Chemogenomic set

Screened at 10 µM (KINOMEscan, DiscoverX), off-target with >90% PoC; IC50(CDK4/7) = 220/200 nM (Radiometric kinase assay,https://pubs.acs.org/doi/10.1021/acs.jmedchem.8b00049);
In-vitro potency (radiometric kinase assay, using ATP = Km): IC50(CDK1/cyclinB) = 26 nM, IC50(CDK2/cyclinE) = 3 nM, IC50(CDK2/cyclinA) = 4 nM, IC50(CDK4/cyclinD) = 216 nM, IC50(CDK5/p25) = 10 nM, IC50(CDK7/cyclinH) = 200 nM, IC50(CDK9/cyclinT)= 13 nM, https://pubs.acs.org/doi/10.1021/acs.jmedchem.8b00049;

Screened at 1 µM, closest targets as % of control: NEK1(45%), STK38L (47%), BMX(50%)

Screened at 1 µM, closest targets as % of control: NEK1(45%), STK38L (47%), BMX(50%)

Various assays types used, i.e. TR-FRET, Luminescence assay, FLINT, Electrophysiological assay, LEADseeker SPA; No activation across HDACs 1–11 at 100 μM; Selective for PADI4 over PADI1-3 (tested with recombinant enzymes and cell lysates).

Screened at 1 µM, closest target as % of inhibition: CHRM2 (40.7%)

inactive at 10 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

Screened at 3 µM, in-vitro potency of closest target: Kd(CSF1R) = 2716 nM (90.1% inhibition), S-score(10) = 0.005 at 3 µM; In-vitro potencies of other FGFR family members (enzyme activity assays): IC50(FGFR1) = 591 nM, IC50(FGFR2) = 493 nM, IC50(FGFR3) = 150 nM; IC50(p70S6Kb) > 10 μM (https://aacr.silverchair-cdn.com/aacr/content_public/journal/cancerdiscovery/5/4/10.1158_2159-8290.cd-14-1029/4/21598290cd141029-sup-137537_2_supp_0_njmz90.pdf?Expires=1675264845&Signature=C7J4HfbPNEkAbAqzPJ39H9yL-DM-ZriAd8q9~z8YUG1gWwYiv~YY3KnIOiETJu6s6ant4xiTBoYwoCLgll18xYBmbGpqFihOfhV-o13a7i6vEvF1O1eaowwAMUXKijlCNqDlMeXH0BaQ1JGUfaWJ-rcaUZdvfAS~UHg~9yVyE3t7x2JTKWLV3T1OQ0xNVih6dxAH~ACl87xwuIVD4NVAlkEcGUWPGXNzCUzk8-SlPnWgBxnUZLP6R8NhMbrjeOlLoYg0TNe9HVYyYuOCI6bAajPrnqSaYv1qC1eoQq-HWafjwJ7N3T5waFSlCi~COA3D0QYQ~UdoNRFASuZZzqG1Lg__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)

Screened at 20 µM, closest targets in the screen: dTm FLT1 = 7.6 K, STK10 = 4.8K, clean profile;
Screened in enzymatic assays: IC50(PDGFR) >50 µM, IC50(EGFR) >50 µM, IC50(SRC) >50 µM, IC50(MAPK1) >50 µM, IC50(PRKCA) >50 µM, IC50(INSR) >50 µM, IC50(CDK4) >50 µM; In-cellulo follow-up of closest targets (autophosphorylation of PDGFR in rat aortic smooth muscle cells, EGF receptors in A431 cells, and INSR in NIHIR cells): IC50(EGFR) >1 µM, IC50(INSR) >1 µM, IC50(PDGF) >30 µM (https://jpet.aspetjournals.org/content/286/1/569.long);

Screened at 1 μM, closest targets as % of inhibition: FLT3 (96%), GSG2 (93%), TYK2 (93%), CSNK2A (66%), KIT(A829) = 79%), KIT(D816) = 86%), MINK (77%), PDGFRA (79%), PDGFRB (57%), MELK (86%); In-vitro follow-up of closest targets (biochemical assays): IC50(FLT3) = 180 nM, IC50(GSG2) = 190 nM, IC50(KIT, A829) > 10 µM, IC50(PDGFRA) = 420 nM, IC50(MELK) = 2 nM (https://pubs.acs.org/doi/10.1021/acs.jmedchem.6b00052);

