Journal of Medicinal Chemistry
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1.0 equiv) in THF was added triethylamine (162 mg, 1.6 mmol, 3.5
equiv), 4-isopropyl-N-methylaniline (68 mg, 0.457 mmol, 1.0 equiv),
and propylphosphonic anhydride (435 mg, 1.37 mmol, 3.0 equiv) at
0 °C. The reaction mixture was stirred at room temperature for 16 h. It
was quenched with ice water and extracted with ethyl acetate (3 × 50
mL). The organic layer was washed with water (1 × 30 mL), brine
(1 × 30 mL), dried over sodium sulfate, and was evaporated to
dryness. The crude mass was purified by column chromatography
(20% ethyl acetate in hexane) to afford the desired material 29b as a
The reaction was stirred for 30 min. The reaction was judged complete
by LC-MS and quenched with water and extracted with ethyl acetate.
The organic layer was then concentrated and dissolved in DMSO. The
crude product solution in DMSO was purified via prep-HPLC to
provide the desired material (54 mg, 61%). 1H NMR (600 MHz,
DMSO-d6): δ = 1.20 (d, J = 7.02 Hz, 6 H), 1.52−1.98 (m, 4 H), 2.10−
2.35 (m, 1 H), 2.87 (dt, J = 13.59, 6.80 Hz, 1 H), 2.92−3.30 (m, 1 H),
3.58 (m, 6 H), 3.69−4.21 (m, 2 H), 4.48−4.77 (m, 1 H), 5.77−5.98
(m, 1 H), 7.22 (d, J = 8.77 Hz, 2 H), 7.67−7.72 (m, 3 H), 7.86 (d, J =
7.45 Hz, 1 H), 8.06 (d, J = 7.89 Hz, 1 H), 8.23 (br. s., 1 H), 8.32 (s, 1 H),
10.23 (s, 1 H). LC-MS: 100%, m/z =538.3 [M + H]; HRMS [M + H]
for C31H36N7O2, calcd, 538.2925; found, 538.2927.
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brown solid (150 mg, 58%). H NMR (400 MHz, CDCl3): δ = 1.15
(d, 6H). 1.43 (s, 9H), 1.69 (d, 1H), 1.87 (s, 1H), 2.22−2.16 (m, 2H),
2.86−2.80 (m, 2H), 3.49 (s, 3H), 4.12−4.10 (m, 1H), 4.81 (s, 1H),
5.23 (s, 2H), 6.99 (d, 2H), 7.08 (d, 2H), 7.30 (s, 2H), 7.60 (s, 1H),
7.71 (s, 1H), 8.35 (s, 1H); LC-MS: m/z = 568.2 [M + H].
(R,E)-3-(4-Amino-1-(1-(4-hydroxybut-2-enoyl)piperidin-3-yl)-1H-
pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylphenyl)benzamide
(58). By following a procedure similar to that for 62, the free
piperidine prepared for the synthesis of 40 (82 mg, 0.18 mmol, 1.0
Step 2: To a stirred solution of 29b (100 mg, 0.216 mmol,
1.0 equiv) in dioxane (1 mL) was added 4 N HCl in dioxane (7 mL) at
0 °C, and it was stirred at room temperature for 3 h. Then, the excess
dioxane was evaporated. The reaction residue was triturated with
n-pentane. The solid was collected by filtration and dried to afford the
deprotected piperidine. The white solid (130 mg, 91%) was directly
used for the next step as HCl salt. LC-MS: m/z = 470.0 [M + H].
Step 3: To a stirred solution of the deprotected piperidine (130 mg,
0.25 mmol, 1.0 equiv) in DMF (2 mL) was added BOP reagent
(110.6 mg, 0.25 mmol, 1.0 equiv) followed by DIPEA (0.108 mL,
0.625 mmol, 2.5 equiv) and acrylic acid (19.9 mg, 0.276 mmol, 1.1
equiv). The reaction was stirred at room temperature for 10 min.
Then, it was quenched with ice water and filtered. The residue was
dissolved in ethyl acetate, dried over sodium sulfate, and was con-
centrated to dryness. The crude product was purified by prep-HPLC
1
equiv) was converted into 58 (46 mg, 48%). H NMR (600 MHz,
DMSO-d6): δ = 0.65−0.81 (m, 6 H), 1.15 (br. s., 1 H), 1.48 (br. s.,
1 H), 1.69 (d, J = 9.21 Hz, 1 H), 1.83 (d, J = 10.52 Hz, 1 H), 2.42 (dt,
J = 14.03, 7.02 Hz, 1 H), 2.77 (br. s., 1 H), 3.21 (s, 2 H), 3.57−3.78
(m, 2 H), 4.07−4.37 (m, 1 H), 5.95−6.30 (m, 2 H), 6.77 (d, J = 8.33
Hz, 2 H), 7.14−7.30 (m, 3 H), 7.41 (d, J = 7.89 Hz, 1 H), 7.61 (d, J =
7.89 Hz, 1 H), 7.77 (s, 1 H), 7.88 (s, 1 H) 9.77 (s, 1 H). LC-MS:
100%, m/z = 540.3 [M + H]; HRMS [M + H] for C30H34N7O3, calcd,
540.2718; found, 540.2717.
