ChEMBL ligand: CHEMBL67279 (PT-100, Talabostat, Valinyl-l-boroproline)
Dipeptidyl-peptidase 7/Dipeptidyl peptidase 2 in Human [ChEMBL: CHEMBL3976] [GtoPdb: 1605] [UniProtKB: Q9UHL4] There should be some charts here, you may need to enable JavaScript!
dipeptidyl peptidase 4/Dipeptidyl peptidase 4 in Human [ChEMBL: CHEMBL284] [GtoPdb: 1612] [UniProtKB: P27487] dipeptidyl peptidase 4/Dipeptidyl peptidase 4 in Mouse [ChEMBL: CHEMBL3883] [GtoPdb: 1612] [UniProtKB: P28843] There should be some charts here, you may need to enable JavaScript!
dipeptidyl peptidase 8/Dipeptidyl peptidase 8 in Human [ChEMBL: CHEMBL4657] [GtoPdb: 2356] [UniProtKB: Q6V1X1] There should be some charts here, you may need to enable JavaScript!
dipeptidyl peptidase 9/Dipeptidyl peptidase 9 in Human [ChEMBL: CHEMBL4793] [GtoPdb: 2357] [UniProtKB: Q86TI2] There should be some charts here, you may need to enable JavaScript!
Presequence protease, mitochondrial in Human [ChEMBL: CHEMBL3124731] [UniProtKB: Q5JRX3] There should be some charts here, you may need to enable JavaScript!
prolyl endopeptidase/Prolyl endopeptidase in Human [ChEMBL: CHEMBL3202] [GtoPdb: 2395] [UniProtKB: P48147] There should be some charts here, you may need to enable JavaScript!
fibroblast activation protein alpha/Prolyl endopeptidase FAP in Human [ChEMBL: CHEMBL4683] [GtoPdb: 2365] [UniProtKB: Q12884] fibroblast activation protein alpha/Prolyl endopeptidase FAP in Mouse [ChEMBL: CHEMBL5769] [GtoPdb: 2365] [UniProtKB: P97321] There should be some charts here, you may need to enable JavaScript!
peptidase D/Xaa-Pro dipeptidase in Human [ChEMBL: CHEMBL4185] [GtoPdb: 2389] [UniProtKB: P12955] 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
Dipeptidyl-peptidase 7/Dipeptidyl peptidase 2 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL3976] [GtoPdb: 1605] [UniProtKB: Q9UHL4]
ChEMBL	Inhibitory activity of compound against Dipeptidylpeptidase II (DPP II)	B	3.9	pKi	125000	nM	Ki	J Med Chem (2003) 46: 5005-5014 [PMID:14584950]
ChEMBL	Inhibition of Dipeptidyl Peptidase II (Quiescent cell proline peptidase)	B	6.9	pKi	125	nM	Ki	Bioorg Med Chem Lett (2002) 12: 2825-2828 [PMID:12270155]
ChEMBL	Inhibition of quiescent cell proline dipeptidase (QPP)	B	6.9	pKi	125	nM	Ki	J Med Chem (2004) 47: 4135-4141 [PMID:15293982]
ChEMBL	In Vitro Inhibition Assay: EnzymesRecombinant human DPPIV (R&D Systems, Cat. No. 1180-SE)Recombinant human DPP8 (Enzo Life Sciences, Cat. No. BML-SE527)Recombinant human DPP9 (R&D Systems, Cat. No. 5419-SE)Recombinant human DPPII (R&D Systems, Cat. No. 3438-SE)Recombinant human FAP (R&D Systems, Cat. No. 3715-SE)Recombinant human PREP (R&D Systems, Cat. No. 4308-SE)Assay Buffers25 mM Tris, pH 8.0 (DPPIV and DPP9)50 mM Tris, pH 7.5 (DPP8)25 mM MES, pH 6.0 (DPPII)50 mM Tris, 140 mM NaCl, pH 7.5 (FAP)25 mM Tris, 0.25 M NaCl, pH 7.5 (PREP)Substrates4000× substrate solution (100 mM Gly-Pro-AMC (VWR, Cat. No. 100042-646) in DMSO, DPPIV, DPP8 and DPP9)4000× substrate solution (100 mM Lys-Pro-AMC (Bachem, Cat. No. I-1745) in DMSO, DPPII)100× substrate solution (2.5 mM Z-Gly-Pro-AMC (VWR, Cat. No. I-1145.0050BA) in DMSO, FAP and PREP)General MaterialsCompound96-well black clear-bottom plates (Costar, Cat. No. 3603)InstrumentationPlate shakerMolecular Devices SpectraMax® M2e microplate readerProtocol1. To prepare the compound for the assay, dissolve it in either DMSO or, if cyclization is suspected, in pH 2.0 water (0.01 N HCl) to a final concentration of 100 mM. For pH 2.0 stocks, incubate at room temperature for a minimum of four hours and up to overnight. From this, prepare a 1 mM stock at pH 7.4 in 50 mM Tris. If the inhibitor is insoluble at this concentration, dilute the 100 mM stock 1:10 to 10 mM. Using this stock, prepare a 0.1 mM stock as described above.2. Prepare a dilution plate for the compound stocks to be tested. Add the 0.1 and/or 1 mM stocks prepared previously to row A of a 96-well plate. From this, perform 1:10 serial dilutions into the appropriate assay buffer down the columns as shown below:3. Prepare 20× substrate solution by diluting the DMSO stocks into the appropriate assay buffer.4. Dilute the enzymes into their appropriate assay buffers. The dilution factor is lot dependent and must be determined prior to performing the assay. The final enzyme concentrations should be 0.1, 0.8, 0.4, 0.2, 1.2, and 0.6 nM for DPPIV, 8, 9, II, FAP and PREP respectively. Add 180 μL to each well needed in columns 2-10.5. Add 20 μL of the compound of interest from the dilution plate prepared in step 2 to columns 2-10 of the assay plate where appropriate. Each sample should be tested in triplicate. Allow this to incubate for 10 minutes at room temperature, shaking the plate for the first two minutes.6. Add 10 μL of 20× substrate prepared in step 3 to each well and allow this to incubate for 15 minutes at room temperature, shaking the plate for the first two minutes.	B	5.09	pIC50	8200	nM	IC50	US-11096924-B2. Combination therapies using immuno-dash inhibitors and PGE2 antagonists (2021)
ChEMBL	Inhibition of human seminal plasma DPP2 assessed as pNA release from Lys-Ala-p-nitroanilide substrate pre-incubated with enzyme for 15 min prior to substrate addition by fluorescence technique	B	7.07	pIC50	86	nM	IC50	Bioorg Med Chem Lett (2012) 22: 3412-3417 [PMID:22525314]