Screened at 1 μM inhibition, closest targets as % of residual activity: MAP4K5 (8%), PAK3 (9%), PAK2 (10%), NLK (13%), PKN3 (35%), PAK1 (36%), MAP2K4 (37%), TIE2 (39%), MST4 (45%), MELK (48%); In-vitro follow-up of closest targets (33PanQinase assay, Reaction Biology): IC50(MAP4K5) = 210 nM, IC50(PAK3) = 140 nM, IC50(PAK2) = 41 nM, IC50(NLK) = 13 nM, IC50(PKN3) = 1400 nM, IC50(PAK1) = 580 nM, IC50(MAP2K4) = 830 nM, IC50(TIE2) = 3100 nM, IC50(MST4) = 1600 nM, IC50(MELK) = 2200 nM (https://pubs.acs.org/doi/10.1021/acs.jmedchem.0c02144); In-cellulo follow-up of closest targets (NanoBRET assay in HEK293T cells): IC50(MAP4K5) = 13 µM, IC50(NLK) = 0.25 µM, IC50(PKN3) = 6.7 µM, IC50(TIE2) = 6 µM, IC50(MST4) = 34 µM, https://pubs.acs.org/doi/10.1021/acs.jmedchem.0c02144;

Screened at 1 μM inhibition, closest targets as % of residual activity: MAP4K5 (8%), PAK3 (9%), PAK2 (10%), NLK (13%), PKN3 (35%), PAK1 (36%), MAP2K4 (37%), TIE2 (39%), MST4 (45%), MELK (48%); In-vitro follow-up of closest targets (33PanQinase assay, Reaction Biology): IC50(MAP4K5) = 210 nM, IC50(PAK3) = 140 nM, IC50(PAK2) = 41 nM, IC50(NLK) = 13 nM, IC50(PKN3) = 1400 nM, IC50(PAK1) = 580 nM, IC50(MAP2K4) = 830 nM, IC50(TIE2) = 3100 nM, IC50(MST4) = 1600 nM, IC50(MELK) = 2200 nM (https://pubs.acs.org/doi/10.1021/acs.jmedchem.0c02144); In-cellulo follow-up of closest targets (NanoBRET assay in HEK293T cells): IC50(MAP4K5) = 13 µM, IC50(NLK) = 0.25 µM, IC50(PKN3) = 6.7 µM, IC50(TIE2) = 6 µM, IC50(MST4) = 34 µM, https://pubs.acs.org/doi/10.1021/acs.jmedchem.0c02144;

Screened at 1 μM inhibition, closest targets as % of residual activity: MAP4K5 (8%), PAK3 (9%), PAK2 (10%), NLK (13%), PKN3 (35%), PAK1 (36%), MAP2K4 (37%), TIE2 (39%), MST4 (45%), MELK (48%); In-vitro follow-up of closest targets (33PanQinase assay, Reaction Biology): IC50(MAP4K5) = 210 nM, IC50(PAK3) = 140 nM, IC50(PAK2) = 41 nM, IC50(NLK) = 13 nM, IC50(PKN3) = 1400 nM, IC50(PAK1) = 580 nM, IC50(MAP2K4) = 830 nM, IC50(TIE2) = 3100 nM, IC50(MST4) = 1600 nM, IC50(MELK) = 2200 nM (https://pubs.acs.org/doi/10.1021/acs.jmedchem.0c02144); In-cellulo follow-up of closest targets (NanoBRET assay in HEK293T cells): IC50(MAP4K5) = 13 µM, IC50(NLK) = 0.25 µM, IC50(PKN3) = 6.7 µM, IC50(TIE2) = 6 µM, IC50(MST4) = 34 µM, https://pubs.acs.org/doi/10.1021/acs.jmedchem.0c02144;

Clean selectivity profile with no target activity >50%; Screened in enzymatic assays against other transaminases: IC50(GOT1/2) >50 µM; Screened at 6 µM against 30 proteases, all targets IC50 >10 µM; Screened at 2 µM against 45 kinases (enzymatic assays), clean selectivity profile with all kinases IC50 >7 µM

