ChEMBL ligand: CHEMBL2338329 (ASP-1517, ASP1517, Evrenzo, FG-4592, FG4592, Roxadustat)
Alpha-ketoglutarate-dependent dioxygenase FTO in Human [ChEMBL: CHEMBL2331065] [UniProtKB: Q9C0B1] There should be some charts here, you may need to enable JavaScript!
egl-9 family hypoxia inducible factor 1/Egl nine homolog 1 in Human [ChEMBL: CHEMBL5697] [GtoPdb: 2833] [UniProtKB: Q9GZT9] There should be some charts here, you may need to enable JavaScript!
Hypoxia-inducible factor 1-alpha inhibitor in Human [ChEMBL: CHEMBL5909] [UniProtKB: Q9NWT6] There should be some charts here, you may need to enable JavaScript!
Methylcytosine dioxygenase TET1 in Human [ChEMBL: CHEMBL4523402] [UniProtKB: Q8NFU7] There should be some charts here, you may need to enable JavaScript!
Methylcytosine dioxygenase TET2 in Human [ChEMBL: CHEMBL4523344] [UniProtKB: Q6N021] There should be some charts here, you may need to enable JavaScript!
Methylcytosine dioxygenase TET3 in Human [ChEMBL: CHEMBL4879414] [UniProtKB: O43151] There should be some charts here, you may need to enable JavaScript!
Prolyl 3-hydroxylase OGFOD1 in Human [ChEMBL: CHEMBL4523398] [UniProtKB: Q8N543] There should be some charts here, you may need to enable JavaScript!
Prolyl 4-hydroxylase in Paramecium bursaria Chlorella virus 1 [ChEMBL: CHEMBL4523368] [UniProtKB: Q84406] There should be some charts here, you may need to enable JavaScript!
Thyroid hormone receptor-α/Thyroid hormone receptor alpha in Human [ChEMBL: CHEMBL1860] [GtoPdb: 588] [UniProtKB: P10827] There should be some charts here, you may need to enable JavaScript!
Thyroid hormone receptor-β/Thyroid hormone receptor beta in Human [ChEMBL: CHEMBL1947] [GtoPdb: 589] [UniProtKB: P10828] There should be some charts here, you may need to enable JavaScript!

DB	Assay description	Assay Type	Standard value	Standard parameter	Original value	Original units	Original parameter	Reference
Alpha-ketoglutarate-dependent dioxygenase FTO in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2331065] [UniProtKB: Q9C0B1]
ChEMBL	Inhibition of human hexahistidine-tagged full-length FTO expressed in Escherichia coli BL21 (DE3) using 3-methylthymidine as substrate assessed as inhibition of 3-methylthymidine conversion to thymidine after 1 hr by liquid chromatographic analysis	B	5.01	pIC50	9800	nM	IC50	J Med Chem (2013) 56: 3680-3688 [PMID:23547775]
egl-9 family hypoxia inducible factor 1/Egl nine homolog 1 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL5697] [GtoPdb: 2833] [UniProtKB: Q9GZT9]
ChEMBL	Inhibition of HIF-PHD2 (181 to 426 residues) (unknown origin) using HIF1-alpha (556 to 574 residues) as substrate in presence of 2-OG preincubated for 30 mins followed by OPD addition measured after 10 mins by fluorescence assay	B	5.24	pIC50	5740	nM	IC50	J Med Chem (2018) 61: 5332-5349 [PMID:29856623]
ChEMBL	HIF-PH Assay: Ketoglutaric acid α-[1-14C]-sodium salt, alpha-ketoglutaric acid sodium salt, and HPLC purified peptide were obtained from commercial sources, e.g., Perkin-Elmer (Wellesley Mass.), Sigma-Aldrich, and SynPep Corp. (Dublin Calif.), respectively. Peptides for use in the assay were fragments of HIFα as described above or as disclosed in International Publication WO 2005/118836, incorporated by reference herein. For example, a HIF peptide for use in the HIF-PH assay was [methoxycoumarin]-DLDLEALAPYIPADDDFQL-amide. HIF-PH, e.g., HIF-PH2 (also known as EGLN1 or PHD2), was expressed in, e.g., insect Hi5 cells, and partially purified, e.g., through a SP ion exchange chromatography column. Enzyme activity was determined by capturing 14CO2 using an assay described by Kivirikko and Myllyla (1982, Methods Enzymol. 82:245-304). Assay reactions contained 50 mM HEPES (pH 7.4), 100 μM α-ketoglutaric acid sodium salt, 0.30 μCi/mL α-ketoglutaric acid α-[1-14C]-sodium salt, 40 μM FeSO4, 1 mM ascorbate, 1541.8 units/mL Catalase, with or without 50 μM peptide substrate and various concentrations of compound of the invention. Reactions were initiated by addition of HIF-PH enzyme.	B	5.68	pIC50	2100	nM	IC50	US-9340511-B2. Process for making isoquinoline compounds (2016)
