Chemogenomic set

Screened at 10 µM in DSF assays, dTm(BRD7) = 6.47 K, dTm(BRD9) = 9.21 K, closest target: dTm(CERC2) = 5.61 K, all other bromodomains <2 K; In vitro follow up of closest target: Kd(CERC2, ITC) = 200 nM; Screened against 32 bromodomains at 10 µM (Bromoscan and BromoMax

Screened at 10 µM in DSF assays, dTm(BRD7) = 6.47 K, dTm(BRD9) = 9.21 K, closest target: dTm(CERC2) = 5.61 K, all other bromodomains <2 K; In vitro follow up of closest target: Kd(CERC2, ITC) = 200 nM; Screened against 32 bromodomains at 10 µM (Bromoscan and BromoMax

Screened at 1 µM, in vitro activity of closest targets: Kd(CDK11) = 370 nM, Kd(EPHA8) = 550 nM, Kd(MUSK) = 530 nM

Screened at 1 µM, in vitro activity of closest targets: Kd(CDK11) = 370 nM, Kd(EPHA8) = 550 nM, Kd(MUSK) = 530 nM

Screened at 1 µM, in vitro activity of closest targets: Kd(CDK11) = 370 nM, Kd(EPHA8) = 550 nM, Kd(MUSK) = 530 nM

Screened at 1 µM, in vitro activity of closest targets: Kd(CDK11) = 370 nM, Kd(EPHA8) = 550 nM, Kd(MUSK) = 530 nM

Screened at 1 µM, in vitro activity of closest targets: Kd(CDK11) = 370 nM, Kd(EPHA8) = 550 nM, Kd(MUSK) = 530 nM

No targets with >70% inhibition at 100 nM and 500 nM

Screened at 1 µM, closest targets as % of activity: HIPK2 (99%), HIPK3 (94%), GSK3A (90%); Eurofins binding assay: IC50(HIPK1) = 187 nM, IC50(HIPK2) = 23 nM, IC50(CLK3) >3 µM; In-cellular potency (NanoBRET assay in HEK293T cells): Ki(HIPK2) >10 µM, Ki(HIPK4) = 5.41 µM

Screened at 1 µM, closest targets as % of activity: HIPK2 (99%), HIPK3 (94%), GSK3A (90%); Eurofins binding assay: IC50(HIPK1) = 187 nM, IC50(HIPK2) = 23 nM, IC50(CLK3) >3 µM; In-cellular potency (NanoBRET assay in HEK293T cells): Ki(HIPK2) >10 µM, Ki(HIPK4) = 5.41 µM

Screened at 1 µM, closest targets as % of activity: HIPK2 (99%), HIPK3 (94%), GSK3A (90%); Eurofins binding assay: IC50(HIPK1) = 187 nM, IC50(HIPK2) = 23 nM, IC50(CLK3) >3 µM; In-cellular potency (NanoBRET assay in HEK293T cells): Ki(HIPK2) >10 µM, Ki(HIPK4) = 5.41 µM

Screened at 1 µM, closest targets as % of activity: HIPK2 (99%), HIPK3 (94%), GSK3A (90%); Eurofins binding assay: IC50(HIPK1) = 187 nM, IC50(HIPK2) = 23 nM, IC50(CLK3) >3 µM; In-cellular potency (NanoBRET assay in HEK293T cells): Ki(HIPK2) >10 µM, Ki(HIPK4) = 5.41 µM

Screened at 1 µM, closest targets as % of activity: HIPK2 (99%), HIPK3 (94%), GSK3A (90%); Eurofins binding assay: IC50(HIPK1) = 187 nM, IC50(HIPK2) = 23 nM, IC50(CLK3) >3 µM; In-cellular potency (NanoBRET assay in HEK293T cells): Ki(HIPK2) >10 µM, Ki(HIPK4) = 5.41 µM

Screened at 1 µM, closest targets as % of activity: HIPK2 (99%), HIPK3 (94%), GSK3A (90%); Eurofins binding assay: IC50(HIPK1) = 187 nM, IC50(HIPK2) = 23 nM, IC50(CLK3) >3 µM; In-cellular potency (NanoBRET assay in HEK293T cells): Ki(HIPK2) >10 µM, Ki(HIPK4) = 5.41 µM

