Chemogenomic set

1.2 fold activation for VDR, active at 1 µM for VP16, inacitve at 1 µM for NR1I3 and RXRA

Screened at 1 µM against NR1I1, NR1I3, NR2A1, NR2B1, NR2C1, NR2E1, NR4A1, NR5A1 and VP16 in a Gal4 hybrid reporter gene assay (HEK293T cells were cotransfected with pFR-Luc reporter, pRL-SV40 control reporter and pFA-CMV containing the Gal4-DBD fused with human NR-LBD of interest or pECE-SV40-Gal4-VP16): clean selectivty profile with no significant agonism/antagonism detected; Screened at 20 µM in a bromodomain and kinase panel against ABL1, AURKA, BRD4, BRPF1, CDK2, CSNK1D, FGFR3, GSK3B, MAPK1, TRIM24 in DSF assays: clean selectivity profile with no significant binding (?Tm < 2 K)

Screened at 1 µM against NR1A1, NR1B1, NR1C3, NR1F3, NR1H3, NR1I1, NR1I2, NR1I3, NR2A1, NR2B1, NR4A1, NR5A2 and VP16 in a Gal4 hybrid reporter gene assay (HEK293T cells were cotransfected with pFR-Luc reporter, pRL-SV40 control reporter and pFA-CMV containing the Gal4-DBD fused with human NR-LBD of interest or pECE-SV40-Gal4-VP16): clean selectivty profile with no significant agonism/antagonism detected; Screened at 20 µM in a bromodomain and kinase panel against ABL1, AURKA, BRD4, BRPF1, CDK2, CSNK1D, FGFR3, GSK3B, MAPK1, TRIM24 in DSF assays: clean selectivity profile with no significant binding (?Tm < 2 K)

Screened at 1 µM againstNR1A1, NR1B1, NR1C3, NR1F3, NR1H3, NR1I1, NR1I2, NR1I3, NR2A1, NR2B1, NR4A1, NR5A2 and VP16 in a Gal4 hybrid reporter gene assay (HEK293T cells were cotransfected with pFR-Luc reporter, pRL-SV40 control reporter and pFA-CMV containing the Gal4-DBD fused with human NR-LBD of interest or pECE-SV40-Gal4-VP16): clean selectivty profile with no significant agonism/antagonism detected; Screened at 20 µM in a bromodomain and kinase panel against ABL1, AURKA, BRD4, BRPF1, CDK2, CSNK1D, FGFR3, GSK3B, MAPK1, TRIM24 in DSF assays: clean selectivity profile with no significant binding (?Tm < 2 K)

fold-activation(PPARG) = 3.39, inactive at 1 µM for PPARD, UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

Screened at 1 µM against NR1C2, NR1C3, NR2A1, NR2B1, NR2C1, NR2E1, NR4A1, NR5A1 and VP16 in a Gal4 hybrid reporter gene assay (HEK293T cells were cotransfected with pFR-Luc reporter, pRL-SV40 control reporter and pFA-CMV containing the Gal4-DBD fused with human NR-LBD of interest or pECE-SV40-Gal4-VP16): clean selectivty profile with no significant agonism/antagonism detected; Screened at 20 µM in a bromodomain and kinase panel against ABL1, AURKA, BRD4, BRPF1, CDK2, CSNK1D, FGFR3, GSK3B, MAPK1, TRIM24 in DSF assays: clean selectivity profile with no significant binding (?Tm < 2 K)

Screened at 1 µM against NR1I2, NR1I3, NR2A1, NR2B1, NR2C1, NR2E1, NR4A1, NR5A1 and VP16 in a Gal4 hybrid reporter gene assay (HEK293T cells were cotransfected with pFR-Luc reporter, pRL-SV40 control reporter and pFA-CMV containing the Gal4-DBD fused with human NR-LBD of interest or pECE-SV40-Gal4-VP16): clean selectivty profile with no significant agonism/antagonism detected; Screened at 20 µM in a bromodomain and kinase panel against ABL1, AURKA, BRD4, BRPF1, CDK2, CSNK1D, FGFR3, GSK3B, MAPK1, TRIM24 in DSF assays: clean selectivity profile with no significant binding (?Tm < 2 K)

inactive at 1 µM for NR1I2, NR1I3, UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

inactive at 1 µM for NR1I2, NR1I3, UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

Screened at 1 µM against NR1I2, NR1I3, NR2A1, NR2B1, NR2C1, NR2E1, NR4A1, NR5A1 and VP16 in a Gal4 hybrid reporter gene assay (HEK293T cells were cotransfected with pFR-Luc reporter, pRL-SV40 control reporter and pFA-CMV containing the Gal4-DBD fused with human NR-LBD of interest or pECE-SV40-Gal4-VP16): clean selectivty profile with no significant agonism/antagonism detected; Screened at 20 µM in a bromodomain and kinase panel against ABL1, AURKA, BRD4, BRPF1, CDK2, CSNK1D, FGFR3, GSK3B, MAPK1, TRIM24 in DSF assays: clean selectivity profile with no significant binding (?Tm < 2 K)

