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BioAssay: AID 2128

Late stage results from the probe development efforts to identify selective inhibitors of VIM-2 metallo-beta-lactamase: probe results

Name: Late stage results from the probe development efforts to identify selective inhibitors of VIM-2 metallo-beta-lactamase: probe results ..more
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AID: 2128
Data Source: The Scripps Research Institute Molecular Screening Center (VIM2-SELECTIVE_INH_PROBES_LATE_STAGE)
BioAssay Type: Panel
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2009-11-12
Hold-until Date: 2010-06-08
Modify Date: 2013-01-04

Data Table ( Complete ):           Active    All
BioActive Compound: 1
Depositor Specified Assays
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AIDNameTypeProbeComment
1527Primary biochemical high throughput screening assay to identify inhibitors of VIM-2 metallo-beta-lactamasescreening Primary screen (VIM-2 inhibitors).
1556Epi-absorbance primary biochemical high throughput screening assay to identify inhibitors of IMP-1 metallo-beta-lactamasescreening Primary screen (IMP-1 inhibitors).
1856Epi-absorbance-based counterscreen for selective VIM-2 inhibitors: biochemical high throughput screening assay to identify inhibitors of IMP-1 metallo-beta-lactamase.screening Counterscreen (IMP-1 inhibitors).
1857FRET-based counterscreen assay for selective VIM-2 inhibitors: biochemical high throughput screening assay to identify epi-absorbance assay artifactsscreening Counterscreen (VIM-2 inhibitors).
1860Epi-absorbance-based confirmation biochemical high throughput screening assay to identify selective inhibitors of VIM-2 metallo-beta-lactamase.screening Confirmation screen (VIM-2 inhibitors).
1866Epi-absorbance-based counterscreen assay for selective VIM-2 inhibitors: biochemical high throughput screening assay to identify inhibitors of TEM-1 serine-beta-lactamase.screening Counterscreen (TEM-1 inhibitors).
1919Epi-absorbance-based dose response biochemical high throughput screening assay for selective inhibitors of VIM-2 metallo-beta-lactamaseconfirmatory Dose response screen (VIM-2 inhibitors).
1920Epi-absorbance-based counterscreen for selective VIM-2 inhibitors: dose response biochemical high throughput screening assay to identify inhibitors of IMP-1 metallo-beta-lactamase.confirmatory Dose response counterscreen (IMP-1 inhibitors).
1925Epi-absorbance-based counterscreen for selective VIM-2 inhibitors: dose response biochemical high throughput screening assay to identify inhibitors of TEM-1 serine-beta-lactamase.confirmatory Dose response counterscreen (TEM-1 inhibitors).
1926FRET-based counterscreen for selective VIM-2 inhibitors: dose response biochemical high throughput screening assay to identify epi-absorbance assay artifacts.confirmatory Dose response counterscreen (artifacts).
1927FRET-based counterscreen for selective VIM-2 inhibitors: dose response biochemical high throughput screening assay to identify inhibitors of IMP-1 metallo-beta-lactamase.confirmatory Dose response counterscreen (IMP-1 inhibitors).
1854Summary of probe development efforts to identify selective inhibitors of VIM-2 metallo-beta-lactamasesummary1 Summary AID (VIM-2 inhibitors).
624095Late stage assay provider results from the probe development efforts to identify inhibitors of VIM-2 metallo-beta-lactamase (nonselective): Growth inhibition of clinically relevant IMP-1 transformed P. aeruginosa (KN20) in the presence of imipenem (synergy)other
624097Late stage assay provider results from the probe development efforts to identify inhibitors of VIM-2 metallo-beta-lactamase (nonselective):IMP-1-transformed E. coli growth inhibition in the presence of imipenem (synergy)other1
624081Late stage assay provider results from the probe development efforts to identify inhibitors of VIM-2 metallo-beta-lactamase (nonselective): VIM-2-transformed E. coli growth inhibition in the presence of imipenem (synergy)other1
624082Late stage assay provider results from the probe development efforts to identify inhibitors of VIM-2 metallo-beta-lactamase (nonselective): Growth inhibition of clinically relevant New Delhi metallo-beta-lactamase-1 (NDM-1)-transformed K. pneumoniae (BAA-2146) in the presence of imipenem (synergy)other
624096Late stage assay provider results from the probe development efforts to identify inhibitors of VIM-2 metallo-beta-lactamase (nonselective): Growth inhibition of clinically relevant VIM-2-transformed Acinetobacter species (YMC07/8/B3323) in the presence of imipenem (synergy)other2
504620Late stage assay provider results from the probe development efforts to identify selective inhibitors of VIM-2 metallo-beta-lactamase: VIM-2-transformed E. coli growth inhibition in the presence of imipenem (synergy)confirmatory
624080Late stage assay provider results from the probe development efforts to identify inhibitors of VIM-2 metallo-beta-lactamase (nonselective): Growth inhibition of clinically relevant VIM-2 transformed P. aeruginosa (PA641) in the presence of imipenem (synergy)other
Description:
Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center (SRIMSC)
Center Affiliation: The Scripps Research Institute (TSRI)
Assay Provider: Peter Hodder, TSRI
Network: Molecular Libraries Probe Production Centers Network (MLPCN)
Grant Proposal Number: 1 R21 NS059451-01 Fast Track
Grant Proposal PI: Peter Hodder, TSRI
External Assay ID: VIM2-SELECTIVE_INH_PROBES_LATE STAGE PROBE DATA

