Figure 4.

PK/PD of hOTC-mRNA/LNP in wild-type mice

hOTC mRNA levels in the liver of female BALB/c mice intravenously injected with hOTC-mRNA/LNP at 1.0 or 2.5 mg/kg. Liver specimens were harvested 1, 2, 3, 5, and 7 days after the injection. Untreated (UT) mice were used as controls. (A) Time course changes of hOTC mRNA levels in the liver specimens. Values are expressed as means ± SD with individual values. n = 3 per each time. Fitting to one phase exponential equation in the case of 1.0 mg/kg (B) and 2.5 mg/kg (C). The equation is Y = (Y0 − Plateau)∗exp(−K∗X) + Plateau, where Y0 is the Y value when X is 0, Plateau is the Y value at infinite times, K is the rate constant, and the t_1/2 is computed as ln(2)/K. Values are expressed as means ± SD. Time course changes of hOTC protein levels in the liver of mice intravenously injected with hOTC-mRNA/LNP at 0.3 or 1.0 mg/kg. Liver samples were collected 1, 7, 14, and 21 days after the treatment. n = 3 per time. Western blot of hepatic hOTC protein expression of mice treated with hOTC-mRNA/LNP at 0.3 mg/kg (D) and 1.0 mg/kg (E). HSP90 proteins were used as an internal control. (F) Quantified hOTC protein levels revealed by western blot in (D) and (E). Values are expressed as means ± SD with individual values. (G) Fitting to plateau followed by one-phase exponential decay, combined with data of (F) and Figure S6B at 1.0 mg/kg. The equation is Y = IF(X < X0, Y0, Plateau + (Y0 − Plateau)∗exp(−K∗(X-X0))), where X0 is the time at which the decay begins, Y0 is the average Y value up to time X0, Plateau is the Y value at infinite times, and K is the rate constant). In this case, Plateau was constrained to a constant value of zero. The t_1/2 is computed as ln(2)/K. Values are expressed as means ± SD. n = 3 or 6 per time.