Pharmacological Characterization of AZD5069, a Slowly Reversible CXCR2 Antagonist Nicholls DJ, Wiley K, Dainty I, MacIntosh F, Phillips C, Gaw A, Kärrman Mårdh C. JPET #221358

Pharmacological Characterization of AZD5069, a Slowly Reversible CXCR2 Antagonist

David Nicholls, Katherine Wiley, Ian Dainty, Fraser MacIntosh, Caroline Phillips, Alasdair Gaw, Carina Kärrman Mårdh. The Journal of Pharmacology and Experimental Therapeutics

Table S1. Kinase panel screening performed at Dundee University

ATP Inhibition Target (μM) (%) AMP-activated Kinase 50 19 Cyclin Dependent Kinase 2:Cyclin A 20 18 Checkpoint Kinase I 20 10 Casein Kinase 1 20 -6 Casein Kinase 2 5 4 Lymphocyte-Specific Protein Tyrosine Kinase 50 -5 Mitogen Activated Protein Kinase 1 50 13 MAPK Activated Protein Kinase-1a 50 -3 MAPK Activated Protein Kinase-2 20 13 Mitogen Activated Protein Kinase Kinase 1 5 21 Protein Kinase, cAMP-dependent, catalytic, alpha 5 -71 v-akt murine thymoma viral oncogene homolog 1 5 19 Protein Kinase C alpha 20 9 p38 Regulated Activated Kinase 20 7 Dual Specificity Tyrosine Phosphorylation Regulated Kinase 1A 50 15 Glycogen Synthase Kinase-3 beta 5 12 Mitogen Activated Protein Kinase 8 20 9 Mitogen and Stress Activated Protein Kinase 1 20 8 P70S6 Kinase 20 22 Phospho B Kinase 20 4 3-Phosphoinositide Dependent Protein Kinase-1 20 10 Rho Associated, Coiled Coil Containing Protein Kinase 2 20 2 Stress Activated Protein Kinase 2a 50 4 Stress Activated Protein Kinase 2b 20 17 Serum and Glucocorticoid Regulated Kinase? 20 11

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Pharmacological Characterization of AZD5069, a Slowly Reversible CXCR2 Antagonist Nicholls DJ, Wiley K, Dainty I, MacIntosh F, Phillips C, Gaw A, Kärrman Mårdh C. JPET #221358

Table S2. Secondary pharmacology screening performed at MDS Pharma

Conc Inhibition Cat# Target (μM) (%) 138000 15 Lipoxygenase 10 -7 136010 5 Lipoxygenase 10 -4 271110 5-Hydroxytryptamine 1A 10 -15 271200 5-Hydroxytryptamine Receptor 1B 10 -5 271650 5-Hydroxytryptamine Receptor 2A 10 2 271700 5-Hydroxytryptamine Receptor 2B 10 5 271800 5-Hydroxytryptamine Receptor 2C 10 -7 271910 5-Hydroxytryptamine Receptor 3A 10 -12 272000 5-Hydroxytryptamine Receptor 4 10 -4 272100 5-Hydroxytryptamine Receptor 5A 10 14 272200 5-Hydroxytryptamine Receptor 5B 10 -1 104010 Acetylcholinesterase 10 9 200510 A1 10 -4 200610 Adenosine Receptor A2a 10 -4 200720 Adenosine Receptor A3 10 5 202000 Adenosine Transporter 10 -2 203100 Alpha 1A 10 21 203200 Adrenergic Receptor Alpha 1B 10 -5 203400 Adrenergic Receptor Alpha 1D 10 10 203620 Adrenergic Receptor Alpha 2A 10 7 203710 Adrenergic Receptor Alpha 2B 10 -7 203800 Adrenergic Receptor Alpha 2C 10 2 204010 Adrenergic Receptor Beta 1 10 1 204110 Adrenergic Receptor Beta 2 10 -4 204200 Adrenergic Receptor Beta 3 10 6 107000 Aldose Reductase 10 75 232600 AMPA/Kainate Receptor 10 12 285010 Androgen Receptor 10 6 107300 Angiotensin Converting Enzyme 10 0 210020 Angiotensin II Receptor AT1 10 3 210110 Angiotensin II Receptor AT2 10 5 265600 ATP-sensitive Potasium Channel 10 3 211600 BRS-1 30 3 211700 Bombesin Receptor BRS-2 30 2 212510 1 10 8 212610 Bradykinin Receptor 2 30 -5 213610 30 8 216000 Calcium channel 30 4 214600 Calcium channel 10 7 214510 Calcium channel 10 3 215000 Calcium channel 10 -5