Screened at 1 µM againstNR1A1, NR1B1, NR1C3, NR1F3, NR1H3, NR1I1, NR1I2, NR1I3, NR2A1, NR2B1, NR4A1, NR5A2 and VP16 in a Gal4 hybrid reporter gene assay (HEK293T cells were cotransfected with pFR-Luc reporter, pRL-SV40 control reporter and pFA-CMV containing the Gal4-DBD fused with human NR-LBD of interest or pECE-SV40-Gal4-VP16): clean selectivty profile with no significant agonism/antagonism detected; Screened at 20 µM in a bromodomain and kinase panel against ABL1, AURKA, BRD4, BRPF1, CDK2, CSNK1D, FGFR3, GSK3B, MAPK1, TRIM24 in DSF assays: clean selectivity profile with no significant binding (?Tm < 2 K)

Screened at 1 µM againstNR1A1, NR1B1, NR1C3, NR1F3, NR1H3, NR1I1, NR1I2, NR1I3, NR2A1, NR2B1, NR4A1, NR5A2 and VP16 in a Gal4 hybrid reporter gene assay (HEK293T cells were cotransfected with pFR-Luc reporter, pRL-SV40 control reporter and pFA-CMV containing the Gal4-DBD fused with human NR-LBD of interest or pECE-SV40-Gal4-VP16): clean selectivty profile with no significant agonism/antagonism detected; Screened at 20 µM in a bromodomain and kinase panel against ABL1, AURKA, BRD4, BRPF1, CDK2, CSNK1D, FGFR3, GSK3B, MAPK1, TRIM24 in DSF assays: clean selectivity profile with no significant binding (?Tm < 2 K)

Screened at 1 µM against NR2A1, NR2B1, NR2C1, NR2E1, NR4A1, NR5A1 and VP16 in a Gal4 hybrid reporter gene assay (HEK293T cells were cotransfected with pFR-Luc reporter, pRL-SV40 control reporter and pFA-CMV containing the Gal4-DBD fused with human NR-LBD of interest or pECE-SV40-Gal4-VP16): clean selectivty profile with no significant agonism/antagonism detected; Screened at 20 µM in a bromodomain and kinase panel against ABL1, AURKA, BRD4, BRPF1, CDK2, CSNK1D, FGFR3, GSK3B, MAPK1, TRIM24 in DSF assays: clean selectivity profile with no significant binding (?Tm < 2 K)

Screened at 1 µM against NR2A1, NR2B1, NR2C1, NR2E1, NR4A1, NR5A1 and VP16 in a Gal4 hybrid reporter gene assay (HEK293T cells were cotransfected with pFR-Luc reporter, pRL-SV40 control reporter and pFA-CMV containing the Gal4-DBD fused with human NR-LBD of interest or pECE-SV40-Gal4-VP16): clean selectivty profile with no significant agonism/antagonism detected; Screened at 20 µM in a bromodomain and kinase panel against ABL1, AURKA, BRD4, BRPF1, CDK2, CSNK1D, FGFR3, GSK3B, MAPK1, TRIM24 in DSF assays: clean selectivity profile with no significant binding (?Tm < 2 K)

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

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

In vitro follow up of closest targets: Kd(IKKB) = 170 nM, Kd(YSK4) = 200 nM, Kd(CDK11A) = 399 nM, Kd(CDK11B) = 420 nM, Kd(CDK13) = 800 nM, Kd(CDK7) = 680 nM, Kd(MYLK4) = 700 nM; S(300nM) = 0.0052, S(3µM) = 0.0440;

Closest targets: Kd(GAK) = 79 nM, Kd(BLK) = 220 nM, Kd(ABL,F317L) = 230 nM, Kd(IRAK1) = 240 nM, KD(EPHA6) = 340 nM, Kd(HIPK4) = 360 nM, Kd(ABL,Q252H) = 380 nM, Kd(ABL,E255K) = 420 nM, Kd(PHKG2) = 470 nM,Kd(ABL,H396P) = 500 nM, Kd(ABL,phos.) = 570 nM, Kd(LCK) = 570 nM, Kd(ABL,F317L, nonphos.) = 580 nM, Kd(ABL,F317I, phos.) = 750 nM, Kd(ABL,Q252H, nonphos.) = 790 nM, Kd(ABL,Y253F, phos.) = 830 nM, Kd(ABL,T315I, phos.) = 870 nM, Kd(DYRK1A) = 970 nM, others >1 µM; Screened also against 52 kinases, closest targets: IC50(LYN) = 736 nM, IC50(ABL,T315I)= 1180 nM, others >2 µM (PMID: 18408761)

Closest targets: Kd(GAK) = 79 nM, Kd(BLK) = 220 nM, Kd(ABL,F317L) = 230 nM, Kd(IRAK1) = 240 nM, KD(EPHA6) = 340 nM, Kd(HIPK4) = 360 nM, Kd(ABL,Q252H) = 380 nM, Kd(ABL,E255K) = 420 nM, Kd(PHKG2) = 470 nM,Kd(ABL,H396P) = 500 nM, Kd(ABL,phos.) = 570 nM, Kd(LCK) = 570 nM, Kd(ABL,F317L, nonphos.) = 580 nM, Kd(ABL,F317I, phos.) = 750 nM, Kd(ABL,Q252H, nonphos.) = 790 nM, Kd(ABL,Y253F, phos.) = 830 nM, Kd(ABL,T315I, phos.) = 870 nM, Kd(DYRK1A) = 970 nM, others >1 µM; Screened also against 52 kinases, closest targets: IC50(LYN) = 736 nM, IC50(ABL,T315I)= 1180 nM, others >2 µM (PMID: 18408761)

