Chemogenomic set

fold-activation(PPARD) = 19.05, 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

Screened at 10 µM against receptors, ion channels and enzymes, closest targets as % of contr.: A1 receptor (48%), Cl- channel (GABA-gated) (23%), PDE2A1 (49%), PDE5 (2%), >10000-fold selective; In vitro activity of closest targets (TR-FRET assays): IC50(BRPF1) = 3.15 µM, IC50(BRPF3) = 5.55 µM, IC50(TAF1L, BD2) = 0.106 µM, IC50(BRD4, BD1) >15 µM, IC50(BRD4,BD2) >20 µM; Screened in DSF assay against 48 bromodomains at 10 µM, closest targets: dTm(CECR2) = 3.5 K, dTm(EP300) = 3.5 K, dTm(BRD7) = 2.3 K; In vitro follow up ITC: Kd(CECR2) = 1.39 µM; Screened against31 bromodomains in (BROMOscan DiscoverX) at 100 nM, closest targets as % contr.: CREBBP (30%), TAF1L(BD2) (0.7%); Screened against 300 kinases at 10 µM, >10000-fold selective (PMID: 28402630)

Screened at 10 µM against receptors, ion channels and enzymes, closest targets as % of contr.: A1 receptor (48%), Cl- channel (GABA-gated) (23%), PDE2A1 (49%), PDE5 (2%), >10000-fold selective; In vitro activity of closest targets (TR-FRET assays): IC50(BRPF1) = 3.15 µM, IC50(BRPF3) = 5.55 µM, IC50(TAF1L, BD2) = 0.106 µM, IC50(BRD4, BD1) >15 µM, IC50(BRD4,BD2) >20 µM; Screened in DSF assay against 48 bromodomains at 10 µM, closest targets: dTm(CECR2) = 3.5 K, dTm(EP300) = 3.5 K, dTm(BRD7) = 2.3 K; In vitro follow up ITC: Kd(CECR2) = 1.39 µM; Screened against31 bromodomains in (BROMOscan DiscoverX) at 100 nM, closest targets as % contr.: CREBBP (30%), TAF1L(BD2) (0.7%); Screened against 300 kinases at 10 µM, >10000-fold selective (PMID: 28402630)

Screeed at 10 µM in thermal shift assays, dTm(BAZ2A) = 5.2 K, dTm(BAZ2B) = 3.8 K, dTm(CECR2) = 2.0 K; Follow-up in vitro potency (ITC): Kd(CECR2) = 1.55 µM (15-fold selective); Greater 100-fold selective for BRD4 (IC50 >50 µM, BRD4 AlphaScreen assay); Clean profile in CEREP screen against 55 targets (receptors, ion chanels) at 10 µM;

Screeed at 10 µM in thermal shift assays, dTm(BAZ2A) = 5.2 K, dTm(BAZ2B) = 3.8 K, dTm(CECR2) = 2.0 K; Follow-up in vitro potency (ITC): Kd(CECR2) = 1.55 µM (15-fold selective); Greater 100-fold selective for BRD4 (IC50 >50 µM, BRD4 AlphaScreen assay); Clean profile in CEREP screen against 55 targets (receptors, ion chanels) at 10 µM;

Highly selective in methyltransferase screen at 1 µM and 10 µM; Inactive against protein arginine methyltransferases and DNA methyltransferases; Selective against a panel of 9 epigenetic readers and chromatin binders; Screened against 125 nonepigenetic targets (CEREP panel: GPCRS, ion channels, transporters) at 10 µM, closest targets >50% inhibition: Adenosin A1 receptor (98%, Ki = 21 nM), Adenosin A2A receptor (98%, Ki = 28 nM), Cl-chanel (GABA-gated, 90%, Ki = 1.8 µM), Adenosin A2 receptor (74%, 2 µM), kappa (KOP, 62%, Ki >10 µM), NK2 (61%, Ki = 4.1 µM), delta2 (DOP, 51%, Ki = 9 µM)

Highly selective in methyltransferase screen at 1 µM and 10 µM; Inactive against protein arginine methyltransferases and DNA methyltransferases; Selective against a panel of 9 epigenetic readers and chromatin binders; Screened against 125 nonepigenetic targets (CEREP panel: GPCRS, ion channels, transporters) at 10 µM, closest targets >50% inhibition: Adenosin A1 receptor (98%, Ki = 21 nM), Adenosin A2A receptor (98%, Ki = 28 nM), Cl-chanel (GABA-gated, 90%, Ki = 1.8 µM), Adenosin A2 receptor (74%, 2 µM), kappa (KOP, 62%, Ki >10 µM), NK2 (61%, Ki = 4.1 µM), delta2 (DOP, 51%, Ki = 9 µM)

>100-fold selective against other HMTs, selective against 44 GPCRS, ion channels and transporters at 1 µM, closest targets as % of contr.: H3 (93%, Ki = 45 nM), µ opioid (74%, Ki >10 µM), a2C (67%, Ki > 10 µM), a1D (59%, Ki = 4.5 µM); Selective against 50 kinases at 10 µM

