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W. J. Moree et al. / Bioorg. Med. Chem. Lett. 20 (2010) 2316–2320
a human microdose study.17 While clearance projections from allo-
metric scaling were reasonably predictive (predicted clearance:
13.5 mL/min/kg, measured mean clearance: 12.1 mL/min/kg), the
measured human half-life for 16d was longer than anticipated
(13 h vs the estimate of 1.6 h predicted from allometry) and longer
than for 2 (6.8 h). This discrepancy in half-life prediction was
attributed to both an underestimate of the volume of distribution
of 16d (Vss measured as 5.6 L/kg in humans16 versus 1.7 L/kg from
allometric scaling) and an overestimate of clearance. Even though a
small sample size was used in the microdose study, 16d showed an
increasing trend to high variability in exposure compared to 2 fol-
lowing both iv (hClsys, Table 4) and oral dosing (AUC0-t, Table 4).
PK variability in our leads was considered a significant issue be-
cause of the potential for varying exposures to adversely impact
duration of action. Although a specific mechanism for the higher
variability remains unclear, the higher clearance of 16d compared
to 2, coupled with lower bioavailability (23% vs 47% for 2) is con-
sistent with mechanistic expectations that reduced metabolic sta-
bility generally contributes to increased variation in PK profile.17
Based on these human PK data, neither compound 16d nor 2 was
pursued further. The more stable 28b may therefore represent a
more suitable backup to 2 and warrants further assessment.
In summary, discrete modifications around a lead benzothio-
phene 2 provided several compounds with a comparable in vitro
profile and a small improvement in hERG selectivity. From rat PK
studies, compounds with a range of projected human half-lives
were identified. Compound 16d was chosen as a representative
high clearance compound and assessed in a human microdose
study. In contrast to allometric projections, this compound had a
longer half-life in humans compared to 2. Unfortunately, both
compounds 2 and 16d showed a trend to high PK variability indi-
cating neither analog was suitable for further development. Com-
pound 28b, with improved metabolic stability, may represent the
more promising backup candidate.
Supplementary data
Supplementary data associated with this article can be found, in
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The authors wish to thank Michael Johns for permeability
determinations, John Harman, Shawn Ayube and Chris DeVore for
analytical support. Dr. Tao Hu and Mila Lagman for chiral separa-
tions and Dr. John Saunders, Dr. Paul Conlon, Dr. H. Bozigian, and
Dr. Wendell Wierenga for program support.