7196 Journal of Medicinal Chemistry, 2006, Vol. 49, No. 24
Marsault et al.
mycin and Cisapride on gastric Emptying in Dogs. J. Pharm. Exp.
Ther. 2000, 293, 1106-1111. (b) Russo, A.; Stevens, J. E.; Giles,
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338.
HPLC-CLND detector, and found to be 13 mg (21% overall yield
based on the effective loading of Nva).
RP-HPLC Purification. Crude macrocycle 16i was purified on
a Waters FractionLynx MS-triggered reverse phase preparative
system, using a water:methanol gradient containing 0.1% TFA as
the eluent. Macrocycle 16i was obtained as a colorless film (5.2
mg as quantified by HPLC-CLND). HPLC purity: 98.8% (UV
detector), 100% (ELSD detector), 100% (CLND detector).
HRMS: C31H44N4O5, calc: 552.3311, found: 552.3321 ( 0.0016.
1H NMR (300 MHz, CD3OD) δ (ppm) 8.77 (d, 1H, 9 Hz); 7.75 (d,
1H, 3 Hz); 7.21-7.05 (m, 4H); 6.87-6.73 (m, 4H); 4.46-4.38
(m, 1H); 4.15-4.00 (m, 3H); 3.79 (d, 1H, 12 Hz); 3.74 (s, 3H);
3.29-3.04 (m, 5H); 2.71 (dt, 1H, 12 Hz; 6 Hz); 2.63-2.53 (m,
1H); 2.20 (dt, 1H, 12 Hz; 3 Hz); 2.13-1.98 (m, 2H); 1.66-1.33
(m, 5H); 1.12 (s, 3H, 6 Hz); 1.02 (d, 3H, 6 Hz); 0.96 (t, 3H, 6 Hz).
13C NMR (75.5 MHz, CD3OD) δ (ppm) 174.80; 174.21; 160.44;
127.31; 132.47; 131.46; 131.32; 128.43; 122.11; 114.89; 111.41;
65.88; 63.69; 61.48; 55.59; 54.20; 48.10; 42.40; 38.48; 34.31; 31.32;
30.09; 29.08; 20.57; 20.26; 19.52; 13.67.
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Typically, compounds were obtained as TFA salts. They were
dissolved at 10 mM concentration in DMSO in a 96-well plates
and stored at -60 °C until tested. For characterization purposes,
selected macrocycles underwent a salt exchange step using ion-
exchange resin to generate the corresponding hydrochloride salts.
Acknowledgment. We thank Mr. Patrick Bhe´rer, Mrs.
Manon Champagne, Mrs. Maude Gauthier, Mrs. Annie Letendre
as well as Dr. Axel Mathieu and Dr. Rene´ Gagnon for analytical
support of this work. Mr. Gaston Boulay (U. Sherbrooke, QC)
is acknowledged for performing HRMS analyses.
Supporting Information Available: Experimental procedures
for biological assays; the synthesis of macrocycles (method B),
including procedures for the synthesis of tether and linker building
blocks; characterization data for building blocks (LC/MS, 1H, NMR,
1
13C NMR) and macrocycles (HPLC/MS, HRMS, H NMR, 13C
NMR), including spectra and chromatograms. This material is
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