Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Reexamination Certificate
1994-01-28
2001-07-10
Kunz, Gary L. (Department: 1647)
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
C530S350000, C930S010000, C536S023500, C536S024310, C435S242000, C435S320100
Reexamination Certificate
active
06258556
ABSTRACT:
FIELD OF THE INVENTION
The present application relates to isolated cDNA and genomic clones encoding human &mgr; opiate receptors. The application further relates to a purified protein having the biochemical and pharmacologic characteristics of a human &mgr; opiate receptor, cell lines which express such a protein by virtue of being transformed with DNA encoding a human &mgr; opiate receptor and to methods for determining the allelic composition of a human genome with respect to the &mgr; opiate receptor locus.
BACKGROUND OF THE INVENTION
Opiate receptors [1-5], sites recognizing exogenous opiate drugs and endogenous opiate peptides, include the morphine-preferring &mgr; opiate receptor first defined by Martin and colleagues [2]. &mgr; receptor distributions and pharmacologic properties place them among the receptors most identified with the analgesic and addicting properties of opiate drugs [3-7]. These receptors are G-linked members of the seven transmembrane domain neuropeptide receptor subfamily [8-14].
Recent studies have identified the cDNAs encoding rodent &mgr; [15-17], &dgr; [17-19] and &kgr; opiate receptors [20-22], thus defining at least one member of each of the other major opiate receptor subclasses postulated by Martin, Kosterlitz, Hughes, and associates [1-5]. The rat &mgr; opiate receptor (&mgr; receptor) has the structure of a G-protein coupled receptor. G-protein receptor coupling was confirmed for the rat &mgr; receptor [15-17]; morphine affects adenyl cyclase levels in cells expressing &mgr; receptor [5,23].
Because of interest in &mgr; receptors as targets for development of selective analgesic and anti-addictive therapies [24-28], and because of interest in identifying &mgr; receptor gene markers that could detect individuals possessing allelic variants of this gene that could confer differential susceptibility to abused drugs, we have used the rat &mgr; opiate receptor cDNA identified in this laboratory [15] to identify its human homolog.
SUMMARY OF THE INVENTION
The present invention resides, in part, in a molecular clone of DNA encoding a human &mgr; opiate receptor. Herein is described the nucleotide sequence of a cDNA encoding a human &mgr; opiate receptor. Also, the sodium- and GTP analog-sensitive high-affinity binding that its expression confers on COS cells is described. Furthermore, the changes in adenyl cyclase in expressing COS cells that are induced by treatment with opiate drugs are shown. A human &mgr; opiate receptor locus (h&mgr;OR1) is assigned to a human chromosomal region using the cloned cDNA as a probe.
The invention further resides in DNA clones encompassing a portion of the locus including the coding region of the human &mgr; opiate receptor protein. Overlapping, contiguous clones of DNA from the h&mgr;OR1 locus are described (HG3, HG4, HG24 and HG31), as is a polymorphic genetic marker at the human &mgr; opiate receptor locus.
Data presented herein document the biochemical and genetic nature of the principal human receptor for analgesic and addicting opiate ligands.
Accordingly, it is a first object of the present invention to provide cloned DNA molecules which encode a human &mgr; opiate receptor. These cloned molecules can be used as probes in assays that examine the structure and function of human &mgr; opiate receptor genes. The cloned DNA can also be used to transform a host cell so as to create cells which express human &mgr; opiate receptor on their surface.
Thus, an additional object of the present invention is to provide assays for the structure and function of the &mgr; opiate receptor in a human patient. Another object of the present invention is to provide host cells transformed with cloned DNA encoding a human &mgr; opiate receptor protein, so as to obtain expression of the cloned DNA in the host cell.
Yet another object of the invention is to provide a purified protein having the biochemical properties of a human &mgr; opiate receptor, especially wherein such protein has the amino acid sequence of SEQ ID NO:2.
The biochemical actions of human &mgr; opiate receptor upon ligand binding represent early steps in the analgesic and behavioral effects of opiate and opioid peptide ligands. Accordingly, host cells which express human &mgr; opiate receptor on their surface can be used to assay compounds for activity as agonists, mimics or antagonists of opiate and opioid peptide ligands. Thus, another object of the present invention is to provide assays for screening compounds for opiate (or enkephalin) agonist or antagonist activity.
Again, since the &mgr; receptor is the major receptor mediating opiate pharmacologic and behavioral effects, the structure of the &mgr; receptor gene in a subject is expected to have some influence upon the susceptibility of the subject to the analgesic and addictive effects of opiate compounds. Thus, yet another object of the present invention is to provide a means of evaluating the genetic susceptibility of a subject to opiate analgesia and opiate drug abuse.
REFERENCES:
patent: WO9319086 (1993-09-01), None
patent: WO 9507983 (1995-03-01), None
LeConiat et al, “Human interferon gamma receptor 1(IFNGR1) gene maps to chromosome region 6q23-6q24”,Hum. Genet.84:92-94 (Dec. 1989).*
Laureys et al, “Chromosomal Mapping of the Gene for the Type II Insulin-like Growth Factor Receptor . . . ”, Genomics 3(3):224-229 (Oct. 1988).*
Wallace et al, Methods in Enzymology, 152, 432-442, 1987.*
Ueda et al. (1988) Proc. Natl. Acad. Sci, USA, 85:7013-7017.
Yinchang et al. (1989) Proc. CAMS and PUMC, V.4, No. 1:1-7.
Xie et al. (1992) Proc. Natl. Acad. Sci. USA, 89:4124-4128.
Libert et al. (1989) Science, vol. 244:569-574.
Evans et al. (1992) Science, V.258:1952-1955.
Chen et al. (1993)Molecular Pharmacology44:8-12.
Kieffer et al. (1992)Proc Natl Acad Sci USA89:12048-12052.
Evans et al. (1992)Science258:1952-1955.
Fukuda et al. (1993)FEBS Letters327 (3) :311-314.
Nishi et al. (1993)FEBS Letters330 (1) :77-80.
Wang et al. (1993)Proc Natl Acad Sci USA90:10230-10234.
Fukuda et al. (1994)FEB Letters343:42-46.
Yasuda et al. (1994)Proc Natl Acad Sci USA90:6736-6740.
Minami et al. (1993)FEBS Letters329(3):291-295.
Johnson Peter
Persico Antonio M.
Uhl George
Wang Jia Bei
Fitch Even Tabin & Flannery
Kunz Gary L.
Landsman Robert S.
The United States of America as represented by the Department of
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