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Supporting Information Discovery and Characterization of (8S, 9R)-5-Fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H- 1,2,4-triazol-5-yl)-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (BMN 673, talazoparib), a Novel, Highly Potent, and Orally Efficacious Poly(ADP-ribose) Polymerase- 1/2 Inhibitor as an Anticancer Agent Bing Wang,* Daniel Chu, Ying Feng, Yuqiao Shen, Mika Aoyagi-Scharber, Leonard E. Post BioMarin Pharmaceutical Inc., 105 Digital Drive, Novato, California 94949 Contents: NMR Spectra of Compound (8S,9R)-47... S2-S5 H-NMR spectrum... S2 C13-NMR spectrum... S3 HMBC-NMR spectrum... S4 HSQC-NMR spectrum... S5 Optical Rotation Measurement for Compound (8S,9R)-47 and (8R, 9S)-47... S6-S7 Single-crystal Structure of Compound (8S,9R)-47... S8-S10 Single-crystal Structure of Compound (2S,3R)-60a... S11-S13 Smiles Strings..... S14-S15 Enantiomeric Purity Analysis of the Compounds in Table 3.... S16-S18 Mouse Body Weight in Xenograft Model Studies..... S19-S20 S1

12.351 7.803 7.720 7.506 7.492 7.485 7.471 7.183 7.160 7.139 7.086 7.080 7.064 7.058 6.937 6.931 6.908 6.903 5.008 5.001 3.660 3.336 2.502-0.001 1.02 0.99 0.94 0.86 1.99 1.99 0.97 2.00 2.99 12 10 8 6 4 2 0 PPM nmrb601, DMSO, F1: 400.132 F2: 1.000 SW1: 8224 OF1: 2474.4 USER: nmrb601 -- DATE: Wed Oct 28 13:19:39 2009 PTS1d: 32768 EX: zg30 PW: 13.8 usec PD: 1.0 sec NA: 8 LB: 0.0 Nuts - $pdata S2

166.119 163.654 163.056 160.624 158.854 158.815 151.936 150.264 148.428 148.295 141.020 135.328 135.300 130.097 130.020 129.904 129.795 115.245 115.035 111.235 102.893 102.626 98.600 98.356 58.576 40.094 42.561 39.883 39.676 39.470 39.254 39.050 38.842 34.830 160 140 120 100 80 60 40 PPM nmrb601, DMSO, F1: 100.623 F2: 1.000 SW1: 24038 OF1: 10012.7 USER: nmrb601 -- DATE: Sun Dec 06 18:25:17 2009 PTS1d: 32768 EX: zgpg30 PW: 8.1 usec PD: 2.0 sec NA: 5000 LB: 0.0 Nuts - $pdata S3

ppm 0 20 40 60 80 100 120 140 160 180 NAME CP-0005007-023-HMBC-DMSO-20091205-T31 EXPNO 10 PROCNO 1 Date_ 20091206 Time 10.43 INSTRUM nmrb601 PROBHD 5 mm PABBO BB- PULPROG hmbcgpndqf TD 4096 SOLVENT DMSO NS 16 DS 16 SWH 4761.905 Hz FIDRES 1.162574 Hz AQ 0.4301300 sec RG 203 DW 105.000 usec DE 6.50 usec TE 295.5 K CNST13 8.0000000 D0 0.00000300 sec D1 1.45412397 sec D6 0.06250000 sec D16 0.00010000 sec IN0 0.00002240 sec ======== CHANNEL f1 ======== NUC1 1H P1 13.78 usec P2 27.56 usec PL1 0.00 db PL1W 11.10253811 W SFO1 400.1329228 MHz ======== CHANNEL f2 ======== NUC2 13C P3 8.06 usec PL2-2.00 db PL2W 54.14257431 W SFO2 100.6228138 MHz ====== GRADIENT CHANNEL ===== GPNAM1 SINE.100 GPNAM2 SINE.100 GPNAM3 SINE.100 GPZ1 50.00 % GPZ2 30.00 % GPZ3 40.10 % P16 1000.00 usec ND0 2 TD 128 SFO1 100.6228 MHz FIDRES 174.386154 Hz SW 221.833 ppm FnMODE QF SI 1024 SF 400.1299968 MHz WDW SINE SSB 0 LB 0.00 Hz GB 0 PC 1.40 SI 1024 MC2 QF SF 100.6128163 MHz WDW SINE SSB 0 LB 0.00 Hz GB 0 200 13 12 11 10 9 8 7 6 5 4 3 2 ppm S4

