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CN111093701A - 含有抗globo h抗体的抗体药物偶联物及其用途 - Google Patents

含有抗globo h抗体的抗体药物偶联物及其用途 Download PDF

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Publication number
CN111093701A
CN111093701A CN201880053150.5A CN201880053150A CN111093701A CN 111093701 A CN111093701 A CN 111093701A CN 201880053150 A CN201880053150 A CN 201880053150A CN 111093701 A CN111093701 A CN 111093701A
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China
Prior art keywords
antibody
globo
seq
immunoconjugate
cancer
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CN201880053150.5A
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Inventor
李照斌
庄士贤
徐铨龙
陈怡仁
聂瑜瑾
魏文胤
吴佳城
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Development Center for Biotechnology
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Development Center for Biotechnology
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
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Abstract

本发明提供包括抗Globo H抗体或其结合片段及治疗剂或标记的免疫偶联物，其具有式：Ab‑(L‑D)m，其中，Ab是该抗Globo H抗体或其结合片段，L是连接子或直接键，D是该治疗剂或该标记，且m是1至8的整数。该抗体可为人源化的单克隆抗体。本发明提供治疗癌症的方法，该方法包括向需要此治疗的个体给予医学上有效量的免疫偶联物，该免疫偶联物含有针对Globo H的抗体或其结合片段及与该抗体共价偶联的治疗剂。

Description

含有抗GLOBO H抗体的抗体药物偶联物及其用途
技术领域
本发明涉及含有Globo H抗体的抗体-药物偶联物及其治疗用途。
背景技术
抗体-药物偶联物(Antibody-drug conjugate，ADC)能够提供用于治疗诸如癌症的各种疾病或病况的靶向疗法。ADC是包含抗体的复合分子，该抗体连接至诸如细胞毒剂或药物的生物活性剂。借由组合抗体的独特靶向与药物的治疗作用，抗体-药物偶联物能够区分正常细胞与癌细胞，从而减少副作用。
ADC通常包含与抗体偶联的抗癌药物(例如，细胞毒素)，该抗体特异性靶向标志物(例如肿瘤标志物)。抗体在体内会追踪这些蛋白质且将自己结合于癌细胞表面。该抗体与该目标蛋白质(抗原)之间的结合会在肿瘤细胞中触发信号，之后肿瘤细胞将ADC内化。ADC内化之后，细胞毒性药物会被释放出来并杀死癌细胞。因为该药物具有特异性靶向，所以具有较低的副作用。
Globo H是一种六糖，属于许多与肿瘤相关的碳水化合物抗原，其可以在包括以下的各种上皮癌细胞表面上过度表达：乳腺癌细胞、结肠癌细胞、卵巢癌细胞、胰腺癌细胞、肺癌细胞及前列腺癌细胞。因此，Globo H是有前景的诊断/治疗靶点。
虽然针对Globo H的抗体是有用的，但仍然需要改进使用抗Globo H抗体的治疗剂。
发明内容
本发明是关于含有Globo H抗体的抗体-药物偶联物及其在疗法中的用途。
本发明一个方面涉及免疫偶联物。根据本发明一个实施方式的免疫偶联物包括抗Globo H抗体或其结合片段及治疗剂或标记，该偶联物具有式：Ab-(L-D)m，其中Ab是抗Globo H抗体或其结合片段，L是连接子或直接键，D是治疗剂或标记，且m是1至8的整数。
根据本发明的任何实施方式，Ab包含重链可变域和轻链可变域，重链可变域具有由HCDR1(GYISSDQILN，SEQ ID NO:1)、HCDR2(RIYPVTGVTQYXHKFVG，SEQ ID NO:2，其中X是任何氨基酸)及HCDR3(GETFDS，SEQ ID NO:3)组成的三个互补区，轻链可变域具有由LCDR1(KSNQNLLX'SGNRRYZLV，SEQ ID NO:4，其中X'是F、Y或W，且Z是C、G、S或T)、LCDR2(WASDRSF，SEQ ID NO:5)及LCDR3(QQHLDIPYT，SEQ ID NO:6)组成的三个互补区。
连接子L可为直接键，其中有效载荷(payload)D与抗体或其结合片段直接连接(共轭)。连接子可为蛋白质修饰或偶联中常用的任何连接子，诸如短肽(例如，gly-gly-gly)、短有机分子连接子(例如，SMCC，琥珀酰亚胺基-4(N-顺丁烯二酰亚胺基甲基)环己烷-1-甲酸酯)或类似连接子。
有效载荷D可为治疗剂，诸如细胞毒剂。可用于本发明的实施方式的细胞毒剂的实例可包括类美登素(maytansinoid)(例如，DM1或DM4)、单甲基奥瑞他汀E(monomethylauristatin E，MMAE)、单甲基奥瑞他汀F(monomethyl auristatin F，MMAF)、太平洋紫杉醇(paclitaxel)或类似物。
有效载荷D可为用于诊断或成像的标记或试剂。成像剂的实例可包括二伸乙三胺五乙酸(Diethylenetriaminepentaacetic acid，DTPA)或1,4,7,10-四氮杂环十二烷-1,4,7,10-四乙酸(1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid，DOTA)。