ChEMBL	Inhibition of DPP2 in human seminal plasma using Lys-Ala-p-nitroanilide as substrate incubated for 15 mins prior to substrate addition	B	7.07	pIC50	86	nM	IC50	ACS Med Chem Lett (2013) 4: 491-496 [PMID:24900696]
ChEMBL	Inhibition of DPP2 (unknown origin) using AMC substrate by fluorometric assay	B	7.07	pIC50	86	nM	IC50	Bioorg Med Chem Lett (2021) 37: 127846-127846 [PMID:33571650]
ChEMBL	Inhibition of DPP2 purified from human seminal plasma using Lys-Ala-p-nitroanilide as substrate by spectrophotometry	B	7.07	pIC50	86	nM	IC50	J Med Chem (2014) 57: 3053-3074 [PMID:24617858]
ChEMBL	Binding affinity to DPP2 (unknown origin)	B	7.07	pIC50	86	nM	IC50	Medchemcomm (2014) 5: 1700-1707
ChEMBL	Inhibition of human DPP2	B	7.52	pIC50	30	nM	IC50	Bioorg Med Chem Lett (2007) 17: 507-510 [PMID:17055271]
ChEMBL	Compound was tested in vitro for inhibition of Dipeptidylpeptidase II	B	7.82	pIC50	15	nM	IC50	J Med Chem (1996) 39: 2087-2094 [PMID:8642568]
ChEMBL	EnPlex (purified enzyme activity assay): This assay is described in Bachovchin et al. Nature Chemical Biology 10, 656-663 (2014). Briefly, purified enzymes are coupled to Luminex microspheres, with a different bead color for each enzyme. Multiplexed bead complexes are incubated with a compound before being treated with a biotinylated activity-based probe and a streptavidin R-phycoerythrin conjugate (SAPE). The mixtures are scanned on a Luminex flow cytometer, where one laser detects the bead color (enzyme identity) and a second laser detects the R-phycoerythrin signal (enzyme activity). The enzyme concentration is calculated assuming 100% of the protein was coupled to the beads.	B	8.09	pIC50	8.2	nM	IC50	US-11559537-B2. Combination therapies using caspase-1 dependent anticancer agents and PGE2 antagonists (2023)
dipeptidyl peptidase 4/Dipeptidyl peptidase 4 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL284] [GtoPdb: 1612] [UniProtKB: P27487]
ChEMBL	Inhibitory activity of compound against Dipeptidylpeptidase IV (DPP IV)	B	5.7	pKi	2000	nM	Ki	J Med Chem (2003) 46: 5005-5014 [PMID:14584950]
ChEMBL	Inhibition of Dipeptidyl Peptidase IV	B	8.7	pKi	2	nM	Ki	Bioorg Med Chem Lett (2002) 12: 2825-2828 [PMID:12270155]
ChEMBL	Binding affinity for dipeptidylpeptidase IV	B	8.7	pKi	2	nM	Ki	J Med Chem (2004) 47: 4135-4141 [PMID:15293982]
GtoPdb	-	-	9.74	pKi	0.18	nM	Ki	J Med Chem (2008) 51: 6005-13 [PMID:18783201]
ChEMBL	Inhibition of human placental DPP4	B	9.74	pKi	0.18	nM	Ki	J Med Chem (2007) 50: 2391-2398 [PMID:17458948]
ChEMBL	Inhibition of human placental DPP4	B	9.74	pKi	0.18	nM	Ki	J Med Chem (2008) 51: 6005-6013 [PMID:18783201]
ChEMBL	Binding affinity to DPP-4 (unknown origin) assessed as inhibition constant	B	9.74	pKi	0.18	nM	Ki	Bioorg Med Chem (2022) 63: 116748-116748 [PMID:35453036]
ChEMBL	DPPIV Enzymatic Activity Assay: DPPIV enzymatic activity assay. To assay baseline dipeptidyl peptidase-4 (DPPIV) activity, 40 ng of recombinant human DPPIV (rhDPPIV) (R&S system, #1180-SE) or 40 ng of recombinant mouse DPPIV (rmDPPIV) (R&S system, #954-SE) was incubated with 400 μM of H-Gly-Pro-pNA substrate (BACHEM, #L-1880) in a DPPIV assay buffer (25 mM Tris, pH 8.3) for 30 min at 37° C. protected from the light in 96-well black plates (Nunc, #237108). To assay DPPIV inhibition by test compounds, test compounds were pre-incubated with the enzyme for 15 min at 37° C. before starting the reaction by substrate addition in 96-well black plates (Nunc, #237108). Para-nitroaniline (pNA) release was detected by measuring absorbance at 405 nm using a Multifunction Microplate Reader (Synergy 4, Biotek). All measurements were carried out in triplicate. Val-boroPro, a non-specific prolyl peptidase inhibitor, was used as a positive control.	B	5	pIC50	>10000	nM	IC50	US-11504364-B2. Inhibitors of fibroblast activation protein (2022)
ChEMBL	DPPIV Enzymatic Activity Assay: DPPIV enzymatic activity assay. To assay baseline dipeptidyl peptidase-4 (DPPIV) activity, 40 ng of recombinant human DPPIV (rhDPPIV) (R&S system, #1180-SE) or 40 ng of recombinant mouse DPPIV (rmDPPIV) (R&S system, #954-SE) was incubated with 400 μM of H-Gly-Pro-pNA substrate (BACHEM, #L-1880) in a DPPIV assay buffer (25 mM Tris, pH 8.3) for 30 min at 37° C. protected from the light in 96-well black plates (Nunc, #237108). To assay DPPIV inhibition by test compounds, test compounds were pre-incubated with the enzyme for 15 min at 37° C. before starting the reaction by substrate addition in 96-well black plates (Nunc, #237108). Para-nitroaniline (pNA) release was detected by measuring absorbance at 405 nm using a Multifunction Microplate Reader (Synergy 4, Biotek). All measurements were carried out in triplicate. Val-boroPro, a non-specific prolyl peptidase inhibitor, was used as a positive control.	B	5	pIC50	>10000	nM	IC50	US-11504364-B2. Inhibitors of fibroblast activation protein (2022)
ChEMBL	Inhibition of human placental DPP4 at pH 8	B	5.92	pIC50	1200	nM	IC50	J Med Chem (2007) 50: 2391-2398 [PMID:17458948]