ppm 13 12 11 10 9 8 7 6 5 4 3 2 ppm S5 0 20 40 60 80 100 120 140 160 180 NAME CP-0005007-023-HSQC-DMSO-20091205-T31 EXPNO 10 PROCNO 1 Date_ 20091206 Time 11.51 INSTRUM nmrb601 PROBHD 5 mm PABBO BB- PULPROG hsqcedetgp TD 1024 SOLVENT DMSO NS 16 DS 16 SWH 4761.905 Hz FIDRES 4.650298 Hz AQ 0.1075700 sec RG 203 DW 105.000 usec DE 6.50 usec TE 295.4 K CNST2 145.0000000 D0 0.00000300 sec D1 1.48832595 sec D4 0.00172414 sec D11 0.03000000 sec D13 0.00000400 sec D16 0.00010000 sec D21 0.00345000 sec IN0 0.00003000 sec ZGOPTNS ======== CHANNEL f1 ======== NUC1 1H P1 13.78 usec P2 27.56 usec P28 1.00 usec PL1 0.00 db PL1W 11.10253811 W SFO1 400.1329228 MHz ======== CHANNEL f2 ======== CPDPRG2 garp NUC2 13C P3 8.06 usec P4 16.12 usec PCPD2 70.00 usec PL2-2.00 db PL12 16.78 db PL2W 54.14257431 W PL12W 0.71703249 W SFO2 100.6202727 MHz ====== GRADIENT CHANNEL ===== GPNAM1 SINE.100 GPNAM2 SINE.100 GPZ1 80.00 % GPZ2 20.10 % P16 1000.00 usec ND0 2 TD 256 SFO1 100.6203 MHz FIDRES 65.104164 Hz SW 165.639 ppm FnMODE Echo-Antiecho SI 1024 SF 400.1299969 MHz WDW QSINE SSB 2 LB 0.00 Hz GB 0 PC 1.40 SI 1024 MC2 echo-antiecho SF 100.6128127 MHz WDW QSINE SSB 2 LB 0.00 Hz GB 0

Optical Rotation of (8S,9R)-47 Report No.: ROA_09000495L04 Project Code: 09000495L04 Sample ID: 09001328 Remark Rotation 0.615 Wavelength Cell Path 589 nm 100 mm Specific Rotation 92.204 Temperature 20 MeOH:CAN:DMF=0.5:0.5: Solvent 1 Instrument RUDOLPH (AUTOPOL V) Automatic Polarimeter Concentration 0.667g/100ml (6.67mg/ml) S6

Optical Rotation of (8R,9S)-47 Report No.: ROA_09000475L04 Project Code: 09000475L04 Sample ID: 09001297 Remark Rotation -0.624 Wavelength Cell Path 589 nm 100 mm Specific Rotation -93.403 Temperature 20 MeOH:ACN:DMF=0.5:0.5: Solvent 1 Instrument RUDOLPH (AUTOPOL V) Automatic Polarimeter Concentration 0.667g/100ml (6.67mg/ml) S7

ORTEP drawing of (8S,9R)-47 Tosylate (LIMS no. 237013). Atoms are represented by 50% probability anisotropic thermal ellipsoids. S8

I. Crystal Data and Data Collection Parameters for (8S,9R)-47 Tosylate formula C 26H 22F 2N 6O 4S formula weight 552.56 space group P2 1 (No. 4) a, Å 11.9122(11) b, Å 8.7136(9) c, Å 12.2781(10) β, deg 99.901(6) V, Å 3 1255.5(2) Z 2 d calc, g cm -3 1.462 crystal dimensions, mm 0.20 x 0.18 x 0.01 temperature, K 150. radiation (wavelength, Å) Cu K α (1.54184) monochromator graphite linear abs coef, mm -1 1.641 absorption correction applied empirical a transmission factors: min, max 0.643, 0.984 diffractometer Rigaku Rapid II h, k, l range -14 to 14-7 to 10-14 to 14 2θ range, deg 13.96-133.11 mosaicity, deg 0.84 programs used SHELXTL F 000 572.0 weighting 1/[σ 2 (F o2 )+(0.0293P) 2 +0.0000P] where P=( F o2 +2F c2 )/3 data collected 14020 unique data 4037 R int 0.077 data used in refinement 4037 cutoff used in R-factor calculations F o2 >2.0σ(F o2 ) data with I>2.0σ(I) 2384 refined extinction coef 0.0026 number of variables 428 largest shift/esd in final cycle 0.00 R(F o) 0.048 R w(f o2 ) 0.077 goodness of fit 0.985 absolute structure determination Flack parameter b (0.04(2)) Hooft parameter c (0.056(19)) Friedel Coverage 82% a CrystalClear: An Integrated Program for the Collection and Processing of Area Detector Data, Rigaku Corporation, 1997-2002. b Flack,H. D. Acta Cryst., 1983 A39, 876. c Hooft,R.W.W., Straver,L.H., and Spek, A.L. J. Appl. Cryst., 2008, 41, 96-103. S9

