Reaction Steps
A solution of {1-[(5-methylisoxazol-3-yl)carbonyl]piperidin-4-yl}acetic acid (4.8 mg, 0.019 mmol) (prepared in Example C49, step I), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (19 mg, 0.050 mmol) in N,N-dimethylformamide (0.2 mL) and N,N-diisopropylethylamine (7.6 uL) was stirred for 15 minutes. A solution of 6-methyl-2,4,8,18,22-pentaazatetracyclo[14.3.1.1(3,7).1(9,13)]docosa-1(20),3(22),4,6,9(21),10,12,16,18-nonaen-12-amine tris(trifluoroacetate) (6.0 mg, 0.019 mmol) (prepared in Example C29, step C) in N,N-dimethylformamide (0.1 mL) and was added to the previous solution and was stirred overnight. The reaction mixture was purified by preparative HPLC (pH 2) to give the desired product (0.5 mg, 19%). LCMS for C30H33N8O3 (M+H)+: m/z=553.2. 1H NMR (300 MHz, DMSO-d6): δ 9.83 (s, 1H), 9.38 (s, 1H), 9.34 (s, 1H), 8.52 (s, 1H), 8.19 (s, 1H), 8.12 (s, 1H), 7.90 (s, 1H), 7.62 (d, J=2.4 Hz, 1H), 7.28 (d, J=8.7 Hz, 1H), 7.00 (dd, J=8.3, 2.2 Hz, 1H), 6.41 (dd, J=8.1, 0.8 Hz, 1H), 4.42 (m, 4H), 3.86 (m, 4H), 2.90-3.10 (m, 6H), 2.10-2.30 (m, 4H), 2.66 (s, 3H).
