Supplemental Data Table 1. R406 Solubility in a Range of Buffers

Supplemental data table 1. R406 solubility in a range of buffers.

8 representative buffers were selected from the >200 individual buffers used in the pharmacological profiling experiments. Solubility was determined as described in the Methods section, at 3 hour and 24 hour timepoints with a range of DMSO concentrations.

DTT – dithiothreitol. BSA – bovine serum albumin. PBS – phosphate-buffered saline.

R406 solubility (µM) 3 hour timepoint 24 hour timepoint Buffer name Buffer composition 0.1% 1% DMSO DMSO 0.1% DMSO 0.3% DMSO 1% DMSO Non-kinase buffer 1 20 mM HEPES, 100 mM NaCl, 10 mM 0.8 1.4 0.4 2.5 MgCl2, 1 mM DTT, 1 mM EDTA, pH 7.4 Non-kinase buffer 2 25 mM HEPES, pH 7.4, 5 mM MgCl2, 1 12.1 5.6 3.5 2.4 mM CaCl2, 0.1% BSA Non-kinase buffer 3 50 mM Tris-HCl, pH 7.5, 8 mM MgCl2, 25 µg mL-1 alamethicin, 2 mM UDPGA, 8.8 1.8 2.6 1.4 0.25% methanol Kinase binding buffer 20% SeaBlock, 0.17x PBS, 0.05% Tween 26.5 219.8 29.1 92.3 192.4 20, 6 mM DTT Isolated kinase buffer 1 20 mM MOPS, 1 mM EDTA, 0.01% Brij- 35, 5% Glycerol, 0.1% - 9.8 9.5 11.2 10.5 mercaptoethanol, 1 mg mL-1 BSA Isolated kinase buffer 2 50 mM TRIS, 0.1mM EGTA, 0.1% - 14.3 3 1.7 1.9 mercaptoethanol, 1 mg mL-1 BSA Isolated kinase buffer 3 20 mM HEPES, 0.03% Triton X-100 41 169.8 38.6 57.1 121.2 Phosphate-buffered PBS, pH 7.4, no BSA 0.8 1.1 2.9 1.3 saline Supplemental data table 2. Full in vitro pharmacological profile of R406

In vitro pharmacological profiling results for R406 in radioligand binding and isolated enzyme activity assays, at all target classes tested other than kinases.

Target class and Target subclass are provided to indicate the breadth of coverage. pKi values are presented for radioligand binding assays only. A pIC50 value of 3.5 indicates inactivity in the assay (defined as <50% inhibition in the initial 10 µM single concentration test, or activity in this test not replicated in the concentration-response curve). In a small number of cases, multiple assays have been used for the same target (e.g. 3 different binding sites on the L-type calcium channel); these assays have a common target but different assay name. Assay name and Target do not necessarily correspond to individual gene or protein identifiers since many targets are composed of multiple protein subunits and as such cannot be identified by a single gene or protein identifier.

Target Assay name Target class pIC50 pKi Acetylcholinesterase Acetylcholinesterase Enzyme 3.5 Acyl CoA-cholesterol acyltransferase, Acyl CoA-Cholesterol Acyltransferase, intestine Intestine Enzyme 3.5 Acyl CoA-cholesterol acyltransferase, Acyl CoA-Cholesterol Acyltransferase, hepatic Hepatic Enzyme 3.5

Adenosine A1 receptor Adenosine A1 receptor Receptor 3.5

Adenosine A2A receptor Adenosine A2A receptor Receptor 3.5

Adenosine A2B receptor Adenosine A2B receptor Receptor 3.5

Adenosine A3 receptor Adenosine A3 receptor Receptor 7.7 7.8

1A adrenoceptor 1A adrenoceptor Receptor 3.5

1B adrenoceptor 1B adrenoceptor Receptor 3.5

1D adrenoceptor 1D adrenoceptor Receptor 3.5

2A adrenoceptor 2A adrenoceptor Receptor 3.5

2B adrenoceptor 2B adrenoceptor Receptor 3.5

2C adrenoceptor 2C adrenoceptor Receptor 3.5

1 adrenoceptor 1 adrenoceptor Receptor 3.5

2 adrenoceptor 2 adrenoceptor Receptor 3.5 3 adrenoceptor 3 adrenoceptor Receptor 3.5

Adrenomedullin AM1 receptor Adrenomedullin AM1 receptor Receptor 3.5

Adrenomedullin AM2 receptor Adrenomedullin AM2 receptor Receptor 3.5 Aldehyde dehydrogenase, mitochondrial Aldehyde dehydrogenase Enzyme 3.5 Aldose reductase Aldose reductase Enzyme 3.5 Aldosterone receptor Aldosterone receptor Receptor 3.5 -D-Glucosidase -D-Glucosidase Enzyme 3.5 Anaphylatoxin C5a receptor Anaphylatoxin C5a receptor Receptor 3.5

Angiotensin AT1 receptor Angiotensin AT1 receptor Receptor 3.5

Angiotensin AT2 receptor Angiotensin AT2 receptor Receptor 3.5 Apelin APJ receptor Apelin APJ receptor Receptor 3.5 ATPase, Ca2+, skeletal muscle, pig ATPase, Ca2+ Enzyme 3.5 ATPase, H+/K+, stomach, pig ATPase, H+/K+ Enzyme 3.5 ATPase, Na+/K+, pig heart ATPase, Na+/K+ Enzyme 3.5 Atrial natriuretic factor receptor (ANF) Atrial natriuretic factor receptor (ANF) Receptor 3.5 Benzodiazepine receptor, peripheral Benzodiazepine receptor, peripheral Transporter 3.5

Bombesin BB1 receptor Bombesin BB1 receptor Receptor 3.5

Bradykinin B1 receptor Bradykinin B1 receptor Receptor 3.5

Bradykinin B2 receptor Bradykinin B2 receptor Receptor 3.5 Calcitonin receptor Calcitonin receptor Receptor 3.5 Calcitonin gene-related peptide receptor Calcitonin gene-related peptide receptor CGRP1 CGRP1 Receptor 3.5 L-type calcium channel, benzothiazepine site L-type calcium channel Ion channel 3.5 L-type calcium channel, dihydropyridine site L-type calcium channel Ion channel 3.5 L-type calcium channel, phenylalkylamine site L-type calcium channel Ion channel 3.5 N-type calcium channel N-type calcium channel Ion channel 3.5

Cannabinoid CB1 receptor Cannabinoid CB1 receptor Receptor 3.5

Cannabinoid CB2 receptor Cannabinoid CB2 receptor Receptor 3.5 Carbonic Anhydrase II Carbonic Anhydrase II Enzyme 3.5 Carnitine Palmitoyltransferase-1 (CPT-1) Carnitine Palmitoyltransferase-1 (CPT-1) Enzyme 3.5 Catechol-O-Methyltransferase (COMT) Catechol-O-Methyltransferase (COMT) Enzyme 3.5 Chemokine CCR1 receptor Chemokine CCR1 receptor Receptor 3.5 Chemokine CCR2B receptor Chemokine CCR2B receptor Receptor 3.5 Chemokine CCR4 receptor Chemokine CCR4 receptor Receptor 3.5 Chemokine CCR5 receptor Chemokine CCR5 receptor Receptor 3.5

Chemokine CX3CR1 receptor Chemokine CX3CR1 receptor Receptor 3.5 Chemokine CXCR1/2 receptor (IL-8, Non- Chemokine CXCR1/2 receptor (IL-8, Non- Selective) Selective) Receptor 3.5 Chemokine CXCR2 receptor (IL-8B) Chemokine CXCR2 receptor (IL-8B) Receptor 3.5 Chemokine CXCR4 receptor Chemokine CXCR4 receptor Receptor 3.5