ChEMBL	HIF-PH Assay: Ketoglutaric acid alpha -[1-14C]-sodium salt, alpha-ketoglutaric acid sodium salt, and HPLC purified peptide were obtained from commercial sources, e.g., Perkin-Elmer (Wellesley Mass.), Sigma-Aldrich, and SynPep Corp. (Dublin Calif.), respectively. Peptides for use in the assay were fragments of HIFalpha as described above or as disclosed in International Publication WO 2005/118836, incorporated by reference herein. For example, a HIF peptide for use in the HIF-PH assay was [methoxycoumarin]-DLDLEALAPYIPADDDFQL-amide. HIF-PH, e.g., HIF-PH2 (also known as EGLN1 or PHD2), was expressed in, e.g., insect Hi5 cells, and partially purified, e.g., through a SP ion exchange chromatography column. Enzyme activity was determined by capturing 14CO2 using an assay described by Kivirikko and Myllyla (1982, Methods Enzymol. 82:245-304). Assay reactions contained 50 mM HEPES (pH 7.4), 100 uM alpha -ketoglutaric acid sodium salt, 0.30 uCi/mL alpha -ketoglutaric acid alpha -[1-14C]-sodium.	B	5.68	pIC50	2100	nM	IC50	US-9115085-B2. Crystalline forms of a prolyl hydroxylase inhibitor (2015)
ChEMBL	EGLN-1 Activity Assay : The EGLN-1 (or EGLN-3) enzyme activity is determined using mass spectrometry (matrix-assisted laser desorption ionization, time-of-flight MS, MALDI-TOF MS for assay details, see reference (Greis et al., 2006). Recombinant human EGLN-1-179/426 is prepared as described above and in the Supplemental Data. Full-length recombinant human EGLN-3 is prepared in a similar way, however it is necessary to use the His-MBP-TVMV EGLN-3 fusion for the assay due to the instability of the cleaved protein. For both enzymes, the HIF-1C. peptide corresponding to residues 556-574 (DLDLEALAPYIPAD DDFQL) is used as substrate. The reaction is conducted in a total volume of 50 uL containing TrisCl (5 mM, pH 7.5), ascorbate (120 uM), 2-oxoglutarate (3.2 uM), HIF-1C. (8.6 uM), and bovine serum albumin (0.01%). The enzyme, quantity predetermined to hydroxylate 20% of substrate in 20 minutes, is added to start the reaction. Where inhibitors are used, compounds are prepared in dimethyl Sulfoxide at 10-fold final assay concentration. After 20 minutes at room temperature, the reaction is stopped by transferring 10 LIL of reaction mixture to 50 LL of a mass spectrometry matrix Solution (C-cyano-4-hydroxycinnamic acid, 5 mg/mL in 50% acetonitrile/0.1% TFA, 5 mM NHPO). Two micro liters of the mixture is spotted onto a MALDI-TOF MS target plate for analysis with an Applied Biosystems (Foster City, Calif.) 4700 Proteomics Analyzer MALDI-TOF MS equipped with a Nd:YAG laser (355 nm, 3 ns pulse width, 200 HZ repetition rate). Hydroxylated peptide product is identified from substrate by the gain of 16 Da. Data defined as percent conversion of Substrate to product is analyzed in GraphPad Prism 4 to calculate ICso values.	B	6.22	pIC50	599	nM	IC50	US-10889546-B2. Alkynyl pyridine prolyl hydroxylase inhibitor, and preparation method and medical use thereof (2021)
ChEMBL	Displacement of FITC-HIF-1alpha (556 to 574 residues) from PHD2 (181 to 426 residues) (unknown origin) after 60 mins by fluorescence polarization assay	B	6.23	pIC50	591.4	nM	IC50	ACS Med Chem Lett (2015) 6: 1236-1240 [PMID:26713111]
ChEMBL	Inhibition of HIF-PHD2 (181 to 426 residues) (unknown origin) using FITC-HIF1-alpha (556 to 574 residues) as substrate after 60 mins by fluorescence polarization assay	B	6.23	pIC50	591.4	nM	IC50	J Med Chem (2018) 61: 5332-5349 [PMID:29856623]