Name: Late stage results from the probe development efforts to identify selective inhibitors of VIM-2 metallo-beta-lactamase: probe results

Description:

The emergence of gram-negative bacteria that exhibit multi-drug resistance, combined with the paucity of new antibiotics, poses a public health challenge (1). The production of bacterial beta-lactamase enzymes, in particular, is a common mechanism of drug resistance (2-4). The beta-lactamases evolved from bacteria with resistance to naturally-occurring beta-lactams or penams (5), agents which inhibit the transpeptidase involved in cell wall biosynthesis (6). Human medicine adapted these agents into synthetic antibiotics such as penicillins, cephalosporins, carbapenems, and monobactams that contain a 2-azetidone ring (5, 7). The metallo-beta-lactamases (MBL) are zinc-dependent class B beta-lactamases that hydrolyze the beta-lactam ring, rendering the antibiotic ineffective (6, 8). Increasingly, nosocomial beta-lactam antibiotic resistance arises in P. aeruginosa, Enterobacteriaceae, and other pathogenic bacteria via gene transfer of B1 MBLs (4, 9), including IMP (active on IMiPenem) (10) and VIM (Verona IMipenemase) (11, 12). For two of these enzymes, VIM-2 and IMP-1, no inhibitors exist for clinical use (6, 9). Thus, the identification of MBL inhibitors would provide useful tools for reducing nosocomial infections and elucidating their mechanism of action (13).

Summary of Probe Development Effort:

Following nitrocefin-based primary HTS in singlicate to identify selective VIM-2 inhibitors (AID 1527), counterscreening in singlicate against the IMP-1 metallo-beta-lactamase (AID 1556), confirmation of hit activity in triplicate (AID 1860), counterscreening in triplicate against IMP-1 (AID 1856) and TEM-1 (AID 1866), VIM-2 CCF2-based counterscreening in triplicate to identify nitrocefin-based assay artifacts (AID1857), titration assays in triplicate to determine VIM-2 potency (AID 1919) and selectivity against IMP-1 (AID 1920 and 1927), TEM-1 (1925), and CCF2-based dose response counterscreening in triplicate to identify nitrocefin-based assay artifacts (AID 1926), certain compounds were identified as possible candidates for probe development. A promising lead compound (SID24790728) was order as a powder (SID 85856282) and tested in kinetic assays to determine the mode of inhibition and the inhibitory constant (Ki) against VIM2. SID 85856282 was also subsequently tested in VIM2 dose response assays, IMP1, and TEM1 counterscreens during probe development.

The results of these probe development efforts resulted in the identification of one compound (SID 85856282) as a selective, non-competitive inhibitor probe of the metallo-beta-lactamase VIM-2. This probe exhibits a submicromolar Ki value for VIM-2 with minimal activity against IMP-1 and TEM-1 enzymes, as determined using epi-absorbance and fluorescence-based biochemical assays.