Clean selectivity profile with no target activity >50%; Screened in enzymatic assays against other transaminases: IC50(GOT1/2) >50 µM; Screened at 6 µM against 30 proteases, all targets IC50 >10 µM; Screened at 2 µM against 45 kinases (enzymatic assays), clean selectivity profile with all kinases IC50 >7 µM

Screened at 10 µM, in vitro-potencies of closest targets: Ki(SLC6A3) = 1377.15 nM, Ki(HTR2A) = 2576.32 nM; Selectivity against family members in biochemical assays, all >30 fold: IC50(hTRPV1) >25 µM, IC50(hTRPV4) >25 µM, IC50(hTRPC3) >25 µM, IC50(hTRPC5) = 5.6 µM, IC50(hTRPC6) >25 µM, IC50(hKCNK9, TASK-3) >30 µM, IC50(hCACNA1H, Cav3.2) >25 µM; Selectivity screen in Eurofins Lead Profiling Screen, GPCR Profiling Screen and Bayer Kinase Panel was performed, in-vitro potencies of closest targets: Ki (SLC6A3, human) = 0.9 μM, Ki(PGR, human) = 4 μM, EC50(ESR1) = 2.1 μM

Screened at 10 µM, closest target as % of inhibition: HTR3A (38%); Screened against other human prostanoid receptor (binding assay, Eurofins/Panlabs): IC50(PTGFR) = 22 nM, IC50(EP1) >10 µM, IC50(EP2) >10 µM, IC50(EP3) >10 µM, IC50(EP4) >10 µM, IC50(IP) >10 µM, IC50(DP) >10 µM, IC50(CRTH2) >10 µM, >420-fold selective; Screened in cell-based assay (Eurofins/Panlabs): IC50(TBXA2R) = 2.2 µM, 200-fold selective; Screened at 10 µM against 77 other targets (Leadprofiling Screen, Panlabs), closest target as % of inhibition: TBXAS1 (33%); Screened at 10 µM against 8 ion channels (Ion channel profiler, Eurofins): IC20(Nav1.5) >10 μM, IC20(Kv4.3/KChIP2) >10 μM, IC20(Cav1.2) >10 μM, IC20(Kv1.5) >10 μM, IC20(KCNQ/minK) >10 μM, IC20(herG) >10 μM, IC20(HCN4) >10 μM, IC20(Kir2.1) >10 μM

Screened at 10 µM, in-vitro potency of closest target: IC50(CTSG, cathepsin G) = 110 nM, >100-fold selective; Screened at 10 µM against 44 targets outside target family (Eurofins Safety Screen44TM), clean selectivity profile; Screened at 10 µM against 45 GPCRs (PDSP screen), in-vitro potencies of closest targets: Ki(HTR1D) = 1130.61 nM, Ki(OPRK1) = 4948.48 nM

In-vitro potencies of closest targets in BROMOScan (DiscoverX): pKd(TAF1, BD2) = 6.4, pKd(TAF1L, BD2) = 5.7, pKd(BRD9) = 5.7, pKd(CECR2) = 5.5, pKd(BAZ2A) = 5.5, pKd(BAZ2B) = 5.1; Selective for the BD2 domain of BET proteins; Clean in selectivity screen against 48 targets (GPCRs, ion-channels, other targets); Chemoproteomics data (3 vectors) showed excellent non BCPs selectivity; Screened at 10 µM against 45 GPCRs (PDSP screen), in-vitro potencies of closest targets: Ki(DRD3) = 257.47 nM, Ki(DRD4) = 982.65 nM, Ki(DRD2) = 964.05 nM

In-vitro potencies of closest targets in BROMOScan (DiscoverX): pKd(TAF1, BD2) = 6.4, pKd(TAF1L, BD2) = 5.7, pKd(BRD9) = 5.7, pKd(CECR2) = 5.5, pKd(BAZ2A) = 5.5, pKd(BAZ2B) = 5.1; Selective for the BD2 domain of BET proteins; Clean in selectivity screen against 48 targets (GPCRs, ion-channels, other targets); Chemoproteomics data (3 vectors) showed excellent non BCPs selectivity; Screened at 10 µM against 45 GPCRs (PDSP screen), in-vitro potencies of closest targets: Ki(DRD3) = 257.47 nM, Ki(DRD4) = 982.65 nM, Ki(DRD2) = 964.05 nM