ChEMBL	Inhibition Assay: The inhibitor solution is prepared by dissolving 3-5 mg of inhibitor in pH 2 solution (0.01 N HCl), such that the concentration of the solution is equal to 1 mg/10 mL. A 10 μL sample of this solution is then added to 990 μL of pH 8 buffer (0.1 M HEPES, 0.14 M NaCl), and the solution is allowed to stand at room temperature overnight.The enzyme solution is prepared by diluting 20 μL of DPIV (concentration 2.5 nM) into 40 mL of pH 8 buffer.The substrate solution is prepared by dissolving 2.0 mg of L-alanyl-L-proline-para-nitroanilide into 20 mL of pH 8 buffer.250 μL of enzyme solution is added to well #B1 to #H1, #A2 to #H2, and #A3 to #H3 of a 96 well plate, while well #A1 receives 250 μL of pH 8 buffer instead of enzyme solution. 904 of pH 8 buffer is then added to column 5 (from well #A5 to #H5).	B	5.92	pIC50	1200	nM	IC50	US-8933056-B2. Soft protease inhibitors and pro-soft forms thereof (2015)
ChEMBL	In Vitro Inhibition Assay: EnzymesRecombinant human DPPIV (R&D Systems, Cat. No. 1180-SE)Recombinant human DPP8 (Enzo Life Sciences, Cat. No. BML-SE527)Recombinant human DPP9 (R&D Systems, Cat. No. 5419-SE)Recombinant human DPPII (R&D Systems, Cat. No. 3438-SE)Recombinant human FAP (R&D Systems, Cat. No. 3715-SE)Recombinant human PREP (R&D Systems, Cat. No. 4308-SE)Assay Buffers25 mM Tris, pH 8.0 (DPPIV and DPP9)50 mM Tris, pH 7.5 (DPP8)25 mM MES, pH 6.0 (DPPII)50 mM Tris, 140 mM NaCl, pH 7.5 (FAP)25 mM Tris, 0.25 M NaCl, pH 7.5 (PREP)Substrates4000× substrate solution (100 mM Gly-Pro-AMC (VWR, Cat. No. 100042-646) in DMSO, DPPIV, DPP8 and DPP9)4000× substrate solution (100 mM Lys-Pro-AMC (Bachem, Cat. No. I-1745) in DMSO, DPPII)100× substrate solution (2.5 mM Z-Gly-Pro-AMC (VWR, Cat. No. I-1145.0050BA) in DMSO, FAP and PREP)General MaterialsCompound96-well black clear-bottom plates (Costar, Cat. No. 3603)InstrumentationPlate shakerMolecular Devices SpectraMax® M2e microplate readerProtocol1. To prepare the compound for the assay, dissolve it in either DMSO or, if cyclization is suspected, in pH 2.0 water (0.01 N HCl) to a final concentration of 100 mM. For pH 2.0 stocks, incubate at room temperature for a minimum of four hours and up to overnight. From this, prepare a 1 mM stock at pH 7.4 in 50 mM Tris. If the inhibitor is insoluble at this concentration, dilute the 100 mM stock 1:10 to 10 mM. Using this stock, prepare a 0.1 mM stock as described above.2. Prepare a dilution plate for the compound stocks to be tested. Add the 0.1 and/or 1 mM stocks prepared previously to row A of a 96-well plate. From this, perform 1:10 serial dilutions into the appropriate assay buffer down the columns as shown below:3. Prepare 20× substrate solution by diluting the DMSO stocks into the appropriate assay buffer.4. Dilute the enzymes into their appropriate assay buffers. The dilution factor is lot dependent and must be determined prior to performing the assay. The final enzyme concentrations should be 0.1, 0.8, 0.4, 0.2, 1.2, and 0.6 nM for DPPIV, 8, 9, II, FAP and PREP respectively. Add 180 μL to each well needed in columns 2-10.5. Add 20 μL of the compound of interest from the dilution plate prepared in step 2 to columns 2-10 of the assay plate where appropriate. Each sample should be tested in triplicate. Allow this to incubate for 10 minutes at room temperature, shaking the plate for the first two minutes.6. Add 10 μL of 20× substrate prepared in step 3 to each well and allow this to incubate for 15 minutes at room temperature, shaking the plate for the first two minutes.	B	6.15	pIC50	700	nM	IC50	US-11096924-B2. Combination therapies using immuno-dash inhibitors and PGE2 antagonists (2021)
ChEMBL	Inhibition of DDP4 at pH 7.4 preincubated for 10 mins	B	6.33	pIC50	470	nM	IC50	J Med Chem (2011) 54: 2022-2028 [PMID:21388136]
ChEMBL	In vitro for inhibition of Dipeptidylpeptidase IV.	B	7.59	pIC50	26	nM	IC50	J Med Chem (1996) 39: 2087-2094 [PMID:8642568]
ChEMBL	Binding affinity to DPP4 (unknown origin)	B	7.66	pIC50	22	nM	IC50	Medchemcomm (2014) 5: 1700-1707
ChEMBL	Inhibition of human seminal plasma DPP4 assessed as pNA release from Gly-Pro-p-nitroanilide substrate pre-incubated with enzyme for 15 min prior to substrate addition by fluorescence technique	B	7.66	pIC50	22	nM	IC50	Bioorg Med Chem Lett (2012) 22: 3412-3417 [PMID:22525314]
ChEMBL	Inhibition of DPP4 in human seminal plasma using Gly-Pro-p-nitroanilide as substrate incubated for 15 mins prior to substrate addition	B	7.66	pIC50	22	nM	IC50	ACS Med Chem Lett (2013) 4: 491-496 [PMID:24900696]
ChEMBL	Inhibition of DPP4 purified from human seminal plasma using Gly-Pro-p-nitroanilide as substrate by spectrophotometry	B	7.66	pIC50	22	nM	IC50	J Med Chem (2014) 57: 3053-3074 [PMID:24617858]
ChEMBL	Inhibition of DPP4 (unknown origin)	B	7.66	pIC50	22	nM	IC50	J Med Chem (2019) 62: 7874-7884 [PMID:31393718]
ChEMBL	Inhibition of DPP4 (unknown origin) using AMC substrate by fluorometric assay	B	7.66	pIC50	22	nM	IC50	Bioorg Med Chem Lett (2021) 37: 127846-127846 [PMID:33571650]
ChEMBL	Inhibition of DPP4 (unknown origin)	B	7.66	pIC50	22	nM	IC50	Eur J Med Chem (2022) 240: 114543-114543 [PMID:35797897]
