CN111741975A - 抗血管生成素-2抗体及其用途 - Google Patents
抗血管生成素-2抗体及其用途 Download PDFInfo
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- CN111741975A CN111741975A CN201980014282.1A CN201980014282A CN111741975A CN 111741975 A CN111741975 A CN 111741975A CN 201980014282 A CN201980014282 A CN 201980014282A CN 111741975 A CN111741975 A CN 111741975A
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Abstract
本发明涉及与血管生成素‑2(Ang2)结合的小鼠抗体、由其衍生的人源化抗Ang2抗体以及它们的用途。所述抗Ang2抗体具有激活所述Tie2受体以及中和Ang2的双重功能。所述抗Ang2抗体显示出使异常和病理性血管正常化的特性,并且因此针对与异常血管相关的各种疾病和障碍展现出治疗功效。本发明提供了血管生成抑制剂和包含所述抗体作为有效成分的用于预防和治疗与异常Ang2表达和Tie2调节异常相关的疾病的组合物,以及包含所述抗体的用于诊断与Ang2抑制和Tie2激活相关的疾病的组合物。
Description
技术领域
本发明包括抗Ang2抗体或其抗原结合片段,所述抗Ang2抗体或其抗原结合片段与被称为控制血管形成和维持的配体的血管生成素-2(Ang2)特异性地结合;含有所述抗体或其抗原结合片段的药物组合物;编码所述抗体或其抗原结合片段的核酸;包含所述核酸的载体;用所述载体转化的宿主细胞;以及用于产生所述抗体或其抗原结合片段的方法。
背景技术
在生物体的发育、生长、维持和体内平衡过程中,血管生成是通过多种调节因子动态发生的。在此过程中新形成的血管充当各种生物材料(如周围细胞中的营养素、氧气和激素)的运输通道。功能和结构异常的血管是各种疾病的发生和发展的直接或间接原因。肿瘤血管由于其有缺陷的功能和结构而加重了缺氧,从而导致肿瘤进展和转移至其他组织,并且还导致抗癌药物向肿瘤块核心的递送不良。除了癌症之外,在其他各种疾病和病症中也发现了有缺陷的血管。其例子包括各种眼病(例如,糖尿病性黄斑水肿、湿性年龄相关性黄斑变性)、病毒感染和急性炎症反应(如败血症)。因此,如果可获得能够使病理血管正常化的治疗剂,则可以将其应用于治疗患有血管异常的各种患者。
血管生成素家族在血管的形成和维持中起着重要作用,并且由四种血管生成素(Ang1、Ang2、Ang3和Ang4)组成。血管生成素-1(Ang1)与存在于血管内皮细胞表面上的Tie2受体结合,以磷酸化并激活Tie2受体,从而导致血管稳定。另一方面,血管生成素-2(Ang2)与Tie2受体结合,但充当拮抗剂以诱导Tie2受体的失活,从而导致血管不稳定和血管渗漏。据报道,在癌症患者、眼病、病毒和细菌感染以及炎性疾病的血液中,Ang2的表达水平大大提高(Saharinen P等人,2017,Nature Review Drug Discovery)。然而,还已知Ang2在若干个过程(包括淋巴管形成和维持)中充当激动剂以诱导Tie2受体的激活,并且因此认为Ang2根据情境执行多种功能。
Ang2结合抗体已在若干个文献(例如,US 7,658,924和US 8,987,420)中报道。已知迄今为止报道的大多数Ang2抗体抑制Ang2与Tie2的结合,并因此通过这种Ang2中和功效抑制新血管的形成。当前,多种Ang2抗体正在各种癌症患者中进行临床测试,但其抗癌功效被认为是不足的。例如,由Amgen进行的3期临床试验显示,所述Ang2抗体在卵巢癌患者中的抗癌功效是不显著的(Marth C等人,2017,Eur.J.Cancer)。
除了抗体之外,还已报道了与Tie2受体直接结合以诱导Tie2磷酸化和激活的重组蛋白。其例子包括由五个血管生成素-1蛋白片段组成的COMP-Angl(Cho等人,2004,PNAS)和Vasculotide(David S等人,2011,Am J Physiol Lung Cell Mol Physiol)肽。然而,人们认为这些蛋白质具有非常短的半衰期和不稳定的物理化学特性。此外,存在一种称为VE-PTP的磷酸酶,其从磷酸化Tie2去除磷酸基团以使Tie2失活,并且还开发了一种低分子化合物(AKB-9778),其通过抑制酶VE-PTP的活性间接维持Tie2活性(Goel S,2013,J NatlCancer Inst)。然而,此化合物的缺点是除了Tie2之外还激活其他受体(Frye M,2015,JExp.Med,Hayashi M,2013,Nature Communication,Mellberg S等人,2009,FASEB J.)。
发明内容
本发明涉及特异性地结合人血管生成素-2并诱导Tie2激活的抗体或其抗原结合片段,其中所述抗体或其抗原结合片段与SEQ ID NO:1的氨基酸289-299、SEQ ID NO:1的氨基酸316-322或SEQ ID NO:1的氨基酸336-353结合,如通过氢/氘交换方法所确定。
所述抗体或其抗原结合片段可以与人和小鼠Ang2结合。所述抗体可以是多克隆的或单克隆的。所述抗原结合片段可以是scFv或Fab。所述抗体或其片段可以是人源化的。
在另一方面,本发明涉及包含以下各项的抗体或抗原结合片段:
(a)具有SEQ ID NO:3的HCDR1氨基酸序列、SEQ ID NO:4的HCDR2氨基酸序列和SEQID NO:5的HCDR3氨基酸序列的重链可变区的互补决定区(CDR);和
(b)含有SEQ ID NO:6的LCDR1氨基酸序列、SEQ ID NO:7的LCDR2氨基酸序列和SEQID NO:8的LCDR3氨基酸序列的轻链可变区的CDR。
在另一方面,本发明涉及包含以下各项的抗体或抗原结合片段:
(a)具有SEQ ID NO:13的HCDR1氨基酸序列、SEQ ID NO:14的HCDR2氨基酸序列和SEQ ID NO:15的HCDR3氨基酸序列的重链可变区的互补决定区(CDR);和
(b)含有SEQ ID NO:16的LCDR1氨基酸序列、SEQ ID NO:17的LCDR2氨基酸序列和SEQ ID NO:18的LCDR3氨基酸序列的轻链可变区的CDR。
在一方面,本发明涉及抗体或其抗原结合片段,所述抗体或其抗原结合片段包含由以保藏号KCLRF-BP-00417或KCLRF-BP-00418保藏的细胞系产生的抗体的互补决定区(CDR)。
在又另一方面,本发明涉及药物组合物,其包含与药学上可接受的载体相关联的药学有效量的上述抗体或其抗原结合片段。所述药物组合物可以进一步包含在化学疗法中使用的小分子抑制剂或血管内皮生长因子(VEGF)拮抗剂。在一方面,所述VEGF拮抗剂可以是抗VEGF抗体、VEGF抑制性融合蛋白或小分子激酶抑制剂。
在又另一方面,本发明涉及用于抑制患者中肿瘤生长的方法,其包括向所述患者给予包含上述抗体或抗原结合片段的药物组合物。所述方法可以进一步包括与本发明抗体或其片段的给予同时或逐步给予在化学疗法中使用的小分子抑制剂或血管内皮生长因子(VEGF)拮抗剂。所述VEGF拮抗剂可以是抗VEGF抗体、VEGF抑制性融合蛋白或小分子激酶抑制剂。
在又另一方面,本发明涉及用于在眼病患者中抑制脉络膜新血管形成、抑制眼血管渗漏或同时触发脉络膜毛细血管再生的方法,所述方法包括向所述患者给予上述药物组合物。所述方法可以进一步包括与本发明抗体或其片段的给予同时或逐步给予在化学疗法中使用的小分子抑制剂或血管内皮生长因子(VEGF)拮抗剂。所述VEGF拮抗剂可以是抗VEGF抗体、VEGF抑制性融合蛋白或小分子激酶抑制剂。所述眼病是湿性年龄相关性黄斑变性(wAMD)、糖尿病性黄斑水肿(DME)或糖尿病性视网膜病(DR)。
本发明的这些和其他目的将从本发明的以下描述、本发明所附的附图和所附的权利要求书中得到更充分的理解。
附图说明
从下文给出的详细描述和附图中将更全面地理解本发明,这些详细描述和附图仅以说明的方式给出并且因此不限制本发明,并且其中;
图1.由抗Ang2抗体诱导的Akt磷酸化。将HUVEC进行血清饥饿6hr,并且在不存在或存在人Ang2(1μg/ml)的情况下,将其与COMP-Ang1(CA1,0.5μg/ml)或抗Ang2抗体(分别为对照、2C8、4B9、2F10和4E2)一起孵育30min。对细胞裂解物进行SDS-PAGE/蛋白质印记,并用抗磷酸Akt(S473)或抗Akt抗体探测印迹。
图2.示意图示出抗Ang2抗体的表位,已通过氢/氘交换质谱法对其进行了分析。用氘标记单独的重组hAng2-RBD或hAng2-RBD/Ang2-抗体复合物。在胃蛋白酶柱中消化标记的蛋白,并通过质谱法对其进行分析。分析了单独的hAng2-RBD和hAng2-RBD/Ang-2抗体复合物的氘摄取,并比较了氘摄取的差异。氘摄取的质量差异超过0.5-1Da的肽被确定为介导与抗Ang2抗体结合的特异性表位。2C8表位(红色)和4B9表位(绿色)在使用PyMol软件生成的Ang2-RBD晶体结构(PDB:2GY7)的图像中被可视化。
图3.通过人源化抗Ang2抗体4B9H11和2C8H11进行的剂量依赖性Akt磷酸化(pAkt)。将血清饥饿的HUVEC与人Ang2、抗Ang2抗体或人Ang2以及各种浓度的抗Ang2抗体一起孵育30min。对细胞裂解物进行SDS-PAGE/蛋白质印迹。
图4.通过人源化抗Ang2抗体4B9H11和2C8H11进行的剂量依赖性Tie2磷酸化(pTie2)。通过免疫沉淀和蛋白质印迹(Western)分析研究了4B9H11(图4A)和2C8H11(图4B)抗体诱导Tie2磷酸化的能力。将血清饥饿的HUVEC与人Ang2、单独的抗Ang2抗体或人Ang2以及各种浓度的抗Ang2抗体一起孵育30min。用抗Tie2抗体对细胞裂解物进行免疫沉淀,随后进行SDS-PAGE/蛋白质印迹分析。
图5.由人源化Ang2抗体引起的Tie2受体聚集和FOXO1易位。将HUVEC进行血清饥饿6hr,并且将其与COMP-Ang1(CA1)、Ang2(A2)或Ang2以及抗Ang2抗体(对照Ab、2C8H11或4B9H11)一起孵育30min。在固定后,将HUVEC用DAPI(蓝色)、抗Tie2抗体(绿色)、抗FOXO1抗体(红色)和抗人Fc(青色)染色,以研究细胞表面处的Tie2聚集、FOXO1从细胞核的易位以及人源化Ang2抗体在细胞间连接区域中的存在。箭头指示在细胞间接触处聚集的Tie2和共定位的Ang2抗体。
图6.HUVEC中的细胞间连接中,Tie2受体聚集、FOXO1易位和Ang2抗体定位的时间进程。将血清饥饿的HUVEC与抗Ang2抗体(对照Ab或2C8H11)一起孵育不同时间点(从10min至240min)。在细胞固定后,通过用抗Tie2抗体染色来研究细胞表面处聚集的Tie2受体和内吞的Tie2受体。用抗人Fc抗体探测细胞表面和胞质溶胶处的人源化抗Ang2抗体。箭头指示在细胞间接触处聚集的Tie2和共定位的Ang2抗体。
图7.通过人源化抗Ang2抗体抑制血管通透性。将HUVEC接种在穿透(transwell)室上并生长3天。在100%汇合度时,将HUVEC用COMP-Ang1(CA1,0.5μg/ml)、Ang2(A2,1μg/ml)、Ang2以及对照Ab(A2+对照Ab,1μg/ml)、2C8H11(A2+2C8H11,1μg/ml)或4B9H11(A2+4B9H11,1μg/ml)预处理30min,并用TNF-a(100ng/ml)处理22hr进入上部室。在上部室中添加FITC-葡聚糖20min后,通过测量下部室中的FITC荧光来评估血管通透性。值是平均值±SD。通过单向ANOVA,*p<0.05,**p<0.01,***p<0.001。
图8.通过ELISA得到的抗Ang2抗体对小鼠Ang2的EC50值。通过分析用ELISA得到的EC50来测量人源化抗Ang2抗体对小鼠Ang2(mAng2)的结合亲和力。将重组mAng2包被,并与连续稀释的抗Ang2抗体4B9H11和2C8H11一起孵育。接下来,使板与抗人IgG(Fab)-HRP二抗反应。用TMB溶液处理板,并在450nm处测量吸光度。使用PerkinElmer的WorkOut 2.5程序分析EC50值。
图9.人源化2C8H11抗体和顺铂(Cpt)在LLC肿瘤模型中抑制肿瘤生长。在肿瘤植入后7天开始,在按所指示处理的小鼠中比较LLC肿瘤生长。黑色箭头指示抗体的注射,而红色箭头指示Cpt的单次注射。对于各组,n=7-9。值是平均值±SD。相对于Fc,*p<0.05;相对于Fc+Cpt,#p<0.05。
图10.通过人源化2C8H11抗体对肿瘤血管的正常化作用。在LLC皮下肿瘤模型中比较了肿瘤上的PDGFRβ+周细胞覆盖率和肿瘤内区域中的CD31+BV。比例尺,100μm。对于各组,n=5。值是平均值±SD。相对于Fc,*p<0.05;相对于Fc+Cpt,#p<0.05。
图11.通过人源化2C8H11抗体引起的在肿瘤血管中的缺氧减少灌注增加。在LLC肿瘤中分析并比较了肿瘤血管的凝集素灌注和低氧探针+缺氧面积。低氧探针+面积表示为每总截面积的百分比。比例尺,100μm。对于各组,n=5。值是平均值±SD。相对于Fc,*p<0.05;相对于Fc+Cpt,#p<0.05。
图12.通过人源化2C8H11抗体增强Cpt药物向肿瘤核心的递送。使用抗Cpt修饰的DNA抗体,在第21天收获的肿瘤中将Cpt+面积成像。将Cpt+面积测量为每总截面积的百分比。比例尺,100μm。对于各组,n=5。值是平均值±SD。相对于Fc+Cpt,#p<0.05。
图13.在激光诱导的CNV模型中通过玻璃体内注射2C8H11抗体抑制CNV消退和血管渗漏。在激光光凝后7天进行抗体的玻璃体内给予。测量CD31+CNV体积,并将CNV周围的渗漏面积计算为在激光光凝后6天和/或14天在FA图像中测量的总超荧光面积除以在ICGA图像中测量的总CNV面积。比例尺,100μm。对于各组,n=11。值是平均值±SD。通过单向ANOVA,接着是学生-纽曼-科伊尔斯事后检验,***p<0.001;通过配对的学生t检验,###p<0.001。
图14.通过玻璃体内注射2C8H11抗体引起的CNV消退和脉络膜毛细血管再生。在激光光凝后7天进行抗体的玻璃体内给予。通过在激光光凝后6、14、21和35天对眼睛进行OCTA成像,测量CNV体积(用白色虚线框划出的区域)和围绕CNV的无血管空间(用黄色虚线框划出的区域)。对于各组,n=11。值是平均值±SD。通过单向ANOVA,接着是学生-纽曼-科伊尔斯事后检验,相对于Fc,*p<0.05,**p<0.005。
图15.2C8H11抗体和CD31在CNV内皮细胞中的共定位。在激光光凝后1天进行2C8H11抗体的皮下给予。在激光光凝后2、4和8天,通过抗人IgG抗体直接检测2C8H11抗体和CD31在CNV内皮细胞中的共定位。
图16.皮下注射的2C8H11抗体的CNV抑制作用。在激光光凝后1天进行2C8H11抗体的皮下给予。在激光光凝后8天测量CD31+CNV体积。比例尺,100μm。对于各组,n=10。值是平均值±SD。通过未配对的学生t检验,***p<0.001。
具体实施方式
除非另有定义,否则本文中所用的所有技术和科学术语都具有与本公开文本所属领域技术人员所了解的相同的含义。通常,本文中所用的术语为本领域所熟知并且以普通方式使用。
在本申请中,“一个”和“一种”用于指单个和多个对象两者。
在一方面,本发明涉及特异性地结合人血管生成素-2并诱导Tie2激活的抗体或其抗原结合片段,其中所述抗体或其抗原结合片段与SEQ ID NO:115的氨基酸、SEQ ID NO:116的氨基酸或SEQ ID NO:117的氨基酸结合。
SEQ ID NO:115的氨基酸对应于SEQ ID NO:1的氨基酸336-353,SEQ ID NO:116的氨基酸对应于SEQ ID NO:1的氨基酸289-299,并且SEQ ID NO:117的氨基酸对应于SEQ IDNO:1的氨基酸316-322。
如本文所用,术语“与Ang2特异性结合的抗体”是指与Ang2结合从而导致抑制Ang2的生物活性的抗体,并且可与“抗Ang2抗体”、“Ang2结合抗体”互换使用。
本文所用的“抗体”是与特定抗原发生免疫反应的免疫球蛋白分子,并且意指充当特异性识别抗原的受体的蛋白质分子,并且可以包括多克隆抗体、单克隆抗体(单个克隆抗体)、全抗体和抗体片段中的全部。此外,所述抗体可以包括嵌合抗体(例如,人源化鼠抗体)和二价或双特异性分子(例如,双特异性抗体)、双抗体、三抗体和四抗体。
全抗体具有包含两条全长轻链和两条全长重链的结构,并且每条轻链可以通过二硫键与重链连接。全抗体包括IgA、IgD、IgE、IgM和IgG,并且IgG是亚型并包括IgG1、IgG2、IgG3和IgG4。
在本公开文本中,所述抗体或其抗原结合片段可以与人和小鼠Ang2结合。
抗体片段意指保留抗原结合功能的片段,并且包括Fab、Fab'、F(ab')2、scFv和Fv等。
Fab具有轻链和重链的可变区以及轻链的恒定区和重链的第一恒定区(CH1结构域)的结构,并且具有一个抗原结合位点。Fab'与Fab的不同之处在于,Fab′具有铰链区,所述铰链区包括重链CH1结构域的C末端处的一个或多个半胱氨酸残基。F(ab')2抗体是通过实现Fab'的铰链区中的半胱氨酸残基的二硫键产生的。
Fv(可变片段)是指仅具有重链可变区和轻链可变区的最小抗体片段。在双链Fv(dsFv)中,重链可变区和轻链可变区通过二硫键连接。在单链Fv(scFv)中,重链可变区和轻链可变区通常使用肽接头通过共价键连接。这些抗体片段可以通过使用蛋白水解酶获得(例如,Fab可以通过用木瓜蛋白酶限制性切割全抗体来获得,并且F(ab')2片段可以通过用胃蛋白酶切割来获得),并且可以通过重组DNA技术来构建(例如,通过使用编码抗体的重链或其可变区的DNA和编码轻链或其可变区的DNA作为模板并使用引物对的PCR(聚合酶链反应)方法进行扩增,以及使用编码引物对的肽接头的DNA的组合进行扩增,从而使所述肽接头的两端分别与重链或其可变区和轻链或其可变区连接)。
在本公开文本中,所述抗体或其抗原结合片段可以是人源化的。优选地,根据本发明的抗Ang2抗体可以是选自人抗体文库的完全人抗体,但不限于此。
根据本发明的抗体或其抗原结合片段的特征在于含有重链可变区,所述重链可变区包含具有SEQ ID NO:3的氨基酸序列的重链CDR1、具有SEQ ID NO:4的氨基酸序列的重链CDR2、具有SEQ ID NO:5的氨基酸序列的重链CDR3;以及轻链可变区,所述轻链可变区包含具有SEQ ID NO:6的氨基酸序列的轻链CDR1、具有SEQ ID NO:7的氨基酸序列的轻链CDR2、具有SEQ ID NO:8的氨基酸序列的轻链CDR3。
根据本发明的抗体或其抗原结合片段的特征在于含有重链可变区,所述重链可变区包含具有SEQ ID NO:13的氨基酸序列的重链CDR1、具有SEQ ID NO:14的氨基酸序列的重链CDR2、具有SEQ ID NO:15的氨基酸序列的重链CDR3;以及轻链可变区,所述轻链可变区包含具有SEQ ID NO:16的氨基酸序列的轻链CDR1、具有SEQ ID NO:17的氨基酸序列的轻链CDR2、具有SEQ ID NO:18的氨基酸序列的轻链CDR3。
在本发明中,所述抗体或其抗原结合片段的特征在于含有重链可变区,所述重链可变区包含SEQ ID NO:9、19、43、47、51、55、59、63、67、71、75、79、83、87、91、95、99、103、107或111的氨基酸序列;以及轻链可变区,所述轻链可变区包含SEQ ID NO:11、21、44、48、52、56、60、64、68、72、76、80、84、88、92、96、100、104、108或112的氨基酸序列,但不限于此。
所述抗体的氨基酸序列可以通过保守取代被取代。所述“保守取代”是指多肽的修饰,其包括至少一个氨基酸被具有与相应多肽类似的生物化学特性的氨基酸取代,而不引起生物学或生物化学功能的损失。“保守氨基酸取代”是指这样的取代,其中氨基酸残基被具有类似侧链的氨基酸残基替代。本领域定义了具有类似侧链的氨基酸残基种类。这些种类包括具有碱性侧链的氨基酸(例如赖氨酸、精氨酸、组氨酸)、具有酸性侧链的氨基酸(例如天冬氨酸、谷氨酸)、具有不带电荷的极性侧链的氨基酸(例如甘氨酸、天冬酰胺、谷氨酰胺、丝氨酸、苏氨酸、酪氨酸、半胱氨酸)、具有非极性侧链的氨基酸(例如丙氨酸、缬氨酸、亮氨酸、异亮氨酸、脯氨酸、苯丙氨酸、甲硫氨酸、色氨酸)、具有β-分支侧链的氨基酸(例如苏氨酸、缬氨酸、异亮氨酸)、以及具有芳香族侧链的氨基酸(例如酪氨酸、苯丙氨酸、色氨酸、组氨酸)。预期本发明的抗体在具有保守氨基酸取代的同时仍能够保留活性。
在本发明中,所述抗体或其抗原结合片段的特征在于含有由以保藏号KCLRF-BP-00417或KCLRF-BP-00418保藏的细胞系产生的抗体的互补决定区(CDR)。
本发明的抗Ang2抗体序列可以不同于本申请中提供的序列。例如,氨基酸序列与以上列出的那些的不同之处可在于:(a)可变区可以与轻链的恒定结构域分开,(b)氨基酸可以不同于以上列出的那些,而不会因此显著影响残基的化学特性(所谓的保守取代),(c)氨基酸可以与以上列出的那些相差给定百分比,例如,80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%或99%同源性。可替代地,编码抗体的核酸可以(a)与轻链的恒定结构域分开,(b)不同于以上列出的那些,而不会因此改变所编码的残基,或(c)可以与以上列出的那些相差给定百分比,例如,70%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%或99%同源性。
在进行氨基酸序列的保守改变时,可以考虑氨基酸的亲水指数。在本领域中通常理解亲水氨基酸指数在对蛋白质赋予相互作用性生物功能方面的重要性。公认的是,氨基酸的相对亲水特征有助于所得蛋白质的二级结构,继而定义了蛋白质与其他分子(例如酶、底物、受体、DNA、抗体、抗原等)的相互作用。
在本领域中还应理解,可以基于亲水性有效地进行相似氨基酸的取代。例如,蛋白质的如受其邻近氨基酸的亲水性管控的最大局部平均亲水性与所述蛋白质的生物特性相关。应理解,氨基酸可以被具有相似亲水性的另一种氨基酸取代并产生生物学或免疫学修饰的蛋白质。在此类变化中,亲水性值在+/-2之内的氨基酸的取代是优选的,在+/-1之内的那些氨基酸是特别优选的,并且在+/-0.5之内的那些氨基酸是甚至更特别优选的。
如上所述,氨基酸取代通常基于氨基酸侧链取代基的相对相似性,例如,它们的疏水性、亲水性、电荷、大小等。考虑了各种前述特征的示例性取代对于本领域技术人员是熟知的,并且包括:精氨酸和赖氨酸;谷氨酸和天冬氨酸;丝氨酸和苏氨酸;谷氨酰胺和天冬酰胺;以及缬氨酸、亮氨酸和异亮氨酸。
在另一方面,本发明涉及含有所述抗体或其抗原结合片段作为活性成分的药物组合物。
所述药物组合物的特征在于含有药学有效量的根据本发明的抗体或其抗原结合片段和药学上可接受的载体。
所述药物组合物可以进一步包含在化学疗法中使用的小分子抑制剂或血管内皮生长因子(VEGF)拮抗剂。所述VEGF拮抗剂可以是抗VEGF抗体、VEGF抑制性融合蛋白或小分子激酶抑制剂。