Closest targets: Kd(GAK) = 79 nM, Kd(BLK) = 220 nM, Kd(ABL,F317L) = 230 nM, Kd(IRAK1) = 240 nM, KD(EPHA6) = 340 nM, Kd(HIPK4) = 360 nM, Kd(ABL,Q252H) = 380 nM, Kd(ABL,E255K) = 420 nM, Kd(PHKG2) = 470 nM,Kd(ABL,H396P) = 500 nM, Kd(ABL,phos.) = 570 nM, Kd(LCK) = 570 nM, Kd(ABL,F317L, nonphos.) = 580 nM, Kd(ABL,F317I, phos.) = 750 nM, Kd(ABL,Q252H, nonphos.) = 790 nM, Kd(ABL,Y253F, phos.) = 830 nM, Kd(ABL,T315I, phos.) = 870 nM, Kd(DYRK1A) = 970 nM, others >1 µM; Screened also against 52 kinases, closest targets: IC50(LYN) = 736 nM, IC50(ABL,T315I)= 1180 nM, others >2 µM (PMID: 18408761)

Closest targets with Kd <100 nM: NTRK2, targets with 100 nM < Kd < 1 µM: NTRK1, NTRK3, others >1 µM

S(Kd < 300 nM) =0.0026, S(Kd < 3µM) = 0.0078

S(Kd < 300 nM) =0.0026, S(Kd < 3µM) = 0.0078

Screened at 10 µM, closest targets as % inhibition: PDGFRA(V561D) (96%), RPS6KA6 (92%), RPS6KA3(92%), PKN2(92%), RPS6KA1(91%), PIM1 (90%); Screened against 48 kinases (GlaxoSmithKline panel), closest target: IC50(NEK2) = 158 nM;

Screened at 10 µM, closest targets as % of contr.: PLK1(4.3%); In vitro follow up of closest targets: IC50(GAK) = 140 nM, IC50(PLK1/3/4) = 61/ 1600/ 3100 nM, IC50(STK33) = 5000 nM

Screened at 10 µM, closest targets as % of contr.: PLK1(4.3%); In vitro follow up of closest targets: IC50(GAK) = 140 nM, IC50(PLK1/3/4) = 61/ 1600/ 3100 nM, IC50(STK33) = 5000 nM

Screened at 10 µM, closest targets as % of contr.: ALK(8.9%), CLK1(7.3%), ERK2(6%), GAK(5.4%), IGF1R(0.9%), INSR(0.15%), INSRR(2%), JNK1(5.2%), LTK(1.2%), MEK4(6.2%), PRK2(0.1%), PTK2B(0.5%), RIOK3(0.5%); In vitro follow up of closest targets: IC50(ALK) = 21 nM, IC50(CLK1) = 1900 nM, IC50(ERK2) = 2900 nM, IC50(FAK) = 440 nM, IC50(GAK) = 1100 nM, IC50(IGF1R) = 750 nM, IC50(INSR) = 470 nM, IC50(INSRR) = 1200 nM, IC50(LTK) = 29 nM, IC50(PYK2) = 280 nM, IC50(TNK1) = 2600 nM, IC50(TNK2) = 3100 nM;

Screened at 0.5 µM, closest targets: IC50(DAPK3) = 17 nM, IC50(FLT3) = 35 nM, IC50(TBK1) = 35 nM, IC50(CLK3) = 41 nM, IC50(HIPK3) = 45 nM, IC50(PIM1) = 46 nM, IC50(CDK1/cyclinB) = 56 nM (radiometric filter-binding assay, conc. ATP = Km); Follow-up in cellular assays: IC50(FLT3/ PIM1/ CDK1) >10 µM;

Screened at 1 µM, closest targets as % inhibition: MAP4K2 (99%), MET(86%); Screened also with ProfilerPro® Kinase Selectivity Assay Kit (Caliper Life Sciences) at 1 µM, 88 kinases <50% inhibition, clean in IC50 follow up assay: IC50 >10 µM (PMID: 21425998)

Screened at 1 µM, closest targets as % inhibition: MAP4K2 (99%), MET(86%); Screened also with ProfilerPro® Kinase Selectivity Assay Kit (Caliper Life Sciences) at 1 µM, 88 kinases <50% inhibition, clean in IC50 follow up assay: IC50 >10 µM (PMID: 21425998)

Screened at 1 µM, closest targets as % inhibition: MAP4K2 (99%), MET(86%); Screened also with ProfilerPro® Kinase Selectivity Assay Kit (Caliper Life Sciences) at 1 µM, 88 kinases <50% inhibition, clean in IC50 follow up assay: IC50 >10 µM (PMID: 21425998)

Screened at 1 µM and 10 µM, no target >50% inhibition; Screened in Dundee panel (51 kinases at 10 µM), no target >50% inhibition; Screened in Pfizer panel (48 kinases), no target >50% inhibition; Screened in Millipore panel (105 kinases at 1 µM an 10 µM), closest targets: ALK, EpHB2, INSR; IC50 follow-up showed all IC50 >10 µM; PF-04217903 is >1000-fold selective for c-Met;