根据本发明一些实施方式，该抗体可为单克隆抗体，其可为人源化抗体。
本发明一个方面是诊断或成像可表达Globo H的细胞或组织的方法。根据本发明一个实施方式的方法包含向个体给予上述免疫偶联物。
本发明一个方面是治疗癌症的方法。根据本发明一个实施方式的方法包含向需要癌症治疗的个体给予医学上有效量的上述免疫偶联物。该癌症是上皮细胞癌，诸如乳腺癌、结肠癌、卵巢癌、胰腺癌、肺癌或前列腺癌。
本领域技术人员应了解，医学上有效量视诸多因素而定，诸如患者病症、年龄、疾病状态、给药途径等，且此有效量可基于常规实践的这些因素即可判定，无需过多的实验。
本发明的其他方式随以下说明将变得显而易见。
附图说明
图1显示根据本发明一个实施方式的ADC(DCB16001)的SDS-PGAE凝胶分析。结果显示，ADC在非还原条件及还原条件下保留适当抗体结构，亦即适当分子量。
图2显示HPLC曲线，指示Globo H抗体与SMCC-DM1之间的结合反应基本上是完全的且仅保留了残留量的Globo H抗体及SMCC-DM1。
图3说明本发明ADC(DCB16001)的MS分析的一个实施例，其指示各种数目的药物与抗体连接的分布，其中最丰富种类是具有1至8个药物与抗体连接。此样品中平均药物:抗体比(drug-to-antibody ratio，DAR)是4.07。
图4A及图4B显示ADC内化的荧光成像结果。结果指示，本发明的ADC可被表达GloboH的细胞(例如，MCF-7(图4B)及HCC-1428细胞(图4A))内化。
图5显示说明Meso Scale Discovery(MSD)电化学发光测定法的示意图，所述方法用于测量总的和偶联的抗Globo H抗体以进行DCBD16001的体内药代动力学研究。
图6显示DCBD16001的体内药代动力学研究结果。
图7显示在癌症异种移植模型中使用本发明ADC的治疗方案。
图8显示HCC-1428异种移植模型中的肿瘤生长抑制。
图9显示小鼠在图8的实验中的处理期间的体重变化。
具体实施方式
本发明的实施方式关于含有Globo H抗体的抗体-药物偶联物及其治疗用途。Globo H是一种六糖，属于许多与肿瘤相关的碳水化合物抗原，其可以在包括以下的各种上皮癌细胞表面上过度表达：乳腺癌细胞、结肠癌细胞、卵巢癌细胞、胰腺癌细胞、肺癌细胞及前列腺癌细胞。因此，以针对Globo H的抗体为基础的ADC是有用的诊断及/或治疗剂。
然而，治疗性抗体的快速内化或缺乏ADCC活性可能导致抗体无效以及产生抗性。因此，需要增强基于抗Globo H的治疗剂的治疗功效。一种方法是使有效载荷与抗Globo H抗体偶联(亦即，抗体-药物偶联物)。通过抗Globo H抗体与有效载荷偶联(亦即ADC)，本发明的实施方案比未偶联抗Globo H抗体更有效，从而能使用较少抗体。
根据本发明的实施方案，Globo H抗体或其结合片段可与药物、诊断剂或治疗剂偶联。因此，如本文所用的术语“抗体-药物偶联物”(ADC)可以指抗体部分(其可为全抗体或其结合片段)与有效载荷(其可为药物、诊断剂或治疗剂)偶联。
本发明ADC含有经设计用于治疗或诊断用途的有效载荷。相比于未偶联Globo H抗体，这些ADC具有较佳生物活性且仅需较小的量即可达成所期望的作用。
本发明的实施方式将借由以下特定实施例来说明。本领域技术人员应了解，这些实施例仅用于说明且其他修改及变化在不脱离本发明的范围的情况下是可能的。
实施例
除非另有指示，否则各1H NMR资料是在500MHz下获得。除非另外规定，否则本文中所用的缩写如下：
Bu：丁基；Bn：苯甲基；BOC：叔丁氧羰基；BOP：六氟磷酸苯并三唑-1-基氧基三/二甲胺基-鏻；DCC：二环己基碳化二亚胺；DMF：N,N-二甲基甲酰胺；DMAP：4-二甲胺基吡啶；EDC：1-(3-二甲胺基丙基)3-乙基碳化二亚胺盐酸盐；EtOAc：乙酸乙酯；Eq.：当量；HOBt：羟基苯并三唑；LAH：氢化锂铝；MeOH：甲醇；MHz：兆赫；MS(ES)：质谱仪-电子喷雾；NMP：N-甲基吡咯啶酮；Ph：苯基；Pr：丙基；TEA：三乙胺；THF：四氢呋喃；TLC：薄层层析；Tetrakis：四(三苯基膦)钯。
实施例1.制备抗Globo H抗体
根据本发明的实施方式，用于产生抗Globo H抗体的通用方法包括获得可产生针对Globo H的单克隆抗体的杂交瘤。用于产生单克隆抗体的方法为本领域中已知的且在此将不再赘述。简言之，小鼠是以抗原(Globo H)与适当的佐剂一起进行激发。随后，收集经免疫小鼠的脾细胞并与杂交瘤融合。使用任何已知方法(诸如ELISA)，通过结合Globo H抗原的能力而鉴别出阳性克隆。
本发明的抗体-药物偶联物(ADC)可特异性靶向Globo H。这些ADC可使用任何与Globo H特异性结合的抗体。举例而言，本发明ADC可以使用小鼠或人源化抗Globo H抗体或其scFv或Fab片段。示例性抗Globo H抗体可包含重链可变域和轻链可变域，所述重链可变域具有由HCDR1(GYISSDQILN，SEQ ID NO:1)、HCDR2(RIYPVTGVTQYXHKFVG，SEQ ID NO:2，其中X是任何氨基酸)及HCDR3(GETFDS，SEQ ID NO:3)组成的三个互补区，所述轻链可变域具有由LCDR1(KSNQNLLX'SGNRRYZLV，SEQ ID NO:4，其中X'是F、Y或W，且Z是C、G、S或T)、LCDR2(WASDRSF，SEQ ID NO:5)及LCDR3(QQHLDIPYT，SEQ ID NO:6)组成的三个互补区。
根据本发明的实施方式，抗体可为小鼠抗体。另一种选择是，抗体可为嵌合抗体(例如，人类恒定区与小鼠可变区偶联)或人源化抗体(例如，小鼠CDR移植于人类免疫球蛋白的框架区上)或完全人抗体。
单克隆抗体可通过以下方法进行人源化：从杂交瘤获得CDR序列并将CDR序列克隆至人类框架序列中以产生人源化抗体。可使用本领域中已知用于鉴别CDR序列的任何常见方法。本发明中的CDR区是用Kabat编号方案加以识别。首先，产生抗Globo H的杂交瘤(例如小鼠GBH杂交瘤)。此杂交瘤可借由用于产生单克隆抗体的标准方案产生。随后例如使用
Figure BDA0002384209660000051
试剂分离出杂交瘤的总RNA。随后例如使用第一链cDNA合成试剂盒(SuperscriptIII)及寡聚(dT20)引物或Ig-3'恒定区引物，从总RNA合成cDNA。