Screened at 1 µM, closest target as % of contr. PIK3CB (9.8%); Selective over PI3P, PI4P, PI34P, PI45P at 800 nM (radiolabel incorporation assay in NIH3T3 cells)

In vitro selectivity, screened at 1 µM, targets >90% of Ctrl. RET, LTK, PYK2,FLT3, ALK; KiNativ Technology screen against 265 kinases at 5 µM, Hela cells, highly selective after washout

In vitro selectivity, screened at 1 µM, targets >90% of Ctrl. RET, LTK, PYK2,FLT3, ALK; KiNativ Technology screen against 265 kinases at 5 µM, Hela cells, highly selective after washout

In vitro selectivity, screened at 1 µM, targets >90% of Ctrl. RET, LTK, PYK2,FLT3, ALK; KiNativ Technology screen against 265 kinases at 5 µM, Hela cells, highly selective after washout

Screened at 1 µM and 10 µM, highly selective within methyltransferase family; Selective over 9 reader proteins (UHRF1, BRPF1, WDR5, HGDF2, TDRD3, RBBP1, FXR1, EED, and SND1) in DSF assays: all enzymes dTm <2 K;

Screened at 1 µM and 10 µM, highly selective within methyltransferase family; Selective over 9 reader proteins (UHRF1, BRPF1, WDR5, HGDF2, TDRD3, RBBP1, FXR1, EED, and SND1) in DSF assays: all enzymes dTm <2 K;

Screened at 1 µM and 10 µM, highly selective within methyltransferase family; Selective over 9 reader proteins (UHRF1, BRPF1, WDR5, HGDF2, TDRD3, RBBP1, FXR1, EED, and SND1) in DSF assays: all enzymes dTm <2 K;

Screened at 1 µM and 10 µM, highly selective within methyltransferase family; Selective over 9 reader proteins (UHRF1, BRPF1, WDR5, HGDF2, TDRD3, RBBP1, FXR1, EED, and SND1) in DSF assays: all enzymes dTm <2 K;

Screened at 1 µM and 10 µM, highly selective within methyltransferase family; Selective over 9 reader proteins (UHRF1, BRPF1, WDR5, HGDF2, TDRD3, RBBP1, FXR1, EED, and SND1) in DSF assays: all enzymes dTm <2 K;

fold-activation(NR1H4) = 10.96, inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

fold-activation(NR1H4) = 10.96, inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

fold-activation(NR1H4) = 10.96, inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

fold-activation(NR1H4) = 10.96, inactive at 1 µM for UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

Screened at 1 µM, closest targets as % residual activity: RIPK2 (7%), VEGFR2 (11%), MINK1 (21%), VEGFR3 (28%), VEGFR1 (38%), MAP4K4 (41%), PDGFRA(V561D) (43%), PDGFRA (47%), TGFBR2 (47%); In-vitro follow-up of closest targets (radioisotopic protein kinase assay, 33PanQinase activity assay): IC50(RIPK2) = 54 nM, IC50(VEGFR2) = 97 nM (https://pubs.acs.org/doi/10.1021/jm500115w); Selectivity within ALK family (Radioisotopic protein kinase assay, HotSpot assay, Reaction Biology): IC50(ALK1) >10 µM, IC50(ALK2) >10 µM, IC50(ALK3) >10 µM, IC50(ALK4) = 0.013 µM, IC50(ALK5) = 0.011 µM, IC50(ALK6) >10 µM, IC50(ALK7) n.d. (https://pubs.acs.org/doi/10.1021/jm500115w);

Screened at 1 µM, closest targets as % residual activity: RIPK2 (7%), VEGFR2 (11%), MINK1 (21%), VEGFR3 (28%), VEGFR1 (38%), MAP4K4 (41%), PDGFRA(V561D) (43%), PDGFRA (47%), TGFBR2 (47%); In-vitro follow-up of closest targets (radioisotopic protein kinase assay, 33PanQinase activity assay): IC50(RIPK2) = 54 nM, IC50(VEGFR2) = 97 nM (https://pubs.acs.org/doi/10.1021/jm500115w); Selectivity within ALK family (Radioisotopic protein kinase assay, HotSpot assay, Reaction Biology): IC50(ALK1) >10 µM, IC50(ALK2) >10 µM, IC50(ALK3) >10 µM, IC50(ALK4) = 0.013 µM, IC50(ALK5) = 0.011 µM, IC50(ALK6) >10 µM, IC50(ALK7) n.d. (https://pubs.acs.org/doi/10.1021/jm500115w);