In-vitro potencies of closest targets in BROMOScan (DiscoverX): pKd(TAF1, BD2) = 6.4, pKd(TAF1L, BD2) = 5.7, pKd(BRD9) = 5.7, pKd(CECR2) = 5.5, pKd(BAZ2A) = 5.5, pKd(BAZ2B) = 5.1; Selective for the BD2 domain of BET proteins; Clean in selectivity screen against 48 targets (GPCRs, ion-channels, other targets); Chemoproteomics data (3 vectors) showed excellent non BCPs selectivity; Screened at 10 µM against 45 GPCRs (PDSP screen), in-vitro potencies of closest targets: Ki(DRD3) = 257.47 nM, Ki(DRD4) = 982.65 nM, Ki(DRD2) = 964.05 nM

In-vitro potencies of closest targets in BROMOScan (DiscoverX): pKd(TAF1, BD2) = 6.4, pKd(TAF1L, BD2) = 5.7, pKd(BRD9) = 5.7, pKd(CECR2) = 5.5, pKd(BAZ2A) = 5.5, pKd(BAZ2B) = 5.1; Selective for the BD2 domain of BET proteins; Clean in selectivity screen against 48 targets (GPCRs, ion-channels, other targets); Chemoproteomics data (3 vectors) showed excellent non BCPs selectivity; Screened at 10 µM against 45 GPCRs (PDSP screen), in-vitro potencies of closest targets: Ki(DRD3) = 257.47 nM, Ki(DRD4) = 982.65 nM, Ki(DRD2) = 964.05 nM

In-vitro potencies of closest targets in BROMOScan (DiscoverX): Kd(BRD2, BD1) = 1621 nM, Kd(BRD2, BD2) = 35 nM (64-fold selective), Kd(BRD3, BD1) = 2082 nM, Kd(BRD3, BD2) = 32 nM (63-fold selective), Kd(BRD4, BD1) = 769 nM, Kd(BRD4, BD2) = 9 nM (85-fold selective), Kd(BRDT, BD1) = 2454 nM, Kd(BRDT, BD2) = 15 nM (164-fold selective); In-vitro potencies (SPR measurements): Kd(BRD4, BD1) >10 µM, Kd(BRD4, BD2) = 30 nM, excellent selectivity against other BCPs, Kd >30 µM for TAF1(BD2), BRD9, TAF1L(BD2), BAZ2A, CECR2, BAZ2B; Screened at 10 µM against 45 GPCRs (PDSP screen), in-vitro potency of closest target: Ki(HTR1D) = 3438.75 nM

In-vitro potencies of closest targets in BROMOScan (DiscoverX): Kd(BRD2, BD1) = 1621 nM, Kd(BRD2, BD2) = 35 nM (64-fold selective), Kd(BRD3, BD1) = 2082 nM, Kd(BRD3, BD2) = 32 nM (63-fold selective), Kd(BRD4, BD1) = 769 nM, Kd(BRD4, BD2) = 9 nM (85-fold selective), Kd(BRDT, BD1) = 2454 nM, Kd(BRDT, BD2) = 15 nM (164-fold selective); In-vitro potencies (SPR measurements): Kd(BRD4, BD1) >10 µM, Kd(BRD4, BD2) = 30 nM, excellent selectivity against other BCPs, Kd >30 µM for TAF1(BD2), BRD9, TAF1L(BD2), BAZ2A, CECR2, BAZ2B; Screened at 10 µM against 45 GPCRs (PDSP screen), in-vitro potency of closest target: Ki(HTR1D) = 3438.75 nM

In-vitro potencies of closest targets in BROMOScan (DiscoverX): Kd(BRD2, BD1) = 1621 nM, Kd(BRD2, BD2) = 35 nM (64-fold selective), Kd(BRD3, BD1) = 2082 nM, Kd(BRD3, BD2) = 32 nM (63-fold selective), Kd(BRD4, BD1) = 769 nM, Kd(BRD4, BD2) = 9 nM (85-fold selective), Kd(BRDT, BD1) = 2454 nM, Kd(BRDT, BD2) = 15 nM (164-fold selective); In-vitro potencies (SPR measurements): Kd(BRD4, BD1) >10 µM, Kd(BRD4, BD2) = 30 nM, excellent selectivity against other BCPs, Kd >30 µM for TAF1(BD2), BRD9, TAF1L(BD2), BAZ2A, CECR2, BAZ2B; Screened at 10 µM against 45 GPCRs (PDSP screen), in-vitro potency of closest target: Ki(HTR1D) = 3438.75 nM