随后从cDNA克隆免疫球蛋白基因的重链及轻链可变区。举例而言，借由PCR，使用小鼠Ig-5'引物组(Novagen)，从小鼠GBH杂交瘤cDNA扩增抗Globo H mAb的VH及VL可变区。该多个PCR产物可使用CloneJetTM PCR克隆试剂盒(Ferments)直接克隆至适合载体(例如pJET1.2载体)中。pJET1.2载体含有致死性插入序列段且仅在所需基因克隆至此致死性区域中时才会在选择条件下存活。这有助于重组菌落的选择。最后，针对所需克隆筛选重组菌落，分离那些克隆的DNA并测序。可在国际ImMunoGeneTics信息系统(IGMT)网站分析免疫球蛋白(IG)核苷酸序列。
抗体表达及纯化
为产生抗体，该多个经分离的克隆可在任何适合的细胞中表达。举例而言，F293细胞(Life technologies)经表达抗Globo H mAb的质粒转染且培养7天。使用蛋白A亲和柱(GE)，从培养基纯化出抗Globo H抗体。使用本领域中已知的程序或根据制造商的说明，以Bio-Rad蛋白质分析试剂盒测定蛋白质浓度及用12％SDS-PAGE进行分析。
根据本发明的实施方式，可使用这些抗Globo H抗体中的任一制备抗体-药物偶联物(ADC)，如以下实施例中所说明。
实施例2.制备抗Globo H抗体-生物素偶联物
Figure BDA0002384209660000061
作为药物与抗体偶联的实施例，可使用生物素-抗生物素蛋白(avidin)系统来研究反应条件并说明ADC策略的可操作性。在此特定实施例中，使用生物素类似物(亦即，生物素-N-琥珀酰亚胺酯)作为偶联剂与抗体上的氨基反应。该氨基可为抗体上的赖氨酸残基的侧链。
简言之，向抗Globo H单克隆抗体430μL(2.7mg/ml)于缓冲液(50mM磷酸钾、50mM氯化钠、2mM EDTA；pH 6.5)中的溶液中缓慢添加3.8μL Osu-生物素(生物素-O-琥珀酰亚胺；20mM，于DMSO中)。在氩气下在室温下搅拌反应混合物且分别搅拌2、4及16小时。使用配置有标称分子量限值(NMWL)为30kDa的Amicon Ultra-15离心过滤装置在pH 7.4PBS缓冲液中将抗体制剂进行脱盐及浓缩，得到Globo H-生物素ADC 2-1。
Figure BDA0002384209660000071
在替代方法中，该生物素类似物是与抗体上的半胱氨酸残基的SH基偶联。如以上反应流程中所示，向抗Globo H单克隆抗体180μL(5.0mg/mL，于pH 7.4PBS缓冲液中)的溶液中缓慢添加TCEP(5.0当量)且在37℃下搅拌1.0小时。随后向反应混合物中添加生物素-顺丁烯二酰亚胺(12当量)且在氩气下在室温下搅拌20小时。使用配置有NMWL为30kDa的Amicon Ultra-15离心过滤装置在pH 7.4PBS缓冲液中将抗体制剂进行脱盐及浓缩，得到Globo H-生物素ADC 2-2。
实施例3.制备抗Globo H抗体-SMCC-DM1偶联物
Figure BDA0002384209660000072
在此实施例中，ADC含有DM1，DM1是为了癌症疗法而开发的类美登素。美登素(Maytansine)，一种苯并桥环巨环内酯(benzoansamacrolide)，是非常有效的靶向微管的化合物，其可诱导有丝分裂阻滞并在亚纳摩尔(subnanomolar)浓度下杀死肿瘤细胞。DM1在微管的尖端结合以抑制微管的动态性，亦即，抑制微管的组装。DM1是一种全身性毒性比美登素低的类美登素。在此实施例例中，SMCC-DM1是含有反应性连接子SMCC的DM1，其用于与抗体反应以制造抗体药物偶联物。SMCC-DM1可自商业来源获得，诸如MedKoo Biosciences公司或ALB Technology。
举例而言，向抗Globo H单克隆抗体500μL(2.9mg/mL)于缓冲液(50mM磷酸钾、50mM氯化钠、2mM EDTA；pH 6.5)中的溶液中缓慢添加58μL SMCC-DM1(5mM，于DMSO中)。在氩气下在37℃下搅拌反应混合物且搅拌20小时。使用配置有NMWL为30kDa的Amicon Ultra-15离心过滤装置在pH 7.4PBS缓冲液中将抗体制剂进行脱盐及浓缩，得到抗Globo H-SMCC-DM1ADC 3。(DCBD16001)
实施例4.制备抗Globo H抗体-SMCC-DM4偶联物
Figure BDA0002384209660000081
DM4是另一种美登素类似物。DM4也为抑制处于有丝分裂的细胞增殖的有效靶向微管的化合物。本发明的一些实施方式可以使用DM4。在此实施例中，向抗Globo H单克隆抗体500μL(2.9mg/mL)于缓冲液(50mM磷酸钾、50mM氯化钠、2mM EDTA；pH 6.5)中的溶液中缓慢添加58μL SMCC-DM4(5mM，于DMSO中)。在氩气下在37℃下搅拌反应混合物且搅拌20小时。使用配置有NMWL为30kDa的Amicon Ultra-15离心过滤装置在pH 7.4PBS缓冲液中将抗体制剂进行脱盐及浓缩，得到抗Globo H-SMCC-DM4 ADC 4。
实施例5.制备抗Globo H抗体-MMAE偶联物
Figure BDA0002384209660000082
单甲基奥瑞他汀E(MMAE)是一种抗恶性肿瘤药(antineoplastic agent)；其借由阻断微管蛋白的聚合来抑制细胞分裂。其衍生自出现于海洋无壳软件动物中的肽(海兔毒素(dolastatin))。已证明MMAE是ADC的有用有效载荷。
在此实施例中，向抗Globo H单克隆抗体400μL(5.0mg/mL)于缓冲液(50mM磷酸钾、50mM氯化钠、2mM EDTA；pH 6.5)中的溶液中缓慢添加40μL OSu-MMAE(5mM，于DMSO中)。在氩气下在37℃下搅拌反应混合物且搅拌20小时。使用配置有NMWL为30kDa的Amicon Ultra-15离心过滤装置在pH 7.4PBS缓冲液中将抗体制剂进行脱盐及浓缩，得到抗Globo H-MMAEADC 5。
实施例6.制备抗Globo H抗体-vc-MMAE偶联物
Figure BDA0002384209660000091