Positional Parameters and Their Estimated Standard Deviations for (8S,9R)-47 Tosylate Atom x y z U(Å 2 ) S3 0.04429(10) 0.78412(15) 0.25417(9) 0.0355(4) F6-0.38575(19) 0.3765(3) 0.39659(18) 0.0425(8) F24 0.5466(2) 0.4053(3) 0.6503(2) 0.0603(11) O9-0.2948(2) 0.3427(4) 0.0037(2) 0.0450(12) O31 0.0949(2) 0.7002(4) 0.1713(2) 0.0414(10) O32 0.0965(3) 0.9371(4) 0.2779(3) 0.0425(10) O33 0.0447(2) 0.6955(4) 0.3539(2) 0.0400(9) N3 0.0192(3) 0.3364(5) 0.4449(3) 0.0335(12) N10-0.1039(3) 0.3164(5) 0.0419(3) 0.0353(14) N11 0.0033(3) 0.3155(5) 0.1069(3) 0.0349(12) N112 0.3041(3) 0.3601(5) 0.2152(3) 0.0326(12) N113 0.3744(3) 0.2646(6) 0.1683(3) 0.0424(12) N115 0.2199(4) 0.1409(5) 0.1991(3) 0.0313(14) C1 0.1179(4) 0.3484(7) 0.2869(3) 0.0285(16) C2 0.1153(4) 0.2714(7) 0.4007(3) 0.0307(16) C4-0.0864(4) 0.3436(6) 0.3781(3) 0.0317(14) C5-0.1866(4) 0.3581(6) 0.4210(4) 0.0352(16) C6-0.2888(4) 0.3652(6) 0.3496(4) 0.0388(17) C7-0.3022(4) 0.3590(6) 0.2367(4) 0.0358(16) C8-0.2021(4) 0.3465(6) 0.1936(3) 0.0298(14) C9-0.2065(4) 0.3372(6) 0.0734(4) 0.0386(16) C12 0.0041(3) 0.3308(6) 0.2118(3) 0.0316(16) C13-0.0954(3) 0.3408(5) 0.2617(3) 0.0266(10)* C21 0.2279(4) 0.3067(6) 0.4765(3) 0.0274(14) C22 0.2492(5) 0.4453(7) 0.5298(4) 0.0418(19) C23 0.3564(4) 0.4802(7) 0.5910(4) 0.0431(19) C24 0.4403(4) 0.3728(7) 0.5915(4) 0.0391(17) C25 0.4245(5) 0.2349(7) 0.5397(4) 0.045(2) C26 0.3162(4) 0.2001(6) 0.4813(3) 0.0337(16) C31-0.0985(4) 0.8188(6) 0.1965(3) 0.0356(16) C32-0.1230(5) 0.9305(6) 0.1144(4) 0.0439(19) C33-0.2373(5) 0.9607(7) 0.0713(5) 0.052(2) C34-0.3251(5) 0.8836(8) 0.1088(5) 0.059(2) C35-0.2982(5) 0.7737(9) 0.1876(5) 0.060(2) C36-0.1845(5) 0.7408(7) 0.2333(4) 0.047(2) C37-0.4480(7) 0.9274(19) 0.0657(9) 0.095(4) C111 0.2121(4) 0.2862(7) 0.2347(3) 0.0318(16) C112 0.3402(5) 0.5183(6) 0.2425(5) 0.052(2) C114 0.3203(4) 0.1350(7) 0.1590(4) 0.0376(17) H1 0.128(3) 0.456(5) 0.294(3) 0.032(14)* H2 0.114(3) 0.146(5) 0.388(3) 0.029(12)* H3 0.017(3) 0.308(5) 0.511(3) 0.039(13)* H5-0.183(3) 0.352(4) 0.498(3) 0.023(11)* H7-0.376(3) 0.348(6) 0.193(3) 0.056(15)* Starred atoms were refined isotropically U eq = (1/3)Σ iσ j U ija * ia * ja i.a j Hydrogen atoms are included in calculation of structure factors but not refined S10