>100-fold selective against other HMTs, selective against 44 GPCRS, ion channels and transporters at 1 µM, closest targets as % of contr.: H3 (93%, Ki = 45 nM), µ opioid (74%, Ki >10 µM), a2C (67%, Ki > 10 µM), a1D (59%, Ki = 4.5 µM); Selective against 50 kinases at 10 µM

Screened at 2 µM via biolayer interferometry (BLI), weak inhibition of BET bromodomains at 2.0 µM (BRD2, BRD3, BRD4, BRDT on both BD1 and BD2), clean at screen with 200 nM; DSF assay screen against 43 bromodomain family members, closest targets: dTM(BRD2, BD1) = 1.35 K, dTM(BRD3, BD1) = 1.55 K, dTM(BRD4, BD1) = 2.09 K, dTm(BRD4, BD2) = 0.58 K; In vitro follow-up (ITC): Kd(BRD4, BD1) = 5.6 µM, Kd(BRD4, BD2) = 20 µM; Screened in CEREP selectivity panel against 104 targets (nuclear receptors, ion channels, and 32 enzyme assays (including 10 kinases, 9 proteases, and 5 phosphodiesterases) at 10 µM, closest targets as % of inhibition: a1A (89%), M2 (67%);

Screened at 2 µM via biolayer interferometry (BLI), weak inhibition of BET bromodomains at 2.0 µM (BRD2, BRD3, BRD4, BRDT on both BD1 and BD2), clean at screen with 200 nM; DSF assay screen against 43 bromodomain family members, closest targets: dTM(BRD2, BD1) = 1.35 K, dTM(BRD3, BD1) = 1.55 K, dTM(BRD4, BD1) = 2.09 K, dTm(BRD4, BD2) = 0.58 K; In vitro follow-up (ITC): Kd(BRD4, BD1) = 5.6 µM, Kd(BRD4, BD2) = 20 µM; Screened in CEREP selectivity panel against 104 targets (nuclear receptors, ion channels, and 32 enzyme assays (including 10 kinases, 9 proteases, and 5 phosphodiesterases) at 10 µM, closest targets as % of inhibition: a1A (89%), M2 (67%);

Screened at 50 µM, >1000-fold selective over other methyltransferases (G9a, EZH2, EHMT1, SUV39H2, EZH1, SUV420H1, SUV420H2, SETD8, SETD2, PRMT1, PRMT3, PRMT5, PRMT8, SETDB1, MLL1, DOT1L, WHSC1, SMYD2, DNMT1); Screened against 137 ion channels, GPCRs, and enzymes at 10 µM, closest targets: Ki(A2B) = 1.3 µM, Ki(BB3) = 8.1 µM, Ki(motilin) = 7.2 µM, Ki(KOP) = 3.8 µM

Screened at 1 µM, 5 µM and 20 µM, closest target at 20 µM: METTL21a (60% of contr.); Selective against panel of 250 enzymes (kinases, GPCRs, ion channels, transporters) at 10 µM, closest targets as % of contr.: 5HT2B (69%, Ki >15 µM), BRSK1 (56%), DLK1 (60%), MSK2 (55%), PKG2 (58%), PRKX (61%);

Screened at 1 and 10 µM, closest target NSD3 (75% at 10 µM), GSK591 is highly selective

Screened in DSF assay at 10 µM, dTm(PBRM1,BD5) = 9.1 K, dTm(SMARCA2) = 8.2 K, dTM(SMARCA4) = 7.0 K, other targets dTm <1 K; Screened in CEREP panel against 102 cellular receptors and 30 enzymes, closest targets: IC50(A1A) = 0.44 µM, IC50(A1B) = 3.6 µM, IC50(kKOP) = 13 µM, IC50(5-HAT2B) = 4.0 µM;

Screened in DSF assay at 10 µM, dTm(PBRM1,BD5) = 9.1 K, dTm(SMARCA2) = 8.2 K, dTM(SMARCA4) = 7.0 K, other targets dTm <1 K; Screened in CEREP panel against 102 cellular receptors and 30 enzymes, closest targets: IC50(A1A) = 0.44 µM, IC50(A1B) = 3.6 µM, IC50(kKOP) = 13 µM, IC50(5-HAT2B) = 4.0 µM;

Screened in DSF assay at 10 µM, dTm(PBRM1,BD5) = 9.1 K, dTm(SMARCA2) = 8.2 K, dTM(SMARCA4) = 7.0 K, other targets dTm <1 K; Screened in CEREP panel against 102 cellular receptors and 30 enzymes, closest targets: IC50(A1A) = 0.44 µM, IC50(A1B) = 3.6 µM, IC50(kKOP) = 13 µM, IC50(5-HAT2B) = 4.0 µM;

Screened at 10 µM in DSF-assay, closest target: dTm(CREBBP) = 1.18 K;

Screened at 10 µM in DSF-assay, closest target: dTm(CREBBP) = 1.18 K;

Screened at 10 µM in DSF-assay, closest target: dTm(CREBBP) = 1.18 K;