ADC中的连接子会对生物活性具有显著影响。举例而言，体内研究证实，经肽连接的偶联物会诱导经建立的肿瘤异种移植物的消退及治愈，其中治疗指数高达60倍。这些偶联物说明了连接子技术、药物效力及偶联方法在开发用于癌症疗法的安全及有效的mAb-药物偶联物中的重要性。
本发明的一些实施方式是关于经由溶酶体可裂解二肽缬氨酸-瓜氨酸(valine-citrulline，vc)连接于抗体的MMAE，已证明，该二肽可改良ADC功效。在此实施例中，向抗Globo H单克隆抗体400μL(5.0mg/mL)于缓冲液(50mM磷酸钾、50mM氯化钠、2mM EDTA；pH6.5)中的溶液中缓慢添加40μL OSu-vc-MMAE(5mM，于DMSO中)。在氩气下在37℃下搅拌反应混合物且搅拌20小时。使用配置有NMWL为30kDa的Amicon Ultra-15离心过滤装置在pH7.4PBS缓冲液中将抗体制剂进行脱盐及浓缩，得到抗Globo H-vc-MMAE ADC 6。
实施例7.制备抗Globo H抗体-MMAF偶联物
Figure BDA0002384209660000092
本发明的一些实施方式是关于含有单甲基奥瑞他汀F(MMAF)的ADC，MMAF是MMAE的类似物。向抗Globo H单克隆抗体400μL(5.0mg/mL)于缓冲液(50mM磷酸钾、50mM氯化钠、2mMEDTA；pH 6.5)中的溶液中缓慢添加40μL OSu-MMAF(5mM，于DMSO中)。在氩气下在37℃下搅拌反应混合物且搅拌20小时。使用配置有NMWL为30kDa的Amicon Ultra-15离心过滤装置在pH 7..4PBS缓冲液中将抗体制剂进行脱盐及浓缩，得到抗Globo H-MMAF ADC 7。
实例8.制备抗Globo H抗体-4-异硫氰基-苯基-DTPA偶联物
Figure BDA0002384209660000101
ADC除了治疗之外，也可用于诊断及/或成像。本发明的一些实施方式涉及能够与目标分子特异性结合的成像试剂。举例而言，为了成像，ADC的有效载荷可含有用于结合所选择的金属(诸如放射性过渡金属)的螯合官能团。本领域中已知多种用于诊断及/或成像用途的螯合官能团，诸如二伸乙三胺五乙酸(DTPA)或1,4,7,10-四氮杂环十二烷-1,4,7,10-四乙酸(DOTA)。DTPA是具有二伸乙基三胺主链及5个羧基的氨基聚羧酸。DTPA可视为EDTA的延伸版本。此处将使用DTPA作为实施例。本领域技术人员应了解，在不脱离本发明的范围的情况下也可使用其他螯合剂。
在此实施例中，向抗Globo H单克隆抗体400μL(5.0mg/mL)于缓冲液(10mM磷酸钠；pH 8.0)中的溶液中缓慢添加40μL 4-异硫氰基-苯基-DTPA(5mM，于DMSO中)。在氩气下在37℃下搅拌反应混合物且搅拌20小时。使用配置有NMWL为30kDa的Amicon Ultra-15离心过滤装置在pH 7.4PBS缓冲液中将抗体制剂进行脱盐及浓缩，得到抗Globo H-硫脲-4-苯基-DTPA ADC 8。
实施例9.制备抗Globo H抗体-N-苯基己二酰二胺-DTPA偶联物
Figure BDA0002384209660000111
在此实施例中，DPTA经由连接子己二酰二胺连接于抗体。向抗Globo H单克隆抗体400μL(5.0mg/mL)于缓冲液(10mM磷酸钠；pH 8.0)中的溶液中缓慢添加40μL 6-OSu-N-苯基己酰胺-DPTA(5mM，于DMSO中)。在氩气下在37℃下搅拌反应混合物且搅拌20小时。使用配置有NMWL为30kDa的Amicon Ultra-15离心过滤装置在pH 7.4PBS缓冲液中将抗体制剂进行脱盐及浓缩，得到抗Globo H-N-苯基己酰胺-DPTA ADC 9。
实施例10.SDS-PAGE
本发明的各种ADC可借由本领域中已知的技术进行分析，诸如SDS-PAGE及HPLC。举例而言，从实施例1和3获得的抗Globo H mAb及抗Globo H ADC 3(DCBD16001)的溶液借由使用4-12％非还原性及还原性SDS-PAGE凝胶、继之以考马斯亮蓝(Coomassie brilliantblue)染色来分析。图1显示，ADC(DCB16001)在非还原条件及还原条件下保留适当抗体结构，即适当分子量。
实施例11.PLRP-HPLC
本发明的各种ADC可通过HPLC来分析。图2显示，偶联反应基本上是完全的且仅剩下残留量的Globo H抗体及SMCC-DM1。
实施例12.有效载荷耦合分析
评估药物:抗体比(DAR)对于监测目标抗体上的有效载荷偶联效率是重要的。药物:抗体比会影响抗Globo H ADC产物的治疗功效。液相层析-质谱法(Liquidchromatography-mass spectrometry，LC-MS)是用于测定赖氨酸连接的抗体-药物偶联物(ADC)的药物:抗体比(DAR)及载药量分布(drug load distribution)的优良方法。峰面积百分比表示特定载药ADC种类的相对分布。随后使用峰面积百分比信息及载药数目计算加权平均DAR。
图3说明本发明ADC(抗Globo H-SMCC-DM1 ADC 3.(DCBD16001))的MS分析的一个实施例，其指示各种数目的药物与抗体连接的分布，其中最丰富种类是具有1至8个药物与抗体连接。此样品中的平均药物:抗体比(DAR)是4.07。多个药物复本与一个抗体连接将确保药物更有效地递送至细胞中。
实施例13.结合亲和力
本发明ADC的结合亲和力可借由本领域中已知的任何适合方法评定，诸如BIAcore或ELISA。在此实施例中，使用BIAcore来测量本发明ADC的亲和力。
简言之，制备抗Globo H ADC的流动溶液以进行结合动力学研究。将配体Globo H固定在CM5芯片上：首先，在固定缓冲液(10mM乙酸钠，pH 4.5)中，将配体(Globo H-胺)稀释至6mg/mL。一般在25℃下使用5μL/min的流速固定。胺偶联试剂盒中提供了用于固定的试剂。活化：EDC/NHS 7分钟。固定：流动时间720秒。失活：1.0M乙醇胺，pH 8.5，7分钟。此过程将在传感器芯片CM5上产生约200RU的反应结合水平。