Screened at 20 µM, closest targets with dTm >5K: dTm(STK6) = 8.23K, dTm(STK10) = 7.64 K, dTm(PLK4) = 5.33 K, further targets dTm 3-5K: dTm(BMX) = 4.14 K, dTm(ULK3) = 4.13 K, dTm(SLK) = 3.82 K, dTm(AURKB) = 3.81 K, dTm(ABL1) = 3.52 K; HotSpot assay (Reaction Biology), 30 targets, in-vitro potencies of closest targets: IC50(ABL1) = 705.9 nM, IC50(BTK) = 871.7 nM, IC50(LCK) = 432.3 nM, IC50(LYN) = 346.7 nM, https://pubmed.ncbi.nlm.nih.gov/22566699/;

Screened at 20 µM, closest targets with dTm >1K: dTm(MARK3) = 1.76 K, dTm(MST4) = 1.33 K, dTm(GSK3B) = 1.33 K; Screened against 88 targets in functional assays (AstraZeneca): >200-fold with exception of PIK3 isoforms, PIK3CA (86-fold) and PIK3CG (108-fold), as well as DNA-PK (42-fold), IC50(PIK3CA) = 0.87 µM (AlphaScreen), IC50(PIK3CG) = 1.09 µM (AlphaScreen), IC50(DNA-PK) = 0.42 µM (ELISA), https://pubmed.ncbi.nlm.nih.gov/22906130/;

Screened at 20 µM, closest targets with dTm >1K: dTm(MARK3) = 1.76 K, dTm(MST4) = 1.33 K, dTm(GSK3B) = 1.33 K; Screened against 88 targets in functional assays (AstraZeneca): >200-fold with exception of PIK3 isoforms, PIK3CA (86-fold) and PIK3CG (108-fold), as well as DNA-PK (42-fold), IC50(PIK3CA) = 0.87 µM (AlphaScreen), IC50(PIK3CG) = 1.09 µM (AlphaScreen), IC50(DNA-PK) = 0.42 µM (ELISA), https://pubmed.ncbi.nlm.nih.gov/22906130/;

Screened at 10 µM, Ambit score <10 at 10 µM, off-targets with PoC <50%

Screened at 1 µM, in-vitro potency of closest targets: IC50(ASK2)= 510 nM, IC50(MEKK1) >10 µM, IC50(TAK1) >10 µM, IC50(IKKB) >10 µM, IC50(ERK1) >10 µM, IC50(JNK1) >10µM, IC50(MAPK14) >10 µM, IC50(GSK3B) >10 µM, IC50(PRKCT) >10 µM; Screened against 12 kinases (Invitrogen, enzymatic assays), in-vitro potency of closest targets: IC50(CDK2) = 0.158 µM, IC50(EGFR) <20 µM, IC50(EPHA3) <20 µM, IC50(ERBB4) <20 µM, IC50(GSK3B) = 0.832 µM, IC50(ITK) = 5.5 µM, IC50(MER) = 6.5 µM, IC50(MAPK14) <20 µM, IC50(PRKAA1) = 13.8 µM, IC50(RET) = 11.5 µM, IC50(ROCK1) = 4.9 µM, IC50(RON) <20 µM, https://pubmed.ncbi.nlm.nih.gov/28291695/; Screened at 20 µM against 100 kinases, DSF panel, closest targets with dTm >5 K: dTm(MAP3K5) = 12.41 K, dTm(CLK1) = 5.46 K, closest targets with dTm 3-5 K: dTm(CAMKK2) = 4.55 K, dTm(RPS6KA5) = 3.13 K, in-house data SGC Frankfurt;

Screened at 10 µM (Ambit phage-based competition binding assay), closest targets as % of contr.: FRK (0.55%), FLT4 (0.85%), FLT3 (0.05%), EPHA3 (0.1%), CSF1R (0.8%), BLK (0.35%), STK10 (0%), AURKB (0%), KIT (0%), LCK (0.2%), MUSK (0%), PDGFRA/B (0.2/0.7%), TEK (0.15%), NTRK1/2/3 (0.55/ 0.1/ 0.1%), KDR (0.2%), MKNK2 (0.75%); In vitro follow up of closest targets: IC50(MERTK) = 14 nM, IC50(FLT3) = 16 nM, IC50(AURKB) = 78 nM, IC50(LCK) = 120 nM, IC50(KDR) = 180 nM, IC50(NTRK1/2) = 290/ 190 nM, IC50(PKA) = 550, IC50 >2 µM for: BTK, CDK2E, EGFR, GSK3B, IGFR1R, IKK1/2, INSR, JAK2, MAPKAPK2, MAPK14, PIM1, PLK1/3

