSerProSerTyrAlaSer755760765429772PRT人工合成序列429AspAlaHisLysSerGluValAlaHisArgPheLysAspLeuGlyGlu151015GluAsnPheLysAlaLeuValLeuIleAlaPheAlaGlnTyrLeuGln202530GlnAlaProPheGluAspHisValLysLeuValAsnGluValThrGlu354045PheAlaLysThrCysValAlaAspGluSerAlaGluAsnCysAspLys505560SerLeuHisThrLeuPheGlyAspLysLeuCysThrValAlaThrLeu65707580ArgGluThrTyrGlyGluMetAlaAspCysCysAlaLysGlnGluPro859095GluArgAsnGluCysPheLeuGlnHisLysAspAspAsnProAsnLeu100105110ProArgLeuValArgProGluValAspValMetCysThrAlaPheHis115120125AspAsnGluGluThrPheLeuLysLysTyrLeuTyrGluIleAlaArg130135140ArgHisProTyrPheTyrAlaProGluLeuLeuPhePheAlaLysArg145150155160TyrLysAlaAlaPheThrGluCysCysGlnAlaAlaAspLysAlaAla165170175CysLeuLeuProLysLeuAspGluLeuArgAspGluGlyLysAlaSer180185190SerAlaLysGlnArgLeuLysCysAlaSerLeuGlnLysPheGlyGlu195200205ArgAlaPheLysAlaTrpAlaValAlaArgLeuSerGlnArgPhePro210215220LysAlaGluPheAlaGluValSerLysLeuValThrAspLeuThrLys225230235240ValHisThrGluCysCysHisGlyAspLeuLeuGluCysAlaAspAsp245250255ArgAlaAspLeuAlaLysTyrIleCysGluAsnGlnAspSerIleSer260265270SerLysLeuLysGluCysCysGluLysProLeuLeuGluLysSerHis275280285CysIleAlaGluValGluAsnAspGluMetProAlaAspLeuProSer290295300LeuAlaAlaAspPheValGluSerLysAspValCysLysAsnTyrAla305310315320GluAlaLysAspValPheLeuGlyMetPheLeuTyrGluTyrAlaArg325330335ArgHisProAspTyrSerValValLeuLeuLeuArgLeuAlaLysThr340345350TyrGluThrThrLeuGluLysCysCysAlaAlaAlaAspProHisGlu355360365CysTyrAlaLysValPheAspGluPheLysProLeuValGluGluPro370375380GlnAsnLeuIleLysGlnAsnCysGluLeuPheGluGlnLeuGlyGlu385390395400TyrLysPheGlnAsnAlaLeuLeuValArgTyrThrLysLysValPro405410415GlnValSerThrProThrLeuValGluValSerArgAsnLeuGlyLys420425430ValGlySerLysCysCysLysHisProGluAlaLysArgMetProCys435440445AlaGluAspTyrLeuSerValValLeuAsnGlnLeuCysValLeuHis450455460GluLysThrProValSerAspArgValThrLysCysCysThrGluSer465470475480LeuValAsnArgArgProCysPheSerAlaLeuGluValAspGluThr485490495TyrValProLysGluPheAsnAlaGluThrPheThrPheHisAlaAsp500505510IleCysThrLeuSerGluLysGluArgGlnIleLysLysGlnThrAla515520525LeuValGluLeuValLysHisLysProLysAlaThrLysGluGlnLeu530535540LysAlaValMetAspAspPheAlaAlaPheValGluLysCysCysLys545550555560AlaAspAspLysGluThrCysPheAlaGluGluGlyLysLysLeuVal565570575AlaAlaSerGlnAlaAlaLeuGlyLeuGlyGlyGlyGlySerGlyGly580585590GlyGlySerGlyGlyGlyGlySerHisProIleProAspSerSerPro595600605LeuLeuGlnPheGlyGlyGlnValArgGlnArgTyrLeuTyrThrAsp610615620AspAlaGlnGlnThrGluAlaHisLeuGluIleArgGluAspGlyThr625630635640ValGlyGlyAlaAlaAspGlnSerProGluSerLeuLeuGlnLeuLys645650655AlaLeuLysProGlyValIleGlnIleLeuGlyValLysThrSerArg660665670PheLeuCysGlnArgProAspGlyAlaLeuTyrGlySerLeuHisPhe675680685AspProGluAlaCysSerPheArgGluLeuLeuLeuGluAspGlyTyr690695700AsnValTyrGlnSerGluAlaHisGlyLeuProLeuHisLeuGlySer705710715720GlyArgGlyProAlaArgPheLeuProLeuProGlyLeuProProAla725730735ProProGluProProGlyIleLeuAlaProGlnProProAspValGly740745750SerSerAspProLeuSerMetValGluProSerGlnGluArgSerPro755760765SerTyrAlaSer770430772PRT人工合成序列430AspAlaHisLysSerGluValAlaHisArgPheLysAspLeuGlyGlu151015GluAsnPheLysAlaLeuValLeuIleAlaPheAlaGlnTyrLeuGln202530GlnAlaProPheGluAspHisValLysLeuValAsnGluValThrGlu354045PheAlaLysThrCysValAlaAspGluSerAlaGluAsnCysAspLys505560SerLeuHisThrLeuPheGlyAspLysLeuCysThrValAlaThrLeu65707580ArgGluThrTyrGlyGluMetAlaAspCysCysAlaLysGlnGluPro859095GluArgAsnGluCysPheLeuGlnHisLysAspAspAsnProAsnLeu100105110ProArgLeuValArgProGluValAspValMetCysThrAlaPheHis115120125AspAsnGluGluThrPheLeuLysLysTyrLeuTyrGluIleAlaArg130135140ArgHisProTyrPheTyrAlaProGluLeuLeuPhePheAlaLysArg145150155160TyrLysAlaAlaPheThrGluCysCysGlnAlaAlaAspLysAlaAla165170175CysLeuLeuProLysLeuAspGluLeuArgAspGluGlyLysAlaSer180185190SerAlaLysGlnArgLeuLysCysAlaSerLeuGlnLysPheGlyGlu195200205ArgAlaPheLysAlaTrpAlaValAlaArgLeuSerGlnArgPhePro210215220LysAlaGluPheAlaGluValSerLysLeuValThrAspLeuThrLys225230235240ValHisThrGluCysCysHisGlyAspLeuLeuGluCysAlaAspAsp245250255ArgAlaAspLeuAlaLysTyrIleCysGluAsnGlnAspSerIleSer260265270SerLysLeuLysGluCysCysGluLysProLeuLeuGluLysSerHis275280285CysIleAlaGluValGluAsnAspGluMetProAlaAspLeuProSer290295300LeuAlaAlaAspPheValGluSerLysAspValCysLysAsnTyrAla305310315320GluAlaLysAspValPheLeuGlyMetPheLeuTyrGluTyrAlaArg325330335ArgHisProAspTyrSerValValLeuLeuLeuArgLeuAlaLysThr340345350TyrGluThrThrLeuGluLysCysCysAlaAlaAlaAspProHisGlu355360365CysTyrAlaLysValPheAspGluPheLysProLeuValGluGluPro370375380GlnAsnLeuIleLysGlnAsnCysGluLeuPheGluGlnLeuGlyGlu385390395400TyrLysPheGlnAsnAlaLeuLeuValArgTyrThrLysLysValPro405410415GlnValSerThrProThrLeuValGluValSerArgAsnLeuGlyLys420425430ValGlySerLysCysCysLysHisProGluAlaLysArgMetProCys435440445AlaGluAspTyrLeuSerValValLeuAsnGlnLeuCysValLeuHis450455460GluLysThrProValSerAspArgValThrLysCysCysThrGluSer465