Cholecystokinin CCK1 receptor Cholecystokinin CCK1 receptor Receptor 3.5

Cholecystokinin CCK2 receptor Cholecystokinin CCK2 receptor Receptor 3.5 Cholinesterase, butyryl Cholinesterase, butyryl Enzyme 3.5 Corticotropin releasing factor receptor Corticotropin releasing factor receptor Receptor 3.5 Cyclooxygenase COX-1 Cyclooxygenase COX-1 Enzyme 3.5 Cyclooxygenase COX-2 Cyclooxygenase COX-2 Enzyme 3.5 Deacetylase, histone Deacetylase, histone Enzyme 3.5 Deacetylase, sirtuin SIRT1 Deacetylase, sirtuin SIRT1 Enzyme 3.5 Deacetylase, sirtuin SIRT2 Deacetylase, sirtuin SIRT2 Enzyme 3.5 Deacetylase, sirtuin SIRT3 Deacetylase, sirtuin SIRT3 Enzyme 3.5

Dopamine D1 receptor Dopamine D1 receptor Receptor 3.5

Dopamine D2L receptor Dopamine D2L receptor Receptor 3.5

Dopamine D2S receptor Dopamine D2S receptor Receptor 3.5

Dopamine D4.2 receptor Dopamine D4.2 receptor Receptor 3.5

Dopamine D4.4 receptor Dopamine D4.4 receptor Receptor 3.5

Endothelin ETA receptor Endothelin ETA receptor Receptor 3.5

Endothelin ETB receptor Endothelin ETB receptor Receptor 3.5 Epidermal Growth Factor Receptor receptor Epidermal Growth Factor Receptor receptor (EGF) (EGF) Enzyme 3.5 Estrogen ER receptor Estrogen ER receptor Receptor 3.5 Estrogen ER receptor Estrogen ER receptor Receptor 3.5 Fatty acid amide hydrolase (FAAH) Fatty acid amide hydrolase (FAAH) Enzyme 5.8 G protein-coupled receptor GPR103 G protein-coupled receptor GPR103 Receptor 3.5 G protein-coupled eeceptor GPR54 (KiSS1) G protein-coupled eeceptor GPR54 (KiSS1) Receptor 3.5 G protein-coupled receptor GPR8 G protein-coupled receptor GPR8 Receptor 3.5

GABAA, agonist site GABAA, agonist site Ion channel 3.5

GABAA, benzodiazepine, central GABAA, benzodiazepine, central Ion channel 3.5

GABAA, chloride channel, TBOB GABAA, chloride channel, TBOB Ion channel 3.5

GABAA, chloride channel, TBPS GABAA, chloride channel, TBPS Ion channel 3.5

GABAB1A receptor GABAB1A receptor Receptor 3.5 Gabapentin receptor Gabapentin receptor Receptor 3.5

Galanin GAL1 receptor Galanin GAL1 receptor Receptor 3.5

Galanin GAL2 receptor Galanin GAL2 receptor Receptor 3.5 γ-Hydroxybutyric acid (GHB) receptor gγ-Hydroxybutyric acid (GHB) receptor Receptor 3.5 Glucagon-Like Peptide-1 (GLP-1) receptor Glucagon-Like Peptide-1 (GLP-1) receptor Receptor 3.5 Glucocorticoid receptor Glucocorticoid receptor Receptor 3.5 Glutamate receptor, AMPA Glutamate receptor, AMPA Ion channel 3.5 Glutamate receptor, kainate Glutamate receptor, kainate Ion channel 3.5

Glutamate receptor, metabotropic, mGlu5 Glutamate receptor, metabotropic, mGlu5 Receptor 3.5 Glutamate receptor, NMDA, agonist site Glutamate receptor, NMDA, agonist site Ion channel 3.5 Glutamate receptor, NMDA, glycine site Glutamate receptor, NMDA, glycine site Ion channel 3.5 Glutamate receptor, NMDA, phencyclidine Glutamate receptor, NMDA, phencyclidine site site Ion channel 3.5 Glutamate receptor, NMDA, polyamine site Glutamate receptor, NMDA, polyamine site Ion channel 3.5 Glycine receptor, strychnine-sensitive Glycine receptor, strychnine-sensitive Ion channel 3.5 Glycogen phosphorylase Glycogen phosphorylase Enzyme 3.5 Growth hormone secretagogue (GHS, Growth hormone secretagogue (GHS, Ghrelin) receptor Ghrelin) receptor Receptor 3.5

Histamine H1 receptor Histamine H1 receptor Receptor 3.5

Histamine H4 receptor Histamine H4 receptor Receptor 3.5 Histamine N-methyltransferase (HNMT) Histamine N-methyltransferase (HNMT) Enzyme 3.5 HMG-CoA reductase HMG-CoA reductase Enzyme 3.5

Imidazoline I2 receptor, central Imidazoline I2 receptor, central Receptor 3.5

Imidazoline I2 receptor, peripheral Imidazoline I2 receptor, peripheral Receptor 3.5

Inositol trisphosphate IP3 receptor Inositol trisphosphate IP3 receptor Receptor 3.5 Insulin receptor Insulin receptor Enzyme 3.5 Interleukin IL-1 receptor Interleukin IL-1 receptor Receptor 3.5 Interleukin IL-2 receptor Interleukin IL-2 receptor Receptor 3.5 Interleukin IL-6 receptor Interleukin IL-6 receptor Receptor 3.5 Leptin receptor Leptin receptor Receptor 3.5

Leukotriene LTA4 hydrolase Leukotriene LTA4 hydrolase Enzyme 3.5

Leukotriene BLT1 receptor Leukotriene BLT1 receptor Receptor 3.5

Leukotriene LTC4 synthase Leukotriene LTC4 synthase Enzyme 3.5

Leukotriene, cysteinyl CysLT1 receptor Leukotriene, cysteinyl CysLT1 receptor Receptor 3.5 Lipase Lipase Enzyme 3.5 Lipid peroxidase Lipid peroxidase Enzyme 3.5 Lipoxygenase 12-LO Lipoxygenase 12-LO Enzyme 3.5 Lipoxygenase 15-LO Lipoxygenase 15-LO Enzyme 3.5 Lipoxygenase 5-LO Lipoxygenase 5-LO Enzyme 5.3

Melanocortin MC1 receptor Melanocortin MC1 receptor Receptor 3.5

Melanocortin MC5 receptor Melanocortin MC5 receptor Receptor 3.5 Melatonin ML (MT) receptor, non-selective Melatonin ML (MT) receptor, non-selective Receptor 3.5

Melatonin MT1 receptor Melatonin MT1 receptor Receptor 3.5 Monoamine oxidase MAO-A Monoamine oxidase MAO-A Enzyme 3.5 Monoamine oxidase MAO-B Monoamine oxidase MAO-B Enzyme 3.5 Motilin receptor Motilin receptor Receptor 3.5

Muscarinic M1 receptor Muscarinic M1 receptor Receptor 3.5

Muscarinic M5 receptor Muscarinic M5 receptor Receptor 3.5 Myeloperoxidase Myeloperoxidase Enzyme 3.5

Neuromedin U NMU1 receptor Neuromedin U NMU1 receptor Receptor 3.5

Neuromedin U NMU2 receptor Neuromedin U NMU2 receptor Receptor 3.5

Neuropeptide Y1 receptor Neuropeptide Y1 receptor Receptor 3.5

Neuropeptide Y2 receptor Neuropeptide Y2 receptor Receptor 3.5

Neurotensin NT1 receptor Neurotensin NT1 receptor Receptor 3.5 N-formyl peptide receptor FPR1 N-formyl peptide receptor FPR1 Receptor 3.5

N-formyl peptide receptor-like FPRL1 N-formyl peptide receptor-like FPRL1 Receptor 3.5 Nicotinic acetylcholine receptor Nicotinic acetylcholine receptor Ion channel 3.5