Screened at 10 µM (Ambit phage-based competition binding assay), closest targets as % of contr.: FRK (0.55%), FLT4 (0.85%), FLT3 (0.05%), EPHA3 (0.1%), CSF1R (0.8%), BLK (0.35%), STK10 (0%), AURKB (0%), KIT (0%), LCK (0.2%), MUSK (0%), PDGFRA/B (0.2/0.7%), TEK (0.15%), NTRK1/2/3 (0.55/ 0.1/ 0.1%), KDR (0.2%), MKNK2 (0.75%); In vitro follow up of closest targets: IC50(MERTK) = 14 nM, IC50(FLT3) = 16 nM, IC50(AURKB) = 78 nM, IC50(LCK) = 120 nM, IC50(KDR) = 180 nM, IC50(NTRK1/2) = 290/ 190 nM, IC50(PKA) = 550, IC50 >2 µM for: BTK, CDK2E, EGFR, GSK3B, IGFR1R, IKK1/2, INSR, JAK2, MAPKAPK2, MAPK14, PIM1, PLK1/3

Screened at 10 µM (Ambit phage-based competition binding assay), closest targets as % of contr.: FRK (0.55%), FLT4 (0.85%), FLT3 (0.05%), EPHA3 (0.1%), CSF1R (0.8%), BLK (0.35%), STK10 (0%), AURKB (0%), KIT (0%), LCK (0.2%), MUSK (0%), PDGFRA/B (0.2/0.7%), TEK (0.15%), NTRK1/2/3 (0.55/ 0.1/ 0.1%), KDR (0.2%), MKNK2 (0.75%); In vitro follow up of closest targets: IC50(MERTK) = 14 nM, IC50(FLT3) = 16 nM, IC50(AURKB) = 78 nM, IC50(LCK) = 120 nM, IC50(KDR) = 180 nM, IC50(NTRK1/2) = 290/ 190 nM, IC50(PKA) = 550, IC50 >2 µM for: BTK, CDK2E, EGFR, GSK3B, IGFR1R, IKK1/2, INSR, JAK2, MAPKAPK2, MAPK14, PIM1, PLK1/3

Screened at 10 µM (Ambit phage-based competition binding assay), closest targets as % of contr.: FRK (0.55%), FLT4 (0.85%), FLT3 (0.05%), EPHA3 (0.1%), CSF1R (0.8%), BLK (0.35%), STK10 (0%), AURKB (0%), KIT (0%), LCK (0.2%), MUSK (0%), PDGFRA/B (0.2/0.7%), TEK (0.15%), NTRK1/2/3 (0.55/ 0.1/ 0.1%), KDR (0.2%), MKNK2 (0.75%); In vitro follow up of closest targets: IC50(MERTK) = 14 nM, IC50(FLT3) = 16 nM, IC50(AURKB) = 78 nM, IC50(LCK) = 120 nM, IC50(KDR) = 180 nM, IC50(NTRK1/2) = 290/ 190 nM, IC50(PKA) = 550, IC50 >2 µM for: BTK, CDK2E, EGFR, GSK3B, IGFR1R, IKK1/2, INSR, JAK2, MAPKAPK2, MAPK14, PIM1, PLK1/3

Screened at 1 µM, closest targets: IC50( FLT3) = 338 nM, IC50(FLT4) = 274 nM, IC50(CSF1R) = 488.9 nM, IC50(TYK2) = 116 nM

Screened at 1 µM, closest targets: IC50( FLT3) = 338 nM, IC50(FLT4) = 274 nM, IC50(CSF1R) = 488.9 nM, IC50(TYK2) = 116 nM

Screened at1 µM, closest targets as % of kinase activity.: STK33 (33%), ULK2 (37%)

Screened at1 µM, closest targets as % of kinase activity.: STK33 (33%), ULK2 (37%)

Screened at 0.1 µM, closest targets with PoC <50%, TAK1-TAB1

Screened at 1 µM, >1000-fold selective against other targets; Screened in Dundee panel against 140 kinases and 19 lipid kinases at 1 µM, >1000-fold selective; Screened in AstraZeneca panel against 8 lipid kinases, closest target: IC50(PIP5K1C) = 382 µM, IC50(PI4K3B) = 1.069 µM, IC50(PI3Kd) = 1.896 µM, others >2 µM; Screened in ProQuinase panel against 13 lipid kinases at 1 µM, closest targets: IC50(PIP5K1A) = 4.93 µM, IC50(PIP5K1C) = 3.37 µM;

Screened at 1 µM, closest targets as % of contr.: ABL (12%), BRK (13%), FYN (18%), KDR (17%)

Screened at 1 µM, closest targets as % of contr.: JNK2(4%), KIT(2%), PDGFRB(0%)

Screened at 10 µM, closest targets: IC50(MAPK15) = 130 nM, IC50(RPS6KA3) = 361 nM, IC50(RPS6KA2) = 362 nM; IC50(FLT3) = 582 nM, IC50(NUAK1) = 711 nM, IC50(CLK2) = 1370 nM (Z’-Lyte assay)

Screened at 10 µM, follow-up Kds and IC50s of closest targets: Kd(FLT1,D835Y) = 210 nM, Kd(CKL1) = 310 nM, Kd(MKNK2) = 330 nM, IC50(AKT3) = 427 nM.

Screened at 10 µM, follow-up Kds and IC50s of closest targets: Kd(FLT1,D835Y) = 210 nM, Kd(CKL1) = 310 nM, Kd(MKNK2) = 330 nM, IC50(AKT3) = 427 nM.