在另一方面,本发明涉及含有所述抗体或其抗原结合片段作为活性成分的用于预防或治疗眼病的药物组合物。
在另一方面,本发明涉及用于在眼病患者中抑制脉络膜新血管形成、抑制眼血管渗漏或同时触发脉络膜毛细血管再生的方法,所述方法包括向所述患者给予上述药物组合物。
用于预防或治疗眼病的药物组合物可以进一步包含在化学疗法中使用的小分子抑制剂或血管内皮生长因子(VEGF)拮抗剂。所述VEGF拮抗剂可以是抗VEGF抗体、VEGF抑制性融合蛋白或小分子激酶抑制剂。
所述方法可以进一步包括与本发明抗体或其片段的给予同时或逐步给予在化学疗法中使用的小分子抑制剂或血管内皮生长因子(VEGF)拮抗剂。所述VEGF拮抗剂可以是抗VEGF抗体、VEGF抑制性融合蛋白或小分子激酶抑制剂。
抗Ang2抗体或其抗原结合片段具有通过抑制Ang2的功能来抑制异常血管生成的功能,并且因此具有预防或治疗伴随血管异常的眼病的作用。
如本文所用,术语“预防”是指通过给予根据本发明的组合物来抑制或减慢眼病的进展的任何作用,并且术语“治疗”是指抑制、减轻或消除眼病的发展。
在本发明中,所述眼病是湿性年龄相关性黄斑变性(wAMD)、糖尿病性黄斑水肿(DME)或糖尿病性视网膜病(DR),但不限于此。
如本文所用,术语“黄斑变性”是指新血管形成异常生长,从而引起黄斑损伤并影响视力的病症。黄斑变性主要发生在超过50岁,并且分为非渗出性(干性)或渗出性(湿性)。特别是在湿性AMD的情况下,可能导致失明。AMD的病因尚未阐明,但已知危险因素是年龄;以及环境因素,包括吸烟、高血压、肥胖、遗传易感性、过度UV暴露、低血清抗氧化剂浓度等。
如本文所用,术语“黄斑水肿”是指视网膜黄斑的肿胀,并且所述肿胀是由于视网膜血管的流体渗漏而发生的。血液从薄弱的血管壁漏出,进入视网膜黄斑的局部区域,所述区域是颜色敏感的神经末梢,并且其中视网膜圆锥丰富。然后,图像淡化到中心的右侧或中心区域的中心。视力在几个月内逐渐降低。如本文所用,术语“糖尿病性视网膜病”是指眼睛的并发症,其中由于糖尿病引起的外周循环障碍引起的视网膜微循环障碍而导致视力下降。最初,它可能导致视力的轻度问题,但最终可能导致失明。糖尿病性视网膜病可以发生在患有1型糖尿病或2型糖尿病的任何人中。
本发明提供了包含治疗有效量的抗Ang2抗体和药学上可接受的载体的药物组合物。“药学上可接受的载体”是能够添加至活性成分中以帮助制剂的配制或稳定化的材料,并且它不对患者造成明显的不良毒理作用。
载体是指不抑制所给予化合物的生物学活性和特性并且不刺激患者的载体或稀释剂。待配制为液体溶液的组合物中的药学上可接受的载体被灭菌并且适于生命体。可以将盐水、无菌水、林格氏溶液、缓冲盐水、白蛋白注射液、右旋糖溶液、麦芽糖糊精溶液、甘油、乙醇用作载体,或者可以混合其至少一种组分以待使用,并且可以根据需要添其他常规添加剂,如抗氧化剂、缓冲液、抑菌剂等。此外,通过向其中进一步添加稀释剂、分散剂、表面活性剂、粘合剂和润滑剂,可以将组合物制备成注射剂(如水性溶液、悬浮液、乳剂等)、丸剂,胶囊、颗粒剂或片剂。其他载体是例如[Remington's Pharmaceutical Sciences(E.W.Martin)]中描述的。所述组合物可以含有治疗有效量的至少一种抗Ang2抗体。
药学上可接受的载体包括无菌水性溶液或分散体和用于制备临时无菌可注射溶液或分散体的无菌粉末。此类介质和药剂用于药物活性材料的用途是本领域已知的。优选地将组合物配制用于肠胃外注射。所述组合物可以被配制成溶液、微乳液、脂质体或适于高药物浓度的其他有序结构。载体可以是例如溶剂或分散介质,其含有水、乙醇、多元醇(例如,甘油、丙二醇和液体聚乙二醇等)及其适合的混合物。在一些情况下,所述组合物可以包含等渗剂(例如糖)、多元醇(如甘露糖醇、山梨糖醇)或氯化钠。可以通过将所需量的活性化合物与一种上述组分或其组合掺入适当的溶剂中,随后根据需要进行无菌微滤来制备无菌注射溶液。通常,通过将活性化合物掺入无菌媒介物中来制备分散体,所述无菌媒介物含有基本分散介质和来自上述组分的其他所需组分。通过真空干燥和冷冻干燥(冻干)活性成分粉末和任何另外所希望的组分粉末,从先前的无菌过滤溶液中获得用于制备无菌注射溶液的无菌粉末。
所述药物组合物可以口服或肠胃外给予,其剂量和频率可以取决于患者的严重程度而改变。可以根据需要通过推注或通过连续输注将所述组合物给予至患者。例如,作为Fab片段呈现的本发明抗体的推注给予可以具有0.0025至100mg/kg体重、0.025至0.25mg/kg、0.010至0.10mg/kg或0.10至0.50mg/kg的量。对于连续输注,可以将作为Fab片段呈现的本发明抗体以0.001至100mg/kg分钟、0.0125至1.25mg/kg/分钟、0.010至0.75mg/kg/分钟、0.010至1.0mg/kg/分钟或0.10至0.50mg/kg/分钟给予1至24小时、1至12小时、2至12小时、6至12小时、2至8小时或1至2小时。当给予作为全长抗体(具有完整的恒定区)呈现的本发明抗体时,给予量可以是约1至10mg/kg体重、2至8mg/kg或5至6mg/kg。通常通过持续30分钟至35分钟的注射来给予全长抗体。给予频率取决于病症的严重程度。所述频率可以是每周3次至一周一次或两周一次。
此外,可以通过皮下注射向患者给予所述组合物。例如,可以通过皮下注射每周、每两周或每月向患者给予具有10至100mg的给予量的抗Ang2抗体。
如本文所用,“治疗有效量”意指足以以适用于医学治疗的合理受益/风险比来治疗疾病的量,以及抗Ang2抗体的组合的量。精确量可以取决于包括治疗组合物的组分和物理特征、预期患者群体、个体患者考虑因素等的许多因素而改变,但不限于此,并且可以由本领域技术人员容易地确定。当完全考虑这些因素时,重要的是给予足以获得最大效果而没有副作用的最小量,并且此剂量可以由本领域的专家容易地确定。
本发明的药物组合物的剂量不受特别限制,但根据各种因素变化,所述因素包括患者的健康状况和体重、疾病的严重程度以及药物类型、给予途径和给予时间。可以将所述组合物以通常在哺乳动物(包括大鼠、小鼠、牛、人等)中允许的途径(例如口服、直肠、静脉内、皮下、子宫内或脑血管内)以每天单剂量量或多剂量来给予。
在另一方面,本发明涉及含有所述抗体或其抗原结合片段作为活性成分的用于预防或治疗癌症的药物组合物。
在另一方面,本发明涉及用于抑制患者中肿瘤生长并治疗癌症的方法,其包括向所述患者给予包含上述抗体或抗原结合片段的药物组合物。所述方法可以进一步包括与本发明抗体或其片段的给予同时或逐步给予在化学疗法中使用的小分子抑制剂或血管内皮生长因子(VEGF)拮抗剂。所述VEGF拮抗剂可以是抗VEGF抗体、VEGF抑制性融合蛋白或小分子激酶抑制剂。
如本文所用,术语“癌症”或“肿瘤”通常是指或描述以不受控制的细胞生长/增殖为特征的哺乳动物的生理状况。
可以用本发明的组合物治疗的癌症不受特别限制,并且包括实体癌和血液癌两者。此类癌症的例子包括鳞状细胞癌、小细胞肺癌、非小细胞肺癌、肺腺癌、肺鳞状细胞癌、腹膜癌、皮肤癌、皮肤或眼内黑色素瘤、直肠癌、肛门癌、食道癌、小肠癌、内分泌癌、甲状旁腺癌、肾上腺癌、软组织肉瘤、尿道癌、慢性或急性白血病、淋巴瘤、肝细胞癌、胃肠癌、胰腺癌、神经胶质瘤、宫颈癌、卵巢癌、肝癌、膀胱癌、肝肿瘤、乳腺癌、结肠癌、子宫内膜癌或子宫癌、唾液腺癌、肾癌、前列腺癌、外阴癌、甲状腺癌、头颈癌、脑癌、骨肉瘤等,但不限于此。
用于预防或治疗癌症的组合物包含抗Ang2抗体,并且其构成与用于预防或治疗眼部疾病的组合物中包含的组成相同,因此对每种构成的描述均等同地适用于预防或治疗癌症的组合物。
本申请还考虑与化学或放射治疗干预或其他治疗结合使用本文所述的抗Ang2抗体。具体地,将抗Ang2抗体与靶向Ang2功能的不同方面的其他疗法组合也可能证明是有效的。
在另一个实施方案中,本发明的抗体可以与至少一种药剂连接以形成抗体缀合物,以增加抗体分子作为诊断剂或治疗剂的功效。
在本发明的另一方面,本发明涉及编码所述抗体或其抗原结合片段的核酸。
本文所用的核酸可以存在于细胞、细胞裂解物中,或者还可以以部分纯化的形式或基本上纯的形式存在。当通过包括碱/SDS处理、CsCl显带、柱色谱、琼脂糖凝胶电泳和本领域熟知的其他技术的标准技术将核酸从其他细胞组分或其他污染物(例如,其他细胞核酸或蛋白质)纯化时,所述核酸是“分离的”或“基本上纯的”。本发明的核酸可以是例如DNA或RNA,并且可以包含内含子序列,或者可以不包含所述内含子序列。
在本发明的仍另一方面,本发明涉及包含所述核酸的重组表达载体。
为了表达所述抗体或其片段,可以通过标准分子生物学技术(例如,使用表达靶抗体的杂交瘤进行的PCR扩增或cDNA克隆)获得编码具有部分长度或全长的轻链和重链的DNA,并且所述DNA可以与待插入表达载体中的转录和翻译控制序列“可操作地结合”。
本文所用的术语“可操作地结合”可以指示抗体基因被连接到载体中,使得载体中的转录和翻译控制序列具有控制所述抗体基因的转录和翻译的预期功能。选择表达载体和表达控制序列,以具有与待使用的用于表达的宿主细胞的相容性。将抗体的轻链基因和抗体的重链基因插入单独的载体中,或将两种基因插入同一个表达载体中。通过标准方法(例如,抗体基因片段与载体上的互补限制酶位点进行连接,或者当完全不存在限制酶位点时进行平末端连接)将所述抗体插入表达载体中。在一些情况下,重组表达载体可以编码促进宿主细胞分泌抗体链的信号肽。可以将抗体链基因克隆到载体中,使得信号肽根据框架与抗体链基因的氨基末端结合。信号肽可以是免疫球蛋白信号肽或异源信号肽(即,源自除了免疫球蛋白之外的蛋白质的信号肽)。此外,重组表达载体具有调节序列,所述调节序列控制宿主细胞中抗体链基因的表达。“调节序列”可以包括控制抗体链基因的转录或翻译的启动子、增强子和其他表达控制元件(例如,聚腺苷酸化信号)。本领域技术人员能够认识到,可以根据因素(如待转化的宿主细胞的选择、蛋白质的表达水平等)通过改变调节序列来改变表达载体的设计。
在仍另一方面,本发明涉及用重组表达载体转化的细胞。
用于产生本公开文本的抗体的细胞可以是原核生物、酵母或高等真核细胞,但不限于此。
具体地,可以使用芽孢杆菌属(Bacillus,如大肠杆菌(Escherichia coli)、枯草杆菌(Bacillus subtilis)和土利真杆菌(Bacillus tuligensis))、链霉菌属(Streptomyces)、假单胞菌属(Pseudomonas,例如,恶臭假单胞菌(Pseudomonas putida))、以及原核宿主细胞如奇异变形杆菌(Proteus mirabilis)和葡萄球菌属(Staphylococcus,例如,肉葡萄球菌(Staphylococcus carnosus))的菌株。
对动物细胞的兴趣最大并且有用宿主细胞系的例子包括但不限于COS-7、BHK、CHO、CHOK1、DXB-11、DG-44、CHO/-DHFR、CV1、COS-7、HEK293、BHK、TM4、VERO、HELA、MDCK、BRL3A、W138、Hep G2、SK-Hep、MMT、TRI、MRC 5、FS4、3T3、RIN、A549、PC12、K562、PER.C6、SP2/0、NS-0、U20S或HT1080。
将核酸或载体转染到宿主细胞中。对于“转染”,可以使用各种普遍使用的技术(如电泳、磷酸钙沉淀、DEAE-葡聚糖转染、脂质转染等)将外源核酸(DNA或RNA)引入原核宿主细胞或真核宿主细胞中。考虑到适用于哺乳动物细胞中,根据本发明的抗体可以在真核细胞中表达,优选地在哺乳动物宿主细胞中表达。适于表达所述抗体的哺乳动物宿主细胞可以包括中国仓鼠卵巢(CHO)细胞(例如,包括与DHFR选择标记物一起使用的dhfr-CHO细胞)、NSO骨髓瘤细胞、COS细胞或SP2细胞等为例。
在另一方面,本发明涉及用于产生抗Ang2抗体或其抗原结合片段的方法,其包括培养宿主细胞并表达所述抗体或其抗原结合片段。
当将编码抗体基因的重组表达载体引入哺乳动物宿主细胞中时,所述抗体可以通过以下方法来产生:将宿主细胞培养足够的时间段以使抗体在宿主细胞中表达,或更优选地,培养足够的时间段以使抗体分泌到培养宿主细胞的培养基中。
在一些情况下,可以将表达的抗体与宿主细胞分离并纯化以获得均匀性。所述抗体的分离或纯化可以通过普遍用于蛋白质的分离方法、纯化方法(例如色谱法)来进行。所述色谱法可以包括例如,亲和色谱法、离子交换色谱法或包括蛋白A柱和蛋白G柱的疏水色谱法。除了色谱法之外,所述抗体还可以通过与过滤、超滤、盐析、透析等进一步组合来分离和纯化。
实施例
在下文中,将参考以下实施例更加详细地描述本发明。然而,以下实施例仅用于举例说明本发明,并且对于本领域技术人员而言清楚的是,本发明的范围不应解释为限制这些实施例。
实施例1:小鼠单克隆抗Ang2抗体的制备
1-1:用人Ang2进行小鼠免疫
为了用作抗原,将人Ang2(hAng2,SEQ ID NO:2)的受体结合结构域(RBD)克隆到含有CMV启动子的载体中,并通过转染到HEK293F细胞系中瞬时表达。在孵育5天后,将表达的重组人Ang2-RBD通过亲和柱进行纯化。将五周大的BALB/c小鼠每周用与佐剂混合的纯化的人Ang2-RBD(100μg/注射液)免疫两次,持续6周。通过hAng2 ELISA检查免疫小鼠的血清中的抗Ang2抗体滴度。当抗体滴度(1:5,000稀释度)适当地增加(OD>1.0)时,从免疫小鼠提取脾脏,并从中分离出B淋巴细胞并且将其与培养的骨髓瘤细胞(SP2/0)融合。在含有次黄嘌呤、氨基蝶呤和胸苷的HAT培养基中培养融合细胞,并从中选择仅由骨髓瘤细胞和B淋巴细胞的融合物构成的杂交瘤细胞并对其进行培养。将存活的杂交瘤细胞接种在96孔板中,并通过hAng2 ELISA测试培养上清液。通过有限稀释选择显示阳性信号的杂交瘤池用于克隆选择。最终,建立了约50个单克隆杂交瘤细胞系。其中有若干种Ang2结合抗体显示出Tie2激活活性。基于Tie2激活水平和对人Ang2的高亲和力选择候选抗体,然后进行处理以进行人源化。
[表1]
人血管生成素-2全长(hAng2)和受体结合结构域(RBD)序列
1-2:小鼠单克隆抗Ang2抗体的产生和纯化
为了产生基于ELISA阳性反应选择的抗Ang2抗体,在T75(75cm2面积)烧瓶中的含10%FBS的DMEM(杜尔贝科改良伊格尔培养基)中培养杂交瘤细胞。当细胞汇合度达到约90%时,将细胞用PBS洗涤,与50ml无血清培养基(SFM,Gibco)一起孵育并在37℃下培养3天。然后,收集从每种单克隆杂交瘤分泌抗体在其中的培养基,将其离心以去除细胞,并且将培养上清液收集并过滤。然后使用配备有Protein G亲和柱(GE Healthcare)的AKTA纯化设备(GE Healthcare)纯化抗体。使用离心过滤器单元(Amicon)通过用PBS代替上清液来浓缩纯化的抗体。
1-3:激活Tie2受体的抗Ang2抗体的鉴定和筛选
为了研究小鼠抗Ang2抗体是否诱导内皮细胞中Tie2受体的下游信号传导,用hAng2蛋白和抗Ang2抗体的组合处理HUVEC(Lonza),然后通过免疫印迹法分析Tie2受体的主要下游信号传导蛋白Akt磷酸化水平。作为阴性对照组,将单独的全长hAng2(R&Dsystems)处理到细胞中。
具体地,将HUVEC(1×105个细胞/ml)在60mm培养皿中于37℃下在EGM-2培养基(Lonza)中培养。将细胞(90%汇合度)与无血清EBM-2培养基孵育4小时,以进行血清饥饿。将血清饥饿的HUVEC用抗Ang2抗体和hAng2蛋白(1μg/ml,R&D system)的混合物处理,并进一步孵育30min。将细胞用冷PBS洗涤,用裂解缓冲液处理,并在4℃下裂解20min。然后,通过以13000rpm离心15min来制备细胞裂解物。将5x SDS样品缓冲液添加至细胞裂解物中,并将细胞裂解物在95℃下煮沸5分钟。然后,对细胞裂解物进行SDS PAGE,并将蛋白质转移至硝酸纤维素膜(GE)。
为了研究Akt磷酸化,将印迹用含5%脱脂乳的TBS-T在室温(RT)下封闭1hr,并与抗磷酸Akt抗体(S473)在4℃下孵育约8hr。通过增强的化学发光(ECL)可视化磷酸Akt的量。然后,将膜在剥离缓冲液(Thermo)中孵育15min,然后用抗Akt抗体再次探测以确定总Akt的量。
在分别用hAng2和抗Ang2抗体(如2C8、4B9、2F10和4E2)的组合处理的若干个组中,强烈诱导了S473处的Akt磷酸化(图1)。
1-4:通过Octet分析进行针对hAng2的抗Ang2抗体的亲和力测量
使用Octet系统(ForteBio)测量针对hAng2的小鼠单克隆抗体的亲和力。具体地,使用黑色96孔板(96孔F型黑色板,Greiner)以总计200μl/孔测量缓冲液和样品。将用于亲和力测量的生物传感器水合10min,然后使用AR2G尖端(ForteBio Octet)进行测量。在水合后,将hAng2以10μg/ml的浓度稀释在10mM乙酸钠缓冲液(pH 6.0)中,在AR2G生物传感器上固定,并用1M乙醇胺封闭。用1×动力学缓冲液将小鼠单克隆抗Ang2抗体稀释至50、25、12.5、6.25、3.125和0nM,并进行缔合300秒和解离900秒。对于亲和力测量值(KD),通过结合曲线(全局)分析缔合速率(K-on)和解离速率(K-off),并使用Octet数据分析v9.0.0.10程序将其拟合至1:1结合模型。KD值在下表2中示出。小鼠抗Ang2抗体对hAng2的亲和力在表2中示出。
[表2]
小鼠抗Ang2抗体对hAng2的亲和力
抗体 | Kon(1/Ms) | Koff(1/s) | K<sub>D</sub>(M) |
2C8 | 7.78E+04 | 3.54E-06 | 4.55E-11 |
2F10 | 1.24E+05 | 1.71E-05 | 1.38E-10 |
4B9 | 1.37E+05 | 5.04E-07 | 3.68E-12 |
4E2 | 2.83E+04 | 1.34E-04 | 4.74E-09 |
实施例2:小鼠抗Ang2抗体的DNA基因序列分析
分析了实施例1-3中选择的抗体(源自杂交瘤细胞)的DNA核苷酸序列。具体地,在含10%FBS的DMEM中培养杂交瘤细胞(2×106个细胞/ml),然后使用RNeasy迷你试剂盒(Qiagen)获得总RNA。接下来,测量RNA浓度,并通过逆转录(RT)反应合成cDNA。为了扩增每个杂交瘤细胞中产生的单克隆抗体的重链和轻链可变区基因序列,使用上述cDNA作为模板在以下条件下使用小鼠Ig-引物组(Novagen)进行PCR:94℃下5分钟;[94℃下1min,50℃下1min,72℃下2min]×35个循环;在72℃下6min;冷却至4℃。将从每个反应获得的PCR产物克隆到TA载体中,并进行DNA测序,从而获得编码每种抗体的CDR、重链可变区和轻链可变区的核苷酸序列(表3至表10)。
[表3]
小鼠抗Ang2抗体4B9的CDR序列
[表4]
小鼠抗Ang2抗体4B9的可变区序列
[表5]
小鼠抗Ang2抗体2C8的CDR序列
[表6]
小鼠抗Ang2抗体2C8的可变区序列
[表7]
小鼠抗Ang2抗体2F10的CDR序列
[表8]
小鼠抗Ang2抗体2F10的可变区序列
[表9]
小鼠抗Ang2抗体4E2的CDR序列
[表10]
小鼠抗Ang2抗体4E2的可变区序列
实施例3:针对hAng2的小鼠抗Ang2抗体的表位作图
通过HDX-MS(氢/氘交换质谱法)技术分析了由小鼠单克隆抗体2C8和4B9识别的hAng2的抗原决定簇(表位)。HDX-MS分析方法在以下文章中进行了描述;Houde D,Engen JR(2013)Methods Mol.Biol.988:269-89和Houde等人(2011)J.Pharm.Sci.100(6),2071。
将重组hAng2-RBD蛋白用于分析抗体2C8和4B9的表位。在氘标记反应之前,将hAng2-RBD/抗体混合物孵育多于3hr,以在15×稀释的氘标记缓冲液(KD=25nM)下保持最大结合(100%)。将制备的hAng2-RBD/抗体复合物用在不同时间标记的氘标记缓冲液稀释15倍,然后用相同体积的猝灭缓冲液猝灭。标记反应时间为0min(非氘)、0.33min、10min、60min和240min。然而,在非氘条件下,用平衡缓冲液替换氘标记缓冲液,并立即用猝灭缓冲液终止反应。对于质谱分析,将氘标记的hAng2-RBD/抗体样品加载至胃蛋白酶柱上,并进行肽消化。质谱分析显示,在hAng2-RBD的N末端处的13个肽和对应于25-40个氨基酸的消化性肽均没有被检测到,并且从总共45个消化性肽获得了83.7%的覆盖率数据。
比较地分析了单独的hAng2-RBD与hAng2-RBD-抗体复合物条件之间的氘摄取差异,并且显示出氘摄取明显降低的区域是抗体直接结合的肽或结构改变的区域。当单独的hAng2-RBD与hAng2-RBD-抗体复合物之间的氘摄取差异为0.5-1Da或更大时,其被认为是显著的。
氘摄取差异的分析指示,抗体2C8结合的表位是hAng2-RBD的SEQ ID NO:2的残基61至78-QRTWKEYKVGFGNPSGEY(SEQ ID NO:115)(表11),并且抗体4B9结合的表位是SEQ IDNO:2的残基14至24-KSGHTTNGIYT(SEQ ID NO:116)和SEQ ID NO:2的残基41至47-EAGGGGW(SEQ ID NO:117)(表12)。在抗体4B9的情况下,不能排除的是,未确定的区域(SEQ ID NO:2的残基25至40)可以被包括在所述表位的范围内。在使用PyMol软件生成的hAng2-RBD的3D结构上,每种抗体的表位分析结果以不同的颜色显示(图2)。
[表11]
通过HDX-MS对2C8与hAng2结合进行的表位作图分析
[表12]
通过HDX-MS对4B9与hAng2结合进行的表位作图分析
实施例4:小鼠抗Ang2抗体的人源化和全长IgG转化
为了消除小鼠抗Ang2抗体2C8和4B9在给予至人时的免疫原性,将抗体如下进行人源化。
4-1:重链人源化
与抗体2C8的重链序列显示出64%同源性的人抗体重链可变基因是IGHV1-46-01。基于这些分析,将2C8抗体的CDR区移植到人抗体重链可变基因IGHV1-46-01中。在此过程中,设计了5个人源化重链抗体基因(表13)。在人源化2C8的重链基因中引入了小鼠序列的反向突变,在表13的蛋白质序列中以粗体表示。
与抗体4B9的重链序列显示出80%同源性的人抗体重链可变基因是IGHV3-11-01。基于所述分析,将4B9抗体的CDR区移植到人抗体重链可变基因IGHV3-11-01中。作为结果,在此过程中设计了3个人源化重链抗体基因(表13)。在人源化4B9的重链基因中引入了小鼠序列的反向突变,在表13的蛋白质序列中以粗体表示。
4-2:轻链人源化
与抗体2C8的轻链序列显示出67%同源性的人抗体轻链可变基因是IGKV1-9-01。基于这些分析,将2C8抗体的CDR区移植到人抗体轻链可变基因IGKV1-9-01中。在此过程中设计了3个人源化轻链抗体基因(表13)。在人源化2C8的轻链基因中引入了小鼠序列的反向突变,在表13的蛋白质序列中以粗体表示。
与抗体4B9的轻链序列显示出70%同源性的人抗体轻链可变基因是IGKV1-39-01。基于这些分析,将4B9抗体的CDR区移植到人抗体轻链可变基因IGKV1-39-01中。在此过程中设计了1个人源化轻链抗体基因(表13)。
4-3:人源化基因合成和对人全长IgG抗体的克隆