ChEMBL	Inhibition Assay: The inhibitor solution is prepared by dissolving 3-5 mg of inhibitor in pH 2 solution (0.01 N HCl), such that the concentration of the solution is equal to 1 mg/10 mL. A 10 μL sample of this solution is then added to 990 μL of pH 8 buffer (0.1 M HEPES, 0.14 M NaCl), and the solution is allowed to stand at room temperature overnight.The enzyme solution is prepared by diluting 20 μL of DPIV (concentration 2.5 nM) into 40 mL of pH 8 buffer.The substrate solution is prepared by dissolving 2.0 mg of L-alanyl-L-proline-para-nitroanilide into 20 mL of pH 8 buffer.250 μL of enzyme solution is added to well #B1 to #H1, #A2 to #H2, and #A3 to #H3 of a 96 well plate, while well #A1 receives 250 μL of pH 8 buffer instead of enzyme solution. 904 of pH 8 buffer is then added to column 5 (from well #A5 to #H5).	B	8.77	pIC50	1.7	nM	IC50	US-8933056-B2. Soft protease inhibitors and pro-soft forms thereof (2015)
ChEMBL	Inhibition of human placental DPP4 at pH 2	B	8.8	pIC50	1.6	nM	IC50	J Med Chem (2007) 50: 2391-2398 [PMID:17458948]
ChEMBL	Inhibition of DDP4 at pH 2 preincubated for 10 mins	B	8.82	pIC50	1.5	nM	IC50	J Med Chem (2011) 54: 2022-2028 [PMID:21388136]
ChEMBL	Inhibition of human DPP4 using H-Gly-Pro-AMC as substrate preincubated for 10 mins prior to substrate addition by fluorescence assay	B	9.05	pIC50	0.9	nM	IC50	J Med Chem (2013) 56: 3467-3477 [PMID:23594271]
ChEMBL	EnPlex (purified enzyme activity assay): This assay is described in Bachovchin et al. Nature Chemical Biology 10, 656-663 (2014). Briefly, purified enzymes are coupled to Luminex microspheres, with a different bead color for each enzyme. Multiplexed bead complexes are incubated with a compound before being treated with a biotinylated activity-based probe and a streptavidin R-phycoerythrin conjugate (SAPE). The mixtures are scanned on a Luminex flow cytometer, where one laser detects the bead color (enzyme identity) and a second laser detects the R-phycoerythrin signal (enzyme activity). The enzyme concentration is calculated assuming 100% of the protein was coupled to the beads.	B	9.15	pIC50	0.7	nM	IC50	US-11559537-B2. Combination therapies using caspase-1 dependent anticancer agents and PGE2 antagonists (2023)
ChEMBL	Inhibition of human DPP4	B	10	pIC50	0.1	nM	IC50	Bioorg Med Chem Lett (2007) 17: 507-510 [PMID:17055271]
dipeptidyl peptidase 4/Dipeptidyl peptidase 4 in Mouse (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL3883] [GtoPdb: 1612] [UniProtKB: P28843]
ChEMBL	DPPIV Enzymatic Activity Assay: DPPIV enzymatic activity assay. To assay baseline dipeptidyl peptidase-4 (DPPIV) activity, 40 ng of recombinant human DPPIV (rhDPPIV) (R&S system, #1180-SE) or 40 ng of recombinant mouse DPPIV (rmDPPIV) (R&S system, #954-SE) was incubated with 400 μM of H-Gly-Pro-pNA substrate (BACHEM, #L-1880) in a DPPIV assay buffer (25 mM Tris, pH 8.3) for 30 min at 37° C. protected from the light in 96-well black plates (Nunc, #237108). To assay DPPIV inhibition by test compounds, test compounds were pre-incubated with the enzyme for 15 min at 37° C. before starting the reaction by substrate addition in 96-well black plates (Nunc, #237108). Para-nitroaniline (pNA) release was detected by measuring absorbance at 405 nm using a Multifunction Microplate Reader (Synergy 4, Biotek). All measurements were carried out in triplicate. Val-boroPro, a non-specific prolyl peptidase inhibitor, was used as a positive control.	B	5	pIC50	>10000	nM	IC50	US-11504364-B2. Inhibitors of fibroblast activation protein (2022)
dipeptidyl peptidase 8/Dipeptidyl peptidase 8 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4657] [GtoPdb: 2356] [UniProtKB: Q6V1X1]
GtoPdb	-	-	8.82	pKi	1.5	nM	Ki	J Med Chem (2008) 51: 6005-13 [PMID:18783201]
ChEMBL	Inhibition of human recombinant DPP8 expressed in HEK293T cells	B	8.82	pKi	1.5	nM	Ki	J Med Chem (2008) 51: 6005-6013 [PMID:18783201]
ChEMBL	In Vitro Inhibition Assay: EnzymesRecombinant human DPPIV (R&D Systems, Cat. No. 1180-SE)Recombinant human DPP8 (Enzo Life Sciences, Cat. No. BML-SE527)Recombinant human DPP9 (R&D Systems, Cat. No. 5419-SE)Recombinant human DPPII (R&D Systems, Cat. No. 3438-SE)Recombinant human FAP (R&D Systems, Cat. No. 3715-SE)Recombinant human PREP (R&D Systems, Cat. No. 4308-SE)Assay Buffers25 mM Tris, pH 8.0 (DPPIV and DPP9)50 mM Tris, pH 7.5 (DPP8)25 mM MES, pH 6.0 (DPPII)50 mM Tris, 140 mM NaCl, pH 7.5 (FAP)25 mM Tris, 0.25 M NaCl, pH 7.5 (PREP)Substrates4000× substrate solution (100 mM Gly-Pro-AMC (VWR, Cat. No. 100042-646) in DMSO, DPPIV, DPP8 and DPP9)4000× substrate solution (100 mM Lys-Pro-AMC (Bachem, Cat. No. I-1745) in DMSO, DPPII)100× substrate solution (2.5 mM Z-Gly-Pro-AMC (VWR, Cat. No. I-1145.0050BA) in DMSO, FAP and PREP)General MaterialsCompound96-well black clear-bottom plates (Costar, Cat. No. 3603)InstrumentationPlate shakerMolecular Devices SpectraMax® M2e microplate readerProtocol1. To prepare the compound for the assay, dissolve it in either DMSO or, if cyclization is suspected, in pH 2.0 water (0.01 N HCl) to a final concentration of 100 mM. For pH 2.0 stocks, incubate at room temperature for a minimum of four hours and up to overnight. From this, prepare a 1 mM stock at pH 7.4 in 50 mM Tris. If the inhibitor is insoluble at this concentration, dilute the 100 mM stock 1:10 to 10 mM. Using this stock, prepare a 0.1 mM stock as described above.2. Prepare a dilution plate for the compound stocks to be tested. Add the 0.1 and/or 1 mM stocks prepared previously to row A of a 96-well plate. From this, perform 1:10 serial dilutions into the appropriate assay buffer down the columns as shown below:3. Prepare 20× substrate solution by diluting the