Details of protocols, compound structures, and results from the original assays can be found in PubChem at the respective AIDs listed below. The results of our probe development efforts can be found at http://molscreen.florida.scripps.edu/probes.shtml. One manuscript has been published (14). Please also see Summary AID 1854.

References:

1. Siegel, R.E., Emerging gram-negative antibiotic resistance: daunting challenges, declining sensitivities, and dire consequences. Respir Care, 2008. 53(4): p. 471-9.
2. Gupta, V., An update on newer beta-lactamases. Indian J Med Res, 2007. 126(5): p. 417-27.
3. Bradford, P.A., Extended-spectrum beta-lactamases in the 21st century: characterization, epidemiology, and detection of this important resistance threat. Clin Microbiol Rev, 2001. 14(4): p. 933-51, table of contents.
4. Sacha, P., Wieczorek, P., Hauschild, T., Zorawski, M., Olszanska, D., and Tryniszewska, E., Metallo-beta-lactamases of Pseudomonas aeruginosa--a novel mechanism resistance to beta-lactam antibiotics. Folia Histochem Cytobiol, 2008. 46(2): p. 137-42.
5. Koch, A.L., Bacterial wall as target for attack: past, present, and future research. Clin Microbiol Rev, 2003. 16(4): p. 673-87.
6. Jin, W., Arakawa, Y., Yasuzawa, H., Taki, T., Hashiguchi, R., Mitsutani, K., Shoga, A., Yamaguchi, Y., Kurosaki, H., Shibata, N., Ohta, M., and Goto, M., Comparative study of the inhibition of metallo-beta-lactamases (IMP-1 and VIM-2) by thiol compounds that contain a hydrophobic group. Biol Pharm Bull, 2004. 27(6): p. 851-6.
7. Abeylath, S.C. and Turos, E., Drug delivery approaches to overcome bacterial resistance to beta-lactam antibiotics. Expert Opin Drug Deliv, 2008. 5(9): p. 931-49.
8. Wang, Z., Fast, W., Valentine, A.M., and Benkovic, S.J., Metallo-beta-lactamase: structure and mechanism. Curr Opin Chem Biol, 1999. 3(5): p. 614-22.
9. Walsh, T.R., Toleman, M.A., Poirel, L., and Nordmann, P., Metallo-beta-lactamases: the quiet before the storm? Clin Microbiol Rev, 2005. 18(2): p. 306-25.
10. Hirakata, Y., Izumikawa, K., Yamaguchi, T., Takemura, H., Tanaka, H., Yoshida, R., Matsuda, J., Nakano, M., Tomono, K., Maesaki, S., Kaku, M., Yamada, Y., Kamihira, S., and Kohno, S., Rapid detection and evaluation of clinical characteristics of emerging multiple-drug-resistant gram-negative rods carrying the metallo-beta-lactamase gene blaIMP. Antimicrob Agents Chemother, 1998. 42(8): p. 2006-11.
11. Lauretti, L., Riccio, M.L., Mazzariol, A., Cornaglia, G., Amicosante, G., Fontana, R., and Rossolini, G.M., Cloning and characterization of blaVIM, a new integron-borne metallo-beta-lactamase gene from a Pseudomonas aeruginosa clinical isolate. Antimicrob Agents Chemother, 1999. 43(7): p. 1584-90.
12. Wang, C.X. and Mi, Z.H., Imipenem-resistant Pseudomonas aeruginosa producing IMP-1 metallo-beta-lactamases and lacking the outer-membrane protein OprD. J Med Microbiol, 2006. 55(Pt 3): p. 353-4.
13. Zuck P, O'Donnell GT, Cassaday J, Chase P, Hodder P, Strulovici B, Ferrer M. Miniaturization of absorbance assays using the fluorescent properties of white microplates. Anal Biochem. 2005 Jul 15;342 (2):254-9.
14. Minond D, Saldanha SA, Subramaniam P, Spaargaren M, Spicer T, Fotsing JR, Weide T, Fokin VV, Sharpless KB, Galleni M, Bebrone C, Lassaux P, Hodder P. Inhibitors of VIM-2 by screening pharmacologically active and click-chemistry compound libraries. Bioorg Med Chem. 2009 Jul 15;17(14):5027-37.