In-vitro potencies of closest targets in BROMOScan (DiscoverX): Kd(BRD2, BD1) = 1621 nM, Kd(BRD2, BD2) = 35 nM (64-fold selective), Kd(BRD3, BD1) = 2082 nM, Kd(BRD3, BD2) = 32 nM (63-fold selective), Kd(BRD4, BD1) = 769 nM, Kd(BRD4, BD2) = 9 nM (85-fold selective), Kd(BRDT, BD1) = 2454 nM, Kd(BRDT, BD2) = 15 nM (164-fold selective); In-vitro potencies (SPR measurements): Kd(BRD4, BD1) >10 µM, Kd(BRD4, BD2) = 30 nM, excellent selectivity against other BCPs, Kd >30 µM for TAF1(BD2), BRD9, TAF1L(BD2), BAZ2A, CECR2, BAZ2B; Screened at 10 µM against 45 GPCRs (PDSP screen), in-vitro potency of closest target: Ki(HTR1D) = 3438.75 nM

In-vitro potencies of closest targets in BROMOScan (DiscoverX): Kd(BRD2, BD1) = 13 nM, Kd(BRD3, BD1) = 5 nM, Kd(BRD4, BD1) = 5.9 nM, Kd(BRDT, BD1) = 18 nM; Selectivity screen against 50 targets (GPCRs, ion-channels, other targets), in-vitro potencies of closest targets: pIC50(CHRNA1) = 6, pIC50(CYP3A4) = 6; Screened at 10 µM against 45 GPCRs (PDSP screen), in-vitro potencies of closest targets: Ki(DRD3) = 485.92 nM, Ki(DRD4) = 856.99 nM, Ki(GABAA) = 1595.89 nM, Ki(GABAA/BZP) = 1970.29 nM

In-vitro potencies of closest targets in BROMOScan (DiscoverX): Kd(BRD2, BD1) = 13 nM, Kd(BRD3, BD1) = 5 nM, Kd(BRD4, BD1) = 5.9 nM, Kd(BRDT, BD1) = 18 nM; Selectivity screen against 50 targets (GPCRs, ion-channels, other targets), in-vitro potencies of closest targets: pIC50(CHRNA1) = 6, pIC50(CYP3A4) = 6; Screened at 10 µM against 45 GPCRs (PDSP screen), in-vitro potencies of closest targets: Ki(DRD3) = 485.92 nM, Ki(DRD4) = 856.99 nM, Ki(GABAA) = 1595.89 nM, Ki(GABAA/BZP) = 1970.29 nM

In-vitro potencies of closest targets in BROMOScan (DiscoverX): Kd(BRD2, BD1) = 13 nM, Kd(BRD3, BD1) = 5 nM, Kd(BRD4, BD1) = 5.9 nM, Kd(BRDT, BD1) = 18 nM; Selectivity screen against 50 targets (GPCRs, ion-channels, other targets), in-vitro potencies of closest targets: pIC50(CHRNA1) = 6, pIC50(CYP3A4) = 6; Screened at 10 µM against 45 GPCRs (PDSP screen), in-vitro potencies of closest targets: Ki(DRD3) = 485.92 nM, Ki(DRD4) = 856.99 nM, Ki(GABAA) = 1595.89 nM, Ki(GABAA/BZP) = 1970.29 nM

In-vitro potencies of closest targets in BROMOScan (DiscoverX): Kd(BRD2, BD1) = 13 nM, Kd(BRD3, BD1) = 5 nM, Kd(BRD4, BD1) = 5.9 nM, Kd(BRDT, BD1) = 18 nM; Selectivity screen against 50 targets (GPCRs, ion-channels, other targets), in-vitro potencies of closest targets: pIC50(CHRNA1) = 6, pIC50(CYP3A4) = 6; Screened at 10 µM against 45 GPCRs (PDSP screen), in-vitro potencies of closest targets: Ki(DRD3) = 485.92 nM, Ki(DRD4) = 856.99 nM, Ki(GABAA) = 1595.89 nM, Ki(GABAA/BZP) = 1970.29 nM

