Winnonlin professional version

The pharmacokinetic parameters of PTX after a single intravenous or oral administration to rats were investigated by non-compartmental analysis using WinNonlin^® Professional version 5.2 software (Pharsight Corporation, Mountain View, CA, USA). The pharmacokinetic parameters estimated from the plasma concentration-time profiles were as follows: the area under the plasma concentration-time curve from 0 h to the last sampling time (AUC_last), the area under the plasma concentration-time curve from 0 h to infinity (AUC_inf), elimination half-life (t_1/2), elimination rate constant (K_e), apparent volume of distribution (V_d), total clearance (Cl_t), apparent volume of distribution following oral administration (V_d/F) and oral clearance (Cl_t/F). The maximum plasma concentration (C_max) and the time required to reach C_max (T_max) were measured directly from the concentration-time data.
The absolute bioavailability (AB, %) of PTX was calculated by the following equation:
$\text{AB}(\%)=\frac{\text{AUC}\hspace{0.17em}\text{po}/\text{Dose}\hspace{0.17em}\text{po}}{\text{AUC}\hspace{0.17em}\text{iv}/\text{Dose}\hspace{0.17em}\text{iv}}\times 100$
The relative bioavailability (RB, %) of PTX was calculated by the following equation:
$\text{RB}(\%)=\frac{\text{AUC}\hspace{0.17em}\text{co}–\text{admin}}{\text{AUC}\hspace{0.17em}\text{oral}\hspace{0.17em}\text{control}}\times 100$
, where AUC_{oral control} is the AUC obtained from the oral administration of PTX alone, and AUC_co-admin is the AUC obtained from the oral co-administration of PTX with VER, AC-603, or AC-786.

Pharmacokinetic parameters were calculated using noncompartmental methods (WinNonlin Professional, version 8.1; Pharsight Corporation, Cary, NC, USA) and included area under the plasma concentration–time curve from time zero to the last measurable time point (AUC_0–t) and from time zero to infinity (AUC_0–∞), terminal elimination half-life (t_1/2), peak plasma concentration (C_max), and the time to reach maximum plasma concentration (T_max). Both absolute (ratio of AUC_0–∞ for IN-MDZ or BC-MDZ to IV-MDZ) and relative (ratio of AUC_0–∞ for IN-MDZ to BC-MDZ) bioavailability of MDZ and 1-OH MDZ were determined, with the comparator value set at 100%.
To evaluate pharmacokinetics between IN-MDZ 2.5 mg and IV-MDZ 2.5 mg, an analysis of variance (ANOVA) with fixed effect (sequence, treatment, and period) and random effect (participant nested within the sequence) was performed on log-transformed C_max data to obtain the geometric least-squares mean (GLSM) for each treatment. The ratio of GLSM between the IN-MDZ 2.5 mg and IV-MDZ 2.5 mg was calculated.

Safety analyses were conducted with descriptive statistics (i.e., number of subjects, mean, SD, median, minimum, and maximum) for continuous variables and frequency and percentage for discrete variables (SAS version 9.4; SAS Institute, Cary, NC). PK analyses were conducted using noncompartmental methods (WinNonlin Professional version 6.4; Pharsight, Mountain View, CA). ECG analysis was based on the central tendency of ECG parameter changes from baseline. A categorical analysis was used for outliers. A morphological analysis was conducted for ECG waveform interpretation. The relationships between ETC‐206 concentrations and changes from baseline of ECG‐corrected QT Fridericia's formula (QTcF) were analyzed with a linear mixed effect modeling approach (Central Laboratory ERT, Philadelphia, PA; details are provided in Supplementary Material, TableS6). The exposure–response analysis including the calculation of the partial AUC between 0 and 4 hours postdose and the correlation between the AUC of ETC‐206 concentrations and the AUC of p‐eIF4E levels in PBMCs was conducted using SAS version 9.4.