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Select compounds were periodically examined using bovine,
rat, and human sources of an active cardiac myosin fragment
to assess variability across species. No difference in activity
was observed across species.
(9)
Compound specificity with respect to muscle type was
evaluated by comparing the effect of the compound on the
actin-stimulated ATPase activity of a panel of myosin iso-
forms [cardiac (bovine), skeletal (rabbit), and smooth muscle
(bovine uterus or chicken gizzard)] at a single dose of 40, 50,
or 100 μM compound. The data for compound 1 are shown in
Figure S1 in the Supporting Information.
(10) FS is the change in myocyte length divided by the resting
length of the cell. ΔFS is the percent change in FS after
treatment relative to FS at baseline, which is defined as 100%.
(11) Direct vasodilatation effects were measured by the ability of
compounds to relax a potassium chloride (KCl)- or a phenyl-
ephrine-induced contraction of a rat aortic ring. The vasodi-
latation effect of 8 in rat aortic rings was shown to be inde-
pendent of endothelium, protein kinase G, and protein kinase
A pathways for vascular relaxation. However, a dose-related
inhibition of the vasodilatation effect of 8 was observed when
the aortic ring preparations were pretreated with the KATP
channel inhibitor glybenclamide. Therefore, the vasodilata-
tion effect of 8 was determined to be due to its ability to
activate KATP channels.
Figure 2. Concentration-dependent increase in placebo-corrected
absolute fraction shortening in rats by CK-1317138 and omecamtiv
mecarbil. N=5-6 per data point. Points within the dashed lines were
significantly different from placebo (p < 0. 05).
SUPPORTING INFORMATION AVAILABLE Experimental
procedures and analytical data. This material is available free of
AUTHOR INFORMATION
Corresponding Author: *To whom correspondence should be
addressed. Tel: 650-624-3003. Fax: 650-614-3010. E-mail: bmorgan@
cytokinetics.com.
AssayCatalog.aspx?path=128&leaf=128.
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structure-activity relationships of trisubstituted phenyl urea
derivatives as neuropeptide Y5 receptor antagonists. J. Med.
Chem. 2001, 44, 2344–2356.
ACKNOWLEDGMENT We thank all members of the much
larger Cardiovascular Program Team at Cytokinetics for their
contributions.
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Preparation of N,N0-Unsymmetrically Disubstituted Ureas
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(17) FS, measured by echocardiographic imaging, is an assess-
ment of cardiac contractility. FS = (end diastolic diameter -
end systolic diameter)/end diastolic diameter ꢀ 100 as
determined by standard analysis of the M-mode images.
(18) Benet, L. Z.; Hoener, B. Changes in plasma protein binding
have little clinical relevance. Clin. Pharm. Ther. 2002, 71 (3),
115–121.
(19) Kim, H. S.; Mallic, R.; Hage, D. S. Chromatographic analysis of
carbamazepine binding to human serum albumin. II. Com-
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references therein.
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A powerful tool for studying serum protein binding. J. Chro-
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ABBREVIATIONS ATP, adenosine triphosphate; DIPEA, di-
isopropylethylamine; PK, pharmacokinetic; SAR, structure-
activity relationship.
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2010 American Chemical Society
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DOI: 10.1021/ml100138q ACS Med. Chem. Lett. 2010, 1, 472–477
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