将表15中的抗体的人源化可变区掺入人IgG1抗体的重链和轻链载体中。对应于抗体的人源化重链可变区(VH)的编码核苷酸由Bioneer,Inc.合成,从而组成“EcoRI-信号序列-VH-NheI-CH-XhoI”。对应于抗体的人源化轻链可变区(VL)的编码核苷酸由Bioneer,Inc.合成,从而组成“EcoRI-信号序列-VL-BsiWI-CL-XhoI”。使用EcoRI和XhoI建立表达全长人IgG抗体的载体,将编码重链的多核苷酸分别克隆到OptiCHOTM抗体表达试剂盒(Invitrogen)中包含的pOptiVECTM-TOPO TA克隆试剂盒的载体中,并将编码轻链的多核苷酸分别克隆到pcDNATM3.3-TOPO TA克隆试剂盒(Invitrogen)的载体中。对于构建各自命名为2C8H11G4和4B9H11G4的2C8H11和4B9H11的人IgG4类抗体,将2C8H11重链基因和4B9重链基因的恒定区(CH1-铰链-CH2-CH3)替换为编码IgG4类重链恒定区的多核苷酸。
[表13]
起源于小鼠4B9和2C8抗体的人源化抗Ang2抗体
4-4:人源化抗Ang2抗体的产生和纯化
为了产生人源化抗Ang2抗体,使用了能够以高效率产生重组蛋白的Expi293F(Gibco)细胞。在锥形瓶中培养Expi293F细胞(2×106个细胞/ml),并使用ExpiFectamine293转染试剂盒将编码重链和轻链的质粒共转染至Expi293F细胞中。将细胞在37℃,8%CO2下于振荡培养箱(定轨振荡器,125rpm)中培养5天。收集所得的培养基并将其离心以去除细胞。分离含有分泌的抗体的培养物上清液,并将其在4℃下储存,或立即使用配备有亲和柱(蛋白A琼脂糖柱,GE Healthcare)的AKTA纯化系统(GE Healthcare)纯化。通过使纯化的抗体通过0.2μm的蛋白质离心过滤器(Amicon)进行浓缩,同时用PBS替换溶液。
实施例5:针对hAng2的人源化抗Ang2抗体的亲和力测量
使用Octet系统(ForteBio)测量人源化抗Ang2抗体针对hAng2的亲和力。具体地,使用黑色96孔板(96孔F型黑色板,Greiner)以总计200μl/孔测量缓冲液和样品。将用于亲和力测量的生物传感器水合10min,然后使用AR2G尖端(ForteBio Octet)进行测量。在水合后,将人源化抗Ang2抗体以10μg/ml的浓度稀释在10mM乙酸钠缓冲液(pH 6.0)中,在AR2G生物传感器上固定,并用1M乙醇胺封闭。用1×动力学缓冲液将重组hAng2稀释至50、25、12.5、6.25、3.125和0nM,并进行缔合300秒和解离900秒。对于亲和力测量值(KD),通过结合曲线(全局)分析缔合速率(K-on)和解离速率(K-off),并使用Octet数据分析v9.0.0.10程序将其拟合至1:1结合模型。KD值在下表14-表15中示出。
人源化4B9抗体对hAng2的亲和力总结于表14中。人源化2C8抗体对hAng2的亲和力在表15中。此外,IgG4类2C8H11G4和4B9H11G4抗体也显示出与hAng2抗原的次纳摩尔高亲和力(表16)。
[表14]
人源化4B9抗体对hAng2的亲和力
抗体 | Kon(1/Ms) | Kdis(1/s) | K<sub>D</sub>(M) |
4B9H11 | 9.29E+04 | 1.58E-06 | 1.71E-11 |
4B9H21 | 7.37E+04 | 8.94E-06 | 1.21E-10 |
4B9H31 | 9.39E+04 | 1.56E-05 | 1.67E-10 |
[表15]
人源化2C8抗体对hAng2的亲和力
[表16]
IgG4类2C8H11G4和4B9H11G4抗体对hAng2的亲和力
抗体 | Kon(1/Ms) | Kdis(1/s) | K<sub>D</sub>(M) |
2C8H11G4 | 4.15E+05 | 4.35E-06 | 1.05E-11 |
4B9H11G4 | 4.32E+05 | 3.46E-05 | 8.00E-11 |
实施例6:所选择的人源化抗Ang2抗体的体外生物学特性分析
6-1:Akt磷酸化
为了研究人源化抗Ang2抗体是否在内皮细胞中诱导Tie2受体的下游信号传导,将HUVEC(Lonza)用人Ang2蛋白以及人源化抗Ang2抗体进行处理。然后,通过免疫印迹法测量Tie2受体的主要下游信号传导蛋白Akt磷酸化水平。为了比较Akt激活的程度,在实验中用单独的全长hAng2(R&D systems)或单独的抗体处理细胞。
具体地,将HUVEC细胞(1×105个细胞/ml)在60mm培养皿中于37℃下在EGM-2(Lonza)中培养。将90%汇合度的细胞与EBM-2(Lonza)一起孵育4hr。将血清饥饿的HUVEC用抗Ang2抗体和hAng2蛋白(1μg/ml,R&D system)的混合物处理,并进一步孵育30min。将细胞用冷PBS洗涤,用裂解缓冲液处理,并在4℃下裂解20min。然后,通过以13000rpm离心15min来制备细胞裂解物。将5x SDS样品缓冲液添加至细胞裂解物中,并将混合物在95℃下煮沸5min。然后,将混合物进行SDS-PAGE和随后的蛋白质印迹。
为了研究Akt的磷酸化,将膜用含5%脱脂乳的TBST在RT下封闭1hr,并与抗磷酸Akt抗体(S473)在4℃下孵育约8hr。通过增强的化学发光(ECL)可视化磷酸Akt的量。然后,将膜在剥离缓冲液(Thermo)中孵育15min,然后用抗Akt抗体再次探测以确定总Akt的量。
如图3所示,通过在存在hAng2的情况下用0.5μg/ml抗Ang2抗体处理,Akt磷酸化显著增加,并且其在4B9H11处理组和2C8H11处理组两者中均维持直到50μg/ml的抗体浓度。这些数据指示人源化抗Ang2抗体能够在内皮细胞中强烈诱导Tie2受体的主要下游信号传导分子Akt的激活。当测试人源化4B9H11-或2C8H11-IgG4抗体时,观察到类似的模式。
6-2:由人源化抗Ang2抗体诱导的Tie2磷酸化
Ang2与Tie2受体结合并充当弱激动剂或拮抗剂。本发明中开发的抗Ang2抗体与Ang2结合以诱导Ang2-抗体复合物,从而进一步引起Tie2受体的聚集并因此增强Tie2受体的激活。进行实验以使用HUVEC分析抗Ang2抗体对Tie2磷酸化的影响。
具体地,将HUVEC(Lonza)在100mm培养皿中于37℃和5%CO2浓度下在EGM-2(Lonza)中培养。在80%-90%汇合度时,将细胞更换至EBM-2(Lonza)培养基持续2hr至6hr以进行血清饥饿。将各种浓度(0.02μg/ml至50μg/ml)的人源化抗Ang2抗体与hAng2蛋白(1μg/ml,R&D systems)混合30min。然后,将混合物用培养的细胞处理并进一步孵育30min。将细胞用冷PBS洗涤两次,然后在1000μl裂解缓冲液(10mM Tris-Cl(pH 7.4)、150mM NaCl、5mM EDTA、10%甘油、1%Triton X-100、蛋白酶抑制剂、磷酸酶抑制剂)中裂解,然后在4℃下裂解60min。制备细胞提取物,并将其以12,000rpm离心10min。通过BCA测定定量上清液。
向0.5mg细胞裂解物中添加1μg的Tie2抗体(R&D systems,AF313),并将其在4℃下孵育过夜。然后,添加DynabeadsTM蛋白G(Life technologies)以反应2hr,并进行免疫沉淀。使用磁铁将珠固定在试管的一侧上,用裂解缓冲液洗涤三次,然后在70℃下与含还原剂的2x SDS样品缓冲液一起孵育10min。从样品去除珠,并在4%-15%SDS蛋白凝胶(Bio-Rad)上电泳,然后转移至0.45μm PVDF膜。
在室温下用与5%(v/v)BSA混合的TBS-T将膜封闭1hr,并在4℃下与抗磷酸酪氨酸抗体(4G10,Millipore)孵育8hr,接着孵育HRP缀合的抗小鼠抗体并随后进行蛋白质印迹。为了测量免疫沉淀的Tie2的量,使膜在剥离缓冲液(Thermo)中反应15min,然后再次封闭并用抗Tie2抗体(R&D systems,AF313)再次探测。如图4所示,当将抗Ang2抗体与Ang2一起添加至HUVEC细胞时,以剂量依赖性方式强烈诱导了Tie2的磷酸化,与图3中类似。当测试人源化4B9H11-或2C8H11-IgG4抗体时,观察到类似的模式。这些数据指示人源化抗Ang2抗体2C8H11和4B9H11直接诱导人内皮细胞中Tie2受体的激活。
6-3:HUVEC中的Tie2聚集和FOXO1易位
在HUVEC中通过免疫荧光检测了通过抗Ang2抗体导致的在细胞间连接区域处的Tie2聚集和FOXO1从细胞核到胞质溶胶的易位。具体地,将HUVEC接种在8孔载玻片室(Lab-TekII)上,并在EGM-2培养基中保持2至3天。在100%汇合度时,将细胞用EBM-2培养基进行血清饥饿4hr,然后用1μg/ml抗Ang2抗体以及1μg/ml hAng2处理30min。此后,将细胞在室温(RT)下用PBS中的4%甲醛固定10min,用PBS中的0.1%Triton X-100进行透性化处理,在RT下用PBS中的1%BSA封闭60min,并且在RT下与一抗孵育1hr。使用了hTie2、FOXO1和人Fc的一抗。然后将细胞与二抗(Invitrogen)在黑暗中于RT下孵育1hr,并用具有DAPI的Vectashield封片剂(Vector Labs)进行封固。用激光扫描共聚焦显微镜(LSM880,CarlZeiss)拍摄图像。
如图5所示,用2C8H11或4B9H11与hAng2一起处理诱导了Tie2易位/聚集至细胞间接触处(cell-cell contact),就像已知诱导Tie2聚集和激活的Comp-Ang1(CA1)或对照Ang2抗体一样(Han等人,2016,Science Translation Medicine)。与显示FOXO1在磷酸化后定位在细胞质中的先前报道(Zhang等人,JBC 2002,277,45276–45284)一致,当FOXO1在基础的、血清饥饿的条件下定位在细胞核中时,与血清饥饿的对照(红色)相比,在用2C8H11+hAng2或4B9H11+hAng2处理的情况下FOXO1在细胞核中明显消失。同时,Ang2处理可忽略不计地诱导FOXO1从细胞核易位至胞质溶胶。有趣的是,发现2C8H11、4B9H11(青色)人源化抗体与聚集的Tie2受体共定位在细胞间接触处,并且所述抗体在胞质溶胶中内吞Tie2受体(图5),这指示抗Ang2抗体通过结合至Ang2而与Tie2受体形成三重复合物。
在时间进程研究(从10min至240min)中,检查了2C8H11诱导的Tie2聚集和FOXO1易位。如图6所示,在存在hAng2的情况下,对照Ang2 Ab在10min内诱导了在细胞间接触处(绿色)的Tie2聚集,并触发了聚集的Tie2受体的内吞作用。在用对照Ang2 Ab+hAng2处理30min后,细胞间接触处的Tie2受体明显减少,并且Tie2受体在120min和240min内大部分消失。当用抗人Fc抗体(青色)染色对照Ang2抗体时,其显示出如同Tie2受体那样的类似图案。相比之下,在2C8H11和hAng2的情况下,即使在240min处理后,细胞间接触处的Tie2聚集仍得以维持。一致地,在细胞间接触处与Tie2共定位的2C8H11抗体也被维持直到240min(图6)。
6-4:通过人源化抗Ang2抗体抑制血管通透性
根据制造商的说明,使用体外血管通透性测定试剂盒(Millipore)在HUVEC中进行了血管渗漏测定。将HUVEC接种到穿透板的插入物中,并培养3天以达到100%汇合度。将HUVEC与Ang2(1μg/ml)、Ang2(1μg/ml)以及对照、2C8H11或4B9H11抗体(1μg/ml)一起预孵育30min,然后添加TNF-a(100ng/ml),并将细胞在37℃下孵育22hr。将FITC-葡聚糖添加至上部室中并孵育20min。由荧光读取器分别在485和535nm的激发和发射波长下测量FITC-葡聚糖穿过HUVEC单层的通过。如图7所示,用抗Ang2抗体与Ang2预处理显著地抑制了由血管渗漏促进因子TNF-a诱导的血管渗漏。
实施例7:针对mAng2的人源化抗Ang2抗体的亲和力测量
通过ELISA分析人源化抗体对小鼠Ang2(mAng2)的亲和力。具体地,在半96孔板(Corning 3690)中以每孔20ng将mAng2稀释在30μl包被缓冲液(0.1M碳酸钠缓冲液)中,并在4℃下孵育过夜。在用TBS-T溶液洗涤3次后,在室温下将孔板用3%脱脂乳封闭1hr,然后再次洗涤。将2C8H11和4B9H11从3mg/ml连续稀释至300ng/ml。在将30μl稀释的抗Ang2抗体加载至孔中后,将孔板在室温下孵育2hr。接下来,将30μl的抗人IgG(Fab)-HRP(Jackson)二抗1:3000稀释液添加至每个孔中,并在室温下孵育1hr。在所有反应完成后,将板用TBS-T再次洗涤,然后每个孔用30μl的TMB溶液处理。在显影5min后,用1N硫酸处理板以终止反应,并在450nm处测量吸光度。基于所测量的OD值,使用PerkinElmer的WorkOut 2.5程序分析EC50值。4B9H11和2C8H11对于mAng2结合的EC50分别为105μg/ml和97μg/ml(图8)。
实施例8:LLC皮下模型中肿瘤生长抑制作用的评价。
在LLC(Lewis Lung Carcinoma)细胞系肿瘤模型中测试了2C8H11抗Ang2抗体抑制肿瘤生长的能力。具体地,将LLC细胞系(ATCC)在补充有10%FBS(Gibco)的DMEM(Gibco)中培养。将LLC细胞(在100μl PBS中的1×106个)皮下注射到用氯胺酮和甲苯噻嗪的混合物麻醉的6至8周大C57BL/6小鼠(Jackson Laboratory)中。当肿瘤体积达到50至100mm3时,每2至3天向小鼠腹膜内给予10mg/kg的2C8H11抗体。在单一疗法和组合疗法组两者中,以3mg/kg的剂量腹膜内注射一次顺铂(Cpt)。在接下来的几天中追踪肿瘤体积的变化。使用以下公式测量肿瘤体积(V):
V=(宽度2×长度)/2
在4个组中进行实验:Fc(对照组)、Fc+Cpt组、2C8H11组和2C8H11+Cpt组。如图9所示,与Fc相比,2C8H11抗体将肿瘤生长抑制了29%,这类似于通过Fc+Cpt注射所获得的肿瘤生长抑制作用。同时,与Fc相比,用2C8H11和Cpt组合处理使肿瘤生长延迟47%。因此,这些结果证明,用2C8H11与Cpt组合处理最有效地抑制了肿瘤生长。
实施例9:2C8H11抗体的肿瘤血管正常化作用。
为了研究通过2C811抗体所导致的肿瘤血管的变化,我们获得了肿瘤样品的冷冻切片,并通过用血管特异性标记物CD31和周细胞特异性标记物PDGFRβ染色进行了免疫荧光分析。具体地,从来自实施例8中描述的实验的小鼠收获肿瘤样品,将其固定在4%多聚甲醛(PFA,Merck)中,在30%蔗糖(Junsei)中脱水,包埋在OCT化合物(Leica)中,并使用低温恒温器(Leica)进行切片。使用蛋白质封闭缓冲液(DAKO)将所得冷冻切片封闭1hr。然后在4℃下将切片用PBS中的仓鼠抗CD31抗体(1:200,Millipore)和大鼠抗PDGFRβ抗体(1:200,eBioscience)染色8hr。在用PBS洗涤3次后,在室温下将切片用PBS中的Alexa488缀合的抗仓鼠IgG抗体和Alexa594缀合的抗大鼠IgG抗体(1:1000,Jackson Immunoresearch)染色1hr。在用PBS另外洗涤3次后,使用盖玻片(Marienfeld)将切片封固在荧光封片剂(DAKO)中。使用LSM880共聚焦显微镜(Zeiss)将染色的切片成像。
结果在图10中示出;红色信号指示CD31+区域,并且绿色信号指示PDGFR-β+区域。与用Fc或Fc+Cpt处理的肿瘤相比,在2C8H11或2C8H11+Cpt处理的肿瘤中,肿瘤血管(BV)密度降低了56%,并且这些脉管的形态已正常化为与正常血管相似。此外,用2C8H11或2C8H11+Cpt处理的肿瘤具有增加的PDGFRβ+周细胞覆盖率(增加2.4倍),并且血管和血管周细胞彼此更密切地相关。这些结果显示2C8H11抗体可以降低肿瘤块内的血管密度并使其形态正常化。
实施例10:通过2C8H11抗体增加肿瘤血管的功能性。
为了分析用2C8H11处理后肿瘤血管的功能性,评价了血管灌注性(perfusability)和缺氧状态。在收获肿瘤块之前,向小鼠静脉注射100μl DyLight488-凝集素(Vector laboratory),并腹膜内注射60mg/kg溶解在PBS中的盐酸哌莫硝唑(Hypoxyprobe)30min,然后处死。用4%PFA灌注固定小鼠。我们从肿瘤块获得冷冻切片,将其用PBS中的仓鼠抗CD31抗体(1:200,Millipore)和4.3.11.3小鼠Pacific blue-Mab(1:50,Hypoxyprobe)染色。使用LSM880共聚焦显微镜(Zeiss)将切片成像,并使用ImageJ软件(http://rsb.info.nih.gov/ij)分析获得的图像,以定量凝集素+面积/CD31+面积和低氧探针+面积。
结果在图11中示出。与用Fc或Fc+Cpt处理的肿瘤相比,用2C8H11或2C8H11+Cpt处理的肿瘤显示出灌注增强的正常化肿瘤血管,如通过增加的凝集素+面积(绿色)/CD31+面积(红色)(灌注增加大约3倍)所判断。此外,当与用Fc或Fc+Cpt处理的肿瘤相比时,在用2C8H11或2C8H11+Cpt处理的肿瘤中的如由低氧探针+面积(绿色)所指示的缺氧减少了72%。这些结果指示2C8H11抗体不仅使肿瘤血管的形态正常化,而且通过增加血管灌注性(这随后导致缺氧减少)来增强其功能性。
实施例11:通过2C8H11抗体增加抗癌药物向肿瘤块中的递送。
所述药物Cpt通过抑制DNA合成来抑制肿瘤生长,并且已广泛用于人癌症患者。为了评价2C8抗体是否可以通过使肿瘤血管正常化来增加此药物向肿瘤中的递送,将肿瘤的冷冻切片用抗顺铂修饰的DNA抗体(1:100,Abcam)和仓鼠抗CD31抗体(1:200,Millipore)进行染色。如图12所示,与Fc+Cpt处理组相比,在2C8H11/Cpt处理组中Cpt修饰的DNA(绿色)水平显著增加了2.1倍。此结果显示Cpt向肿瘤块的递送由于通过2C8H11抗体使肿瘤血管正常化而增强,这随后有效地抑制了肿瘤生长。
实施例12:激光诱导的CNV模型中2C8H11抗体的CNV消退和血管渗漏抑制作用。
使用激光诱导的CNV模型测试2C8H11抗体抑制脉络膜新血管形成(CNV)的能力,所述脉络膜新血管形成是湿性年龄相关性黄斑变性(AMD)的标志。在用5mg/ml去氧肾上腺素和5mg/ml托吡卡胺滴眼剂(Santen Pharmaceutical)扩张瞳孔,并滴注0.5%盐酸丙美卡因滴眼剂(Alcon)用于局部麻醉后,用玻璃盖玻片作为接触镜片,将具有裂隙灯递送系统的激光光凝器(Lumenis Inc.)用于可视化视网膜。对于每只眼睛在4个位置(后极的3、6、9和12点钟位置)递送足够的激光能量(532nm波长、250mW功率、100ms持续时间、50μm光斑大小)。此研究仅包括在激光光凝时产生气泡(指示布鲁赫膜(Bruch’s membrane)破裂)的烧伤。在激光部位处含有出血的光斑被排除在分析之外。为了概括临床情况,在激光光凝后7天,向小鼠玻璃体内给予2C8H11(5μg)(图13A)。作为对照或用于比较,以相同的方式向小鼠给予Fc或VEGF-Trap(各5μg)。为了玻璃体内给予指示的试剂,使用配有玻璃毛细管移液管的Nanoliter 2000微型注射器(World Precision Instruments)将含有5μg每种试剂的约1μl(5mg/ml)注射到玻璃体腔内。在激光光凝后14天,使用MATLAB图像处理工具箱(MathWorks)计算视网膜色素上皮细胞(RPE)-脉络膜-巩膜铺片的CD31+CNV体积。将抗CD31抗体(1:200,Millipore)用于检测CNV的内皮细胞。与Fc相比,VEGF-Trap有效地诱导CNV消退了64.4%,并且2C8H11类似地诱导了CNV消退(65.3%)(图13B、图13C)。组合的荧光素血管造影术(FA)和吲哚菁绿血管造影术(ICGA)使我们能够测量激光损伤部位周围新生血管处的血管渗漏。将488nm和785nm处的连续波激光模块分别用作荧光素和ICG的激发源。通过由旋转多面镜(MC-5;Lincoln Laser)和基于检流计的扫描镜(6230H;Cambridge technology)组成的扫描仪系统获得激发激光器的光栅扫描图案,并将其传送到成像透镜的后孔。将高数值孔径(NA)物镜(PlanApoλ,NA 0.75;Nikon)用作成像透镜以提供广角眼底荧光图像。通过帧捕获器将由光电倍增管(R9110;Hamamatsu Photonics)检测到的荧光信号数字化,并实时重建为具有每帧512×512像素大小的图像。为了利用血管造影术系统可视化晚期(6min)FA和ICGA图像,分别通过腹膜内和静脉内给予10mg荧光素钠(Alcon)和0.15mg ICG(DaiichiPharmaceutical)。在全身麻醉和瞳孔扩张下进行成像程序,以提高图像质量。使用基于Java的成像软件(ImajeJ;National Institutes of Health)将CNV的渗漏面积计算为FA图像中的总测量超荧光面积除以ICGA图像中的总测量CNV面积。与Fc相比,VEGF-Trap(37.0%)和2C8H11(38.3%)两者均类似地抑制了血管渗漏(图13B、图13D)。值得注意的是,Fc处理组在激光光凝后6天与14天之间显示出血管渗漏无显著差异,而VEGF-Trap和2C8H11显著减少了血管渗漏(分别为45.6%和50.0%)(图13B、图13D)。因此,在激光诱导的CNV小鼠模型中,VEGF-Trap与2C8H11之间CNV和血管渗漏的抑制程度在定量上是无法区分的。
实施例13:2C8H11抗体的CNV消退和脉络膜毛细血管再生作用。
