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BioAssay: AID 493210

Inhibitors of Aldehyde Dehydrogenase 1 (ALDH1A1): Followup Confirmation and Counterscreen

Aldehyde dehydrogenase 1 (ALDH1A1) catalyzes the NAD+ dependent oxidation of a variety of endogenous and exogenous aldehydes to the corresponding carboxylic acids. The enzyme is the critical step in the metabolic activation of retinoic acid, which plays essential roles as nuclear receptor ligand. Furthermore, the precursor, retinaldehyde has recently been shown to play a fundamental role in adipogenesis and obesity, which makes inhibitor development a possible target in metabolic diseases. See [1] through [4] for more information on ALDH1A1. ..more
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 Tested Compounds
 Tested Compounds
All(87)
 
 
Active(84)
 
 
Inactive(2)
 
 
Inconclusive(1)
 
 
 Tested Substances
 Tested Substances
All(89)
 
 
Active(86)
 
 
Inactive(2)
 
 
Inconclusive(1)
 
 
AID: 493210
Data Source: NCGC (ALDH677)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2011-02-14

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 84
Related Experiments
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AIDNameTypeProbeComment
1030qHTS Assay for Inhibitors of Aldehyde Dehydrogenase 1 (ALDH1A1)Confirmatory depositor-specified cross reference: qHTS Validation Assay for Inhibitors of aldehyde dehydrogenase 1 (ALDH1A1)
2407Probe Development Summary for Inhibitors of HPGD (15-Hydroxyprostaglandin Dehydrogenase)Summary3 depositor-specified cross reference: Probe Development Summary for Inhibitors of HPGD (15-Hydroxyprostaglandin Dehydrogenase)
886qHTS Assay for Inhibitors of HADH2 (Hydroxyacyl-Coenzyme A Dehydrogenase, Type II)Confirmatory same project related to Summary assay
893qHTS Assay for Inhibitors of HSD17B4, hydroxysteroid (17-beta) dehydrogenase 4Confirmatory same project related to Summary assay
894qHTS Assay for Inhibitors of HPGD (15-Hydroxyprostaglandin Dehydrogenase)Confirmatory same project related to Summary assay
2427Thermal Shift Assay for Inhibitors of HPGD (15-Hydroxyprostaglandin Dehydrogenase)Confirmatory same project related to Summary assay
2429Confirmation qHTS Assay for Inhibitors of HPGD (15-Hydroxyprostaglandin Dehydrogenase)Confirmatory same project related to Summary assay
504587Inhibitors of HPGD: Efflux Ratio ProfilingOther same project related to Summary assay
504606Inhibitors of HPGD: Aqueous Solubility ProfilingOther same project related to Summary assay
504629Inhibitors of HPGD: Caco-2 Cell Permeability ProfilingOther same project related to Summary assay
504638Inhibitors of HPGD: Mouse Liver Microsome ProfilingOther same project related to Summary assay
504844Inhibitors of HPGD: Mouse Plasma Stability ProfilingOther same project related to Summary assay
743171Extended Characterization of HPGD Inhibitors: Activity in Primary Assay (HPGD)Confirmatory same project related to Summary assay
743196Extended Characterization of HPGD Inhibitors: SummarySummary same project related to Summary assay
Description:
Aldehyde dehydrogenase 1 (ALDH1A1) catalyzes the NAD+ dependent oxidation of a variety of endogenous and exogenous aldehydes to the corresponding carboxylic acids. The enzyme is the critical step in the metabolic activation of retinoic acid, which plays essential roles as nuclear receptor ligand. Furthermore, the precursor, retinaldehyde has recently been shown to play a fundamental role in adipogenesis and obesity, which makes inhibitor development a possible target in metabolic diseases. See [1] through [4] for more information on ALDH1A1.

Inhibition of ALDH1A1 activity was screened by utilizing propionaldehyde as an electron donor and NAD+ as an electron acceptor/cofactor. An increase in the fluorescence intensity due to conversion of NAD+ to NADH was used to measure the enzyme activity. See [5] for screening methodology used.

References:
[1] Manzer R, Qamar L, Estey T, Pappa A, Petersen DR, Vasiliou V. Molecular cloning and baculovirus expression of the rabbit corneal aldehyde dehydrogenase (ALDH1A1) cDNA. DNA Cell Biol. 2003 May;22(5):329-38.

[2] Collard F, Vertommen D, Fortpied J, Duester G, Van Schaftingen E. Identification of 3-deoxyglucosone dehydrogenase as aldehyde dehydrogenase 1A1 (retinaldehyde dehydrogenase 1). Biochimie. 2007 Mar;89(3):369-73.

[3] Ziouzenkova O, Orasanu G, Sharlach M, Akiyama TE, Berger JP, Viereck J, Hamilton JA, Tang G, Dolnikowski GG, Vogel S, Duester G, Plutzky J. Retinaldehyde represses adipogenesis and diet-induced obesity. Nat Med. 2007 Jun;13(6):695-702.

[4] Molotkov A, Duester G. Genetic evidence that retinaldehyde dehydrogenase Raldh1 (Aldh1a1) functions downstream of alcohol dehydrogenase Adh1 in metabolism of retinol to retinoic acid. J Biol Chem. 2003 Sep 19;278(38):36085-90.

