Journal of Medicinal Chemistry
Article
31 (58.5 mg, 0.289 mmol) according to the method described for the
synthesis of 5. H NMR (400 MHz, CD3OD): δ 7.35−7.32 (m, 2H),
pyran-3,4,5-triol (40).53 To a stirred solution of 27 (100 mg, 0.261
mmol) and 30 (58.9 mg, 0.287 mmol) in N,N-dimethylformamide (1.5
mL) was added cesium carbonate (258 mg, 0.79 mmol) followed by
addition of tetrabutylammonium iodide (9.7 mg, 0.026 mmol). After
stirring for 12 h at 40 °C under argon, the mixture was quenched by
addition of ice water (10 mL). The mixture was extracted with ethyl
acetate (2 × 20 mL). The combined organic layers were washed with
water (20 mL) and brine (20 mL), dried over anhydrous sodium sulfate,
and concentrated, and the residue was purified by preparative TLC to
give product 40 (81.6 mg, 75% yield, HPLC purity (condition I):
99.9%) as a white solid. 1H NMR (400 MHz, CD3OD): δ 7.35−7.32 (m,
2H), 7.27 (dd, J = 2.0, 8.4 Hz, 1H), 7.09 (d, J = 8.4 Hz, 2H), 6.79 (d, J =
8.4 Hz, 2H), 4.09 (d, J = 9.6 Hz, 1H), 3.87 (d, J = 12.4 Hz, 1H), 3.69 (dd,
J = 4.8, 11.6 Hz, 1H), 3.47−3.43 (m, 1H), 3.42−3.35 (m, 2H), 3.29−
3.26 (m, 1H). 13C NMR (100 MHz, CD3OD): δ 158.9, 140.0, 139.9,
134.4, 132.8, 131.9, 130.8, 130.1, 128.2, 115.4, 82.2, 79.8, 76.4, 71.9,
1
7.27 (dd, J = 1.6, 8.0 Hz, 1H), 7.09 (d, J = 8.8 Hz, 2H), 6.79 (d, J = 8.8
Hz, 2H), 4.09 (d, J = 9.2 Hz, 1H), 4.05 (d, J = 15.2 Hz, 1H), 3.99 (d, J =
15.2 Hz, 1H), 3.87 (d, J = 12.0 Hz, 1H), 3.69 (dd, J = 5.2, 12.0 Hz, 1H),
3.48−3.44 (m, 1H), 3.43−3.37 (m, 2H), 3.30−3.26 (m, 1H), 1.33 (s,
3H). 13C NMR (100 MHz, CD3OD): δ 158.8, 140.0, 139.9, 134.4,
132.9, 131.9, 130.8, 130.1, 128.2, 115.4, 82.9, 82.2, 79.7, 76.4, 71.8, 63.1,
39.2, 14.9. MS (ESI) m/z 411 (M + H+, positive mode), 455 (M +
−
HCO2 , negative mode). HRMS: m/z calcd for C21H22D2ClO6 (M −
H+), 409.1393; found, 409.1406.
(2R,3R,4S,5S,6R)-2-(4-Chloro-3-(4-ethoxybenzyl)phenyl)-2-d-
6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (36). Com-
pound 36 (80.5 mg, 75% yield, HPLC purity (condition I): 99.7%)
was prepared as a white solid from 25 (100 mg, 0.262 mmol) and 29
(57.7 mg, 0.288 mmol) according to the method described for the
1
−
63.1. MS (ESI) m/z 417 (M + H+, positive mode), 461 (M + HCO2 ,
synthesis of 5. H NMR (400 MHz, CD3OD): δ 7.35−7.32 (m, 2H),
negative mode). HRMS: m/z calcd for C21H17D8ClO6 (M − H+),
415.1842; found, 415.1769.
7.27 (d, J = 8.4 Hz, 1H), 7.09 (d, J = 8.4 Hz, 2H), 6.79 (d, J = 8.4 Hz,
2H), 4.07−4.01 (m, 2H), 3.98 (q, J = 7.2 Hz, 2H), 3.87 (d, J = 12.0 Hz,
1H), 3.69 (dd, J = 4.8, 12.0 Hz, 1H), 3.48−3.43 (m, 1H), 3.42−3.35 (m,
2H), 3.29−3.26 (m, 1H), 1.35 (t, J = 7.2 Hz, 3H). 13C NMR (100 MHz,
CD3OD): δ 158.8, 140.0, 139.9, 134.4, 132.9, 131.9, 130.8, 130.1, 128.2,
115.4, 82.2, 79.7, 76.4, 71.8, 64.4, 63.1, 39.2, 15.2. MS (ESI) m/z 410 (M
(2S,3R,4R,5S,6R)-2-(4-Chloro-3-(4-(methoxy-d3)benzyl)-
phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol
(41). Compound 41 (75.3 mg, 72% yield, HPLC purity (condition I):
99.1%) was prepared as a white solid from 11 (100 mg, 0.263 mmol) and
33 (54.8 mg, 0.289 mmol) according to the method described for the
synthesis of 5. 1H NMR (400 MHz, CD3OD): δ 7.34 (d, J = 8.0 Hz, 1H),
7.32 (d, J = 2.0 Hz, 1H), 7.27 (dd, J = 2.0, 8.0 Hz, 1H), 7.10 (d, J = 8.8
Hz, 2H), 6.81 (d, J = 8.8 Hz, 2H), 4.10−3.98 (m, 3H), 3.87 (d, J = 11.6
Hz, 1H), 3.69 (dd, J = 5.2, 11.6 Hz, 1H), 3.48−3.43 (m, 1H), 3.42−3.35
(m, 2H), 3.30−3.27 (m, 1H). 13C NMR (100 MHz, CD3OD): δ 159.6,
140.0, 139.9, 134.4, 132.9, 131.9, 130.8, 130.1, 128.2, 114.8, 82.9, 82.2,
−
+ H+, positive mode), 454 (M + HCO2 , negative mode). HRMS: m/z
calcd for C21H24DCl2O6 (M + Cl−), 444.1096; found, 444.1101.