Screened at 1 µM, S(10) = 0.01, closest targets as % of contr.: JAK3 (6.6%), PIP3CG (2.8%), PIP5K2C (5.6%), SBK1 (4.6%);

Screened at 1 µM, S(10) = 0.01, closest targets as % of contr.: JAK3 (6.6%), PIP3CG (2.8%), PIP5K2C (5.6%), SBK1 (4.6%);

Screened at 10 µM, closest targets as % of contr.: CLK2 (25%), PIK3C2G (27%); Kd(mTOR) = 48 nM, Kd(PI3KC2b) = 40 nM (Kds by DiscoverX, PMID:28829592)

Screened at 10 µM, closest targets as % of contr.: CLK2 (25%), PIK3C2G (27%); Kd(mTOR) = 48 nM, Kd(PI3KC2b) = 40 nM (Kds by DiscoverX, PMID:28829592)

Screened at 10 µM, closest targets as % of contr.: CLK2 (25%), PIK3C2G (27%); Kd(mTOR) = 48 nM, Kd(PI3KC2b) = 40 nM (Kds by DiscoverX, PMID:28829592)

Screened at 10 µM, closest targets as % of contr.: CLK2 (25%), PIK3C2G (27%); Kd(mTOR) = 48 nM, Kd(PI3KC2b) = 40 nM (Kds by DiscoverX, PMID:28829592)

Screened at 10 µM, closest targets as % control activity: HIPK2 (47%)

Screened at 10 µM, closest targets as % control activity: HIPK2 (47%)

Screened at 10 µM, closest targets as % control activity: HIPK2 (47%)

Screened at 10 µM, closest targets as % control activity: HIPK2 (47%)

Screened at 1 µM, closest targets as % of control: TYK2 (20%), EGFR(L861Q, 24%), EGFR(L858R, 27%), full screening data available as supplement information;
Screened at 2 µM against 60 kinases, HotSpot assay platform (Reaction Biology), closest targets as % inhibition: SRC (87.6%), FLT3 (72.1%), LYN (77.4%), https://www.science.org/doi/10.1126/scitranslmed.3010277?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub%20%200pubmed;

Screened at 1 µM, closest targets as % of control: TYK2 (20%), EGFR(L861Q, 24%), EGFR(L858R, 27%), full screening data available as supplement information;
Screened at 2 µM against 60 kinases, HotSpot assay platform (Reaction Biology), closest targets as % inhibition: SRC (87.6%), FLT3 (72.1%), LYN (77.4%), https://www.science.org/doi/10.1126/scitranslmed.3010277?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub%20%200pubmed;

Screened at 1 µM, closest targets: IC50(AMPK) = 117 nM, IC50(BRSK1) = 187 nM, IC50(BRSK2) = 90 nM, IC50(CAMK2B) = 70 nM, IC50(DRAK1) = 71 nM, IC50(MNK2) = 148 nM, IC50(PHKG2) = 528 nM , IC50(PIM1) = 88 nM ,IC50(CDK1) = 354 nM, IC50(CDK5) = 902 nM, IC50(MAP3K5) = 60 nM, IC50(AXL) = 428 nM, IC50(MERTK) = 76 nM, IC50(RIPK2) = 590 nM;

Screened at 1 µM, closest targets: IC50(AMPK) = 117 nM, IC50(BRSK1) = 187 nM, IC50(BRSK2) = 90 nM, IC50(CAMK2B) = 70 nM, IC50(DRAK1) = 71 nM, IC50(MNK2) = 148 nM, IC50(PHKG2) = 528 nM , IC50(PIM1) = 88 nM ,IC50(CDK1) = 354 nM, IC50(CDK5) = 902 nM, IC50(MAP3K5) = 60 nM, IC50(AXL) = 428 nM, IC50(MERTK) = 76 nM, IC50(RIPK2) = 590 nM;

