Reaction SMILES: [CH2:1]([N:3]1[C:7]2=[N:8][C:9]([CH2:30][CH3:31])=[C:10]([CH2:19][NH:20][C:21]([CH:23]([CH2:27][CH2:28][CH3:29])[C:24](O)=[O:25])=[O:22])[C:11]([NH:12][CH:13]3[CH2:18][CH2:17][O:16][CH2:15][CH2:14]3)=[C:6]2[CH:5]=[N:4]1)[CH3:2].[NH2:32][CH2:33][C:34]1[CH:35]=[CH:36][C:37]([F:61])=[C:38]([C:40]2[CH:45]=[CH:44][CH:43]=[C:42]([CH2:46][N:47]3[CH2:52][CH2:51][N:50]([C:53]([O:55][C:56]([CH3:59])([CH3:58])[CH3:57])=[O:54])[C@@H:49]([CH3:60])[CH2:48]3)[CH:41]=2)[CH:39]=1.CN(C(ON1N=NC2C=CC=CC1=2)=[N+](C)C)C.F[P-](F)(F)(F)(F)F.CCN(CC)CC>C(Cl)Cl>[CH2:1]([N:3]1[C:7]2=[N:8][C:9]([CH2:30][CH3:31])=[C:10]([CH2:19][NH:20][C:21]([CH:23]([CH2:27][CH2:28][CH3:29])[C:24]([NH:32][CH2:33][C:34]3[CH:35]=[CH:36][C:37]([F:61])=[C:38]([C:40]4[CH:45]=[CH:44][CH:43]=[C:42]([CH2:46][N:47]5[CH2:52][CH2:51][N:50]([C:53]([O:55][C:56]([CH3:58])([CH3:57])[CH3:59])=[O:54])[C@@H:49]([CH3:60])[CH2:48]5)[CH:41]=4)[CH:39]=3)=[O:25])=[O:22])[C:11]([NH:12][CH:13]3[CH2:14][CH2:15][O:16][CH2:17][CH2:18]3)=[C:6]2[CH:5]=[N:4]1)[CH3:2]
Reactant: CCCC(C(=O)O)C(=O)NCc1c(CC)nc2c(cnn2CC)c1NC1CCOCC1
Reactant: C[C@H]1CN(Cc2cccc(-c3cc(CN)ccc3F)c2)CCN1C(=O)OC(C)(C)C
Agent: CN(C)C(On1nnc2ccccc21)=[N+](C)C
Agent: F[P-](F)(F)(F)(F)F
Solvent: ClCCl
Solvent: CCN(CC)CC
Product: CCCC(C(=O)NCc1ccc(F)c(-c2cccc(CN3CCN(C(=O)OC(C)(C)C)[C@@H](C)C3)c2)c1)C(=O)NCc1c(CC)nc2c(cnn2CC)c1NC1CCOCC1
Temperature: 25 °C
Reaction time: 8 h

Procedure:
A mixture of 2-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]pentanoic acid (120 mg, 0.278 mmol), 1,1-dimethylethyl (2S)-4-{[5′-(aminomethyl)-2′-fluoro-3-biphenylyl]methyl}-2-methyl-1-piperazinecarboxylate (115 mg, 0.278 mmol), HBTU (105 mg, 0.278 mmol) and Et3N (0.039 mL, 0.278 mmol) in DCM (3 mL) was stirred at room temperature overnight. The reaction was quenched with saturated NaHCO3 and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give crude residue. The crude product was purified with a Gilson HPLC (with 1% TFA in the solvent) eluting with 10 to 70% CH3CN in water at a flow rate of 20 mL/min. The product fractions were dried using a EZ2 GeneVac evaporator, combined and concentrated to give the intermediate 1,1-dimethylethyl (2S)-4-({5′-[({2-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]pentanoyl}amino)methyl]-2′-fluoro-3-biphenylyl}methyl)-2-methyl-1-piperazinecarboxylate. It was re-dissolved in 25% TFA in DCM (2 ML) and stirred for 2 h. Solvent was evaporated under a stream of nitrogen and then purified with a Gilson HPLC (with 0.1% TFA in the solvents), eluting with 10 to 70% CH3CN in water at a flow rate of 20 mL/min. The product fractions were combined, washed with saturated NaHCO3 and extracted with ethyl acetate twice. The combined organic layers were washed with brine, dried over sodium sulfate, filtered and then concentrated under vacuum to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-2-propylpropanediamide as a solid (53 mg, 26.2%). LC-MS m/z 727 (M+H)+, 0.72 min (ret time).