[5] Inglese J, Auld DS, Jadhav A, Johnson RL, Simeonov A, Yasgar A, Zheng W, Austin CP. Quantitative high-throughput screening: a titration-based approach that efficiently identifies biological activities in large chemical libraries. Proc Natl Acad Sci. 2006 Aug 1;103(31):11473-8.

NIH Chemical Genomics Center [NCGC]
Structural Genomics Consortium [SGC]
Protocol
Buffer: 100 mM Hepes pH 7.5, 0.01% Tween 20.
Reagents/Controls:
[1] Buffer in columns 3 and 4 as negative control (no enzyme).
[2] Substrate/cofactor solution: 1 mM NAD+ and 80 uM propionaldehyde (Sigma-Aldrich, St. Louis, MO) final concentrations dispensed throughout the plate.
[3] Enzyme: 50 nM ALDH1A1 final concentration in columns 1, 2, 5-48. Column 1 is neutral (100% activity).
[4] Control: Pintool transfer of control inhibitor Bay 11-7085 (Lopac B5681 ) to column 2 of all assay plates. Two-fold, 16 pt dilution in duplicate: 5.75 uM - 0.175 nM.
Assay Steps:
Three uL of enzyme were dispensed to 1536-well Greiner black solid bottom plates. Compounds (23 nL) were transferred via Kalypsys PinTool. The plates were incubated for 15 min at room temperature, and then 1 uL of substrate/cofactor solution was added to start the reaction. The plates were immediately transferred to and read twice on ViewLux High-throughput CCD imager (Perkin-Elmer) 20 minutes apart using 360 nm excitation and 450 nm emission fluorescence protocol. The fluorescence intensity difference between the second and the first time points was used to compute reaction progress.
Comment
Keywords: NIH Roadmap, MLSCN, MLPCN, MLI, MLSMR, SGC, qHTS, NCGC, ALDH1A1, aldehyde dehydrogenase 1 family, member A1, ALDH1, 5U54MH084681-02 U54 CDP
Compound Ranking:
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 inhibitors are ranked higher than compounds that showed apparent activation.
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
From ChEMBL:
Assay Type: Functional
Result Definitions
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TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1PhenotypeIndicates type of activity observed: inhibitor, activator, fluorescent, cytotoxic, inactive, or inconclusive.String
2Potency*Concentration at which compound exhibits half-maximal efficacy, AC50. Extrapolated AC50s also include the highest efficacy observed and the concentration of compound at which it was observed.FloatμM
3EfficacyMaximal efficacy of compound, reported as a percentage of control. These values are estimated based on fits of the Hill equation to the dose-response curves.Float%
4Analysis CommentAnnotation/notes on a particular compound's data or its analysis.String
5Curve_DescriptionA description of dose-response curve quality. A complete curve has two observed asymptotes; a partial curve may not have attained its second asymptote at the highest concentration tested. High efficacy curves exhibit efficacy greater than 80% of control. Partial efficacies are statistically significant, but below 80% of control.String
6Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
7Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
8Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
9Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
10Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
11Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
12Excluded_PointsWhich dose-response titration points were excluded from analysis based on outlier analysis. Each number represents whether a titration point was (1) or was not (0) excluded, for the titration series going from smallest to highest compound concentrations.String
13Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
14Activity at 0.0000068120 uM (6.81196e-06μM**)% Activity at given concentration.Float%
15Activity at 0.0000136239 uM (1.36239e-05μM**)% Activity at given concentration.Float%
16Activity at 0.0000272478 uM (2.72478e-05μM**)% Activity at given concentration.Float%
17Activity at 0.0000544957 uM (5.44957e-05μM**)% Activity at given concentration.Float%
18Activity at 0.0001089913 uM (0.000108991μM**)% Activity at given concentration.Float%
19Activity at 0.0002179827 uM (0.000217983μM**)% Activity at given concentration.Float%
20Activity at 0.0004359654 uM (0.000435965μM**)% Activity at given concentration.Float%
21Activity at 0.0008719308 uM (0.000871931μM**)% Activity at given concentration.Float%
22Activity at 0.00174 uM (0.00174386μM**)% Activity at given concentration.Float%
23Activity at 0.00349 uM (0.00348772μM**)% Activity at given concentration.Float%
24Activity at 0.00698 uM (0.00697545μM**)% Activity at given concentration.Float%
25Activity at 0.014 uM (0.0139509μM**)% Activity at given concentration.Float%
26Activity at 0.028 uM (0.0279018μM**)% Activity at given concentration.Float%
27Activity at 0.056 uM (0.0558036μM**)% Activity at given concentration.Float%
28Activity at 0.112 uM (0.111607μM**)% Activity at given concentration.Float%
29Activity at 0.223 uM (0.223214μM**)% Activity at given concentration.Float%
30Activity at 0.446 uM (0.446429μM**)% Activity at given concentration.Float%
31Activity at 0.893 uM (0.892857μM**)% Activity at given concentration.Float%
32Activity at 1.786 uM (1.78571μM**)% Activity at given concentration.Float%
33Activity at 3.571 uM (3.57143μM**)% Activity at given concentration.Float%
34Activity at 7.143 uM (7.14286μM**)% Activity at given concentration.Float%
35Activity at 14.29 uM (14.2857μM**)% Activity at given concentration.Float%
36Activity at 28.57 uM (28.5714μM**)% Activity at given concentration.Float%
37Activity at 57.14 uM (57.1429μM**)% Activity at given concentration.Float%
38Compound QCSource of compound QCString

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: MH084681-02

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