Screened at 10 µM in DSF-assay, closest target: dTm(CREBBP) = 1.18 K;

Screened at 10 µM in DSF-assay, closest target: dTm(CREBBP) = 1.18 K;

Screened at 10 µM in DSF-assay, closest target: dTm(CREBBP) = 1.18 K;

Screened at 10 µM in DSF-assay, closest target: dTm(CREBBP) = 1.18 K;

Screened at 10 µM in DSF-assay, closest target: dTm(CREBBP) = 1.18 K;

DSF-assay, dTm(CECR2) = 12.52 K, closest targets, dTM 1-2 K: BRD4(BD2), BRD7, FALZ, SMARCA4, TAF1 (BD2);

No targets confirmed outside BRD family; Screened in BROMOscan panel against 46 targets, closest target: IC50(TRIM24) = 1600 nM, 500-fold selective; Clean profile in CEREP screen against 55 ligand receptors, ion channels and transporters, at 10 µM

No targets confirmed outside BRD family; Screened in BROMOscan panel against 46 targets, closest target: IC50(TRIM24) = 1600 nM, 500-fold selective; Clean profile in CEREP screen against 55 ligand receptors, ion channels and transporters, at 10 µM

No targets confirmed outside BRD family; Screened in BROMOscan panel against 46 targets, closest target: IC50(TRIM24) = 1600 nM, 500-fold selective; Clean profile in CEREP screen against 55 ligand receptors, ion channels and transporters, at 10 µM

Screened at 1 µM, in vitro follow-up of closest targets: Kd(RIPK2) = 110 nM, Kd(ADCK3) = 190 nM, Kd(NLK) = 520 nM, Kd(ACVR1) = 980 nM; Cellular selectivity: IC50(RIPK2) = 360 nM (NanoBRET assay), others not tested;

Screened at 10 µM in DSF assay, dTM (CREBBPA/EP300A) = 9.6/ 10.4 K, closest targets: dTM(BRD4) = 2.0 K, dTM(BRD3) = 1.9 K, dTm(BRD2) = 1.4 K, dTm (BRD3) = 1.2 K; Follow-up Kds via ITC: Kd(BRD4 at BD1/ BD2) = 0.854/ 4.88 µM, Ki(BRD3 at BD1/ BD2) = 1.09/ 2.19 µM, Kd(BRD2 at BD1/BD2) = 1.01/ 1.76 µM, Kd(BRD3 at BD1/BD2) = 1.09/ 2.19µM; Screened in CEREP panel against 104 receptors, ion channels and other enzymes, closest targets: Ki(alpha2A) = 570 nM, Ki(alpha2C) = 110 nM, Ki(PAF) = 540 nM;

Screened at 10 µM in DSF assay, dTM (CREBBPA/EP300A) = 9.6/ 10.4 K, closest targets: dTM(BRD4) = 2.0 K, dTM(BRD3) = 1.9 K, dTm(BRD2) = 1.4 K, dTm (BRD3) = 1.2 K; Follow-up Kds via ITC: Kd(BRD4 at BD1/ BD2) = 0.854/ 4.88 µM, Ki(BRD3 at BD1/ BD2) = 1.09/ 2.19 µM, Kd(BRD2 at BD1/BD2) = 1.01/ 1.76 µM, Kd(BRD3 at BD1/BD2) = 1.09/ 2.19µM; Screened in CEREP panel against 104 receptors, ion channels and other enzymes, closest targets: Ki(alpha2A) = 570 nM, Ki(alpha2C) = 110 nM, Ki(PAF) = 540 nM;

Selective over 15 other lysine, arginine, and DNA methyltransferases; Less than 20% inhibition on 50 kinase targets at 10 µM; Tested in PDSP screen: less than 50% inhibition on 40 7TM targets at 10 µM, greater than 50% inhibition of 4 7TM targets at 10 µM, closest targets: Ki(H3) = 300 nM, Ki(NET) = 1500nM, Ki(Sigma 1) =4700 nM, Ki(Sigma 2) = 65 nM (Radioligand binding assay);

Selective over 15 other lysine, arginine, and DNA methyltransferases; Less than 20% inhibition on 50 kinase targets at 10 µM; Tested in PDSP screen: less than 50% inhibition on 40 7TM targets at 10 µM, greater than 50% inhibition of 4 7TM targets at 10 µM, closest targets: Ki(H3) = 300 nM, Ki(NET) = 1500nM, Ki(Sigma 1) =4700 nM, Ki(Sigma 2) = 65 nM (Radioligand binding assay);

Screened at 50 µM, >1000-fold selective (IC50 >50 µM); Screened against 8 Methyl-Lysine/Arginine binder (DSF assay), dTm(WDR5) = 18.1 K, closest target: dTm(TDRD3) = 1.8 K; Screened in CEREP panel against 215 targets (kinases, GPCRs, ion channels, transporters) at 1 µM: no kinase with >50% inhibition (IC50 >1 µM)

Selectivity on methyltransferases measured by scintillation proximity assay (SPA), selective <50 µM