Screened at 1 µM, closest targets as % of contr.: ERBB2 (2.1%), ERBB4 (4.3%), TEC (6.4%); Inhibits kinases containing conserved cystein residue: IC50(BMX) = 46 nM, IC50(ERBB4) = 16 nM, IC50(TEC) = 93 nM, IC50(TXK) = 368 nM, IC50(ERBB2) = 1000 nM (Z’-LYTE assay), PMID: 26641137;

Screened at 200 nM, closest targets with >90% inhibition: IRAK1, targets with >70% inhibition: MNK2, LRRK2, CLK4, CK1g; Screened in KiNativ kinome screen (ActivX) in THP1 cell lysates at 10, 50, 200, 1000, and 5000 nM against 270 kinases; closest targets >50% inhibition at 200 nM: CSNK1G2, IRAK3, PIPK2C, CKSNK1D/E, https://pubmed.ncbi.nlm.nih.gov/28498658/; Cellular potency (NanoBRET assay in HEK293T cells): IC50(CSNK1G2) = 7.1 µM (SGC-Frankfurt, in-house data)

Screened at 1 µM, closest targets as % of contr.: BMX (15%), ROS1 (12%), TXK (14%), TNK2(10%)

Screened at 1 µM, closest targets as % of contr.: BMX (15%), ROS1 (12%), TXK (14%), TNK2(10%)

Screened at 1 µM, closest targets as % of contr.: BMX (15%), ROS1 (12%), TXK (14%), TNK2(10%)

Screened at 3 µM, closest targets as % inhibition: CLK1 (99%), DYRK1A/B (94/98%), DYRK3 (95%), PHKG (96%); In vitro follow-up (Invitrogen): IC50(DYRK3) = 99 nM, IC50(DYRK1A/B) = 104/157 nM, IC50(PHKG) = 136 nM, IC50(CLK3) = 300 nM; Cellular selectivity: IC50(JNK1/2/3) = 0.68 µM, IC50(CAMKK2) = 1.10 µM, IC50(PIP4K2C) = 1.30 µM (ActivX KiNative profiling assay (in HCT-116 cell line (CRL-1619))

Screened at 3 µM, closest targets as % inhibition: CLK1 (99%), DYRK1A/B (94/98%), DYRK3 (95%), PHKG (96%); In vitro follow-up (Invitrogen): IC50(DYRK3) = 99 nM, IC50(DYRK1A/B) = 104/157 nM, IC50(PHKG) = 136 nM, IC50(CLK3) = 300 nM; Cellular selectivity: IC50(JNK1/2/3) = 0.68 µM, IC50(CAMKK2) = 1.10 µM, IC50(PIP4K2C) = 1.30 µM (ActivX KiNative profiling assay (in HCT-116 cell line (CRL-1619))

Closest target: AKT3 (83% of ctrl.); Follow up in vitro activity: IC50(AKT3) = 618 nM (HTFR assay, PMID: 31584233), IC50(AKT3) = 4.3 µM (NanoBRET assay)

Closest target: AKT3 (83% of ctrl.); Follow up in vitro activity: IC50(AKT3) = 618 nM (HTFR assay, PMID: 31584233), IC50(AKT3) = 4.3 µM (NanoBRET assay)

Screened at 1 µM, 33P-ATP and filter-binding assay, closest target as % inhibition: BRK (99%), others >90%; In vitro potencies of closest 12 targets (33P-ATP and filter-binding assay (Reaction Biology)): IC50(ITK) = 56 nM, IC50(TEC) = 2.0 nM, IC50(JAK3) = 580 nM, IC50(EGFR) = 2.6 nM, IC50(HER2) = 530 nM, IC50(BRK) = 33 nM, IC50(FGR) = 155 nM, IC50(FRK/PTK5) = 379 nM, IC50(LCK) = 187 nM; Cellular selectivity: IC50(EGFR) = 606 nM (337-fold, p-EGFR HTRF assay), IC50(ITK) = 3477 nM (1932-fold, p-PLCg1 assay), IC50(TEC) = 204.7 nM (114-fold, p-TEC MSD assay), other targets not tested;

Screened at 1 µM, 33P-ATP and filter-binding assay, closest target as % inhibition: BRK (99%), others >90%; In vitro potencies of closest 12 targets (33P-ATP and filter-binding assay (Reaction Biology)): IC50(ITK) = 56 nM, IC50(TEC) = 2.0 nM, IC50(JAK3) = 580 nM, IC50(EGFR) = 2.6 nM, IC50(HER2) = 530 nM, IC50(BRK) = 33 nM, IC50(FGR) = 155 nM, IC50(FRK/PTK5) = 379 nM, IC50(LCK) = 187 nM; Cellular selectivity: IC50(EGFR) = 606 nM (337-fold, p-EGFR HTRF assay), IC50(ITK) = 3477 nM (1932-fold, p-PLCg1 assay), IC50(TEC) = 204.7 nM (114-fold, p-TEC MSD assay), other targets not tested;

Screened at 1 µM, 33P-ATP and filter-binding assay, closest target as % inhibition: BRK (99%), others >90%; In vitro potencies of closest 12 targets (33P-ATP and filter-binding assay (Reaction Biology)): IC50(ITK) = 56 nM, IC50(TEC) = 2.0 nM, IC50(JAK3) = 580 nM, IC50(EGFR) = 2.6 nM, IC50(HER2) = 530 nM, IC50(BRK) = 33 nM, IC50(FGR) = 155 nM, IC50(FRK/PTK5) = 379 nM, IC50(LCK) = 187 nM; Cellular selectivity: IC50(EGFR) = 606 nM (337-fold, p-EGFR HTRF assay), IC50(ITK) = 3477 nM (1932-fold, p-PLCg1 assay), IC50(TEC) = 204.7 nM (114-fold, p-TEC MSD assay), other targets not tested;