DMSO stocks into the appropriate assay buffer.4. Dilute the enzymes into their appropriate assay buffers. The dilution factor is lot dependent and must be determined prior to performing the assay. The final enzyme concentrations should be 0.1, 0.8, 0.4, 0.2, 1.2, and 0.6 nM for DPPIV, 8, 9, II, FAP and PREP respectively. Add 180 μL to each well needed in columns 2-10.5. Add 20 μL of the compound of interest from the dilution plate prepared in step 2 to columns 2-10 of the assay plate where appropriate. Each sample should be tested in triplicate. Allow this to incubate for 10 minutes at room temperature, shaking the plate for the first two minutes.6. Add 10 μL of 20× substrate prepared in step 3 to each well and allow this to incubate for 15 minutes at room temperature, shaking the plate for the first two minutes.	B	5.44	pIC50	3600	nM	IC50	US-11096924-B2. Combination therapies using immuno-dash inhibitors and PGE2 antagonists (2021)
ChEMBL	Inhibition of human DPP8	B	7.77	pIC50	17	nM	IC50	Bioorg Med Chem Lett (2007) 17: 507-510 [PMID:17055271]
ChEMBL	Inhibition of human DPP8 using H-Gly-Pro-AMC as substrate preincubated for 10 mins prior to substrate addition by fluorescence assay	B	8.26	pIC50	5.5	nM	IC50	J Med Chem (2013) 56: 3467-3477 [PMID:23594271]
ChEMBL	EnPlex (purified enzyme activity assay): This assay is described in Bachovchin et al. Nature Chemical Biology 10, 656-663 (2014). Briefly, purified enzymes are coupled to Luminex microspheres, with a different bead color for each enzyme. Multiplexed bead complexes are incubated with a compound before being treated with a biotinylated activity-based probe and a streptavidin R-phycoerythrin conjugate (SAPE). The mixtures are scanned on a Luminex flow cytometer, where one laser detects the bead color (enzyme identity) and a second laser detects the R-phycoerythrin signal (enzyme activity). The enzyme concentration is calculated assuming 100% of the protein was coupled to the beads.	B	8.44	pIC50	3.6	nM	IC50	US-11559537-B2. Combination therapies using caspase-1 dependent anticancer agents and PGE2 antagonists (2023)
dipeptidyl peptidase 9/Dipeptidyl peptidase 9 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4793] [GtoPdb: 2357] [UniProtKB: Q86TI2]
ChEMBL	Inhibition of human recombinant DPP9 expressed in HEK293T cells	B	9.12	pKi	0.76	nM	Ki	J Med Chem (2008) 51: 6005-6013 [PMID:18783201]
GtoPdb	-	-	9.12	pKi	0.76	nM	Ki	J Med Chem (2008) 51: 6005-13 [PMID:18783201]
ChEMBL	DPP9 Enzymatic Activity Assay: DPP9 enzymatic activity assay. To assay baseline dipeptidyl peptidase 9 (DPP9) activity, 40 ng of recombinant human DPP9 (rhDPP9) (R&S system, #5419-SE) was incubated with 100 μM of H-Gly-Pro-AMC peptide (BACHEM, #L-1215) in a DDP9 assay buffer (50 mM HEPES, pH 8) for 30 min at 37° C. in 96-well black plates (Nunc, #237108). To assay rhDPP9 activity inhibition by test compounds, test compounds were pre-incubated with the enzyme for 15 min at 37° C. before starting the reaction by substrate addition in 96-well black plates (Nunc, #237108). 7-Amino-4-Methylcoumarin (AMC) release was detected by measuring fluorescence at Ex/Em 380/460 nm using a Multifunction Microplate Reader (Synergy 4, Biotek). All measurements were carried out in triplicate. Val-boroPro, a non-specific prolyl peptidase inhibitor, was used as a positive control.	B	5	pIC50	>10000	nM	IC50	US-11504364-B2. Inhibitors of fibroblast activation protein (2022)
ChEMBL	DPP9 Enzymatic Activity Assay: DPP9 enzymatic activity assay. To assay baseline dipeptidyl peptidase 9 (DPP9) activity, 40 ng of recombinant human DPP9 (rhDPP9) (R&S system, #5419-SE) was incubated with 100 μM of H-Gly-Pro-AMC peptide (BACHEM, #L-1215) in a DDP9 assay buffer (50 mM HEPES, pH 8) for 30 min at 37° C. in 96-well black plates (Nunc, #237108). To assay rhDPP9 activity inhibition by test compounds, test compounds were pre-incubated with the enzyme for 15 min at 37° C. before starting the reaction by substrate addition in 96-well black plates (Nunc, #237108). 7-Amino-4-Methylcoumarin (AMC) release was detected by measuring fluorescence at Ex/Em 380/460 nm using a Multifunction Microplate Reader (Synergy 4, Biotek). All measurements were carried out in triplicate. Val-boroPro, a non-specific prolyl peptidase inhibitor, was used as a positive control.	B	5	pIC50	>10000	nM	IC50	US-11504364-B2. Inhibitors of fibroblast activation protein (2022)
ChEMBL	Inhibition of human recombinant DPP9 expressed in intact HEK293T cells after 2 hrs	B	5.17	pIC50	6800	nM	IC50	J Med Chem (2008) 51: 6005-6013 [PMID:18783201]
