Supporting Information The design of potent and selective inhibitors to overcome clinical ALK mutations resistant to crizotinib Qinhua Huang,* Ted W. Johnson,* Simon Bailey, Alexei Brooun, Kevin D. Bunker, Benjamin J. Burke, Michael R. Collins, Andrew S. Cook, J. Jean Cui, Kevin N. Dack, Judith G. Deal, Ya-Li Deng, Dac Dinh, Lars D. Engstrom, Mingying He, Jacqui Hoffman, Robert L. Hoffman, Patrick S. Johnson, Robert S. Kania, Hieu Lam, Justine L. Lam, Phuong T. Le, Qiuhua Li, Laura Lingardo, Wei Liu, Melissa West Lu, Michele McTigue, Cynthia L. Palmer, Paul F. Richardson, Neal W. Sach, Hong Shen, Tod Smeal, Graham L. Smith, Albert E. Stewart, Sergei Timofeevski, Konstantinos Tsaparikos, Hui Wang, Huichun Zhu, Jinjiang Zhu, Helen Y. Zou, Martin P. Edwards La Jolla Laboratories, Pfizer Worldwide Research and Development, 10770 Science Center Drive, San Diego, CA 92121, United States Contents Biochemical kinase selectivity for compound 8e S2-S8 S1
Biochemical kinase selectivity for compound 8e Kinase %Inhibition (Invitrogen) % Inhibition at 1µM % inhibition at 0.1µM IC 50 (nm) Selectivity ratio a ALK L1196M (Not in panel) (Not in panel) 0.2 (Ki b ) 1 FER 104 100 2 5 ROS1 103 99 0.02 (Ki b ) 0.1 PTK2 (FAK) 99 98 16 40 LTK (TYK1) 103 97 2 5 PTK2B (FAK2) 98 96 12 30 NTRK2 (TRKB) 106 95 4 10 NTRK3 (TRKC) 98 94 18 45 TNK2 (ACK) 100 94 15 38 JAK2 112 93 12 30 MUSK 100 91 31 78 JAK1 101 89 25 63 FES (FPS) 97 86 5 13 JAK2 JH1 JH2 V617F 92 85 32 80 EPHA1 98 85 28 70 JAK2 JH1 JH2 98 84 29 73 NTRK1 (TRKA) 89 83 17 42 EPHB1 95 81 37 92 LRRK2 83 81 57 142 EPHA7 100 80 46 114 IGF1R 95 79 46 115 EPHA2 93 67 53 132 TEK (Tie2) 95 67 104 260 INSR 95 65 65 163 MET (cmet) 94 63 45 112 EPHA4 93 58 EPHB4 91 58 ABL1 T315I 88 56 TYK2 90 55 EPHA5 87 53 TGFBR1 (ALK5) 90 52 MAP4K4 (ZC1) 94 33 289 723 S2
EPHB2 88 27 MERTK (cmer) 84 23 AURKA (AURA) 79 139 348 INSRR (IRR) 77 MET M1250T 76 CSF1R (FMS) 75 MAP3K2 (MEKK2) 74 TXK 73 ABL2 (Arg) 73 IRAK1 69 FRK (PTK5) 69 AXL 68 LCK 65 239 MAP3K3 (MEKK3) 64 JAK3 63 EGFR (ErbB1) T790M L858R 61 20 ABL1 Y253F 60 MST1R (RON) 60 MARK2 57 EPHA8 55 LYN A 55 FLT4 (VEGFR3) 55 LYN B 53 FGR 53 ABL1 G250E 53 EGFR (ErbB1) T790M 52 SRC N1 52 ABL1 51 BLK 49 DCAMKL2 (DCK2) 48 SRC 44 STK22B (TSSK2) 43 TAOK2 (TAO2) 42 MARK1 41 RIPK2 40 S3
PTK6 (Brk) 40 CDK1/cyclin B 39 KDR (VEGFR2) 38 BMX 37 MAP3K7/MAP3K7IP1 (TAK1-TAB1) 37 HCK 36 ABL1 E255K 35 YES1 33 CSK 31 MST4 31 EPHA3 30 EPHB3 30 FLT3 D835Y 28 CDK5/p35 27 BTK 26 ROCK1 (ROCKI) 25 FLT3 25 FRAP1 (mtor) 25 2370 EGFR (ErbB1) L861Q 24 SRMS (Srm) 23 TYRO3 (RSE) 23 EGFR (ErbB1) 22 CDK2 /CyclinA 22 CDK7/cyclin H/MNAT1 21 STK3 (MST2) 21 RAF1 (craf) Y340D Y341D 21 MAP2K6 (MKK6) 20 STK16 (PKL12) 20 ACVR1B (ALK4) 20 SYK 20 BRSK1 (SAD1) 19 PDGFRB (PDGFR beta) 18 DMPK 18 PDGFRA V561D 18 S4