In-vitro potencies of closest targets in BROMOScan (DiscoverX): pKd(TAF1, BD2) = 7.3, pKd(TAF1L, BD2) = 6.4, selective for the BD1 domain of BET proteins; Follow-up of closest targets (FRET assay): pIC50(TAF1, BD1) = 5.0 ± 0.1, n = 3 (BRD4, BD1 selectivity = 500-fold); Screened at 10 µM against 45 GPCRs (PDSP screen), in-vitro potency of closest target: Ki(DRD4) = 1013.12 nM

In-vitro potencies of closest targets in BROMOScan (DiscoverX): pKd(TAF1, BD2) = 7.3, pKd(TAF1L, BD2) = 6.4, selective for the BD1 domain of BET proteins; Follow-up of closest targets (FRET assay): pIC50(TAF1, BD1) = 5.0 ± 0.1, n = 3 (BRD4, BD1 selectivity = 500-fold); Screened at 10 µM against 45 GPCRs (PDSP screen), in-vitro potency of closest target: Ki(DRD4) = 1013.12 nM

In-vitro potencies of closest targets in BROMOScan (DiscoverX): pKd(TAF1, BD2) = 7.3, pKd(TAF1L, BD2) = 6.4, selective for the BD1 domain of BET proteins; Follow-up of closest targets (FRET assay): pIC50(TAF1, BD1) = 5.0 ± 0.1, n = 3 (BRD4, BD1 selectivity = 500-fold); Screened at 10 µM against 45 GPCRs (PDSP screen), in-vitro potency of closest target: Ki(DRD4) = 1013.12 nM

In-vitro potencies of closest targets in BROMOScan (DiscoverX): pKd(TAF1, BD2) = 7.3, pKd(TAF1L, BD2) = 6.4, selective for the BD1 domain of BET proteins; Follow-up of closest targets (FRET assay): pIC50(TAF1, BD1) = 5.0 ± 0.1, n = 3 (BRD4, BD1 selectivity = 500-fold); Screened at 10 µM against 45 GPCRs (PDSP screen), in-vitro potency of closest target: Ki(DRD4) = 1013.12 nM

In-vitro potencies of closest targets in BROMOScan (DiscoverX): pKd(TAF1, BD2) = 5.4, pKd(EP300) = 5.1, pKd(CREBBP) = 5.0, pKd(EP300) = 5.1; Cross screening in liability panel against 48 targets (GPCRs, transporter, ion-channels, and other targets), closest target: pEC50(CB2 receptor) <5; Selective for BD2 domain of BET proteins; Screened at 10 µM against 45 GPCRs (PDSP screen), closest target as % inhibition: OPRK1(40%);

In-vitro potencies of closest targets in BROMOScan (DiscoverX): pKd(TAF1, BD2) = 5.4, pKd(EP300) = 5.1, pKd(CREBBP) = 5.0, pKd(EP300) = 5.1; Cross screening in liability panel against 48 targets (GPCRs, transporter, ion-channels, and other targets), closest target: pEC50(CB2 receptor) <5; Selective for BD2 domain of BET proteins; Screened at 10 µM against 45 GPCRs (PDSP screen), closest target as % inhibition: OPRK1(40%);

In-vitro potencies of closest targets in BROMOScan (DiscoverX): pKd(TAF1, BD2) = 5.4, pKd(EP300) = 5.1, pKd(CREBBP) = 5.0, pKd(EP300) = 5.1; Cross screening in liability panel against 48 targets (GPCRs, transporter, ion-channels, and other targets), closest target: pEC50(CB2 receptor) <5; Selective for BD2 domain of BET proteins; Screened at 10 µM against 45 GPCRs (PDSP screen), closest target as % inhibition: OPRK1(40%);

