ChEMBL ligand: CHEMBL566315 (6-ecdca, 6-ethylchenodeoxycholic acid, Acide obeticholique, Acido obeticolico, DSP-1747, INT-747, Obeticholic acid, Ocaliva)
Farnesoid X receptor/Bile acid receptor in Human [ChEMBL: CHEMBL2047] [GtoPdb: 603] [UniProtKB: Q96RI1] There should be some charts here, you may need to enable JavaScript!
GPBA receptor/G-protein coupled bile acid receptor 1 in Human [ChEMBL: CHEMBL5409] [GtoPdb: 37] [UniProtKB: Q8TDU6] 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
Farnesoid X receptor/Bile acid receptor in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2047] [GtoPdb: 603] [UniProtKB: Q96RI1]
ChEMBL	Agonist activity at FXR (unknown origin) by coactivator recruitment assay	B	4	pEC50	99000	nM	EC50	J Med Chem (2014) 57: 8035-8055 [PMID:25255039]
ChEMBL	Agonist activity at FXR transfected in human HEK293T cells o-expressing pGL3/hBSEP/luc incubated for 24 hrs by Steady-Glo reagent based luciferase reporter gene assay	B	4.38	pEC50	42000	nM	EC50	J Med Chem (2020) 63: 3868-3880 [PMID:31940200]
ChEMBL	Agonist activity at GST-tagged human FXR-LBD (193 to 472 residues) using biotinylated SRC-1 peptide as substrate incubated for 1 hrs by HTRF assay	B	5	pEC50	10000	nM	EC50	J Med Chem (2020) 63: 3868-3880 [PMID:31940200]
ChEMBL	Agonist activity at human FXR expressed in HEK293 cells measured after 18 hrs by steady-glo luciferase reporter gene assay	B	6.15	pEC50	710	nM	EC50	J Med Chem (2020) 63: 12748-12772 [PMID:32991173]
ChEMBL	Agonist activity at Gal4-fused FXR LBD (unknown origin) transfected in HEK293T cells co-transfected with Peak12-Gal4UAS-luciferase reporter assessed as receptor transactivation incubated for 24 hrs by firefly luciferase assay	B	6.17	pEC50	670	nM	EC50	Eur J Med Chem (2023) 245: 114903-114903 [PMID:36375336]
ChEMBL	Agonist activity at Gal4-fused FXR LBD (unknown origin) transfected in HEK293T cells co-transfected with Peak12-Gal4UAS-luci assessed as receptor transactivation incubated for 24 hrs by luciferase reporter gene assay	B	6.17	pEC50	670	nM	EC50	Eur J Med Chem (2023) 252: 115307-115307 [PMID:37003047]
ChEMBL	Human FXR (NR1H4) Assay: Determination of a ligand mediated Gal4 promoter driven transactivation to quantify ligand binding mediated activation of FXR. FXR Reporter Assay kit purchased from Indigo Bioscience (Catalogue number: IB00601) to determine the potency and efficacy of compound developed by Enanta that can induce FXR activation. The principle application of this reporter assay system is to quantify functional activity of human FXR. The assay utilizes non-human mammalian cells, CHO (Chinese hamster ovary) cells engineered to express human NR1H4 protein (referred to as FXR). Reporter cells also incorporate the cDNA encoding beetle luciferase which catalyzes the substrates and yields photon emission. Luminescence intensity of the reaction is quantified using a plate-reading luminometer, Envision. Reporter Cells include the luciferase reporter gene functionally linked to an FXR responsive promoter. Thus, quantifying changes in luciferase expression in the treated reporter cells provides a sensitive surrogate measure of the changes in FXR activity. EC50 and efficacy (normalize to CDCA set as 100%) is determined by XLFit. The assay is according to the manufacturer's instructions. In brief, the assay was performed in white, 96 well plates using final volume of 200 ul containing cells with different doses of compounds. Retrieve Reporter Cells from −80° C. storage. Perform a rapid thaw of the frozen cells by transferring a 10 ml volume of 37° C. cell recovery medium into the tube of frozen cells. Recap the tube of Reporter Cells and immediately place it in a 37° C. water bath for 5-10 minutes. Retrieve