title: Inhibition of morphine-induced cAMP overshoot: a cell-based assay model in a high-throughput format.
abstract: Opiates are not only potent analgesics but also drugs of abuse mainly because they produce euphoria. Chronic use of opiates results in the development of tolerance and dependence. Dr Marshall Nirenberg's group at the National Institutes of Health (NIH) was the first to use a cellular model system of Neuroblastoma × Glioma hybrid cells (NG108-15) to study morphine addiction. They showed that opiates affect adenylyl cyclase (AC) by two opposing mechanisms mediated by the opiate receptor. Although the cellular mechanisms that cause addiction are not yet completely understood, the most observed correlative biochemical adaptation is the upregulation of AC. This model also provides the opportunity to look for compounds which could dissociate the acute effect of opiates from the delayed response, upregulation of AC, and thus lead to the discovery of non-addictive drugs. To identify small molecule compounds that can inhibit morphine-induced cAMP overshoot, we have validated and optimized a cell-based assay in a high throughput format that measures cellular cAMP production after morphine withdrawal. The assay performed well in the 1536-well plate format. The LOPAC library of 1,280 compounds was screened in this assay on a quantitative high-throughput screening (qHTS) platform. A group of compounds that can inhibit morphine-induced cAMP overshoot were identified. The most potent compounds are eight naloxone-related compounds, including levallorphan tartrate, naloxonazine dihydrochloride, naloxone hydrochloride, naltrexone hydrochloride, and naltriben methanesulfonate. The qHTS approach we used in this study will be useful in identifying novel inhibitors of morphine induced addiction from a larger scale screening.
authors: NIH Chemical Genomics Center, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA. mxia@mail.nih.gov
authors: Menghang
authors: M
authors: Xia
authors: Vicky
authors: V
authors: Guo
authors: Ruili
authors: R
authors: Huang
authors: Sampada A
authors: SA
authors: Shahane
authors: Christopher P
authors: CP
authors: Austin
authors: Marshall
authors: M
authors: Nirenberg
authors: Shail K
authors: SK
authors: Sharma
references: Proc Natl Acad Sci U S A. 1974 Sep;71(9):3474-7
references: 4530316
references: Biochem Biophys Res Commun. 2010 Feb 19;392(4):603-7
references: 20100459
references: Science. 1997 Oct 3;278(5335):58-63
references: 9311927
references: J Pharmacol Exp Ther. 2002 Jul;302(1):1-7
references: 12065693
references: Biol Psychiatry. 2008 Jun 1;63(11):1013-21
references: 18222416
references: Fed Proc. 1967 Jul-Aug;26(4):1115-8
references: 6028774
references: Biochem J. 1993 Oct 1;295 ( Pt 1):263-71
references: 8216227
references: J Biol Chem. 1995 Dec 15;270(50):29732-8
references: 8530363
references: Mol Pharmacol. 2006 Jun;69(6):1810-9
references: 16525132
references: Proc Natl Acad Sci U S A. 1977 Aug;74(8):3365-9
references: 269396
references: Curr Chem Genomics. 2009 Jan 1;3(1):7-12
references: 19936037
references: Mol Pharmacol. 2009 Sep;76(3):526-33
references: 19549762
references: Life Sci. 2004 Dec 17;76(5):479-85
references: 15556161
references: Annu Rev Pharmacol Toxicol. 1990;30:123-47
references: 2160790
references: Am J Psychiatry. 2005 Aug;162(8):1423-31
references: 16055763
references: Proc Natl Acad Sci U S A. 1975 Aug;72(8):3092-6
references: 1059094
references: J Pharmacol Exp Ther. 1999 May;289(2):607-24
references: 10215631
references: Science. 1988 Nov 4;242(4879):715-23
references: 2903550
references: J Biol Chem. 1999 Apr 2;274(14):9207-15
references: 10092593
references: Pharmacol Biochem Behav. 1981 Nov;15(5):677-80
references: 7312906
references: Proc Natl Acad Sci U S A. 1975 Feb;72(2):590-4
references: 1054841
references: J Pharmacol Exp Ther. 1997 Jun;281(3):1350-6
references: 9190871
references: J Neurochem. 1994 Nov;63(5):1726-30
references: 7931327
references: J Neurochem. 1990 Oct;55(4):1390-6
references: 1697894
references: Annu Rev Pharmacol Toxicol. 2000;40:389-430
references: 10836142
references: Nature. 1974 Mar 1;248(5443):24-7
references: 4361995
references: Proc Natl Acad Sci U S A. 2009 Feb 17;106(7):2412-7
references: 19196967