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BioAssay: AID 1851

Cytochrome panel assay with activity outcomes

The P450 gene superfamily is involved in the metabolism and clearance of xenobiotics. This assay used various human CYP P450 isozymes to measure the dealkylation of various pro-luciferin substrates to luciferin. The luciferin is then measured by luminescence after the addition of a luciferase detection reagent. Pro-luciferin substrate concentration in the assay was equal to its KM for its CYP P450 isozyme. Inhibitors and some substrates limit the production of luciferin, and decrease measured luminescence. ..more
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 Tested Compounds
 Tested Compounds
All(16560)
 
 
Unspecified(16560)
 
 
 Tested Substances
 Tested Substances
All(17143)
 
 
Unspecified(17143)
 
 
 Related BioAssays
 Related BioAssays
AID: 1851
Data Source: NCGC (Cytochrome panel assay)
BioAssay Type: Panel
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2009-07-08
Modify Date: 2009-07-14

Data Table ( Complete ):           View All Data
Tested Compounds:
Description:
NCGC Assay Overview:

The P450 gene superfamily is involved in the metabolism and clearance of xenobiotics. This assay used various human CYP P450 isozymes to measure the dealkylation of various pro-luciferin substrates to luciferin. The luciferin is then measured by luminescence after the addition of a luciferase detection reagent. Pro-luciferin substrate concentration in the assay was equal to its KM for its CYP P450 isozyme. Inhibitors and some substrates limit the production of luciferin, and decrease measured luminescence.
Panel Information
cytochrome p450 panel assay - cytochrome p450 panel assay
    Data Table(All)Show more
PID§NameSubstancePanel TargetsDescriptionAdditional Information
ActiveInactive
1p450-cyp2c1959137532cytochrome P450, family 2, subfamily C, polypeptide 19 [Homo sapiens] [gi:4503219]
Taxonomy id: 9606
2p450-cyp2d6277111139cytochrome P450, family 2, subfamily D, polypeptide 6 isoform 2 [Homo sapiens] [gi:68509921]
Taxonomy id: 9606
3p450-cyp3a452667751cytochrome P450, family 3, subfamily A, polypeptide 4 [Homo sapiens] [gi:13435386]
Taxonomy id: 9606
4p450-cyp1a260007256cytochrome P450, family 1, subfamily A, polypeptide 2 [Homo sapiens] [gi:73915100]
Taxonomy id: 9606
5p450-cyp2c941198782cytochrome P450, family 2, subfamily C, polypeptide 9 [Homo sapiens] [gi:13699818]
Taxonomy id: 9606

§ Panel component ID.
Protocol
NCGC Assay Protocol Summary:

1, Reagent, 2 uL, Isozyme and Pro-Luciferin
2, Library compounds, 23 nl, 40 uM to 0.24 nM dilution series
3, Controls, 23 nl, Isozyme specific inhibitor
4, Reagent, 2 uL, NADPH regeneration solution
5, Incubation time, 60 min, Isozyme specific temperature
6, Reagent, 4ul, Detection reagent
7, Incubation time, 20 min, Isozyme specific temperature
8, Detection, 1 sec, ViewLux luminescent read

The luciferase-based P450-GloTM Screening Systems were obtained from Promega (Madison, WI) for CYP 1A2 (V9770), CYP 2C9 (V9790), CYP2 C19 (V9880), CYP 2D6 (V9890), and CYP 3A4 Luciferin-PPXE (V9910) and were adapted for 1,536-well microplates and an automated protocol. The control compounds furafylline for 1A2 (F124), sulfaphenazole for CYP 2C9 (S0758), ketoconazole for CYP 2C19 (K1003), quinidine for 2D6 (Q3625), and ketoconazole for 3A4 (K1003) were purchased from Sigma Aldrich (St. Louis, MO). Recombinant P450 enzymes were obtained from baculovirus constructs expressed in insect cells (BD/Gentest).

Medium-binding white solid Kalypsys plates used. Mixtures kept on ice. 1A2: 10 nM 1A2, 100 uM Luc-ME, 100 mM KPO4; 2C9: 10 nM 2C9, 100 uM Luc-H, 25 mM KPO4; 2C19: 5 nM 2C19, 10 uM Luc-H EGE, 50 mM KPO4; 2D6: 5 nM 2D6, 30 uM Luc-ME EGE, 100 mM KPO4; 3A4: 10 nM, 25 uM Luc-PPXE. All amounts are reflected in their final concentration, working concentration is 2x.