Selectivity on methyltransferases measured by scintillation proximity assay (SPA), selective <50 µM

Screened against 55 human recombinant receptors at 10 µM, closest targets as % inhibition: 5-HT1A (49%), 5-HAT (74%), dopamine transporter (39%); Screened against 45 other enzymes (GPCRs, kinases, transporters, ion channels, nuclear receptors), >1000-fold selective over other closely related FAD utilizing enzymes (i.e. LSD2, MAO-A, MAO-B)

Selective; <50% inhibition at 250 mM against 10 HMT targets; >50-fold selective against other members of human MBT family: IC50(L3MBTL1) = 2 µM, IC50(L3MBTL4) = 11 µM, IC50(SFMBT) >30 µM, IC50(MBTD1) = 6 µM (Alphascreen assay); Histone demethylases: < 50% inhibition at 10 mM versus 35 targets; Bromodomains: Tm shift < 0.5 °C at 10 mM versus 12 targets (DSF assay); Kinases: < 15% inhibition at 10 mM versus 49 kinases, closest target as % of contr.: FLT3 (64%); NIMH Psychoactive Drug Screening Program Selectivity Panel (Radioligand binding assay): Ki(M1) = 97 nM, Ki(M2) = 72 nM, Ki(M3) = 890 nM, Ki(M4) = 400 nM, Ki(M5) = 4.3 µM, Ki(Alpha 2C) = 860 nM

Selective over other methyltransferases (radioisotope-based filter binding assays), closest target: IC50(EZH1) = 240 nM, others <60 µM; Screened against 44 receptors and enzymes (internal screening at GSK), closest target: IC50(A1b) = 10 µM, others >10 µM, >100-fold selective

Screened against G9a, EHMT1, PRMT3/5, SETDB1, SETD7/8, MLL, EZH2, SUV39H2, SUV420H2; Clean profile in Ricerca selectivity panel at 1 µM against 29 diverse receptors

Screened in DSF assay at 10 µM: highly selective, other targets <2 K; Screened against 130 enzymes (GPCR panel and Eurofins panel) at 10 µM, closest targets: opioid receptors, Ki(MU) = 100 nM, Ki(OPRL1) = 840 nM, Ki(kappa) = 1100 nM, and 5-HAT transporter, Ki = 220 nM

Screened in DSF assay at 10 µM: highly selective, other targets <2 K; Screened against 130 enzymes (GPCR panel and Eurofins panel) at 10 µM, closest targets: opioid receptors, Ki(MU) = 100 nM, Ki(OPRL1) = 840 nM, Ki(kappa) = 1100 nM, and 5-HAT transporter, Ki = 220 nM

Screened at 1 µM, closest target as % of contr.: RIOK3 (46%); Screened at 10 µM against Eurofins-Panlabs screen against 68 targets, closest targets as % of inhibition: SLC6A2 (NET, 78%), SLC6A3 (DAT, 71 %); Screened at 1 µM, 10 µM, and 50 µM, against 32 methyltransferases (enzyme activity assays, including closely related family members SMYD3, SUV420H1, and SUV420H2) >100-fold selective; IC50(SMYD3) = 3 μM, IC50(PAR1) = 1.7 µM

Screened at 1 µM and 10 µM, highly selective for SMYD3 with no activity on other methyltransferases; Screened against 31 kinases in enzyme assays, closest target T-Fyn (HTRF assay) = 5.71 µM, others >20 µM;

Screened at 10 µM in DSF assays, dTm(BAZ2A) = 4.1 K, dTm(BAZ2B) = 2.7 K, closest targets: dTm(TAF1L) = 3.4 K, dTm(BRD9) = 2.3 K; In vitro follow up (biolayer interferometry experiment) at 0.2 µM: Kd(BAZ2B) = 0.1 µM, Kd(BRD9) = 1.2 µM, Kd(TAF1L) = 3.19 µM, (ITC experiment): Kd(TAF1L) = 3.2 µM, Kd(BRD9) = 1.1 µM; Screened in CEREP panel against 55 receptors and ion channels at 10 µM, closest target as % of contr.: Melatonin (MT-1) receptor(4%)

Screened at 10 µM in DSF assays, dTm(BAZ2A) = 4.1 K, dTm(BAZ2B) = 2.7 K, closest targets: dTm(TAF1L) = 3.4 K, dTm(BRD9) = 2.3 K; In vitro follow up (biolayer interferometry experiment) at 0.2 µM: Kd(BAZ2B) = 0.1 µM, Kd(BRD9) = 1.2 µM, Kd(TAF1L) = 3.19 µM, (ITC experiment): Kd(TAF1L) = 3.2 µM, Kd(BRD9) = 1.1 µM; Screened in CEREP panel against 55 receptors and ion channels at 10 µM, closest target as % of contr.: Melatonin (MT-1) receptor(4%)

>1600-fold selective, closest targets: pKd(BRPF2/BRD2) = 6.29, pKd(BRPF3) = 6.00, pKd(BRD9) = 6.00, pKd(BRD7) = 6.09; Screened against 48 ion channels, GPCRs and other enzymes, all pXC50 <5, compound is highly selective;