Keywords:

Late stage, probes, powder, VIM-2, beta-lactamase, antibiotic resistance, bacteria, primary, confirmation, HTS, high throughput screen, 1536, selective, inhibitor, epi-absorbance, fluorescence, Scripps, Scripps Florida, The Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Probe Production Center Network, MLPCN.
Panel Information
Assays
    Data Table(Active)    Data Table(All)Show more
PID§NameSubstancePanel TargetsDescriptionAdditional Information
ActiveInactive
1VIM-2 Ki Assay1metallo beta-lactamase [Pseudomonas aeruginosa] [gi:7381449]
Taxonomy id: 287
2Nitrocefin-based VIM2 Inhibition Assay1metallo beta-lactamase [Pseudomonas aeruginosa] [gi:7381449]
Taxonomy id: 287
3Nitrocefin-based IMP1 Inhibition Counterscreen1metallo-beta-lactamase IMP-1 [Pseudomonas aeruginosa] [gi:27368096]
Taxonomy id: 287
4Nitrocefin-based TEM-1 Inhibition Counterscreen1Beta lactamase [Pseudomonas aeruginosa] [gi:114881106]
Taxonomy id: 287
Gene id: 4290808

§ Panel component ID.
Protocol
Please see AIDs 1527, 1556, 1856, 1857, 1860, 1866, 1919, 1920, 1925, 1926, 1927, 1854 and below for protocols performed in this probe development effort.

VIM-2 Ki Assay (Assay 1):

The purpose of this assay is to determine the inhibition constant (Ki) and modality of probe candidate molecules. Kinetic assays were conducted by incubating a range of substrate concentrations (100-8 muM) with varying inhibitor concentrations and 0.1 nM enzyme at room temperature in buffer containing 50mM HEPES, 50muM ZnSO4, 0.05% Brij 35, pH 7.1. Absorbance was measured on a Tecan Safire2 monochromatic microplate reader at 495 nm. Initial velocities were obtained from plots of absorbance at 495 nm versus time, using data points from only the linear portion of the hydrolysis curve. Substrate hydrolysis was continuously monitored. Initial velocities were plotted vs. substrate concentration and kinetic parameters were calculated using Graphpad Prism version 5.01 suite of programs. All Ki values were determined by non-linear regression (hyperbolic equation) analysis using the mixed inhibition model which allows for simultaneous determination of mechanism of inhibition. Mechanism of inhibition was determined using the "alpha" parameter derived from a mixed-model inhibition by GraphPad Prism. The mechanism of inhibition was additionally evaluated by Hanes-Woolf and Lineweaver-Burke plots. Lineweaver-Burke best-fit lines are reported, along with Ki calculated from non-linear regression. The number of replicates for this assay is 3.

Nitrocefin-based VIM2 Inhibition Assay (Assay 2):

The purpose of this assay is to identify compounds that act as inhibitors of the VIM-2 beta-lactamase. This biochemical epi-absorbance-format assay employs the cephalosporin nitrocefin as the VIM-2 substrate, and takes advantage of the fluorescent properties of white microtiter plates (13). Nitrocefin is a yellow chromogenic substrate (Imax = 395 nm) that is hydrolyzed by beta-lactamases to yield a red product with increased absorbance properties (Imax = 495 nm) that quenches plate fluorescence by absorbing the plate's emission light (13). In this assay, test compounds are incubated with purified VIM-2 enzyme and nitrocefin in detergent-containing buffer at room temperature. The reaction is stopped by the addition of EDTA, followed by measurement of well fluorescence. As designed, compounds that inhibit VIM-2 will inhibit nitrocefin hydrolysis, inhibit generation of red product, and inhibit quenching of plate fluorescence, resulting in an increase in well fluorescence. Compounds are tested in triplicate using a dilution series starting at a nominal test concentration of 60 uM.

