Development of tetravalent IgG1 dual targeting IGF-1R-EGFR antibodies with potent tumor inhibition

被引：40

作者：

Croasdale, Rebecca ^{[1
]}

Wartha, Katharina ^{[2
]}

Schanzer, Juergen M. ^{[1
]}

Kuenkele, Klaus-Peter ^{[2
]}

Ries, Carola ^{[2
]}

Mayer, Klaus ^{[1
]}

Gassner, Christian ^{[1
]}

Wagner, Martina ^{[1
]}

Dimoudis, Nikolaos ^{[1
]}

Herter, Sylvia ^{[3
]}

Jaeger, Christiane ^{[4
]}

Ferrara, Claudia ^{[4
]}

Hoffmann, Eike ^{[1
]}

Kling, Lothar ^{[1
]}

Lau, Wilma ^{[1
]}

Staack, Roland F. ^{[1
]}

Heinrich, Julia ^{[1
]}

Scheuer, Werner ^{[2
]}

Stracke, Jan ^{[1
]}

Gerdes, Christian ^{[3
]}

Brinkmann, Ulrich ^{[1
]}

Umana, Pablo ^{[3
,4
]}

Klein, Christian ^{[3
]}

机构：

[1] Roche Diagnost GmbH, Roche Pharma Res & Early Dev pRED, Large Mol Res, D-82372 Penzberg, Germany

[2] Roche Diagnost GmbH, Roche Pharma Res & Early Dev pRED, Discovery Oncol, D-82372 Penzberg, Germany

[3] Roche Glycart AG, Roche Pharma Res & Early Dev pRED, Discovery Oncol, CH-8952 Schlieren, Switzerland

[4] Roche Glycart AG, Roche Pharma Res & Early Dev pRED, Large Mol Res, CH-8952 Schlieren, Switzerland

来源：

ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS | 2012年 / 526卷 / 02期

关键词：

Antibody engineering; Bispecific antibodies; Single chain Fv; IGF-1R; EGFR; GA201; R1507; GROWTH-FACTOR RECEPTOR; FACTOR-I RECEPTOR; DISULFIDE-STABILIZED FV; CONSERVED FRAMEWORK REGIONS; HUMAN MONOCLONAL-ANTIBODY; HUMAN GLIOBLASTOMA CELLS; ADVANCED SOLID TUMORS; BISPECIFIC ANTIBODY; ANTITUMOR-ACTIVITY; CANCER-CELLS;

D O I：

10.1016/j.abb.2012.03.016

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

070307 [化学生物学]; 071010 [生物化学与分子生物学];

摘要：

In this study we present novel bispecific antibodies that simultaneously target the insulin-like growth factor receptor type I (IGF-1R) and epidermal growth factor receptor (EGFR). For this purpose disulfide stabilized scFv domains of the EGFR/ADCC antibody GA201 were fused via serine-glycine connectors to the C-terminus of the heavy (XGFR2) or light chain (XGFR4), or the N-termini of the light (XGFR5) or heavy chain (XGFR3) of the IGF-1R antibody R1507 as parental IgG1 antibody. The resulting bispecific IGF-1R-EGFR antibodies XGFR2, XGFR3 and XGFR4 were successfully generated with yields and stability comparable to conventional IgG1 antibodies. They effectively inhibited IGF-1R and EGFR phosphorylation and 3D proliferation of H322M and H460M2 tumor cells, induced strong down-modulation of IGF-1R as well as enhanced EGFR down-modulation compared to the parental EGFR antibody GA201 and were ADCC competent. The bispecific XGFR derivatives showed a strong format dependent influence of N- or C-terminal heavy and light chain scFv attachment on ADCC activity and an increase in receptor downregulation over the parental combination in vitro. XGFR2 and XGFR4 were selected for in vivo evaluation and showed potent anti-tumoral efficacy comparable to the combination of monospecific IGF-1R and EGFR antibodies in subcutaneous BxPC3 and H322M xenograft models. In summary, we have managed to overcome issues of stability and productivity of bispecific antibodies, discovered important antibody fusion protein design related differences on ADCC activity and receptor downmodulation and show that IGF-1R-EGFR antibodies represent an attractive therapeutic strategy to simultaneously target two key components de-regulated in multiple cancer types, with the ultimate goal to avoid the formation of resistance to therapy. (C) 2012 Elsevier Inc. All rights reserved.

引用

页码：206 / 218

页数：13

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