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BioAssay: AID 602359

Extended Characterization of Activators of Human Muscle isoform 2 Pyruvate Kinase: Summary

The metabolism of cancer cells is altered to support rapid proliferation. Otto Warburg was first to notice that cancer cells produce lactate even in the presence of oxygen [1,2]. This altered metabolism, known as the Warburg effect, is thought to give tumor cells a selective growth advantage relative to normal cells [3]. At least part of this abnormal metabolic phenotype is thought to be due to more ..
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AID: 602359
Data Source: NCGC (PYKH000)
BioAssay Type: Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2012-03-10
Modify Date: 2012-08-01
Target
Related Experiments
Show more
AIDNameTypeProbeComment
2095qHTS Assay for Activators of Human Muscle isoform 2 Pyruvate Kinase: SummarySummary6 depositor-specified cross reference: Summary AID of hPK probe phase of project
602377qHTS for Activators of Human Muscle Isoform Pyruvate Kinase: Extended Characterization Using A Lactate Dehydrogenase (LDH) AssayConfirmatory depositor-specified cross reference
602379qHTS for Activators of Human Muscle Isoform Pyruvate Kinase: Extended Characterization against Liver Pyruvate KinaseConfirmatory depositor-specified cross reference
602381qHTS for Activators of Human Muscle Isoform 2 Pyruvate Kinase (PK): Extended Characterization using Reticulocyte PKConfirmatory depositor-specified cross reference
602383qHTS for Activators of Human Muscle Isoform 2 Pyruvate Kinase: Muscle Isoform 1 AssayConfirmatory depositor-specified cross reference
602384qHTS for Activators of Human Muscle Isoform 2 Pyruvate Kinase: Extended CharacterizationConfirmatory depositor-specified cross reference
624387qHTS for Activators of Human Muscle Isoform 2 Pyruvate Kinase: Extended Characterization using M1 for Probe 2Confirmatory depositor-specified cross reference: SAR data against M1
624388qHTS for Activators of Human Muscle Isoform 2 Pyruvate Kinase (PK): Extended Characterization using Reticulocyte PK Probe 2Confirmatory depositor-specified cross reference
624389qHTS for Activators of Human Muscle Isoform 2 Pyruvate Kinase: Extended Characterization Using A Lactate Dehydrogenase (LDH) Assay Probe 2Confirmatory depositor-specified cross reference
624390qHTS for Activators of Human Muscle Isoform Pyruvate Kinase: Extended Characterization against Liver Pyruvate Kinase Probe 2Confirmatory depositor-specified cross reference
624391qHTS for Activators of Human Muscle Isoform 2 Pyruvate Kinase: Extended Characterization for Probe 2Confirmatory depositor-specified cross reference
1540Secondary assay for Activators of Human Pyruvate Kinase M2 isoformConfirmatory same project related to Summary assay
1631qHTS Assay for Activators of Human Muscle isoform 2 Pyruvate KinaseConfirmatory same project related to Summary assay
1751Confirmation Concentration-Response Assay for Activators of Human Muscle isoform 2 Pyruvate KinaseConfirmatory same project related to Summary assay
2533Confirmation Concentration-Response Assay for Activators of Human Muscle isoform 2 Pyruvate Kinase: for Probe SARConfirmatory same project related to Summary assay
2534Secondary Concentration-Response Assay for Activators of Human Reticulocyte Pyruvate Kinase: for Probe SARConfirmatory same project related to Summary assay
2535Secondary Concentration-Response Assay for Activators of Human Liver Pyruvate Kinase: for Probe SARConfirmatory same project related to Summary assay
2536Secondary Concentration-Response Assay for Activators of Human Muscle isoform 1 Pyruvate Kinase: for Probe SARConfirmatory same project related to Summary assay
2562Secondary LDH Assay for Activators of Human Reticulocyte Pyruvate Kinase: for Probe SARConfirmatory same project related to Summary assay
2576Secondary LDH Assay for Activators of Human Pyruvate Kinase M2 isoform: for Probe SARConfirmatory same project related to Summary assay
2620Secondary LDH Assay for Activators of Human Pyruvate Kinase M1 Isoform: for Probe SARConfirmatory same project related to Summary assay
2625Secondary LDH Assay for Activators of Human Liver Pyruvate Kinase: for Probe SARConfirmatory same project related to Summary assay
2653Secondary Assay for Activators of Human Pyruvate Kinase M2 isoform - Cell Titer Glo Cytotoxicity for Probe SARConfirmatory same project related to Summary assay
Description:
The metabolism of cancer cells is altered to support rapid proliferation. Otto Warburg was first to notice that cancer cells produce lactate even in the presence of oxygen [1,2]. This altered metabolism, known as the Warburg effect, is thought to give tumor cells a selective growth advantage relative to normal cells [3]. At least part of this abnormal metabolic phenotype is thought to be due to over expression of specific enzymes such as a tumor specific isozyme of pyruvate kinase - PKM2. Pyruvate kinase operates the final rate determining step of glycolysis where the enzyme converts one molecule of phosphoenol pyruvate and ADP to pyruvate and ATP. Recent work has shown that replacement of PKM2 in cancer cells with an isoform found in typical adult tissue cells (PKM1) can relieve the Warburg effect and promote a state of metabolism characteristic of normal cells [4]. Further, tumors engineered to express only PKM1 showed delayed tumor formation in nude mouse xenografts [4]. It has also been demonstrated that PKM2 has high affinity for binding phosphotyrosine peptides that are characteristic of growth factor signaling pathways present in cancer cells [5]. The binding of phosphopeptides to PKM2 removes the allosteric activator fructose-1,6-bisphosphate which further down-regulates the activity of PKM2 and intensifies the Warburg effect. Therefore, pharmacological activators of PKM2 are thought to be an approach for altering the Warburg effect by shifting the glycolytic flux from a proliferative mode back toward normal energy production.
The primary qHTS data of human PKM2 and confirmatory data are available in PubChem (AIDs: 1631, 1751 and 2533). Follow-up of synthesized analogs was determined using the same luminescent protocol for the PK isoforms M1, L and R (PubChem AIDs for M1, L and R bioluminescent assays are 2536, 2535, and 2534, respectively). A fluorescent secondary assay that determined the activity of PK by coupling the production pyruvate to lactate dehydrogenase oxidation of NADH was also used to determine the activation potency of the synthesized analog. (PubChem AIDs: 1540 and 2576 for M2 isoform; 2625, 2620, 2562 for L, M1 and R isoforms respectively). As well, general cytotoxicity of the compounds was measured in HeLa cells using Cell-TiterGlo (Promega; PubChem AID: 2653).