Nicotinic acetylcholine receptor 42, Nicotinic acetylcholine receptor 42, cytisine cytisine Ion channel 3.5

Nicotinic acetylcholine receptor 7, Nicotinic acetylcholine receptor 7, methyllycaconitine methyllycaconitine Ion channel 3.5 Nicotinic acetylcholine receptor, Nicotinic acetylcholine receptor, bungarotoxin-sensitive, central bungarotoxin-sensitive, central Ion channel 3.5 Nicotinic acetylcholine receptor, Nicotinic acetylcholine receptor, bungarotoxin-sensitive, neuromuscular bungarotoxin-sensitive, neuromuscular Ion channel 3.5 Nitric oxide synthase, endothelial (eNOS) Nitric oxide synthase, endothelial (eNOS) Enzyme 3.5 Nitric oxide synthase, inducible (iNOS) Nitric oxide synthase, inducible (iNOS) Enzyme 3.5 Nitric oxide synthase, neuronal (nNOS) Nitric oxide synthase, neuronal (nNOS) Enzyme 3.5 δ receptor (opioid) δ receptor (opioid) Receptor 3.5 κ receptor (opioid) κ receptor (opioid) Receptor 3.5 μ receptor (opioid) μ receptor (opioid) Receptor 3.5

Orexin OX1 receptor Orexin OX1 receptor Receptor 3.5

Orexin OX2 receptor Orexin OX2 receptor Receptor 3.5 NOP receptor (ORL1) NOP receptor (ORL1) Receptor 3.5 Matrix metalloproteinase-1 (MMP-1) Matrix metalloproteinase-1 (MMP-1) Enzyme 3.5 Matrix metalloproteinase-7 (MMP-7) Matrix metalloproteinase-7 (MMP-7) Enzyme 3.5 Angiotensin Converting Enzyme Angiotensin Converting Enzyme Enzyme 3.5 BACE1 (-Secretase) BACE1 (-Secretase) Enzyme 3.5 Calpain-1 Calpain-1 Enzyme 3.5 Caspase 1 Caspase 1 Enzyme 3.5 Caspase 10 Caspase 10 Enzyme 3.5 Caspase 2 Caspase 2 Enzyme 3.5 Caspase 3 Caspase 3 Enzyme 3.5 Caspase 4 Caspase 4 Enzyme 3.5 Caspase 5 Caspase 5 Enzyme 3.5 Caspase 6 Caspase 6 Enzyme 3.5 Caspase 7 Caspase 7 Enzyme 3.5 Caspase 8 Caspase 8 Enzyme 3.5 Caspase 9 Caspase 9 Enzyme 3.5 Cathepsin B Cathepsin B Enzyme 3.5 Cathepsin C Cathepsin C Enzyme 3.5 Cathepsin D Cathepsin D Enzyme 3.5 Cathepsin G Cathepsin G Enzyme 3.5 Cathepsin H Cathepsin H Enzyme 3.5 Cathepsin L Cathepsin L Enzyme 5.2 Chymase Chymase Enzyme 3.5 Chymotrypsin Chymotrypsin Enzyme 3.5 Cathepsin E Cathepsin E Enzyme 3.5 Cathepsin L2 Cathepsin L2 Enzyme 3.5 Cathepsin S Cathepsin S Enzyme 4.9 Cathepsin Z Cathepsin Z Enzyme 3.5 Dipeptidyl peptidase 4 (DPP4) Dipeptidyl peptidase 4 (DPP4) Enzyme 3.5 Pancreatic Elastase 1 (ELA1) Pancreatic Elastase 1 (ELA1) Enzyme 3.5 Neutrophil Elastase 2 (ELA2) Neutrophil Elastase 2 (ELA2) Enzyme 3.5 Endothelin converting enzyme-1 (ECE-1) Endothelin converting enzyme-1 (ECE-1) Enzyme 3.5 Factor VIIa Factor VIIa Enzyme 3.5 Factor Xa Factor Xa Enzyme 3.5 Kallikrein, plasma Kallikrein, plasma Enzyme 3.5 Matrix metalloproteinase-12 (MMP-12) Matrix metalloproteinase-12 (MMP-12) Enzyme 3.5 Matrix metalloproteinase-13 (MMP-13) Matrix metalloproteinase-13 (MMP-13) Enzyme 3.5 Matrix metalloproteinase-14 (MMP-14) Matrix metalloproteinase-14 (MMP-14) Enzyme 3.5 Matrix metalloproteinase-2 (MMP-2) Matrix metalloproteinase-2 (MMP-2) Enzyme 3.5 Matrix metalloproteinase-3 (MMP-3) Matrix metalloproteinase-3 (MMP-3) Enzyme 3.5 Matrix metalloproteinase-8 (MMP-8) Matrix metalloproteinase-8 (MMP-8) Enzyme 3.5 Matrix metalloproteinase-9 (MMP-9) Matrix metalloproteinase-9 (MMP-9) Enzyme 3.5 Metalloproteinase, neutral endopeptidase Metalloproteinase, neutral endopeptidase Enzyme 3.5 Pepsin Pepsin Enzyme 3.5 Plasmin Plasmin Enzyme 3.5 PLAU (Urokinase) PLAU (Urokinase) Enzyme 3.5 Prolyl oligopeptidase (POP) Prolyl oligopeptidase (POP) Enzyme 3.5 Renin Renin Enzyme 3.5 Thrombin Thrombin Enzyme 3.5 Tissue plasminogen activator (tPA) Tissue plasminogen activator (tPA) Enzyme 3.5 Trypsin Trypsin Enzyme 3.5 Tryptase Tryptase Enzyme 3.5 Tumor necrosis factor- converting enzyme Tumor necrosis factor- converting enzyme (TACE) (TACE) Enzyme 3.5 Phorbol ester receptor Phorbol ester receptor Receptor 3.5 Phosphodiesterase PDE1 Phosphodiesterase PDE1 Enzyme 3.5 Phosphodiesterase PDE10A1 Phosphodiesterase PDE10A1 Enzyme 3.5 Phosphodiesterase PDE2 Phosphodiesterase PDE2 Enzyme 3.5 Phosphodiesterase PDE3 Phosphodiesterase PDE3 Enzyme 3.5 Phosphodiesterase PDE4 Phosphodiesterase PDE4 Enzyme 3.5 Phosphodiesterase PDE5 Phosphodiesterase PDE5 Enzyme 6.5 Phosphodiesterase PDE6 Phosphodiesterase PDE6 Enzyme 3.5 Phospholipase PLA2-I Phospholipase PLA2-I Enzyme 3.5 Phospholipase PLA2-II Phospholipase PLA2-II Enzyme 3.5 Phospholipase PLC Phospholipase PLC Enzyme 3.5 Platelet activating factor (PAF) receptor Platelet activating factor (PAF) receptor Receptor 3.5 Platelet-derived growth factor receptor Platelet-derived growth factor receptor Enzyme 3.5 Potassium channel [KA] Potassium channel [KA] Ion channel 3.5 Potassium channel [KATP] Potassium channel [KATP] Ion channel 3.5 Potassium channel [SKCa] Potassium channel [SKCa] Ion channel 3.5 Progesterone receptor Progesterone receptor Receptor 3.5 Progesterone receptor PR-B Progesterone receptor PR-B Receptor 3.5

Prostanoid CRTH2 receptor Prostanoid CRTH2 receptor Receptor 3.5 Prostanoid DP receptor Prostanoid DP receptor Receptor 3.5