ChEMBL	In Vitro Inhibition Assay: EnzymesRecombinant human DPPIV (R&D Systems, Cat. No. 1180-SE)Recombinant human DPP8 (Enzo Life Sciences, Cat. No. BML-SE527)Recombinant human DPP9 (R&D Systems, Cat. No. 5419-SE)Recombinant human DPPII (R&D Systems, Cat. No. 3438-SE)Recombinant human FAP (R&D Systems, Cat. No. 3715-SE)Recombinant human PREP (R&D Systems, Cat. No. 4308-SE)Assay Buffers25 mM Tris, pH 8.0 (DPPIV and DPP9)50 mM Tris, pH 7.5 (DPP8)25 mM MES, pH 6.0 (DPPII)50 mM Tris, 140 mM NaCl, pH 7.5 (FAP)25 mM Tris, 0.25 M NaCl, pH 7.5 (PREP)Substrates4000× substrate solution (100 mM Gly-Pro-AMC (VWR, Cat. No. 100042-646) in DMSO, DPPIV, DPP8 and DPP9)4000× substrate solution (100 mM Lys-Pro-AMC (Bachem, Cat. No. I-1745) in DMSO, DPPII)100× substrate solution (2.5 mM Z-Gly-Pro-AMC (VWR, Cat. No. I-1145.0050BA) in DMSO, FAP and PREP)General MaterialsCompound96-well black clear-bottom plates (Costar, Cat. No. 3603)InstrumentationPlate shakerMolecular Devices SpectraMax® M2e microplate readerProtocol1. To prepare the compound for the assay, dissolve it in either DMSO or, if cyclization is suspected, in pH 2.0 water (0.01 N HCl) to a final concentration of 100 mM. For pH 2.0 stocks, incubate at room temperature for a minimum of four hours and up to overnight. From this, prepare a 1 mM stock at pH 7.4 in 50 mM Tris. If the inhibitor is insoluble at this concentration, dilute the 100 mM stock 1:10 to 10 mM. Using this stock, prepare a 0.1 mM stock as described above.2. Prepare a dilution plate for the compound stocks to be tested. Add the 0.1 and/or 1 mM stocks prepared previously to row A of a 96-well plate. From this, perform 1:10 serial dilutions into the appropriate assay buffer down the columns as shown below:3. Prepare 20× substrate solution by diluting the DMSO stocks into the appropriate assay buffer.4. Dilute the enzymes into their appropriate assay buffers. The dilution factor is lot dependent and must be determined prior to performing the assay. The final enzyme concentrations should be 0.1, 0.8, 0.4, 0.2, 1.2, and 0.6 nM for DPPIV, 8, 9, II, FAP and PREP respectively. Add 180 μL to each well needed in columns 2-10.5. Add 20 μL of the compound of interest from the dilution plate prepared in step 2 to columns 2-10 of the assay plate where appropriate. Each sample should be tested in triplicate. Allow this to incubate for 10 minutes at room temperature, shaking the plate for the first two minutes.6. Add 10 μL of 20× substrate prepared in step 3 to each well and allow this to incubate for 15 minutes at room temperature, shaking the plate for the first two minutes.	B	5.77	pIC50	1700	nM	IC50	US-11096924-B2. Combination therapies using immuno-dash inhibitors and PGE2 antagonists (2021)
ChEMBL	Inhibition of human DPP9 using H-Gly-Pro-AMC as substrate preincubated for 10 mins prior to substrate addition by fluorescence assay	B	8.66	pIC50	2.2	nM	IC50	J Med Chem (2013) 56: 3467-3477 [PMID:23594271]
ChEMBL	Inhibition of human DPP9	B	8.7	pIC50	2	nM	IC50	Bioorg Med Chem Lett (2007) 17: 507-510 [PMID:17055271]
ChEMBL	EnPlex (purified enzyme activity assay): This assay is described in Bachovchin et al. Nature Chemical Biology 10, 656-663 (2014). Briefly, purified enzymes are coupled to Luminex microspheres, with a different bead color for each enzyme. Multiplexed bead complexes are incubated with a compound before being treated with a biotinylated activity-based probe and a streptavidin R-phycoerythrin conjugate (SAPE). The mixtures are scanned on a Luminex flow cytometer, where one laser detects the bead color (enzyme identity) and a second laser detects the R-phycoerythrin signal (enzyme activity). The enzyme concentration is calculated assuming 100% of the protein was coupled to the beads.	B	8.77	pIC50	1.7	nM	IC50	US-11559537-B2. Combination therapies using caspase-1 dependent anticancer agents and PGE2 antagonists (2023)
Presequence protease, mitochondrial in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL3124731] [UniProtKB: Q5JRX3]
ChEMBL	PREP Enzymatic Activity Assay: PREP enzymatic activity assay. To assay baseline prolyl endopeptidase (PREP) activity, 20 ng of recombinant human PREP (rhPREP) (R&S system, #4308-SE) or 20 ng of recombinant mouse PREP (rmPREP) (R&S system, #6339-SE) was incubated with 100 μM of Z-Gly-Pro-AMC peptide (BACHEM, #L-1145) in a PREP assay buffer (25 mM Tris, 250 mM NaCl, 10 mM DTT, pH 7.5) for 30 min at 37° C. protected from light in 96-well black plates (Nunc, #237108). To assay PREP activity inhibition by test compounds, test compounds were pre-incubated with the enzyme for 15 min at 37° C. before starting the reaction by substrate addition in 96-well black plates (Nunc, #237108). 7-Amino-4-Methylcoumarin (AMC) release was detected by measuring fluorescence at Ex/Em 380/460 nm using a Multifunction Microplate Reader (Synergy 4, Biotek). All measurements were carried out in triplicate. Val-boroPro, a non-specific prolyl peptidase inhibitor, was used as a positive control.	B	5.26	pIC50	5500	nM	IC50	US-11504364-B2. Inhibitors of fibroblast activation protein (2022)
ChEMBL	PREP Enzymatic Activity Assay: PREP enzymatic activity assay. To assay baseline prolyl endopeptidase (PREP) activity, 20 ng of recombinant human PREP (rhPREP) (R&S system, #4308-SE) or 20 ng of recombinant mouse PREP (rmPREP) (R&S system, #6339-SE) was incubated with 100 μM of Z-Gly-Pro-AMC peptide (BACHEM, #L-1145) in a PREP assay buffer (25 mM Tris, 250 mM NaCl, 10 mM DTT, pH 7.5) for 30 min at 37° C. protected from light in 96-well black plates (Nunc, #237108). To assay PREP activity inhibition by test compounds, test compounds were pre-incubated with the enzyme for 15 min at 37° C. before starting the reaction by substrate addition in 96-well black plates (Nunc, #237108). 7-Amino-4-Methylcoumarin (AMC) release was detected by measuring fluorescence at Ex/Em 380/460 nm using a Multifunction Microplate Reader (Synergy 4, Biotek). All measurements were carried out in triplicate. Val-boroPro, a non-specific prolyl peptidase inhibitor, was used as a positive control.	B	5.26	pIC50	5500	nM	IC50	US-11504364-B2. Inhibitors of fibroblast activation protein (2022)