ORTEP drawing of (2S,3R)-60a. Atoms are represented by 50% probability anisotropic thermal ellipsoids. S11

Crystal Data and Data Collection Parameters for Compound (2S,3R)-60a formula C 20H 16F 2N 4O 3 formula weight 398.37 space group P4 32 12 (No. 96) a, Å 8.8428(2) c, Å 47.153(3) V, Å 3 3687.1(3) Z 8 d calc, g cm -3 1.435 crystal dimensions, mm 0.20 x 0.20 x 0.04 temperature, K 150. radiation (wavelength, Å) Cu K α (1.54184) monochromator graphite linear abs coef, mm -1 0.921 absorption correction applied empirical a transmission factors: min, max 0.769, 0.964 diffractometer Rigaku Rapid II h, k, l range -9 to 8-6 to 10-54 to 55 2θ range, deg 13.14-133.09 mosaicity, deg 2.87 programs used SHELXTL F 000 1648.0 weighting 1/[σ 2 (F o2 )+(0.0461P) 2 +0.5849P] where P=( F o2 +2F c2 )/3 data collected 18568 unique data 3252 R int 0.069 data used in refinement 3252 cutoff used in R-factor calculations F o2 >2.0σ(F o2 ) data with I>2.0σ(I) 2691 refined extinction coef 0.0014 number of variables 327 largest shift/esd in final cycle 0.00 R(F o) 0.053 R w(f o2 ) 0.100 goodness of fit 1.100 absolute structure determination Flack parameter b (-0.1(2)) Hooft parameter c (-0.08(11)) Friedel Coverage 100% a CrystalClear: An Integrated Program for the Collection and Processing of Area Detector Data, Rigaku Corporation, 1997-2002. b Flack,H. D. Acta Cryst., 1983 A39, 876. c Hooft,R.W.W., Straver,L.H., and Spek, A.L. J. Appl. Cryst., 2008, 41, 96-103. S12

