Table S1. Potency of BMS-690514 in enzymatic kinase assays Kinase IC50, nmol/L EGFR 5 HER2 19 HER4 60 VEGFR1 50 VEGFR2 50 VEGFR3 25 IGF-1R >50,000 FLT3 110 Lck 220 FES 900 EMT >5000 SYK 4300 JAK3 5000 Cdk2/cyclin E 10000 PKA 280 PKCα 460 CAMKII 170 Akt1 >5000 Mek1 >10000 GSK3β >5000 IKKβ >10000 Table S2. KinomeScan Profile of BMS-690514 Target % control at 1 µM* AAK1 59 ABL1 4.8 ACK 14 ACTR2 100 ACTR2B 18 ADCK3 64 ADCK4 78 AKT1 68 AKT2 100 AKT3 78 ALK 100 ALK1 100 ALK2 86 ALK4 37 AMPKA1 87 AMPKA2 100 ANKRD3 56 ARG 1.4 AURA 100 AURB 100 AURC 100 AXL 100 BIKE 89 BLK 1.4 BMPR1A 78 BMPR1B 11 BMPR2 90 BMX 35 BRAF 100 BRK 20 BRSK1 100 BRSK2 57 BTK 69 CAMK1A 71 CAMK1D 56 CAMK1G 81 CAMK2A 53 CAMK2B 67 CAMK2D 46 CAMK2G 87 CAMK4 100 CAMKK1 100 CAMKK2 100 CAMLCK 100 CDC2L1_ISO4 100 CDC2L2_ISO1 100 CDK11 100 CDK2 100 CDK3 100 CDK5 100 CDK7 35 CDK8 100 CDK9 98 CDKL2 100 CHK1 70 CHK2 78 CK1A2 53 CK1D 52 CK1E 54 CK1G1 75 CK1G2 100 CK1G3 97 CK2A1 100 CK2A2 100 CLIK1 100 CLK1 72 CLK2 59 CLK3 67 CLK4 84 CRIK 34 CSK 48 DAPK1 79 DAPK2 89 DAPK3 68 DCAMKL1 100 DCAMKL2 100 DCAMKL3 100 DDR1 18 DDR2 39 DLK 91 DMPK1 52 DMPK2 100 DRAK1 100 DRAK2 100 DYRK1B 72 EGFR 0 EPHA1 8.1 EPHA2 44 EPHA3 31 EPHA4 48 EPHA5 93 EPHA6 73 EPHA7 100 EPHA8 24 EPHB1 53 EPHB2 76 EPHB3 27 EPHB4 50 ERK1 100 ERK2 86 ERK3 100 ERK4 100 ERK5 100 ERK7 93 FAK 100 FER 79 FES 29 FGFR1 72 FGFR2 100 FGFR3 100 FGFR4 100 FGR 1.3 FLT1 100 FLT3 37 FLT4 97 FMS 21 FRK 6.2 FUSED 80 FYN 4.5 GAK 1.9 GCK 94 GSK3A 36 GSK3B 87 HCK 3 HH498 7.8 HIPK1 100 HPK1 77 IGF1R 100 IKKA 70 IKKB 83 IKKE 90 INSR 100 IRAK3 57 IRR 100 ITK 100 JAK1 26 JAK1_PSEUDO 100 JAK2 17 JAK3_D2 41 JNK1 86 JNK2 80 JNK3 92 KHS1 54 KHS2 73 KIT 21 LATS1 100 LATS2 66 LCK 0.5 LIMK1 100 LIMK2 100 LKB1 70 LOK 9.1 LTK 56 LYN 5.6 MAP2K1 71 MAP2K2 57 MAP2K3 66 MAP2K4 72 MAP2K6 78 MAP3K3 0.1 MAP3K4 100 MAP3K5 100 MAPKAPK2 100 MAPKAPK5 97 MARK1 72 MARK2 91 MARK3 74 MARK4 76 MELK 100 MER 90 MET 100 MLK1 100 MLK2 91 MLK3 82 MNK1 100 MNK2 71 MRCKA 84 MRCKB 100 MSK1_C 72 MSK1_N 100 MSK2_C 36 MSK2_N 100 MSSK1 100 MST1 97 MST2 69 MST3 84 MST4 55 MUSK 90 MYO3A 77 MYO3B 77 MYT1 84 NDR2 70 NEK1 72 NEK2 69 NEK5 100 NEK6 100 NEK7 100 NEK9 100 NLK 100 NUAK1 44 NUAK2 32 P38A 83 P38B 100 P38D 100 P38G 80 PAK1 40 PAK2 69 PAK3 100 PAK4 100 PAK5 82 PAK6 75 PCTAIRE1 100 PCTAIRE2 100 PCTAIRE3 100 PDGFRA 81 PDGFRB 10 PDK1 82 PFTAIRE1 100 PFTAIRE2 68 PHKG1 61 PHKG2 77 PI3KC2B 68 PI3KCA 67 PI3KCB 90 PI3KCD 87 PI3KCG 85 PIM1 71 PIM2 49 PIM3 57 PIP5K1A 66 PIP5K2B 86 PKACA 6.9 PKACB 17 PKCD 16 PKCE 20 PKCH 36 PKCT 60 PKD1 2.8 PKD2 4.4 PKD3 74 PKG1 69 PKG2 55 PKN1 26 PKN2 20 PKR 84 PLK1 90 PLK3 100 PLK4 87 PRKX 5.2 PYK2 100 QIK 15 RAF1 80 RET 1.7 RIOK1 88 RIOK2 84 RIOK3 97 