prolyl endopeptidase/Prolyl endopeptidase in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL3202] [GtoPdb: 2395] [UniProtKB: P48147]
ChEMBL	Compound was tested in vitro for inhibition of Prolyl endopeptidase	B	4.64	pIC50	23000	nM	IC50	J Med Chem (1996) 39: 2087-2094 [PMID:8642568]
ChEMBL	Inhibition of prolyl oligopeptidase	B	5.1	pIC50	7960	nM	IC50	Bioorg Med Chem Lett (2007) 17: 507-510 [PMID:17055271]
ChEMBL	Inhibition of human recombinant PREP expressed in Escherichia coli using Z-Gly-Pro-p-nitroanilide as substrate incubated for 15 mins prior to substrate addition	B	6.01	pIC50	980	nM	IC50	ACS Med Chem Lett (2013) 4: 491-496 [PMID:24900696]
ChEMBL	Inhibition of recombinant human PREP purified from Escherichia coli using Z-Gly-Pro-p-nitroanilide as substrate by spectrophotometry	B	6.01	pIC50	980	nM	IC50	J Med Chem (2014) 57: 3053-3074 [PMID:24617858]
ChEMBL	Binding affinity to PREP (unknown origin)	B	6.01	pIC50	980	nM	IC50	Medchemcomm (2014) 5: 1700-1707
ChEMBL	Inhibition of POP (unknown origin)	B	6.01	pIC50	980	nM	IC50	J Med Chem (2019) 62: 7874-7884 [PMID:31393718]
ChEMBL	Inhibition of PREP (unknown origin) using AMC substrate by fluorometric assay	B	6.01	pIC50	980	nM	IC50	Bioorg Med Chem Lett (2021) 37: 127846-127846 [PMID:33571650]
ChEMBL	Inhibition of human recombinant PREP expressed in Escherichia coli assessed as pNA release from Z-Gly-Pro-p-nitroanilide pre-incubated with enzyme for 15 mins prior to substrate addition by fluorescence technique	B	6.01	pIC50	980	nM	IC50	Bioorg Med Chem Lett (2012) 22: 3412-3417 [PMID:22525314]
ChEMBL	Inhibition of human PREP using Z-Gly-Pro-AMC as substrate preincubated for 10 mins prior to substrate addition by fluorescence assay	B	7.14	pIC50	73	nM	IC50	J Med Chem (2013) 56: 3467-3477 [PMID:23594271]
fibroblast activation protein alpha/Prolyl endopeptidase FAP in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4683] [GtoPdb: 2365] [UniProtKB: Q12884]
ChEMBL	Inhibition of FAP (unknown origin)	B	7.18	pKi	66	nM	Ki	Eur J Med Chem (2022) 240: 114543-114543 [PMID:35797897]
ChEMBL	Binding affinity to FAP (unknown origin) assessed as inhibition constant	B	8.3	pKi	5	nM	Ki	Bioorg Med Chem (2022) 63: 116748-116748 [PMID:35453036]
ChEMBL	In Vitro Inhibition Assay: EnzymesRecombinant human DPPIV (R&D Systems, Cat. No. 1180-SE)Recombinant human DPP8 (Enzo Life Sciences, Cat. No. BML-SE527)Recombinant human DPP9 (R&D Systems, Cat. No. 5419-SE)Recombinant human DPPII (R&D Systems, Cat. No. 3438-SE)Recombinant human FAP (R&D Systems, Cat. No. 3715-SE)Recombinant human PREP (R&D Systems, Cat. No. 4308-SE)Assay Buffers25 mM Tris, pH 8.0 (DPPIV and DPP9)50 mM Tris, pH 7.5 (DPP8)25 mM MES, pH 6.0 (DPPII)50 mM Tris, 140 mM NaCl, pH 7.5 (FAP)25 mM Tris, 0.25 M NaCl, pH 7.5 (PREP)Substrates4000× substrate solution (100 mM Gly-Pro-AMC (VWR, Cat. No. 100042-646) in DMSO, DPPIV, DPP8 and DPP9)4000× substrate solution (100 mM Lys-Pro-AMC (Bachem, Cat. No. I-1745) in DMSO, DPPII)100× substrate solution (2.5 mM Z-Gly-Pro-AMC (VWR, Cat. No. I-1145.0050BA) in DMSO, FAP and PREP)General MaterialsCompound96-well black clear-bottom plates (Costar, Cat. No. 3603)InstrumentationPlate shakerMolecular Devices SpectraMax® M2e microplate readerProtocol1. To prepare the compound for the assay, dissolve it in either DMSO or, if cyclization is suspected, in pH 2.0 water (0.01 N HCl) to a final concentration of 100 mM. For pH 2.0 stocks, incubate at room temperature for a minimum of four hours and up to overnight. From this, prepare a 1 mM stock at pH 7.4 in 50 mM Tris. If the inhibitor is insoluble at this concentration, dilute the 100 mM stock 1:10 to 10 mM. Using this stock, prepare a 0.1 mM stock as described above.2. Prepare a dilution plate for the compound stocks to be tested. Add the 0.1 and/or 1 mM stocks prepared previously to row A of a 96-well plate. From this, perform 1:10 serial dilutions into the appropriate assay buffer down the columns as shown below:3. Prepare 20× substrate solution by diluting the DMSO stocks into the appropriate assay buffer.4. Dilute the enzymes into their appropriate assay buffers. The dilution factor is lot dependent and must be determined prior to performing the assay. The final enzyme concentrations should be 0.1, 0.8, 0.4, 0.2, 1.2, and 0.6 nM for DPPIV, 8, 9, II, FAP and PREP respectively. Add 180 μL to each well needed in columns 2-10.5. Add 20 μL of the compound of interest from the dilution plate prepared in step 2 to columns 2-10 of the assay plate where appropriate. Each sample should be tested in triplicate. Allow this to incubate for 10 minutes at room temperature, shaking the plate for the first two minutes.6. Add 10 μL of 20× substrate prepared in step 3 to each well and allow this to incubate for 15 minutes at room temperature, shaking the plate for the first two minutes.	B	4.77	pIC50	17000	nM	IC50	US-11096924-B2. Combination therapies using immuno-dash inhibitors and PGE2 antagonists (2021)
