NeuroMab™ Anti-EGFR BBB Shuttle Antibody(NRZP-1022-ZP3837)

Figure 1. Effect of 225 on the growth of established A431 tumor xenografts in nude mice. 107 cells were injected in the flank of the animals. Treatment started when tumors reached a mean volume of 2-300 mm3 and consisted of PBS or 1 mg/animal of 225 twice weekly for 5 weeks.

Figure 2. Effect of 225 and chimeric 225 (C225) on the growth of established A431 tumor xenografts in nude mice. Animals were treated with 1 mg/mouse of PBS twice a week for 5 weeks.

A: mean tumor volume; B: response index. The apparent tumor regression in the PBS control group at day 37 was due to the death of 3 out of 10 animals in the group at this time and was accompanied by a reduction in total tumor volume.

Figure 3 Effect of C225 on the growth of established A431 nude mouse xenografts. Animals were treated with 1 mg C225 or PBS twice weekly for 5 weeks.

The mean tumor volume of the C225 group showed a statistically significant biological effect compared to the control group (see text) A: mean tumor volume (asterisk indicates statistical significance relative to the control group); B: Relief index.

Figure 4. Dose response of C225 to established A431 xenograft growth in nude mice. Animals were treated with PBS, 1, 0.5 or 0.25 mg/animal twice weekly for 5 weeks.

Animals treated with 1 mg/dose of C225 showed statistically significant biological effects compared to controls (see text). A: Mean tumor volume (asterisk defines statistical significance relative to control); B: Response index. The decrease in RI at the 250 ug dose on day 47 was due to tumor reappearance in apparently tumor-free animals.

Figure 5 Inhibitory effect of C225 and heavy chain CDR-1 and heavy chain CDR-2 of monoclonal antibody 225 on A431 cells.

Figure 6. The relationship between the inhibitory effect of C225-doxorubicin conjugates on A431 cells in vivo and the concentration.

Figure 7 FACS Analysis of EGFR Expression on Human Prostate Cancer Cell Lines.

LNCaP (human prostate carcinoma, androgen-dependent), DU 145 and PC-3 (human prostate carcinoma, androgen-independent) and A431 (human epidermoid carcinoma) cells were removed from growth flasks with EDTA and stained with C225. Data are presented as MFI (Mean Fluorescence Intensity), an indirect measure of antigen expression. The results shown in this figure are representative of at least 5 experiments.

Figure 8. Inhibition of EGF-induced EGFR phosphorylation by C225 LNCaP, DU 145 and PC-3 monolayers were stimulated with EGF in the presence or absence of C225.

Cells were lysed, subjected to SDS PAGE, blotted, and screened with a mouse monoclonal antibody against PTyr (UBI, Lake Placid). Lane A: No addition (basal level of EGFR phosphorylation); Lane B: Stimulation of EGFR with 10 ng/ml EGF for 15 minutes at room temperature in the absence of C225; Lane C: In the presence of 10ug/ml C225 EGFR is stimulated with EGF.

Figure 9 Growth inhibition of established DU 145 xenografts by C225.

One million DU 145 cells in Matrigel were inoculated into nude mice (male, nu/nu). After tumors reached a mean volume of approximately 100 mm3 (day 20), animals were randomized (10 animals per group) and treated with PBS (control) or C225 (0.5 mg/dose, 10×). Animals were treated for 35 days and then followed up for 3 weeks. Mice without tumors or with small tumors were maintained for an additional 3 months. Significance was determined by Student's T-test, and p-values <0.5 were considered significant. A: average tumor volume; B: growth characteristics of tumors in PBS group; C: growth characteristics of tumors in C2-25 treatment group.

Figure 10. Effect of C225 on tumor elimination and survival.

Complete tumor elimination during the study was defined by the Response Index (RI). Animal mortality during the study period was considered treatment failure and included in the analysis. A: Remission index; B: Survival curve.