470475480LeuValAsnArgArgProCysPheSerAlaLeuGluValAspGluThr485490495TyrValProLysGluPheAsnAlaGluThrPheThrPheHisAlaAsp500505510IleCysThrLeuSerGluLysGluArgGlnIleLysLysGlnThrAla515520525LeuValGluLeuValLysHisLysProLysAlaThrLysGluGlnLeu530535540LysAlaValMetAspAspPheAlaAlaPheValGluLysCysCysLys545550555560AlaAspAspLysGluThrCysPheAlaGluGluGlyLysLysLeuVal565570575AlaAlaSerGlnAlaAlaLeuGlyLeuGlyGluThrGlySerSerGly580585590GluGlyThrGlySerThrGlySerHisProIleProAspSerSerPro595600605LeuLeuGlnPheGlyGlyGlnValArgGlnArgTyrLeuTyrThrAsp610615620AspAlaGlnGlnThrGluAlaHisLeuGluIleArgGluAspGlyThr625630635640ValGlyGlyAlaAlaAspGlnSerProGluSerLeuLeuGlnLeuLys645650655AlaLeuLysProGlyValIleGlnIleLeuGlyValLysThrSerArg660665670PheLeuCysGlnArgProAspGlyAlaLeuTyrGlySerLeuHisPhe675680685AspProGluAlaCysSerPheArgGluLeuLeuLeuGluAspGlyTyr690695700AsnValTyrGlnSerGluAlaHisGlyLeuProLeuHisLeuGlySer705710715720GlyArgGlyProAlaArgPheLeuProLeuProGlyLeuProProAla725730735ProProGluProProGlyIleLeuAlaProGlnProProAspValGly740745750SerSerAspProLeuSerMetValGluProSerGlnGluArgSerPro755760765SerTyrAlaSer770431777PRT人工合成序列431AspAlaHisLysSerGluValAlaHisArgPheLysAspLeuGlyGlu151015GluAsnPheLysAlaLeuValLeuIleAlaPheAlaGlnTyrLeuGln202530GlnAlaProPheGluAspHisValLysLeuValAsnGluValThrGlu354045PheAlaLysThrCysValAlaAspGluSerAlaGluAsnCysAspLys505560SerLeuHisThrLeuPheGlyAspLysLeuCysThrValAlaThrLeu65707580ArgGluThrTyrGlyGluMetAlaAspCysCysAlaLysGlnGluPro859095GluArgAsnGluCysPheLeuGlnHisLysAspAspAsnProAsnLeu100105110ProArgLeuValArgProGluValAspValMetCysThrAlaPheHis115120125AspAsnGluGluThrPheLeuLysLysTyrLeuTyrGluIleAlaArg130135140ArgHisProTyrPheTyrAlaProGluLeuLeuPhePheAlaLysArg145150155160TyrLysAlaAlaPheThrGluCysCysGlnAlaAlaAspLysAlaAla165170175CysLeuLeuProLysLeuAspGluLeuArgAspGluGlyLysAlaSer180185190SerAlaLysGlnArgLeuLysCysAlaSerLeuGlnLysPheGlyGlu195200205ArgAlaPheLysAlaTrpAlaValAlaArgLeuSerGlnArgPhePro210215220LysAlaGluPheAlaGluValSerLysLeuValThrAspLeuThrLys225230235240ValHisThrGluCysCysHisGlyAspLeuLeuGluCysAlaAspAsp245250255ArgAlaAspLeuAlaLysTyrIleCysGluAsnGlnAspSerIleSer260265270SerLysLeuLysGluCysCysGluLysProLeuLeuGluLysSerHis275280285CysIleAlaGluValGluAsnAspGluMetProAlaAspLeuProSer290295300LeuAlaAlaAspPheValGluSerLysAspValCysLysAsnTyrAla305310315320GluAlaLysAspValPheLeuGlyMetPheLeuTyrGluTyrAlaArg