随后，如下进行单循环动力学分析：Biacore单循环动力学(single-cyclekinetics，SCK)方法具有用以获得动力学数据的软件。选择操作(Run)：方法。参数设定如下：数据收集速率：1Hz；检测模式：双重；温度：25℃；浓度单位：nM；缓冲液A：HBS-EP+缓冲液。选择启动(Start up)且将复制数改为3。选择启动循环(Startup cycle)且参数设定如下：类型：低样品消耗；接触时间：150秒；解离时间：420秒；流速：50μL/min；流动路径：两个。选择样品循环(Sample cycle)且参数设定如下：类型：单循环动力学；浓度/循环：5；接触时间：150秒；解离时间：420秒；流速：50μl/min；流动路径：两个。选择再生(Regeneration)且参数设定如下：再生溶液：10mM甘氨酸pH2.0/1.5(v/v＝1)；接触时间：45秒；流速：30μL/min；流动路径：两个。选择样品的复本(Copy)且参数设定如上。制备样品：将分析物抗体DCBPR1101在操作缓冲液中稀释至200nM。从200nM样品制备浓度系列：将200μL的200nM溶液与200μL操作缓冲液混合，得到100nM溶液。继续连续稀释以获得以下：200、100、50、25及12.5nM。制备样品且根据支架位置(Rack Position)定位。根据“准备运行协议”(PrepareRun Protocol)，确保一切正确，然后单击“开始”(Start)以开始实验。使用由Biacore T100评估软件2.0提供的预定模型(1:1结合)进行亲和力结合曲线拟合。
表1汇总了本发明的一个ADC(DCBD16001)的BIAcore分析结果。这些结果显示，本发明ADC仍然能够与抗原(Globo H)特异性结合。
表1
k<sub>on</sub>(1/Ms) k<sub>off</sub>(1/s) K<sub>D</sub>(M) R<sub>max</sub> Chi<sup>2</sup>
8.725E+4 1.427E-3 1.636E-8 365.6 7.04
实施例14.细胞毒性测定
含有细胞毒性有效载荷的本发明ADC可用于杀死目标细胞，诸如杀死癌细胞。为此分析，从ATCC获得MCF-7、HCC-1428、BT-474。所有细胞系在5％CO2的湿润孵育箱氛围中于37℃下在适合培养基中培养。所有细胞系传代培养至少三代，将细胞种植于96孔黑色平底板(所有细胞系均为10,000个细胞/100微升/孔)中，且使细胞在5％CO2的潮湿气氛中于37℃下进行细胞粘附过夜。
ADC是从1μg/μl储备溶液制备且在细胞接种之后24小时稀释至适当的操作浓度。首先用PBS将20mM测试物品稀释成最高操作浓缩物。用DMSO对此最高操作浓缩物进行八个点的连续三倍稀释，且随后另外用培养基稀释100×。检测8种浓缩物，最终浓度在100nM至0.046nM(DM-1)或66.67nM至0.030nM(DBCD16001、DCBPR1101)的范围内。最终PBS浓度是1％。随后移除培养基且替换成含有不同浓度的游离DM1、DBCD16001、DCBPR1101及200微升/孔的新鲜培养基且孵育细胞72小时。在室温下预解冻Cell Titer-Glo试剂48小时。在此时间点之后，移除培养基且在室温下向孔中添加100微升/孔的Cell Titer-Glo试剂(PromegaG7571，批次0000182872)，持续10分钟，且使用CLARIO
Figure BDA0002384209660000131
高效单色仪多模式酶标仪来测量发光信号。关于所有细胞分析，使用GraphPad Prism 6三参数曲线拟合来产生剂量反应曲线。
这些分析的结果显示于下表2中。显然，相比于抗原阴性细胞(即BT-474)，本发明ADC(亦即DCBD16001)可有效杀死抗原(Globo H)表达细胞(MCF-7及HCC-1428)。
表2
Figure BDA0002384209660000141
实施例15.内化分析
抗体经受体介导的内化作用可提供细胞特异性药物递送。内化是某些使用ADC的靶向疗法所需要的。本领域中已知，ADC内化是抗体依赖性及有效载荷依赖性的。亦即，并非所有抗体都能为ADC提供递送机制。类似地，不同有效载荷在相同抗体上会具有显著不同的内化效率。
抗Globo H ADC及抗Globo H抗体的内化及降解可借由流式细胞仪测量。在这项研究中可使用两种方法，直接在抗体上进行荧光标记或使用经荧光偶联的第二抗体来检测内化后留在细胞表面上的第一抗体。
简言之，以1×105个细胞/孔接种MCF7或HCC 1428乳腺癌细胞。接着，使0.5-1mg经荧光标记的抗Globo H抗体或抗Globo H ADC在FACS缓冲液中在4℃下经受100μl目标细胞(细胞密度1×106/ml)作用1小时，以实现抗Globo H抗体或抗Globo H ADC与细胞表面目标的特异性结合。在孵育之后，用FACS缓冲液洗涤细胞三次以移除未结合抗体。随后在37℃下在5％CO2下孵育细胞以进行抗体内化。
在不同时间点，用胰蛋白酶解离细胞且用经荧光偶联抗人类IgG第二抗体染色5分钟，随后使用Beckman流式细胞仪系统进行分析。关于经间接标记的抗体或经间接荧光标记的第二抗体的内化及降解的测量，如相比于4℃对照组(作为抗体内化背景)，通过Beckman流式细胞仪软件分析在各时间点的荧光强度及细胞结合百分比。各实验重复三次。
为了通过荧光成像研究抗Globo H ADC及抗Globo H抗体的内化，使用标准滤光片配置将细胞在
Figure BDA0002384209660000151
Core显微镜上成像。简言之，MCF-7或HCC1428目标细胞以约2×105个细胞/孔在8孔玻璃盖玻片底皿(Nunc)上传代培养。在附着之后，细胞在37℃下与10nM抗Globo H抗体或与Alexa-488偶联的抗Globo H ADC一起孵育过夜。在内化之前，用Rab5-mCherry或LAMP1-mApple质粒转染细胞以便用荧光标记物LysoTracker进行核内体及溶酶体标记。对细胞进行洗涤且在Deltavision反褶积显微镜(deconvolutionmicroscope)上成像以确定Alexa 488和Alexa594共定位。
图4A及图4B显示这些研究的结果。结果指示，本发明的ADC可被表达Globo H的细胞(例如，MCF-7(图4B)及HCC-1428细胞(图4A))内化。
实施例16.体内PK
