Safety, Tolerability and Pharmacokinetic Profiles of MOTREM (LR12) in Healthy Male Subjects

The number of subjects experiencing treatment emergent adverse events was collected to assess the safety and tolerability of MOTREM (LR12) in comparison with placebo.

Time frame: 30-44 days

Plasma concentrations of LR12 were measured by a validated liquid chromatography-mass spectrometry (LC-MS/MS) assay and analyzed using noncompartmental methods to obtain estimates of the pharmacokinetic parameter of Maximum Plasma Concentration (Cmax).

Time frame: Maximum Plasma Concentration (Cmax) was assessed only for Groups 3-8 who received loading dose over 15 min and maintenance dose over 7 h and 45 min. Cmax is determined over a period of time starting from predose to 10h after start of the loading dose.

Population: In cohort 1 and cohort 2, there was 1 subject per cohort and the subjects received only a loading dose. LR12 plasma concentrations were quantifiable only at 15 min post-start of the infusion. For this reason, no PK analysis was performed for these two cohorts and for this reason results were not presented.

Plasma concentrations of LR12 were measured by a validated LC-MS/MS assay and analyzed using noncompartmental methods to obtain estimates of the PK parameter of AUC0-∞. AUC0-∞ represents the area under the plasma concentration-time curve (h.ng/mL) from time 0 to infinity (AUC0- ∞= AUC0-t + \[Ct/ke\], where Ct = the observed concentration of drug for the last sample on the PK profile in which drug was detected, and ke represents the terminal rate constant). The percentage of extrapolation of AUC0-∞ should not exceed 20%.

Time frame: AUC0-∞ was assessed only for Groups 3-8 who received loading dose over 15 min and maintenance dose over 7 h and 45 min. AUC0-∞ is determined over a period of time starting time zero (predose) to infinity (cf. extrapolation formula in the above section).

Population: The percentage of extrapolation of AUC0-∞ was below 1% in subjects up to cohort 5, and due to this it was not determined for these subjects. For this reason, results were not presented for cohorts 1-5.

Plasma concentrations of LR12 were measured by a validated LC-MS/MS assay and analyzed using noncompartmental methods to obtain estimates of the PK parameter of the area under the plasma concentration curve (h.ng/mL) from time zero (predose) to the time of last observed concentration (t) measured (which can go up to 10h post loading dose start) using a linear trapezoidal method (AUC0-t).

Time frame: AUC0-t was assessed only for Groups 3-8 who received loading dose over 15 min and maintenance dose over 7 h and 45 min. AUC0-t is determined over a period of time starting time zero (predose) to the time of last observed concentration (t).

Population: There was 1 subject per cohort in cohort 1 and cohort 2 and the subjects received only a loading dose. LR12 (nangibotide) plasma concentrations were quantifiable only at 15 min post-start of the infusion. For this reason, no PK analysis was planned for these two cohorts and no results were presented.

Plasma concentrations of LR12 were measured by a validated LC-MS/MS assay and analyzed using noncompartmental methods to obtain estimates of the PK parameter of systemic clearance (CL). The systemic clearance was estimated using the formula: CL = K0/ Css where K0 is the perfusion rate (ng/kg/h) and Css is the concentration at the end of perfusion (C at 465 min). Of note, this narrow range of the clearance values indicates that the apparent increases in t1/2 and volume of distribution as a function of doses are not due to a non-linearity in the pharmacokinetics, but to the limit of quantification of the bioanalytical assay.

Time frame: CL was assessed only for Groups 3-8 who received loading dose over 15 min and maintenance dose over 7 h and 45 min (465 min). CL is calculated based on the perfusion rate (ng/kg/h) and the concentration at the end of perfusion (7h45min = 465min).

Population: There was 1 subject per cohort and the subjects received only a loading dose in cohort 1 and cohort 2. LR12 (nangibotide) plasma concentrations were quantifiable only at 15 min post-start of the infusion. No PK analysis was planned for these two cohorts. Also, CL values for cohort 3 were non-calculable. For this reason, results for cohorts 1-3 are not presented.

