Journal of Medicinal Chemistry
Article
98.5%, RT = 5.98 min. 1H NMR (400 MHz,DMSO-d6): δ = 11.06 (s,
1H), 8.80 (s, 1H), 7.88 (d, 1H, J = 9.0 Hz), 7.43 (d, 1H, J = 6.6 Hz),
7.08 (d, 1H, J = 9.0 Hz), 6.35 (s, 1H), 5.61 (m, 1H), 5.42 (s, 1H), 5.10
(t, 1H, J = 9.8 Hz), 4.50−4.40(m, 4H), 4.35 (m, 1H), 3.71 (t, 1H, J =
6.9 Hz), 2.92−2.87 (m, 1H), 2.68- 2.57 (m, 2H), 2.52−2.28 (m, 10H),
1.76−0.90 (m, 20H), 0.80 (t, 3H, J = 7.4 Hz).
Step 2. 2-Methyl-2H-1,2,3-triazole-4-carboxylic acid methyl ester
(263 mg, 1.86 mmol) was charged in a round-bottom flask. Then a
mixture containing THF (15.0 mL), 1 M solution NaOH (9.30 mL,
9.30 mmol, 5.0 equiv), and MeOH (5.00 mL) was added. The solution
was stirred at room temperature. After 4 h, 1 N HCl was added (10.0
mL) and the solvent was evaporated. EtOAc was added, and layers
were separated. Solvent was evaporated. 2-Methyl-2H-1,2,3-triazole 4-
carboxylic acid was obtained as a white solid (215 mg, 91%). 1H NMR
(400 MHz, DMSO-d6): δ 8.17 (s, 1H), 4.22 (s, 3H). FIA MS
(electrospray): 128.0 (M + H)+.
Compound 9. Boc protected macrocyclic amine 1 (100 mg, 0.136
mmol) was charged in a vial with a 4 N solution of HCl in dioxane
(3.0 mL). The solution was stirred at room temperature for 1 h, after
which the solution was evaporated to dryness to provide the amine
hydrochloride which was used as such. 1H-Pyrazole-3-carboxylic acid
(18.3 mg, 0.163 mmol, 1.20 equiv) was dissolved in DMF (2.0 mL).
Triethylamine (75.9 μL, 0.545 mmol, 4.0 equiv) and HATU (62.1 mg,
0.163 mmol, 1.20 equiv) were added, and the mixture was stirred for
15 min. The Boc deprotected macrocyclic amine hydrochloride was
dissolved in DMF (2.0 mL) and added to the acid solution. The
mixture was stirred at room temperature overnight. The resulting
solution was filtered through a Millex filter and purified by preparative
HPLC (X-Bridge column, 10 nM ammonium bicarbonate, pH 10,
MeOH). The pure fractions were combined, concentrated, frozen, and
lyophilized to provide compound 9 as a white amorphous solid (107
mg, 99%). FIA MS (electrospray): 790.1 (M − H)−, 792.2 (M + H)+ .