Screened in DSF assay at 10 µM, closest target: dTm(CECR2) = 2 K, follow up in Alpha Screen: IC50(CECR2) = 2.32 µM

Screened in DSF assay at 10 µM, closest target: dTm(CECR2) = 2 K, follow up in Alpha Screen: IC50(CECR2) = 2.32 µM

Screened at 1 µM and 10 µM, selective over other targets including closely related arginine methyltransferases PRMT4, PRMT6, PRMT7;

Screened at 10 µM, closest targets as % control binding: Cl-channel, GABA gated (70%), Adenosine A1 (82%); Selectivity against histone demethylases, in-vitro potency (AlphaScreen assay): IC50(KDM4A) = 100 µM, IC50(KDM4C) = 100 µM, IC50(KDM4E) >100 µM, IC50(KDM6B) >100 µM, IC50(KDM3A) >100 µM, IC50(KDM2A) = 52 µM, IC50(KDM5C) = 159 µM

In-vitro potencies of closest targets: IC50(PRMT6) = 1.3 µM, IC50(PRMT8) = 8.1 µM, IC50(PRMT4) <0.01 µM, IC50(PRMT1) >10 µM, IC50(PRMT3) >10 µM, IC50(PRMT5) >10 µM, IC50(PRMT7) >10 µM, IC50(PRMT9) >10 µM, IC50(G9a) >10 µM, IC50(GLP) >10 µM, IC50(SETDB1) >10 µM, IC50(SUV39H1) >10 µM, IC50(SUV39H2) >10 µM, IC50(PRDM9) >10 µM, IC50(SETD) >10 µM, IC50(SETD7) >10 µM, IC50(MLL1) >10 µM, IC50(MLL3) >10 µM, IC50(SETD2) >10 µM, IC50(PRC2) >10 µM, IC50(SUV420H1) >10 µM, IC50(SUV420H2) >10 µM, IC50(SMYD2) >10 µM, IC50(SMYD3) >10 µM, IC50(DNMT1) >10 µM, IC50(DNMT3A/3L) >10 µM, IC50(DNMT3B/3L) >10 µM, IC50(NSD1) >10 µM, IC50(NSD2) >10 µM, IC50(NSD3) >10 µM, IC50(ASH1L) >10 µM, IC50(DOT1L) >10 µM

Screened against 71 receptors and ion channels and against 25 other enzymes at 10 µM, closest targets as % of contr.: Adenosine A1 receptor (14%), Benzodiazepine receptor (47%), PDE2A1 (h) (25%), PDE3A (h) (48%) and PDE4D2 (h) (28%); Selectivity >30 fold over all other bromodomain family members except BRD7: Kd(BRD7) = 340 nM (ITC)

Screened at 1 µM, no significant activities below 50%

Screened at 1 µM, no significant activities below 50%

Screened in DSF assay at 10µM, dTm(PRDMT9) = 5.2K, closest target dTm(PRDMT7) = 5.0K; Screened against 108 enzymes and receptors (Eurofins Panlabs Discovery Services), closest targets>50% at 10µM: MOR, A2AB, M2, H3; In vitro follow up with GloSensor assay (PDSP screen): EC50>10µM for all targets

Screened at 1 µM, closest targets as % inhibition: ABL1(49%) and ABL2 (49%);

Screened at 10 µM, no cross reactivity among proteases owing to allosteric mechanism; Screened against 468 kinases(KinomeScan DiscoverX) at 1 µM, closest targets as % contr.: FLT3(N841I) (23%), CDK8 (39%), HIPK1 (40%), NEK3 (40%), MAST1 (43%), PIK3CA(E545K) (48%); Clean profile in screen against 45 GPCRs (PDSP screen) at 10 µM; Screened in kinase panel against 50 targets at 10 µM, closest targets: IC50(FLT3) = 3.5 µM; Screened against panel of 110 receptors, ion channels, enzymes and transporters, at 10 µM, closest target as % of inhibition: adenosine transporter AdT (52%);

Screened at 10 µM, closest targets in the GPCR scan: Ki(SIGMAR1) = 1431.53 nM, Ki(TMEM97) = 2051.16 nM, Ki(OPRK1) = 7634.84 nM; hTSH antagonism: IC50 = 24 µM; hFSH antagonism: IC50 > 16 µM; Screened against 468 kinases (KinomeScan (DiscoverX)) at 1 µM, closest target as % of contr.: CDKL2 (33%)

290-fold selectivity relative to the BD1 domains of BRD2, BRD3 and BRD4; Clean in screen against 75 kinases

290-fold selectivity relative to the BD1 domains of BRD2, BRD3 and BRD4; Clean in screen against 75 kinases

290-fold selectivity relative to the BD1 domains of BRD2, BRD3 and BRD4; Clean in screen against 75 kinases

290-fold selectivity relative to the BD1 domains of BRD2, BRD3 and BRD4; Clean in screen against 75 kinases