NADPH regeneration solution: 1.3 mM NADP+, 3.3 mM glucose-6-phosphate, 3.3 mM MgCl2, 0.4 U/mL glucose-6-phosphate dehydrogenase for all isozymes, 200 mM KPO4 for 3A4. Mixture prepared at RT but kept on ice during automated run.

RT incubation in auxilliary plate hotel for 1A2, 2D6, and 3A4; 37C for 2C9 and 2C19.

Keywords: P450, CYP1A2, CYP2C19, CYP2C9, CYP2D6, CYP3A4, Luciferase, MLSMR, MLSCN, NIH Roadmap, qHTS, NCGC
Comment
Compounds with AC50 equal or less than 10 uM are considered active.
Categorized Comment - additional comments and annotations
From ChEMBL:
Assay Type: Functional
Result Definitions
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TIDNameDescriptionPID§Panel TargetsHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
1Inhibition ObservedWhether compound inhibited pro-luciferin conversion with any of the five isozymes tested.String
2Approved DrugWhether compound is active ingredient in a marketed drug.String
3CollectionOriginating collection of compound sample. Exploratory is a specialized collection of bioactive compounds. Biodiverse is a diversity-oriented collection of screening String
4Analysis CommentAnnotation/notes on a particular compound's data or its analysisString
5Activity Outcomep450-cyp2c19 Activity Outcome1cytochrome P450, family 2, subfamily C, polypeptide 19 [Homo sapiens]Outcome
6Activity Scorep450-cyp2c19 Activity Score1Integer
7p450-cyp2c19-PotencyConcentration 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.1FloatμM
8p450-cyp2c19-Curve_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 signficant, but below 80% of control.1String
9p450-cyp2c19-Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).1Float
10p450-cyp2c19-Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.1Float
11p450-cyp2c19-Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.1Float
12p450-cyp2c19-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.1Float%
13p450-cyp2c19-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.1Float%
14p450-cyp2c19-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.1Float
15p450-cyp2c19-Excluded_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.1String
16p450-cyp2c19-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).1String
17p450-cyp2c19-Activity at 0.0007 uM (0.0007μM**)% Activity at given concentration.1Float%
18p450-cyp2c19-Activity at 0.00160 uM (0.001599μM**)% Activity at given concentration.1Float%
19p450-cyp2c19-Activity at 0.00368 uM (0.003679μM**)% Activity at given concentration.1Float%
20p450-cyp2c19-Activity at 0.00820 uM (0.0082μM**)% Activity at given concentration.1Float%
21p450-cyp2c19-Activity at 0.018 uM (0.018μM**)% Activity at given concentration.1Float%
22p450-cyp2c19-Activity at 0.041 uM (0.041μM**)% Activity at given concentration.1Float%
23p450-cyp2c19-Activity at 0.091 uM (0.091μM**)% Activity at given concentration.1Float%
24p450-cyp2c19-Activity at 0.204 uM (0.204μM**)% Activity at given concentration.1Float%
25p450-cyp2c19-Activity at 0.457 uM (0.457μM**)% Activity at given concentration.1Float%
26p450-cyp2c19-Activity at 1.022 uM (1.022μM**)% Activity at given concentration.1Float%
27p450-cyp2c19-Activity at 2.286 uM (2.286μM**)% Activity at given concentration.1Float%
28p450-cyp2c19-Activity at 5.111 uM (5.111μM**)% Activity at given concentration.1Float%
29p450-cyp2c19-Activity at 11.43 uM (11.43μM**)% Activity at given concentration.1Float%
30p450-cyp2c19-Activity at 25.56 uM (25.56μM**)% Activity at given concentration.1Float%
31p450-cyp2c19-Activity at 57.14 uM (57.14μM**)% Activity at given concentration.1Float%
32Activity Outcomep450-cyp2d6 Activity Outcome2cytochrome P450, family 2, subfamily D, polypeptide 6 isoform 2 [Homo sapiens]Outcome
33Activity Scorep450-cyp2d6 Activity Score2Integer
34p450-cyp2d6-PotencyConcentration 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.2FloatμM
35p450-cyp2d6-Curve_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 signficant, but below 80% of control.2String
36p450-cyp2d6-Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).2Float
37p450-cyp2d6-Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.2Float
38p450-cyp2d6-Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.2Float
39p450-cyp2d6-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.2Float%
40p450-cyp2d6-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.2Float%
