Afuco™ Anti-Human CD44 ADCC Recombinant Antibody (RG7356), ADCC Enhanced (CAT#: AFC-122CL)

Figure 1 Pharmacokinetic assessments.

RG7356 serum concentration versus time plot in patients in Arm A who received the biweekly (q2w) (A) and weekly (qw) (B) dosing schedules

Menke-van der Houven,C.W.,van Oordt,C.G.R.,van Herpen, C.,Coveler, A. L.,Mahalingam, D., Verheul, H. M.,& Cannarile, M. A.(2016).First-in-human phase I clinical trial of RG7356,an anti-CD44 humanized antibody,in patients with advanced, CD44-expressing solid tumors.Oncotarget,7(48),80046.

Figure 2 Waterfall plot showing best response of individual patients according to RECIST criteria.

Of 61 evaluable patients, 21% (13/61) had stable disease (SD) as best response 45 patients had tumor lesion evaluation during the study. Abbreviations: PD (progressive disease); SD (stable disease).

Menke-van der Houven,C.W.,van Oordt,C.G.R.,van Herpen, C.,Coveler, A. L.,Mahalingam, D., Verheul, H. M.,& Cannarile, M. A.(2016).First-in-human phase I clinical trial of RG7356,an anti-CD44 humanized antibody,in patients with advanced, CD44-expressing solid tumors.Oncotarget,7(48),80046.

Figure 3 RG7356 induces a temporary reduction from baseline of CD14+ peripheral blood monocytes after first infusion.

(A),does not seem to have an effect on macrophage tumor infiltration (B), and generates a temporary release in cytokines. The latter seems to be independent of dose and/or dose-schedules (C-F). Dose-limiting toxicities (DLTs) occurred in 3 patients during the study: grade 3 febrile neutropenia on study day 18 and 2 cases of headache on study days 4 and 8 (C-F). Abbreviations: C (cycle); D (day); EOI (end of infusion); IL (interleukin); MCP-1 (monocyte chemoattractant protein-1); q2w (biweekly). A. Plot of mean changes from baseline of percent of CD14+ (± standard deviation) at RG7356 doses ≥300 mg for the q2w schedule. B. Waterfall plot of percent change in CD68+ macrophages. C.-F. Individual cytokine profiles.

Menke-van der Houven,C.W.,van Oordt,C.G.R.,van Herpen, C.,Coveler, A. L.,Mahalingam, D., Verheul, H. M.,& Cannarile, M. A.(2016).First-in-human phase I clinical trial of RG7356,an anti-CD44 humanized antibody,in patients with advanced, CD44-expressing solid tumors.Oncotarget,7(48),80046.

Figure 3 RG7356 induces a temporary reduction from baseline of CD14+ peripheral blood monocytes after first infusion.

The latter seems to be independent of dose and/or dose-schedules (C-D). Dose-limiting toxicities (DLTs) occurred in 3 patients during the study: grade 3 febrile neutropenia on study day 18 and 2 cases of headache on study days 4 and 8 (C-F). Abbreviations: C (cycle); D (day); EOI (end of infusion); IL (interleukin); MCP-1 (monocyte chemoattractant protein-1); q2w (biweekly). A. Plot of mean changes from baseline of percent of CD14+ (± standard deviation) at RG7356 doses ≥300 mg for the q2w schedule. B. Waterfall plot of percent change in CD68+ macrophages. C.-F. Individual cytokine profiles.

Menke-van der Houven,C.W.,van Oordt,C.G.R.,van Herpen, C.,Coveler, A. L.,Mahalingam, D., Verheul, H. M.,& Cannarile, M. A.(2016).First-in-human phase I clinical trial of RG7356,an anti-CD44 humanized antibody,in patients with advanced, CD44-expressing solid tumors.Oncotarget,7(48),80046.

Figure 4 A low dose of 25 μg ⁸⁹Zr-RG7356 was injected and the biodistribution of the radiolabeled antibody was evaluated at 1, 2, 3, and 6 d after injection.

Biodistribution of intravenously injected 89Zr-RG7356 (total protein dose: 25 μg) in CD44+ MDA-MB-231 (A) and CD44- HepG2(B)xenograft-bearing mice at 1,2,3,and 6 d after injection.Data are presented as %ID/g ± SD.

Menke-van der Houven,C.W.,van Oordt,C.G.R.,van Herpen, C.,Coveler, A. L.,Mahalingam, D., Verheul, H. M.,& Cannarile, M. A.(2016).First-in-human phase I clinical trial of RG7356,an anti-CD44 humanized antibody,in patients with advanced, CD44-expressing solid tumors.Oncotarget,7(48),80046.

Figure 5 Biodistribution of the mAb was assessed at 2, 3, and 6 d post injection and is summarized.

Dose escalation study of 89Zr-RG7356 in MDA-MB-231 xenograft bearing nude mice at 2 d (A), 3 d (B),and 6 d (C) after injection.A total mAb dose of 25, 50,200,500 and 1000μg (latter predose) was injected and data are presented as %ID/g ± SD.

Vugts, D. J., Heuveling, D. A., Stigter-van Walsum, M., Weigand, S., Bergstrom, M., van Dongen, G. A., & Nayak, T. K. (2014, March). Preclinical evaluation of 89Zr-labeled anti-CD44 monoclonal antibody RG7356 in mice and cynomolgus monkeys: prelude to phase 1 clinical studies. In MAbs (Vol. 6, No. 2, pp. 567-575). Taylor & Francis.