In-vitro potencies of closest targets in BROMOScan (DiscoverX): pKd(TAF1, BD2) = 5.4, pKd(EP300) = 5.1, pKd(CREBBP) = 5.0, pKd(EP300) = 5.1; Cross screening in liability panel against 48 targets (GPCRs, transporter, ion-channels, and other targets), closest target: pEC50(CB2 receptor) <5; Selective for BD2 domain of BET proteins; Screened at 10 µM against 45 GPCRs (PDSP screen), closest target as % inhibition: OPRK1(40%);

Screened at 20 µM, dTm(SMARCA2) = 7.7 K, dTm(SMARCA4) = 7.4 K, dTm(PBRM1,BD5) = 11.3 K, other targets <2.7 K; Screened at 20 µM against 85 protein kinases (DSF assay), clean selectivity profile with all targets dTm <0.5 K

Screened at 20 µM, dTm(SMARCA2) = 7.7 K, dTm(SMARCA4) = 7.4 K, dTm(PBRM1,BD5) = 11.3 K, other targets <2.7 K; Screened at 20 µM against 85 protein kinases (DSF assay), clean selectivity profile with all targets dTm <0.5 K

Screened at 20 µM, dTm(SMARCA2) = 7.7 K, dTm(SMARCA4) = 7.4 K, dTm(PBRM1,BD5) = 11.3 K, other targets <2.7 K; Screened at 20 µM against 85 protein kinases (DSF assay), clean selectivity profile with all targets dTm <0.5 K

Screened at 10 µM against 45 GPCRs (PDSP screen), in-vitro potency of closest targets: Ki(HTR7) >10 µM, Ki(GABA/PBR) >10 µM, Ki(HTR1D) = 6453.57 nM; Selectivity within target family (GLS2 enzyme activity assay): IC50 >30 µM (> 1000 fold); Selectivity outside target family (probe selectivity observed in Proteome Integral Stability Assay): closest targets in CEREP binding assay at 10 μM (percent inhibition of control specific binding): ADORA3 (agonist radioligand) = 76.6%, CHRM1 (antagonist radioligand) = 92.6%, CHRM2 (antagonist radioligand) = 71.5%

Screened at 10 µM, in-vitro potencies of closest targets: Ki(TMEM97) = 1476.05 nM, Ki(SIGMAR1) = 1828.52 nM, Ki(GABA/PBR) = 1805.52 nM, Ki(OPRK1) = 3005.38 nM; Screened in enzymatic assays against other CCR family members: inactive on hCCR3, IC50(hCCR2) >30 µM, IC50(hCCR4) >30 µM, IC50(hCCR%) >30 µM, IC50(hCCR6) >30 µM, IC50(hCCR7) >30 µM, IC50(hCCR8) >30 µM, IC50(hCCR9) >30 µM, IC50(hCCR10) >30 µM, IC50(hCXCR1) >30 µM, IC50(hCXCR2) >30 µM, IC50(hCXCR3) >30 µM, IC50(hCXCR4) >30 µM, IC50(hCXCR5) >30 µM

Screened at 1 µM in quantitative proteomic studies, clean selectivty profile

Screened at 1 µM, 10 µM, and 50 µM (methyltransferase activity of G9a, GLP, SUV39H1, SUV39H2, SETDB1, SETD8, SUV420H1, SUV420H2, SETD7, MLL1 pentameric complex, MLL3 pentameric complex, EZH2 trimeric complex, PRMT1, PRMT3, PRMT4, PRMT5-MEP50 complex, PRMT6, PRMT7, PRMT8, PRMT9, PRDM9, SETD2, SMYD2, SMYD3, BCDIN3D, METTL3-14, and DNMT1), closest targets SUV420H1 (60% activity at 1 µM), SUV420H2 (62% activity at 1 µM); In-cellular follow-up by western blot assay (effect on H4K20me2/3 cellular mark modulated by SUVH420H1/H2 using MCF7 cells): EC50(SUVH420H1) >50 µM, EC50(SUV420H2) >50 µM; Screened at 1 µM against 40 kinases (enzymatic assay), closest target as % of inhibition: MAP4K4 (10.42%), clean selectivity profile; Screened against 15 diverse protein targets, all targets >10 µM, clean selectivity profile

DSF screen: Selective for NSD2-PWWP1 over 15 other human PWWP domains; clean panel of 33 methyltransferase domains, including the H3K36 methyltransferases NSD1, NSD2, NSD3 and SETD2;