RIPK1 100 RIPK2 76 ROCK2 19 RON 91 ROS 46 RSK1_C 81 RSK1_N 93 RSK2_N 100 RSK3_C 77 RSK3_N 100 RSK4_C 80 RSK4_N 89 SGK085 100 SGK110 83 SGK288 82 SIK 16 SKMLCK 93 SLK 6.8 SMMLCK 95 SRC 0.2 SRM 82 SRPK1 98 SRPK2 100 STK16 97 SYK 62 TAK1 100 TAO1 59 TAO3 62 TEC 100 TESK1 90 TGFBR1 43 TGFBR2 18 TIE1 100 TIE2 100 TLK1 89 TLK2 73 TNK1 48 TRKA 100 TRKB 87 TRKC 90 TSSK1 100 TTK 85 TXK 2.4 TYK2 98 TYK2_PSEUDO 100 TYRO3 88 ULK1 67 ULK2 70 ULK3 84 WEE1 100 WEE1B 100 YANK2 88 YANK3 100 YES 9.2 YSK1 86 ZAK 43 ZAP70 99 ZC1/HGK 50 ZC2/TNIK 72 ZC3/MINK 5.3 * Data shown are percent binding of each kinase to its respective ligand in the presence of 1 µM BMS-690514, relative to binding with no competitor added. Table S3. Potency of BMS-690514 and Lapatinib in inhibiting HER2 and HER4 Phosphorylation IC50 in receptor phosphorylation Lapatinib BMS-690514 HER2 50 nM 60 nM HER4 wild type 730 nM 60 nM HER4 E317K 760 nM 70 nM HER4 R393W 860 nM 60 nM HER4 E452K 920 nM 100 nM Legends to Supplemental Figures. Fig. S1. Chemical structure of BMS-690514. Fig. S2. Fig. S2. BMS-690514 stabilizes the formation of inactive EGFR/HER2 heterodimers in the breast tumor cell line HCC1187. HCC1187 cells were treated with BMS-690514 for 1 h before stimulation with EGF for 5 min. Cell lysates were immunoprecipitated with antibodies specific for EGFR or HER2 and the immunoprecipitates (IP) were subjected to western blot analyses with antibodies to phosphotyrosine (pTyr), EGFR, or HER2. Fig. S3. Efficacy of BMS-690514 in EGFR- and HER2-dependent xenograft models. (A) Athymic mice implanted with PC9 non-small-cell lung tumors, which express an EGFR with an exon 19 deletion, were treated with vehicle only (), erlotinib at 100 mg/kg (), BMS-690514 at 30 mg/kg () or 60 mg/kg (). All treatments were given by oral gavage, once daily for 14 days. (B) Mice were implanted with N87 gastric tumors, which have HER2 gene amplification, and were treated for 21 days with vehicle (■), BMS-690514 at 90 mg/kg (◊), 60 mg/kg (), 30 mg/kg () or 15 mg/kg (Δ). The data represent the means ± standard errors of tumor sizes from 8 animals in each group. Fig. S4. Weight change in tumor-bearing mice treated with BMS-690514 (30 mg/kg, open circles; 60 mg/kg, filled squares) or erlotinib (100 mg/kg, open triangles) Fig. S5. Poly(ADP Ribose) Polymerase (PARP) cleavage in lung tumor cells treate with BMS-690514 for 24h. Whole-cell lysates were prepared and subjected to immunoblot analyses with antibody to PARP. The location of the cleavage product is indicated by an asterisk. Fig. S6. Plasma concentrations of BMS-6990514 in nude mice after a single dose at 3.75 mg/kg (open squares) or 7.5 mg/kg (filled squares). .