Supplementary information for Exploration of 3,4-unsubstituted coumarins as thioredoxin reductase 1 inhibitor for cancer therapy A.Nikitjuka1,*, M. Ozola1, L. Jackevica1, R. Bobrovs1, R. Žalubovskis1,2,* 1Latvian Institute of Organic Synthesis, Aizkraukles 21, LV-1006, Riga, Latvia 2Institute of Technology of Organic Chemistry, Faculty of Materials Science and Applied Chemistry, Riga Technical University, P. Valdena iela 3, LV-1048 Riga, Latvia anna@osi.lv; raivis@osi.lv Table of Contents Synthesis of 6-amino-3-methyl-2H-chromen-2-one (11) .....................................................................3 Synthesis of 3-amino-2H-chromen-2-one (12) .....................................................................................3 Reaction of inhibitor 7b with methyl 3-mercaptopropanoate in physiological conditions ...................4 Dose- response curve for compound 7b, TrxR1 IC50............................................................................5 Dose- response curve for compound 7b on different cell lines (representative examples) ...................5 6-(4-(Trifluoromethyl)phenyl)-2H-chromen-2-one (7b) 1H, 13C, 18F, IR, HRMS ................................6 6-(4-Nitrophenyl)-2H-chromen-2-one (8b) 1H, 13C, IR, HRMS...........................................................9 6-Cyclopropyl-2H-chromen-2-one (10b) 1H, 13C, IR, HRMS ............................................................11 5-Nitro-2H-chromen-2-one (3a) 1H and 13C .......................................................................................13 8-Nitro-2H-chromen-2-one (3d) 1H and 13C .......................................................................................14 5-Amino-2H-chromen-2-one (4a) 1H and 13C.....................................................................................15 6-Amino-2H-chromen-2-one (4b) 1H and 13C ....................................................................................16 8-Amino-2H-chromen-2-one (4d) 1H and 13C ....................................................................................17 5-Phenyl-2H-chromen-2-one (5a) 1H and 13C.....................................................................................18 6-Phenyl-2H-chromen-2-one (5b) 1H and 13C ....................................................................................19 7-Phenyl-2H-chromen-2-one (5c) 1H and 13C.....................................................................................20 8-Phenyl-2H-chromen-2-one (5d) 1H and 13C ....................................................................................21 5-(Pyridin-4-yl)-2H-chromen-2-one (6a) 1H and 13C..........................................................................22 6-(Pyridin-4-yl)-2H-chromen-2-one (6b) 1H and 13C .........................................................................23 7-(Pyridin-4-yl)-2H-chromen-2-one (6c) 1H and 13C..........................................................................24 8-(Pyridin-4-yl)-2H-chromen-2-one (6d) 1H and 13C .........................................................................25 2 5-(4-(Trifluoromethyl)phenyl)-2H-chromen-2-one (7a) 1H, 13C and 18F ............................................26 7-(4-(Trifluoromethyl)phenyl)-2H-chromen-2-one (7c) 1H, 13C and 18F ............................................28 8-(4-(Trifluoromethyl)phenyl)-2H-chromen-2-one (7d) 1H, 13C and 18F............................................30 5-(4-Nitrophenyl)-2H-chromen-2-one (8a) 1H and 13C.......................................................................32 7-(4-Nitrophenyl)-2H-chromen-2-one (8c) 1H and 13C.......................................................................33 8-(4-Nitrophenyl)-2H-chromen-2-one (8d) 1H and 13C ......................................................................34 5-(4-Aminophenyl)-2H-chromen-2-one (9a) 1H and 13C....................................................................35 6-(4-Aminophenyl)-2H-chromen-2-one (9b) 1H and 13C....................................................................36 7-(4-Aminophenyl)-2H-chromen-2-one (9c) 1H and 13C ....................................................................37 8-(4-Aminophenyl)-2H-chromen-2-one (9d) 1H and 13C....................................................................38 5-Cyclopropyl-2H-chromen-2-one (10a) 1H and 13C ..........................................................................39 7-Cyclopropyl-2H-chromen-2-one (10c) 1H and 13C..........................................................................40 8-Cyclopropyl-2H-chromen-2-one (10d) 1H and 13C..........................................................................41 6-Amino-3-methyl-2H-chromen-2-one (11) 1H, 13C, IR, HRMS .......................................................42 tButyl (2-oxo-2H-chromen-3-yl)carbamate (Boc-12) 1H and 13C.......................................................44 3-amino-2H-chromen-2-one (12) 1H, 13C, IR, HRMS ........................................................................45