41p450-cyp2d6-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.2Float
42p450-cyp2d6-Excluded_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.2String
43p450-cyp2d6-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).2Float%
44p450-cyp2d6-Activity at 0.0007 uM (0.0007μM**)% Activity at given concentration.2Float%
45p450-cyp2d6-Activity at 0.00160 uM (0.0016μM**)% Activity at given concentration.2Float%
46p450-cyp2d6-Activity at 0.00368 uM (0.00368μM**)% Activity at given concentration.2Float%
47p450-cyp2d6-Activity at 0.00820 uM (0.0082μM**)% Activity at given concentration.2Float%
48p450-cyp2d6-Activity at 0.018 uM (0.018μM**)% Activity at given concentration.2Float%
49p450-cyp2d6-Activity at 0.041 uM% Activity at given concentration.2Float%
50p450-cyp2d6-Activity at 0.091 uM (0.091μM**)% Activity at given concentration.2Float%
51p450-cyp2d6-Activity at 0.204 uM (0.204μM**)% Activity at given concentration.2Float%
52p450-cyp2d6-Activity at 0.457 uM (0.457μM**)% Activity at given concentration.2Float%
53p450-cyp2d6-Activity at 1.022 uM (1.022μM**)% Activity at given concentration.2Float%
54p450-cyp2d6-Activity at 2.286 uM (2.286μM**)% Activity at given concentration2Float%
55p450-cyp2d6-Activity at 5.111 uM (5.111μM**)% Activity at given concentration.2Float%
56p450-cyp2d6-Activity at 11.43 uM (11.43μM**)% Activity at given concentration.2Float%
57p450-cyp2d6-Activity at 25.56 uM (25.56μM**)% Activity at given concentration.2Float%
58p450-cyp2d6-Activity at 57.14 uM (57.14μM**)% Activity at given concentration.2Float%
59Activity Outcomep450-cyp3a4 Activity Outcome3cytochrome P450, family 3, subfamily A, polypeptide 4 [Homo sapiens]Outcome
60Activity Scorep450-cyp3a4 Activity Score3Integer
61p450-cyp3a4-PotencyConcentration 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.3FloatμM
62p450-cyp3a4-Curve_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 signficant, but below 80% of control.3String
63p450-cyp3a4-Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).3Float
64p450-cyp3a4-Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.3Float
65p450-cyp3a4-Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.3Float
66p450-cyp3a4-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.3Float%
67p450-cyp3a4-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.3Float%
68p450-cyp3a4-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.3Float
69p450-cyp3a4-Excluded_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.3String
70p450-cyp3a4-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).3Float%
71p450-cyp3a4-Activity at 0.0007 uM (0.0007μM**)% Activity at given concentration.3Float%
72p450-cyp3a4-Activity at 0.00164 uM (0.00164μM**)% Activity at given concentration.3Float%
73p450-cyp3a4-Activity at 0.00366 uM (0.00366μM**)% Activity at given concentration.3Float%
74p450-cyp3a4-Activity at 0.00818 uM (0.00818μM**)% Activity at given concentration.3Float%
75p450-cyp3a4-Activity at 0.018 uM (0.018μM**)% Activity at given concentration.3Float%
76p450-cyp3a4-Activity at 0.041 uM (0.041μM**)% Activity at given concentration.3Float%
77p450-cyp3a4-Activity at 0.091 uM (0.091μM**)% Activity at given concentration.3Float%
78p450-cyp3a4-Activity at 0.204 uM (0.204μM**)% Activity at given concentration.3Float%
79p450-cyp3a4-Activity at 0.457 uM (0.457μM**)% Activity at given concentration3Float%
80p450-cyp3a4-Activity at 1.022 uM (1.022μM**)% Activity at given concentration.3Float%
81p450-cyp3a4-Activity at 2.286 uM (2.286μM**)% Activity at given concentration.3Float%
82p450-cyp3a4-Activity at 5.111 uM (5.111μM**)% Activity at given concentration.3Float%
83p450-cyp3a4-Activity at 11.43 uM (11.43μM**)% Activity at given concentration.3Float%
84p450-cyp3a4-Activity at 25.56 uM (25.56μM**)% Activity at given concentration.3Float%
85p450-cyp3a4-Activity at 57.14 uM (57.14μM**)% Activity at given concentration3Float%
86Activity Outcomep450-cyp1a2 Activity Outcome4cytochrome P450, family 1, subfamily A, polypeptide 2 [Homo sapiens]Outcome
87Activity Scorep450-cyp1a2 Activity Score4Integer
88p450-cyp1a2-PotencyConcentration 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.4FloatμM
89p450-cyp1a2-Curve_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 signficant, but below 80% of control.4String
90p450-cyp1a2-Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).4Float