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Pharmacological Characterization of AZD5069, a Slowly Reversible CXCR2 Antagonist Nicholls DJ, Wiley K, Dainty I, MacIntosh F, Phillips C, Gaw A, Kärrman Mårdh C. JPET #221358

108010 Calpain 10 5 217020 1 30 -3 217100 Cannabinoid Receptor 2 30 -9 112000 Carbonic Anhydrase 10 31 112020 Carbonic Anhydrase 10 2 163000 Caspase 1 10 -1 163200 Caspase 3 10 -1 163280 Caspase 5 10 -14 163300 Caspase 6 10 0 112250 Cathepsin B 10 -19 112510 Cathepsin G 10 4 217500 CCR1 30 4 217550 CCR2 30 73 217650 CCR4 30 -2 217700 CCR5 30 58 214010 CGRP Receptor 10 -7 218010 Cholecystokinin A Receptor 30 -1 218120 Cholecystokinin B Receptor 30 13 219000 Choline Transporter 10 7 113400 Chymotrypsin 10 4 219150 Corticotropin releasing 1 10 3 219500 D1 10 -2 219600 10 6 219700 Dopamine Receptor D2 10 -13 219800 10 -1 219900 10 1 220000 Dopamine Receptor D4 10 3 220100 Dopamine Receptor D4 10 -1 220200 10 -3 220320 Dopamine Transporter 10 2 163950 Endothelin Converting Enzyme 1 10 20 224010 A 10 7 224110 Endothelin Receptor B 30 9 143000 eNOS 10 5 170010 Epidermal Growth Factor Receptor 10 11 171790 Fibroblast Growth Factor Receptor 1 10 -4 172010 FYN oncogene related to SRC 10 1 226500 GABA Receptor 10 7 226600 GABA Receptor 10 8 228700 GABAB Receptor 10 7 228600 GABAB Receptor 10 -3 231510 1 30 2 231600 30 3 239300 Ghrelin Receptor 30 10 231700 Glucagon Like Peptide 1 Receptor 30 2

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Pharmacological Characterization of AZD5069, a Slowly Reversible CXCR2 Antagonist Nicholls DJ, Wiley K, Dainty I, MacIntosh F, Phillips C, Gaw A, Kärrman Mårdh C. JPET #221358

232010 Glucocorticoid Receptor 10 4 239000 Glycine receptor 10 -9 239100 Glycine 10 27 239610 H1 10 1 239710 Histamine Receptor H2 10 -3 239810 Histamine Receptor H3 10 14 239900 Histamine Receptor H4 10 -20 241000 Imidazoline I2 10 3 144000 iNOS 10 -7 175000 Insulin Receptor 10 16 243000 Insulin Receptor 10 7 232700 Kainate GLU Receptor 10 -6 286900 Kinase Insert Domain Receptor 30 0 250400 Leptin Receptor 10 10 250510 10 -9 250460 CysT1 10 8 115400 Matrix Metalloproteinase 14 10 -7 114210 Matrix Metalloproteinase 2 10 -5 251100 1 30 -7 251300 Melanocortin Receptor 3 30 11 251350 Melanocortin Receptor 4 30 3 251400 Melanocortin Receptor 5 30 9 251600 1 30 6 251500 Melatonin Receptor 10 2 175030 Met proto-oncogene (hepatocyte growth factor receptor) 10 7 176700 Mitogen-Activated Protein Kinase 12 10 6 140010 Monoamine Oxidase A 10 0 252200 30 -1 252600 Muscarinic 1 10 -9 252700 Muscarinic Acetylcholine Receptor 2 10 -6 252800 Muscarinic Acetylcholine Receptor 3 10 -7 252900 Muscarinic Acetylcholine Receptor 4 10 2 253000 Muscarinic Acetylcholine Receptor 5 10 -2 254500 Muscarinic Acetylcholine Receptor 10 4 121000 Myeloperoxidase 10 16 107700 Na+/K+ transporting ATPase 10 4 211000 Natriuretic Peptide Receptor 30 7 255510 Neurokinin Receptor 1 10 6 255600 Neurokinin Receptor 2 10 0 255710 Neurokinin Receptor 3 10 11 257010 Y Receptor 1 30 5 257110 2 10 -5 258010 1 10 3 258590 Nicotinic Acetylcholine Receptor 10 -9 232810 NMDA Receptor 10 22