Reverse phase HPLC homogeneity at 220 nm (0.1% TFA, CH3CN/
Compound 20. Boc protected amine 24 (90 mg, 0.12 mmol) was
dissolved in DCM (4.0 mL), and a commercial solution of 4 N HCl in
dioxane (4.0 mL, 4.0 mmol, 33 equiv) was added. This mixture was
stirred at room temperature for 60 min and then concentrated under
reduced pressure to afford a residue corresponding to the unprotected
amine. The crude intermediate was dissolved in DMF (1.5 mL) along
with TEA (81 μL, 0.58 mmol, 5.0 equiv), 2-methyl-2H-1,2,3-triazole 4-
carboxylic acid (18 mg, 0.14 mmol, 1.2 equiv), and TBTU (53 mg,
0.14 mmol, 1.2 equiv). The solution was stirred at room temperature
for 16 h. The crude mixture was filtered with a Millex filter and
purified directly by preparative HPLC. The appropriate fractions were
combined, frozen, and lyophilized to give compound 20 as a white
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lyophilized solid (37 mg, 39%). H NMR (400 MHz,DMSO-d6): δ
1
10.80 (s, 1H), 8.92 (s, 1H), 8.07 (s, 1H), 8.35 (d, 1H, J = 6.7 Hz), 7.85
(d, 1H, J = 8.8 Hz), 7.10 (d, 1H, J = 8.8 Hz), 6.35 (s, 1H), 5.67−5.59
(m 1H), 5.53−5.43 (m, 2H), 5.10−5.03 (m, 1H), 4.62−4.52 (m, 2H),
4.45−4.39 (m, 1H), 4.20 (s, 3H), 4.06−3.98 (m, 1H), 3.89 (s, 3H),
2.69−2.66 (m, 1H), 2.44 (s, 3H), 2.40−2.31 (m, 2H), 2.06−1.94 (m,
1H), 1.89−1.76 (m, 1H), 1.62−1.49 (m, 2H), 1.47−1.35 (m, 14H),
1.34−1.20 (m, 5H), 0.94−0.83 (m, 2H). FIA MS (electrospray): 821.5
(M + H)+; 819.3 (M − H)−.
H2O): 100%, tR = 5.247 min. H NMR (400 MHz, DMSO-d6): δ =
11.05 (s, 1H), 8.79 (s, 1H), 8.04 (bs, 1H), 7.82 (d, 1H, J = 9 Hz), 7.74
(d, 1H, J = 2.0 Hz), 7.07 (d, 1H, J = 9.0 Hz), 6.71 (s, 1H), 6.37 (s,
1H), 5.61 (dd, 1H, J = 8.3, 18.4 Hz), 5.49 (p, 1H, J = 5.8 Hz), 5.45 (bs,
1H), 5.14 (dd, 2H, J = 9, 10.2 Hz), 4.62−4.54 (m, 2H), 4.34 (dd, 1H, J
= 7.0, 9.8 Hz), 4.02 (dd, 1H, J = 3.5, 11.3 Hz), 3.87 (s, 3H), 2.94−2.88
(m, 1H), 2.62- 2.56 (m, 2H), 2.43 (s, 3H), 2.38−2.31 (m, 2H), 2.01−
1.92 (m, 1H), 1.80−1.75 (m, 1H), 1.58−1.52 (m, 3H), 1.44−1.36 (m,
10H), 1.31−1.20 (m, 2H), 1.11−0.97 (m, 4H).
Compound 19. Intermediate 24 (2.55 g, 3.14 mmol) was charged
in a vial. Then a 4 N solution of HCl in dioxane (40.0 mL, 160 mmol)
was added. The solution was stirred at room temperature for 2 h, after
which a precipitate had formed. The solution was evaporated to
dryness to give the amine hydrochloride. 1-Methyl-1H-pyrazole 4-
carboxylic acid (475 mg, 3.77 mmol, 1.20 equiv) was dissolved in
DMF (10 mL). Then triethylamine (2.19 mL, 15.7 mmol, 5.0 equiv)
was added followed by TBTU (1.43 g, 3.77 mmol, 1.20 equiv). The
solution was stirred for 15 min, after which the amine hydrochloride
was added in DMF (10 mL) and the solution was stirred at room
temperature for 16 h. Work up was the following: Water (25 mL) was
added. The organic layer was extracted with EtOAc (3 × 50 mL) and
dried over MgSO4. The solvent was evaporated. The residue was
purified on CombiFlash (25−70% EtOAc/hexane). The pure fractions
were combined, concentrated, frozen, and lyophilized to provide
compound 19 (1.90 g, 74%). FIA MS (electrospray): 820 (M + H)+.
