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BioAssay: AID 652076

Late-stage results from the probe development effort to identify antagonists of OPRK1: LCMS-based biochemical cytochrome P450 inhibition assay

Name: Late-stage results from the probe development effort to identify antagonists of OPRK1: LCMS-based biochemical cytochrome P450 inhibition assay. ..more
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 Tested Compounds
 Tested Compounds
All(1)
 
 
Unspecified(1)
 
 
 Tested Substances
 Tested Substances
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Unspecified(1)
 
 
 Related BioAssays
 Related BioAssays
AID: 652076
Data Source: The Scripps Research Institute Molecular Screening Center (CYTP450_INH_LCMS_OPRK1_PK)
BioAssay Type: Panel
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2013-03-05
Hold-until Date: 2013-10-21
Modify Date: 2013-10-21

Data Table ( Complete ):           All
Tested Compound:
Depositor Specified Assays
AIDNameTypeComment
652031Maybridge screen to identify antagonists of kappa opioid receptor 1 (OPRK1): fluorescence-based cell-based assayscreeningMaybridge screen (OPRK1 inhibitors in singlicate)
652032Late-stage results from the probe development effort to identify antagonists of OPRK1: fluorescence-based cell-based dose response assayconfirmatoryLate-stage dose response (OPRK1 antagonists in quadruplicate)
652033Late-stage results from the probe development effort to identify antagonists of OPRK1: fluorescence-based cell-based dose response OPRD1 counterscreenconfirmatoryLate-stage dose response counterscreen (OPRD1 antagonists in quadruplicate)
652034Late-stage results from the probe development effort to identify antagonists of OPRK1: luminescence-based cell-based dose response OPRM1 counterscreenconfirmatoryLate-stage dose response counterscreen (OPRM1 antagonists in quadruplicate)
652045Summary of the probe development efforts to identify antagonists of the kappa 1 (OPRK1) opioid receptorsummarySummary (OPRK1 antagonists)
652108Late-stage results from the probe development effort to identify antagonists of OPRK1: In vivo tail flick assayother
Description:
Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center (SRIMSC)
Affiliation: The Scripps Research Institute, TSRI
Assay Provider: Lakshmi A. Devi, Mount Sinai School of Medicine
Network: Molecular Library Probe Production Centers Network (MLPCN)
Grant Proposal Number: R03NS053751
Grant Proposal PI: Lakshmi A. Devi, Mount Sinai School of Medicine
External Assay ID: CYTP450_INH_LCMS_OPRK1_PK

Name: Late-stage results from the probe development effort to identify antagonists of OPRK1: LCMS-based biochemical cytochrome P450 inhibition assay.

Description:

Potent and selective OPRK antagonists will be useful for studying the mechanisms involved in OPRK-mediated analgesia and may have therapeutic value as novel analgesics with an improved side effect profile to currently available drugs. Studies have identified a role for dynorphin and OPRK stimulation in neuropathic pain (1). The dynorphins act as endogenous agonists at the opioid receptors, including OPRK (2), and the increased dynorphin expression in neuropathic pain also leads to a sustained activation of OPRK (1, 3). The mechanisms and neural circuits in OPRK-mediated analgesia are active areas of study; it is hoped those studies will assist in the development of novel analgesics that bypass OPRK-mediated depression (4-5). A role for dynorphin/OPRK in modulating drug addiction has been proposed (for review, see (6-7)). The function of dynorphin/OPRK systems in addiction appears to be diverse, and may modulate drug-seeking behavior depending on factors such as drug history, pattern of intake, and stress (for review, see (6)). The availability of potent and selective OPRK ligands may help unravel these mechanisms, as well as prove to be of therapeutic utility. Evidence from preclinical studies indicates that the dynorphin/OPRK system may be dysregulated in affective psychiatric disorders (for review, see (6, 8)). However, solid evidence from clinical studies is lacking. There is increasing evidence for a potential involvement of dynorphin/OPRK in schizophrenia; OPRK agonists appear to induce symptoms in humans and animals that are present in schizophrenia (8-10). Thus, the availability of new research tools such as potent and selective OPRK antagonists will facilitate understanding the physiological and pathophysiological mechanisms of dynorphin/OPRK systems and their roles in psychiatric disease in humans.

References:

