PDF file - 166K, Supplementary Table S1: TF HuMab: target binding characteristics and inhibition of FXa generation. Supplementary Table S2: Cross-competition between TF HuMab. Supplementary Table S3: Thromboelastography: coagulation of LPS-stimulated whole blood in the presence of 20 microg/mL TF-011, TF-013 or TF-098. Supplementary Figure S1: TF HuMab inhibit TF:FVIIa-induced ERK1/2 phosphorylation. Supplementary Figure S2: TF:FVIIa-induced IL-8 production in MDA-MB-231 cells is reduced in the presence of TF HuMab. Supplementary Figure S3: Binding of TF HuMab is unaltered after conjugation with vcMMAE or mcMMAF. Supplementary Figure S4: Anti-tumor activity of TF-ADCs in the A431 and HCT-116 xenograft models. Supplementary Figure S5: Anti-tumor activity of unconjugated TF HuMab in vivo. Supplementary Figure S6: Post-paclitaxel treatment with TF-011-MMAE in PDX models. Supplementary Figure S7: Full-length blots for Figure 1C.
ARTICLE ABSTRACT
Tissue factor (TF) is aberrantly expressed in solid cancers and is thought to contribute to disease progression through its procoagulant activity and its capacity to induce intracellular signaling in complex with factor VIIa (FVIIa). To explore the possibility of using tissue factor as a target for an antibody-drug conjugate (ADC), a panel of human tissue factor–specific antibodies (TF HuMab) was generated. Three tissue factor HuMab, that induced efficient inhibition of TF:FVIIa-dependent intracellular signaling, antibody-dependent cell-mediated cytotoxicity, and rapid target internalization, but had minimal impact on tissue factor procoagulant activity in vitro, were conjugated with the cytotoxic agents monomethyl auristatin E (MMAE) or monomethyl auristatin F (MMAF). Tissue factor–specific ADCs showed potent cytotoxicity in vitro and in vivo, which was dependent on tissue factor expression. TF-011-MMAE (HuMax-TF-ADC) was the most potent ADC, and the dominant mechanism of action in vivo was auristatin-mediated tumor cell killing. Importantly, TF-011-MMAE showed excellent antitumor activity in patient-derived xenograft (PDX) models with variable levels of tissue factor expression, derived from seven different solid cancers. Complete tumor regression was observed in all PDX models, including models that showed tissue factor expression in only 25% to 50% of the tumor cells. In conclusion, TF-011-MMAE is a promising novel antitumor agent with potent activity in xenograft models that represent the heterogeneity of human tumors, including heterogeneous target expression. Cancer Res; 74(4); 1214–26. ©2013 AACR.