Positional Parameters and Estimated Standard Deviations for Compound (2S,3R)-60a Atom x y z U(Å 2 ) F1 0.53477(19) 0.71906(19) 0.24920(3) 0.0480(5) F2 0.47299(19) 0.76574(19) 0.01669(3) 0.0491(5) O1 0.8578(2) 0.3024(2) 0.15876(3) 0.0386(6) O2 0.7834(2) 0.1873(2) 0.21736(4) 0.0407(6) O3 0.9882(2) 0.3376(2) 0.21642(4) 0.0402(6) N1 0.5845(3) 0.6734(3) 0.14943(4) 0.0303(7) N2 0.9516(3) 0.4996(3) 0.09660(4) 0.0333(6) N3 0.9540(3) 0.2712(3) 0.07469(5) 0.0438(8) N4 0.8322(3) 0.2841(3) 0.09263(4) 0.0341(7) C1 0.5972(3) 0.6469(3) 0.22648(5) 0.0344(8) C2 0.5600(3) 0.6980(4) 0.20006(5) 0.0331(8) C3 0.6231(3) 0.6238(3) 0.17651(5) 0.0286(8) C4 0.6871(3) 0.6342(3) 0.12608(5) 0.0275(7) C5 0.7217(3) 0.4648(3) 0.12742(5) 0.0266(7) C6 0.7757(3) 0.4141(3) 0.15663(5) 0.0287(7) C7 0.7188(3) 0.4987(3) 0.18081(5) 0.0276(7) C8 0.7519(3) 0.4506(3) 0.20874(5) 0.0301(8) C9 0.8412(3) 0.3108(3) 0.21417(5) 0.0329(8) C10 1.0796(5) 0.2060(4) 0.22220(9) 0.0543(10) C11 0.6926(3) 0.5264(3) 0.23179(6) 0.0348(8) C12 0.8330(3) 0.4187(3) 0.10542(5) 0.0283(7) C13 1.0206(4) 0.4033(4) 0.07803(6) 0.0395(9) C14 0.7307(5) 0.1565(4) 0.09625(8) 0.0508(12) C15 0.6204(3) 0.6781(3) 0.09770(5) 0.0264(7) C16 0.6962(3) 0.7787(3) 0.08039(5) 0.0306(8) C17 0.6470(4) 0.8104(3) 0.05301(6) 0.0364(8) C18 0.5190(3) 0.7404(3) 0.04381(5) 0.0348(8) C19 0.4376(3) 0.6414(3) 0.06017(6) 0.0361(9) C20 0.4887(3) 0.6108(3) 0.08751(5) 0.0338(8) H1 0.546(4) 0.783(5) 0.1491(7) 0.091(13)* H2 0.493(3) 0.785(3) 0.1973(5) 0.038(8)* H4 0.782(3) 0.687(3) 0.1289(5) 0.023(7)* H5 0.632(3) 0.414(3) 0.1240(5) 0.035(8)* H11 0.721(3) 0.500(3) 0.2522(6) 0.045(8)* H13 1.125(4) 0.424(3) 0.0676(6) 0.068(10)* H16 0.793(3) 0.826(3) 0.0870(6) 0.052(9)* H17 0.697(4) 0.879(4) 0.0413(6) 0.061(10)* H19 0.346(4) 0.593(4) 0.0532(6) 0.061(10)* H20 0.435(4) 0.536(4) 0.0995(5) 0.054(9)* H10A 1.058(4) 0.159(4) 0.2421(7) 0.071(11)* H10B 1.185(4) 0.243(4) 0.2231(6) 0.061(10)* H10C 1.068(4) 0.142(4) 0.2058(7) 0.071(12)* H14A 0.770(5) 0.080(5) 0.1101(9) 0.116(16)* H14B 0.739(5) 0.103(5) 0.0779(8) 0.106(15)* H14C 0.625(6) 0.184(6) 0.0968(9) 0.131(19)* Starred atoms were refined isotropically U eq = (1/3)Σ iσ j U ija * ia * ja i.a j Hydrogen atoms are included in calculation of structure factors but not refined S13