(R,E)-3-(4-Amino-1-(1-(4-(methylamino)but-2-enoyl)piperidin-3-
yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylphenyl)-
benzamide (59). To a solution of the bromocrotonamide obtained in
the synthesis of 61 (100 mg, 0.166 mmol, 1.0 equiv) in THF (10 mL)
was added triethylamine (0.069 mL, 0.498 mmol, 3.0 equiv), methyl
amine (0.830 mmol, 5.0 equiv), and potassium iodide (27.7 mg, 0.166
mmol, 1.0 equiv). The reaction was stirred at room temperature
overnight in a sealed vial. The LC-MS of the reaction mixture showed
completion of the reaction. The reaction was concentrated to dryness
and diluted with DMSO, which was subjected to HPLC purification to
provide the desired material 59 as a solid (2.9 mg, 3%). LC-MS: 99%,
m/z = 553.2 [M + H];
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to afford final compound 54 as a white solid (30 mg, 24%). H NMR
(400 MHz, DMSO-d6, 100 °C): δ = 1.12 (d, 6 H), 1.48−1.28 (m,
1 H), 1.68−1.62 (m, 1 H), 1.99−1.95 (m, 1 H), 2.33−2.15 (m, 1 H),
2.82 (q, 1 H), 3.17 (t, 1 H), 3.40 (s, 3 H), 3.50 (br s, 1 H), 4.11−4.08
(m, 1 H), 4.35 (br s, 1 H), 4.75−4.70 (m, 1 H), 5.63−5.61 (m, 1 H),
6.08−6.04 (m, 2 H), 6.40 (s, 2 H), 6.74−6.67 (m, 1 H), 7.14 (s, 4 H),
7.41−7.38 (m, 2 H), 7.59 (s, 2 H), 8.26 (s, 1 H). HPLC purity:
94.79%; HRMS [M + H] for C30H34N7O2, calcd, 524.2768; found,
524.2767.
(R,E)-3-(4-Amino-1-(1-(4-(piperazin-1-yl)but-2-enoyl)piperidin-3-
yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylphenyl)-
benzamide (60). To a solution of the Boc-protected piperazine
intermediate (prepared analogously to 59) (75 mg, 0.11, 1.0 equiv) in
dichloromethane (0.5 mL) was added 4 N HCl in dioxane (0.265 mL,
1.06 mmol, 10.0 equiv), and the reaction was stirred overnight. LC-MS
indicated the reaction was completed. The reaction was concentrated
to a residue and diluted with DMSO and subjected to HPLC purifi-
(R,E)-3-(4-Amino-1-(1-but-2-enoylpiperidin-3-yl)-1H-pyrazolo-
[3,4-d]pyrimidin-3-yl)-N-(4-isopropylphenyl)benzamide (55). By
following a procedure similar to that for target 57, the free piperidine
prepared for the synthesis of 40 (75 mg, 0.16 mmol, 1.0 equiv) was
1
converted into 55 (7 mg, 8%). H NMR (400 MHz, DMSO-d6): δ =
1.20 (d, J = 6.59 Hz, 6 H), 1.50−2.00 (m, 5 H), 2.13 (d, J = 8.79 Hz,
1 H), 2.22−2.36 (m, 1 H), 2.87 (dt, J = 13.73, 6.97 Hz, 1 H), 3.13−
3.25 (m, 1 H), 4.01−4.41 (m, 2 H), 4.46−4.85 (m, 2 H), 6.25−6.83
(m, 2 H), 7.23 (d, J = 8.79 Hz, 2 H), 7.61−7.76 (m, 3 H), 7.86 (d, J =
7.47 Hz, 1 H), 8.05 (d, J = 7.91 Hz, 1 H), 8.23 (br. s., 1 H), 8.29
(br. s., 1 H), 10.27 (s, 1 H). LC-MS: 100%, m/z = 524.3 [M + H];
HRMS [M + H] for C30H34N7O2, calcd, 524.2768; found, 524.2769.
