DNA encoding a human serotonin receptor (5-HT4B) and uses...

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...

Reexamination Certificate

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C435S325000, C435S356000, C435S357000, C435S358000, C435S365000

Reexamination Certificate

active

06300087

ABSTRACT:

BACKGROUND OF THE INVENTION
Throughout this application various publications are referred to by partial citations within parenthesis. Full citations for these publications may be found at the end of the specification immediately preceding the claims. The disclosures of these publications, in their entireties, are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains.
The subject invention of this application, DNA encoding a human serotonin receptor (5-HT
4B
) and uses thereof, refers to the “5-HT
4B
receptor” which has been renamed to the “5-HT
7
receptor” by the “Serotonin Receptor Nomenclature Committee” of the IUPHAR. Thus, all claims herein directly or indirectly related to the 5-HT
4B
receptor and nucleic acid molecules encoding the 5-HT
4B
receptor are drawn to the 5-HT
7
receptor and nucleic acid molecules encoding the 5-HT
7
receptor also.
Primary amino acid sequence and signal transduction data are now published for four cloned 5-HT
1
-like receptors, three cloned 5-HT
2
receptors, and one 5-HT
3
receptor. Analysis of the sequence homology as well as the signal transduction pathways of these receptors leads to their grouping on the basis of these attributes: The 5-HT
1
subfamily includes: 5-HT
1A
(Fargin, 1988; Kobilka, 1989), 5-HT
1B
/5-HT
1D&bgr;
(Weinshank et al., 1991 and the rest), 5-HT
1D&agr;
, (Branchek et al. 1991; Hamblin and Metcalf, 1991; Weinshank et al., 1992), 5-HT
1E
(Levy et al., 1992; McAllister et al., 1992, Zgombick et al., 1992) and 5-HT
1F
(Adham et al, 1993). These subtypes share >50% transmembrane amino acid identity and couple to the inhibition of adenylate cyclase. The 5-HT
2
family includes the 5-HT
2
receptor (Pritchett et al., 1988), 5-HT
1C
(Julius et al., 1989) and 5-HT
2F
(Rat Stomach Fundus; Foquet et al., 1992; Kursar et al., 1992). These receptors share >70% amino acid identity and coupling to phosphoinositide hydrolysis. The 5-HT
3
receptor has been shown to be a ligand-gated ion channel (Maricq et al., 1991). Heterogeneity of 5-HT
3
receptors is controversial, although other ligand-gated ion channels display significant heterogeneity. Notably absent from this series are the 5-HT
4
receptors. The second messenger coupling from tissue studies indicates activation of adenylate cyclase as a primary mode of signal transduction (Dumius et al., 1988; Bockaert et al., 1990). We report here the cloning of the first mammalian 5-HT receptor that couples to the stimulation of adenylate cyclase activity which we propose to name 5-HT
4B
. The pharmacological properties of this receptor indicate that it may be similar to a series of functionally defined 5-HT receptors described in the porcine vena cava (Trevethick et al., 1984), cat saphenous vein, coronary arteries [Cushing and Cohen, 1992, and several vascular dilatory effects (Mylecharane and Phillips, 1989). These receptors appear to underlie contractile and relaxant responses in isolated blood vessels indicating potential therapeutic benefit in angina, coronary artery disease, atherosclerosis, and possibly cerebral blood vessel disorders leading to stroke. The presence of this subtype in the CNS also indicates potential use in disorders of higher cognitive processes as well as control of autonomic function.
SUMMARY OF THE INVENTION
This invention provides an isolated nucleic acid molecule encoding a mammalian 5-HT
4B
receptor. In a preferred embodiment of this invention, the isolated nucleic acid encodes a human 5-HT
4B
receptor. In another embodiment of this invention, the nucleic acid molecule comprises a plasmid designated pcEXV-5-HT
4B
(ATCC Accession No. 75332).
This invention provides a nucleic acid probe comprising a nucleic acid molecule of at least 15 nucleotides capable of specifically hybridizing with a sequence included within the sequence of a nucleic acid molecule encoding a mammalian 5-HT