Screened at 10 µM, in-vitro potencies of closest targets: Ki(HTR3A) = 584.83 nM, Ki(TMEM97) = 1099.22 nM, Ki(ADRA2A) = 8468.01 nM

Screened at 10 µM, in-vitro potency of closest target: Ki(HRH2) = 9781.38 nM; Selectivity against family members in biochemical assays, all >30 fold: IC50(hTRPV1) >25 µM, IC50(hTRPV4) >25 µM, IC50(hTRPC3) n.d., IC50(hTRPC5) = 10.0 µM, IC50(hTRPC6) >30 µM, IC50(hKCNK9 (TASK-3)) n.d., IC50(hCACNA1H (Cav3.2)) n.d.

Screened at 10 µM, in-vitro potency of closest target: KI(GABA/BZP) = 2539.22 nM

Screened at 10 µM, in-vitro potency of closest target: KI(GABA/BZP) = 2539.22 nM

Screened at 10 µM, in-vitro potency of closest target: KI(GABA/BZP) = 2539.22 nM

Screened at 10 µM, in-vitro potency of closest target: KI(GABA/BZP) = 2539.22 nM

Screened at 10 µM, in-vitro potency of closest target: Ki(GABA/BZP) = 473.91 nM

Screened at 10 µM, in-vitro potency of closest target: Ki(GABA/BZP) = 473.91 nM

Screened at 10 µM, in-vitro potency of closest target: Ki(GABA/BZP) = 473.91 nM

Screened at 10 µM, in-vitro potency of closest target: Ki(GABA/BZP) = 473.91 nM

rem. activity(RORA) = 0.56, rem. activity(RORB) = 0.53, rem. activity(RORC) = 0.46, rel. activity(RXRA)=30.83% to bexarotene 1 µM, rem. activity(NR2E1) = 0.42, rem. activity(NR5A1) = 0.42, inactive at 1 µM for UL48, HNF4A, NR2C1, NR4A1

rem. activity(RORA) = 0.56, rem. activity(RORB) = 0.53, rem. activity(RORC) = 0.46, rel. activity(RXRA)=30.83% to bexarotene 1 µM, rem. activity(NR2E1) = 0.42, rem. activity(NR5A1) = 0.42, inactive at 1 µM for UL48, HNF4A, NR2C1, NR4A1

rem. activity(RORA) = 0.56, rem. activity(RORB) = 0.53, rem. activity(RORC) = 0.46, rel. activity(RXRA)=30.83% to bexarotene 1 µM, rem. activity(NR2E1) = 0.42, rem. activity(NR5A1) = 0.42, inactive at 1 µM for UL48, HNF4A, NR2C1, NR4A1

fold-activation(NR5A1)=2.0, inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1

fold-activation(NR5A1)=2.0, inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1

fold-activation(PPARD) = 6.61, inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

fold-activation(PPARD) = 6.61, inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

fold-activation(PPARG) = 2.82, inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

fold-activation(PPARG) = 2.82, inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

fold-activation(PPARD) = 19.95, inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

fold-activation(PPARD) = 19.95, inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

fold-activation(PPARA) = 5.01, inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

fold-activation(PPARA) = 5.01, inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

rem. activity(PPARG)= 0.83, inactive at 1 µM for PPARA, UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

rem. activity(NR2C1) = 0.46, inactive at 10 µM for UL48, RXRA, HNF4A, NR2E1, NR4A1, NR5A1

rem. activity(NR2C1) = 0.46, inactive at 10 µM for UL48, RXRA, HNF4A, NR2E1, NR4A1, NR5A1

fold-activation(NR4A1) = 2.05, inactive at 10 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR5A1

rem. activity(NR2C1) = 0.49, rem. activity(NR2E1) = 0.48, inactive at 10 µM for UL48, RXRA, HNF4A, NR4A1, NR5A1

rem. activity(NR2C1) = 0.49, rem. activity(NR2E1) = 0.48, inactive at 10 µM for UL48, RXRA, HNF4A, NR4A1, NR5A1

no significance to UL48-effect, inactive at 1 µM for RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

inactive at 10 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

inactive at 10 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

inactive at 10 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1