ChEMBL	Inhibition of FITC-HIF1alpha (556 to 574 residues) binding to PHD2 (181 to 426 residues) (unknown origin) by fluorescence polarization assay	B	6.23	pIC50	591	nM	IC50	J Med Chem (2020) 63: 10045-10060 [PMID:32787144]
ChEMBL	Inhibition of PHD2 (181 to 426 residues) (unknown origin) using FITC-HIF1alpha (556 to 574 residues) as substrate after 60 mins by fluorescence polarization assay	B	6.24	pIC50	575.3	nM	IC50	J Med Chem (2019) 62: 7583-7588 [PMID:31244107]
ChEMBL	Inhibition of PHD2 (unknown origin) measured by fluorescence polarization assay	B	6.28	pIC50	520	nM	IC50	Eur J Med Chem (2022) 230: 114115-114115 [PMID:35033824]
ChEMBL	Inhibition of PHD2 (181 to 426 residue) (unknown origin) measured by fluorescence polarization assay	B	6.92	pIC50	120.5	nM	IC50	Eur J Med Chem (2022) 238: 114479-114479 [PMID:35675755]
ChEMBL	Binding affinity to full length PHD2 (181 to 426 residues) (unknown origin) by fluorescence polarization assay	B	6.93	pIC50	117.2	nM	IC50	J Med Chem (2023) 66: 8545-8563 [PMID:37367818]
ChEMBL	Inhibition of PHD2 (unknown origin) incubated for 15 mins by competitive fluorescence polarization assay	B	7.33	pIC50	47	nM	IC50	J Med Chem (2022) 65: 7193-7211 [PMID:35507418]
ChEMBL	Inhibition of HIF-PHD2 (unknown origin)	B	7.49	pIC50	32	nM	IC50	J Med Chem (2016) 59: 11039-11049 [PMID:28002958]
ChEMBL	Inhibition of recombinant human PHD2 using 2OG as substrate and Fe2 as co-factor assessed as hydroxylation incubated for 15 mins in presence of L-ascorbate by LC-MS analysis	B	7.57	pIC50	27	nM	IC50	Bioorg Med Chem (2019) 27: 2405-2412 [PMID:30737136]
ChEMBL	Inhibition of PHD2 (unknown origin) preincubated for 30 mins followed by substrate addition and measured after 30 mins by HTS plate reader analysis	B	7.85	pIC50	14	nM	IC50	J Med Chem (2024) 67: 1393-1405 [PMID:38189253]
ChEMBL	Inhibition of PHD2 in human Hep3B cells assessed as increase in EPO production measured after 24 hrs by ELISA	B	5.21	pEC50	6110	nM	EC50	Bioorg Med Chem Lett (2022) 76: 129007-129007 [PMID:36174835]
Hypoxia-inducible factor 1-alpha inhibitor in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL5909] [UniProtKB: Q9NWT6]
ChEMBL	Inhibition of recombinant human FIH using 2OG as substrate and Fe2 as co-factor assessed as hydroxylation incubated for 15 mins in presence of L-ascorbate by LC-MS analysis	B	4	pIC50	>100000	nM	IC50	Bioorg Med Chem (2019) 27: 2405-2412 [PMID:30737136]
Methylcytosine dioxygenase TET1 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4523402] [UniProtKB: Q8NFU7]
ChEMBL	Inhibition of N-terminal 3xFlag-tagged human TET1 catalytic domain (E1418 to V2136 residues) expressed in Sf9 cells using 5-methylcytosine as substrate preincubated for 10 mins followed by DNA-cofactor addition and measured after 30 mins by Alphascreen assay	B	4	pIC50	>=100000	nM	IC50	J Med Chem (2024) 67: 4525-4540 [PMID:38294854]
ChEMBL	Inhibition of N-terminal 3xFlag-tagged human TET1 catalytic domain (E1418 to V2136 residues) expressed in Sf9 cells using 5-methylcytosine as substrate preincubated for 10 mins followed by DNA-cofactor addition and measured after 30 mins by Alphascreen assay	B	4	pIC50	>=100000	nM	IC50	J Med Chem (2024) 67: 4525-4540 [PMID:38294854]
Methylcytosine dioxygenase TET2 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4523344] [UniProtKB: Q6N021]
ChEMBL	Inhibition of His10-FLAG tagged human TET2 catalytic domain (Q969 to I2002 residues) expressed in Sf9 cells using 5-methylcytosine as substrate preincubated for 10 mins followed by DNA-cofactor addition and measured after 10 mins by Alphascreen assay	B	4	pIC50	>=100000	nM	IC50	J Med Chem (2024) 67: 4525-4540 [PMID:38294854]
ChEMBL	Inhibition of His10-FLAG tagged human TET2 catalytic domain (Q969 to I2002 residues) expressed in Sf9 cells using 5-methylcytosine as substrate preincubated for 10 mins followed by DNA-cofactor addition and measured after 10 mins by Alphascreen assay	B	4	pIC50	>=100000	nM	IC50	J Med Chem (2024) 67: 4525-4540 [PMID:38294854]