Screened at 1 µM, closest targets: IC50(AMPK) = 117 nM, IC50(BRSK1) = 187 nM, IC50(BRSK2) = 90 nM, IC50(CAMK2B) = 70 nM, IC50(DRAK1) = 71 nM, IC50(MNK2) = 148 nM, IC50(PHKG2) = 528 nM , IC50(PIM1) = 88 nM ,IC50(CDK1) = 354 nM, IC50(CDK5) = 902 nM, IC50(MAP3K5) = 60 nM, IC50(AXL) = 428 nM, IC50(MERTK) = 76 nM, IC50(RIPK2) = 590 nM;

Screened at 1 µM, 4h incubation, closest target as % of inhibition: CDK17 (94.2%), CDK18 (8.%), full screening data are available as supporting info,https://pubmed.ncbi.nlm.nih.gov/30930164/;

Screened at 1 µM, 4h incubation, closest target as % of inhibition: CDK17 (94.2%), CDK18 (8.%), full screening data are available as supporting info,https://pubmed.ncbi.nlm.nih.gov/30930164/;

Screened at 1 µM, 4h incubation, closest target as % of inhibition: CDK17 (94.2%), CDK18 (8.%), full screening data are available as supporting info,https://pubmed.ncbi.nlm.nih.gov/30930164/;

Screened at 1 µM, closest targets as % of inhibition: CDK9 (90.4%), CDK10 (82.6%), CDK5 (59.5%), CDK6(51.3%), CDK7 (54.7%), CHED (59.2%), PCTAIRE1 (89.5%), PCTAIRE2 (89.7%), PCTAIRE3 (91.1%), full screening data are available as supporting information, https://www.nature.com/articles/nchembio.2538;
In-vitro potency (Z'LYTE kinase assays, CDK1/CyclinB, conc. ATP = Km): IC50(CDK1) = 171 nM, https://www.nature.com/articles/nchembio.2538;
In-vitro potency (Z'LYTE kinase assays, CDK2/CyclinA, conc. ATP = Km): IC50(CDK2) = 62 nM, https://www.nature.com/articles/nchembio.2538;
In-vitro potency (Adapta Eu kinase assay, CDK7/CycH/MNAT1, conc. ATP = Km): IC50(CDK7) = 398 nM, https://www.nature.com/articles/nchembio.2538;
Quantitative mass-spectrometry-based proteomics: experiment identified CDK9 as the most depleted protein out of 4,512 proteins based on the quantification of two or more peptides, https://www.nature.com/articles/nchembio.2538

Screened at 10 µM, closest targets as % of control: BTK (1.4%), FER (6%), MAPK14 (11%), WNK1 (2.8%);

Screened at 20 µM, closest targets: PHKG2 = 3.0 K, BRD4 = 7.6 K, MSSK1 = 5.2 K, clean; Differential scanning fluorimetry screen against 12 related kinases, at 12.5 µM: ΔTm(SRPK1) = 12.8°C; ΔTm(SRPK2, CLK1) = 6.7 °C, ΔTm(CLK4) = 5.7 °C; other splicing kinases <2°CScreened at 1 µM against 50 kinases, Radiolabeled ATP competition assay (MRC Dundee Kinase Centre), closest targets: SRPK1 (96% inhibition), highly selective;

Screened at 20 µM, closest targets: PHKG2 = 3.0 K, BRD4 = 7.6 K, MSSK1 = 5.2 K, clean; Differential scanning fluorimetry screen against 12 related kinases, at 12.5 µM: ΔTm(SRPK1) = 12.8°C; ΔTm(SRPK2, CLK1) = 6.7 °C, ΔTm(CLK4) = 5.7 °C; other splicing kinases <2°CScreened at 1 µM against 50 kinases, Radiolabeled ATP competition assay (MRC Dundee Kinase Centre), closest targets: SRPK1 (96% inhibition), highly selective;

inactive at 1 µM for NR1H3, NR1H2, UL48, RXRA, HNF4A, NR2C1, NR2E1, NR4A1, NR5A1

fold-activation(NR1I2) = 2.24, fold-activation(NR2E1) = 1.84, inactive at 3 µM for NR1H4, UL48, RXRA, HNF4A, NR2C1, NR4A1, NR5A1

fold-activation(NR1I2) = 2.24, fold-activation(NR2E1) = 1.84, inactive at 3 µM for NR1H4, UL48, RXRA, HNF4A, NR2C1, NR4A1, NR5A1