325330335ArgHisProAspTyrSerValValLeuLeuLeuArgLeuAlaLysThr340345350TyrGluThrThrLeuGluLysCysCysAlaAlaAlaAspProHisGlu355360365CysTyrAlaLysValPheAspGluPheLysProLeuValGluGluPro370375380GlnAsnLeuIleLysGlnAsnCysGluLeuPheGluGlnLeuGlyGlu385390395400TyrLysPheGlnAsnAlaLeuLeuValArgTyrThrLysLysValPro405410415GlnValSerThrProThrLeuValGluValSerArgAsnLeuGlyLys420425430ValGlySerLysCysCysLysHisProGluAlaLysArgMetProCys435440445AlaGluAspTyrLeuSerValValLeuAsnGlnLeuCysValLeuHis450455460GluLysThrProValSerAspArgValThrLysCysCysThrGluSer465470475480LeuValAsnArgArgProCysPheSerAlaLeuGluValAspGluThr485490495TyrValProLysGluPheAsnAlaGluThrPheThrPheHisAlaAsp500505510IleCysThrLeuSerGluLysGluArgGlnIleLysLysGlnThrAla515520525LeuValGluLeuValLysHisLysProLysAlaThrLysGluGlnLeu530535540LysAlaValMetAspAspPheAlaAlaPheValGluLysCysCysLys545550555560AlaAspAspLysGluThrCysPheAlaGluGluGlyLysLysLeuVal565570575AlaAlaSerGlnAlaAlaLeuGlyLeuGlyGlyGlyGlySerGlyGly580585590GlyGlySerGlyGlyGlyGlySerGlyGlyGlyGlySerHisProIle595600605ProAspSerSerProLeuLeuGlnPheGlyGlyGlnValArgGlnArg610615620TyrLeuTyrThrAspAspAlaGlnGlnThrGluAlaHisLeuGluIle625630635640ArgGluAspGlyThrValGlyGlyAlaAlaAspGlnSerProGluSer645650655LeuLeuGlnLeuLysAlaLeuLysProGlyValIleGlnIleLeuGly660665670ValLysThrSerArgPheLeuCysGlnArgProAspGlyAlaLeuTyr675680685GlySerLeuHisPheAspProGluAlaCysSerPheArgGluLeuLeu690695700LeuGluAspGlyTyrAsnValTyrGlnSerGluAlaHisGlyLeuPro705710715720LeuHisLeuGlySerGlyArgGlyProAlaArgPheLeuProLeuPro725730735GlyLeuProProAlaProProGluProProGlyIleLe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AlaLeuGlyGlyGlyGlyGlySerGlyGlyGly580585590GlySerGlyGlyGlyGlySerHisProIleProAspSerSerProLeu595600605LeuGlnPheGlyGlyGlnValArgGlnArgTyrLeuTyrThrAspAsp610615620AlaGlnGlnThrGluAlaHisLeuGluIleArgGluAspGlyThrVal625630635640GlyGlyAlaAlaAspGlnSerProGluSerLeuLeuGlnLeuLysAla645650655LeuLysProGlyValIleGlnIleLeuGlyValLysThrSerArgPhe660665670LeuCysGlnArgProAspGlyAlaLeuTyrGlySerLeuHisPheAsp675680685ProGluAlaCysSerPheArgGluLeuLeuLeuGluAspGlyTyrAsn690695700ValTyrGlnSerGluAlaHisGlyLeuProLeuHisLeuGlySerGly705710715720ArgGlyProAlaArgPheLeuProLeuProGlyLeuProProAlaPro725730735ProGluProProGlyIleLeuAlaProGlnProProAspValGlySer740745750SerAspProLeuSerMetValGluProSerGlnGluArgSerProSer755760765TyrAlaSer770435771PRT人工合成序列435AspAlaHisLysSerGluValAlaHisArgPheLysAspLeuGlyGlu151015GluAsnPheLysAlaLeuValLeuIleAlaPheAlaGlnTyrLeuGln202530GlnAlaProPheGluAspHisValLysLeuValAsnGluValThrGlu354045PheAlaLysThrCysValAlaAspGluSerAlaGluAsnCysAspLys505560SerLeuHisThrLeuPheGlyAspLysLeuCysThrValAlaThrLeu65707580ArgGluThrTyrGlyGluMetAlaAspCysCysAlaLysGlnGluPro859095GluArgAsnGluCysPheLeuGlnHisLysAspAspAsnProAsnLeu100105110ProArgLeuValArgProGluValAspValMetCysThrAlaPheHis115120125AspAsnGluGluThrPheLeuLysLysTyrLeuTyrGluIleAlaArg130135140ArgHisProTyrPheTyrAlaProGluLeuLeuPhePheAlaLysArg145150155160TyrLysAlaAlaPheThrGluCysCysGlnAlaAlaAspLysAlaAla165170175CysLeuLeuProLysLeuAspGluLeuArgAspGluGlyLysAlaSer180185190SerAlaLysGlnArgLeuLysCysAlaSerLeuGlnLysPheGlyGlu195200205ArgAlaPheLysAlaTrpAlaValAlaArgLeuSerGlnArgPhePro210215220LysAlaGluPheAlaGluValSerLysLeuValThrAspLeuThrLys225230235240ValHisThrGluCysCysHisGlyAspLeuLeuGluCysAlaAspAsp245250255ArgAlaAspLeuAlaLysTyrIleCysGluAsnGlnAspSerIleSer260265270SerLysLeuLysGluCysCysGluLysProLeuLeuGluLysSerHis275280285CysIleAlaGluValGluAsnAspGluMetProAlaAspLeuProSer290295300LeuAlaAlaAspPheValGluSerLysAspValCysLysAsnTyrAla305310315320GluAlaLysAspValPheLeuGlyMetPheLeuTyrGluTyrAlaArg325330335ArgHisProAspTyrSerValValLeuLeuLeuArgLeuAlaLysThr340345350TyrGluThrThrLeuGluLysCysCysAlaAlaAlaAspProHisGlu355360365CysTyrAlaLysValPheAspGluPheLysProLeuValGluGluPro370375380GlnAsnLeuIleLysGlnAsnCysGluLeuPheGluGlnLeuGlyGlu385390395400TyrLysPheGlnAsnAlaLeuLeuValArgTyrThrLysLysValPro405410415GlnValSerThrProThrLeuValGluValSerArgAsnLeuGlyLys420425430ValGlySerLysCysCysLysHisProGluAlaLysArgMetProCys435440445AlaGluAspTyrLeuSerValValLeuAsnGlnLeuCysValLeuHis450455460GluLysThrProValSerAspArgValThrLysCysCysThrGluSer465470475480LeuValAsnArgArgProCysPheSerAlaLeuGluValAspGluThr485490495TyrValProLysGluPheAsnAlaGluThrPheThrPheHisAlaAsp500505510IleCysThrLeuSerGluLysGluArgGlnIleLysLysGlnThrAla515520525LeuValGluLeuValLysHisLysProLysAlaThrLysGluGlnLeu530535540LysAlaValMetAspAspPheAlaAlaPheValGluLysCysCysLys545550555560AlaAspAspLysGluThrCysPheAlaGluGluGlyLysLysLeuVal565570575AlaAlaSerGlnAlaAlaLeuGlyGlyGluThrGlySerSerGlyGlu580585590GlyThrGlySerThrGlySerHisProIleProAspSerSerProLeu595600605LeuGlnPheGlyGlyGlnValArgGlnArgTyrLeuTyrThrAspAsp610615620AlaGlnGlnThrGluAlaHisLeuGluIleArgGluAspGlyThrVal625