Nitrocefin-based IMP1 Inhibition Counterscreen (Assay 3):

The purpose of this assay is to identify compounds that act as inhibitors of the IMP-1 beta-lactamase. This assay also serves as a counterscreen to identify non-selective inhibitors. This biochemical epi-absorbance-format assay employs the cephalosporin nitrocefin as the IMP-1 substrate, and takes advantage of the fluorescent properties of white microtiter plates (13). Nitrocefin is a yellow chromogenic substrate (Imax = 395 nm) that is hydrolyzed by beta-lactamases to yield a red product with increased absorbance properties (Imax = 495 nm) that quenches plate fluorescence by absorbing the plate's emission light (13). In this assay, test compounds are incubated with purified IMP-1 enzyme and nitrocefin in detergent-containing buffer at room temperature. The reaction is stopped by the addition of EDTA, followed by measurement of well fluorescence. As designed, compounds that inhibit IMP-1 will inhibit nitrocefin hydrolysis, inhibit generation of red product, and inhibit quenching of plate fluorescence, resulting in an increase in well fluorescence. Compounds are tested in triplicate using a dilution series starting at a nominal test concentration of 60 uM.

Nitrocefin-based TEM-1 Inhibition Counterscreen (Assay 4):

The purpose of this assay is to determine whether compounds identified as active in a previous set of experiments entitled, "Primary biochemical high throughput screening assay to identify inhibitors of VIM-2 metallo-beta-lactamase" (PubChem AID 1527), were nonselective or assay artifact due to inhibition of TEM-1, a serine beta-lactamase that hydrolyzes ampicillin, has negligible activity against extended-spectrum cephalosporins, and is inhibited by clavulanic acid (14). This biochemical epi-absorbance-format assay employs the cephalosporin nitrocefin as the TEM-1 substrate, and takes advantage of the fluorescent properties of white microtiter plates (13). Nitrocefin is a yellow chromogenic substrate (Imax = 395 nm) that is hydrolyzed by beta-lactamases to yield a red product with increased absorbance properties (Imax = 495 nm) that quenches plate fluorescence by absorbing the plate's emission light (13). In this assay, test compounds are incubated with purified TEM-1 enzyme and nitrocefin in detergent-containing buffer at room temperature. The reaction is stopped by the addition of potassium clavulanate, followed by measurement of well fluorescence. As designed, compounds that inhibit TEM-1 will inhibit nitrocefin hydrolysis, inhibit generation of red product, and inhibit quenching of plate fluorescence, resulting in an increase in well fluorescence. Compounds are tested in triplicate using a dilution series starting at a nominal test concentration of 60 uM.
Comment
This AID presents the results of assays that tested powder samples of a probe candidate. Probes were identified.
Result Definitions
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TIDNameDescriptionPID§Panel TargetsHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1VIM-2 Ki Assay (Ki)*The inhibition constant used to represent the binding affinity of a non-competitive inhibitor; the concentration of competing ligand in the competition assay which would occupy 50% of the receptors if no ligand was present, shown in nanomolar.1metallo beta-lactamase [Pseudomonas aeruginosa]FloatnM
2VIM-2 Ki Assay (StdErr Ki)The standard error of the Ki value.1Float
3VIM-2 Ki Assay (Slope) [0 nM] (0μM**)The slope of the Lineweaver-Burke plot, defined as the ratio Km/Vmax, in units of uM/sec at the indicated inhibitor test concentration.1Float
4VIM-2 Ki Assay (StdErr Slope) [0 nM] (0μM**)The Standard error of the slope1Float