此研究使用Meso Scale Discovery(MSD)电化学发光(Electrochemiluminescent，ECL)法对BALB/c小鼠样品中的DCBD 16001进行药代动力学分析。抗体MSD测定法可测量偶联和未偶联的抗体，如图5所示。如此实施例所示，或在总抗体测定中，板涂有山羊抗人类IgG，可捕捉所有人源化抗体(偶联和未偶联的)。关于偶联抗体分析，板涂有针对有效载荷(药物)的抗体，诸如图5中所示的抗美登素抗体。
以1mg/kg的剂量经由尾部静脉对BALB/c小鼠给药。随后在不同时间点获得血液样品以借由MESO QuickPlex SQ 120方法测定BALB/c小鼠中的DCBD16001的浓度。DCBD16001的药代动力学参数借由非房室分析(noncompartmental analysis)，使用6.3版PhoenixTMWinNonlin程序来分析。
图6显示体内药代动力学研究的结果且下表3汇总了PK研究的结果。总量抗体MSD分析：测量偶联及未偶联抗体两者。偶联抗体MSD分析：仅测量偶联抗体。DCBD16001的体内半衰期是约127小时，其相比于已知治疗性抗体的体内半衰期而言相对较短，表明若出现严重副作用，则DCBD16001可被较快排出。
表3
Figure BDA0002384209660000161
实施例17.抗Globo H ADC的异种移植模型
本发明的一些实施例是关于使用ADC中的抗体作为归巢/靶向剂进行诊断、成像及治疗疾病的方法。该抗体部分将与其目标抗原特异性结合，而有效载荷将提供诊断/成像或治疗试剂。可能的诊断/成像试剂可包括例如荧光部分或放射性探针，而治疗试剂可包括例如细胞毒剂或免疫调节剂(例如，CD3)。
在此特定实施例中，评定本发明ADC治疗癌症的能力。简言之，从BioLasco TaiwanCo.,LTD.购得六至七周龄雄性CB.17SCID小鼠，且隔离检疫一周。在实验期内，将5只小鼠圈养在一个笼中。所有动物均以12-小时光/12-小时暗循环托管于Da-Hu动物研究室中19-25℃下。动物可以自由地随意取食啮齿动物颗粒状食物及水。动物研究的实验方案是经实验动物护理和使用委员会(Institutional Animal Care and Use Committee)、DCB审查及批准。
在注射肿瘤细胞前7天，给小鼠接种17-β-雌二醇沉淀(0.18mg/粒，60天释放；美国创新研究所(Innovative Research of America,USA))。在对数期生长期间收集用于植入的HCC1428乳腺癌细胞且将其再悬浮于磷酸盐缓冲盐水(phosphate buffered saline，PBS)中。在各小鼠的侧腹皮下(s.c.)注射0.15mL 50％基质胶溶液(Matrigel solution)(BD Biosciences,MA,USA)中的1×107个HCC1428细胞。当平均肿瘤体积达到250mm3时，将小鼠随机分成8组。第1组接受PBS，充当对照组，用于计算肿瘤生长抑制率。第2组至第5组分别按以下接受DCBD16001：3mg/kg(mpk)每周两次、10mpk每周两次、20mpk每周一次及在第1天及第11天各30mg/kg。第6组接受未偶联抗体PR1101(无有效载荷)，10mpk每周两次。第7组中的小鼠接受同型抗体，10mpk每周两次，充当阴性对照组。第8组中的小鼠接受太平洋紫杉醇，10mpk每周一次，充当阳性治疗对照组。经载剂、DCBD16001、未偶联抗体及同型抗体处理的小鼠经由静脉内注射，而太平洋紫杉醇通过腹膜内注射给予。
肿瘤体积使用测径规每周测量三次且使用下式估算：肿瘤体积＝(w2×l)/2，其中w＝肿瘤的宽度且l＝直径长度(mm)。肿瘤生长抑制(tumor growth inhibition，TGI)的百分比使用下式计算：TGI％＝[1-(T/C)]×100％，其中T及C分别表示处理组及对照组的平均肿瘤体积。
给动物称重，每周三次，直至研究完成。体重变化按相比于初始体重的体重增加百分比计算。
图7说明实验方案及治疗方案。在此实验中，使用HCC1428细胞，其是乳腺癌细胞系。这些细胞会在其表面上表达Glob H。
图8显示体内异种移植肿瘤治疗研究的结果。如所示，10mg/kg、20mg/kg及30mg/kg的本发明ADC(DCBD16001)明显比未偶联抗体(抗Globo H抗体)更有效。结果指示本发明ADC的有效性。明显改良的功效是因为该经偶联的有效载荷，其可以增强细胞毒性和/或改良药代动力学。
引人注意的是，在第1天及第11天用30mpg DCBD16001处理(总剂量60mg/kg)是最有效的，明显比10mg/kg每周两次(总剂量60mg/kg)更有效。此结果指示，在较高的单一注射剂量及较长的给药时间间隔(10天)下，ADC更有效。在相同的总剂量下，较长的给药时间间隔(例如，10天)产生较佳的结果，这一事实出人意料。这表明，对于抗Globo H ADC而言，较高的Cmax将比较高的AUC更重要。
以下事实证实肿瘤生长抑制是由增强的肿瘤细胞杀死或肿瘤细胞生长抑制引起：不同处理组中的小鼠的体重没有明显变化，如图9中所示。
以上实施例清楚地说明了获得并表征本发明ADC的各种方法，以及本发明ADC在治疗癌症方面的有效性。即使本发明的实施例用有限数目的实例加以说明，本领域技术人员仍应了解，其他变化及修改在不脱离本发明的范围的情况下是可能的。因此，本发明的范围应仅受所附权利要求书限制。
序列表
<110> 财团法人生物技术开发中心
DCB-USA LLC
<120> 含有抗GLOBO H抗体的抗体药物偶联物及其用途
<130> DCB013-724PCT
<150> US 62/520,484
<151> 2017-06-15
<160> 6
<170> PatentIn version 3.5
<210> 1
<211> 10
<212> PRT
<213> 小鼠(Mus musculus)
<400> 1
Gly Tyr Ile Ser Ser Asp Gln Ile Leu Asn
1 5 10
<210> 2
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 合成的
<220>
<221> MISC_FEATURE
<222> (12)..(12)
<223> X是任何氨基酸
<400> 2
Arg Ile Tyr Pro Val Thr Gly Val Thr Gln Tyr Xaa His Lys Phe Val
