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BioAssay: AID 2593

qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Summary

Glucocerebrosidase (GC) catalyzes the hydrolysis of beta-glucocerebroside to glucose and ceramide in lysosomes. Mutations in the glucocerebrosidase gene result in Gaucher disease, an autosomal recessive lysosomal storage disorder. Many of the mutations encountered in patients with Gaucher disease are missense alterations that may cause misfolding, decreased stability and/or mistrafficking of this more ..
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 Tested Compounds
 Tested Compounds
All(2)
 
 
Probe(2)
 
 
Active(2)
 
 
 Tested Substances
 Tested Substances
All(2)
 
 
Probe(2)
 
 
Active(2)
 
 
AID: 2593
Data Source: NCGC (GCNS634)
BioAssay Type: Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2010-03-17
Modify Date: 2010-11-03

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: Chemical Probe: 2    Active: 2
Related Experiments
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AIDNameTypeComment
2101qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher DiseaseConfirmatorydepositor-specified cross reference: Primary screen against N370S GC from tissue homogenate.
2577qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Alpha-Glucosidase CounterscreenConfirmatorydepositor-specified cross reference: Counterscreen against alpha-glucosidase.
2578qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Alpha-Galactosidase CounterscreenConfirmatorydepositor-specified cross reference: Counterscreen against alpha-galactosidase.
2587qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Chaperone Activity in Gauche Fibroblasts After Multi-day Incubation with CompoundConfirmatorydepositor-specified cross reference: Immunostaining of N370S GC containing fibroblast lysosomes.
2588qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Activity in Non-Mutant Spleen Homogenate Using a Red Fluorescent SubstrateConfirmatorydepositor-specified cross reference: Confirmation screen against wildtype GC from tissue homogenate using red probe.
2589qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Chaperone Activity in Non-Gauche Fibroblasts After Multi-day Incubation with CompoundConfirmatorydepositor-specified cross reference: Immunostaining of wildtype GC containing fibroblast lysosomes.
2590qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Primary Screen ConfirmationConfirmatorydepositor-specified cross reference: Confirmation screen against N370S GC from tissue homogenate.
2592qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Activity in Non-Mutant Spleen HomogenateConfirmatorydepositor-specified cross reference: Confirmation screen against wildtype GC from tissue homogenate.
2595qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Purified Non-mutant GlucocerebrosidaseConfirmatorydepositor-specified cross reference: Confirmation screen against purified, wildtype GC.
2596qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Purified N370S Glucocerebrosidase Cleavage of GlucosylceramideConfirmatorydepositor-specified cross reference: Confirmation screen against purified, N370S GC using natural substrate.
2597qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Purified N370S GlucocerebrosidaseConfirmatorydepositor-specified cross reference: Confirmation screen against purified, N370S GC.
2613qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Activity in N370S Spleen Homogenate Using a Red Fluorescent SubstrateConfirmatorydepositor-specified cross reference: Confirmation screen against N370S GC from tissue homogenate using red probe.
2671qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Hit ValidationConfirmatorydepositor-specified cross reference
488834qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Alpha-Glucosidase Counterscreen for Probe SARConfirmatorydepositor-specified cross reference: Counterscreen against alpha-glucosidase.
488845qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Primary Screen Confirmation Using LC/MSConfirmatorydepositor-specified cross reference: Confirmation screen against N370S GC from tissue homogenate with alternate LC/MS detection/readout.
488846qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Alpha-Galactosidase Counterscreen for Probe SARConfirmatorydepositor-specified cross reference: Counterscreen against alpha-galactosidase.
488849qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Activity in Non-Mutant Spleen Confirmation for Probe SARConfirmatorydepositor-specified cross reference: Confirmation screen against wildtype GC from tissue homogenate.
488850qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Purified N370S Glucocerebrosidase Confirmation for Probe SARConfirmatorydepositor-specified cross reference: Confirmation screen against purified, N370S GC.
488851qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Purified Non-mutant Confirmation for Probe SARConfirmatorydepositor-specified cross reference: Confirmation screen against purified, wildtype GC.
488852qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Activity in N370S Spleen Homogenate Confirmation Using Alternate Substrate for Probe SARConfirmatorydepositor-specified cross reference: Confirmation screen against N370S GC from tissue homogenate using red probe.
488853qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Activity in Non-Mutant Spleen Using Alternate Substrate Confirmation for Probe SARConfirmatorydepositor-specified cross reference: Confirmation screen against wildtype GC from tissue homogenate using red probe.
488854qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Confirmation for Probe SARConfirmatorydepositor-specified cross reference: Confirmation screen against N370S GC from tissue homogenate.
504745Inhibitors of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Efflux Ratio ProfilingOtherdepositor-specified cross reference
504746Inhibitors of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Metabolic Stability Profile with NADPHOtherdepositor-specified cross reference
504747Inhibitors of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Caco-2 PermeabilityOtherdepositor-specified cross reference
504748Inhibitors of N370S glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Metabolic Stability ProfileOtherdepositor-specified cross reference
588853qHTS for Activators of Human Glucocerebrosidase as a Potential Chaperone Treatment of Gaucher Disease: Fibroblast TranslocationOtherdepositor-specified cross reference
Description:
NIH Chemical Genomics Center [NCGC]
NIH Molecular Libraries Probe Production Centers Network [MLPCN]

MLPCN Grant: MH086442-01
Assay Submitter (PI): Wei Zheng

Glucocerebrosidase (GC) catalyzes the hydrolysis of beta-glucocerebroside to glucose and ceramide in lysosomes. Mutations in the glucocerebrosidase gene result in Gaucher disease, an autosomal recessive lysosomal storage disorder. Many of the mutations encountered in patients with Gaucher disease are missense alterations that may cause misfolding, decreased stability and/or mistrafficking of this lysosomal protein. Some GC inhibitors have been shown to act as chemical chaperones, stabilizing the conformation of mutant proteins and thus restoring their function. These inhibitors include iminosugar analogs and three non-iminosugar inhibitors identified from a previous HTS of 62,000 compounds with the wild type recombinant enzyme. However, the enhancement of enzyme activity by the chaperone action of an enzyme inhibitor must be balanced against the direct inhibition of the enzyme. An enzyme activator could also function as a chaperone by binding to the enzyme and helping to correct its misfolding and mistrafficking. While activators for GC have not yet been identified, they may have better therapeutic potential than inhibitors. Therefore the discovery and development of chemical activators may provide a new strategy for the chaperone therapy.

