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Assay Detail

CHEMBL5730853

Review assay metadata, readout intent, target linkage, and publication context from the same page.

Binding
In Vitro Enzymatic: Recombinant Cat D was expressed in CHO cells. The assay buffer for CathepsinD is 0.05 M citrate pH 3.5, 10% DMSO final, 5 mM CHAPS. The Cat D enzyme (9 nM) is pre-incubated for one hour with inhibitors, typically in about 1 uL of DMSO according to a serial dilution, is added thereto. The assays are effectively started by the addition of different FRET substrates (20 nM for Cat D) and the combination is incubated for one hour. The FRET assay is terminated with by addition of Tris buffer, which raises the pH to neutrality, and the fluorescence is determined. The FRET substrate is a peptide with commercially available fluorophore and quencher, on opposite sides of the BACE cleavage site. The Cat D substrate peptide sequence is based on sequence #1 of Table 1 from Gulnik et al. FEBS Letters v413 p 379-384 1997. Proteolytic cleavage of the FRET substrate releases quenching of fluorescence (Cat D excitation 500 nm and emission 580 nm).Alternatively, a Cat D assay may also be run according to the procedure described in the article, Characterization of new fluorgenic substrates for the rapid and sensitive assay of cathepsin E and cathepsin D, J. Biochem., 125:1137, 1999.
507
Total Activities
168
Compounds Tested
3
Activity Types
0
Assay Parameters

Assay Information

Assay Type Binding
Organism Homo sapiens
Confidence 9 — Direct single protein target
Curated By Autocuration

Target

Cathepsin D (CHEMBL2581)
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

Amino-dihydrothiazine and amino-dioxido dihydrothiazine compounds as beta-secretase antagonists and methods of use
(2017)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 169 4.30 4.95
kon 169 - -
k_off 169 - -

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL5884268 6 4.95
CHEMBL5957962 3 4.93
CHEMBL5928101 3 4.91
CHEMBL5955920 3 4.75
CHEMBL5974242 3 4.57
CHEMBL5907237 3 4.56
CHEMBL5779259 3 4.55
CHEMBL6007390 3 4.55
CHEMBL6015975 3 4.52
CHEMBL5973772 3 4.52
CHEMBL5752681 3 4.51
CHEMBL5852892 3 4.51
CHEMBL5977725 3 4.51
CHEMBL5809643 3 4.49
CHEMBL5772436 3 4.49
CHEMBL5806875 3 4.47
CHEMBL5829069 3 4.44
CHEMBL5892583 3 4.43
CHEMBL5934199 3 4.41
CHEMBL5979230 3 4.37
CHEMBL5921886 3 4.36
CHEMBL5766671 3 4.36
CHEMBL5790971 3 4.34
CHEMBL5980834 3 4.34
CHEMBL5757250 3 4.34
CHEMBL6055288 3 4.34
CHEMBL6007960 3 4.33
CHEMBL5924733 3 4.32
CHEMBL6057481 3 4.30
CHEMBL5869468 3 4.29

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL5884268 IC50 = 11300.0 nM 4.95
CHEMBL5957962 IC50 = 11800.0 nM 4.93
CHEMBL5928101 IC50 = 12200.0 nM 4.91
CHEMBL5955920 IC50 = 17700.0 nM 4.75
CHEMBL5974242 IC50 = 27100.0 nM 4.57
CHEMBL5907237 IC50 = 27300.0 nM 4.56
CHEMBL5779259 IC50 = 28300.0 nM 4.55
CHEMBL6007390 IC50 = 28000.0 nM 4.55
CHEMBL5973772 IC50 = 30200.0 nM 4.52
CHEMBL6015975 IC50 = 30000.0 nM 4.52
CHEMBL5852892 IC50 = 31200.0 nM 4.51
CHEMBL5977725 IC50 = 31100.0 nM 4.51
CHEMBL5752681 IC50 = 31100.0 nM 4.51
CHEMBL5809643 IC50 = 32400.0 nM 4.49
CHEMBL5772436 IC50 = 32100.0 nM 4.49
CHEMBL5806875 IC50 = 33700.0 nM 4.47
CHEMBL5829069 IC50 = 36000.0 nM 4.44
CHEMBL5892583 IC50 = 37300.0 nM 4.43
CHEMBL5934199 IC50 = 38800.0 nM 4.41
CHEMBL5979230 IC50 = 42800.0 nM 4.37
CHEMBL5766671 IC50 = 43300.0 nM 4.36
CHEMBL5921886 IC50 = 43500.0 nM 4.36
CHEMBL5980834 IC50 = 45200.0 nM 4.34
CHEMBL5757250 IC50 = 45600.0 nM 4.34
CHEMBL6055288 IC50 = 45400.0 nM 4.34
CHEMBL5790971 IC50 = 45600.0 nM 4.34
CHEMBL6007960 IC50 = 46800.0 nM 4.33
CHEMBL5924733 IC50 = 47600.0 nM 4.32
CHEMBL6057481 IC50 = 49800.0 nM 4.30
CHEMBL5947377 IC50 = 51200.0 nM 4.29
CHEMBL5869468 IC50 = 50900.0 nM 4.29
CHEMBL5967490 IC50 = 56100.0 nM 4.25
CHEMBL5830553 IC50 = 55600.0 nM 4.25
CHEMBL5947936 IC50 = 56300.0 nM 4.25
CHEMBL6025388 IC50 = 58700.0 nM 4.23
CHEMBL6053886 IC50 = 61800.0 nM 4.21
CHEMBL6039134 IC50 = 63600.0 nM 4.20
CHEMBL5805437 IC50 = 65000.0 nM 4.19
CHEMBL5773698 IC50 = 63900.0 nM 4.19
CHEMBL5927021 IC50 = 65500.0 nM 4.18
CHEMBL5775687 IC50 = 65700.0 nM 4.18
CHEMBL5746081 IC50 = 66700.0 nM 4.18
CHEMBL5959201 IC50 = 66800.0 nM 4.17
CHEMBL5762275 IC50 = 67400.0 nM 4.17
CHEMBL5752921 IC50 = 67500.0 nM 4.17
CHEMBL5919860 IC50 = 69800.0 nM 4.16
CHEMBL5951673 IC50 = 72800.0 nM 4.14
CHEMBL6005217 IC50 = 73500.0 nM 4.13
CHEMBL5840847 IC50 = 73300.0 nM 4.13
CHEMBL6004810 IC50 = 76500.0 nM 4.12