PAK4 17 EGFR (ErbB1) L858R 17 PDGFRA T674I 17 LIMK1 16 MYLK2 (MLCK_sk) 16 FGFR1 14 HIPK4 14 BRAF V599E 13 DNA-PK 13 KIT T670I 13 PRKD1 (PKC mu) 12 ITK 12 FGFR3 12 CAMK2A (CaMKIIa) 12 CAMK2D (CaMKII delta) 12 MAP2K3 (MEK3) 12 PRKCE (PKC epsilon) 11 FYN 10 PDGFRA (PDGFR alpha) 10 PKN1 (PRK1) 10 PKN1 (PRK1) 10 GSK3A (GSK3 alpha) 9 TBK1 9 CHUK (IKK alpha) 9 MATK (HYL) 9 MAP3K8 (COT) 9 MAPK1 (ERK2) 9 CDK9/cyclin T1 8 RET 8 PAK6 8 CAMK1D (CaMKI delta) 7 PIM2 7 DAPK1 7 KIT V654A 7 TEC 7 S5
MAPK3 (ERK1) 7 PAK1 6 MAPK14 (p38) direct 6 PRKD2 (PKD2) 6 CSNK2A2 (CKIIa prime) 6 MAPKAPK2 (MK2) 6 BRAF 6 CLK1 6 PRKG1 5 CAMK4 (CaMKIV) 5 MAP2K2 (MEK2) 5 CAMKK1 (CAMKKA) 5 RET V804L 5 PLK2 5 STK33 5 PASK 4 PRKCZ (PKC zeta) 4 RPS6KA3 (RSK2) 4 MAPK10 (JNK3) 4 CHEK2 (CHK2) 4 NEK7 4 CAMK2B (CaMKII beta) 4 WEE1 3 DDR1 3 MAP3K10 (MLK2) 3 FGFR3 K650E 3 ZAP70 3 NEK2 3 AKT3 (PKB gamma) 3 AKT1 (AKT) 3 PRKCB1 (PKC beta I) 2 PAK2 (PAK65) 2 RPS6KA1 (RSK1) 2 DDR2 2 PLK3 2 CHEK1 (CHK1) 2 S6
ERBB4 (HER4) 2 PRKCG (PKC gamma) 2 CDC42 BPB (MRCKB) 1 RET Y791F 1 CDK8/cyclin C 1 GSG2 (Haspin) 1 IKBKB (IKKb) 1 STK17A (DRAK1) 1 AMPK A2/B1/G1 1 DYRK3 1 CSNK1A1 (CKIa) 1 MAP3K14 (NIK) 1 PRKACA (PKACa) 0 PDPK1 (PDK1) (direct) 0 FGFR4 0 SGK1 (SGK) 0 EEF2K 0 DAPK3 (ZIPK) 0 SGKL (SGK3) -1 ERBB2 (HER2) -1 ZAK -1 KIT -1 FLT1 (VEGFR1) -1 MAPKAPK3-1 PDGFRA D842V -2 PRKCD (PKC delta) -2 RPS6KA4 (MSK2) -3 RPS6KA4 (MSK2) -3 FGFR2-3 WNK2-3 PRKCQ (PKC theta) -3 CSNK1E (CK1 epsilon) -4 GRK4-4 DYRK1B -5 MAP3K5 (ASK1) -19 S7
a Invitrogen Z -LYTEenzyme IC 50 using K m -levels of ATP except where noted differently. MAP3K8 (COT), MAPK10 (JNK3) and RAF1 (craf) Y340D Y341D were tested by Z -LYTEcascade assays with 100uM ATP. CDK7/cyclin H/MNAT1, CDK9/cyclin T1, CHUK (IKK alpha), DAPK1, GSG2 (Haspin), IRAK1, LRRK2 were tested by Adapta assays using Km-level of ATP. BRAF, BRAF V599E, CAMKK1 (CAMKKA), CDK8/cyclin C, DDR1, DDR2, DMPK, LIMK1, MAP2K2 (MEK2), MAP2K3 (MEK3), MAP2K6 (MKK6), MAP3K10 (MLK2), MAP3K14 (NIK), MAP3K2 (MEKK2), MAP3K3 (MEKK3), MAP3K5 (ASK1), MAP3K7/MAP3K7IP1 (TAK1- TAB1), RIPK2, STK16 (PKL12), STK17A (DRAK1), STK33, TEC, TGFBR1 (ALK5), TNK2 (ACK), WEE1, WNK2, ZAK were tested by LanthaScreen binding assays. K i = ~ IC 50 /2 from the Cheng-Prusoff equation, K i = IC 50 /(1 + [ATP]/K m ), for competitive inhibition. b K i values calculated using tight-binding (Morrison) equation for competitive inhibitors tested in a Pfizer mobility shift assay. S8