Compound Smiles PARP-1 Enzyme Cellular PAR TMZ Chemosensitizationᵈ Capan-1 rlm IC₅₀ (nm)ᵃ inhibition GI₅₀ (µm)ᵇ (BRCA2 mutant) T1/2 EC₅₀ (nm)ᵇ EC₅₀ (µm)ᵇ (min)ᵃ 13 O=C1NN=C2C(C3=CC=CC=C3)C(C4=CC=CC=C4)NC5=CC=CC1=C52 6.1 58.4 324 2.97 24 O=C1NN=C2C(C3=CC=C(CN(C)C)C=C3)C(C4=CC=CC=C4)NC5=CC=CC1=C52 5.85 112 6 25 O=C1NN=C2C(C3=CC=CC=C3)C(C4=CC=C(CN(C)C)C=C4)NC5=CC=CC1=C52 2.36 6 59 0.387 12 26 O=C1NN=C2C(C3=CC=CC(CN(C)C)=C3)C(C4=CC=CC=C4)NC5=CC=CC1=C52 3.65 204 11 27 O=C1NN=C2C(C3=CC=CC=C3)C(C4=CC(CN(C)C)=CC=C4)NC5=CC=CC1=C52 2.63 128 19 O=C1NN=C2C(C3=CC=C(CN(C)C)C=C3)C(C4=CC=C(CN(C)C)C=C4)NC5=CC=CC1=C52 3.89 18.6 63 0.979 >120 28 O=C1NN=C2C(C3=CC=C(C(C)C)C=C3)C(C4=CC=C(CN(C)C)C=C4)NC5=CC=CC1=C52 9.59 15.2 123 29 O=C1NN=C2C(C3=CC=C(C)C=C3)C(C4=CC=C(CN(C)C)C=C4)NC5=CC=CC1=C52 1.95 10.8 123 0.514 35 30 O=C1NN=C2C(C3=CC=C(C(F) (F)F)C=C3)C(C4=CC=C(CN(C)C)C=C4)NC5=CC=CC1=C52 6.29 6.7 94 0.818 52 O=C1NN=C2C(C3=CC=C(F)C=C3)C(C4=CC=C(CN(C)C)C=C4)NC5=CC=CC1=C52 2.48 18.5 76 1.73 44 31 O=C1NN=C2C(C3=CC=CC=C3)C(C4=CC=C(CN(C)C)C=C4)NC5=CC(F)=CC1=C52 3.29 8.48 46 0.146 5 32 O=C1NN=C2C(C3=CC=C(F)C=C3)C(C4=CC=C(CN(C)C)C=C4)NC5=CC(F)=CC1=C52 2.63 6.1 94 0.134 82 53 O=C1NN=C2C(C3=CC=CC=C3)C(C4=CC=C(CN5CCN(C)CC5)C=C4)NC6=CC=CC1=C62 2.02 40.6 54.6 0.603 19 54 O=C1NN=C2C(C3=CC=CC=C3)C(C4=CC=C(CN5CCCC5)C=C4)NC6=CC=CC1=C62 4.02 9.47 90 0.298 55 O=C1NN=C2C(C3=CC=CC=C3)C(C4=CC=C(CN5CCC5)C=C4)NC6=CC=CC1=C62 3.63 9.7 89 0.434 8 56 O=C1NN=C2C(C3=CC=C(F)C=C3)C(C4=CC=C(CN5CCCC5)C=C4)NC6=CC=CC1=C62 2.02 23 134 0.454 57 O=C1NN=C2C(C3=CC=C(F)C=C3)C(C4=CC=C(CN5CCC5)C=C4)NC6=CC=CC1=C62 3.09 3.81 95 0.276 89 15 O=C1NN=C2C(C3=CC=NC=C3)C(C4=CC=NC=C4)NC5=CC=CC1=C52 22.4 >400 16 O=C1NN=C2C(C3=CC=CC=N3)C(C4=CC=CC=N4)NC5=CC=CC1=C52 108 17 O=C1NN=C2C(C3=CC=CN=C3)C(C4=CN=CC=C4)NC5=CC=CC1=C52 17.7 18 O=C1NN=C2C(C3=NC=CN3C)C(C4=NC=CN4C)NC5=CC=CC1=C52 24.5 20 O=C1NN=C2C(C3CCCNC3)C(C4=CN=CC=C4)NC5=CC=CC1=C52 105 33 O=C1NN=C2C(C(C)C)C(C3=CC=CC=C3)NC4=CC=CC1=C42 4139 34 O=C1NN=C2C(C3=CC=CC=C3)C(C4=NC=CN4C)NC5=CC=CC1=C52 9.45 >400 35 O=C1NN=C2C(C3=NC=CN3C)C(C4=CC=CC=C4)NC5=CC=CC1=C52 2.35 16.9 97 0.36 89 36 O=C1NN=C2C(C3=NC=CN3C)C(C4=CC=C(F)C=C4)NC5=CC=CC1=C52 2.08 19.7 75.5 0.418 >120 37 O=C1NN=C2C(C3=NC=CN3C)C(C4=CC=CC=C4)NC5=CC(F)=CC1=C52 2.14 18.1 36.3 0.108 103 38 O=C1NN=C2C(C3=NC=CS3)C(C4=CC=C(F)C=C4)NC5=CC=CC1=C52 7.35 24.4 106 39 O=C1NN=C2C(C3=CC=NN3C)C(C4=CC=C(F)C=C4)NC5=CC=CC1=C52 1.84 9.33 > 400 0.956 40 O=C1NN=C2C(C3=NC=CN3C)C(C4=CC=C(C(F)(F)F)C=C4)NC5=CC=CC1=C52 5.27 27.2 192 1.1 41 O=C1NN=C2C(C3=NC=CN3C)C(C4=CC=CC=C4)NC5=CC(Cl)=CC1=C52 6.18 121 111 0.785 42 O=C1NN=C2C(C3=NC=CN3C)C(C4=CC=C(F)C=C4)NC5=CC(F)=CC1=C52 1.87 17 35.7 0.056 43 O=C1NN=C2C(C3=NC=NN3C)C(C4=CC=CC=C4)NC5=CC=CC1=C52 2.4 149 > 400 0.17 414 44 O=C1NN=C2C(C3=NC=NN3C)C(C4=CC=CC=C4)NC5=CC(F)=CC1=C52 2.6 24 0.009 472 45 O=C1NN=C2C(C3=NN=CN3C)C(C4=CC=CC=C4)NC5=CC=CC1=C52 2.76 > 400 4.34 46 O=C1NN=C2C(C3=NC=NN3C)C(C4=CC=C(F)C=C4)NC5=CC=CC1=C52 2.29 6.94 44 0.071 >120 47 O=C1NN=C2C(C3=NC=NN3C)C(C4=CC=C(F)C=C4)NC5=CC(F)=CC1=C52 2.14 5.48 19 0.008 359 61 O=C1NN=C2C(C3=NC=NN3C)C(C4=CC=C(NC(C)=O)C=C4)NC5=CC(F)=CC1=C52 2.08 0.146 >120 62 O=C1NN=C2C(C3=NC=NN3C)C(C4=CN=CN=C4)NC5=CC(F)=CC1=C52 7.62 63 O=C1NN=C2C(C3=NC=NN3C)C(C4=CC=NN4C)NC5=CC(F)=CC1=C52 2.88 1.92 64 O=C1NN=C2C(C3=NC=NN3C)C(C4=CC=C(C#N)C=C4)NC5=CC(F)=CC1=C52 3.9 0.088 4 (Rucaparib) FC1=CC2=C3C(NC(C4=CC=C(CNC)C=C4)=C3CCNC2=O)=C1 1.98 4.74 67 0.609 ᵃ Mean of duplicate determinations. ᵇ Mean of triplicate determinations. ᵈ TMZ sensitization condition: the concentration (µm) of TMZ that kills 50% of the treated cells (GI50) in the presence of 0.4 µm of PARP inhibitors S14