Screened at 1 µM, closest targets AURKA (54%), PLK(53%), CLK1(43%), all IC50 >3.5 µM, In-vitro potency of closest target (Radiometric kinase assay ): Kd(CDK12) = 158 nM, https://www.nature.com/articles/nchembio.2166;

Screened at 100 µM in DSF assay, closest target: dTm(CEREBBP) = 1-3 K; Screened in CEREP panel against 50 GPCRs, ion channels and transporters at 10 µM, closest target as % inhibition: NK2 (48%)

Screened at 100 µM in DSF assay, closest target: dTm(CEREBBP) = 1-3 K; Screened in CEREP panel against 50 GPCRs, ion channels and transporters at 10 µM, closest target as % inhibition: NK2 (48%)

Screened at 100 µM in DSF assay, closest target: dTm(CEREBBP) = 1-3 K; Screened in CEREP panel against 50 GPCRs, ion channels and transporters at 10 µM, closest target as % inhibition: NK2 (48%)

Screened at 100 µM in DSF assay, closest target: dTm(CEREBBP) = 1-3 K; Screened in CEREP panel against 50 GPCRs, ion channels and transporters at 10 µM, closest target as % inhibition: NK2 (48%)

Screened at 10 µM and 50 µM: MS049 is highly selective and showed no inhibition of any enzyme in the panel; Screened in CEREP panel against 100 enzymes (GPCRs, ion channels, transporters, kinases) at 10 µM, clean profile; Screened against 45 GPCR targets (PDSP screen) at 10 µM, closest targets (radioligand binding assays): Ki(H3) = 87 nM, Ki(Sigma 1) = 64 nM, Ki(Sigma 2) = 574 nM;

Screened at 10 µM and 50 µM: MS049 is highly selective and showed no inhibition of any enzyme in the panel; Screened in CEREP panel against 100 enzymes (GPCRs, ion channels, transporters, kinases) at 10 µM, clean profile; Screened against 45 GPCR targets (PDSP screen) at 10 µM, closest targets (radioligand binding assays): Ki(H3) = 87 nM, Ki(Sigma 1) = 64 nM, Ki(Sigma 2) = 574 nM;

No inhibition of AP-1 and CRE (IC50 >100 µM, TOPFLASH reporter assay); no interaction with the beta-catenin transcriptional coactivator p300.

Screened at 10 µM, ATP-site competition binding assay, S(10) = 0.012, closest targets: Kd(BMK1) = 80 nM, Kd(DCAMKL2) = 190 nM, Kd(TNK1) = 890 nM, Kd(PLK4) = 600 nM; Cellular selectivity profiling using KiNativ method against 181 kinases in HeLa cells, closest targets: IC50(TNK1/2) = 10/ 18 µM, IC50(RSK1/2) = 29/ 47 µM, IC50(FAK) = 39 µM, IC50(PIK4CA/L) = 36 µM, others >50 µM;

Screened at 10 µM, closest targets as % of contr.: NTRK1 (59%); Screened against 300 proteins in Kinobeads assays, in-vitro potencies of closest targets: Kdapp(BRD4) = 1915 nM, Kdapp(MAP2K1) = 62 nM, Kdapp(MAP2K2) = 9 nM (https://www.science.org/doi/10.1126/science.aan4368)

Screened at 10 µM, closest targets as % of contr.: NTRK1 (59%); Screened against 300 proteins in Kinobeads assays, in-vitro potencies of closest targets: Kdapp(BRD4) = 1915 nM, Kdapp(MAP2K1) = 62 nM, Kdapp(MAP2K2) = 9 nM (https://www.science.org/doi/10.1126/science.aan4368)

Screened at 20 µM, closest targets with dTm >3K: dTm(AURKB) = 13.8 K, dTm(STK6) = 9.08 K, dTm(FLT1) = 4.33 K;