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Pharmacological Characterization of AZD5069, a Slowly Reversible CXCR2 Antagonist Nicholls DJ, Wiley K, Dainty I, MacIntosh F, Phillips C, Gaw A, Kärrman Mårdh C. JPET #221358

233000 NMDA Receptor 10 -2 234000 NMDA Receptor 10 5 232910 NMDA Receptor 10 -13 142000 nNOS 10 0 260600 OPRL1 10 3 204410 Norepinephrine Transporter 10 -1 226010 Oestrogen Receptor Alpha 10 7 226050 Oestrogen Receptor Beta 10 -5 260110 Delta 1 10 -4 260210 Opioid Receptor Kappa 1 10 -9 260410 Opioid Receptor Mu 1 10 -8 160000 Pancreatic Phospholipase A2 10 8 152000 Phosphodiesterase 3 10 3 270000 Phosphodiesterase 4 10 5 154000 Phosphodiesterase 4 10 7 156000 Phosphodiesterase 5 10 13 156100 Phosphodiesterase 6 10 5 265010 Platelet Activating Factor Receptor 30 -1 265200 Platelet Derived Growth Factor Receptor 30 -1 265500 Potassium Channel 30 -18 265800 Potassium Channel 10 -2 268020 Progesterone receptor 10 1 268410 EP4 10 5 116020 Prostaglandin-endoperoxide synthase 1 10 4 118010 Prostaglandin-endoperoxide synthase 2 10 10 190010 Protein Tyrosine Phosphatase 10 10 268810 Purine -coupled Receptor 10 13 268700 Purine receptor 10 3 164600 Renin 10 9 269500 Retinoid X Receptor 10 5 171520 Rho Associated 10 -3 274020 Serotonin Transporter 10 4 274030 Serotonin Transporter 10 -3 278110 Sigma1 10 2 278200 Sigma2 10 3 279510 Sodium Channel 10 4 282510 1 30 3 282700 10 -2 282530 30 3 282900 10 -3 283000 10 10 165000 Thrombin 10 2 285510 A2 10 -8 286000 Thyrotropin Releasing Hormone Receptor 1 10 9 286810 Transient Receptor Potential Cation Channel V 10 -6

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Pharmacological Characterization of AZD5069, a Slowly Reversible CXCR2 Antagonist Nicholls DJ, Wiley K, Dainty I, MacIntosh F, Phillips C, Gaw A, Kärrman Mårdh C. JPET #221358

226700 Translocator Protein (18kDa) 10 18 226400 Undefined 10 13 171800 Vascular Endothelial Growth Factor Receptor 1 10 12 287520 Receptor 1A 10 0 287560 1B 10 6 287610 10 9 252010 Vesicular Monoamine Transporter 10 34 287010 VPAC Receptor 1 30 -2 176800 v-src sarcoma (Schmidt-Ruppin A-2) viral oncogene homolog 10 -1 175080 v-yes-1 Yamaguchi Sarcoma Viral Oncogene Homolog 1 10 -1

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