为了确定2C8H11在CNV建立后在CNV消退和脉络膜毛细血管再生方面的作用,在激光光凝后7天,通过Nanoliter 2000微型注射器(World Precision Instruments)向小鼠玻璃体内给予Fc、VEGF-Trap、对照抗体或2C8H11(各5μg)。在激光光凝后6、14、21和35天进行了活体光学相干断层扫描血管造影术(OCTA)(图14A)。以230kHz的A扫描速率,使用利用以1048nm为中心的定制环形腔波长扫描激光器的原型高速扫描源光学相干断层扫描术(OCT)系统将视网膜脉络膜层成像。在视网膜脉络膜层内的1.7mm x 1.7mm视野中收集OCT图像,以监测玻璃体内注射试剂后激光光凝部位处脉络膜脉管系统的再生。为了获得截面OCT血管造影片(其允许在没有视网膜和脉络膜实质的情况下选择性地可视化血管),我们比较了重复记录的B扫描图像,并检测了主要由血管内红细胞运动引起的那些图像的逐像素强度去相关性。然后,通过使用自动的层展平和分段算法,将截面OCT血管造影片展平为RPE,并通过在三个深度范围:内视网膜、外视网膜和脉络膜层内对每个展平的截面OCT血管造影片进行单独投影来生成正面OCT血管造影片。将外丛状层和布鲁赫膜定义为分隔内视网膜、外视网膜和脉络膜层的边界。将视网膜和脉络膜血管的密度自动计算为被流动血管占据的测量面积的比例,所述流动血管被定义为具有高于阈值水平的去相关值的像素。借助于MATLAB(MathWorks)的图像处理工具箱,从代表脉络膜层的正面OCT血管造影片检测出无血管像素。然后,通过将无血管像素的数量乘以一个像素的体积,计算激光损伤部位周围的无血管空间的总体积。为了分析无血管空间体积的变化情况,将每只眼睛的连续测量值转化为相对于基线值的百分比变化。用Fc处理的外视网膜中的CNV体积略微减少,但用VEGF-Trap和2C8H11处理的那些外视网膜显示CNV体积明显减少(图14B、图14C)。同时,在用Fc处理的脉络膜中观察到无血管空间的略微减少。有趣的是,在激光光凝后14、21和35天,经2C8H11处理的眼睛中的脉络膜分别显示无血管空间连续且明显减少了30.1%、36.4%和37.0%(图14B、图14D)。类似地,在激光光凝后14、21和35天,经对照Ab处理的眼睛中的脉络膜分别显示无血管空间减少了21.7%、30.2%和38.0%(图14B、图14D)。然而,在D14、D21和D35,经VEGF-Trap处理的眼睛中的脉络膜分别显示无血管空间增加了11.4%、16.0%和18.1%(图14B、图14D)。总体而言,这些发现指示在激光诱导的CNV模型中,2C8H11和对照Ab两者均促进脉络膜毛细血管层的再生,而VEGF-Trap导致脉络膜毛细血管消退。
实施例14:2C8H11抗体和CD31在CNV内皮细胞中的共定位。
为了研究皮下注射的2C8H11是否也可以对CNV发挥治疗作用,我们首先评价了2C8H11抗体和CD31在CNV内皮细胞中的共定位。在激光光凝后1天进行2C8H11抗体(25mg/kg)的皮下给予。作为对照,以相同的方式向小鼠给予Fc(25mg/kg)。在激光光凝后2、4和8天,通过抗人IgG抗体(1:1000,Jackson ImmunoResearch Laboratories)直接检测2C8H11抗体和抗CD31抗体(1:200,Millipore)在CNV内皮细胞中的共定位(图15A)。给予的2C8H11在CNV区域中的CD31+内皮细胞中高度可检测(图15B-图15D)。
实施例15:皮下注射的2C8H11抗体的CNV抑制作用。
为了确定皮下注射的2C8H11抗体在CNV抑制方面的作用,在激光光凝后1天进行2C8H11抗体(25mg/kg)的皮下给予。作为对照,以相同的方式向小鼠给予Fc(25mg/kg)。将抗CD31抗体(1:200,Millipore)用于检测CNV的内皮细胞,并在激光光凝后8天使用MATLAB图像处理工具箱(MathWorks)计算RPE-脉络膜-巩膜铺片的CD31+CNV体积(图16A)。与Fc相比,2C8H11有效地将CNV形成抑制了69.9%(图16B、图16C),这指示2C8H11的玻璃体内注射和皮下注射均具有对CNV的抑制作用。
将本发明的微生物命名为2C8,并于2018年1月30日保藏于韩国首尔钟路区仰光洞28号110-744,首尔国立大学医学院的癌症研究所(Cancer Research Institute)的韩国细胞系库(Korean Cell Line Bank,KCLB)(保藏号:KCLRF-BP-00417)。
将本发明的微生物命名为4B9,并于2018年1月30日保藏于韩国首尔钟路区仰光洞28号110-744,首尔国立大学医学院的癌症研究所(Cancer Research Institute)的韩国细胞系库(Korean Cell Line Bank,KCLB)(保藏号:KCLRF-BP-00418)。
工业实用性
本发明涉及抑制Ang2并同时激活Tie2受体从而导致促进下游信号转导的抗体。它提供了抑制Ang2诱导的血管生成并降低血管通透性的方法。此外,根据本发明的抗体可以用于诊断和治疗异常的与血管生成有关的疾病,如眼部疾病或癌症和/或由增加的血管通透性引起的疾病。
已基于本发明的特定特征对其进行了详细描述,并且对于本领域技术人员而言明显的是,这些特定技术仅是较好的实施方案,并且因此本发明的范围不限于所述实施方案。因此,本发明的实质范围将由所附权利要求及其等同物来限定。
序列表自由文本
附在电子文件中。
<110> 基础科学研究院
韩国科学技术院
<120> 抗血管生成素-2抗体及其用途
<130> PP-B2174
<150> US 62/633,038
<151> 2018-02-20
<150> KR 10-2019-0018769
<151> 2019-02-18
<160> 117
<170> KoPatentIn 3.0
<210> 1
<211> 496
<212> PRT
<213> 人工序列
<220>
<223> 人血管生成素-2全长
<400> 1
Met Trp Gln Ile Val Phe Phe Thr Leu Ser Cys Asp Leu Val Leu Ala
1 5 10 15
Ala Ala Tyr Asn Asn Phe Arg Lys Ser Met Asp Ser Ile Gly Lys Lys
20 25 30
Gln Tyr Gln Val Gln His Gly Ser Cys Ser Tyr Thr Phe Leu Leu Pro
35 40 45
Glu Met Asp Asn Cys Arg Ser Ser Ser Ser Pro Tyr Val Ser Asn Ala
50 55 60
Val Gln Arg Asp Ala Pro Leu Glu Tyr Asp Asp Ser Val Gln Arg Leu
65 70 75 80
Gln Val Leu Glu Asn Ile Met Glu Asn Asn Thr Gln Trp Leu Met Lys
85 90 95
Leu Glu Asn Tyr Ile Gln Asp Asn Met Lys Lys Glu Met Val Glu Ile
100 105 110
Gln Gln Asn Ala Val Gln Asn Gln Thr Ala Val Met Ile Glu Ile Gly
115 120 125
Thr Asn Leu Leu Asn Gln Thr Ala Glu Gln Thr Arg Lys Leu Thr Asp
130 135 140
Val Glu Ala Gln Val Leu Asn Gln Thr Thr Arg Leu Glu Leu Gln Leu
145 150 155 160
Leu Glu His Ser Leu Ser Thr Asn Lys Leu Glu Lys Gln Ile Leu Asp
165 170 175
Gln Thr Ser Glu Ile Asn Lys Leu Gln Asp Lys Asn Ser Phe Leu Glu
180 185 190
Lys Lys Val Leu Ala Met Glu Asp Lys His Ile Ile Gln Leu Gln Ser
195 200 205
Ile Lys Glu Glu Lys Asp Gln Leu Gln Val Leu Val Ser Lys Gln Asn
210 215 220
Ser Ile Ile Glu Glu Leu Glu Lys Lys Ile Val Thr Ala Thr Val Asn
225 230 235 240
Asn Ser Val Leu Gln Lys Gln Gln His Asp Leu Met Glu Thr Val Asn
245 250 255
Asn Leu Leu Thr Met Met Ser Thr Ser Asn Ser Ala Lys Asp Pro Thr
260 265 270
Val Ala Lys Glu Glu Gln Ile Ser Phe Arg Asp Cys Ala Glu Val Phe
275 280 285
Lys Ser Gly His Thr Thr Asn Gly Ile Tyr Thr Leu Thr Phe Pro Asn
290 295 300
Ser Thr Glu Glu Ile Lys Ala Tyr Cys Asp Met Glu Ala Gly Gly Gly
305 310 315 320
Gly Trp Thr Ile Ile Gln Arg Arg Glu Asp Gly Ser Val Asp Phe Gln
325 330 335
Arg Thr Trp Lys Glu Tyr Lys Val Gly Phe Gly Asn Pro Ser Gly Glu
340 345 350
Tyr Trp Leu Gly Asn Glu Phe Val Ser Gln Leu Thr Asn Gln Gln Arg
355 360 365
Tyr Val Leu Lys Ile His Leu Lys Asp Trp Glu Gly Asn Glu Ala Tyr
370 375 380
Ser Leu Tyr Glu His Phe Tyr Leu Ser Ser Glu Glu Leu Asn Tyr Arg
385 390 395 400
Ile His Leu Lys Gly Leu Thr Gly Thr Ala Gly Lys Ile Ser Ser Ile
405 410 415
Ser Gln Pro Gly Asn Asp Phe Ser Thr Lys Asp Gly Asp Asn Asp Lys
420 425 430
Cys Ile Cys Lys Cys Ser Gln Met Leu Thr Gly Gly Trp Trp Phe Asp
435 440 445
Ala Cys Gly Pro Ser Asn Leu Asn Gly Met Tyr Tyr Pro Gln Arg Gln
450 455 460
Asn Thr Asn Lys Phe Asn Gly Ile Lys Trp Tyr Tyr Trp Lys Gly Ser
465 470 475 480
Gly Tyr Ser Leu Lys Ala Thr Thr Met Met Ile Arg Pro Ala Asp Phe
485 490 495
<210> 2
<211> 221
<212> PRT
<213> 人工序列
<220>
<223> 人血管生成素-2受体结合结构域(RBD)
<400> 2
Glu Glu Gln Ile Ser Phe Arg Asp Cys Ala Glu Val Phe Lys Ser Gly
1 5 10 15
His Thr Thr Asn Gly Ile Tyr Thr Leu Thr Phe Pro Asn Ser Thr Glu
20 25 30
Glu Ile Lys Ala Tyr Cys Asp Met Glu Ala Gly Gly Gly Gly Trp Thr
35 40 45
Ile Ile Gln Arg Arg Glu Asp Gly Ser Val Asp Phe Gln Arg Thr Trp
50 55 60
Lys Glu Tyr Lys Val Gly Phe Gly Asn Pro Ser Gly Glu Tyr Trp Leu
65 70 75 80
Gly Asn Glu Phe Val Ser Gln Leu Thr Asn Gln Gln Arg Tyr Val Leu
85 90 95
Lys Ile His Leu Lys Asp Trp Glu Gly Asn Glu Ala Tyr Ser Leu Tyr
100 105 110
Glu His Phe Tyr Leu Ser Ser Glu Glu Leu Asn Tyr Arg Ile His Leu
115 120 125
Lys Gly Leu Thr Gly Thr Ala Gly Lys Ile Ser Ser Ile Ser Gln Pro
130 135 140
Gly Asn Asp Phe Ser Thr Lys Asp Gly Asp Asn Asp Lys Cys Ile Cys
145 150 155 160
Lys Cys Ser Gln Met Leu Thr Gly Gly Trp Trp Phe Asp Ala Cys Gly
165 170 175
Pro Ser Asn Leu Asn Gly Met Tyr Tyr Pro Gln Arg Gln Asn Thr Asn
180 185 190
Lys Phe Asn Gly Ile Lys Trp Tyr Tyr Trp Lys Gly Ser Gly Tyr Ser
195 200 205
Leu Lys Ala Thr Thr Met Met Ile Arg Pro Ala Asp Phe
210 215 220
<210> 3
<211> 5
<212> PRT
<213> 人工序列
<220>
<223> CDRH1
<400> 3
Asp Tyr Tyr Met Tyr
1 5
<210> 4
<211> 17
<212> PRT
<213> 人工序列
<220>
<223> CDRH2
<400> 4
Thr Ile Ser Val Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val Lys
1 5 10 15
Gly
<210> 5
<211> 10
<212> PRT
<213> 人工序列
<220>
<223> CDRH3
<400> 5
Asp Trp Gly Leu Arg Pro Trp Phe Val Tyr
1 5 10
<210> 6
<211> 11
<212> PRT
<213> 人工序列
<220>
<223> CDRL1
<400> 6
Lys Ala Ser Gln Asp Val Ser Thr Ala Val Ala
1 5 10
<210> 7
<211> 7
<212> PRT
<213> 人工序列
<220>
<223> CDRL2
<400> 7
Trp Ala Ser Thr Arg His Thr
1 5
<210> 8
<211> 9
<212> PRT
<213> 人工序列
<220>
<223> CDRL3
<400> 8
Gln Gln His Tyr Ser Thr Pro Pro Thr
1 5
<210> 9
<211> 119
<212> PRT
<213> 人工序列
<220>
<223> 重链可变区
<400> 9
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Tyr Met Tyr Trp Val Arg Gln Thr Pro Glu Lys Arg Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Val Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Asn Leu Tyr
65 70 75 80
Leu Gln Met Ser Ser Leu Lys Ser Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Arg Asp Trp Gly Leu Arg Pro Trp Phe Val Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ala
115
<210> 10
<211> 357
<212> DNA
<213> 人工序列
<220>
<223> 重链可变区
<400> 10
gaagtgcagc tggtggagtc tgggggaggc ttagtgaagc ctggagggtc cctgaaactc 60
tcctgtgcag cctctggatt cactttcagt gactattaca tgtattgggt tcgccagact 120
ccggaaaaga ggctggagtg ggtcgcaacc attagtgttg gtggtagttt cacctactat 180
ccagacagtg tgaaggggcg attcaccatc tccagagaca atgccaagaa caacctgtac 240
ctgcaaatga gcagtctgaa gtctgaggac acagccatgt attactgtgc aagagactgg 300
ggattacgac cctggtttgt ttactggggc caagggactc tggtcactgt ctctgca 357
<210> 11
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> 轻链可变区
<400> 11
Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly
1 5 10 15
Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Ser Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Thr Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Val Gln Ala
65 70 75 80
Glu Asp Leu Ala Leu Tyr Tyr Cys Gln Gln His Tyr Ser Thr Pro Pro
85 90 95
Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys
100 105
<210> 12
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> 轻链可变区
<400> 12
gacattgtga tgacccagtc tcacaaattc atgtccacat cagtaggaga cagggtcagc 60
atcacctgca aggccagtca ggatgtgagt actgctgtag cctggtatca acaaaaacca 120
gggcaatctc ctaaactact gatttactgg gcatccaccc ggcacactgg agtccctgat 180
cgcttcacag gcagtggatc tgggacagat tatactctca ccatcagcag tgtgcaggct 240
gaagacctgg cactttatta ctgtcagcaa cattatagca ctcctcccac gttcggctcg 300
gggacaaagt tggaaataaa a 321
<210> 13
<211> 5
<212> PRT
<213> 人工序列
<220>
<223> CDRH1
<400> 13
Ser Tyr Trp Met His
1 5
<210> 14
<211> 17
<212> PRT
<213> 人工序列
<220>
<223> CDRH2
<400> 14
Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Leu Asn Gln Lys Phe Lys
1 5 10 15
Asp
<210> 15
<211> 12
<212> PRT
<213> 人工序列
<220>
<223> CDRH3
<400> 15
Arg Phe Tyr Tyr Gly Ser Asp Trp Tyr Phe Asp Val
1 5 10
<210> 16
<211> 11
<212> PRT
<213> 人工序列
<220>
<223> CDRL1
<400> 16
Lys Ala Ser Gln Asp Val Gly Thr Ala Val Ala
1 5 10
<210> 17
<211> 7
<212> PRT
<213> 人工序列
<220>
<223> CDRL2
<400> 17
Trp Ala Ser Thr Arg His Thr
1 5
<210> 18
<211> 9
<212> PRT
<213> 人工序列
<220>
<223> CDRL3
<400> 18
Gln Gln Tyr Ser Ser Tyr Pro Leu Thr
1 5
<210> 19
<211> 121
<212> PRT
<213> 人工序列
<220>
<223> 重链可变区
<400> 19
Gln Val Gln Leu Gln Gln Ser Gly Pro Gln Leu Val Arg Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr
20 25 30
Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Leu Asn Gln Lys Phe
50 55 60
Lys Asp Lys Ala Ser Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Pro Thr Ser Gly Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Arg Phe Tyr Tyr Gly Ser Asp Trp Tyr Phe Asp Val Trp Gly
100 105 110
Ala Gly Ser Thr Val Thr Val Ser Ser
115 120
<210> 20
<211> 363
<212> DNA
<213> 人工序列
<220>
<223> 重链可变区
<400> 20
caggtgcaac tgcagcagtc tgggcctcag ctggttaggc ctggggcttc agtgaagata 60
tcctgcaagg cttctggtta ctcattcacc agctactgga tgcactgggt gaagcagagg 120
cctggacaag gtcttgagtg gattggcatg attgatcctt ccgatagtga aactaggtta 180
aatcagaagt tcaaggacaa ggcctcattg actgtagaca aatcctccag cacagcctac 240
atgcaactca gcagcccgac atctggggac tctgcggtct attactgtgc aagacgtttt 300
tactacgggt cggactggta cttcgatgtc tggggcgcag ggtccacggt caccgtctcc 360
tca 363
<210> 21
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> 轻链可变区
<400> 21
Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly
1 5 10 15
Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Thr Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser
65 70 75 80
Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln Tyr Ser Ser Tyr Pro Leu
85 90 95
Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys
100 105
<210> 22
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> 轻链可变区
<400> 22
gacattgtga tgacccagtc tcacaaattc atgtccacat cagtaggaga cagggtcagc 60
atcacctgca aggccagtca ggatgtgggt actgctgtag cctggtatca acagaaacca 120
ggtcaatctc ctaaactact gatttactgg gcatccaccc ggcacactgg agtccctgat 180
cgcttcacag gcagtggatc tgggacagat ttcactctca ccattagcaa tgtgcagtct 240
gaagacttgg cagattattt ctgtcagcaa tatagcagct atcctctcac gttcggctcg 300
gggacaaagt tggaaataaa a 321
<210> 23
<211> 5
<212> PRT
<213> 人工序列
<220>
<223> CDRH1
<400> 23
Asp Tyr Tyr Met Tyr
1 5
<210> 24
<211> 17
<212> PRT
<213> 人工序列
<220>
<223> CDRH2
<400> 24
Thr Ile Asn Asp Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val Lys
1 5 10 15
Gly
<210> 25
<211> 10
<212> PRT
<213> 人工序列
<220>
<223> CDRH3
<400> 25
Asp Trp Gly Leu Arg Pro Trp Phe Val Tyr
1 5 10
<210> 26
<211> 11
<212> PRT
<213> 人工序列
<220>
<223> CDRL1
<400> 26
Lys Ala Ser Gln Asp Val Ser Thr Ala Val Ala
1 5 10
<210> 27
<211> 7
<212> PRT
<213> 人工序列
<220>
<223> CDRL2
<400> 27
Trp Ala Ser Thr Arg His Thr
1 5
<210> 28
<211> 9
<212> PRT
<213> 人工序列
<220>
<223> CDRL3
<400> 28
Gln Gln His Tyr Thr Thr Pro Pro Thr
1 5
<210> 29
<211> 119
<212> PRT
<213> 人工序列
<220>
<223> 重链可变区
<400> 29
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Tyr Met Tyr Trp Ile Arg Gln Thr Pro Glu Lys Arg Leu Glu Trp Val
35 40 45
Ala Thr Ile Asn Asp Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Asn Leu Tyr
65 70 75 80
Leu Gln Met Ser Ser Leu Lys Ser Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Arg Asp Trp Gly Leu Arg Pro Trp Phe Val Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ala
115
<210> 30
<211> 357
<212> DNA
<213> 人工序列
<220>
<223> 重链可变区
<400> 30
gaagtgcagc tggtggagtc tgggggaggc ttagtgaagc ctggagggtc cctgaaactc 60
tcctgtgcag cctctggatt cactttcagt gactattaca tgtattggat tcgccagact 120
ccggaaaaga ggctggagtg ggtcgcaacc attaatgatg gtggtagtta cacctactat 180
ccagacagtg tgaaggggcg attcaccatc tccagagaca atgccaagaa caacctgtac 240
ctgcaaatga gcagtctgaa gtctgaggac acagccatgt attactgtgc aagagactgg 300
ggattacgac cctggtttgt ttactggggc caagggactc tggtcactgt ctctgca 357
<210> 31
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> 轻链可变区
<400> 31
Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly
1 5 10 15
Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Ser Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Thr Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Val Gln Ala
65 70 75 80
Glu Asp Leu Ala Leu Tyr Tyr Cys Gln Gln His Tyr Thr Thr Pro Pro
85 90 95
Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys
100 105
<210> 32
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> 轻链可变区
<400> 32
gacattgtga tgacccagtc tcacaaattc atgtccacat cagtaggaga cagggtcagc 60
atcacctgca aggccagtca ggatgtgagt actgctgtag cctggtatca acaaaaacca 120
gggcaatctc ctaaactact gatttactgg gcatccaccc ggcacactgg agtccctgat 180
cgcttcacag gcagtggatc tgggacagat tatactctca ccatcagcag tgtgcaggct 240
gaagacctgg cactttatta ctgtcagcaa cattatacca ctcctcccac gttcggctcg 300
gggacaaagt tggaaataaa a 321
<210> 33
<211> 5
<212> PRT
<213> 人工序列
<220>
<223> CDRH1
<400> 33
Gly Tyr Asn Met Asn
1 5
<210> 34
<211> 17
<212> PRT
<213> 人工序列
<220>
<223> CDRH2
<400> 34
Asn Ile Asp Pro Tyr Tyr Gly Gly Thr Ser Tyr Asn Gln Lys Phe Lys
1 5 10 15
Gly
<210> 35
<211> 7
<212> PRT
<213> 人工序列
<220>
<223> CDRH3
<400> 35
Tyr Gly Asn Tyr Val Asp Tyr
1 5
<210> 36
<211> 11
<212> PRT
<213> 人工序列
<220>
<223> CDRL1
<400> 36
Lys Ala Ser Gln Asp Val Ser Thr Ala Val Ala
1 5 10
<210> 37
<211> 7
<212> PRT
<213> 人工序列
<220>
<223> CDRL2
<400> 37
Trp Ala Ser Thr Arg His Thr
1 5
<210> 38
<211> 9
<212> PRT
<213> 人工序列
<220>
<223> CDRL3
<400> 38
Gln Gln His Tyr Asn Thr Pro Pro Thr
1 5
<210> 39
<211> 116
<212> PRT
<213> 人工序列
<220>
<223> 重链可变区
<400> 39
Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Glu Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Gly Tyr
20 25 30
Asn Met Asn Trp Val Lys Gln Ser Asn Gly Lys Ser Leu Glu Trp Ile
35 40 45
Gly Asn Ile Asp Pro Tyr Tyr Gly Gly Thr Ser Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Lys Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Val Arg Tyr Gly Asn Tyr Val Asp Tyr Trp Gly Gln Gly Thr Thr Leu
100 105 110
Thr Val Ser Ser
115
<210> 40
<211> 342
<212> DNA
<213> 人工序列
<220>
<223> 重链可变区
<400> 40
cagctgcagc agtctggacc tgagctggag aagcctggcg cttcagtgaa gatatcctgc 60
aaggcttctg gttactcatt cactggctac aacatgaact gggtgaagca gagcaatgga 120
aagagccttg agtggattgg aaatattgat ccttactatg gtggtactag ctacaaccag 180
aagttcaagg gcaaggccac attgactgta gacaaatcct ccagcacagc ctacatgcag 240
ctcaagagcc tgacatctga ggactctgca gtctattact gtgtaaggta tggtaactac 300
gtggactact ggggccaagg caccactctc acagtctcct ca 342
<210> 41
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> 轻链可变区
<400> 41
Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly
1 5 10 15
Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Ser Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Thr Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Val Gln Ala
65 70 75 80
Glu Asp Leu Ala Leu Tyr Tyr Cys Gln Gln His Tyr Asn Thr Pro Pro
85 90 95
Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys
100 105
<210> 42
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> 轻链可变区
<400> 42
gacattgtga tgacccagtc ccacaaattc atgtccacat cagtaggaga cagggtcagc 60
atcacctgca aggccagtca ggatgtgagt actgctgtag cctggtatca acaaaaacca 120
gggcaatctc ctaaactact gatttactgg gcatccaccc ggcacactgg agtccctgat 180
cgcttcacag gcagtggatc tgggacagat tatactctca ccatcagcag tgtgcaggct 240
gaagacctgg cactttatta ctgtcagcaa cattataaca ctcctcccac gttcggctcg 300
gggacaaagt tggaaataaa a 321
<210> 43
<211> 119
<212> PRT
<213> 人工序列
<220>
<223> VH
<400> 43
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Tyr Met Tyr Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Thr Ile Ser Val Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Trp Gly Leu Arg Pro Trp Phe Val Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser
115
<210> 44
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> VL
<400> 44
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Ser Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Ser Thr Pro Pro
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 45
<211> 357
<212> DNA
<213> 人工序列
<220>
<223> VH
<400> 45
caggtacagc tcgtggagtc tggtggaggc ttggtgaaac ctggagggtc cctgagactt 60
agctgtgcag cttccggctt cacattttca gactattata tgtattggat cagacaggct 120
cccgggaagg gcttggagtg ggtttcaacc attagtgttg gcggatcttt tacttactac 180
ccagacagtg tgaaggggag attcacaatc tccagggata acgcgaaaaa cagcctgtat 240
ctccaaatga atagcctgag agccgaagat accgccgtgt actactgcgc cagagactgg 300
ggattacggc cctggttcgt gtactggggc cagggaaccc tggtcaccgt ctcctca 357
<210> 46
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> VL
<400> 46
gacatccaga tgacacagtc cccaagctcc ctgtctgcat ctgtgggaga ccgggtgacc 60
atcacttgta aggcctcaca ggatgtttct actgctgtcg catggtacca gcaaaagccg 120
ggtaaagctc ccaagctttt gatatactgg gccagcacca ggcacacagg cgtgccatca 180
agattcagtg ggtccggatc cggcacggat tttacactca ctattagctc actgcaacct 240
gaagactttg ccacctatta ctgccagcag cattatagca cccctcccac cttcggtcag 300
ggcactaaag tagaaatcaa a 321
<210> 47
<211> 119
<212> PRT
<213> 人工序列
<220>
<223> VH
<400> 47
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Thr Ile Ser Val Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Trp Gly Leu Arg Pro Trp Phe Val Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser
115
<210> 48
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> VL
<400> 48
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Ser Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Ser Thr Pro Pro
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 49
<211> 357
<212> DNA
<213> 人工序列
<220>
<223> VH
<400> 49
caggtccagc tggtggaatc cggcggaggc ttggtgaagc ctggaggcag cctaagactc 60
tcctgtgcag cctctggctt caccttctct gactattaca tgtattgggt ccgccaggct 120
ccagggaaag ggctcgagtg ggtttcaaca attagtgtag gtggaagctt cacctactat 180
cctgactccg tgaaaggaag atttacgatc tctagggata atgccaagaa ctcactgtac 240
cttcagatga acagcctgag agcggaggac acagccgtgt actactgcgc tagagattgg 300
ggattaagac cctggtttgt ttattggggc cagggaaccc tggtcaccgt ctcctca 357
<210> 50
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> VL
<400> 50
gacatccaga tgacacagtc cccaagctcc ctgtctgcat ctgtgggaga ccgggtgacc 60
atcacttgta aggcctcaca ggatgtttct actgctgtcg catggtacca gcaaaagccg 120
ggtaaagctc ccaagctttt gatatactgg gccagcacca ggcacacagg cgtgccatca 180
agattcagtg ggtccggatc cggcacggat tttacactca ctattagctc actgcaacct 240
gaagactttg ccacctatta ctgccagcag cattatagca cccctcccac cttcggtcag 300
ggcactaaag tagaaatcaa a 321
<210> 51
<211> 119
<212> PRT
<213> 人工序列
<220>
<223> VH
<400> 51
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Val Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Trp Gly Leu Arg Pro Trp Phe Val Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser
115
<210> 52
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> VL
<400> 52
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Ser Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Ser Thr Pro Pro
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 53
<211> 357
<212> DNA
<213> 人工序列
<220>
<223> VH
<400> 53
caggtgcagc tggtcgaatc tggaggaggc ttggtgaaac ctggggggtc cctgagactc 60
tcttgtgcag cctccggctt taccttttct gactactaca tgtattgggt tcgccaggct 120
cccggtaagg ggttagagtg ggtggctacc attagtgttg gcggttcatt tacttattac 180
ccagatagtg tgaaaggacg gttcaccatc agcagggaca atgcaaagaa ctcactctat 240
ctacaaatga atagcctgag agccgaggat acagcggtgt attactgcgc cagagattgg 300
ggacttcgac catggttcgt ctactggggc cagggaaccc tggtcaccgt ctcctca 357
<210> 54
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> VL
<400> 54
gacatccaga tgacacagtc cccaagctcc ctgtctgcat ctgtgggaga ccgggtgacc 60
atcacttgta aggcctcaca ggatgtttct actgctgtcg catggtacca gcaaaagccg 120
ggtaaagctc ccaagctttt gatatactgg gccagcacca ggcacacagg cgtgccatca 180
agattcagtg ggtccggatc cggcacggat tttacactca ctattagctc actgcaacct 240