the tube of Reporter Cell Suspension from the water bath. Sanitize the outside surface of the tube with a 70% alcohol swab, and then transfer it into the cell culture hood. Dispense 100 μl of cell suspension into each well of the 96-well Assay Plate. Transfer the plate into 37° C. incubator, allowing the cells adherent to the bottom of the well. Dilute compounds in Dilution Plate (DP), and administrate to cells at Assay Plate (AP). DMSO content of the samples was kept at 0.2%. Cells were incubated for additional 22 hours before luciferase activities were measured. Thirty minutes before intending to quantify FXR activity, remove Detection Substrate and Detection Buffer from the refrigerator and place them in a low-light area so that they may equilibrate to room temperature. Remove the plate's lid and discard all media contents by ejecting it into an appropriate waste container. Gently tap the inverted plate onto a clean absorbent paper towel to remove residual droplets. Cells will remain tightly adhered to well bottoms. Add 100 μl of luciferase detection reagent to each well of the assay plate. Allow the assay plate to rest at room temperature for at least 5 minutes following the addition of LDR. Set the instrument (Envision) to perform a single 5 second “plate shake” prior to reading the first assay well. Read time may be 0.5 second (500 mSec) per well. EC50 and Efficacy (normalize to CDCA set as 100%) is determined by XLFit.	B	6.26	pEC50	550	nM	EC50	US-10266560-B2. Bile acid analogs as FXR/TGR5 agonists and methods of use thereof (2019)
ChEMBL	Human FXR (NR1H4) Assay: The assay utilizes non-human mammalian cells, CHO (Chinese hamster ovary) cells engineered to express human NR1H4 protein (referred to as FXR). Reporter cells also incorporate the cDNA encoding beetle luciferase which catalyzes the substrates and yields photon emission. Luminescence intensity of the reaction is quantified using a plate-reading luminometer, Envision. Reporter Cells include the luciferase reporter gene functionally linked to an FXR responsive promoter. Thus, quantifying changes in luciferase expression in the treated reporter cells provides a sensitive surrogate measure of the changes in FXR activity. EC50 and efficacy (normalize to CDCA set as 100%) is determined by XLFit. The assay is according to the manufacturer's instructions. In brief, the assay was performed in white, 96 well plates using final volume of 100 ul containing cells with different doses of compounds. Retrieve Reporter Cells from −80° C. storage. Perform a rapid thaw of the frozen cells by transferring a 10 ml volume of 37° C. cell recovery medium into the tube of frozen cells. Recap the tube of Reporter Cells and immediately place it in a 37° C. water bath for 5-10 minutes. Retrieve the tube of Reporter Cell Suspension from the water bath. Sanitize the outside surface of the tube with a 70% alcohol swab, and then transfer it into the cell culture hood. Dispense 90 μl of cell suspension into each well of the 96-well Assay Plate. Transfer the plate into 37° C. incubator, allowing the cells adherent to the bottom of the well. Dilute compounds in Dilution Plate (DP), and administrate to cells at Assay Plate (AP). DMSO content of the samples was kept at 0.2%. Cells were incubated for additional 22 hours before luciferase activities were measured. Thirty minutes before intending to quantify FXR activity, remove Detection Substrate and Detection Buffer from the refrigerator and place them in a low-light area so that they may equilibrate to room temperature. Remove the plate's lid and discard all media contents by ejecting it into an appropriate waste container. Gently tap the inverted plate onto a clean absorbent