630635640GlyGlyAlaAlaAspGlnSerProGluSerLeuLeuGlnLeuLysAla645650655LeuLysProGlyValIleGlnIleLeuGlyValLysThrSerArgPhe660665670LeuCysGlnArgProAspGlyAlaLeuTyrGlySerLeuHisPheAsp675680685ProGluAlaCysSerPheArgGluLeuLeuLeuGluAspGlyTyrAsn690695700ValTyrGlnSerGluAlaHisGlyLeuProLeuHisLeuGlySerGly705710715720ArgGlyProAlaArgPheLeuProLeuProGlyLeuProProAlaPro725730735ProGluProProGlyIleLeuAlaProGlnProProAspValGlySer740745750SerAspProLeuSerMetValGluProSerGlnGluArgSerProSer755760765TyrAlaSer770436780PRT人工合成序列436AspAlaHisLysSerGluValAlaHisArgPheLysAspLeuGlyGlu151015GluAsnPheLysAlaLeuValLeuIleAlaPheAlaGlnTyrLeuGln202530GlnAlaProPheGluAspHisValLysLeuValAsnGluValThrGlu354045PheAlaLysThrCysValAlaAspGluSerAlaGluAsnCysAspLys505560SerLeuHisThrLeuPheGlyAspLysLeuCysThrValAlaThrLeu65707580ArgGluThrTyrGlyGluMetAlaAspCysCysAlaLysGlnGluPro859095GluArgAsnGluCysPheLeuGlnHisLysAspAspAsnProAsnLeu100105110ProArgLeuValArgProGluValAspValMetCysThrAlaPheHis115120125AspAsnGluGluThrPheLeuLysLysTyrLeuTyrGluIleAlaArg130135140ArgHisProTyrPheTyrAlaProGluLeuLeuPhePheAlaLysArg145150155160TyrLysAlaAlaPheThrGluCysCysGlnAlaAlaAspLysAlaAla165170175CysLeuLeuProLysLeuAspGluLeuArgAspGluGlyLysAlaSer180185190SerAlaLysGlnArgLeuLysCysAlaSerLeuGlnLysPheGlyGlu195200205ArgAlaPheLysAlaTrpAlaValAlaArgLeuSerGlnArgPhePro210215220LysAlaGluPheAlaGluValSerLysLeuValThrAspLeuThrLys225230235240ValHisThrGluCysCysHisGlyAspLeuLeuGluCysAlaAspAsp245250255ArgAlaAspLeuAlaLysTyrIleCysGluAsnGlnAspSerIleSer260265270SerLysLeuLysGluCysCysGluLysProLeuLeuGluLysSerHis275280285CysIleAlaGluValGluAsnAspGluMetProAlaAspLeuProSer290295300LeuAlaAlaAspPheValGluSerLysAspValCysLysAsnTyrAla305310315320GluAlaLysAspValPheLeuGlyMetPheLeuTyrGluTyrAlaArg325330335ArgHisProAspTyrSerValValLeuLeuLeuArgLeuAlaLysThr340345350TyrGluThrThrLeuGluLysCysCysAlaAlaAlaAspProHisGlu355360365CysTyrAlaLysValPheAspGluPheLysProLeuValGluGluPro370375380GlnAsnLeuIleLysGlnAsnCysGluLeuPheGluGlnLeuGlyGlu385390395400TyrLysPheGlnAsnAlaLeuLeuValArgTyrThrLysLysValPro405410415GlnValSerThrProThrLeuValGluValSerArgAsnLeuG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权利要求：1.修饰的FGF-21多肽在制备用于治疗纤维化相关疾病的药物中的用途，所述纤维化相关疾病选自非酒精性脂肪性肝炎NASH、肝纤维化或肝硬化，其中所述修饰的FGF-21多肽包含SEQIDNO:201的多肽，其中其对-乙酰基-苯丙氨酸残基连接至聚乙二醇部分。

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