Prostanoid EP2 receptor Prostanoid EP2 receptor Receptor 3.5

Prostanoid EP4 receptor Prostanoid EP4 receptor Receptor 3.5 Prostanoid TP receptor Prostanoid TP receptor Receptor 3.5 Proteasome Proteasome Enzyme 3.5 Protein serine/threonine phosphatase, PP1 Protein serine/threonine phosphatase, PP1 Enzyme 3.5 Protein serine/threonine phosphatase, PP2A Protein serine/threonine phosphatase, PP2A Enzyme 3.5 Protein serine/threonine phosphatase, Protein serine/threonine phosphatase, PPM1A (PP2C-) PPM1A (PP2C-) Enzyme 3.5 Protein serine/threonine phosphatase, Protein serine/threonine phosphatase, Enzyme 3.5 PPP3CA (calcineurin, PP2B) PPP3CA (calcineurin, PP2B) Protein tyrosine phosphatase, ACP1 Protein tyrosine phosphatase, ACP1 (LMPTP-A) (LMPTP-A) Enzyme 3.5 Protein tyrosine phosphatase, ACP1 Protein tyrosine phosphatase, ACP1 (LMPTP-B) (LMPTP-B) Enzyme 3.5 Protein tyrosine phosphatase, CDC25A Protein tyrosine phosphatase, CDC25A Enzyme 3.5 Protein tyrosine phosphatase, CDC25B Protein tyrosine phosphatase, CDC25B Enzyme 3.5 Protein tyrosine phosphatase, CDC25C Protein tyrosine phosphatase, CDC25C Enzyme 3.5 Protein tyrosine phosphatase, DUSP1 (MKP- Protein tyrosine phosphatase, DUSP1 (MKP- 1, HVH1, CL100) 1, HVH1, CL100) Enzyme 3.5 Protein tyrosine phosphatase, DUSP10 Protein tyrosine phosphatase, DUSP10 (MKP-5) (MKP-5) Enzyme 3.5 Protein tyrosine phosphatase, DUSP14 Protein tyrosine phosphatase, DUSP14 (MKP-6) (MKP-6) Enzyme 3.5 Protein tyrosine phosphatase, DUSP3 (VHR) Protein tyrosine phosphatase, DUSP3 (VHR) Enzyme 3.5 Protein tyrosine phosphatase, DUSP4 (MKP- Protein tyrosine phosphatase, DUSP4 (MKP- 2, HVH2) 2, HVH2) Enzyme 3.5 Protein tyrosine phosphatase, DUSP6 (MKP- Protein tyrosine phosphatase, DUSP6 (MKP- 3, PYST1) 3, PYST1) Enzyme 3.5 Protein tyrosine phosphatase, PTEN Protein tyrosine phosphatase, PTEN (MMAC1) (MMAC1) Enzyme 3.5 Protein tyrosine phosphatase, PTP4A1 (PRL- Protein tyrosine phosphatase, PTP4A1 (PRL- 1) 1) Enzyme 3.5 Protein tyrosine phosphatase, PTP4A2 (PRL- Protein tyrosine phosphatase, PTP4A2 (PRL- 2) 2) Enzyme 3.5 Protein tyrosine phosphatase, PTP4A3 (PRL- Protein tyrosine phosphatase, PTP4A3 (PRL- 3) 3) Enzyme 3.5 Protein tyrosine phosphatase, PTPN1 Protein tyrosine phosphatase, PTPN1 (PTP1B) (PTP1B) Enzyme 3.5 Protein tyrosine phosphatase, PTPN11 (SHP- Protein tyrosine phosphatase, PTPN11 (SHP- 2) 2) Enzyme 3.5 Protein tyrosine phosphatase, PTPN12 (PTP- Protein tyrosine phosphatase, PTPN12 (PTP- PEST) PEST) Enzyme 3.5 Protein tyrosine phosphatase, PTPN2 Protein tyrosine phosphatase, PTPN2 Enzyme 3.5 (TCPTP) (TCPTP) Protein tyrosine phosphatase, PTPN6 Protein tyrosine phosphatase, PTPN6 (PTP1C, SHP-1) (PTP1C, SHP-1) Enzyme 3.5 Protein tyrosine phosphatase, PTPN9 Protein tyrosine phosphatase, PTPN9 (MEG2) (MEG2) Enzyme 3.5 Protein tyrosine phosphatase, PTPRB Protein tyrosine phosphatase, PTPRB (PTPB) (PTPB) Enzyme 3.5 Protein tyrosine phosphatase, PTPRC Protein tyrosine phosphatase, PTPRC (CD45) (CD45) Enzyme 3.5 Protein tyrosine phosphatase, PTPRF (LAR) Protein tyrosine phosphatase, PTPRF (LAR) Enzyme 3.5 Protein tyrosine phosphatase, PTPRG Protein tyrosine phosphatase, PTPRG (RPTPG) (RPTPG) Enzyme 3.5 Protein tyrosine phosphatase, PTPRJ (DEP1) Protein tyrosine phosphatase, PTPRJ (DEP1) Enzyme 3.5 Protein tyrosine phosphatase, PTPRM Protein tyrosine phosphatase, PTPRM (RPTPM) (RPTPM) Enzyme 3.5

Purinergic P2X receptor Purinergic P2X receptor Ion channel 3.5

Purinergic P2Y receptor Purinergic P2Y receptor Receptor 3.5 Retinoid X Receptor RXR Retinoid X Receptor RXR Receptor 3.5

Ryanodine RyR3 receptor Ryanodine RyR3 receptor Receptor 3.5

Serotonin 5-HT1A receptor Serotonin 5-HT1A receptor Receptor 3.5

Serotonin 5-HT1B receptor Serotonin 5-HT1B receptor Receptor 3.5

Serotonin 5-HT2B receptor Serotonin 5-HT2B receptor Receptor 3.5

Serotonin 5-HT2C receptor Serotonin 5-HT2C receptor Receptor 3.5

Serotonin 5-HT3 receptor Serotonin 5-HT3 receptor Ion channel 3.5

Serotonin 5-HT4 receptor Serotonin 5-HT4 receptor Receptor 3.5

Serotonin 5-HT6 receptor Serotonin 5-HT6 receptor Receptor 3.5

σ1 receptor σ1 receptor Receptor 3.5

σ2 receptor σ2 receptor Receptor 3.5 Sodium channel, site 2 Sodium channel, site 2 Ion channel 3.5

Somatostatin sst1 receptor Somatostatin sst1 receptor Receptor 3.5

Somatostatin sst2 receptor Somatostatin sst2 receptor Receptor 3.5

Somatostatin sst3 receptor Somatostatin sst3 receptor Receptor 3.5 Somatostatin sst4 receptor Somatostatin sst4 receptor Receptor 3.5

Somatostatin sst5 receptor Somatostatin sst5 receptor Receptor 3.5 Sphingomyelinase, acid (A-SMase) Sphingomyelinase, acid (A-SMase) Enzyme 3.5 Sphingomyelinase, neutral (N-SMase) Sphingomyelinase, neutral (N-SMase) Enzyme 3.5 Steroid 5-reductase Steroid 5-reductase Enzyme 3.5

