Article Figures & Data
Figures
- Fig. 1.
Mean (±S.D.) concentrations of radioactivity in blood and plasma and mean (±S.D.) concentrations of parent compound (LY3202626) after administration of a single 10-mg (100 µCi) oral dose of [14C]LY3202626 to healthy subjects.
- Fig. 2.
Mean (+S.D.) cumulative percentage of radioactive dose recovered in urine and feces at specified intervals after administration of a single 10-mg (100 µCi) oral dose of [14C]LY3202626 to healthy subjects.
- Fig. 3.
UV chromatogram (250 nm) generated from injection of diluted plasma spiked with synthetic standards of M1, M2, M3, M4, M5, M16, and P (parent) overlaid against a 14C radiochromatogram based on the AMS detection of fractions and fraction pools collected from direct injection of 0- to 24-hour AUC pool plasma after administration of a single 10-mg (100 µCi) oral dose of [14C]LY3202626 to healthy subjects. Note that UV trace of the parent (P) is obscured by its radioactive trace.
- Fig. 4.
Reconstructed radioprofiles of urine after administration of a single 10-mg (100 µCi) oral dose of [14C]LY3202626 to healthy subjects. P, parent,
- Fig. 5.
Reconstructed radioprofiles of feces after administration of a single 10-mg (100 µCi) oral dose of [14C]LY3202626 to healthy subjects. P, parent.
- Fig. 6.
(A) Extracted ion chromatograms of LY3202626 and its dimedone adduct after incubation of LY3202626 in human liver microsomes in the absence (A) and presence of NADPH (B). (B) The proposed metabolic pathway for the formation of LY3202626-dimedone adduct after incubation of LY32026 in human liver microsome incubations in the presence of NADPH and dimedone.
- Fig. 7.
Extracted ion chromatograms of M2 (LSN3207841) and incubations under anaerobic condition for 168 hours with brain heart infusion medium (A), with deactivated human fecal microflora (B) and non-deactivated human fecal microflora (C).
- Fig. 8.
Extracted ion chromatograms of M16 (LSN3420637) incubations in human hepatocytes for 4 hours in the absence of any inhibitor (A), in the presence of ABT (B), and in the presence of hydralazine (C).
- Fig. 9.
The proposed metabolic scheme for LY3202626 after a single 10-mg oral dose of [14C]LY3202626 to healthy humans. This test article is dual radiolabeled, and the dosing included 1:1 mix of both labeled compounds; * signifies the positions of the radiolabel. F, P, and U indicate presence in feces, plasma and urine, respectively.
- Fig. 10.
Hypothesis illustrating enterohepatic recirculation of radioactive material and possible cause of slow excretion of radioactivity, involving microbial reduction of metabolite M2 to M16 in the gut and reabsorption of M16, followed by hepatic oxidation of M16 back to M2.
Tables
- TABLE 1
Summary of pharmacokinetic parameter estimates of LY3202626 in plasma and total radioactivity in plasma and whole blood after oral administration of a single 10-mg dose of [14C]LY3202626
Concentrations of total radioactivity in whole blood were not quantifiable for a sufficient duration to reliably determine AUC(0–∞). Instead, partial AUCs up to the last common quantifiable time point for all subjects in plasma and whole blood, AUC(0–48), and AUC(0–10), respectively, were used for calculation of ratios of plasma LY3202626 relative to plasma total radioactivity and ratios of whole blood to plasma total radioactivity.
PK Parameters Geometric Mean (CV%) Plasma LY3202626 (N = 6) Plasma Total Radioactivitya (N = 6) Whole Blood Total Radioactivitya (N = 6) AUC(0–tlast) (ng·h/ml) 294 (48) 814 (31) 210 (60) AUC(0–∞) (ng·h/ml) 298 (47) 1120 (27) NC %AUC(tlast–∞) 1.19 (48) 27.0 (19) NC AUC(0–10) (ng·h/ml) 70.0 (36) 207 (14) 146 (17) AUC(0–48) (ng·h/ml) 227 (37) 711 (15) NC Cmax (ng/ml) 10.7 (32) 25.2 (16) 21.0 (18) tmaxbb (h) 4.00 (2.00–10.00) 3.50 (2.00–5.00) 4.00 (2.00–4.05) t1/2c (h) 24.5 (17.0–52.2) 32.5 (25.1–45.4) NC CL/F (l/h) 33.6 (47) NA NA Vz/F (l) 1190 (19) NA NA Vss/F (l) 1140 (28) NA NA - TABLE 2
Percent of sample radioactivity as [14C]LY3202626 or metabolites of [14C]LY3202626 in pooled human plasma samples after a single oral dose of 10 mg of [14C]LY3202626
Peak Protonated Molecular ion [M + H]+ Proposed Metabolite Identification Percent of Radioactivity (% of Run) 4 hour 144 hour 288 hour AUC Pool (0–24 hours) Parent 499 LY3202626 43.4 8.96 1.63 38 M1 363 Amide hydrolysis to the amine (aniline) (LSN3200635) 1.28 17.2 24.3 2.27 M2 485 O-desmethyl (LSN3207841) 4.13 11.3 10.3 4.41 M3 155 Amide hydrolysis to the carboxylic acid (LSN3170994) 3.64 <1.0 <1.0 3.86 M4 212 M3 + glycine (LSN3329581) 10.3 1.57 <1.0 11.9 M5 405 N-acetyl of M1 (LSN3226305) <0.5 <1.0 <1.0 <1.0 M16 460 M2 − O (reduction) (LSN3420637) <1.0 2.1 1.81 <1.0 Region A 30.0 46.9 54.3 33.5 Total 92.76 88.03 92.34 93.94 Total radioactivity recovery 103.0 101.3 103.0 96.7 - TABLE 3
