N8-GP in vitro functional data
| Parameters . | N8-GP . | FVIII* . |
|---|---|---|
| Kd of FVIIIa binding to FIXa (n = 12) | 0.94 ± 0.13 nM | 0.76 ± 0.07 nM |
| Km for FX activation (n = 10) | 4.9 ± 0.1 nM | 4.8 ± 0.1 nM |
| kcat for FX activation (n = 10) | 5.0 ± 0.4 s−1 | 5.5 ± 1.0 s−1 |
| kapp for thrombin-mediated FVIII activation | ||
| Without VWF (n = 10-12) | 7.5 ± 1.0 × 10−3 min−1 | 10.8 ± 1.0 × 10−3 min−1 |
| With VWF (n = 6) | 17 ± 3 × 10−3 min−1 | 35 ± 5 × 10−3 min−1 |
| Rate of FVIIIa inactivation by APCs (n = 10) | 0.22 ± 0.01 min−1 | 0.23 ± 0.01 min−1 |
| Kd for binding to LRP | 13 nM | 4.1 nM |
| On-rate for LRP binding | 1.3 × 105 M s−1 | 4.1 × 105 M s−1 |
| Off-rate for LRP binding | 0.00170 × 105 s−1 | 0.00168 × 105 s−1 |
| Stability in plasma, t1/2 | 195 h (177-213 h) | 210 h (185-236 h) |
| Parameters . | N8-GP . | FVIII* . |
|---|---|---|
| Kd of FVIIIa binding to FIXa (n = 12) | 0.94 ± 0.13 nM | 0.76 ± 0.07 nM |
| Km for FX activation (n = 10) | 4.9 ± 0.1 nM | 4.8 ± 0.1 nM |
| kcat for FX activation (n = 10) | 5.0 ± 0.4 s−1 | 5.5 ± 1.0 s−1 |
| kapp for thrombin-mediated FVIII activation | ||
| Without VWF (n = 10-12) | 7.5 ± 1.0 × 10−3 min−1 | 10.8 ± 1.0 × 10−3 min−1 |
| With VWF (n = 6) | 17 ± 3 × 10−3 min−1 | 35 ± 5 × 10−3 min−1 |
| Rate of FVIIIa inactivation by APCs (n = 10) | 0.22 ± 0.01 min−1 | 0.23 ± 0.01 min−1 |
| Kd for binding to LRP | 13 nM | 4.1 nM |
| On-rate for LRP binding | 1.3 × 105 M s−1 | 4.1 × 105 M s−1 |
| Off-rate for LRP binding | 0.00170 × 105 s−1 | 0.00168 × 105 s−1 |
| Stability in plasma, t1/2 | 195 h (177-213 h) | 210 h (185-236 h) |
The FVIII used was Advate except for binding to LRP and stability in plasma, where turoctocog alfa was used. Data are mean and standard deviation of the indicated number of experiments. For LRP binding, values are based on fit of data obtained with 12.5 nM, 3.13 nM, 0.78 nM, 0.2 nM (see Figure 2), and 0.05 nM FVIII. The t1/2 for stability in plasma is calculated by linear regression of 8 data points distributed evenly above and below 50% activity. The values shown are best-fit values and 95% confidence intervals as reported by the least-squares software (Graph Pad Prism).