Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Patent
1995-04-13
1998-05-12
Teng, Sally P.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
4352523, 43525411, 4353201, 435325, 536 235, 530350, 530395, C12N 1509, C07K 14705
Patent
active
057503699
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
1. Technical Field
The present invention relates, in general, to the molecular cloning and expression of a receptor protein, and, in particular, to a prostaglandin F2.alpha. receptor and fragments thereof linked to the activation of second messengers as measured, for example, by cAMP, IP.sub.3 or intracellular calcium. The invention further relates to a DNA sequence encoding a prostaglandin F2.alpha. receptor, to a recombinant DNA molecule that includes such a DNA sequence and to cells transformed therewith. The invention also relates to antibodies directed against the F2.alpha. receptor and to a method of detecting an F2.alpha. receptor with the antibody. The invention further relates to a method of detecting the presence of an F2.alpha. receptor encoding a DNA fragment in a sample, the use of transformed cells for screening drugs, as well as to drugs prepared using such a screening method.
2. Background Information
Prostaglandin F2.alpha. receptors belong to a large class of hormone receptors which are linked to their signal transduction pathways via guanine nucleotide binding regulatory (G) proteins. Such receptors are amongst the most intensively studied receptor systems. Prostaglandin receptors have been classically defined as being linked to the stimulation of second messengers and measured by cyclic AMP (cAMP), inositol 3-phosphate (IP.sub.3) or intracellular calcium and are coupled with a G regulatory protein (Muallem, Biochem. J. 263: 769-774 (1989)). In contrast, activation of prostaglandin receptors may result in various responses, including inhibition of adenylyl cyclase activity, inhibition of phosphatidylinositol turnover and inhibition of Ca.sup.2+ mobilization (Muallem, Biochem. J. 263: 769-774 (1989), and Duncan, Endocrinology 128: 1519-1526 (1991)). Evidence has also accumulated suggesting heterogeneity in the category of receptors (Balapure, Biol. Reprod. 41: 385-392 (1989)).
Two prostaglandin receptors have previously been cloned, viz. the human and mouse thromboxane A2 receptor and the mouse prostaglandin E.sub.3 receptor (Hirata, Nature 349: 617-620 (1991); Namba, BBRC 184: 1197-1203 (1992); and Sugimoto, J. Biol. Chem. 267: 6463-6466 (1992), respectively).
Prostaglandin F2.alpha. receptors are extremely important from a clinical therapeutic viewpoint. Drugs which activate (agonists) these receptors may be used to treat glaucoma (Alm, Arch. Ophthalmol. 109:1564-1568 (1991)), whereas drugs which block (antagonists) prostaglandin F2.alpha. receptors may be used therapeutically to treat pathological conditions, e.g. in the lungs and uterus. It may be of pharmaceutical value to be able to titer endogenous prostaglandin F2.alpha. with a solubilized receptor as well as to use an immobilized receptor in the purification of a ligand and its analogs. Despite their clinical utility, one problem with the prostaglandin F2.alpha. agonist and putatively antagonist drugs currently available, is that they have many side effects, like many other drugs which work through interaction with receptors. These side effects are predominantly due to a lack of receptor specificity. That is, the drug in use interacts not only with prostaglandin F2.alpha. receptors but with other receptors as well, see e.g. Muallem, Biochem. J. 263; 769-774 (1989).
A major goal of clinical pharmacology and the pharmaceutical industry is the development of more selective drugs with greater efficacy than those currently in use. Impediments to this process are the low abundance of prostaglandin F2.alpha. receptor protein available to study in eye tissue and the lack of suitable homogeneous model systems of the receptors with which to screen drugs against.
SUMMARY OF THE INVENTION
The present invention seeks to provide a solution to this problem by a novel approach which comprises cloning cDNAs encoding prostaglandin F2.alpha. receptors, constructing eukaryotic expression vectors containing these cDNAs, and creating a series of stably transfected mammalian cell lines or prokaryotic cells which express
REFERENCES:
Leung et al., Endocrinology, vol. 119, p. 12, 1986.
Coleman et al., Comprehensive Medicinal Chemistry, vol. 3, pp. 643-659, 1989.
Masu et al., Nature, vol. 329, p. 836, 1987.
Alm, A. et al., PhXA34, a New Potent Ocular Hypotensive Drug "A Study on Dose-Response Relationship and on Aqueous Humor Dynamics in Healthy Volunteers," Arch. Opthalmol. 109: 1564-1568 (1991).
Balapure, A.K. et al., "Multiple Classes of Prostaglandin F.sub.2.alpha. Binding Sites in Subpopulations of Ovine Luteal Cells," Biol. Reprod. 41: 385-392 (1989).
Dohlman H., et al., "A Family of Receptors Coupled to Guanine Nucleotide Regulatory Proteins," Biochem. 26(10):2657-2664 (1987).
Duncan, R. et al., "Prostaglandin F.sub.2.alpha. Stimulates Inositol 1,4,5-Trisphosphate and Inositol 1,3,4,5-Tetrakisphosphate Formation in Bovine Luteal Cells," Endocrinol. 128(3): 1519-1526 (1991).
Hirata, M. et al., "Cloning and Expression of cDNA for a Human Thromboxane A.sub.2 Receptor," Nature 349: 617-620 (1991).
Kozak, M., "Compilation and Analysis of Sequences Upstream from the Translational Start Site in Eukaryotic mRNAs," Nucl. Acids Res. 12(2): 857-872 (1984).
Muallem, S. et al., "Classification of Prostaglandin Receptors Based on Coupling to Signal Transduction Systems," Biochem. J. 263: 769-774 (1989).
Namba, T. et al., "Mouse Thromboxane A.sub.2 Receptor: cDNA Cloning Expression and Northern Blot Analysis," Biochem. Biophys. Res. Comm. 184(3): 1197-1203 (1992).
Orlicky, D.J., et al., "Identification and Purification of a Bovine Corpora Binding Properties," Prostaglandins Leukotrienes and Essential Fatty Acids 41: 51-61 (1990).
Sibley, D. et al., "Regulation of Transmembrane Signaling by Receptor Phosphorylation," Cell 48: 913-922 (1987).
Strader, C. et al., "Structural Basis of .beta.-adrenergic Receptor Function," FASEB J. 3: 1825-1832 (1989).
Sugimoto, Y. et al., "Cloning and Expression of a cDNA for Mouse Prostaglandin E Receptor EP.sub.3 Subtype," J. Biol. Chem. 267(10): 6463-6466 (1992).
Lake Staffan
Stjernschantz Johan
Pharmacia & Upjohn Aktiebolag
Teng Sally P.
LandOfFree
DNA encoding a prostaglandin F28 receptor, a host cell transform does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with DNA encoding a prostaglandin F28 receptor, a host cell transform, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and DNA encoding a prostaglandin F28 receptor, a host cell transform will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-978398