Screened at 1 µM, 33P-ATP and filter-binding assay, closest target as % inhibition: BRK (99%), others >90%; In vitro potencies of closest 12 targets (33P-ATP and filter-binding assay (Reaction Biology)): IC50(ITK) = 56 nM, IC50(TEC) = 2.0 nM, IC50(JAK3) = 580 nM, IC50(EGFR) = 2.6 nM, IC50(HER2) = 530 nM, IC50(BRK) = 33 nM, IC50(FGR) = 155 nM, IC50(FRK/PTK5) = 379 nM, IC50(LCK) = 187 nM; Cellular selectivity: IC50(EGFR) = 606 nM (337-fold, p-EGFR HTRF assay), IC50(ITK) = 3477 nM (1932-fold, p-PLCg1 assay), IC50(TEC) = 204.7 nM (114-fold, p-TEC MSD assay), other targets not tested;

Screened at 1 µM, 33P-ATP and filter-binding assay, closest target as % inhibition: BRK (99%), others >90%; In vitro potencies of closest 12 targets (33P-ATP and filter-binding assay (Reaction Biology)): IC50(ITK) = 56 nM, IC50(TEC) = 2.0 nM, IC50(JAK3) = 580 nM, IC50(EGFR) = 2.6 nM, IC50(HER2) = 530 nM, IC50(BRK) = 33 nM, IC50(FGR) = 155 nM, IC50(FRK/PTK5) = 379 nM, IC50(LCK) = 187 nM; Cellular selectivity: IC50(EGFR) = 606 nM (337-fold, p-EGFR HTRF assay), IC50(ITK) = 3477 nM (1932-fold, p-PLCg1 assay), IC50(TEC) = 204.7 nM (114-fold, p-TEC MSD assay), other targets not tested;

Tm panel screened at 20 µM, closest targets (dTm >5 K): dTm (CAMKK2) = 7.2 K, dTm(CLK1) = 6.4 K, dTm(EPHA2) = 6.2 K, dTm(NEK2) = 5.1 K, dTm(SRPK1) = 5.1 K, dTm(PHKG2) = 5.5 K, dTm(STK10) = 5.8 K, dTm(STK6) = 10.1 K, dTm(STK4) = 5.2 K, dTm(ULK3) = 5.8 K; Screened against 19 kinases (Radiometric inhibition assays): no target within 30-fold (PMID: 15323564)

Screened at 10 µM, closest target as % of contr.: PIK3CB (6.7%)

Screened at 10 µM, closest target as % of contr.: PIK3CB (6.7%)

Screened at 30 µM, highly specific

Screened at 30 µM, highly specific

Screened at 30 µM, highly specific

Screened at 30 µM, highly specific

In-vitro potencies of closest targets in the screen: Kd(MTOR) = 12 nM, Kd(HIPK3) = 310 nM, Kd(PIP5K2C) = 620 nM, Kd(RIOK2), Kd(DAPK3) = 840 nM, Kd(YSK4) = 930 nM, full screening data are available as supporting information;
Screened against 69 kinases at 1 µM, closest targets as % of control: SmMLCK (62%), HIPK2 (67%), full screening data available in paper, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2267365/;

In-vitro potencies of closest targets in the screen: Kd(MTOR) = 12 nM, Kd(HIPK3) = 310 nM, Kd(PIP5K2C) = 620 nM, Kd(RIOK2), Kd(DAPK3) = 840 nM, Kd(YSK4) = 930 nM, full screening data are available as supporting information;
Screened against 69 kinases at 1 µM, closest targets as % of control: SmMLCK (62%), HIPK2 (67%), full screening data available in paper, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2267365/;

In-vitro potencies of closest targets in the screen: Kd(MTOR) = 12 nM, Kd(HIPK3) = 310 nM, Kd(PIP5K2C) = 620 nM, Kd(RIOK2), Kd(DAPK3) = 840 nM, Kd(YSK4) = 930 nM, full screening data are available as supporting information;
Screened against 69 kinases at 1 µM, closest targets as % of control: SmMLCK (62%), HIPK2 (67%), full screening data available in paper, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2267365/;

In-vitro potencies of closest targets in the screen: Kd(MTOR) = 12 nM, Kd(HIPK3) = 310 nM, Kd(PIP5K2C) = 620 nM, Kd(RIOK2), Kd(DAPK3) = 840 nM, Kd(YSK4) = 930 nM, full screening data are available as supporting information;
Screened against 69 kinases at 1 µM, closest targets as % of control: SmMLCK (62%), HIPK2 (67%), full screening data available in paper, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2267365/;

In-vitro potencies of closest targets in the screen: Kd(MTOR) = 12 nM, Kd(HIPK3) = 310 nM, Kd(PIP5K2C) = 620 nM, Kd(RIOK2), Kd(DAPK3) = 840 nM, Kd(YSK4) = 930 nM, full screening data are available as supporting information;
Screened against 69 kinases at 1 µM, closest targets as % of control: SmMLCK (62%), HIPK2 (67%), full screening data available in paper, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2267365/;

