qHTS of PTHR Inhibitors: Primary Screen
The goal of the proposed studies is to discover new compounds that act as potent agonists on the parathyroid hormone receptor type-1 (PTHR1). The PTHR1 plays critical roles in maintaining blood calcium and phosphate levels, as well as in controlling bone remodeling. PTHR1 agonists could thus be useful for treating diseases such as hypoparathyroidism and osteoporosis. In the former disease, an more ..
BioActive Compounds: 308
The goal of the proposed studies is to discover new compounds that act as potent agonists on the parathyroid hormone receptor type-1 (PTHR1). The PTHR1 plays critical roles in maintaining blood calcium and phosphate levels, as well as in controlling bone remodeling. PTHR1 agonists could thus be useful for treating diseases such as hypoparathyroidism and osteoporosis. In the former disease, an agonist drug would help maintain normal blood calcium and phosphate levels under conditions of parathyroid hormone deficiency. In the latter disease, an agonist drug would stimulate new bone formation and thus reduce the risk of fracture, under conditions of bone fragility, as occurs in aging.
To that end, we performed a quantitative high-throughput screening (qHTS) of the Molecular Libraries Probe Centers Network (MLPCN) compound libraries at the NIH Chemical Genomics Center (NCGC) for the parathyroid hormone receptor type-1 (PTHR1) using a unique HEK-293-derived cell line (GP2) that expresses the PTHR1 and a glosensor cAMP reporter construct.
NIH Molecular Libraries Probe Production Centers Network [MLPCN]
NIH Chemical Genomics Center [NCGC]
MLPCN Grant: DK011794
Assay Submitter (PI): Thomas Gardella, Massachusetts General Hospital
4 microL are dispensed into a 1536-well micro plate to yield 3,000 cells/well. The plates are then incubated overnight, after which 4 microL of 0.5 mM of BIoTium Luciferin plus 0.5 mM IBMX are added. They are then incubated at 37 degree for one hour before 23 nL of compounds are added through pinning and then incubated at 37 degree again for another hour. After the plates are kept at room temperature for 15 min before they are read on the ViewLux to measure luminescence.
1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent activators are ranked higher than compounds that showed apparent inhibition.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.
Categorized Comment - additional comments and annotations
* Activity Concentration. ** Test Concentration.
Data Table (Concise)