The in vitro complement inhibitory activity of CG001. (A-C) The complement inhibitory effect of CG001 in NHS. The complement was activated via (A) CP, (B) LP, or (C) AP, and CG001 was compared with eculizumab as a control. (A) IC50 = 4.71 ± 0.28 nM (CG001) or 1.54 ± 0.14 nM (eculizumab) for CP, IC95 = 510.95 ± 91.98 nM (CG001) or 5.5 ± 0.29 nM (eculizumab) for CP; (B) 2.52 ± 0.62 nM (CG001) or 1.02 ± 0.10 nM (eculizumab) for LP, and IC95 was 365.73 ± 114.62 nM (CG001) or 5.71 ± 0.48 nM (eculizumab) for LP; and (C) 0.83 ± 0.12 nM (CG001) or 8.56 ± 0.24 nM (eculizumab) for AP and IC95 was 4.0 ± 0.76 nM (CG001) or 22.0 ± 0.93 nM (eculizumab) for AP. (D-E) The inhibitory effect of CG001 on complement component-depleted human serum. (D) IC50 = 10.932 ± 1.629 nM in FB-depleted human serum selective for CP activity, and (E) IC50 = 0.858 ± 0.034 nM in C4-depleted human serum selective for AP activity. (F-I) The complement inhibitory effect of CG001 in normal monkey serum (NMS) and normal rat serum (NRS). (F) IC50 = 9.335 ± 2.328 nM for CP in NMS, (G) 0.455 ± 0.037 nM for AP in NMS, (H) IC50 = 192.5 ± 36.2 nM for CP in NRS, or (I) 96.0 ± 6.8 nM for AP in NRS. Complement activities in human and monkey sera were measured by the Wieslab complement system kit for panels A-G. To evaluate the complement activities in rat serum, (H) CP was activated by 0.2% sheep erythrocyte hemolysin and 1% NRS to induce hemolysis of sRBCs, whereas (I) AP was directly activated by 18% NHS to induce hemolysis of rRBCs. Values represent mean ± SD, and experiments were performed in triplicate.