Tachykinin NK1 receptor Tachykinin NK1 receptor Receptor 3.5

Tachykinin NK2 receptor Tachykinin NK2 receptor Receptor 3.5

Tachykinin NK3 receptor Tachykinin NK3 receptor Receptor 3.5 Androgen receptor Androgen receptor Receptor 3.5 Thioredoxin reductase Thioredoxin reductase Enzyme 3.5 Thromboxane synthase Thromboxane synthase Enzyme 3.5 Thyroid hormone receptor Thyroid hormone receptor Receptor 3.5 Thyrotropin releasing hormone (TRH) Thyrotropin releasing hormone (TRH) receptor receptor Receptor 3.5 Transforming Growth Factor- (TGF) Transforming Growth Factor- (TGF) receptor receptor Enzyme 3.5 Transporter, adenosine Transporter, adenosine Transporter 5.7 6.2 Transporter, choline Transporter, choline Transporter 3.5 Transporter, dopamine (DAT) Transporter, dopamine (DAT) Transporter 3.5 Transporter, GABA Transporter, GABA Transporter 3.5 Transporter, glycine Transporter, glycine Transporter 3.5 Transporter, monoamine Transporter, monoamine Transporter 5.7 5.8 Transporter, noradrenaline (NET) Transporter, noradrenaline (NET) Transporter 3.5 Transporter, serotonin (SET) - 5-HT binding Transporter, serotonin (SET) - 5-HT binding site site Transporter 3.5 Transporter, serotonin (SET) - cocaine Transporter, serotonin (SET) - cocaine binding site binding site Transporter 3.5 Tumor necrosis factor (TNF), non-selective Tumor necrosis factor (TNF), non-selective Receptor 3.5 Tyrosine hydroxylase Tyrosine hydroxylase Enzyme 3.5 UDP glucuronosyltransferase, UGT1A1 UDP glucuronosyltransferase, UGT1A1 Enzyme 6.8 UT (Urotensin II) receptor UT (Urotensin II) receptor Receptor 3.5 Vanilloid receptor Vanilloid receptor Ion channel 3.5 Vascular Endothelial Growth Factor (VEGF) Vascular Endothelial Growth Factor (VEGF) Enzyme 3.5 Vasoactive intestinal peptide (VIP1) receptor Vasoactive intestinal peptide (VIP1) receptor Receptor 3.5 Vasopressin V1A receptor Vasopressin V1A receptor Receptor 3.5

Vasopressin V1B receptor Vasopressin V1B receptor Receptor 3.5

Vasopressin V2 receptor Vasopressin V2 receptor Receptor 3.5

Vitamin D3 receptor Vitamin D3 receptor Receptor 3.5 Xanthine oxidase Xanthine oxidase Enzyme 3.5 Supplemental data table 3. Full in vitro kinase profile of R406

Full in vitro profile of R406 at a total of 387 distinct kinases and kinase domains. Kinase naming standards are not uniformly applied, therefore the

KINOMEscan and Millipore assay identifiers are provided for reference to protocol information available on the DiscoveRx

(http://www.discoverx.com/services/drug-discovery-development-services/kinase-profiling/kinomescan) and Merck Millipore

(http://www.millipore.com/life_sciences/flx4/ld_kinases#tab1=3) websites, respectively. Where confidence intervals are provided, these are generated from the global fit of concentration-response replicates. Where no confidence intervals are stated, either a single experiment was performed (duplicate concentration-response curves generated at the same time, on the same plate: n = 1) or the replicate experiments (n = 2 or 3 duplicates, KINOMEscan assays only) were in good agreement and the mean of the individual curve midpoint (Kd) values is presented, thus no confidence intervals were calculated. Kd or IC50

>10,000 nM indicates no activity in the assay. In a small number of instances, there are multiple assays testing interactions at different domains of a single kinase; these share a Kinase gene symbol and have distinct KINOMEscan identifiers.