5VIM-2 Ki Assay (Y Intercept) [0 nM] (0μM**)The y-intercept of the Lineweaver-Burke plot, defined as 1/Vmax, in units of sec-1, at the indicated inhibitor test concentration.1Float
6VIM-2 Ki Assay (StdErr Y Intercept) [0 nM] (0μM**)The Standard error of the Y intercept1Float
7VIM-2 Ki Assay (R Squared) [0 nM] (0μM**)This statistic measures how successfully the fit explains the variation of the data used to calculate the Lineweaver-Burke plot; quantifies the goodness of fit of experimental data to a model. Reported at the indicated inhibitor test concentration1Float
8VIM-2 Ki Assay (Slope) [40 nM] (0.04μM**)The slope of the Lineweaver-Burke plot, defined as the ratio Km/Vmax, in units of uM/sec at the indicated inhibitor test concentration.1Float
9VIM-2 Ki Assay (StdErr Slope) [40 nM] (0.04μM**)The Standard error of the slope1Float
10VIM-2 Ki Assay (Y Intercept) [40 nM] (0.04μM**)The y-intercept of the Lineweaver-Burke plot, defined as 1/Vmax, in units of sec-1, at the indicated inhibitor test concentration.1Float
11VIM-2 Ki Assay (StdErr Y Intercept) [40 nM] (0.04μM**)The Standard error of the Y intercept1Float
12VIM-2 Ki Assay (R Squared) [40 nM] (0.04μM**)This statistic measures how successfully the fit explains the variation of the data used to calculate the Lineweaver-Burke plot; quantifies the goodness of fit of experimental data to a model. Reported at the indicated inhibitor test concentration1Float
13VIM-2 Ki Assay (Slope) [80 nM] (0.08μM**)The slope of the Lineweaver-Burke plot, defined as the ratio Km/Vmax, in units of uM/sec at the indicated inhibitor test concentration.1Float
14VIM-2 Ki Assay (StdErr Slope) [80 nM] (0.08μM**)The Standard error of the slope1Float
15VIM-2 Ki Assay (Y Intercept) [80 nM] (0.08μM**)The y-intercept of the Lineweaver-Burke plot, defined as 1/Vmax, in units of sec-1, at the indicated inhibitor test concentration.1Float
16VIM-2 Ki Assay (StdErr Y Intercept) [80 nM] (0.08μM**)The Standard error of the Y intercept1Float
17VIM-2 Ki Assay (R Squared) [80 nM] (0.08μM**)This statistic measures how successfully the fit explains the variation of the data used to calculate the Lineweaver-Burke plot; quantifies the goodness of fit of experimental data to a model. Reported at the indicated inhibitor test concentration1Float
18VIM-2 Ki Assay (Slope) [100 nM] (0.1μM**)The slope of the Lineweaver-Burke plot, defined as the ratio Km/Vmax, in units of uM/sec at the indicated inhibitor test concentration.1Float
19VIM-2 Ki Assay (StdErr Slope) [100 nM] (0.1μM**)The Standard error of the slope1Float
20VIM-2 Ki Assay (Y Intercept) [100 nM] (0.1μM**)The y-intercept of the Lineweaver-Burke plot, defined as 1/Vmax, in units of sec-1, at the indicated inhibitor test concentration.1Float
21VIM-2 Ki Assay (StdErr Y Intercept) [100 nM] (0.1μM**)The Standard error of the Y intercept1Float
22VIM-2 Ki Assay (R Squared) [100 nM] (0.1μM**)This statistic measures how successfully the fit explains the variation of the data used to calculate the Lineweaver-Burke plot; quantifies the goodness of fit of experimental data to a model. Reported at the indicated inhibitor test concentration1Float
23VIM-2 Ki Assay (Slope) [150 nM] (0.15μM**)The slope of the Lineweaver-Burke plot, defined as the ratio Km/Vmax, in units of uM/sec at the indicated inhibitor test concentration.1Float
24VIM-2 Ki Assay (StdErr Slope) [150 nM] (0.15μM**)The Standard error of the slope1Float
25VIM-2 Ki Assay (Y Intercept) [150 nM] (0.15μM**)The y-intercept of the Lineweaver-Burke plot, defined as 1/Vmax, in units of sec-1, at the indicated inhibitor test concentration.1Float
26VIM-2 Ki Assay (StdErr Y Intercept) [150 nM] (0.15μM**)The Standard error of the Y intercept1Float