1 5 10 15
Gly
<210> 3
<211> 6
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 合成的
<400> 3
Gly Glu Thr Phe Asp Ser
1 5
<210> 4
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 合成的
<220>
<221> MISC_FEATURE
<222> (8)..(8)
<223> 8位的X是F, Y, 或W
<220>
<221> MISC_FEATURE
<222> (15)..(15)
<223> 15位的X是C, G, S, 或T
<400> 4
Lys Ser Asn Gln Asn Leu Leu Xaa Ser Gly Asn Arg Arg Tyr Xaa Leu
1 5 10 15
Val
<210> 5
<211> 7
<212> PRT
<213> 小鼠(Mus musculus)
<400> 5
Trp Ala Ser Asp Arg Ser Phe
1 5
<210> 6
<211> 9
<212> PRT
<213> 小鼠(Mus musculus)
<400> 6
Gln Gln His Leu Asp Ile Pro Tyr Thr
1 5

Claims (12)

1.一种与Globo H特异性结合的免疫偶联物，包含：
抗Globo H抗体或其结合片段，及
治疗剂或标记，
其中所述免疫偶联物具有式：Ab-(L-D)m，
其中Ab是所述抗Globo H抗体或其结合片段，L是连接子或直接键，D是所述治疗剂或所述标记，且m是1至8的整数。
2.如权利要求1所述的免疫偶联物，其特征在于，所述抗体是单克隆抗体。
3.如权利要求1所述的免疫偶联物，其特征在于，所述抗体是人源化抗体。
4.如权利要求1所述的免疫偶联物，其特征在于，所述治疗剂是细胞毒剂。
5.如权利要求4所述的免疫偶联物，其特征在于，所述细胞毒剂是类美登素1(DM1)、类美登素4(DM4)、单甲基奥瑞他汀E(MMAE)、单甲基奥瑞他汀F(MMAF)或太平洋紫杉醇。
6.如权利要求1所述的免疫偶联物，其特征在于，D是诊断或成像试剂。
7.如权利要求1至6中任一项所述的免疫偶联物，其特征在于，所述抗体包含重链可变域和轻链可变域，所述重链可变域具有由HCDR1(GYISSDQILN，SEQ ID NO:1)、HCDR2(RIYPVTGVTQYXHKFVG，SEQ ID NO:2，其中X是任何氨基酸)及HCDR3(GETFDS，SEQ ID NO:3)组成的三个互补区，所述轻链可变域具有由LCDR1(KSNQNLLX'SGNRRYZLV，SEQ ID NO:4，其中X'是F、Y或W，且Z是C、G、S或T)、LCDR2(WASDRSF，SEQ ID NO:5)及LCDR3(QQHLDIPYT，SEQID NO:6)组成的三个互补区。
8.一种用于诊断或成像表达Globo H的细胞或组织的组合物，包含权利要求6所述的免疫偶联物。
9.一种用于治疗癌症的药物组合物，包含权利要求1至5中任一项所述的免疫偶联物。
10.如权利要求9所述的药物组合物，其特征在于，所述抗体包含重链可变域和轻链可变域，所述重链可变域具有由HCDR1(GYISSDQILN，SEQ ID NO:1)、HCDR2(RIYPVTGVTQYXHKFVG，SEQ ID NO:2，其中X是任何氨基酸)及HCDR3(GETFDS，SEQ ID NO:3)组成的三个互补区，所述轻链可变域具有由LCDR1(KSNQNLLX'SGNRRYZLV，SEQ ID NO:4，其中X'是F、Y或W，且Z是C、G、S或T)、LCDR2(WASDRSF，SEQ ID NO:5)及LCDR3(QQHLDIPYT，SEQID NO:6)组成的三个互补区。
11.如权利要求9所述的药物组合物，其特征在于，所述癌症是上皮细胞癌。
12.如权利要求11所述的药物组合物，其特征在于，所述癌症是乳腺癌、结肠癌、卵巢癌、胰腺癌、肺癌或前列腺癌。
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021228044A1 (en) * 2020-05-11 2021-11-18 Hunilife Biotechnology, Inc. Drug conjugates containing alpha-enolase antibodies and uses thereof
WO2023020624A1 (en) * 2021-08-20 2023-02-23 Hunilife Biotechnology, Inc. Use of alpha-enolase antagonist in treating fibrotic diseases
WO2023025243A1 (zh) * 2021-08-27 2023-03-02 石药集团巨石生物制药有限公司 抗Nectin-4抗体、药物缀合物及其制备方法和用途

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TWI782930B (zh) * 2016-11-16 2022-11-11 美商再生元醫藥公司 抗ｍｅｔ抗體，結合ｍｅｔ之雙特異性抗原結合分子及其使用方法
CN110290800A (zh) * 2016-11-21 2019-09-27 台湾浩鼎生技股份有限公司 缀合生物分子、医药组合物及方法
WO2022206806A1 (zh) * 2021-03-31 2022-10-06 荣昌生物制药(烟台)股份有限公司 一种采用lc-ms/ms检测adc中dtpa含量的方法

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20160102151A1 (en) * 2014-01-16 2016-04-14 Academia Sinica Compositions and methods for treatment and detection of cancers
CN105764526A (zh) * 2013-08-26 2016-07-13 马布瓦克斯医疗公司 编码抗唾液酸化路易斯a抗原的人抗体的核酸