ChEMBL	Inhibition of FAPα In Vitro Enzymatic Activity Assay: FAPα enzymatic exopeptidase (dipeptidase) activity assay. To assay baseline FAPa enzymatic exopeptidase activity, 40 ng of recombinant human FAPα (rhFAPα, R&S system, #3715-SE) or 40 ng of recombinant mouse FAPα (rmFAPα, R&S system, #8647-SE) was incubated with 100 μM of Z-Gly-Pro-AMC peptide (BACHEM, #L-1145) in a FAPα assay buffer (50 mM Tris pH 7.4, 100 mM NaCl, 0.1 mg/ml bovine serum albumin) for 1 h at 37° C. protected from light in 96-well black plates (Nunc, #237108). To assay FAPα enzymatic exopeptidase activity inhibition by test compounds, all test compounds were pre-incubated with the enzyme for 15 min at 37° C. before starting the reaction by substrate addition in 96-well black plates (Nunc, #237108). 7-Amino-4-Methylcoumarin (AMC) release was detected by measuring fluorescence at Ex/Em 380/460 nm using a Multifunction Microplate Reader (Synergy 4, Biotek). All measurements were carried out in duplicate. Val-boroPro, a non-specific prolyl peptidase inhibitor, was used as a positive control.	B	5.26	pIC50	5500	nM	IC50	US-11504364-B2. Inhibitors of fibroblast activation protein (2022)
ChEMBL	Inhibition of FAPα In Vitro Enzymatic Activity Assay: FAPα enzymatic exopeptidase (dipeptidase) activity assay. To assay baseline FAPa enzymatic exopeptidase activity, 40 ng of recombinant human FAPα (rhFAPα, R&S system, #3715-SE) or 40 ng of recombinant mouse FAPα (rmFAPα, R&S system, #8647-SE) was incubated with 100 μM of Z-Gly-Pro-AMC peptide (BACHEM, #L-1145) in a FAPα assay buffer (50 mM Tris pH 7.4, 100 mM NaCl, 0.1 mg/ml bovine serum albumin) for 1 h at 37° C. protected from light in 96-well black plates (Nunc, #237108). To assay FAPα enzymatic exopeptidase activity inhibition by test compounds, all test compounds were pre-incubated with the enzyme for 15 min at 37° C. before starting the reaction by substrate addition in 96-well black plates (Nunc, #237108). 7-Amino-4-Methylcoumarin (AMC) release was detected by measuring fluorescence at Ex/Em 380/460 nm using a Multifunction Microplate Reader (Synergy 4, Biotek). All measurements were carried out in duplicate. Val-boroPro, a non-specific prolyl peptidase inhibitor, was used as a positive control.	B	5.26	pIC50	5500	nM	IC50	US-11504364-B2. Inhibitors of fibroblast activation protein (2022)
ChEMBL	Inhibition of FAP in human U87MG cells using Suc-Gly-Pro-AMC as substrate by fluorescence based assay	B	6.65	pIC50	224	nM	IC50	Bioorg Med Chem Lett (2020) 30: 127253-127253 [PMID:32527554]
ChEMBL	Binding affinity to FAP (unknown origin)	B	7.15	pIC50	70	nM	IC50	Medchemcomm (2014) 5: 1700-1707
ChEMBL	Inhibition of FAP (unknown origin) using AMC substrate by fluorometric assay	B	7.15	pIC50	70	nM	IC50	Bioorg Med Chem Lett (2021) 37: 127846-127846 [PMID:33571650]
ChEMBL	Inhibition of FAP (unknown origin)	B	7.18	pIC50	66	nM	IC50	J Med Chem (2019) 62: 7874-7884 [PMID:31393718]
ChEMBL	Inhibition of human FAP	B	7.62	pIC50	24	nM	IC50	Bioorg Med Chem Lett (2007) 17: 507-510 [PMID:17055271]
ChEMBL	EnPlex (purified enzyme activity assay): This assay is described in Bachovchin et al. Nature Chemical Biology 10, 656-663 (2014). Briefly, purified enzymes are coupled to Luminex microspheres, with a different bead color for each enzyme. Multiplexed bead complexes are incubated with a compound before being treated with a biotinylated activity-based probe and a streptavidin R-phycoerythrin conjugate (SAPE). The mixtures are scanned on a Luminex flow cytometer, where one laser detects the bead color (enzyme identity) and a second laser detects the R-phycoerythrin signal (enzyme activity). The enzyme concentration is calculated assuming 100% of the protein was coupled to the beads.	B	7.77	pIC50	17	nM	IC50	US-11559537-B2. Combination therapies using caspase-1 dependent anticancer agents and PGE2 antagonists (2023)
ChEMBL	Inhibition of human FAP using Z-Gly-Pro-AMC as substrate preincubated for 10 mins prior to substrate addition by fluorescence assay	B	7.92	pIC50	12	nM	IC50	J Med Chem (2013) 56: 3467-3477 [PMID:23594271]
fibroblast activation protein alpha/Prolyl endopeptidase FAP in Mouse (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL5769] [GtoPdb: 2365] [UniProtKB: P97321]
ChEMBL	Inhibition of mouse recombinant FAP expressed in HEK293 cells using Ala-Pro-p-nitroanilide as substrate incubated for 15 mins prior to substrate addition	B	7.15	pIC50	70	nM	IC50	ACS Med Chem Lett (2013) 4: 491-496 [PMID:24900696]
ChEMBL	Inhibition of mouse recombinant FAP expressed in HEK293 cells assessed as pNA release from Ala-Pro-p-nitroanilide pre-incubated with enzyme for 15 mins prior to substrate addition by fluorescence technique	B	7.18	pIC50	66	nM	IC50	Bioorg Med Chem Lett (2012) 22: 3412-3417 [PMID:22525314]
ChEMBL	Inhibition of recombinant mouse FAP purified from HEK293 cell supernatant using Ala-Pro-p-nitroanilide as substrate by spectrophotometry	B	7.18	pIC50	66	nM	IC50	J Med Chem (2014) 57: 3053-3074 [PMID:24617858]
peptidase D/Xaa-Pro dipeptidase in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4185] [GtoPdb: 2389] [UniProtKB: P12955]
ChEMBL	Inhibition of recombinant human PEPD using Ala-Pro as substrate assessed as alanine release by measuring increase in fluorescense signal by fluorescence based analysis	B	5	pIC50	~10000	nM	IC50	J Med Chem (2023) 66: 2589-2607 [PMID:36724486]

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]