Screened at 10 µM, closest targets as % of control: CDKL2(3.8%), PLK1 (5.4%), PIK3CG (14%), TAOK1 (12%), TAOK2 (13%), full screening data available as supporting information, https://pubs.acs.org/doi/10.1021/acs.jmedchem.8b01040#; In-vitro potency (enzymatic assay, 22 kinases), all targets >5 µM, CDC2, ERK2, PRKCA, PRKCE, AKT1, RPS6KB1, RSK2, SRC, TEK, FLT1, KDR,bFGFRTK, IGF1RTK, INSR, AMPK, PDK1, CHEK1, CSNK1E, DNAPK, PIP3K; >800-fold selectivity against 23 additional enzymes and 22 receptors, https://pubmed.ncbi.nlm.nih.gov/12606497/; Screened at 1 µM (against 71 kinases, in vitro), closest targets as % of control PLK1 (41%), MELK(45%), CDK2/cyclinA(53%), full screening data available in paper, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2267365/;

Screened at 10 µM, closest targets as % of control: CDKL2(3.8%), PLK1 (5.4%), PIK3CG (14%), TAOK1 (12%), TAOK2 (13%), full screening data available as supporting information, https://pubs.acs.org/doi/10.1021/acs.jmedchem.8b01040#; In-vitro potency (enzymatic assay, 22 kinases), all targets >5 µM, CDC2, ERK2, PRKCA, PRKCE, AKT1, RPS6KB1, RSK2, SRC, TEK, FLT1, KDR,bFGFRTK, IGF1RTK, INSR, AMPK, PDK1, CHEK1, CSNK1E, DNAPK, PIP3K; >800-fold selectivity against 23 additional enzymes and 22 receptors, https://pubmed.ncbi.nlm.nih.gov/12606497/; Screened at 1 µM (against 71 kinases, in vitro), closest targets as % of control PLK1 (41%), MELK(45%), CDK2/cyclinA(53%), full screening data available in paper, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2267365/;

Kinases screened via competition binding assays: S(Kd < 300 nM) = 0.0026, S(Kd < 3µM) = 0.0052

Kinases screened via competition binding assays: S(Kd < 300 nM) = 0.0026, S(Kd < 3µM) = 0.0052

Screened at 1 µM, highly selective allosteric inhibitor

Screened at 1 µM, highly selective allosteric inhibitor

Screened at 1 µM, highly selective allosteric inhibitor

Screened at 1 µM, closest targets in the screen: DDR1, DDR2, KIT, ZAK, RSK, MYLK4, but confirmed by NanoBRET as false positive (IC50s of 910-140000 nM)

Screened at 1 µM, closest targets in the screen: DDR1, DDR2, KIT, ZAK, RSK, MYLK4, but confirmed by NanoBRET as false positive (IC50s of 910-140000 nM)

Screened at 1 µM, closest targets as % of inhibition: BLK (92%), CAMK2A (95%), EPHA2 (100%), EPHA4 (94%), FGFR2 (90%), FGR (97%), MAPKAPK3 (91%), NTRK3 (91%); Proteomics (Kinobeads assays), in-vitro potencies of closest targets: Kdapp(AURKA) = 5 nM, Kdapp(ACAD10) = 78 nM, Kdapp(AURKB) = 195 nM, Kdapp(PDPK1) = 289 nM, Kdapp(EPHA2) = 309 nM, Kdapp(EPHA4) = 455 nM, Kdapp(DNAJA1) = 962 nM, Kd(ABL2) = 1009 nM, Kdapp(GNAI2) = 184 nM, Kdapp(PSMA7) = 405 nM, Kdapp(MCM4) = 629 nM, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6542668/;

Screened at 10 µM, closest targets as % contr.: JAK3(JH1 catalytic domain) (3.2%), PRKACA (4.2%), PRKACB (6.4%); Additionally screened against 85 kinase targets at 1 µM, closest target as % of contr.: RPS6KA2 (20%), PMID: 19740074;

Screened at 10 µM, closest targets as % contr.: JAK3(JH1 catalytic domain) (3.2%), PRKACA (4.2%), PRKACB (6.4%); Additionally screened against 85 kinase targets at 1 µM, closest target as % of contr.: RPS6KA2 (20%), PMID: 19740074;

Screened at 1 µM, closest target as % of contr.: RAF1 (8%)