91p450-cyp1a2-Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.4Float
92p450-cyp1a2-Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.4Float
93p450-cyp1a2-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.4Float%
94p450-cyp1a2-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.4Float%
95p450-cyp1a2-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.4Float
96p450-cyp1a2-Excluded_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.4String
97p450-cyp1a2-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).4Float
98p450-cyp1a2-Activity at 0.0007 uM (0.0007μM**)% Activity at given concentration.4Float%
99p450-cyp1a2-Activity at 0.00160 uM (0.0016μM**)% Activity at given concentration.4Float%
100p450-cyp1a2-Activity at 0.00368 uM (0.00368μM**)% Activity at given concentration.4Float%
101p450-cyp1a2-Activity at 0.00820 uM (0.0082μM**)% Activity at given concentration.4Float%
102p450-cyp1a2-Activity at 0.018 uM (0.018μM**)% Activity at given concentration.4Float%
103p450-cyp1a2-Activity at 0.041 uM (0.041μM**)% Activity at given concentration.4Float%
104p450-cyp1a2-Activity at 0.091 uM (0.091μM**)% Activity at given concentration.4Float%
105p450-cyp1a2-Activity at 0.204 uM (0.204μM**)% Activity at given concentration.4Float%
106p450-cyp1a2-Activity at 0.457 uM (0.457μM**)% Activity at given concentration.4Float%
107p450-cyp1a2-Activity at 1.022 uM (1.022μM**)% Activity at given concentration.4Float%
108p450-cyp1a2-Activity at 2.286 uM (2.286μM**)% Activity at given concentration.4Float%
109p450-cyp1a2-Activity at 5.111 uM (5.111μM**)% Activity at given concentration.4Float%
110p450-cyp1a2-Activity at 11.43 uM (11.43μM**)% Activity at given concentration.4Float%
111p450-cyp1a2-Activity at 25.56 uM (25.56μM**)% Activity at given concentration.4Float%
112p450-cyp1a2-Activity at 57.14 uM (57.14μM**)% Activity at given concentration.4Float%
113Activity Outcomep450-cyp2c9 Activity Outcome5cytochrome P450, family 2, subfamily C, polypeptide 9 [Homo sapiens]Outcome
114Activity Scorep450-cyp2c9 Activity Score5Integer
115p450-cyp2c9-PotencyConcentration 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.5FloatμM
116p450-cyp2c9-Curve_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 signficant, but below 80% of control.5String
117p450-cyp2c9-Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).5Float
118p450-cyp2c9-Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.5Float
119p450-cyp2c9-Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.5Float
120p450-cyp2c9-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.5Float%
121p450-cyp2c9-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.5Float%
122p450-cyp2c9-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.5Float
123p450-cyp2c9-Excluded_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.5String
124p450-cyp2c9-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).5Float
125p450-cyp2c9-Activity at 0.0007 uM (0.0007μM**)% Activity at given concentration.5Float%
126p450-cyp2c9-Activity at 0.00160 uM (0.0016μM**)% Activity at given concentration.5Float%
127p450-cyp2c9-Activity at 0.00368 uM (0.00368μM**)% Activity at given concentration.5Float%
128p450-cyp2c9-Activity at 0.00820 uM (0.0082μM**)% Activity at given concentration.5Float%
129p450-cyp2c9-Activity at 0.018 uM (0.018μM**)% Activity at given concentration.5Float%
130p450-cyp2c9-Activity at 0.041 uM (0.041μM**)% Activity at given concentration.5Float%
131p450-cyp2c9-Activity at 0.091 uM (0.091μM**)% Activity at given concentration.5Float%
132p450-cyp2c9-Activity at 0.204 uM (0.204μM**)% Activity at given concentration.5Float%
133p450-cyp2c9-Activity at 0.457 uM (0.457μM**)% Activity at given concentration.5Float%
134p450-cyp2c9-Activity at 1.022 uM (1.022μM**)% Activity at given concentration.5Float%
135p450-cyp2c9-Activity at 2.286 uM (2.286μM**)% Activity at given concentration.5Float%
136p450-cyp2c9-Activity at 5.111 uM (5.111μM**)% Activity at given concentration.5Float%
137p450-cyp2c9-Activity at 11.43 uM (11.43μM**)% Activity at given concentration.5Float%
138p450-cyp2c9-Activity at 25.56 uM (25.56μM**)% Activity at given concentration.5Float%
139p450-cyp2c9-Activity at 57.14 uM (57.14μM**)% Activity at given concentration.5Float%
140Compound QCString

** Test Concentration. § Panel component ID.

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