>70-fold selective over other bromodomains tested, closest targets: Kd(CECR2) = 140 nM, Kd(BRD7) = 390 nM, Kd(CREBBP) = 740 nM, Kd(EP300) = 770 nM, others >1.5 µM; Inactive against a panel of 49 human receptors, ion channels, kinases and other enzymes

Screened at 1 µM, closest targets as % of contr.: FER (38%), EPHB3 (42%), EPHA5 (57%); Screened against 468 kinases (KinomeScan DiscoverX) at 1 µM, closest targets as % of contr.: EPHB6 (18%), EGFR(S752-I759del) (38%), LTK (49%); Screened against 45 GPCR targets (PDSP screen) at 10 µM, closest target: Ki(SIGMAR1) = 2.50 µM

Screened at 10 µM, > 700-fold selective for CHRM2, CHRM3, CHRM4; Screened against 468 kinases (KinomeScan DiscoverX) at 1 µM, closest targets as % of contr.: PIK3CB (36%), TGFBR1 (41%), BLK (49%); Panlabs screen at 10 µM against 140 targets, closest target: IC50( PTPN2) = 10 µM;

Screened at 1 µM, closest targets: Kd(CERC2) = 66 nM, Kd(GCN5L2) = 62 nM, Kd(PCAF) = 74 nM; Follow-up IC50s in cellular NanoBRET assay: IC50(CERC2/GCN5L2/PCAF) >10 µM; DSF screen against 48 human bromodomains, dTm(BPTF) = 6.16 K, closest targets: dTm(CERC2) = 2.48 K, dTm(BRD7) = 1.95 K, dTM(PCAF) = 1.53 K; Screened in NIBR principial panel against 12 GPCRs, 3 nuclear receptors, 3 transporters and 7 other enzymes, closest tagets: IC50(Ad3) = 4.9 µM, IC50(D3) = 3.5 µM, IC50(H3) = 4 µM, others >30 µM, Screened in NIBR kinase panel against 48 kinases, no target with IC50 <30 µM;

In-vitro potencies of closest targets: IC50(PRMT6) = 1.3 µM, IC50(PRMT8) = 8.1 µM, IC50(PRMT4) <0.01 µM, IC50(PRMT1) >10 µM, IC50(PRMT3) >10 µM, IC50(PRMT5) >10 µM, IC50(PRMT7) >10 µM, IC50(PRMT9) >10 µM, IC50(G9a) >10 µM, IC50(GLP) >10 µM, IC50(SETDB1) >10 µM, IC50(SUV39H1) >10 µM, IC50(SUV39H2) >10 µM, IC50(PRDM9) >10 µM, IC50(SETD) >10 µM, IC50(SETD7) >10 µM, IC50(MLL1) >10 µM, IC50(MLL3) >10 µM, IC50(SETD2) >10 µM, IC50(PRC2) >10 µM, IC50(SUV420H1) >10 µM, IC50(SUV420H2) >10 µM, IC50(SMYD2) >10 µM, IC50(SMYD3) >10 µM, IC50(DNMT1) >10 µM, IC50(DNMT3A/3L) >10 µM, IC50(DNMT3B/3L) >10 µM, IC50(NSD1) >10 µM, IC50(NSD2) >10 µM, IC50(NSD3) >10 µM, IC50(ASH1L) >10 µM, IC50(DOT1L) >10 µM

Closest targets: Ki(BRPF3) = 100 nM, Kd(BRD1) = 110 nM, Kd(FALZ) = 130 nM, Kd(BRPF1) = 140 nM, Kd(BAZB) = 840 nM, Kd(BRD7) = 1500 nM, Kd(BRD9) = 1400 nM, others >20 µM; Screened against 50 receptors, ion channels and other enzymes (internal GSK panel), closest target: XC50(AChEase) = 3.2 µM, others >4 µM

Closest targets: Ki(BRPF3) = 100 nM, Kd(BRD1) = 110 nM, Kd(FALZ) = 130 nM, Kd(BRPF1) = 140 nM, Kd(BAZB) = 840 nM, Kd(BRD7) = 1500 nM, Kd(BRD9) = 1400 nM, others >20 µM; Screened against 50 receptors, ion channels and other enzymes (internal GSK panel), closest target: XC50(AChEase) = 3.2 µM, others >4 µM

Screened at 10 µM (binding assay), closest targets as % inhibition: PPARG (99.1%), CCKAR (98.9%), UTS2R (94.9%), OPRD1 (77.9%), MAOA (74.5%), ADORA3 (71.6%), HRH1 (69.2%), PTGS2 (62.2%); Screened against 468 kinase targets (KinomeScan DiscoverX) at 1 µM, closest targets as % contr.: AURKC (9%), MARK3 (38%); Screened against 45 GPCR targets (PDSP screen) at 10 µM, closest targets: Ki(SIGMAR1) = 136.4 nM, Ki(GABA/PBR) = 793.96 nM, Ki(TMEM97) = 806.12 nM, Ki(HTR3A) = 1142.6 nM, Ki(HRH3) = 1806.76 nM, Ki(OPRD1) = 2118.36 nM; Screened in Eurofins-Cerep panel against 7 targets at 10 µM (enzymes and uptake assays): clean profile;