Reverse phase HPLC homogeneity at 220 nm (0.1% TFA, CH3CN/
H2O): 99%. 1H NMR (400 MHz,DMSO-d6): δ = 10.83 (s, 1H), 8.93
(s, 1H), 7.83 (d, 1H, J = 8.8 Hz), 7.79 (d, 1H, J = 7.0 Hz), 7.77 (d, 1H,
J = 2.1 Hz), 7.08 (d, 1H, J = 9.0 Hz), 6.60 (d, 1H, J = 2.3 Hz), 6.37 (s,
1H), 5.66−5.58 (m, 1H), 5.62 (S, 1H J = 6.2 Hz), 5.47−5.44 (m, 1H),
5.07 (dd, 1H, J = 9.5, 9.2 Hz), 4.64−4.54 (m, 1H) 4.52 (d, 1H, J =
11.6 Hz), 4.40 (dd, 1H, J = 9.7, 7.1 Hz), 4.02 (dd, 1H, J = 11.8, 3.5
Hz), 3.89 (s, 3H), 3.88 (s, 3H), 2.66−2.57 (m, 1H), 2.51 (s, 3H), 2.43
(s, 3H), 2.38−2.29 (m, 2H), 2.02−1.90 (m, 1H), 1.88−1.77 (m, 1H),
1.58 (dd, 1H, J = 8.2, 5.1 Hz), 1.52 (dd, 1H, J = 9.3, 5.2 Hz), 1.45−
1.35 (m, 12H), 1.34−1.20 (m, 4H), 0.93−0.84 (m, 2H).
ASSOCIATED CONTENT
* Supporting Information
■
S
Detailed synthesis of macrocycle 21, quinolines 22b,c,
carbamates 2−6, and amides 7−20. This material is available
AUTHOR INFORMATION
Corresponding Author
■
Notes
The authors declare no competing financial interest.
ACKNOWLEDGMENTS
■
The authors express their gratitude to the biology, analytical,
and structural research department members who contributed
to the generation of the data. The authors also thank Drs.
Richard Bethell and Michael Cordingley for their guidance and
support during this work.
ABBREVIATIONS USED
■
ADME, administration, distribution, metabolism, and excretion;
AUC, area under the curve; CYP, cytochrome P 450; DCM,
dichloromethane; DAA, direct acting antiviral; DIPEA,
diisopropylethylamine; DMAP, dimethylaminopyridine;
DMSO, dimethylsulfoxide; CDCl3, deuterated chloroform;
EtOAc, ethyl acetate; HCV, hepatitis C virus; HPLC, high
pressure liquid chromatography; PEG, polyethylene glycol; PK,
pharmacokinetic; LCIQ, liver-corrected inhibitory quotient;
LiHMDS, lithium bis(trimethylsilyl)amide; MES, 2-(N-
morpholino)ethanesulfonic acid; SVR, sustained virological
response; TBTU, O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethy-
luronium tetrafluoroborate; TCEP, tris(2-carboxyethyl)-
Synthesis of 2-Methyl-2H-1,2,3-triazole-4-carboxylic Acid.
Step 1. Methyl cyanoformate (1.00 g, 11.7 mmol) was charged in a
flask and dissolved in THF (40.0 mL). Then a 0.6 M diazomethane
solution in Et2O (58.8 mL, 35.3 mmol, 3.00 equiv) was added. This
solution was stirred at room temperature for 16 h. Water (40 mL) and
EtOAc (40 mL) were added, and then the layers were separated. The
solvent was evaporated and purification was performed on Combiflash
(20−100% hexane) to provide the 3,4-regioisomer (654 mg, 39%) and
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the desired 2,4-regioisomer (434 mg, 26%) as a clear yellow oil. H
NMR (400 MHz,CDCl3): δ 8.05 (s, 1H), 4.28 (s, 3H) 3.96 (s, 3H).
FIA MS (electrospray): 142.2 (M + H)+.
F
dx.doi.org/10.1021/jm400121t | J. Med. Chem. XXXX, XXX, XXX−XXX