Methylcytosine dioxygenase TET3 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4879414] [UniProtKB: O43151]
ChEMBL	Inhibition of human TET3 catalytic domain (E824 to I1795 residues) expressed in mammalian cells using 5-methylcytosine as substrate preincubated for 10 mins followed by DNA-cofactor addition and measured after 10 mins by Alphascreen assay	B	4	pIC50	>=100000	nM	IC50	J Med Chem (2024) 67: 4525-4540 [PMID:38294854]
ChEMBL	Inhibition of human TET3 catalytic domain (E824 to I1795 residues) expressed in mammalian cells using 5-methylcytosine as substrate preincubated for 10 mins followed by DNA-cofactor addition and measured after 10 mins by Alphascreen assay	B	4	pIC50	>=100000	nM	IC50	J Med Chem (2024) 67: 4525-4540 [PMID:38294854]
Prolyl 3-hydroxylase OGFOD1 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4523398] [UniProtKB: Q8N543]
ChEMBL	Inhibition of recombinant human OGFOD1 using 2OG as substrate and Fe2 as co-factor assessed as hydroxylation incubated for 15 mins in presence of L-ascorbate by MALDI-TOF MS analysis	B	6	pIC50	<1000	nM	IC50	Bioorg Med Chem (2019) 27: 2405-2412 [PMID:30737136]
Prolyl 4-hydroxylase in Paramecium bursaria Chlorella virus 1 (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4523368] [UniProtKB: Q84406]
ChEMBL	Inhibition of N-terminal His6-tagged recombinant Paramecium bursaria chlorella virus 1 CPH expressed in Escherichia coli Rosetta 2 (DE3) cells pre-incubated for 5 mins before 2OG as substrate and Fe2 as co-factor addition in presence of L-ascorbate and measured after 5 mins MALDI TOF MS analysis	B	5.97	pKi	1070	nM	Ki	Bioorg Med Chem (2019) 27: 2405-2412 [PMID:30737136]
ChEMBL	Inhibition of N-terminal His6-tagged recombinant Paramecium bursaria chlorella virus 1 CPH expressed in Escherichia coli Rosetta 2 (DE3) cells pre-incubated for 5 mins before 2OG as substrate and Fe2 as co-factor addition in presence of L-ascorbate and measured after 5 mins MALDI TOF MS analysis	B	5.3	pIC50	5000	nM	IC50	Bioorg Med Chem (2019) 27: 2405-2412 [PMID:30737136]
Thyroid hormone receptor-α/Thyroid hormone receptor alpha in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL1860] [GtoPdb: 588] [UniProtKB: P10827]
ChEMBL	Agonist activity at wild type human TR alpha LBD expressed in Escherichia coli BL21 (DE3) assessed as induction of N-terminal biotinylated coactivator SRC2-3 peptide recruitment by alphascreen assay	B	7.49	pEC50	32.7	nM	EC50	J Med Chem (2022) 65: 7193-7211 [PMID:35507418]
Thyroid hormone receptor-β/Thyroid hormone receptor beta in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL1947] [GtoPdb: 589] [UniProtKB: P10828]
ChEMBL	Agonist activity at human TR beta LBD (202 to 461 residues) expressed in Escherichia coli BL21 (DE3) cells assessed as induction of N-terminal biotinylated coactivator SRC2-3 peptide recruitment by alphascreen assay	B	7.89	pEC50	12.9	nM	EC50	J Med Chem (2022) 65: 7193-7211 [PMID:35507418]

ChEMBL data shown on this page come from version 36:

Zdrazil B, Felix E, Hunter F, Manners EJ, Blackshaw J, Corbett S, de Veij M, Ioannidis H, Lopez DM, Mosquera JF, Magarinos MP, Bosc N, Arcila R, Kizilören T, Gaulton A, Bento AP, Adasme MF, Monecke P, Landrum GA, Leach AR. (2024). The ChEMBL Database in 2023: a drug discovery platform spanning multiple bioactivity data types and time periods. Nucleic Acids Res., 52(D1). DOI: 10.1093/nar/gkad1004. [EPMCID:10767899] [PMID:37933841]
Davies M, Nowotka M, Papadatos G, Dedman N, Gaulton A, Atkinson F, Bellis L, Overington JP. (2015) 'ChEMBL web services: streamlining access to drug discovery data and utilities.' Nucleic Acids Res., 43(W1). DOI: 10.1093/nar/gkv352. [EPMCID:25883136]