Figure 6 Immuno-PET studies in normal monkeys with ⁸⁹Zr-RG7356 revealed high uptake in the liver, spleen and the bone marrow.

Tissue to blood ratios at 2 d(A) and 5 d(B) after injection of 89Zr-RG7356 in cynomolgus macaques injected with 15 MBq 89Zr-RG7356 and 0.5,2,5,or 20 mg/kg unlabeled RG7356.

Vugts, D. J., Heuveling, D. A., Stigter-van Walsum, M., Weigand, S., Bergstrom, M., van Dongen, G. A., & Nayak, T. K. (2014, March). Preclinical evaluation of 89Zr-labeled anti-CD44 monoclonal antibody RG7356 in mice and cynomolgus monkeys: prelude to phase 1 clinical studies. In MAbs (Vol. 6, No. 2, pp. 567-575). Taylor & Francis.

Figure 7 Immuno-PET studies in normal monkeys with 89Zr-RG7356 revealed high uptake in the liver, spleen and the bone marrow

Biodistribution of 89Zr-labeled RG7356 in CD44+ MDA-MB-231 (responsive to therapy)and PL45 (non-responsive to therapy)tumor xenograft bearing mice at 1d (A)and 3 d(B)after injection.

Vugts, D. J., Heuveling, D. A., Stigter-van Walsum, M., Weigand, S., Bergstrom, M., van Dongen, G. A., & Nayak, T. K. (2014, March). Preclinical evaluation of 89Zr-labeled anti-CD44 monoclonal antibody RG7356 in mice and cynomolgus monkeys: prelude to phase 1 clinical studies. In MAbs (Vol. 6, No. 2, pp. 567-575). Taylor & Francis.

Figure 8 Maximum intensity projections of cynomolgus macaques injected with 15 MBq ⁸⁹Zr-RG7356 (0.1 mg/kg) and unlabeled RG7356. PET images were acquired 2 and 5 d after injection.

Vugts, D. J., Heuveling, D. A., Stigter-van Walsum, M., Weigand, S., Bergstrom, M., van Dongen, G. A., & Nayak, T. K. (2014, March). Preclinical evaluation of 89Zr-labeled anti-CD44 monoclonal antibody RG7356 in mice and cynomolgus monkeys: prelude to phase 1 clinical studies. In MAbs (Vol. 6, No. 2, pp. 567-575). Taylor & Francis.

Figure 9 The pharmacokinetic (PK) data for cycle 1 demonstrated that the time to peak concentration (tmax) occurred shortly after the end of the infusion (3–6 hours) in all cohorts.

For the every-other-week regimen, there was a supraproportional increase in mean exposure (Cmax and area under the curve [AUC]) from the 300-mg to 1200-mg dose and less than dose proportional from 1200-mg to 2400-mg dose. Total clearance (Cl) and volume of distribution (Vd) were high (relative to other IgG antibodies) at 300 mg, declined with increasing dose, and plateaued at 1200 mg, at which point target-mediated drug disposition (TMDD) saturation occurred. A similar PK profile was observed following the weekly regimen at the same dose. Mean half-life (t½) was 2–3 days and remained the same over the entire dose range.

Vugts, D. J., Heuveling, D. A., Stigter-van Walsum, M., Weigand, S., Bergstrom, M., van Dongen, G. A., & Nayak, T. K. (2014, March). Preclinical evaluation of 89Zr-labeled anti-CD44 monoclonal antibody RG7356 in mice and cynomolgus monkeys: prelude to phase 1 clinical studies. In MAbs (Vol. 6, No. 2, pp. 567-575). Taylor & Francis.

Figure 10 Leukemic stem cell (LSC) differentiation during treatment with RG7356.

LSC differentiation during treatment with RG7356 (patient 3015, 600 mg, twice weekly dose regimen, on treatment for 26 cycles). LSC differentiation is shown by reduction of percentage of CD34+ blasts and percentage increase of CD34−/CD38+ blasts in the bone marrow. Hematologic improvement (HI) is shown by absolute neutrophil count (ANC) increase (green line).

Vugts, D. J., Heuveling, D. A., Stigter-van Walsum, M., Weigand, S., Bergstrom, M., van Dongen, G. A., & Nayak, T. K. (2014, March). Preclinical evaluation of 89Zr-labeled anti-CD44 monoclonal antibody RG7356 in mice and cynomolgus monkeys: prelude to phase 1 clinical studies. In MAbs (Vol. 6, No. 2, pp. 567-575). Taylor & Francis.

Figure 11 Macrophage and leukemic stem cell changes during treatment with RG7356.

A. Macrophage (CD68) frequency and B. leukemic stem cell (CD34) reduction in the bone marrow was shown by immunohistochemistry analysis at baseline (pre-dose cycle 1, day 1 [C1D1]) and on treatment (pre-dose cycle 3, day 1 [C3D1]).

Vugts, D. J., Heuveling, D. A., Stigter-van Walsum, M., Weigand, S., Bergstrom, M., van Dongen, G. A., & Nayak, T. K. (2014, March). Preclinical evaluation of 89Zr-labeled anti-CD44 monoclonal antibody RG7356 in mice and cynomolgus monkeys: prelude to phase 1 clinical studies. In MAbs (Vol. 6, No. 2, pp. 567-575). Taylor & Francis.