(2S,3R,4R,5S,6R)-2-(4-Chloro-3-((4-ethoxyphenyl)methyl-d2)-
phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol
(37). Compound 37 (75.1 mg, 70% yield, HPLC purity (condition I):
99.0%) was prepared as a white solid from 26 (100 mg, 0.261 mmol) and
29 (57.5 mg, 0.287 mmol) according to the method described for the
1
synthesis of 5. H NMR (400 MHz, CD3OD): δ 7.35−7.32 (m, 2H),
+
79.7, 76.5, 71.8, 63.1, 39.2. MS (ESI) m/z 415 (M + NH4 , positive
7.27 (dd, J = 2.4, 8.4 Hz, 1H), 7.09 (d, J = 8.8 Hz, 2H), 6.79 (d, J = 8.8
Hz, 2H), 4.09 (d, J = 9.6 Hz, 1H), 3.99 (q, J = 7.2 Hz, 2H), 3.87 (d, J =
11.6 Hz, 1H), 3.69 (dd, J = 5.2, 11.6 Hz, 1H), 3.48−3.43 (m, 1H), 3.42−
3.36 (m, 2H), 3.29−3.26 (m, 1H), 1.35 (t, J = 7.2 Hz, 3H). 13C NMR
(100 MHz, CD3OD): δ 158.8, 140.0, 139.9, 134.4, 132.8, 131.9, 130.8,
130.1, 128.2, 115.4, 82.9, 82.2, 79.7, 76.4, 71.8, 64.4, 63.1, 15.2. MS
−
mode), 442 (M + HCO2 , negative mode). HRMS: m/z calcd for
C20H20D3ClNaO6 (M + Na+), 420.1264; found, 420.1264.
(2S,3R,4R,5S,6R)-2-(4-Chloro-3-((4-(methoxy-d3)phenyl)-
methyl-d2)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-
3,4,5-triol (42). Compound 42 (73.0 mg, 70% yield, HPLC purity
(condition I): 99.6%) was prepared as a white solid from 26 (100 mg,
0.261 mmol) and 33 (54.3 mg, 0.287 mmol) according to the method
described for the synthesis of 5. 1H NMR (400 MHz, CD3OD): δ 7.35−
7.32 (m, 2H), 7.27 (dd, J = 2.0, 8.4 Hz, 1H), 7.10 (d, J = 8.8 Hz, 2H),
6.80 (d, J = 8.8 Hz, 2H), 4.09 (d, J = 9.6 Hz, 1H), 3.87 (d, J = 12.0 Hz,
1H), 3.69 (dd, J = 5.2, 12.0 Hz, 1H), 3.48−3.43 (m, 1H), 3.42−3.36 (m,
2H), 3.30−3.27 (m, 1H). 13C NMR (100 MHz, CD3OD): δ 159.6,
140.0, 139.9, 134.4, 132.9, 131.9, 130.8, 130.1, 128.2, 114.8, 82.9, 82.2,
79.7, 76.4, 71.8, 63.1. MS (ESI) m/z 422 (M + Na+, positive mode), 444
−
(ESI) m/z 411 (M + H+, positive mode), 455 (M + HCO2 , negative
mode). HRMS: m/z calcd for C21H23D2Cl2O6 (M + Cl−), 445.1159;
found, 445.1168.
(2S,3R,4R,5S,6R)-2-(4-Chloro-3-((4-(ethoxy-d5)phenyl)-
methyl-d2)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-
3,4,5-triol (38). Compound 38 (73.8 mg, 68% yield, HPLC purity
(condition I): 98.8%) was prepared as a white solid from 26 (100 mg,
0.261 mmol) and 30 (58.9 mg, 0.287 mmol) according to the method
described for the synthesis of 5. 1H NMR (400 MHz, CD3OD): δ 7.35−
7.32 (m, 2H), 7.27 (dd, J = 2.0, 8.0 Hz, 1H), 7.09 (d, J = 8.4 Hz, 2H),
6.79 (d, J = 8.4 Hz, 2H), 3.87 (d, J = 12.0 Hz, 1H), 3.69 (dd, J = 5.2, 12.0
Hz, 1H), 3.47−3.43 (m, 1H), 3.42−3.35 (m, 2H), 3.29−3.26 (m, 1H).