In-vitro potencies of closest targets in the screen: Kd(MTOR) = 12 nM, Kd(HIPK3) = 310 nM, Kd(PIP5K2C) = 620 nM, Kd(RIOK2), Kd(DAPK3) = 840 nM, Kd(YSK4) = 930 nM, full screening data are available as supporting information;
Screened against 69 kinases at 1 µM, closest targets as % of control: SmMLCK (62%), HIPK2 (67%), full screening data available in paper, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2267365/;

Screened at 1 µM or 10 µM (iknase panel literature or Upstate/Millipore); Closest targets: Ki(VEGFR2) = 8 nM, IC50(YES) = 14 nM, IC50(FMS) = 10 nM, IC50(AURKA) = 23nM, IC50(FGFR3) = 23 nM, IC50(FLT3) = 25 nM, IC50(RET) = 39 nM;

Screened at 1 µM or 10 µM (Kinase panel literature or Upstate/Millipore); Closest targets: Ki(VEGFR2) = 8 nM, IC50(YES) = 14 nM, IC50(FMS) = 10 nM, IC50(AURKA) = 23nM, IC50(FGFR3) = 23 nM, IC50(FLT3) = 25 nM, IC50(RET) = 39 nM;

Screened at 10 µM, none target within 10-fold, closest target as % of contr.: PIK3CA (I800L, 33%)

Screened at 10 µM, none target within 10-fold, closest target as % of contr.: PIK3CA (I800L, 33%)

Kinases screened in Kinobeads assay,closest target: Kd(CAMKK2) = 195 nM; Kd (BRD4) = 79 nM.

Kinases screened in Kinobeads assay,closest target: Kd(CAMKK2) = 195 nM; Kd (BRD4) = 79 nM.

Kinases screened in Kinobeads assay,closest target: Kd(CAMKK2) = 195 nM; Kd (BRD4) = 79 nM.

In vitro activity of closest targets:IC50(FLT1/4) = 42/74 nM, IC50(TIE2) = 59 nM, IC50(SIK) = 70 nM, IC50(FGFR1) = 78 nM

In vitro activity of closest targets:IC50(FLT1/4) = 42/74 nM, IC50(TIE2) = 59 nM, IC50(SIK) = 70 nM, IC50(FGFR1) = 78 nM

Screened at 1 µM, closest targets as % of contr.: CDK9 (10%), FLT3(D835Y) (9%), PLK1 (3%); Screened additionally against 58 kinases (radiometric kinase assay), closesttargets: IC50(FLT3) = 510 nM, IC50(MELK) = 744 nM, IC50(CK2) = 826 nM, other targets IC50 >10 µM (PMID: 21470862)

Screened at 1 µM, closest targets: IC50(PRKD2) = 139 nM, IC50(PRKD3) = 219 nM, IC50(RET) = 161 nM, IC50(FLT3) = 714 nM, IC50(LRRK2) =16 nM, IC50(PRKD1) = 35 nM, IC50(CLK1/2) = 512/ 112 nM, IC50(FGFR2) = 320 nM, IC50(PDGFRA) = 956 nM; Screened against panel of 68 receptors, ion channels and protein targets, at 10 µM, closest targets: IC50(A1A) = 914 nM, IC50(A1B) = 52 nM, IC50(A1D) = NA, IC50(A2A) = 1420 nM, IC50(AB2) = 1820 nM, IC50(imidazoline I2) = 97 nM (PMID: 22128344)

In vitro inhibition (biochemical potency): >100 fold selective for MEK, other targets >10µM

Screened at 1 µM, closest targets as % of cont.: PIK3C2B (80.4%), PIK3CA (98.6%), PIK3CD (97.9%), PIK3CG (90.6%), PIK3C3 (69.9%); IC50 follow-up of closest targets: IC50(PIK3C2B) = 292 nM, IC50(PIK3C3) = 374 nM, >1000-fold selective

Screened at 1 µM, closest targets as % of cont.: PIK3C2B (80.4%), PIK3CA (98.6%), PIK3CD (97.9%), PIK3CG (90.6%), PIK3C3 (69.9%); IC50 follow-up of closest targets: IC50(PIK3C2B) = 292 nM, IC50(PIK3C3) = 374 nM, >1000-fold selective

Screened at 1 µM, closest targets as % of cont.: PIK3C2B (80.4%), PIK3CA (98.6%), PIK3CD (97.9%), PIK3CG (90.6%), PIK3C3 (69.9%); IC50 follow-up of closest targets: IC50(PIK3C2B) = 292 nM, IC50(PIK3C3) = 374 nM, >1000-fold selective

Screened at 1 µM, closest targets as % of cont.: PIK3C2B (80.4%), PIK3CA (98.6%), PIK3CD (97.9%), PIK3CG (90.6%), PIK3C3 (69.9%); IC50 follow-up of closest targets: IC50(PIK3C2B) = 292 nM, IC50(PIK3C3) = 374 nM, >1000-fold selective

Screened at 1 µM, closest target with PoC <10%: ABL2 (5.8%)