gaagactttg ccacctatta ctgccagcag cattatagca cccctcccac cttcggtcag 300
ggcactaaag tagaaatcaa a 321
<210> 55
<211> 121
<212> PRT
<213> 人工序列
<220>
<223> VH
<400> 55
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr
20 25 30
Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Leu Asn Gln Lys Phe
50 55 60
Lys Asp Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Arg Phe Tyr Tyr Gly Ser Asp Trp Tyr Phe Asp Val Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 56
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> VL
<400> 56
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Ser Tyr Pro Leu
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 57
<211> 363
<212> DNA
<213> 人工序列
<220>
<223> VH
<400> 57
caggtgcagc tggtgcagag tggagctgag gtaaaaaagc ccggcgccag tgtgaaggtt 60
agttgcaagg cctctggata caccttcaca agctattgga tgcactgggt gcgacaagct 120
cctgggcagg ggcttgagtg gatgggaatg atcgacccat ccgattcaga aactaggctc 180
aaccagaaat tcaaagatag agtgactatg accagggaca cctccacgag cacagtctac 240
atggaattgt caagcctgcg ctctgaggac acagccgtgt actattgtgc aagacggttt 300
tactatggta gcgattggta ctttgatgtt tggggccagg gaaccctggt caccgtctcc 360
tca 363
<210> 58
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> VL
<400> 58
gacatacagt tgacccagtc tccttccttc ctgtccgcct ccgtgggcga tagagttacc 60
attacttgca aagctagtca ggacgtgggt accgcagtgg cctggtatca gcagaaacca 120
ggtaaagccc ctaagctcct gatctactgg gcatcaacac ggcacacagg ggtcccaagc 180
aggttttctg gcagcggatc aggaaccgaa tttacactga cgatctcgtc tctgcagccc 240
gaggatttcg ctacttacta ctgtcaacaa tatagtagct atcccctcac tttcggtcag 300
ggcactaaag tagaaatcaa a 321
<210> 59
<211> 121
<212> PRT
<213> 人工序列
<220>
<223> VH
<400> 59
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr
20 25 30
Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Leu Asn Gln Lys Phe
50 55 60
Lys Asp Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Arg Phe Tyr Tyr Gly Ser Asp Trp Tyr Phe Asp Val Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 60
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> VL
<400> 60
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Ser Tyr Pro Leu
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 61
<211> 363
<212> DNA
<213> 人工序列
<220>
<223> VH
<400> 61
caggtgcaac tcgtgcagtc tggagctgaa gtgaagaaac ccggggcctc agtgaaggtg 60
agttgcaaag catctgggta ctcatttacc agctattgga tgcactgggt gcggcaggcc 120
ccaggacaag gcctggagtg gattggcatg atcgaccctt ccgatagtga aacgaggctg 180
aaccagaagt ttaaagatcg cgtcaccatg accagggaca caagtacttc tacagtctac 240
atggagttga gcagcctgag atcagaggac acagccgttt actactgtgc tagacgattc 300
tattatggca gcgactggta tttcgatgta tggggccagg gaaccctggt caccgtctcc 360
tca 363
<210> 62
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> VL
<400> 62
gacatacagt tgacccagtc tccttccttc ctgtccgcct ccgtgggcga tagagttacc 60
attacttgca aagctagtca ggacgtgggt accgcagtgg cctggtatca gcagaaacca 120
ggtaaagccc ctaagctcct gatctactgg gcatcaacac ggcacacagg ggtcccaagc 180
aggttttctg gcagcggatc aggaaccgaa tttacactga cgatctcgtc tctgcagccc 240
gaggatttcg ctacttacta ctgtcaacaa tatagtagct atcccctcac tttcggtcag 300
ggcactaaag tagaaatcaa a 321
<210> 63
<211> 121
<212> PRT
<213> 人工序列
<220>
<223> VH
<400> 63
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr
20 25 30
Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Leu Asn Gln Lys Phe
50 55 60
Lys Asp Lys Ala Ser Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Arg Phe Tyr Tyr Gly Ser Asp Trp Tyr Phe Asp Val Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 64
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> VL
<400> 64
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Ser Tyr Pro Leu
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 65
<211> 363
<212> DNA
<213> 人工序列
<220>
<223> VH
<400> 65
caggtgcaac tggtgcagtc tggtgctgag gtgaagaaac caggcgcttc agtcaaggta 60
agctgcaaag caagtggata ctccttcacc tcttattgga tgcactgggt tagacaggcc 120
cctggtcaag gcctcgagtg gattggcatg atcgacccct ctgacagcga aactaggctg 180
aatcagaaat ttaaggacaa ggcctccatg acacgggata catccacaag caccgtttac 240
atggaactga gctcgctgag aagtgaggac actgccgtgt attactgtgc gagacgcttt 300
tattacgggt cagattggta cttcgatgtg tggggccagg gaaccctggt caccgtctcc 360
tca 363
<210> 66
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> VL
<400> 66
gacatacagt tgacccagtc tccttccttc ctgtccgcct ccgtgggcga tagagttacc 60
attacttgca aagctagtca ggacgtgggt accgcagtgg cctggtatca gcagaaacca 120
ggtaaagccc ctaagctcct gatctactgg gcatcaacac ggcacacagg ggtcccaagc 180
aggttttctg gcagcggatc aggaaccgaa tttacactga cgatctcgtc tctgcagccc 240
gaggatttcg ctacttacta ctgtcaacaa tatagtagct atcccctcac tttcggtcag 300
ggcactaaag tagaaatcaa a 321
<210> 67
<211> 121
<212> PRT
<213> 人工序列
<220>
<223> VH
<400> 67
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr
20 25 30
Trp Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Leu Asn Gln Lys Phe
50 55 60
Lys Asp Lys Ala Ser Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Arg Phe Tyr Tyr Gly Ser Asp Trp Tyr Phe Asp Val Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 68
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> VL
<400> 68
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Ser Tyr Pro Leu
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 69
<211> 363
<212> DNA
<213> 人工序列
<220>
<223> VH
<400> 69
caggtgcagc tggtgcagtc tggggctgag gtgaaaaagc caggcgcttc cgtcaaagtt 60
tcctgcaagg catctggtta ctcttttaca agctattgga tgcactgggt gaagcaggcc 120
cccggacaag ggctcgagtg gattggcatg atcgatcctt ccgatagtga aacacgcttg 180
aatcagaaat tcaaggacaa ggccagtatg accagggata ctagcacaag cactgtatat 240
atggagctta gctcactgag atcagaagac acggccgtgt actactgtgc gagacggttt 300
tactatggct ccgactggta tttcgacgtc tggggccagg gaaccctggt caccgtctcc 360
tca 363
<210> 70
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> VL
<400> 70
gacatacagt tgacccagtc tccttccttc ctgtccgcct ccgtgggcga tagagttacc 60
attacttgca aagctagtca ggacgtgggt accgcagtgg cctggtatca gcagaaacca 120
ggtaaagccc ctaagctcct gatctactgg gcatcaacac ggcacacagg ggtcccaagc 180
aggttttctg gcagcggatc aggaaccgaa tttacactga cgatctcgtc tctgcagccc 240
gaggatttcg ctacttacta ctgtcaacaa tatagtagct atcccctcac tttcggtcag 300
ggcactaaag tagaaatcaa a 321
<210> 71
<211> 121
<212> PRT
<213> 人工序列
<220>
<223> VH
<400> 71
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr
20 25 30
Trp Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Leu Asn Gln Lys Phe
50 55 60
Lys Asp Lys Ala Ser Leu Thr Val Asp Thr Ser Thr Ser Thr Val Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Arg Phe Tyr Tyr Gly Ser Asp Trp Tyr Phe Asp Val Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 72
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> VL
<400> 72
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Ser Tyr Pro Leu
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 73
<211> 363
<212> DNA
<213> 人工序列
<220>
<223> VH
<400> 73
caggtgcagc tggtgcagtc tggcgctgag gtgaagaaac ctggggcctc agtgaaggtt 60
tcctgtaaag caagtggata ctctttcacc agctactgga tgcactgggt gaaacaggcc 120
cccggccaag ggcttgagtg gattggtatg atcgatccat ccgacagcga aactaggctc 180
aaccagaagt tcaaggataa agcgtccttg acagtagata catccacgag cacagtttat 240
atggagctgt ctagtctgcg gtctgaagac accgccgtgt attattgcgc tagacgcttt 300
tattacggct cggactggta ctttgacgtc tggggccagg gaaccctggt caccgtctcc 360
tca 363
<210> 74
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> VL
<400> 74
gacatacagt tgacccagtc tccttccttc ctgtccgcct ccgtgggcga tagagttacc 60
attacttgca aagctagtca ggacgtgggt accgcagtgg cctggtatca gcagaaacca 120
ggtaaagccc ctaagctcct gatctactgg gcatcaacac ggcacacagg ggtcccaagc 180
aggttttctg gcagcggatc aggaaccgaa tttacactga cgatctcgtc tctgcagccc 240
gaggatttcg ctacttacta ctgtcaacaa tatagtagct atcccctcac tttcggtcag 300
ggcactaaag tagaaatcaa a 321
<210> 75
<211> 121
<212> PRT
<213> 人工序列
<220>
<223> VH
<400> 75
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr
20 25 30
Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Leu Asn Gln Lys Phe
50 55 60
Lys Asp Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Arg Phe Tyr Tyr Gly Ser Asp Trp Tyr Phe Asp Val Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 76
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> VL
<400> 76
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Ser Tyr Pro Leu
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 77
<211> 363
<212> DNA
<213> 人工序列
<220>
<223> VH
<400> 77
caggtgcagc tggtgcagag tggagctgag gtaaaaaagc ccggcgccag tgtgaaggtt 60
agttgcaagg cctctggata caccttcaca agctattgga tgcactgggt gcgacaagct 120
cctgggcagg ggcttgagtg gatgggaatg atcgacccat ccgattcaga aactaggctc 180
aaccagaaat tcaaagatag agtgactatg accagggaca cctccacgag cacagtctac 240
atggaattgt caagcctgcg ctctgaggac acagccgtgt actattgtgc aagacggttt 300
tactatggta gcgattggta ctttgatgtt tggggccagg gaaccctggt caccgtctcc 360
tca 363
<210> 78
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> VL
<400> 78
gatattcaac tcacccagag tccatccttc ctgtctgcct cagtgggcga cagagtgtca 60
atcacatgca aggcaagcca ggatgttggc actgctgtgg cttggtatca gcaaaaacca 120
ggtaaggccc ccaaactgct tatttactgg gcatcaaccc ggcacacggg tgtccccgac 180
aggttcagcg gcagtggatc tgggacagag tttaccctga ctatcagctc cctgcagcct 240
gaagactttg ccacttatta ctgtcagcag tactctagct atcctctcac cttcggtcag 300
ggcactaaag tagaaatcaa a 321
<210> 79
<211> 121
<212> PRT
<213> 人工序列
<220>
<223> VH
<400> 79
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr
20 25 30
Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Leu Asn Gln Lys Phe
50 55 60
Lys Asp Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Arg Phe Tyr Tyr Gly Ser Asp Trp Tyr Phe Asp Val Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 80
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> VL
<400> 80
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Ser Tyr Pro Leu
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 81
<211> 363
<212> DNA
<213> 人工序列
<220>
<223> VH
<400> 81
caggtgcaac tcgtgcagtc tggagctgaa gtgaagaaac ccggggcctc agtgaaggtg 60
agttgcaaag catctgggta ctcatttacc agctattgga tgcactgggt gcggcaggcc 120
ccaggacaag gcctggagtg gattggcatg atcgaccctt ccgatagtga aacgaggctg 180