27VIM-2 Ki Assay (R Squared) [150 nM] (0.15μM**)This statistic measures how successfully the fit explains the variation of the data used to calculate the Lineweaver-Burke plot; quantifies the goodness of fit of experimental data to a model. Reported at the indicated inhibitor test concentration1Float
28VIM-2 Ki Assay (Outcome)The Assay outcome, one of Active, Inactive or Not Tested.1Outcome%
29Nitrocefin-based VIM2 Inhibition (IC50)*The concentration at which 50 percent of the activity in the inhibitor assay is observed; (IC50) shown in micromolar.2metallo beta-lactamase [Pseudomonas aeruginosa]FloatμM
30Nitrocefin-based VIM2 Inhibition (StdDev of IC50)Standard deviation of the dose response assay derived from the average IC50 of the triplicate data for the compound.2Float
31Nitrocefin-based VIM2 Inhibition (Outcome)The Assay outcome, one of Active, Inactive or Not Tested.2Outcome
32Nitrocefin-based VIM2 Inhibition (Inhibition at 3.0 nM) (0.003μM**)Value of %inhibition at 3.0 nanomolar inhibitor concentration; average of triplicate measurement.2Float%
33Nitrocefin-based VIM2 Inhibition (Inhibition at 9.1 nM) (0.0091μM**)Value of %inhibition at 9.1 nanomolar inhibitor concentration; average of triplicate measurement.2Float%
34Nitrocefin-based VIM2 Inhibition (Inhibition at 27.3 nM) (0.0273μM**)Value of %inhibition at 27.3 nanomolar inhibitor concentration; average of triplicate measurement.2Float%
35Nitrocefin-based VIM2 Inhibition (Inhibition at 81.8 nM) (0.0818μM**)Value of %inhibition at 81.8 nanomolar inhibitor concentration; average of triplicate measurement.2Float%
36Nitrocefin-based VIM2 Inhibition (Inhibition at 245.4 nM) (0.2454μM**)Value of %inhibition at 245.4 nanomolar inhibitor concentration; average of triplicate measurement.2Float%
37Nitrocefin-based VIM2 Inhibition (Inhibition at 736.3 nM) (0.7363μM**)Value of %inhibition at 736.3 nanomolar inhibitor concentration; average of triplicate measurement.2Float%
38Nitrocefin-based VIM2 Inhibition (Inhibition at 2.2 uM) (2.2μM**)Value of %inhibition at 2.2 micromolar inhibitor concentration; average of triplicate measurement.2Float%
39Nitrocefin-based VIM2 Inhibition (Inhibition at 6.6 uM) (6.6μM**)Value of %inhibition at 6.6 micromolar inhibitor concentration; average of triplicate measurement.2Float%
40Nitrocefin-based VIM2 Inhibition (Inhibition at 19.9 uM) (19.9μM**)Value of %inhibition at 19.9 micromolar inhibitor concentration; average of triplicate measurement.2Float%
41Nitrocefin-based VIM2 Inhibition (Inhibition at 59.6 uM) (59.6μM**)Value of %inhibition at 59.6 micromolar inhibitor concentration; average of triplicate measurement.2Float%
42Nitrocefin-based IMP1 Inhibition (Qualifier)Activity Qualifier identifies if the resultant data IC50 came from a fitted curve or was determined manually to be less than or greater than its listed IC50 concentration.3metallo-beta-lactamase IMP-1 [Pseudomonas aeruginosa]String
43Nitrocefin-based IMP1 Inhibition (IC50)*The concentration at which 50 percent of the activity in the inhibitor assay is observed; (IC50) shown in micromolar.3FloatμM
44Nitrocefin-based IMP1 Inhibition (Outcome)The Assay outcome, one of Active, Inactive or Not Tested.3Outcome
45Nitrocefin-based IMP1 Inhibition (Inhibition at 3.0 nM) (0.003μM**)Value of %inhibition at 3.0 nanomolar inhibitor concentration; average of triplicate measurement.3Float%
46Nitrocefin-based IMP1 Inhibition (Inhibition at 9.1 nM) (0.0091μM**)Value of %inhibition at 9.1 nanomolar inhibitor concentration; average of triplicate measurement.3Float%
47Nitrocefin-based IMP1 Inhibition (Inhibition at 27.3 nM) (0.0273μM**)Value of %inhibition at 27.3 nanomolar inhibitor concentration; average of triplicate measurement.3Float%