Plasma concentrations of LR12 were measured by a validated LC-MS/MS assay and analyzed using noncompartmental methods to obtain estimates of the PK parameter of steady state concentration during the maintenance infusion (Cavg30-465).

Time frame: Cavg30-465 was assessed only for Groups 3-8 who received loading dose over 15 min and maintenance dose over 7 h and 45 min. Cavg30-465 is determined over a period of time starting from predose to 10h after start of the loading dose.

Population: In cohort 1 and cohort 2, there was 1 subject per cohort and the subjects received only a loading dose. LR12 plasma concentrations were quantifiable only at 15 min post-start of the infusion. For this reason, no PK analysis was performed for these two cohorts. For Cohort 3 values for Cavg30-465 were non-calculable. Hence, the results for Cohorts 1,2 and 3 are not presented.

Plasma concentrations of LR12 were measured by a validated LC-MS/MS assay and analyzed using noncompartmental methods to obtain estimates of the PK parameter of terminal half-life (t1/2). Of note, apparent increase in half-life at increasing doses is explained by the detection of a slow elimination phase, which was below limit of quantification for the lower doses.

Time frame: t1/2 was assessed only for Groups 3-8 who received loading dose over 15 min and maintenance dose over 7 h and 45 min. t1/2 is determined over a period of time starting from 7 h and 45 min to 10h after start of the loading dose (decaying period).

Population: Up to cohort 5, due to paucity of the data, t1/2 could not be determined, hence the results for these cohorts are not presented.

This PK parameter was calculated from measured plasma concentrations of LR12 and it represents the time of last observed concentration.

Time frame: t last was assessed only for Groups 3-8 who received loading dose over 15 min and maintenance dose over 7 h and 45 min. t last is determined as the time of last observed concentration which can go up to 10h after loading dose start.

Population: There was 1 subject per cohort in cohort 1 and 2 and the subjects received only a loading dose. LR12 plasma concentrations were quantifiable only at 15 min post-start of the infusion. No PK analysis was planned for these two cohorts. For this reason, results for cohort 1 and 2 were not presented.

Plasma concentrations of LR12 were measured by a validated LC-MS/MS assay and analysed using noncompartmental methods to obtain estimates of the PK parameter of the time at which Cmax is apparent, identified by inspection of the plasma drug concentration vs.time data by WinNonlin (tmax).

Time frame: tmax was assessed only for Groups 3-8 who received loading dose over 15 min and maintenance dose over 7 h and 45 min. tmax is determined over a period of time starting from predose to 10h after start of the loading dose.

Population: There was 1 subject per cohort in cohort 1 and 2 and the subjects received only a loading dose. LR12 plasma concentrations were quantifiable only at 15 min post-start of the infusion. No PK analysis was planned for these two cohorts. For this reason, results for cohort 1 and 2 were not presented.

The volume of distribution was estimated according to the following equation: V= CL x MRT (mean residence time). However, regarding administration procedures, MRT could not be calculated precisely. Furthermore, in some cases MRT could not be estimated. Thus, the following formula was used V = CL / ke. The terminal rate constant (ke) was estimated by log-linear regression analysis on data points visually assessed to be on the terminal log-linear phase. Of note, it can be said that this apparent increase in V is also explained by the detection of a slow elimination phase, which was below limit of quantification for the lower doses.

Time frame: V was assessed only for Groups 3-8 who received loading dose over 15 min and maintenance dose over 7 h and 45 min. V is derived from both CL and ke which are calculated from the LR12 concentration time curve from predose to 10h after loading dose start.

Population: There was 1 subject per cohort In cohort 1 and cohort 2 and the subjects received only a loading dose. LR12 plasma concentrations were quantifiable only at 15 min post-start of the infusion. No PK analysis was planned for these two cohorts. Also, V values for cohort 3, 4 and 5 were non-calculable. For this reason, no results were presented for cohorts 1-6.