paper towel to remove residual droplets. Cells will remain tightly adhered to well bottoms. Add 100 μl of luciferase detection reagent to each well of the assay plate. Allow the assay plate to rest at room temperature for at least 5 minutes following the addition of LDR. Set the instrument (Envision) to perform a single 5 second plate shake prior to reading the first assay well. Read time may be 0.5 second (500 mSec) per well. EC50 and Efficacy (normalize to CDCA set as 100%) is determined by XLFit.To assess the FXR agonistic potency of the exam	B	6.26	pEC50	550	nM	EC50	US-10144729-B2. Isoxazole analogs as FXR agonists and methods of use thereof (2018)
ChEMBL	Human FXR (NR1H4) Assay: The potency and efficacy of the compounds of the invention on TGR5 receptor was evaluated using in vitro assays which carried out using the express kit from DiscoverX (cAMP HUNTER™ eXpress GPBAR1 CHO-K1 GPCR Assay; Cataloguer number: 95-0049E2CP2S)GPBAR1 (G protein-coupled bile acid receptor 1) encodes a member of the G protein-coupled receptor (GPCR) superfamily. GPBAR1 activation following ligand binding initiates a series of second messenger cascades that result in a cellular response. Treatment of CHO cells expressing GPBAR1 with bile acids induces the production of intracellular cAMP and internalization of the receptor. The potency and efficacy of compound for GPBAR1 activation by measuring cyclic adenosine monophosphate (cyclic AMP or cAMP) levels in live cells using a competitive immunoassay based on Enzyme Fragment Complementation (EFC).In briefly, following seeding the cells into the white, 96 well microplate, place it in a 37° C., 5% CO2 in a humidified incubator for 18-24 hours prior to testing. On second day, proceed to the appropriate cAMP HUNTER™ eXpress Protocol according to the manufacturer's instructions. Dissolve agonist compound in DMSO at the desired stock concentration, and prepare 3-fold serial dilutions of agonist compound in Cell Assay Buffer. The concentration of each dilution should be prepared at 4× of the final screening concentration (i.e. 15 μL compound+45 μL Cell Assay Buffer/cAMP Antibody Reagent). For each dilution, the final concentration of solvent should remain constant. Transfer 15 μL diluted compound the assay plate and incubate the plate for 30 minutes at 37° C. Following agonist incubation, add 60 μL of working cAMP detection reagents/cAMP Solution mixture (cAMP Lysis Buffer, Substrate Reagent 1, cAMP Solution D) to the appropriate wells. Incubate for 1 hour at room temperature (23° C.), protected from light. Add 60 μl of cAMP Solution A to the appropriate wells. Incubate for 3 hours at room temperature (23° C.), protected from light. Read samples on Envision standard luminescence plate reader. Calculate of average EC50 after logarithm transformation.	B	6.26	pEC50	550	nM	EC50	US-10208081-B2. Bile acid derivatives as FXR/TGR5 agonists and methods of use thereof (2019)
ChEMBL	Transactivation of human FXR (unknown origin) expressed in HepG2 cells co-expressing pSG5RXR/pGL4.70 after 24 hrs post transfection by luciferase reporter gene assay	B	6.3	pEC50	500	nM	EC50	ACS Med Chem Lett (2019) 10: 407-412 [PMID:30996771]
ChEMBL	Agonist activity at human FXR expressed in COS1 cells after 5 hrs by CRE-driven luciferase reporter gene assay	F	6.44	pEC50	361	nM	EC50	J Med Chem (2007) 50: 4265-4268 [PMID:17685603]
ChEMBL	Agonist activity at human FXR	B	6.44	pEC50	360	nM	EC50	Eur J Med Chem (2023) 250: 115143-115143 [PMID:36841086]
ChEMBL	Agonist activity at human FXR expressed in human Huh-7 cells co-transfected with FXRE-Luc reporter plasmid assessed as receptor transactivation incubated for 16 hrs by luciferase reporter gene assay	B	6.49	pEC50	324.1	nM	EC50	Bioorg Med Chem (2021) 43: 116280-116280 [PMID:34256254]
