Nucleic acid encoding G-protein coupled receptor with modified DRY motif

• US 7,279,324 B2
• Filed: 01/22/2002
• Issued: 10/09/2007
• Est. Priority Date: 01/23/2001
• Status: Active Grant

• Current Assignee: Duke University
• Original Assignee: Duke University
• Inventors: Caron, Marc G., Oakley, Robert H., Barak, Larry S., Wilbanks, Alyson, Laporte, Stephane A.
• Primary Examiner(s): PAK, MICHAEL D
• Application Number:
  US10/054,616
• Publication Number:
  US 20030049643A1
• Time in Patent Office: 2,086 Days
• Field of Search: None
• US Class Current: 435/320.1
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A61K 31/41 having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
• CPC Class Codes: A61K 31/64 Sulfonylureas, e.g. glibenclamide, tolbutamide, chlorpropamide
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 11/08 Bronchodilators
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/22 Anxiolytics
• CPC Class Codes: A61P 25/24 Antidepressants
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 27/06 Antiglaucoma agents or miotics
• CPC Class Codes: A61P 27/16 Otologicals
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 37/08 : Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 : Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/06 : Antiarrhythmics
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61P 9/12 : Antihypertensives
• CPC Class Codes: C07K 14/705 : Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 14/723 : G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH receptor

Abstract

The present invention relates to modified G-protein coupled receptors (GPCRs). The modified GPCRs of the present invention include GPCRs that have been modified to have altered DRY motifs such that the modified GPCRs are constitutively desensitized. As such, the modified GPCRs of the present invention preferably localize to endocytic vesicles or endosomes in an agonist-independent manner. The invention also relates to methods of screening compounds and sample solutions for GPCR activity using the modified GPCRs.

Claims

1. A nucleic acid selected from the group consisting of SEQ ID NOs:

7–

12.

3. A vector comprising a nucleic acid of claim 1.

4. An isolated host cell comprising the expression vector of claim 3.

2. A nucleic acid encoding a polypeptide selected from the group consisting of SEQ ID NOs.:

1–

6.

5. A nucleic acid encoding a modified GPCR or biologically active fragment thereof comprising a DRY motif modified to contain an amino acid other than arginine at position 2, wherein the modified GPCR or biologically active fragment thereof is constitutively desensitized in the absence of an agonist, wherein said nucleic acid is at least 80% identical to a nucleic acid selected from the group consisting of SEQ ID NOs:

7–

12.

6. A nucleic acid encoding a modified GPCR or biologically active fragment thereof comprising a DRY motif modified to contain an amino acid other than arginine at position 2, wherein the modified GPCR or biologically active fragment thereof is constitutively desensitized in the absence of an agonist, wherein said nucleic acid is at least 90% identical to a nucleic acid selected from the group consisting of SEQ ID NOs:

7–

12.

7. A nucleic acid encoding a modified GPCR or biologically active fragment thereof comprising a DRY motif modified to contain an amino acid other than arginine at position 2, wherein the modified GPCR or biologically active fragment thereof is constitutively desensitized in the absence of an agonist, wherein said nucleic acid is at least 95% identical to a nucleic acid selected from the group consisting of SEQ ID NOs:

7–

12.

8. A nucleic acid encoding a modified GPCR or biologically active fragment thereof comprising a DRY motif modified to contain an amino acid other than arginine at position 2, wherein the modified GPCR or biologically active fragment thereof is constitutively desensitized in the absence of an agonist, wherein said nucleic acid is at least 98% identical to a nucleic acid selected from the group consisting of SEQ ID NOs:

7–

12.

9. A nucleic acid encoding a modified GPCR or biologically active fragment thereof comprising a DRY motif modified to contain an amino acid other than arginine at position 2, wherein the modified GPCR or biologically active fragment thereof is constitutively desensitized in the absence of an agonist, wherein said nucleic acid is at least 99% identical to a nucleic acid selected from the group consisting of SEQ ID NOs:

7–

12.