4B
receptor. This invention also provides a nucleic acid molecule of at least 15 nucleotides capable of specifically hybridizing with a sequence included within the sequence of a nucleic acid molecule encoding a human 5-HT
4B
receptor.
This invention provides an antisense oligonucleotide having a sequence capable of binding specifically to an mRNA molecule encoding a mammalian 5-HT
4B
receptor so as to prevent translation of the mRNA molecule. This invention also provides an antisense oligonucleotide having a sequence capable of binding specifically to an mRNA molecule encoding a human 5-HT
4B
receptor so as to prevent translation of the mRNA molecule.
This invention provides a monoclonal antibody directed to a mammalian 5-HT
4B
receptor. This invention also provides a monoclonal antibody directed to a human 5-HT4B receptor.
This invention provides a pharmaceutical composition comprising an amount of a substance effective to alleviate the abnormalities resulting from overexpression of a mammalian 5-HT
4B
receptor and a pharmaceutically acceptable carrier as well as a pharmaceutical composition comprising an amount of a substance effective to alleviate abnormalities resulting from underexpression of mammalian 5-HT
4B
receptor and a pharmaceutically acceptable carrier.
This invention provides a pharmaceutical composition comprising an amount of a substance effective to alleviate the abnormalities resulting from overexpression of a human 5-HT
4B
receptor and a pharmaceutically acceptable carrier. This invention also provides pharmaceutical composition comprising an amount of a substance effective to alleviate abnormalities resulting from underexpression of a human 5-HT
4B
receptor and a pharmaceutically acceptable carrier.
This invention provides a transgenic, nonhuman mammal whose genome comprises DNA encoding a mammalian 5-HT
4B
receptor so positioned within such genome as to be transcribed into antisense mRNA complementary to mRNA encoding the mammalian 5-HT
4B
receptor and when hybridized to mRNA encoding the mammalian 5-HT
4B
receptor, the complementary mRNA reduces the translation of the mRNA encoding the mammalian 5-HT
4B
receptor.
This invention also provides a transgenic, nonhuman mammal whose genome comprises DNA encoding a human 5-HT
4B
so positioned within such genome as to be transcribed into antisense mRNA complementary to mRNA encoding the human 5-HT
4B
and when hybridized to mRNA encoding the human 5-HT4B, the complementary mRNA reduces the translation of the mRNA encoding the human 5-HT
4B
.
This invention provides a transgenic, nonhuman mammal whose genome comprises DNA encoding a mammalian 5-HT
4B
receptor positioned within such genome as to be transcribed into antisense mRNA which is complementary to mRNA encoding the mammalian 5-HT
4B
receptor and when hybridized to mRNA encoding the 5-HT
4B
receptor, the antisense mRNA thereby prevents the translation of mRNA encoding the 5-HT
4B
receptor.
This invention also provides a transgenic, nonhuman mammal whose genome comprises DNA encoding a human 5-HT4B receptor positioned within such genome as to be transcribed into antisense mRNA which is complementary to mRNA encoding the human 5-HT
4B
receptor and when hybridized to mRNA encoding human 5-HT
4B
receptor, the antisense mRNA thereby prevents the translation of mRNA encoding the 5-HT
4B
receptor.
This invention provides a method of screening drugs to identify drugs which specifically interact with, and bind to, a mammalian
5
-HT
4B
receptor on the surface of a cell which comprises contacting a mammalian cell comprising an isolated DNA molecule encoding a mammalian 5-HT
4B
receptor, the protein encoded thereby is expressed on the cell surface, with a plurality of drugs, determining those drugs which bind to the mammalian cell, and thereby identifying drugs which specifically interact with, and bind to, a mammalian 5-HT
4B
receptor.
This invention provides a method of screening drugs to identify drugs which specifically interact with, and bind to, a human 5-HT
4B
receptor on the surface of a cell which comprises contacting a mammalian c

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