US20160274121A1 (en) * 2015-01-16 2016-09-22 Academia Sinica Cancer markers and methods of use thereof
US20160289340A1 (en) * 2015-01-16 2016-10-06 Academia Sinica Compositions and methods for treatment and detection of cancers
CN106188293A (zh) * 2015-04-17 2016-12-07 江苏恒瑞医药股份有限公司 抗c-Met抗体和抗c-Met抗体-细胞毒性药物偶联物及其医药用途
CN106456766A (zh) * 2014-03-19 2017-02-22 台湾基督长老教会马偕医疗财团法人马偕纪念医院 抗免疫原性糖肽的抗体、包含其的组合物及其用途

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990005144A1 (en) * 1988-11-11 1990-05-17 Medical Research Council Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors
CA2490399C (en) * 2002-06-19 2015-10-06 Raven Biotechnologies, Inc. Novel raag10 cell surface target and a family of antibodies recognizing that target
EP2897980B1 (en) * 2012-09-19 2019-11-06 Innate Pharma Kir3dl2 binding agents
WO2014087007A1 (en) * 2012-12-06 2014-06-12 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Plasmodium falciparum neutralizing antibodies
WO2015014376A1 (en) 2013-07-31 2015-02-05 Biontech Ag Diagnosis and therapy of cancer involving cancer stem cells
WO2016114819A1 (en) 2015-01-16 2016-07-21 Academia Sinica Compositions and methods for treatment and detection of cancers
JP2017507118A (ja) * 2014-01-16 2017-03-16 アカデミア シニカＡｃａｄｅｍｉａ Ｓｉｎｉｃａ がんの処置および検出のための組成物および方法
TW201825522A (zh) * 2016-09-23 2018-07-16 張志隆 抗-globo h 抗體
CA3064443A1 (en) * 2017-05-24 2018-11-29 Development Center For Biotechnology Humanized antibodies against globo h and uses thereof in cancer treatments

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105764526A (zh) * 2013-08-26 2016-07-13 马布瓦克斯医疗公司 编码抗唾液酸化路易斯a抗原的人抗体的核酸
US20160102151A1 (en) * 2014-01-16 2016-04-14 Academia Sinica Compositions and methods for treatment and detection of cancers
CN106456766A (zh) * 2014-03-19 2017-02-22 台湾基督长老教会马偕医疗财团法人马偕纪念医院 抗免疫原性糖肽的抗体、包含其的组合物及其用途
US20160274121A1 (en) * 2015-01-16 2016-09-22 Academia Sinica Cancer markers and methods of use thereof
US20160289340A1 (en) * 2015-01-16 2016-10-06 Academia Sinica Compositions and methods for treatment and detection of cancers
CN106188293A (zh) * 2015-04-17 2016-12-07 江苏恒瑞医药股份有限公司 抗c-Met抗体和抗c-Met抗体-细胞毒性药物偶联物及其医药用途

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021228044A1 (en) * 2020-05-11 2021-11-18 Hunilife Biotechnology, Inc. Drug conjugates containing alpha-enolase antibodies and uses thereof
WO2023020624A1 (en) * 2021-08-20 2023-02-23 Hunilife Biotechnology, Inc. Use of alpha-enolase antagonist in treating fibrotic diseases
TWI834254B (zh) * 2021-08-20 2024-03-01 上毅生物科技股份有限公司 α-烯醇酶拮抗劑治療纖維化疾病之用途
WO2023025243A1 (zh) * 2021-08-27 2023-03-02 石药集团巨石生物制药有限公司 抗Nectin-4抗体、药物缀合物及其制备方法和用途

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