Compound Smiles PARP-1 Cellular PAR TMZ Chemo- Capan-1 Enzyme inhibition sensitizationᵈ (BRCA2 deficient) IC₅₀ (nm)ᵃ EC₅₀ (nm)ᵇ GI₅₀ (µm)ᵇ EC₅₀ (nm)ᵇ (8R,9S)-36 O=C1NN=C2[C@@H](C3=NC=CN3C)[C@H](C4=CC=C(F)C=C4)NC5=CC=CC1=C52 136 285 > 10,000 6310 (8S,9R)-36 O=C1NN=C2[C@H](C3=NC=CN3C)[C@@H](C4=CC=C(F)C=C4)NC5=CC=CC1=C52 2.25 6.9 261 305 (8R,9S)-42 O=C1NN=C2[C@@H](C3=NC=CN3C)[C@H](C4=CC=C(F)C=C4)NC5=CC(F)=CC1=C52 9.5 164 104 124 (8S,9R)-42 O=C1NN=C2[C@H](C3=NC=CN3C)[C@@H](C4=CC=C(F)C=C4)NC5=CC(F)=CC1=C52 1.6 17.9 25 39 (8S,9R)-46 O=C1NN=C2[C@H](C3=NN=CN3C)[C@@H](C4=CC=C(F)C=C4)NC5=CC=CC1=C52 1.46 5.1 41 37 (8R,9S)-46 O=C1NN=C2[C@@H](C3=NN=CN3C)[C@H](C4=CC=C(F)C=C4)NC5=CC=CC1=C52 68.9 250 3263 1883 (8S,9R)-57 O=C1NN=C2[C@H](C3=CC=C(F)C=C3)[C@@H](C4=CC=C(CN5CCC5)C=C4)NC6=CC=CC1=C62 1.1 6.3 279 389 (8R,9S)-57 O=C1NN=C2[C@@H](C3=CC=C(F)C=C3)[C@H](C4=CC=C(CN5CCC5)C=C4)NC6=CC=CC1=C62 4.4 11.8 3572 2249 (8S,9R)-47 O=C1NN=C2[C@H](C3=NC=NN3C)[C@@H](C4=CC=C(F)C=C4)NC5=CC(F)=CC1=C52 0.57 2.51 4 5 (8R,9S)-47 O=C1NN=C2[C@@H](C3=NC=NN3C)[C@H](C4=CC=C(F)C=C4)NC5=CC(F)=CC1=C52 144 864 1807 1135 ᵃ Mean of duplicate determinations. ᵇ Mean of triplicate determinations. ᵈ TMZ sensitization condition: the concentration (nm) of a PARP inhibitor that kills 50% of the treated cells (GI50) in the presence of 200 µm of TMZ is reported. S15

Enantiomeric Purity Analysis of the Compounds in Table 3 S16

Enantiomeric Purity Analysis of the Compounds in Table 3 (cont d) S17

Enantiomeric Purity Analysis of the Compounds in Table 3 (cont d) S18

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