Summary assay for compounds activating TNAP performed in a luminescent assay
Assay Providers: Drs. Jose Luis Millan and Eduard Sergienko, Sanford-Burnham Medical Research Institute, San Diego, CA. ..more
Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG)
Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA)
Network: NIH Molecular Libraries Screening Centers Network (MLSCN)
Grant Proposal Number: 1R03 MH082385-01
Assay Providers: Drs. Jose Luis Millan and Eduard Sergienko, Sanford-Burnham Medical Research Institute, San Diego, CA.
Alkaline phosphatases (EC 184.108.40.206) (APs) catalyze the hydrolysis of phosphomonoesters, releasing phosphate and alcohol. APs are dimeric enzymes found in the most organisms. In human, four isozymes of APs have been identified. Three isozymes are tissue-specific and the fourth one is tissue-non specific, named TNAP. TNAP deficiency is associated with defective bone mineralization in the form of rickets and osteomalacia. Therefore, there is therapeutic potential of modulating TNAP activity.
The goal of this HTS is to identify novel and specific activators of TNAP. The only known to date class of alkaline phosphatases activators are amino-containing alcohols, such as diethanolamine (DEA), that act as phosphoacceptor substrate and exhibit its effect in high-mM concentration range. Compounds with a similar mode of action are expected to demonstrate diminished stimulating potential if tested in the presence of DEA. Therefore, for detection of compounds with diverse mode of action, the HTS assay was optimized and HTS campaigns were performed in the presence and in the absence of DEA. The current AID reports a set of the data from screening in the absence of DEA or any other phosphate acceptor.
AIDs 518, 813, 1001, 1136, 1227, 1450 and 1659 were carried out to select probe molecules SID48410176(ML088) and SID56373763(ML087).
This AID is used to summarize the results of this MLPCN project. Probe molecules are defined as the positives of this assay and assigned a score of 100
Categorized Comment - additional comments and annotations
From MLP Probe Report:
Probe count: 2
MLP Probe ML# for probe 1: ML088
PubChem Substance ID (SID) for probe 1: 48410176
PubChem Compound ID (CID) for probe 1: 704205
Probe type for probe 1: Activator
IC50/EC50 (nM) for probe 1: 6190
Target for probe 1: TNAP (gi: 116734717)
NCBI Book chapter link for probe 1: http://www.ncbi.nlm.nih.gov/books/NBK47339/ (ID: 2358390)
Grant number for probe 1: MH082385-01
MLP Probe ML# for probe 2: ML087
PubChem Substance ID (SID) for probe 2: 56373763
PubChem Compound ID (CID) for probe 2: 25199559
Probe type for probe 2: Activator
IC50/EC50 (nM) for probe 2: 199
Target for probe 2: TNAP (gi: 116734717)
NCBI Book chapter title for probe 1: HTS identification of compounds activating TNAP at an intermediate concentration of phosphate acceptor detected in luminescent assay
Data Table (Concise)