1. Xu, M., et al., Neuropathic pain activates the endogenous kappa opioid system in mouse spinal cord and induces opioid receptor tolerance. J Neurosci, 2004. 24(19): p. 4576-84.
2. Chavkin, C., I.F. James, and A. Goldstein, Dynorphin is a specific endogenous ligand of the kappa opioid receptor. Science, 1982. 215(4531): p. 413-5.
3. Xu, M., et al., Sciatic nerve ligation-induced proliferation of spinal cord astrocytes is mediated by kappa opioid activation of p38 mitogen-activated protein kinase. J Neurosci, 2007. 27(10): p. 2570-81.
4. Al-Hasani, R. and M.R. Bruchas, Molecular mechanisms of opioid receptor-dependent signaling and behavior. Anesthesiology, 2011. 115(6): p. 1363-81.
5. Muschamp, J.W., A. Van't Veer, and W.A. Carlezon, Jr., Tracking down the molecular substrates of stress: new roles for p38alpha MAPK and kappa-opioid receptors. Neuron, 2011. 71(3): p. 383-5.
6. Tejeda, H.A., T.S. Shippenberg, and R. Henriksson, The dynorphin/kappa-opioid receptor system and its role in psychiatric disorders. Cell Mol Life Sci, 2012. 69(6): p. 857-96.
7. Yoo, J.H., I. Kitchen, and A. Bailey, The endogenous opioid system in cocaine addiction: what lessons have opioid peptide and receptor knockout mice taught us? Br J Pharmacol, 2012. 166(7): p. 1993-2014.
8. Schwarzer, C., 30 years of dynorphins--new insights on their functions in neuropsychiatric diseases. Pharmacol Ther, 2009. 123(3): p. 353-70.
9. Bortolato, M. and M.V. Solbrig, The price of seizure control: dynorphins in interictal and postictal psychosis. Psychiatry Res, 2007. 151(1-2): p. 139-43.
10. Sheffler, D.J. and B.L. Roth, Salvinorin A: the "magic mint" hallucinogen finds a molecular target in the kappa opioid receptor. Trends Pharmacol Sci, 2003. 24(3): p. 107-9.

Keywords:

OPRK1, kappa, opioid, receptor, mouse, pharmacokinetics, PK assay, human cytochrome P450, CYP1A2, CYP2C9, CYP2D6, CYP3A4, LCMS, LC-MS/MS, antagonist, inhibitor, inhibit, pain, analgesic, neuropathic pain, drug addiction, addiction, Scripps, Scripps DMPK Laboratory, The Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Probe Production Centers Network, MLPCN.
Panel Information
Assays
    Data Table(All)Show more
PID§NameSubstancePanel TargetsDescriptionAdditional Information
ActiveInactive
1CYP1A21cytochrome P450 1A2 [Homo sapiens] [gi:73915100]
Taxonomy id: 9606
Gene id: 1544
2CYP2C91Cytochrome P450, family 2, subfamily C, polypeptide 9 [Homo sapiens] [gi:115529027]
Taxonomy id: 9606
Gene id: 1559
3CYP2D61Cytochrome P450, family 2, subfamily D, polypeptide 6 [Homo sapiens] [gi:117558701]
Taxonomy id: 9606
Gene id: 1565
4CYP3A41cytochrome P450, family 3, subfamily A, polypeptide 4 [Homo sapiens] [gi:51094614]
Taxonomy id: 9606
Gene id: 1576

§ Panel component ID.
Protocol
Assay Overview:

The purpose of this assay is to obtain information regarding potential drug-drug interactions for a lead OPRK antagonist test compound. Inhibition of the four major human isoforms of cytochrome P450 are evaluated by following the metabolism of specific marker substrates CYP1A2, CYP2C9, CYP2D6, and CYP3A4 in the presence test compound.

Protocol Summary:

The metabolism of CYP1A2 (phenaceten demethylated to acetaminophen), CYP2C9 (tolbutamide hydroxylated to hydroxytolbutamide), CYP2D6 (bufuralol hydroxylated to 4'-hydroxybufuralol), and CYP3A4 (midazolam hydroxylated to 1'-hydroxymidazolam) in the presence or absence of 10 uM test compound are evaluated. The concentration of each marker substrate is approximately its Km. Specific inhibitors for each isoform are used to validate the system. Compound concentrations are determined by LC-MS/MS.

PubChem Activity Outcome and Score:

The following applies to each panel in this assay:

Compounds that exhibit less than or equal to 50% inhibition are considered active. Compounds that exhibit greater than 50% inhibition are considered inactive.

List of Reagents:

Reagents were provided by the DMPK Laboratory at The Scripps Research Institute in Florida.
Comment
This assay was performed by the DMPK Laboratory at The Scripps Research Institute in Florida with powder samples of synthesized compound.
Result Definitions
TIDNameDescriptionPID§Panel TargetsHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
1Outcome [CYP1A2]Outcome of assay, one of active or inactive1cytochrome P450 1A2 [Homo sapiens]Outcome
2Inhibition at 10 uM [CYP1A2] (10μM**)The percentage inhibition of the metabolism of human cytochrome 1A2 in the presence of 10 uM test compound.1Float%
3Outcome [CYP2C9]Outcome of assay, one of active or inactive2Cytochrome P450, family 2, subfamily C, polypeptide 9 [Homo sapiens]Outcome
4Inhibition at 10 uM [CYP2C9] (10μM**)The percentage inhibition of the metabolism of human cytochrome 2C9 in the presence of 10 uM test compound.2Float%
5Outcome [CYP2D6]Outcome of assay, one of active or inactive3Cytochrome P450, family 2, subfamily D, polypeptide 6 [Homo sapiens]Outcome
6Inhibition at 10 uM [CYP2D6] (10μM**)The percentage inhibition of the metabolism of human cytochrome 2D6 in the presence of 10 uM test compound.3Float%
7Outcome [CYP3A4]Outcome of assay, one of active or inactive4cytochrome P450, family 3, subfamily A, polypeptide 4 [Homo sapiens]Outcome
8Inhibition at 10 uM [CYP3A4] (10μM**)The percentage inhibition of the metabolism of human cytochrome 3A4 in the presence of 10 uM test compound.4Float%

** Test Concentration. § Panel component ID.
Additional Information
Grant Number: R03NS053751

Classification
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