13C NMR (100 MHz, CD3OD): δ 158.9, 140.0, 139.9, 134.4, 132.8,
−
(M + HCO2 , negative mode). HRMS: m/z calcd for C20H18D5ClNaO6
(M + Na+), 422.1389; found, 422.1382.
(2S,3R,4R,5S,6R)-2-(4-Chloro-3-((4-(methoxy-d3)phenyl)-
methyl-d2)phenyl)-2-d-6-(hydroxymethyl)tetrahydro-2H-
pyran-3,4,5-triol (43). Compound 43 (75.3 mg, 72% yield, HPLC
purity (condition I): 99.8%) was prepared as a white solid from 27 (100
mg, 0.261 mmol) and 33 (54.3 mg, 0.287 mmol) according to the
method described for the synthesis of 5. 1H NMR (400 MHz, CD3OD):
δ 7.35−7.32 (m, 2H), 7.27 (dd, J = 2.0, 8.0 Hz, 1H), 7.11 (d, J = 8.4 Hz,
2H), 6.80 (d, J = 8.4 Hz, 2H), 3.87 (d, J = 11.6 Hz, 1H), 3.69 (dd, J = 5.2,
11.6 Hz, 1H), 3.48−3.43 (m, 1H), 3.42−3.35 (m, 2H), 3.30−3.27 (m,
1H). 13C NMR (100 MHz, CD3OD): δ 159.6, 140.0, 139.9, 134.5,
132.9, 131.9, 130.8, 130.1, 128.2, 114.8, 82.2, 79.8, 76.4, 71.9, 63.1. MS
131.9, 130.8, 130.1, 128.2, 115.4, 82.9, 82.2, 79.7, 76.5, 71.8, 63.1. MS
−
(ESI) m/z 438 (M + Na+, positive mode), 460 (M + HCO2 , negative
mode). HRMS: m/z calcd for C21H18D7ClNaO6 (M + Na+), 438.1671;
found, 438.1676.
(2S,3R,4R,5S,6R)-2-(4-Chloro-3-((4-ethoxyphenyl)methyl-d2)-
phenyl)-2-d-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol
(39). Compound 39 (78.5 mg, 73% yield, HPLC purity (condition I):
99.1%) was prepared as a white solid from 27 (100 mg, 0.261 mmol) and
29 (57.5 mg, 0.287 mmol) according to the method described for the
+
−
1
(ESI) m/z 418 (M + NH4 , positive mode), 444 (M + HCO2 , negative
mode). HRMS: m/z calcd for C20H16D6ClO6 (M − H+), 399.1488;
found, 399.1475.
synthesis of 5. H NMR (400 MHz, CD3OD): δ 7.35−7.32 (m, 2H),
7.27 (dd, J = 2.0, 8.4 Hz, 1H), 7.09 (d, J = 8.8 Hz, 2H), 6.79 (d, J = 8.8
Hz, 2H), 4.09 (d, J = 9.6 Hz, 1H), 3.97 (q, J = 6.8 Hz, 2H), 3.87 (d, J =
12.0 Hz, 1H), 3.69 (dd, J = 5.2, 12.0 Hz, 1H), 3.48−3.44 (m, 1H), 3.43−
3.35 (m, 2H), 3.30−3.27 (m, 1H), 1.35 (t, J = 6.8 Hz, 3H). 13C NMR
(100 MHz, CD3OD): δ 158.9, 140.0, 139.9, 134.4, 132.9, 131.9, 130.8,
130.1, 128.2, 115.5, 82.2, 79.8, 76.4, 71.9, 64.5, 63.1, 15.2. MS (ESI) m/z
Glucose Uptake Inhibition Assay. A plasmid bearing the human
full-length SGLT1 coding sequence in the pDream 2.1 mammalian
expression vector was purchased from GenScript Corporation. Full-
length human SGLT2 cDNA (GenScript Corporation) was cloned into
the pEAK15 mammalian expression vector. Human SGLT1 expression
plasmid DNA was transfected into COS-7 cells (American Type Culture
Collection) using Lipofectamine 2000 (Invitrogen Corporation).
Transfected cells were evaluated for SGLT1 activity in a methyl-α-D-
[U-14C]glucopyranoside (AMG) uptake assay and cryopreserved until
+
−
429 (M + NH4 , positive mode), 456 (M + HCO2 , negative mode).
HRMS: m/z calcd for C21H21D3ClO6 (M − H+), 410.1456; found,
410.1445.
(2S,3R,4R,5S,6R)-2-(4-Chloro-3-((4-(ethoxy-d5)phenyl)-
methyl-d2)phenyl)-2-d-6-(hydroxymethyl)tetrahydro-2H-
1248
dx.doi.org/10.1021/jm401780b | J. Med. Chem. 2014, 57, 1236−1251