A bis-sulfonamide compound (CID_650361, ML083) and a heteocycle compound (CID_654376, ML082) were identified as human PKM2 activators during the probe phase of the project. These molecules were further characterized in the extended characterization part of the project.

NIH Chemical Genomics Center [NCGC]
NIH Molecular Libraries Probe Production centers Network [MLPCN]

MLPCN Grant: MH085679
Assay Submitter (PI): Matthew G. Vander Heiden
Protocol
Please see linked AIDs for a detailed protocol.
Comment
This project is on-going and will be updated at a later point.
Categorized Comment - additional comments and annotations
From PubChem:
Assay Cell Type: HeLa
From MLP Probe Report:
Probe count: 2
MLP Probe ML# for probe 1: ML265
PubChem Substance ID (SID) for probe 1: 85267885
PubChem Compound ID (CID) for probe 1: 44246499
Probe type for probe 1: Activator
IC50/EC50 (nM) for probe 1: 92
Target for probe 1: hPyk-M2 (gi: 33286418)
Disease relevance for probe 1: Cancer
Anti-target for probe 1: hPyk-M1, -R, and -L
Fold selectivity for probe 1: >100
NCBI Book chapter link for probe 1: http://www.ncbi.nlm.nih.gov/books/NBK153222/ (ID: 3046322)
Grant number for probe 1: MH085679-01
MLP Probe ML# for probe 2: ML285
PubChem Substance ID (SID) for probe 2: 85267861
PubChem Compound ID (CID) for probe 2: 25210493
Probe type for probe 2: Inhibitor
IC50/EC50 (nM) for probe 2: 82
Target for probe 2: Human Pyruvate Kinase M2 (gi: 33286418)
Disease relevance for probe 2: Cancer
Anti-target for probe 2: hPyk-M1, -R, and -L
Fold selectivity for probe 2: 100
NCBI Book chapter link for probe 2: http://www.ncbi.nlm.nih.gov/books/NBK148850/ (ID: 3044006)
Grant number for probe 2: MH085679-01
NCBI Book chapter title for probe 1: ML265: A potent PKM2 activator induces tetramerization and reduces tumor formation and size in a mouse xenograft model
NCBI Book chapter title for probe 2: ML285 affects reactive oxygen speciesí inhibition of pyruvate kinase M2
Additional Information
Grant Number: MH085679

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