We have optimized a new assay using N370S mutant GC derived from the spleen of a Gaucher patient. The new assay differs significantly from the previous screen because the N370S mutant enzyme is used instead of wildtype GC and an enzyme preparation derived from patient tissue instead of the purified recombinant GC that should have the physiologically relevant subunit/subunits and cofactors. In addition, the MLPCN compound library has expanded, which increases the chance of finding new and better probes.
Protocol
See non-summary AIDS for detailed assay protocols.
Comment
This summary is written for the purposes of summarizing the probe activities from the project. MLSCN probes are given a score of 100. Molecules in the prior art are given a score of 80. Other, less active molecules in the same chemical series as the probe molecules are given a score of 50. Inactive analogues from these series are given a score of 0. ML155 (SID 85267237) and ML156 (SID 89449177) have been declared as probes on this project.
Categorized Comment - additional comments and annotations
From MLP Probe Report:
Probe count: 4
MLP Probe ML# for probe 1: ML155
PubChem Substance ID (SID) for probe 1: 85267237
PubChem Compound ID (CID) for probe 1: 40225210
Probe type for probe 1: Modulator
IC50/EC50 (nM) for probe 1: 330
Target for probe 1: N370S Glucocerebrosidase (gi: 496369)
Disease relevance for probe 1: Gaucher Disease Rare Disease
Anti-target for probe 1: Alphaglucosidase
Fold selectivity for probe 1: >100
NCBI Book chapter link for probe 1: http://www.ncbi.nlm.nih.gov/books/NBK63601/ (ID: 2580478)
Grant number for probe 1: MH086442-01
MLP Probe ML# for probe 2: ML156
PubChem Substance ID (SID) for probe 2: 89449177
PubChem Compound ID (CID) for probe 2: 9893924
Probe type for probe 2: Modulator
IC50/EC50 (nM) for probe 2: 580
Target for probe 2: N370S Glucocerebrosidase (gi: 496369)
Disease relevance for probe 2: Gaucher Disease Rare Disease
Anti-target for probe 2: Alphaglucosidase
Fold selectivity for probe 2: >100
NCBI Book chapter link for probe 2: http://www.ncbi.nlm.nih.gov/books/NBK56229/ (ID: 2509216)
Grant number for probe 2: MH086442-01
MLP Probe ML# for probe 3: ML198
PubChem Substance ID (SID) for probe 3: 99368009
PubChem Compound ID (CID) for probe 3: 46907762
Probe type for probe 3: Activator
IC50/EC50 (nM) for probe 3: 366
Target for probe 3: N370S GCase# from Tissue Homogenate (gi: 496369)
Disease relevance for probe 3: Gaucher disease
Anti-target for probe 3: Acid alpha-glucosidase, alpha- galactosidase
Fold selectivity for probe 3: >57
NCBI Book chapter link for probe 3: http://www.ncbi.nlm.nih.gov/books/NBK143537/ (ID: 3036302)
Grant number for probe 3: MH086442-01
MLP Probe ML# for probe 4: ML266
PubChem Substance ID (SID) for probe 4: 124950541
PubChem Compound ID (CID) for probe 4: 46943215
NCBI Book chapter link for probe 4: http://www.ncbi.nlm.nih.gov/books/NBK143537/ (ID: 3036302)
Grant number for probe 4: MH086442-01
PubMed Publication ID (PMID) for probe 3: 22646221
PubMed Publication ID (PMID) for probe 4: 22646221
NCBI Book chapter title for probe 1: Identification of Modulators of the N370S Mutant Form of Glucocerebrosidase as a Potential Therapy for Gaucher Disease - Chemotype 1
NCBI Book chapter title for probe 2: Identification of Modulators of the N370S Mutant Form of Glucocerebrosidase as a Potential Therapy for Gaucher Disease - Chemotype 2
NCBI Book chapter title for probe 3: Discovery, SAR, and Biological Evaluation of Non-inhibitory Chaperones of Glucocerebrosidase
NCBI Book chapter title for probe 4: Discovery, SAR, and Biological Evaluation of Non-inhibitory Chaperones of Glucocerebrosidase
Result Definitions
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1ActivityTextual description of type of activity demonstrated by compound.String
2Glucocerebrosidase PotencyConcentration of compound demonstrating half-maximal activity of compound.FloatμM
3Alpha-Glucosidase PotencyConcentration of compound demonstrating half-maximal activity of compound.FloatμM
4Alpha-Galactosidase PotencyConcentration of compound demonstrating half-maximal activity of compound.FloatμM
5Gaucher Fibroblast PotencyConcentration of compound demonstrating increased translocation of glucocerebrosidase to the lysosome of patient-derived (N370S GC) fibroblasts.FloatμM
Additional Information
Grant Number: MH086442-01

Data Table (Concise)
Data Table ( Complete ):     View Active Data    View All Data
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