Screened at 10 µM (binding assays), closest targets as % inhibition: Ca2+ channel (L, dihydropyridine site); Screened against 468 kinases (KinomeScan DiscoverX) at 1 µM, closest targets as % of contr.: HIPK4 (4.4%), MAPK12 (47%); Screened against 45 GPCR targets (PDSP screen) at 10 µM, closest target: Ki(TMEM97) = 1491.08 nM; Screened in Eurofins-Cerep panel against 7 targets at 10 µM (enzymes and uptake assays): clean profile;

Screened at 500 µM against 10 PWWP, 4 WD40, 3 Tudor and 4 MBT domains of 20 proteins (DSLS measurements): dTagg(NSD3@PWWP1) = 0.7 K, dTagg(NSD2@PWWP1) = 0.1 K, dTagg(NSD3@PWWP2) = 0.5 K, dTagg(BRPF1@PWWP) = 0.0 K, dTagg(BRD1@PWWP) = -2.2 K, dTagg(BRPF3@PWWP) = 0.3 K, dTagg(ZCWPW1@PWWP) = 0.0 K, dTagg(DNMT3A@PWWP1) = -0.1 K, dTagg(DNMT3B@PWWP1) = 0.0 K, dTagg(MSH6@PWWP) = -0.2 K, dTagg(WDR20@PWWP) = 0.0 K, dTagg(WDR5@PWWP) = - 0.7 K, dTagg(WDR48@PWWP) = 0.1 K, dTagg(EED@PWWP) = 0.2 K, dTagg(53BP1@PWWP1) = 0.1 K, dTagg(SETDB1@PWWP) = 0.9 K, dTagg(TDRD3@PWWP) = 0.8 K, dTagg(L3MBTL3@PWWP) = 0.2 K, dTagg(L3MBTL1@PWWP1) = 0.2 K, dTagg(SCML2@PWWP1) = 0.1 K (https://www.nature.com/articles/s41589-019-0310-x); Screened at 1 µM and 10 µM against 34 protein, RNA and DNA methyltransferases and EP300 (acetyltransferase): highly selective with >80% remaining activity on all proteins tested (https://www.nature.com/articles/s41589-019-0310-x); Screened at 100 µM against 48 bromodomains (DSF assay): clean selectivity profile with all targets dTm <2K (https://www.nature.com/articles/s41589-019-0310-x)

Screened at 1 µM and 10 µM, clean against a panel of 8 PRMTs, 21 protein lysine methyltransferases, 3 DNA methyltransferases, and one RNA methyltransferase, targets as % of activity in the screen at 1 µM: PRMT6 (17%), PRMT1 (>90%), PRMT3 (>90%), PRMT4 (>90%), PRMT5 (>90%), PRMT7 (>90%), PRMT8 (>90%), PRMT9 (>90%), G9a (>90%), GLP (>90%), SETDB1 (>90%), SUV39H1 (>90%), SUV39H2 (>90%), SUV420H1 (>90%), SUV420H2 (>90%), SETD7 (>90%), SETD8 (>90%), MLL1 (>90%), MLL3 (>90%), PRDM9 (>90%), PRC2 (>90%), SETD2 (>90%), SMYD2 (>90%), SMYD3 (>90%), BCDN3D (>90%), DNMT1 (>90%), DNMT3A/3L (>90%), DNMT3B/3L (>90%), DOT1L (>90%), ASH1L (>90%), NSD1 (>90%), NSD2(>90%),NSD3(>90%); Targets as % of activity in the screen at 10 µM: PRMT6 (8%), PRMT1 (>90%), PRMT3 (>90%), PRMT4 (>90%), PRMT5 (>90%), PRMT7 (>90%), PRMT8 (>90%), PRMT9 (>90%), G9a (>90%), GLP (>90%), SETDB1 (>90%), SUV39H1 (>90%), SUV39H2 (>90%), SUV420H1 (>90%), SUV420H2 (>90%), SETD7 (>90%), SETD8 (>90%), MLL1 (>90%), MLL3 (>90%), PRDM9 (>90%), PRC2 (>90%), SETD2 (>90%), SMYD2 (>90%), SMYD3 (>90%), BCDN3D (>90%), DNMT1 (>90%), DNMT3A/3L (>90%), DNMT3B/3L (>90%), DOT1L (>90%), ASH1L (>90%), NSD1 (>90%), NSD2(>90%),NSD3(>90%); Screened against 44 non-epigenetic targets by Eurofins (kinases, GPCRs, ion channels and transporters) at 1 μM, closest target as % of inhibition: KOP (20%), COX1 (17%), GR (15%), 5-HT2A (12%), M3 (12%), MAOA (12%), D1 (9%), acetylcholinesterase (7%), https://pubmed.ncbi.nlm.nih.gov/33591753/;