KINOM Escan confiden Millipore Kinase Target symbol KINOMEscan ce confidence gene KINOMEscan Millipore for x-axis in % inhibition at KINOMEsc KINOMEscan Kd intervals Millipore IC50 intervals symbol identifier identifier Figure 1, 2 10 µM an curve na (nM) (nM)b (nM) (nM)b AAK1 AAK1 AAK1 89 3 290 ABL1- nonphosphorylate ABL1 d ABL1 86 3 1120 760, 1640 960, 3770 ABL1- ABL1 phosphorylated Abl(h) ABL1(p) 87 1 620 1900 960, 3770 2360, ABL2 ABL2 Arg(m) ABL2 77 3 2960 3730 730 570, 930 ACVR1 ACVR1 ACVR1 30 ACVR1B ACVR1B ACVR1B 55 3 2930 2360, 3640 ACVR2A ACVR2A ACVR2A 0 ACVR2B ACVR2B ACVR2B 0 ACVRL1 ACVRL1 ACVRL1 0 ADCK4 ADCK4 ADCK4 100 1 76 AKT1 AKT1 AKT1 0 AKT2 AKT2 AKT2 0 AKT3 AKT3 AKT3 0 ALK ALK ALK(h) ALK 99 1 400 930 420, 2070 ANKK1 ANKK1 ANKK1 95 1 570 Aurora- AURKA AURKA A(h) AURKA 80 1 28 70 50, 90 Aurora- AURKB AURKB B(h) AURKB 100 1 180 30 20, 30 Aurora- AURKC AURKC C(h) AURKC 95 1 110 200 150, 260 AXL AXL Axl(h) AXL 100 1 180 530 330, 840 BLK BLK Blk(h) BLK 100 1 22 70 50, 90 BMP2K BIKE BMP2K 96 1 880 BMPR1A BMPR1A BMPR1A 0 BMPR1B BMPR1B BMPR1B 74 3 760 BMPR2 BMPR2 BMPR2 73 BMX BMX Bmx(h) BMX 89 1 440 30 20, 30 BRAF BRAF BRAF 84 1 260 BRSK1 BRSK1 BRSK1 0 BRSK2 BRSK2 BRSK2 0 BTK BTK BTK(h) BTK 5 1 360 210 160, 270 3050, CABC1 ADCK3 CABC1 84 3 3790 4710 CAMK1 CAMK1 CaMKI(h) CAMK1 97 1 550 1190 860, 1650 CaMKIδ(h CAMK1D CAMK1D ) CAMK1D 98 1 220 410 220, 790 CAMK1G CAMK1G CAMK1G 70 3 840 CAMK2A CAMK2A CAMK2A 62 3 960 CAMK2B CAMK2B CAMK2B 41 CaMKIIδ( CAMK2D CAMK2D h) CAMK2D 49 3 810 690 400, 1180 CaMKIIγ( CAMK2G CAMK2G h) CAMK2G 47 3 840 1000 650, 1530 CAMK4 CAMK4 CAMK4 0 1 >10000 CAMKK1 CAMKK1 CAMKK1 38 CAMKK2 CAMKK2 CAMKK2 9 CASK CASK CASK 47 CDK1/cycl CDC2 inB(h) CDC2 2310 1490, 3570 CDC2L1 CDC2L1 CDC2L1 0 CDC2L2 CDC2L2 CDC2L2 0 CDC2L5 CDC2L5 CDC2L5 0 CDC2L6 CDK11 CDC2L6 0 CDC42BPA MRCKA CDC42BPA 0 CDC42BPB MRCKB CDC42BPB 0 CDC42BPG DMPK2 CDC42BPG 26 CDK2 CDK2 CDK2 38 1 >10000 CDK3 CDK3 CDK3 46 1 >10000 CDK4 CDK4-cyclinD1 CDK4(D1) 0 CDK4 CDK4-cyclinD3 CDK4(D3) 6 CDK5 CDK5 CDK5 0 1 >10000 CDK7 CDK7 CDK7 0 1 >10000 CDK8 CDK8 CDK8 0 CDK9 CDK9 CDK9 0 1500, CDKL1 CDKL1 CDKL1 84 3 2740 5010 CDKL2 CDKL2 CDKL2 58 CDKL3 CDKL3 CDKL3 0 CDKL5 CDKL5 CDKL5 0 2330, CHEK1 CHEK1 CHEK1 62 3 9060 35270 1040, CHEK2 CHEK2 CHK2(h) CHEK2 45 3 1270 1540 670 520, 870 CHUK IKK-alpha IKKα(h) CHUK 98 2 15 >10000 CIT CIT CIT 99 1 930 CLK1 CLK1 CLK1 92 1 760 CLK2 CLK2 CLK2(h) CLK2 84 1 620 650 520, 820 CLK3 CLK3 CLK3 78 3 3790 1340, 10720 CLK4 CLK4 CLK4 85 1 260 CSF1R CSF1R Fms(h) CSF1R 100 1 41 20 20, 30 CSK CSK CSK(h) CSK 95 1 480 1730 710, 4210 CSNK1A1 CSNK1A1 CSNK1A1 24 CSNK1A1L CSNK1A1L CSNK1A1L 0 CSNK1D CSNK1D CSNK1D 0 CSNK1E CSNK1E CSNK1E 0 CSNK1G1 CSNK1G1 CSNK1G1 0 CSNK1G2 CSNK1G2 CSNK1G2 0 CSNK1G3 CSNK1G3 CSNK1G3 0 CSNK2A1 CSNK2A1 CK2(h) CSNK2A1 100 1 58 630 460, 850 CSNK2A2 CSNK2A2 CK2α2(h) CSNK2A2 100 1 14 380 280, 520 1090, DAPK1 DAPK1 DAPK1(h) DAPK1 97 1 100 3490 11050 DAPK2 DAPK2 DAPK2(h) DAPK2 95 1 160 1360 900, 2060 DAPK3 DAPK3 ZIPK(h) DAPK3 96 1 18 450 300, 690 DCLK1 DCAMKL1 DCLK1 59 DCLK2 DCAMKL2 DCLK2 36 DCLK3 DCAMKL3 DCLK3 96 3 1130 780, 1650 DDR1 DDR1 DDR1 100 1 15 DDR2 DDR2 DDR2(h) DDR2 86 1 20 240 180, 320 DMPK DMPK DMPK 89 1 >10000 DSTYK RIPK5 DSTYK 100 1 >10000 DYRK1A DYRK1A DYRK1A 91 1 320 DYRK1B DYRK1B DYRK1B 78 DYRK2 DYRK2 DYRK2(h) DYRK2 91 1 760 >10000 EGFR EGFR EGFR(h) EGFR 98 1 170 940 650, 1340 EIF2AK1 EIF2AK1 EIF2AK1 38 EIF2AK2 PRKR EIF2AK2 86 1 88 GCN2(Kin.Dom. EIF2AK4 2,S808G) EIF2AK4 100 1 3.7 EPHA1 EPHA1 EphA1(h) EPHA1 98 1 160 80 70, 100 EPHA2 EPHA2 EphA2(h) EPHA2 90 3 330 260, 410 300 250, 360 EPHA3 EPHA3 EphA3(h) EPHA3 90 1 960 1100 460, 2660 EPHA4 EPHA4 EphA4(h) EPHA4 69 3 540 920 620, 1350 EPHA5 EPHA5 EPHA5 64 3 2650 2180, 3230 EPHA6 EPHA6 EPHA6 82 1 420 EPHA7 EPHA7 EphA7(h) EPHA7 100 3 54 210 170, 270 1160, EPHA8 EPHA8 EPHA8 58 3 7090 43470 EPHB1 EPHB1 EphB1(h) EPHB1 96 3 1180 920, 1520 80 60, 100 2810, EPHB2 EPHB2 EPHB2 50 3 3830 5220 EPHB3 EPHB3 EPHB3 0 1320, EPHB4 EPHB4 EPHB4 50 3 1640 2040 EPHB6 EPHB6 EPHB6 93 1 610 ERBB2 ERBB2 ERBB2 70 ERBB3 ERBB3 ERBB3 83 1 >10000 ERBB4 ERBB4 ErbB4(h) ERBB4 89 1 440 580 310, 1080 1170, ERN1 ERN1 ERN1 70 3 2030 3520 FER FER Fer(h) FER 78 3 190 620 380, 1020 FES FES Fes(h) FES 98 3 550 410, 740 190 120, 310 FGFR1 FGFR1 FGFR1(h) FGFR1 100 1 49 30 20, 40 FGFR2 FGFR2 FGFR2(h) FGFR2 96 1 110 60 50, 70 FGFR3 FGFR3 FGFR3(h) FGFR3 99 1 74 40 30, 50 FGFR4 FGFR4 FGFR4(h) FGFR4 97 1 180 >10000 FGR FGR Fgr(h) FGR 96 1 70 120 90, 160 FLT1 FLT1 Flt1(h) FLT1 100 1 24 4 4, 5 FLT3 FLT3 Flt3(h) FLT3 100 3 0.97 3 2, 3 FLT4 FLT4 Flt4(h) FLT4 100 3 15 4 3, 4 FRAP1 MTOR FRAP1 1 FRK FRK PTK5(h) FRK 96 1 190 50 40, 70 FYN FYN Fyn(h) FYN 94 1 78 50 40, 60 GAK GAK GAK 95 1 240 GRK1 GRK1 GRK1 57 3 >10000 GRK4 GRK4 