Screened at 0.1 µM, closest targets as % of contr.: CLK2 (0%), FLT3(N841I) (0%), FLT3(D835V) (0.2%), YSK4 (1.7%), JAK2 (4.8%), FLT3(D835Y) (5.9%), STK16 (7.7%), JAK3 (8.2), TTK (8.3%), FLT3(ITD,F691L) (9%), FLT3(D835H) (9.8%); Screened in Invitrogen panel at 10 µM, closest targets: IC50(NTRK1) = 56 nM, IC50(MAP3K9) = 75.3 nM; Screened in CEREP panel against 122 human receptors, ion channels, enzymes, closest target: Ki(ADORA3) = 27 nM; Screened in GPCR screen against 45 targets at 10 µM, clean profile;

Screened at 0.1 µM, closest targets as % of contr.: CLK2 (0%), FLT3(N841I) (0%), FLT3(D835V) (0.2%), YSK4 (1.7%), JAK2 (4.8%), FLT3(D835Y) (5.9%), STK16 (7.7%), JAK3 (8.2), TTK (8.3%), FLT3(ITD,F691L) (9%), FLT3(D835H) (9.8%); Screened in Invitrogen panel at 10 µM, closest targets: IC50(NTRK1) = 56 nM, IC50(MAP3K9) = 75.3 nM; Screened in CEREP panel against 122 human receptors, ion channels, enzymes, closest target: Ki(ADORA3) = 27 nM; Screened in GPCR screen against 45 targets at 10 µM, clean profile;

Closest targets: Kd(EPHA5) = 37.9 nM, other targets with low Kd values determined, however targets areof low- to no confidence; Cellular potency of closely related targets (NanoBRET assay in HEK293T cells): IC50(EPHA1) = 330 nM, IC50(EPHA3) = 1000 nM, IC50(EPHA4) = 100 nM, IC50(EPHA5) = 37 nM, IC50(EPHA6) = 2000 nM, IC50(EPHA7) = 2000 nM, IC50(EPHA8) = 12 nM, IC50(EPHB1) = 1000 nM, IC50(EPHB2) = 37 nM, IC50(EPHB3) = 330 nM, IC50(EPHB6) = 330 nM;Screened against 40 kinases in TR-FRET-based LanthaScreen™ and Caliper mobility shift assays, closest targets: IC50(RAF1) = 395 nM, IC50(c-Sarc) = 1266 nM, IC50(c-ABL) = 1667 nM, others >10 µM (PMID: 20803239)

Closest targets: Kd(EPHA5) = 37.9 nM, other targets with low Kd values determined, however targets areof low- to no confidence; Cellular potency of closely related targets (NanoBRET assay in HEK293T cells): IC50(EPHA1) = 330 nM, IC50(EPHA3) = 1000 nM, IC50(EPHA4) = 100 nM, IC50(EPHA5) = 37 nM, IC50(EPHA6) = 2000 nM, IC50(EPHA7) = 2000 nM, IC50(EPHA8) = 12 nM, IC50(EPHB1) = 1000 nM, IC50(EPHB2) = 37 nM, IC50(EPHB3) = 330 nM, IC50(EPHB6) = 330 nM;Screened against 40 kinases in TR-FRET-based LanthaScreen™ and Caliper mobility shift assays, closest targets: IC50(RAF1) = 395 nM, IC50(c-Sarc) = 1266 nM, IC50(c-ABL) = 1667 nM, others >10 µM (PMID: 20803239)

Screened at 1 µM, closest targets with <10% of contr.: ABL1, BLK, DDR1, FLT3, KIT, LCK, STK10, MAP2K5, MET, HIPK4, AURKB/C, FER, HCK, MST1R, CDK19, FRK, SRC, TIE1, CHEK2, MUSK, CDK8, CSF1R, EPHB6, PDGFRB, ROS1, NRTK1/3, MAP4K5, and mutants; Cellular selectivity (quantitative chemical proteomics, cell-based phosphorylation and proliferation assays in Hs578t cells), closest targets: Kd(cMET) = 13 nM,Kd(FER) = 107 nM, Kd(MAP2K5) = 124 nM, Kd(ABL1) = 154 nM, Kd(SRC) = 217 nM;