ChEMBL	Affinity On-target Cellular interaction: (Reporter gene assay (HEK293T cells)) EUB0000081c NR1H4	F	6.52	pEC50	300	nM	EC50	Affinity On-target Cellular Literature for EUbOPEN Chemogenomic Library
ChEMBL	Agonist activity at GST-tagged human FXR LBD assessed as induction of biotinylated coactivator SRC-1 peptide recruitment measured after 2 hrs by streptavidin-conjugated AlphaScreen assay	B	6.54	pEC50	290	nM	EC50	J Med Chem (2020) 63: 12748-12772 [PMID:32991173]
ChEMBL	Agonist activity at human GST-fused FXR LBD assessed as coactivator interaction with receptor ligand binding domain by Alphascreen assay	B	6.7	pEC50	200	nM	EC50	Bioorg Med Chem (2011) 19: 2650-2658 [PMID:21459580]
ChEMBL	Agonist activity at recombinant human GST-tagged FXR LBD (193 to 472 residues) expressed in baculovirus-infected insect cells assessed as recruitment of biotinylated SRC1 peptide measured after 1 hr by Alphascreen assay	B	6.74	pEC50	180	nM	EC50	ACS Med Chem Lett (2017) 8: 1246-1251 [PMID:29259742]
ChEMBL	Agonist activity at human full length FXR expressed in HeLa cells cotransfected with pSG5-human RXR after 24 hrs by Dual-Glo luciferase reporter gene assay	B	6.8	pEC50	160	nM	EC50	Bioorg Med Chem (2015) 23: 3490-3498 [PMID:25934227]
ChEMBL	Agonist activity at human FXR expressed in human HeLa cells assessed as receptor activation by BSEP promoter-driven firefly luciferase reporter gene assay	B	6.8	pEC50	160	nM	EC50	J Med Chem (2014) 57: 8035-8055 [PMID:25255039]
ChEMBL	Agonist activity at glutathione transferase-tagged human FXR ligand binding domain assessed as biotinylated Src1 peptide recruitment incubated for 30 mins by AlphaScreen assay	B	6.82	pEC50	150	nM	EC50	Eur J Med Chem (2022) 242: 114652-114652 [PMID:36049273]
ChEMBL	Agonist activity at glutathione transferase-tagged human FXR-LBD using biotinylated Src-1 peptide incubated for 30 mins by recruitment coactivator assay	B	6.82	pEC50	150	nM	EC50	J Med Chem (2016) 59: 9201-9214 [PMID:27652492]
GtoPdb	-	-	7	pEC50	99	nM	EC50	J Med Chem (2002) 45: 3569-72 [PMID:12166927]
ChEMBL	Agonist activity at FXR (unknown origin) assessed as recruitment of SRC1 peptide to FXR by FRET assay	B	7	pEC50	100	nM	EC50	J Med Chem (2017) 60: 5235-5266 [PMID:28252961]
ChEMBL	Binding affinity for Farnesoid X Receptor (FXR)	B	7	pEC50	99	nM	EC50	Bioorg Med Chem Lett (2003) 13: 1865-1868 [PMID:12749886]
ChEMBL	Effective concentration against Farnesoid X receptor (FXR)	B	7	pEC50	99	nM	EC50	J Med Chem (2002) 45: 3569-3572 [PMID:12166927]
ChEMBL	Activation of human farnesoid X receptor	B	7	pEC50	99	nM	EC50	J Med Chem (2005) 48: 5383-5403 [PMID:16107136]
ChEMBL	Agonist activity at FXR expressed in COS1 cells by cell-based bioluminescence assay	F	7	pEC50	99	nM	EC50	J Med Chem (2009) 52: 7958-7961 [PMID:20014870]
ChEMBL	Agonist activity at FXR (unknown origin)	B	7	pEC50	99	nM	EC50	J Med Chem (2020) 63: 5031-5073 [PMID:31930920]
ChEMBL	Agonist activity at FXR (unknown origin) by fluorescence resonance energy tranfer assay	B	7	pEC50	99	nM	EC50	Eur J Med Chem (2020) 197: 112311-112311 [PMID:32339855]
ChEMBL	Agonist activity at FXR (unknown origin) by FRET assay	B	7	pEC50	99	nM	EC50	Eur J Med Chem (2022) 243: 114742-114742 [PMID:36155354]
ChEMBL	Binding affinity to FXR assessed as ligand-dependent SRC1 recruitment by FRET based co-activator assay	F	7.01	pEC50	98	nM	EC50	J Med Chem (2006) 49: 4208-4215 [PMID:16821780]
ChEMBL	Binding affinity for human Farnesoid X receptor in FRET assay	B	7.05	pEC50	90	nM	EC50	J Med Chem (2004) 47: 4559-4569 [PMID:15317466]
ChEMBL	Agonist activity at FXR (unknown origin) by reporter gene assay	B	7.07	pEC50	85	nM	EC50	J Med Chem (2014) 57: 8035-8055 [PMID:25255039]