GRK4 0 3 2220 970, 5060 GRK7 GRK7 GRK7 49 3 >10000 1630, GSK3A GSK3A GSK3A 60 3 2900 5150 GSK3B GSK3B GSK3β(h) GSK3B 58 3 170 550 360, 830 HCK HCK Hck(h) HCK 98 1 180 90 70, 120 1790, HIPK1 HIPK1 HIPK1(h) HIPK1 65 3 3370 6350 >10000 HIPK2 HIPK2 HIPK2 59 HIPK3 HIPK3 HIPK3 64 HIPK4 HIPK4 HIPK4 80 3 >10000 HUNK HUNK HUNK 0 ICK ICK ICK 23 IGF1R IGF1R IGF1R 81 3 >10000 IKBKB IKK-beta IKKβ(h) IKBKB 99 1 140 210 130, 320 IKBKE IKK-epsilon IKBKE 99 1 42 INSR INSR IR(h) INSR 65 1 790 450 210, 970 INSRR INSRR INSRR 53 IRAK1 IRAK1 IRAK1(h) IRAK1 100 1 16 60 50, 80 IRAK3 IRAK3 IRAK3 96 1 5 IRAK4 IRAK4 IRAK4(h) IRAK4 96 1 400 430 340, 540 ITK ITK Itk(h) ITK 57 3 750 720 580, 890 JAK1(JH1domain JAK1 -catalytic) JAK1 100 1 38 JAK1(JH2domain 1430, JAK1 -pseudokinase) JAK1(JH2) 98 3 1980 2740 JAK2(JH1domain JAK2 -catalytic) JAK2(h) JAK2 99 1 0.78 7 6, 8 JAK3(JH1domain JAK3 -catalytic) JAK3(h) JAK3 100 1 10 70 60, 80 KDR VEGFR2 KDR(h) KDR 99 3 76 30 20, 40 KIAA0999 QSK KIAA0999 35 KIT KIT cKit(h) KIT 100 3 8.1 580 420, 790 LATS1 LATS1 LATS1 39 LATS2 LATS2 LATS2 100 1 >10000 LCK LCK Lck(h) LCK 100 1 30 30 20, 40 2770, LIMK1 LIMK1 LIMK1 85 3 4710 8030 1520, LIMK2 LIMK2 LIMK2 78 3 1950 2490 LRRK2 LRRK2 LRRK2 97 1 300 LTK LTK LTK 97 1 370 LYN LYN Lyn(h) LYN 98 1 81 40 30, 40 MAK MAK MAK 0 MAL13P1.2 PFPK5(P.falcipar 79 um) MAL13P1.279 0 MAP2K1 MEK1 MEK1(h) MAP2K1 94 1 280 >10000 1530, MAP2K2 MEK2 MAP2K2 87 3 1890 2340 MAP2K3 MEK3 MAP2K3 94 1 140 MAP2K4 MEK4 MKK4(m) MAP2K4 85 1 230 >10000 MAP2K5 MEK5 MAP2K5 100 1 26 MAP2K6 MEK6 MKK6(h) MAP2K6 91 1 650 1110 610, 2040 MAP2K7 MKK7 MAP2K7 8 1 >10000 MAP3K1 MAP3K1 MAP3K1 37 MAP3K10 MLK2 MAP3K10 100 1 3 MAP3K11 MLK3 MAP3K11 100 1 12 MAP3K12 DLK MAP3K12 32 MAP3K13 LZK MAP3K13 22 MAP3K15 MAP3K15 MAP3K15 51 MAP3K2 MAP3K2 MAP3K2 99 1 170 MAP3K3 MAP3K3 MAP3K3 98 1 260 1070, MAP3K4 MAP3K4 MAP3K4 71 3 1710 2730 MAP3K5 ASK1 MAP3K5 0 MAP3K6 ASK2 MAP3K6 32 MAP3K7 TAK1 TAK1(h) MAP3K7 0 1 100 470 320, 670 MAP3K9 MLK1 MLK1(h) MAP3K9 99 1 10 5 4, 6 2080, MAP4K1 HPK1 MAP4K1 70 3 2950 4190 MAP4K2 MAP4K2 GCK(h) MAP4K2 85 3 990 660, 1490 1190 610, 2330 1800, MAP4K3 MAP4K3 MAP4K3 52 3 3690 7600 MAP4K4 MAP4K4 MAP4K4 33 MAP4K5 MAP4K5 MAP4K5 16 MAPK1 ERK2 MAPK1 0 MAPK10 JNK3 JNK3(h) MAPK10 100 1 16 40 30, 60 MAPK11 p38-beta MAPK11 0 MAPK12 p38-gamma SAPK3(h) MAPK12 99 2 420 300, 570 >10000 MAPK13 p38-delta SAPK4(h) MAPK13 100 1 460 220 150, 320 MAPK14 p38-alpha MAPK14 5 MAPK15 ERK8 MAPK15 89 1 300 MAPK3 ERK1 MAPK3 0 MAPK4 ERK4 MAPK4 14 MAPK6 ERK3 MAPK6 43 1 >10000 MAPK7 ERK5 MAPK7 19 MAPK8 JNK1 JNK1α1(h) MAPK8 100 1 46 >10000 MAPK9 JNK2 JNK2α2(h) MAPK9 100 1 36 650 440, 940 MAPKAPK 2 MAPKAPK2 MAPKAPK2 0 MAPKAPK 5 MAPKAPK5 MAPKAPK5 19 MARK1(h MARK1 MARK1 ) MARK1 94 3 170 130, 220 600 440, 800 PAR- MARK2 MARK2 1Bα(h) MARK2 88 1 250 450 300, 640 MARK3 MARK3 MARK3 63 3 350 MARK4 MARK4 MARK4 99 1 100 MAST1 MAST1 MAST1 39 MATK CTK MATK 70 MELK MELK MELK 71 MERTK MERTK Mer(h) MERTK 100 1 320 260 170, 400 MET MET Met(h) MET 97 1 520 270 160, 470 MGC42105 NIM1 MGC42105 84 3 950 780, 1160 MINK1 MINK MINK1 72 1650, MKNK1 MKNK1 MKNK1 9 3 4180 10620 MKNK2 MKNK2 Mnk2(h) MKNK2 100 1 72 300 250, 360 MST1R MST1R Ron(h) MST1R 100 1 310 100 80, 130 MST4 MST4 MST4 51 MUSK MUSK MuSK(h) MUSK 99 1 160 210 150, 290 MYLK MYLK MYLK 54 MYLK2 MYLK2 MYLK2 58 MYLK3 MLCK MYLK3 30 MYLK4 MYLK4 MYLK4 84 1 620 MYO3A MYO3A MYO3A 10 MYO3B MYO3B MYO3B 0 NEK1 NEK1 NEK1 96 1 330 NEK11 NEK11 NEK11(h) NEK11 100 1 640 1550 1130, 2120 NEK2 NEK2 NEK2(h) NEK2 83 3 1100 870, 1400 910 460, 1790 NEK3 NEK3 NEK3(h) NEK3 99 1 200 430 290, 530 1040, NEK4 NEK4 NEK4 91 3 1480 2120 NEK5 NEK5 NEK5 98 3 8.5 NEK6 NEK6 NEK6(h) NEK6 100 1 3.6 >10000 NEK7 NEK7 NEK7(h) NEK7 100 1 31 >10000 NEK9 NEK9 NEK9 97 1 14 NLK NLK NLK 0 NTRK1 TRKA TrkA(h) NTRK1 100 3 90 30 20, 30 NTRK2 TRKB TrkB(h) NTRK2 95 3 100 70, 150 310 170, 580 NTRK3 TRKC NTRK3 96 1 63 NUAK1 ARK5 ARK5(h) NUAK1 93 1 96 60 40, 70 NUAK2 SNARK NUAK2 97 1 160 OXSR1 OSR1 OXSR1 71 PAK1 PAK1 PAK1 91 3 220 200, 240 PAK2 PAK2 PAK2(h) PAK2 64 1 540 500 310, 820 PAK3 PAK3 PAK3 99 1 260 PAK4 PAK4 PAK4(h) PAK4 97 1 140 140 110, 160 PAK6 PAK6 PAK6(h) PAK6 96 2 160 440 300, 630 PAK7 PAK7 PAK5(h) PAK7 100 1 56 160 90, 270 PCTK1 PCTK1 PCTK1 54 PCTK2 PCTK2 PCTK2 60 1 920 PCTK3 PCTK3 PCTK3 0 PDGFRα(h PDGFRA PDGFRA ) PDGFRA 100 1 36 190 120, 300 PDGFRβ(h PDGFRB PDGFRB ) PDGFRB 100 1 2.6 200 130, 320 PDPK1 PDPK1 PDK1(h) PDPK1 78 1 110 120 100, 140 PFCDPK1(P.falci PFB0815w parum) PFB0815w 74 PFTK1 PFTK1 PFTK1 0 PFTK2 PFTAIRE2 PFTK2 2 PHKG1 PHKG1 PHKG1 29 PHKG2 PHKG2 PhKγ2(h) PHKG2 82 1 980 >10000 PI4KB PIK4CB PI4KB 66 3 2320 990, 5450 PIK3C2B PIK3C2B PIK3C2B 41 PIK3C2G PIK3C2G PIK3C2G 48 PIK3CA PIK3CA PIK3CA 98 1 >10000 PIK3CD PIK3CD PIK3CD 62 PIK3CG PIK3CG PIK3CG 26 