48Nitrocefin-based IMP1 Inhibition (Inhibition at 81.8 nM) (0.0818μM**)Value of %inhibition at 81.8 nanomolar inhibitor concentration; average of triplicate measurement.3Float%
49Nitrocefin-based IMP1 Inhibition (Inhibition at 245.4 nM) (0.2454μM**)Value of %inhibition at 245.4 nanomolar inhibitor concentration; average of triplicate measurement.3Float%
50Nitrocefin-based IMP1 Inhibition (Inhibition at 736.3 nM) (0.7363μM**)Value of %inhibition at 736.3 nanomolar inhibitor concentration; average of triplicate measurement.3Float%
51Nitrocefin-based IMP1 Inhibition (Inhibition at 2.2 uM) (2.2μM**)Value of %inhibition at 2.2 micromolar inhibitor concentration; average of triplicate measurement.3Float%
52Nitrocefin-based IMP1 Inhibition (Inhibition at 6.6 uM) (6.6μM**)Value of %inhibition at 6.6 micromolar inhibitor concentration; average of triplicate measurement.3Float%
53Nitrocefin-based IMP1 Inhibition (Inhibition at 19.9 uM) (19.9μM**)Value of %inhibition at 19.9 micromolar inhibitor concentration; average of triplicate measurement.3Float%
54Nitrocefin-based IMP1 Inhibition (Inhibition at 59.6 uM) (59.6μM**)Value of %inhibition at 59.6 micromolar inhibitor concentration; average of triplicate measurement.3Float%
55Nitrocefin-based TEM-1 Inhibition (Qualifier)Activity Qualifier identifies if the resultant data IC50 came from a fitted curve or was determined manually to be less than or greater than its listed IC50 concentration4Beta lactamase [Pseudomonas aeruginosa]String
56Nitrocefin-based TEM-1 Inhibition (IC50)*The concentration at which 50 percent of the activity in the inhibitor assay is observed; (IC50) shown in micromolar.4FloatμM
57Nitrocefin-based TEM-1 Inhibition (Outcome)The Assay outcome, one of Active, Inactive or Not Tested.4Outcome
58Nitrocefin-based TEM-1 Inhibition (Inhibition at 3.0 nM) (0.003μM**)Value of %inhibition at 3.0 nanomolar inhibitor concentration; average of triplicate measurement.4Float%
59Nitrocefin-based TEM-1 Inhibition (Inhibition at 9.1 nM) (0.0091μM**)Value of %inhibition at 9.1 nanomolar inhibitor concentration; average of triplicate measurement.4Float%
60Nitrocefin-based TEM-1 Inhibition (Inhibition at 27.3 nM) (0.0273μM**)Value of %inhibition at 27.3 nanomolar inhibitor concentration; average of triplicate measurement.4Float%
61Nitrocefin-based TEM-1 Inhibition (Inhibition at 81.8 nM) (0.0818μM**)Value of %inhibition at 81.8 nanomolar inhibitor concentration; average of triplicate measurement.4Float%
62Nitrocefin-based TEM-1 Inhibition (Inhibition at 245.4 nM) (0.2454μM**)Value of %inhibition at 245.4 nanomolar inhibitor concentration; average of triplicate measurement.4Float%
63Nitrocefin-based TEM-1 Inhibition (Inhibition at 736.3 nM) (0.7363μM**)Value of %inhibition at 736.3 nanomolar inhibitor concentration; average of triplicate measurement.4Float%
64Nitrocefin-based TEM-1 Inhibition (Inhibition at 2.2 uM) (2.2μM**)Value of %inhibition at 2.2 micromolar inhibitor concentration; average of triplicate measurement.4Float%
65Nitrocefin-based TEM-1 Inhibition (Inhibition at 6.6 uM) (6.6μM**)Value of %inhibition at 6.6 micromolar inhibitor concentration; average of triplicate measurement.4Float%
66Nitrocefin-based TEM-1 Inhibition (Inhibition at 19.9 uM) (19.9μM**)Value of %inhibition at 19.9 micromolar inhibitor concentration; average of triplicate measurement.4Float%
67Nitrocefin-based TEM-1 Inhibition (Inhibition at 59.6 uM) (59.6μM**)Value of %inhibition at 59.6 micromolar inhibitor concentration; average of triplicate measurement.4Float%
68Probe Molecule OutcomeIndicates if the compound is a probe or not. One of Probe, Active or Inactive.String

* Activity Concentration. ** Test Concentration. § Panel component ID.
Additional Information
Grant Number: 1 R21 NS059451-01

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