ChEMBL	Agonist activity at full length FXR in human Huh-7 cells	B	7.07	pEC50	85	nM	EC50	Eur J Med Chem (2020) 197: 112311-112311 [PMID:32339855]
ChEMBL	Agonist activity at FXR (unknown origin) by cell based assay	B	7.07	pEC50	85	nM	EC50	J Med Chem (2020) 63: 5031-5073 [PMID:31930920]
ChEMBL	Coactivation of TR-FRET Farnesoid X Receptor Protein: Purchasing Invitrogen PV4833 Kit.Step 1, the compound was weighed and dissolved in 100% DMSO, the highest concentration is 3000 μM, and then diluted with DMSO by 3-fold serial dilution to get 10 concentrations.Step 2, the above formulated compound solutions were diluted 100 times with the buffer provided in the kit, which were mixed respectively and 10 μL of which were added into a 384 well plate in turn.Step 3, FXR recombinant nucleo receptor protein was diluted with a buffer to formulate 4× concentration, and 5 μL of which was added into the above 384 well plate containing the compound.Step 4, Fluorescein-SRC2-2 and Tb anti-GST antibody were diluted with a buffer respectively, both concentration are 4×, after the two reagents were mixed together, 10 μL of which was added into the 384 well plate described in step 3.At last, the solution in the 384 well plate was blend by centrifugation, and then incubated at room temperature for 1 hour. And then the detection was performed at 520, 495 and 337 nm wavelengths by TR-FRET endpoint method, ECso values were calculated using the detection values of ER=520 nm/495 nm wavelength.	B	7.12	pEC50	75	nM	EC50	US-11208418-B2. Nitrogenous tricyclic compounds and uses thereof in medicine (2021)
ChEMBL	Agonist activity at human FXR expressed in HEK293T cells assessed as BSEP promoter driven cellular transcriptional activity after 24 hrs by luciferase reporter gene assay	B	7.38	pEC50	42	nM	EC50	J Med Chem (2017) 60: 9960-9973 [PMID:29148806]
ChEMBL	Agonist activity at recombinant human GST-tagged FXR ligand binding domain (193 to 472 residues) expressed in baculovirus infected insect cells assessed as induction of interaction with biotin labelled SRC-1 after 1 hr by HTRF assay	B	8	pEC50	10	nM	EC50	J Med Chem (2017) 60: 9960-9973 [PMID:29148806]
GPBA receptor/G-protein coupled bile acid receptor 1 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL5409] [GtoPdb: 37] [UniProtKB: Q8TDU6]
ChEMBL	Agonist activity at human TGR5 receptor expressed in NCI-H716 cells assessed as intracellular cAMP level by TR-FRET assay	F	4.7	pEC50	20000	nM	EC50	Bioorg Med Chem (2011) 19: 2650-2658 [PMID:21459580]
ChEMBL	Agonist activity at TGR5 in human NCI-H716 cells assessed as stimulation of intracellular cAMP accumulation incubated for 60 mins by HTR-FRET assay	F	4.82	pEC50	15000	nM	EC50	J Med Chem (2016) 59: 9201-9214 [PMID:27652492]
ChEMBL	Agonist activity at TGR5 receptor in human NCI-H716 cells assessed as activation of intracellular cAMP production incubated for 1 hr by HTR-FRET assay	F	4.85	pEC50	14000	nM	EC50	Eur J Med Chem (2022) 242: 114652-114652 [PMID:36049273]
ChEMBL	Agonist activity at human TGR5 expressed in HEK293 cells assessed as increase in intracellular cAMP level after 1 hr by TR-FRET assay	F	6.08	pEC50	840	nM	EC50	ACS Med Chem Lett (2017) 8: 1246-1251 [PMID:29259742]
ChEMBL	Agonist activity at TGR5 (unknown origin)	F	6.12	pEC50	760	nM	EC50	Eur J Med Chem (2023) 250: 115143-115143 [PMID:36841086]
ChEMBL	Agonist activity at human TGR5 expressed in CHO cells after 5 hrs by CRE-driven luciferase reporter gene assay	F	6.12	pEC50	755	nM	EC50	J Med Chem (2007) 50: 4265-4268 [PMID:17685603]
ChEMBL	Agonist activity at wild type TGR5 (unknown origin)	B	7	pEC50	100	nM	EC50	ACS Med Chem Lett (2013) 4: 1158-1162 [PMID:24900622]

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]