Percent dose of LY3202626 and its metabolites excreted in urine and feces from healthy human subjects after a single oral dose of 10 mg of [14C]LY3202626 from 0 to 508 hours
Peak Protonated Molecular ion [M + H]+ Proposed Metabolite Identification Percent of Dose Urine Feces Total Parent 499 LY3202626 2.25 2.02 4.27 M1 363 Amide hydrolysis to the amine (aniline) (LSN3200635) 1.21 ND 1.21 M2 485 O-desmethyl (LSN3207841) 8.88 10.75 19.63 M4 212 M3 + glycine (LSN3329581) 21.27 ND 21.27 M5 405 N-acetyl of M1 (LSN3226305) 0.18 1.32 1.5 M16 460 M2 − O (reduction) (LSN3420637) ND 6.94 6.94 M21 499 Same mass as parent 1.34 1.55 2.89 Total % identified 35.15 22.57 57.72 Total % dose in the sample analyzed 40.99 28.81 69.8 - TABLE 4
Permeability in MDCK-MDR1 monolayers for LSN3207841(M2) and LSN3420637 (M16) and LSN3182170 (HCl salt of LY3202626, parent)
Compound rD0, µM Pe ± S.D. × 10−6 cm/s % Cell ± S.D. % Recovery Pe BL/AP Pe c/l P-gp P-gp Inhibition % Comments 5 µM 25 µM M2 P-gp inhibited Yes 0 0 Very slow Pe, very low % cell, inhibition unlikely A-B 3.5 1.1 ± 0.25 0.34 ± 0.00 102 ± 0.56 0.71 29 B-A 3.3 0.80 ± 0.00 0.31 ± 0.00 100 ± 5.1 P-gp active A-B 3.3 0.57 ± 0.05 0.30 ± 0.02 100 ± 1.4 21 B-A 3.4 12 ± 0.26 0.27 ± 0.02 89 ± 3.6 M16 P-gp inhibited Yes 1 21 Fast Pe, moderate % cell, inhibition unlikely A-B 3.5 53 ± 0.14 20 ± 0.18 154 ± 0.40 0.99 6.1 B-A 2.8 53 ± 0.04 19 ± 0.73 130 ± 3.9 P-gp active A-B 2.6 16 ± 2.1 11 ± 0.09 115 ± 5.7 6.0 B-A 2.6 97 ± 0.62 7.9 ± 0.42 125 ± 4.1 Parent P-gp inhibited No 6 25 Fast Pe, high partitioning, inhibition unlikely A-B 2.1 19 ± 2.3 50 ± 0.53 92 ± 8.6 1.2 1.5 B-A 2.4 21 ± 0.02 28 ± 0.00 97 ± 5.5 P-gp active A-B 2.8 15 ± 0.13 49 ± 1.6 88 ± 0.25 1.8 B-A 2.7 27 ± 1.1 27 ± 5.4 94 ± 1.4 rD0 is the measured concentration of recovered donor solution at t = 60 min. Pe is the apparent permeability coefficient (n = 2) with 60-min time interval: AB, BA>15, fast; AB, BA is 2–15, moderate; AB, BA is 1–2, slow; AB, BA is <1, very slow. % Cell is the amount extracted from cell with methanol relative to recovered mass: <1%, very low; 1%–5%, low; 6%–20%, moderate; 21%–89%, high; ≥90%, very high (invalidates Pe values because compound prefers lipid environment, assay artifact). %Recovery is the mass of D60 + cell60 + R60/D0 (measured concentration of donor solution at t = 0) × 100. If >100%, then D0 is underestimated. Pe BL/AP is the ratio of Pe values in B-to-A and A-to-B directions; >3.0 + active efflux; Pe C/I is the net efflux ratio = inhibited BA/AB and control BA/AB ratio. P-gp Yes (Y), No (N), or Unknown (U): if ≥3, Y; <3, N; if % Cell > 90, U. P-gp inhibition, two concentrations (5 and 25 µM) tested for calcein AM uptake assay in MDCK-MDR1 cells; % inhibition vs. LSN335984 (EC50 = 0.15–0.3 µM), pending at 5 µM: 0–20, inhibition unlikely, 21–50, partial inhibition possible, >51%, partial inhibition likely.
Data Supplement
- Supplemental Data -
Supplementary Methods
Supplementary Table 1 - Accurate Mass Values and Characteristic Product Ions for LY3202626 and Metabolites Observed in Rat, dog and Human.
Supplementary Table 2 - Effect of ABT and hydralazine on the formation of M2 following M16 (LSN3420637) incubation in hepatocytes for 4 hours.
Supplementary Table 3A - Relative Distribution of Radioactivity (%) Among Plasma Metabolites Identified in the Radioprofiles of Male Sprague Dawley Rat Plasma After a Single Oral Dose of 30 mg/kg [14C]LY3202626.
Supplementary Table 3B - Relative Distribution of Radioactivity (%) Among Plasma Metabolites Identified in the Radioprofile of Dog Plasma following a Single Intravenous Dose of 1 mg/kg [14C]LY3202626 and a Single Oral Dose of 10 mg/kg [14C]LY3202626 administration.
Supplementary Table 4 - Metabolites Identified in urine, bile and/or feces across rat, dog and human (% dose excretion).
Supplementary Figure 1A - Mean (± SD) cumulative percent of radioactive dose in urine and feces after a single 30 mg/kg oral administration of [14C]LY3202626 hydrate to intact and bile duct cannulated rats.
Supplementary Figure 1B - Mean (± SD) cumulative percent of radioactive dose in urine and feces after a single 10 mg/kg oral and 1 mg/kg intravenous administration of [14C]LY3202626 hydrate to dogs.
Supplementary References
- Supplemental Data -