(R)-3-(4-Amino-1-(1-methacryloylpiperidin-3-yl)-1H-pyrazolo-
[3,4-d]pyrimidin-3-yl)-N-(4-isopropylphenyl)benzamide (56). By
following a procedure similar to that for target 57, the free piperidine
prepared for the synthesis of 40 (75 mg, 0.16 mmol, 1.0 equiv) was
1
cation to provide the desired material as a solid (50 mg, 78%). H
NMR (600 MHz, DMSO-d6): δ = 1.12−1.31 (m, 6 H) 1.59 (br. s., 2
H) 1.95 (br. s., 2 H) 2.09−2.35 (m, 6 H) 2.59−2.76 (m, 4 H) 2.83−
3.17 (m, 4 H) 4.00−4.17 (m, 1 H) 4.50−4.83 (m, 1 H) 6.41−6.68 (m,
1 H) 7.23 (d, J = 8.33 Hz, 2 H) 7.58−7.76 (m, 3 H) 7.86 (d, J =
7.02 Hz, 1 H) 8.05 (d, J = 7.89 Hz, 1 H) 8.20−8.31 (m, 2 H) 10.23
(s, 1 H). LC-MS: 91%, m/z = 609.3 [M + H]; HRMS [M + Na] for
C34H41N9NaO2, calcd, 630.3275; found, 630.3275.
(R,E)-3-(4-Amino-1-(1-(4-(4-(2-aminoethyl)piperazin-1-yl)but-2-
enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-
isopropylphenyl)benzamide (61). Step 1: To a cooled solution of the
free piperidine prepared for the synthesis of 40 (500 mg, 1.10 mmol,
1.0 equiv) in dichloromethane (10 mL) at 0 °C was added tri-
ethylamine (0.305 mL, 2.20 mmol, 2.0 equiv). The reaction was stirred
for 5 min resulting in a homogeneous solution. To the reaction was
added (E)-4-bromobut-2-enoyl chloride (201 mg, 1.1 mmol, 1.0
equiv). The reaction was stirred for 1 h. The reaction was judged
complete by LC-MS and quenched with water and extracted with ethyl
acetate. The organic layer was then concentrated and purified via silica
gel chromatography (50% ethyl acetate in heptane to 100%) to pro-
vide the desired product as a solid (325 mg, 49%). LC-MS: 96%, m/z =
602.2/604.2 [M + H].
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converted into 56 (11 mg, 13%). H NMR (600 MHz, DMSO-d6):
δ = 1.20 (d, J = 7.02 Hz, 6 H), 1.58−1.65 (m, 2 H), 1.85 (br. s., 3 H),
1.94 (d, J = 13.59 Hz, 2 H), 2.12−2.20 (m, 2 H), 2.31 (d, J = 8.77 Hz,
1 H), 2.87 (dt, J = 13.70, 6.96 Hz, 1 H), 4.73−4.82 (m, 1 H), 4.96−
5.20 (m, 2 H), 7.22 (d, J = 8.33 Hz, 1 H), 7.65−7.72 (m, 3 H), 7.86
(d, J = 7.45 Hz, 1 H), 8.05 (d, J = 7.89 Hz, 1 H), 8.23 (s, 1 H), 8.28
(s, 1 H), 10.22 (s, 3 H). LC-MS: 100%, m/z = 524.3 [M + H]; HRMS
[M + H] for C30H34N7O2, calcd, 524.2768; found, 524.2770.
(R)-3-(4-Amino-1-(1-(3-methylbut-2-enoyl)piperidin-3-yl)-1H-
pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylphenyl)benzamide
(57). To a cooled solution of the free piperidine prepared for the
synthesis of 40 (75 mg, 0.16 mmol, 1.0 equiv) in dichloromethane
(4 mL) at 0 °C was added triethylamine (0.115 mL, 0.825 mmol,
5.0 equiv). The reaction was stirred for 5 min resulting in a homo-
geneous solution. To the reaction was added a solution of 3-methylcrotonoyl
chloride (0.018 mL, 0.165 mmol, 1.0 equiv) in dichloromethane (1 mL).
Step 2: To a solution of the bromocrotonamide from the previous
reaction (300 mg, 0.498 mmol, 1.0 equiv) in THF (10 mL) was added
triethylamine (0.208 mL, 1.49 mmol, 3.0 equiv) followed by tert-butyl
2-(piperazin-1-yl)ethylcarbamate (114 mg, 0.498 mmol, 1.0 equiv).
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dx.doi.org/10.1021/jm301190s | J. Med. Chem. 2012, 55, 10047−10063