10.1002/cmdc.201800366
ChemMedChem
FULL PAPER
126.8, 46.7, 42.1, 37.0, 36.7, 25.9, 24.9; m/z (ES+) 393 MH+
(30%); HRMS C20H37N6O2 (MH+) calculated 393.2978, found
393.2976.
8.0 Hz), 7.34 (4H, d, J = 8.0 Hz); dC (75.5 MHz, D2O, amide
rotamers) 174.5, 139.9, 133.5, 130.5, 129.4, 126.2, 46.6, 42.0,
37.0, 36.7, 25.9, 24.9; m/z (ES+) 495 MH+ (20%); HRMS
C28H43N6O2 (MH+) calculated 495.3447, found 495.3445.
(8a): dH (300 MHz, D2O, amide rotamers) 3.11 (4H, approx. t,
J = 7.5 Hz), 3.31 (4H, t, J = 6.0 Hz), 3.63 (4H, approx. t, J = 7.5
Hz), 3.84 (4H, t, J = 6.0 Hz), 7.56 (1H, s), 7.61-7.65 (3H, m);
dC (75.5 MHz, D2O, amide rotamers) 174.1, 134.9, 130.2,
128.3, 124.2, 46.7, 43.5, 37.6, 36.7; m/z (ES+) 337 MH+ (10%);
HRMS C16H29N6O2 (MH+) calculated 337.2352, found
337.2351.
(3c): dH (300 MHz, D2O, amide rotamers) 1.35-2.03 (12H, m),
2.69-2.80 (4H, m), 3.00-3.05 (4H, m), 3.26-3.36 (4H, m), 3.47-
3.59 (4H, m), 6.76 (2H, s), 7.21-7.26 (4H, m), 7.33 (4H, d, J =
8.0 Hz); dC (75.5 MHz, D2O, amide rotamers) 174.4, 174.0,
139.0, 133.9, 133.8, 130.7, 129.4, 129.3, 126.3, 126.2, 49.9,
48.9, 46.6, 44.6, 42.0, 39.2, 38.9, 37.0, 36.8, 25.9, 24.9, 24.3,
23.9; m/z (ES+) 523 MH+ (15%); HRMS C30H47N6O2 (MH+)
calculated 523.3760, found 523.3752.
(8b): dH (300 MHz, D2O, amide rotamers) 1.87 (4H, approx.
quint., J = 8.0 Hz), 2.04 (4H, quint., J = 7.5 Hz), 2.75 (4H,
approx. t, J = 8.0 Hz), 3.06 (4H, approx. t, J = 7.5 Hz), 3.34
(4H, approx. t, J = 7.5 Hz), 3.60 (4H, t, J = 7.0 Hz), 7.42 (1H,
s), 7.51-7.64 (3H, m); dC (75.5 MHz, D2O, amide rotamers)
173.4, 135.7, 129.9, 127.6, 123.7, 46.7, 42.3, 37.1, 36.7, 25.9,
(5): dH (300 MHz, D2O, amide rotamers) 1.91 (4H, quint., J =
7.0 Hz), 2.05 (4H, quint., J = 7.0 Hz), 2.77 (4H, t, J = 7.5 Hz),
3.07 (4H, t, J = 7.0 Hz), 3.40 (4H, t, J = 7.5 Hz), 3.62 (4H, t, J
= 7.0 Hz), 7.32 (2H, s), 7.42 (4H, d, J = 8.0 Hz), 7.70 (4H, d, J
= 8.0 Hz); dC (75.5 MHz, D2O, amide rotamers) 174.5, 138.9,
134.0, 129.2, 127.1, 126.8, 46.7, 42.1, 37.1, 36.7, 25.9, 24.0;
m/z (ES+) 495 MH+ (5%); HRMS C28H43N6O2 (MH+) calculated
495.3447, found 495.3443.
25.0; m/z (ES+) 393 MH+ (20%), 197 MH2 (70%); HRMS
2+
C20H37N6O2 (MH+) calculated 393.2978, found 393.2968.
(8c): dH (300 MHz, D2O, amide rotamers) 1.37-2.08 (12H, m),
2.72-2.82 (4H, m), 3.03-3.08 (4H, m), 3.25-3.36 (4H, m), 3.50-
3.62 (4H, m), 7.39 (1H, s), 7.49-7.60 (3H, m); dC (75.5 MHz,
D2O, amide rotamers) 173.3, 172.9, 135.8, 129.8, 127.6,
127.5, 123.6, 49.0, 46.7, 44.8, 42.2, 39.2, 38.9, 37.0, 36.7,
25.9, 25.0, 24.3, 23.9; m/z (ES+) 421 MH+ (20%); HRMS
C22H41N6O2 (MH+) calculated 421.3291, found 421.3289.
(7a): dH (300 MHz, D2O, amide rotamers) 3.22 (4H, t, J = 7.0
Hz), 3.43 (4H, t, J = 6.0 Hz), 3.81 (4H, t, J = 7.0 Hz), 3.96 (4H,
t, J = 6.0 Hz), 7.57 (2H, d, J = 7.5 Hz), 7.71 (2H, t, J = 8.0 Hz),
7.83 (2H, s), 7.91 (2H, d, J = 8.0 Hz); dC (75.5 MHz, D2O,
amide rotamers) 175.1, 140.5, 134.7, 129.9, 129.3, 125.6,
124.9, 46.8, 43.5, 37.8, 36.8; m/z (ES+) 413 MH+ (20%); HRMS
C22H33N6O2 (MH+) calculated 413.2665, found 413.2660.