aaccagaagt ttaaagatcg cgtcaccatg accagggaca caagtacttc tacagtctac 240
atggagttga gcagcctgag atcagaggac acagccgttt actactgtgc tagacgattc 300
tattatggca gcgactggta tttcgatgta tggggccagg gaaccctggt caccgtctcc 360
tca 363
<210> 82
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> VL
<400> 82
gatattcaac tcacccagag tccatccttc ctgtctgcct cagtgggcga cagagtgtca 60
atcacatgca aggcaagcca ggatgttggc actgctgtgg cttggtatca gcaaaaacca 120
ggtaaggccc ccaaactgct tatttactgg gcatcaaccc ggcacacggg tgtccccgac 180
aggttcagcg gcagtggatc tgggacagag tttaccctga ctatcagctc cctgcagcct 240
gaagactttg ccacttatta ctgtcagcag tactctagct atcctctcac cttcggtcag 300
ggcactaaag tagaaatcaa a 321
<210> 83
<211> 121
<212> PRT
<213> 人工序列
<220>
<223> VH
<400> 83
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr
20 25 30
Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Leu Asn Gln Lys Phe
50 55 60
Lys Asp Lys Ala Ser Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Arg Phe Tyr Tyr Gly Ser Asp Trp Tyr Phe Asp Val Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 84
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> VL
<400> 84
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Ser Tyr Pro Leu
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 85
<211> 363
<212> DNA
<213> 人工序列
<220>
<223> VH
<400> 85
caggtgcaac tggtgcagtc tggtgctgag gtgaagaaac caggcgcttc agtcaaggta 60
agctgcaaag caagtggata ctccttcacc tcttattgga tgcactgggt tagacaggcc 120
cctggtcaag gcctcgagtg gattggcatg atcgacccct ctgacagcga aactaggctg 180
aatcagaaat ttaaggacaa ggcctccatg acacgggata catccacaag caccgtttac 240
atggaactga gctcgctgag aagtgaggac actgccgtgt attactgtgc gagacgcttt 300
tattacgggt cagattggta cttcgatgtg tggggccagg gaaccctggt caccgtctcc 360
tca 363
<210> 86
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> VL
<400> 86
gatattcaac tcacccagag tccatccttc ctgtctgcct cagtgggcga cagagtgtca 60
atcacatgca aggcaagcca ggatgttggc actgctgtgg cttggtatca gcaaaaacca 120
ggtaaggccc ccaaactgct tatttactgg gcatcaaccc ggcacacggg tgtccccgac 180
aggttcagcg gcagtggatc tgggacagag tttaccctga ctatcagctc cctgcagcct 240
gaagactttg ccacttatta ctgtcagcag tactctagct atcctctcac cttcggtcag 300
ggcactaaag tagaaatcaa a 321
<210> 87
<211> 121
<212> PRT
<213> 人工序列
<220>
<223> VH
<400> 87
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr
20 25 30
Trp Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Leu Asn Gln Lys Phe
50 55 60
Lys Asp Lys Ala Ser Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Arg Phe Tyr Tyr Gly Ser Asp Trp Tyr Phe Asp Val Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 88
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> VL
<400> 88
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Ser Tyr Pro Leu
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 89
<211> 363
<212> DNA
<213> 人工序列
<220>
<223> VH
<400> 89
caggtgcagc tggtgcagtc tggggctgag gtgaaaaagc caggcgcttc cgtcaaagtt 60
tcctgcaagg catctggtta ctcttttaca agctattgga tgcactgggt gaagcaggcc 120
cccggacaag ggctcgagtg gattggcatg atcgatcctt ccgatagtga aacacgcttg 180
aatcagaaat tcaaggacaa ggccagtatg accagggata ctagcacaag cactgtatat 240
atggagctta gctcactgag atcagaagac acggccgtgt actactgtgc gagacggttt 300
tactatggct ccgactggta tttcgacgtc tggggccagg gaaccctggt caccgtctcc 360
tca 363
<210> 90
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> VL
<400> 90
gatattcaac tcacccagag tccatccttc ctgtctgcct cagtgggcga cagagtgtca 60
atcacatgca aggcaagcca ggatgttggc actgctgtgg cttggtatca gcaaaaacca 120
ggtaaggccc ccaaactgct tatttactgg gcatcaaccc ggcacacggg tgtccccgac 180
aggttcagcg gcagtggatc tgggacagag tttaccctga ctatcagctc cctgcagcct 240
gaagactttg ccacttatta ctgtcagcag tactctagct atcctctcac cttcggtcag 300
ggcactaaag tagaaatcaa a 321
<210> 91
<211> 121
<212> PRT
<213> 人工序列
<220>
<223> VH
<400> 91
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr
20 25 30
Trp Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Leu Asn Gln Lys Phe
50 55 60
Lys Asp Lys Ala Ser Leu Thr Val Asp Thr Ser Thr Ser Thr Val Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Arg Phe Tyr Tyr Gly Ser Asp Trp Tyr Phe Asp Val Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 92
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> VL
<400> 92
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Ser Tyr Pro Leu
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 93
<211> 363
<212> DNA
<213> 人工序列
<220>
<223> VH
<400> 93
caggtgcagc tggtgcagtc tggcgctgag gtgaagaaac ctggggcctc agtgaaggtt 60
tcctgtaaag caagtggata ctctttcacc agctactgga tgcactgggt gaaacaggcc 120
cccggccaag ggcttgagtg gattggtatg atcgatccat ccgacagcga aactaggctc 180
aaccagaagt tcaaggataa agcgtccttg acagtagata catccacgag cacagtttat 240
atggagctgt ctagtctgcg gtctgaagac accgccgtgt attattgcgc tagacgcttt 300
tattacggct cggactggta ctttgacgtc tggggccagg gaaccctggt caccgtctcc 360
tca 363
<210> 94
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> VL
<400> 94
gatattcaac tcacccagag tccatccttc ctgtctgcct cagtgggcga cagagtgtca 60
atcacatgca aggcaagcca ggatgttggc actgctgtgg cttggtatca gcaaaaacca 120
ggtaaggccc ccaaactgct tatttactgg gcatcaaccc ggcacacggg tgtccccgac 180
aggttcagcg gcagtggatc tgggacagag tttaccctga ctatcagctc cctgcagcct 240
gaagactttg ccacttatta ctgtcagcag tactctagct atcctctcac cttcggtcag 300
ggcactaaag tagaaatcaa a 321
<210> 95
<211> 121
<212> PRT
<213> 人工序列
<220>
<223> VH
<400> 95
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr
20 25 30
Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Leu Asn Gln Lys Phe
50 55 60
Lys Asp Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Arg Phe Tyr Tyr Gly Ser Asp Trp Tyr Phe Asp Val Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 96
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> VL
<400> 96
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Asp Tyr Phe Cys Gln Gln Tyr Ser Ser Tyr Pro Leu
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 97
<211> 363
<212> DNA
<213> 人工序列
<220>
<223> VH
<400> 97
caggtgcagc tggtgcagag tggagctgag gtaaaaaagc ccggcgccag tgtgaaggtt 60
agttgcaagg cctctggata caccttcaca agctattgga tgcactgggt gcgacaagct 120
cctgggcagg ggcttgagtg gatgggaatg atcgacccat ccgattcaga aactaggctc 180
aaccagaaat tcaaagatag agtgactatg accagggaca cctccacgag cacagtctac 240
atggaattgt caagcctgcg ctctgaggac acagccgtgt actattgtgc aagacggttt 300
tactatggta gcgattggta ctttgatgtt tggggccagg gaaccctggt caccgtctcc 360
tca 363
<210> 98
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> VL
<400> 98
gacatccagt tgacccaatc accatccttt ctgtctgcct ctgtgggaga tagagtctcc 60
attacttgca aggccagtca ggatgtgggg accgctgttg cctggtacca gcaaaaaccc 120
ggaaaggcac ctaaactcct tatctactgg gcatccaccc ggcacacagg agtgccagac 180
aggtttagcg ggtcaggctc tggtacagag ttcactctga caatttctag cctgcagcct 240
gaagacttcg ctgattattt ctgtcagcag tatagcagtt accccctcac gttcggtcag 300
ggcactaaag tagaaatcaa a 321
<210> 99
<211> 121
<212> PRT
<213> 人工序列
<220>
<223> VH
<400> 99
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr
20 25 30
Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Leu Asn Gln Lys Phe
50 55 60
Lys Asp Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Arg Phe Tyr Tyr Gly Ser Asp Trp Tyr Phe Asp Val Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 100
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> VL
<400> 100
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Asp Tyr Phe Cys Gln Gln Tyr Ser Ser Tyr Pro Leu
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 101
<211> 363
<212> DNA
<213> 人工序列
<220>
<223> VH
<400> 101
caggtgcaac tcgtgcagtc tggagctgaa gtgaagaaac ccggggcctc agtgaaggtg 60
agttgcaaag catctgggta ctcatttacc agctattgga tgcactgggt gcggcaggcc 120
ccaggacaag gcctggagtg gattggcatg atcgaccctt ccgatagtga aacgaggctg 180
aaccagaagt ttaaagatcg cgtcaccatg accagggaca caagtacttc tacagtctac 240
atggagttga gcagcctgag atcagaggac acagccgttt actactgtgc tagacgattc 300
tattatggca gcgactggta tttcgatgta tggggccagg gaaccctggt caccgtctcc 360
tca 363
<210> 102
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> VL
<400> 102
gacatccagt tgacccaatc accatccttt ctgtctgcct ctgtgggaga tagagtctcc 60
attacttgca aggccagtca ggatgtgggg accgctgttg cctggtacca gcaaaaaccc 120
ggaaaggcac ctaaactcct tatctactgg gcatccaccc ggcacacagg agtgccagac 180
aggtttagcg ggtcaggctc tggtacagag ttcactctga caatttctag cctgcagcct 240
gaagacttcg ctgattattt ctgtcagcag tatagcagtt accccctcac gttcggtcag 300
ggcactaaag tagaaatcaa a 321
<210> 103
<211> 121
<212> PRT
<213> 人工序列
<220>
<223> VH
<400> 103
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr
20 25 30
Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Leu Asn Gln Lys Phe
50 55 60
Lys Asp Lys Ala Ser Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Arg Phe Tyr Tyr Gly Ser Asp Trp Tyr Phe Asp Val Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 104
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> VL
<400> 104
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Asp Tyr Phe Cys Gln Gln Tyr Ser Ser Tyr Pro Leu
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 105
<211> 363
<212> DNA
<213> 人工序列
<220>
<223> VH
<400> 105
caggtgcaac tggtgcagtc tggtgctgag gtgaagaaac caggcgcttc agtcaaggta 60
agctgcaaag caagtggata ctccttcacc tcttattgga tgcactgggt tagacaggcc 120
cctggtcaag gcctcgagtg gattggcatg atcgacccct ctgacagcga aactaggctg 180
aatcagaaat ttaaggacaa ggcctccatg acacgggata catccacaag caccgtttac 240
atggaactga gctcgctgag aagtgaggac actgccgtgt attactgtgc gagacgcttt 300
tattacgggt cagattggta cttcgatgtg tggggccagg gaaccctggt caccgtctcc 360
tca 363
<210> 106
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> VL
<400> 106
gacatccagt tgacccaatc accatccttt ctgtctgcct ctgtgggaga tagagtctcc 60
attacttgca aggccagtca ggatgtgggg accgctgttg cctggtacca gcaaaaaccc 120
ggaaaggcac ctaaactcct tatctactgg gcatccaccc ggcacacagg agtgccagac 180
aggtttagcg ggtcaggctc tggtacagag ttcactctga caatttctag cctgcagcct 240
gaagacttcg ctgattattt ctgtcagcag tatagcagtt accccctcac gttcggtcag 300
ggcactaaag tagaaatcaa a 321
<210> 107
<211> 121
<212> PRT
<213> 人工序列
<220>
<223> VH
<400> 107
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr
20 25 30