PIM1 PIM1 PIM1 24 PIM2 PIM2 PIM2 0 PIM3 PIM3 PIM3 38 1920, PIP4K2B PIP5K2B PIP4K2B 63 3 2800 4080 PIP4K2C PIP5K2C PIP4K2C 100 3 200 100, 410 PIP5K1a(h PIP5K1A PIP5K1A ) PIP5K1A 100 1 660 >10000 PIP5K1g(h PIP5K1C PIP5K1C ) PIP5K1C 100 3 240 >10000 PKMYT1 PKMYT1 PKMYT1 30 PKN1 PKN1 PKN1 96 1 500 1200, PKN2 PKN2 PKN2 70 3 1730 2500 PKNB(M.tubercul PKNB osis) pknB 46 1 24 PLK1 PLK1 Plk1(h) PLK1 87 1 140 910 550, 1530 PLK2 PLK2 Snk(h) PLK2 100 3 49 130 110, 160 PLK3 PLK3 Plk3(h) PLK3 100 1 9.8 50 40, 60 PLK4 PLK4 PLK4 100 1 5 AMPKα1( PRKAA1 AMPK-alpha1 h) PRKAA1 74 3 370 280, 490 960 620, 1480 AMPKα2( PRKAA2 AMPK-alpha2 h) PRKAA2 76 3 460 660 450, 970 PRKACA PKAC-alpha PKA(h) PRKACA 88 3 490 410, 590 700 320, 1540 PRKACB PKAC-beta PRKACB 92 1 280 PRKCB PKCβII(h) PRKCB >10000 PRKCD PRKCD PRKCD 17 PRKCE PRKCE PRKCE 73 PRKCG PKCγ(h) PRKCG 400 250, 660 PRKCH PRKCH PRKCH 0 PRKCI PRKCI PRKCI 35 PRKCQ PRKCQ PRKCQ 30 PRKD1 PRKD1 PRKD1 17 PRKD2 PRKD2 PRKD2 0 PRKD3 PRKD3 PRKD3 0 PRKG1 PRKG1 PRKG1 0 PRKG2 PRKG2 PRKG2 77 PRKX PRKX PRKX 0 PRPF4B PRP4 PRPF4B 26 PTK2 FAK FAK(h) PTK2 98 1 550 310 220, 450 PTK2B PYK2 Pyk2(h) PTK2B 99 1 41 130 90, 180 PTK6 BRK BRK(h) PTK6 82 1 880 460 300, 700 RAF1 RAF1 c-RAF(h) RAF1 69 3 50 30, 70 220 180, 270 RET RET Ret(h) RET 100 1 5.6 10 10, 10 RIOK1 RIOK1 RIOK1 77 1 190 RIOK2 RIOK2 RIOK2 83 1 970 RIOK3 RIOK3 RIOK3 87 1 160 RIPK1 RIPK1 RIPK1 4 RIPK2 RIPK2 RIPK2(h) RIPK2 85 1 1000 790 460, 1350 RIPK4 RIPK4 RIPK4 96 1 340 ROCK- ROCK1 ROCK1 I(h) ROCK1 86 1 720 >10000 ROCK- ROCK2 ROCK2 II(h) ROCK2 86 1 660 830 460, 1470 ROS1 ROS1 Ros(h) ROS1 100 1 22 940 500, 1770 RSK1(Kin.Dom.1 1090, RPS6KA1 -N-terminal) Rsk1(h) RPS6KA1(N) 97 3 1480 2010 740 520, 1060 RSK1(Kin.Dom.2 RPS6KA1 -C-terminal) RPS6KA1(C) 27 3 >10000 RSK3(Kin.Dom.1 RPS6KA2 -N-terminal) RPS6KA2(N) 5 1 >10000 RSK3(Kin.Dom.2 RPS6KA2 -C-terminal) RPS6KA2(C) 0 RSK2(Kin.Dom.1 RPS6KA3 -N-terminal) Rsk2(h) RPS6KA3 88 1 770 520, 1160 330 180, 600 RPS6KA4 RPS6KA4(Kin.D RPS6KA4(N) 0 1 >10000 om.1-N-terminal) RPS6KA4(Kin.D RPS6KA4 om.2-C-terminal) MSK2(h) RPS6KA4(C) 100 3 120 90, 150 >10000 RPS6KA5(Kin.D RPS6KA5 om.1-N-terminal) RPS6KA5(N) 0 RPS6KA5(Kin.D RPS6KA5 om.2-C-terminal) MSK1(h) RPS6KA5(C) 95 1 140 >10000 RSK4(Kin.Dom.1 RPS6KA6 -N-terminal) RPS6KA6(N) 70 RSK4(Kin.Dom.2 RPS6KA6 -C-terminal) RPS6KA6(C) 66 RPS6KB1 S6K1 RPS6KB1 81 1 >10000 SBK1 SBK1 SBK1 90 1 630 SgK110 SgK110 SgK110 91 1 220 SGK3 SGK3 SGK3 52 SIK1 SIK SIK(h) SIK1 100 1 40 70 50, 100 SIK2 SIK2 SIK2 60 1 100 SLK SLK SLK 100 1 53 SNRK SNRK SNRK 7 SRC SRC cSRC(h) SRC 100 1 16 20 10, 20 SRMS SRMS SRMS 86 3 1270 780, 2060 SRPK1 SRPK1 SRPK1(h) SRPK1 87 1 340 >10000 SRPK2 SRPK2 SRPK2 0 1480, SRPK3 SRPK3 MSSK1(h) SRPK3 12 3 3530 8410 >10000 STK10 LOK LOK(h) STK10 100 3 110 90, 130 140 100, 210 STK11 LKB1 STK11 0 STK16 STK16 STK16 100 3 0.6 0.5, 0.8 STK17A DRAK1 DRAK1(h) STK17A 100 1 86 190 140, 260 STK17B DRAK2 STK17B 96 3 15 STK24 MST3 STK24 21 STK25 YSK1 STK25 80 3 >10000 STK3 MST2 STK3 74 STK32A YANK1 STK32A 72 3 1210 760, 1930 STK32B YANK2 STK32B 0 STK32C YANK3 STK32C 0 STK33 STK33 STK33(h) STK33 85 1 310 470 270, 810 3360, STK35 STK35 STK35 71 3 5510 9040 STK36 STK36 STK36 95 1 440 STK38 NDR1 STK38 65 STK38L NDR2 STK38L 48 STK39 STK39 STK39 65 STK4 MST1 STK4 0 SYK SYK Syk(h) SYK 98 1 12 50 40, 60 TAOK1 TAOK1 TAO1(h) TAOK1 92 1 1100 980 620, 1540 TAOK2 TAOK2 TAO2(h) TAOK2 88 3 1120 720, 1740 140 90, 220 TAOK3 TAOK3 TAOK3 66 TBK1 TBK1 TBK1(h) TBK1 100 1 22 20 10, 20 Tec(h) TEC TEC activated TEC 82 3 760 100 70, 160 TEK TIE2 Tie2 (h) TEK 94 3 520 200 140, 290 1400, TESK1 TESK1 TESK1 83 3 1860 2470 2570, TGFBR1 TGFBR1 TGFBR1 45 3 4360 7400 TGFBR2 TGFBR2 TGFBR2 75 1 820 TIE1 TIE1 TIE1 94 3 100 TLK1 TLK1 TLK1 81 3 310 TLK2 TLK2 TLK2(h) TLK2 86 3 240 1230 950, 1590 1670, TNIK TNIK TNIK 56 3 2230 2990 TNK1 TNK1 TNK1 100 3 24 TNK2 TNK2 ACK1(h) TNK2 99 3 120 20 10, 20 TNNI3K TNNI3K TNNI3K 94 3 520 TRPM6 TRPM6 TRPM6 0 TSSK1B TSSK1B TSSK1(h) TSSK1B 97 3 100 60, 160 50 30, 70 1010, TTK TTK TTK 81 3 1290 1630 TXK TXK Txk(h) TXK 90 3 230 210, 260 100 50, 170 TYK2(JH1domai TYK2 n-catalytic) TYK2 90 3 16 TYK2(JH2domai 3040, TYK2 n-pseudokinase) TYK2(JH2) 44 3 5290 9190 3890, TYRO3 TYRO3 TYRO3 61 3 11580 34490 ULK1 ULK1 ULK1 100 3 14 ULK2 ULK2 ULK2(h) ULK2 100 3 31 60 40, 80 ULK3 ULK3 ULK3(h) ULK3 100 3 40 30, 50 20 20, 20 VRK2 VRK2 VRK2 0 1700, WEE1 WEE1 WEE1 81 3 2120 2640 WEE2 WEE2 WEE2 95 3 >10000 YES1 YES Yes(h) YES1 98 3 120 40 30, 60 YSK4 YSK4 YSK4 100 3 22 ZAK ZAK ZAK 44 ZAP70 ZAP70 ZAP-70(h) ZAP70 100 3 34 1300 830, 2040 a Number of independent replicates (duplicates) performed on separate days using separate samples. b 95% lower and upper confidence intervals from the global fit of concentration-response curve replicates.