Screened at 1 µM and 10 µM, clean against a panel of 8 PRMTs, 21 protein lysine methyltransferases, 3 DNA methyltransferases, and one RNA methyltransferase, targets as % of activity in the screen at 1 µM: PRMT6 (17%), PRMT1 (>90%), PRMT3 (>90%), PRMT4 (>90%), PRMT5 (>90%), PRMT7 (>90%), PRMT8 (>90%), PRMT9 (>90%), G9a (>90%), GLP (>90%), SETDB1 (>90%), SUV39H1 (>90%), SUV39H2 (>90%), SUV420H1 (>90%), SUV420H2 (>90%), SETD7 (>90%), SETD8 (>90%), MLL1 (>90%), MLL3 (>90%), PRDM9 (>90%), PRC2 (>90%), SETD2 (>90%), SMYD2 (>90%), SMYD3 (>90%), BCDN3D (>90%), DNMT1 (>90%), DNMT3A/3L (>90%), DNMT3B/3L (>90%), DOT1L (>90%), ASH1L (>90%), NSD1 (>90%), NSD2(>90%),NSD3(>90%); Targets as % of activity in the screen at 10 µM: PRMT6 (8%), PRMT1 (>90%), PRMT3 (>90%), PRMT4 (>90%), PRMT5 (>90%), PRMT7 (>90%), PRMT8 (>90%), PRMT9 (>90%), G9a (>90%), GLP (>90%), SETDB1 (>90%), SUV39H1 (>90%), SUV39H2 (>90%), SUV420H1 (>90%), SUV420H2 (>90%), SETD7 (>90%), SETD8 (>90%), MLL1 (>90%), MLL3 (>90%), PRDM9 (>90%), PRC2 (>90%), SETD2 (>90%), SMYD2 (>90%), SMYD3 (>90%), BCDN3D (>90%), DNMT1 (>90%), DNMT3A/3L (>90%), DNMT3B/3L (>90%), DOT1L (>90%), ASH1L (>90%), NSD1 (>90%), NSD2(>90%),NSD3(>90%); Screened against 44 non-epigenetic targets by Eurofins (kinases, GPCRs, ion channels and transporters) at 1 μM, closest target as % of inhibition: KOP (20%), COX1 (17%), GR (15%), 5-HT2A (12%), M3 (12%), MAOA (12%), D1 (9%), acetylcholinesterase (7%), https://pubmed.ncbi.nlm.nih.gov/33591753/;

Screened at 10 µM, >10,000-fold selectivity over other P2RY* receptors, closest targets: Ki(DRD3) = 884.3 nM, Ki(TMEM97) = 1120.99 nM, Ki(SIGMAR1) = 1180.59 nM, Ki(ADRA2B) = 2561.53 nM; Screened against 468 kinases (KinomeScan DiscoverX) at 1 µM, closest target: MAPKAPK2 (51% of contr.); Screened in Eurofins-Panlabs enzyme assay against 40 targets at 10 µM, closest targets: IC50(INSR) = 4.31 µM, IC50(EGFR) = 4.59 µM; Screened in Eurofins-Panlabs radioligand binding assay against 125 targets at 1 µM and 10 µM, closest targets: IC50(CYSLTR1) = 560 nM, IC50(HRH2) = 1390 nM, IC50(SLC6A3) = 2310 nM; Screened in PDSP screen against 45 receptors, ion channels and transporters at 10 µM, closest targets: Ki(D3 dopamine receptor) = 6.79 µM, Ki(d-opioid receptor) = 2.75 µM

No related human family members

Screened at 10 µM against a panel of ion channels, GPCRs, hydrolases and other enzymes, closest target as % of contr.: NA+/SITE2/R (48%, voltage-gated ion channel) 1000-fold selective over closest family members: human SLC13A2 and SLC13A3 that share physiological substrates citrate and succinate

Selective across all bromodomains (IC50(CERC2) >40 µM, IC50(YESTS2/4) >20 µM (TR-FRET assay); Screened against 59 kinases (PSP kinase safety panel, Eurofins), closest targets: IC50(TRB2) = 3.5 µM, IC50(PIM1) = 8.6 µM, others >30 µM, Screened against 31 other targets (GPCRs, transporter and ion channels, PSP Principial panel 20118+, Eurofins), closest targets: IC50(ACES) = 250 nM, IC50(H3) = 290 nM, IC50(M2) = 1.8 µM, IC50(NET) = 8.7 µM, others >30 µM;

Selective across all bromodomains (IC50(CERC2) >40 µM, IC50(YESTS2/4) >20 µM (TR-FRET assay); Screened against 59 kinases (PSP kinase safety panel, Eurofins), closest targets: IC50(TRB2) = 3.5 µM, IC50(PIM1) = 8.6 µM, others >30 µM, Screened against 31 other targets (GPCRs, transporter and ion channels, PSP Principial panel 20118+, Eurofins), closest targets: IC50(ACES) = 250 nM, IC50(H3) = 290 nM, IC50(M2) = 1.8 µM, IC50(NET) = 8.7 µM, others >30 µM;