Trp Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Leu Asn Gln Lys Phe
50 55 60
Lys Asp Lys Ala Ser Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Arg Phe Tyr Tyr Gly Ser Asp Trp Tyr Phe Asp Val Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 108
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> VL
<400> 108
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Asp Tyr Phe Cys Gln Gln Tyr Ser Ser Tyr Pro Leu
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 109
<211> 363
<212> DNA
<213> 人工序列
<220>
<223> VH
<400> 109
caggtgcagc tggtgcagtc tggggctgag gtgaaaaagc caggcgcttc cgtcaaagtt 60
tcctgcaagg catctggtta ctcttttaca agctattgga tgcactgggt gaagcaggcc 120
cccggacaag ggctcgagtg gattggcatg atcgatcctt ccgatagtga aacacgcttg 180
aatcagaaat tcaaggacaa ggccagtatg accagggata ctagcacaag cactgtatat 240
atggagctta gctcactgag atcagaagac acggccgtgt actactgtgc gagacggttt 300
tactatggct ccgactggta tttcgacgtc tggggccagg gaaccctggt caccgtctcc 360
tca 363
<210> 110
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> VL
<400> 110
gacatccagt tgacccaatc accatccttt ctgtctgcct ctgtgggaga tagagtctcc 60
attacttgca aggccagtca ggatgtgggg accgctgttg cctggtacca gcaaaaaccc 120
ggaaaggcac ctaaactcct tatctactgg gcatccaccc ggcacacagg agtgccagac 180
aggtttagcg ggtcaggctc tggtacagag ttcactctga caatttctag cctgcagcct 240
gaagacttcg ctgattattt ctgtcagcag tatagcagtt accccctcac gttcggtcag 300
ggcactaaag tagaaatcaa a 321
<210> 111
<211> 121
<212> PRT
<213> 人工序列
<220>
<223> VH
<400> 111
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr
20 25 30
Trp Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Leu Asn Gln Lys Phe
50 55 60
Lys Asp Lys Ala Ser Leu Thr Val Asp Thr Ser Thr Ser Thr Val Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Arg Phe Tyr Tyr Gly Ser Asp Trp Tyr Phe Asp Val Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 112
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> VL
<400> 112
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Asp Tyr Phe Cys Gln Gln Tyr Ser Ser Tyr Pro Leu
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 113
<211> 363
<212> DNA
<213> 人工序列
<220>
<223> VH
<400> 113
caggtgcagc tggtgcagtc tggcgctgag gtgaagaaac ctggggcctc agtgaaggtt 60
tcctgtaaag caagtggata ctctttcacc agctactgga tgcactgggt gaaacaggcc 120
cccggccaag ggcttgagtg gattggtatg atcgatccat ccgacagcga aactaggctc 180
aaccagaagt tcaaggataa agcgtccttg acagtagata catccacgag cacagtttat 240
atggagctgt ctagtctgcg gtctgaagac accgccgtgt attattgcgc tagacgcttt 300
tattacggct cggactggta ctttgacgtc tggggccagg gaaccctggt caccgtctcc 360
tca 363
<210> 114
<211> 321
<212> DNA
<213> 人工序列
<220>
<223> VL
<400> 114
gacatccagt tgacccaatc accatccttt ctgtctgcct ctgtgggaga tagagtctcc 60
attacttgca aggccagtca ggatgtgggg accgctgttg cctggtacca gcaaaaaccc 120
ggaaaggcac ctaaactcct tatctactgg gcatccaccc ggcacacagg agtgccagac 180
aggtttagcg ggtcaggctc tggtacagag ttcactctga caatttctag cctgcagcct 240
gaagacttcg ctgattattt ctgtcagcag tatagcagtt accccctcac gttcggtcag 300
ggcactaaag tagaaatcaa a 321
<210> 115
<211> 18
<212> PRT
<213> 人工序列
<220>
<223> 表位
<400> 115
Gln Arg Thr Trp Lys Glu Tyr Lys Val Gly Phe Gly Asn Pro Ser Gly
1 5 10 15
Glu Tyr
<210> 116
<211> 11
<212> PRT
<213> 人工序列
<220>
<223> 表位
<400> 116
Lys Ser Gly His Thr Thr Asn Gly Ile Tyr Thr
1 5 10
<210> 117
<211> 7
<212> PRT
<213> 人工序列
<220>
<223> 表位
<400> 117
Glu Ala Gly Gly Gly Gly Trp
1 5
Claims (26)
1.一种特异性地结合人血管生成素-2并诱导Tie2激活的抗体或其抗原结合片段,其中所述抗体或其抗原结合片段与SEQ ID NO:115的氨基酸、SEQ ID NO:116的氨基酸或SEQ IDNO:117的氨基酸结合。
2.根据权利要求1所述的抗体或其抗原结合片段,其中所述抗体或其抗原结合片段与SEQ ID NO:116的氨基酸或SEQ ID NO:117的氨基酸结合。
3.根据权利要求1所述的抗体或其抗原结合片段,其中所述抗体或其抗原结合片段与SEQ ID NO:115的氨基酸结合。
4.根据权利要求1所述的抗体或其抗原结合片段,其中所述抗体或其抗原结合片段与人和小鼠Ang2结合。
5.根据权利要求1所述的抗体或其抗原结合片段,其中所述抗体是多克隆的或单克隆的。
6.根据权利要求1所述的抗体或其抗原结合片段,其中所述抗原结合片段是scFv或Fab。
7.根据权利要求1所述的抗体或其抗原结合片段,其中所述抗体是人源化的。
8.根据权利要求1所述的抗体或其抗原结合片段,所述抗体或其抗原结合片段包含:
(a)具有SEQ ID NO:3的HCDR1氨基酸序列、SEQ ID NO:4的HCDR2氨基酸序列和SEQ IDNO:5的HCDR3氨基酸序列的重链可变区的互补决定区(CDR);和
(b)含有SEQ ID NO:6的LCDR1氨基酸序列、SEQ ID NO:7的LCDR2氨基酸序列和SEQ IDNO:8的LCDR3氨基酸序列的轻链可变区的CDR。
9.根据权利要求1所述的抗体或其抗原结合片段,所述抗体或其抗原结合片段包含:
(a)具有SEQ ID NO:13的HCDR1氨基酸序列、SEQ ID NO:14的HCDR2氨基酸序列和SEQID NO:15的HCDR3氨基酸序列的重链可变区的互补决定区(CDR);和
(b)含有SEQ ID NO:16的LCDR1氨基酸序列、SEQ ID NO:17的LCDR2氨基酸序列和SEQID NO:18的LCDR3氨基酸序列的轻链可变区的CDR。
10.根据权利要求1所述的抗体或其抗原结合片段,所述抗体或其抗原结合片段包含:
选自SEQ ID NO:9、19、43、47、51、55、59、63、67、71、75、79、83、87、91、95、99、103、107或111的重链可变区;和
选自SEQ ID NO:11、21、44、48、52、56、60、64、68、72、76、80、84、88、92、96、100、104、108或112的轻链可变区。
11.根据权利要求1所述的抗体或其抗原结合片段,所述抗体或其抗原结合片段包含由以保藏号KCLRF-BP-00417或KCLRF-BP-00418保藏的细胞系产生的抗体的互补决定区(CDR)。
12.一种药物组合物,其包含与药学上可接受的载体相关联的药学有效量的根据权利要求1所述的抗体或其抗原结合片段。
13.根据权利要求12所述的药物组合物,其进一步包含在化学疗法中使用的小分子抑制剂。
14.根据权利要求12所述的药物组合物,其进一步包含血管内皮生长因子(VEGF)拮抗剂。
15.根据权利要求14所述的药物组合物,其中所述VEGF拮抗剂是抗VEGF抗体、VEGF抑制性融合蛋白或小分子激酶抑制剂。
16.一种用于抑制患者中肿瘤生长的方法,其包括向所述患者给予包含根据权利要求12所述的抗体或抗原结合片段的药物组合物。
17.根据权利要求16所述的方法,其包括进一步给予在化学疗法中使用的小分子抑制剂或血管内皮生长因子(VEGF)拮抗剂。
18.根据权利要求17所述的方法,其中所述VEGF拮抗剂是抗VEGF抗体、VEGF抑制性融合蛋白或小分子激酶抑制剂。
19.一种用于在眼病患者中抑制脉络膜新血管形成、抑制眼血管渗漏或同时触发脉络膜毛细血管再生的方法,所述方法包括向所述患者给予根据权利要求12所述的药物组合物。
20.根据权利要求19所述的方法,其包括进一步给予在化学疗法中使用的小分子抑制剂或血管内皮生长因子(VEGF)拮抗剂。
21.根据权利要求20所述的方法,其中所述VEGF拮抗剂是抗VEGF抗体、VEGF抑制性融合蛋白或小分子激酶抑制剂。
22.根据权利要求19所述的方法,其中所述眼病是湿性年龄相关性黄斑变性(wAMD)、糖尿病性黄斑水肿(DME)或糖尿病性视网膜病(DR)。
23.一种核酸,其编码根据权利要求1所述的抗体或其抗原结合片段。
24.一种表达载体,其包含根据权利要求23所述的核酸。
25.一种用根据权利要求24所述的表达载体转化的宿主细胞。
26.一种用于产生抗Ang2抗体或其抗原片段的方法,所述方法包括培养根据权利要求25所述的宿主细胞。
Applications Claiming Priority (5)
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US201862633038P | 2018-02-20 | 2018-02-20 | |
US62/633,038 | 2018-02-20 | ||
KR1020190018769A KR102497171B1 (ko) | 2018-02-20 | 2019-02-18 | 항-안지오포이에틴-2(Ang2) 항체 및 그의 용도 |
KR10-2019-0018769 | 2019-02-18 | ||
PCT/KR2019/001983 WO2019164219A1 (en) | 2018-02-20 | 2019-02-19 | Anti-angiopoietin-2 antibodies and uses thereof |
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CN201980014282.1A Pending CN111741975A (zh) | 2018-02-20 | 2019-02-19 | 抗血管生成素-2抗体及其用途 |
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US (2) | US11498962B2 (zh) |
EP (1) | EP3755714A4 (zh) |
JP (2) | JP7366068B2 (zh) |
KR (1) | KR102497171B1 (zh) |
CN (1) | CN111741975A (zh) |
AU (1) | AU2019224694A1 (zh) |
CA (1) | CA3091613A1 (zh) |
MX (1) | MX2020008663A (zh) |
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KR20220002143A (ko) * | 2020-06-30 | 2022-01-06 | 삼성전자주식회사 | 항-Ang2 항체를 포함하는 코로나바이러스 감염증을 치료하기 위한 조성물 및 그의 용도 |
CN116209765A (zh) * | 2020-08-19 | 2023-06-02 | 药物抗体公司 | 经修饰抗体及其制备方法 |
JP2024517669A (ja) * | 2021-04-23 | 2024-04-23 | ニオテスバイオ インコーポレイテッド | アンジオポエチン-2に特異的に結合する抗体、またはその断片 |
KR102459211B1 (ko) * | 2021-04-23 | 2022-10-26 | (주)니오테스바이오 | 안지오포이에틴-2에 특이적으로 결합하는 항체, 또는 이의 단편 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101602806A (zh) * | 2003-11-26 | 2009-12-16 | 埃博马可西斯公司 | 针对血管内皮生长因子的人源化抗体 |
CN102257008A (zh) * | 2008-12-16 | 2011-11-23 | 霍夫曼-拉罗奇有限公司 | 针对人血管生成素2的抗体 |
US20130129722A1 (en) * | 2009-07-29 | 2013-05-23 | Regeneron Pharmaceuticals, Inc. | Methods for Treating Cancer by Administering an Anti-Ang-2 Antibody |
US20150030603A1 (en) * | 2013-07-29 | 2015-01-29 | Samsung Electronics Co., Ltd. | Anti-ang2 antibody |
Family Cites Families (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7658924B2 (en) | 2001-10-11 | 2010-02-09 | Amgen Inc. | Angiopoietin-2 specific binding agents |
RU2394839C2 (ru) | 2004-12-21 | 2010-07-20 | Астразенека Аб | Антитела против ангиопоэтина-2 и их применение |
JO3182B1 (ar) * | 2009-07-29 | 2018-03-08 | Regeneron Pharma | مضادات حيوية بشرية عالية الالفة مع تولد الاوعية البشرية - 2 |
CN103874709B (zh) | 2011-08-19 | 2016-12-21 | 瑞泽恩制药公司 | 抗tie2抗体及其用途 |
KR102146845B1 (ko) | 2013-07-30 | 2020-08-21 | 삼성전자주식회사 | 앤지오포이에틴-2에 특이적으로 결합하는 항체 및 그의 용도 |
KR102196450B1 (ko) | 2013-09-17 | 2020-12-30 | 삼성전자주식회사 | Tie2와 결합을 유도하는 항 Ang2 항체를 포함하는 항암제 |
US9994632B2 (en) | 2014-05-26 | 2018-06-12 | Samsung Electronics Co., Ltd. | Humanized or affinity-matured anti Ang-2 antibody and uses thereof |
CA2963606A1 (en) | 2014-11-10 | 2016-05-19 | F.Hoffmann-La Roche Ag | Anti-ang2 antibodies and methods of use |
-
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- 2019-02-18 KR KR1020190018769A patent/KR102497171B1/ko active IP Right Grant
- 2019-02-19 AU AU2019224694A patent/AU2019224694A1/en active Pending
- 2019-02-19 CA CA3091613A patent/CA3091613A1/en active Pending
- 2019-02-19 CN CN201980014282.1A patent/CN111741975A/zh active Pending
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- 2019-02-19 MX MX2020008663A patent/MX2020008663A/es unknown
- 2019-02-19 JP JP2020566520A patent/JP7366068B2/ja active Active
-
2020
- 2020-08-17 US US16/995,707 patent/US11498962B2/en active Active
-
2022
- 2022-10-12 US US18/045,964 patent/US20230272057A1/en active Pending
-
2023
- 2023-10-10 JP JP2023175308A patent/JP2024009914A/ja not_active Withdrawn
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101602806A (zh) * | 2003-11-26 | 2009-12-16 | 埃博马可西斯公司 | 针对血管内皮生长因子的人源化抗体 |
CN102257008A (zh) * | 2008-12-16 | 2011-11-23 | 霍夫曼-拉罗奇有限公司 | 针对人血管生成素2的抗体 |
US20130129722A1 (en) * | 2009-07-29 | 2013-05-23 | Regeneron Pharmaceuticals, Inc. | Methods for Treating Cancer by Administering an Anti-Ang-2 Antibody |
US20150030603A1 (en) * | 2013-07-29 | 2015-01-29 | Samsung Electronics Co., Ltd. | Anti-ang2 antibody |
Non-Patent Citations (1)
Title |
---|
JIN-SUNG PARK等: "Normalization of Tumor Vessels by Tie2 Activation and Ang2 Inhibition Enhances Drug Delivery and Produces a Favorable Tumor Microenvironment", CANCER CELL, vol. 30, no. 6, pages 953 - 967, XP029845233, DOI: 10.1016/j.ccell.2016.10.018 * |
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EP3755714A4 (en) | 2022-02-16 |
AU2019224694A1 (en) | 2020-08-06 |
US11498962B2 (en) | 2022-11-15 |
JP7366068B2 (ja) | 2023-10-20 |
KR102497171B1 (ko) | 2023-02-08 |
JP2024009914A (ja) | 2024-01-23 |
US20230272057A1 (en) | 2023-08-31 |
EP3755714A1 (en) | 2020-12-30 |
CA3091613A1 (en) | 2019-08-29 |
MX2020008663A (es) | 2021-02-09 |
US20210079083A1 (en) | 2021-03-18 |
JP2021514670A (ja) | 2021-06-17 |
KR20190100060A (ko) | 2019-08-28 |
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