Screened at 1 µM, closest targets with <10% of contr.: ABL1, BLK, DDR1, FLT3, KIT, LCK, STK10, MAP2K5, MET, HIPK4, AURKB/C, FER, HCK, MST1R, CDK19, FRK, SRC, TIE1, CHEK2, MUSK, CDK8, CSF1R, EPHB6, PDGFRB, ROS1, NRTK1/3, MAP4K5, and mutants; Cellular selectivity (quantitative chemical proteomics, cell-based phosphorylation and proliferation assays in Hs578t cells), closest targets: Kd(cMET) = 13 nM,Kd(FER) = 107 nM, Kd(MAP2K5) = 124 nM, Kd(ABL1) = 154 nM, Kd(SRC) = 217 nM;

Screened at 1 µM, closest targets with <10% of contr.: ABL1, BLK, DDR1, FLT3, KIT, LCK, STK10, MAP2K5, MET, HIPK4, AURKB/C, FER, HCK, MST1R, CDK19, FRK, SRC, TIE1, CHEK2, MUSK, CDK8, CSF1R, EPHB6, PDGFRB, ROS1, NRTK1/3, MAP4K5, and mutants; Cellular selectivity (quantitative chemical proteomics, cell-based phosphorylation and proliferation assays in Hs578t cells), closest targets: Kd(cMET) = 13 nM,Kd(FER) = 107 nM, Kd(MAP2K5) = 124 nM, Kd(ABL1) = 154 nM, Kd(SRC) = 217 nM;

Screened at 10 µM, closest targets as % of contr.: SGK2 (0%), PRKX (4%), RSK4 (12%), SGK (13%), RSK3 (14%), NUAK1 (16%), ROCK2 (17%); Screened against panel of 13 kinases (PDK, AKT1, ALK5, ASK1, AURA, AURB, EGFR, GSK3B, IKK1, PIK3CG, ROCK1, SYK, VEGFR2): IC50(ODK) = 2.5 nM, IC50(AURB) = 3162 nM, IC50(ROCK1) = 7943 nM, others >10 µM;

Screened at 1 µM, follow-up Kds for closest targets: Kd(GSK3B) = 8.3 nM, Kd(PRKCD) = 25 nM, Kd(RSK4) = 25 nM, Kd(PRKCQ) = 36 nM, Kd(FLT3(D835Y)) = 40 nM, Kd(PRKCH) = 46 nM, Kd(FLT3(D835H)) = 49 nM, Kd(FLT3(ITD)) = 72 nM, Kd(MAPK15) = 76 nM, Kd(RSK2Kd) = 87 nM, 18 other targets with Kd 0.1-1 µM;

Screened at 1 µM, follow-up Kds for closest targets: Kd(GSK3B) = 8.3 nM, Kd(PRKCD) = 25 nM, Kd(RSK4) = 25 nM, Kd(PRKCQ) = 36 nM, Kd(FLT3(D835Y)) = 40 nM, Kd(PRKCH) = 46 nM, Kd(FLT3(D835H)) = 49 nM, Kd(FLT3(ITD)) = 72 nM, Kd(MAPK15) = 76 nM, Kd(RSK2Kd) = 87 nM, 18 other targets with Kd 0.1-1 µM;

Screened at 1 µM, follow-up Kds for closest targets: Kd(GSK3B) = 8.3 nM, Kd(PRKCD) = 25 nM, Kd(RSK4) = 25 nM, Kd(PRKCQ) = 36 nM, Kd(FLT3(D835Y)) = 40 nM, Kd(PRKCH) = 46 nM, Kd(FLT3(D835H)) = 49 nM, Kd(FLT3(ITD)) = 72 nM, Kd(MAPK15) = 76 nM, Kd(RSK2Kd) = 87 nM, 18 other targets with Kd 0.1-1 µM;

Screened at 1 µM, follow-up Kds for closest targets: Kd(GSK3B) = 8.3 nM, Kd(PRKCD) = 25 nM, Kd(RSK4) = 25 nM, Kd(PRKCQ) = 36 nM, Kd(FLT3(D835Y)) = 40 nM, Kd(PRKCH) = 46 nM, Kd(FLT3(D835H)) = 49 nM, Kd(FLT3(ITD)) = 72 nM, Kd(MAPK15) = 76 nM, Kd(RSK2Kd) = 87 nM, 18 other targets with Kd 0.1-1 µM;