4.1.7 Scheme 7 (BIT 180, 181, 241, 242, 228, 283):
(7b): dH (300 MHz, D2O, amide rotamers) 1.92 (4H, approx.
quint., J = 7.5 Hz), 2.08 (4H, approx. quint. J = 7.0 Hz), 2.77
(4H, t, J = 7.5 Hz), 3.10 (4H, approx. t, J = 7.5 Hz), 3.43 (4H, t,
J = 7.5 Hz), 3.43 (4H, t, J = 7.0 Hz), 7.44 (2H, d, J = 7.5 Hz),
7.63 (2H, t, J = 7.5 Hz), 7.68 (2H, s), 7.82 (2H, d, J = 7.5 Hz);
dC (75.5 MHz, D2O, amide rotamers) 174.4, 140.5, 135.6,
129.8, 128.9, 125.5, 124.6, 46.7, 42.1, 37.1, 36.7, 25.9, 25.0;
m/z (ES+) 469 MH+ (80%); HRMS C26H41N6O2 (MH+) calculated
469.3291, found 469.3300.
General : cis-Stilbene-4,4’-dicarboxylic acid (1) and its trans
isomer (2) are commercially available (Lancaster). Biphenyl-
3,3’-dicarboxylic acid (6) was prepared by literature methods
[35]. Di-protected triamines were prepared by known methods
referenced in scheme 6.
Compound
(3a):
BOP
(Benzotriazol-1-yloxy-
tris(dimethylamino)-phosphonium hexafluorophosphate) (52
mg, 0.12 mmol) was added to a solution of cis-stilbene-4,4’-
dicarboxylic acid (15 mg, 0.056 mmol) in dry DMF (1 ml) and
the mixture stirred for 5 min at room temperature under N2. [2-
(2-tert-Butoxycarbonylaminoethylamino)ethyl]carbamic acid
tert-butyl ester (36 mg, 0.12 mmol) and Et3N (19 µl, 0.14 mmol)
were added and stirring continued overnight. Water (30 ml)
was added and the mixture extracted with EtOAc (3 x 20 ml).
The organic extracts were washed with sat. NaCl (aq., 30 ml),
dried (Na2SO4) and the solvent removed. The derivative (2a)
was purified by flash chromatography on silica gel (2.5%
MeOH / EtOAc) to give 31 mg, 66%. This material (28 mg) was
suspended in DCM (8 ml) and TFA (1.2 ml) added dropwise.
The mixture was stirred for 6 h at room temperature, the
solvent was removed and the residue washed with several
portions of Et2O to give (3a) as a hygroscopic solid (26 mg,
87% if x = 4).
4.1.8 Scheme 8 (BIT 160):
1,4,5,8-Tetramethylnaphthalene (3) was prepared by a known
method [36] and isomerized to 1,3,5,7–tetramethylnapthalene
(4) as described by Oku and Yuzen[37]. This material was
brominated and converted to the final product (7) using similar
procedures to those described in scheme 1 for related
compounds prepared from m-xylene.
(7): dH (300 MHz, D2O) 3.01-3.07 (8H, m), 3.62 (8H, t, J = 5.0
Hz), 4.65 (4H, s), 5.95 (4H, s), 7.54 (2H, s), 7.97 (2H, s); dC
(75.5 MHz, CD3OD) 136.5, 135.6, 132.8, 128.2, 124.6, 74.4,
72.6, 67.2, 67.0, 40.7; m/z (ES+) 421 MH+ (15%); HRMS
C22H37N4O4 (MH+) calculated 421.2815, found 421.2798.
(3a): dH (300 MHz, D2O, amide rotamers) 3.10 (4H, t, J = 7.0
Hz), 3.28 (4H, approx. t, J = 6.0 Hz), 3.66 (4H, t, J = 7.0 Hz),
3.80 (4H, approx. t, J = 6.0 Hz), 6.76 (2H, s), 7.30 (4H, d, J =
8.0 Hz), 7.36 (4H, d, J = 8.0 Hz); dC (75.5 MHz, D2O, amide
rotamers) 175.2, 139.4, 132.6, 130.7, 129.5, 126.5, 46.7, 43.4,
37.7, 36.8; m/z (ES+) 439 MH+ (20%); HRMS C24H35N6O2
(MH+) calculated 439.2821, found 439.2823.
4.2 Isolation of SR vesicles: Cardiac SR was prepared from
sheep heart [15]Vesicles were frozen and stored either in liquid
N2 or at -70°C. Skeletal SR was isolated from the back and leg
muscles of New Zealand White rabbits, and heavy SR was
collected from the 35-45% (wt/vol) interface of a discontinuous
sucrose gradient, centrifuged and resuspended [38].
Compounds (3b), (3c), (5), (7a), (7b): Prepared by the
applying the same procedure used to make (3a) to the
appropriate diacid and triamine starting materials.
4.3 Ca2+ release. Extravesicular Ca2+ was monitored at
710 nm with the Ca2+ indicator, antipyrylazo III, using a Cary 3
Spectrophotometer[6]. Identical release experiments were
performed at 790 nm, to detect Ca2+-independent changes in
optical density (OD) which would alter the Ca2+ release
measurement. The cuvette solution was stirred continuously
(3b): dH (300 MHz, D2O, amide rotamers) 1.86 (4H, approx.
quint., J = 7.5 Hz), 2.00 (4H, quint. J = 7.0 Hz), 2.72 (4H,
approx. t, J = 7.5 Hz), 3.02 (4H, t, J = 7.5 Hz), 3.34 (4H, t, J =
7.5) Hz), 3.57 (4H, t, J = 7.0 Hz), 6.76 (2H, s), 7.24 (4H, d, J =
20
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