Methods for identifying compounds useful in treating type II diabetes

• US 5,795,726 A
• Filed: 01/10/1997
• Issued: 08/18/1998
• Est. Priority Date: 11/12/1996
• Status: Expired due to Fees

• Current Assignee: Millennium Pharmaceuticals Incorporated (Takeda Pharmaceutical Company Limited)
• Original Assignee: Millennium Pharmaceuticals Incorporated (Takeda Pharmaceutical Company Limited)
• Inventors: Glucksmann, M. Alexandra
• Primary Examiner(s): Saunders, David
• Assistant Examiner(s): VANDERVEGT, FRANCOIS P
• Application Number:
  US08/782,047
• Time in Patent Office: 585 Days
• Field of Search: 435/4, 435/6, 435/8, 435/7.21, 536/23.5
• US Class Current: 435/7.21
• 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: C07K 14/70571 for neuromediators, e.g. serotonin receptor, dopamine receptor

Abstract

Methods for identifying compounds, which modulate the bioactivity of human hepatic nuclear factor-1 (HNF-1), and which are therefore useful in treating type II diabetes are disclosed.

Claims

1. A method for aiding in the identification of a compound for use in treating Type II diabetes comprising the steps of:

a) incubating a cell in the presence and absence of a test compound, wherein said cells been altered to contain a nucleic acid molecule comprising an HNF1 response element operably linked to a reporter gene and said cell expresses a wild-type HNF1 protein;

b) determining the level of expression of said reporter gene in said cell incubated in the presence and absence of said test compound; and

c) selecting a compound that induces a higher level of expression of said reporter gene in step (b) than that expressed in the absence of said compound, for use in treating Type II diabetes.

2. The method of claim 1, wherein said HNF1 response element is contained in a promoter selected from the group consisting of:

liver specific glucokinase, glucose transporter, pyruvate kinase, albumin, aldolase B, UDP-N-acetylglucosamine, an alcohol dehydrogenase, cytochrome P-450, aminopeptidase N, phenylalaninehydroxylase, tyrosine aminotransferase, hepatic triglyceride lipase, apolipoprotein B, apolipoprotein A-II, cholesterol 7 alpha-hydrolase, fibrinogen, alpha-1-microglobulin, C4b-binding protein insulin like growth factor 1 (IGF-1), alpha-fetoprotein, and neural cell adhesion molecule (N-CAM).

3. The method of claim 1, wherein said HNF1 response element is contained in an albumin promoter.

4. The method of claim 1, wherein said cell further expresses a mutant HNF1 protein selected from the group consisting of:

an HNF1 protein encoded by a nucleic acid molecule having a base substitution at nucleotide 414 which results in a substitution of an arginine by a tryptophan in the wild-type HNF1 amino acid sequence and an HNF1 protein encoded by a nucleic acid molecule having a nucleotide insertion at nucleotide 895 resulting in a frameshift mutation at residue 895.

5. The method of claim 2, wherein said cell further expresses a mutant HNF1 protein selected from the group consisting of:

an HNF1 protein encoded by a nucleic acid molecule having a base substitution at nucleotide 414 which results in a substitution of an arginine by a tryptophan in the wild-type HNF1 amino acid sequence and an HNF1 protein encoded by a nucleic acid molecule having a nucleotide insertion at nucleotide 895 resulting in a frameshift mutation at residue 895.

6. The method of claim 2, wherein said reporter gene is a luciferase gene.

7. The method of claim 3, wherein said reporter gene is a luciferase gene.

8. The method of claim 4, wherein said reporter gene is a luciferase gene.

9. The method of claim 5, wherein said reporter gene is a luciferase gene.

10. A method for identifying a compound for use in treating Type II diabetes comprising the steps of:

a) incubating a pancreatic cell in the presence and absence of a test compound, wherein said cell has been altered to contain a nucleic acid molecule which encodes HNF1 and said cell produces insulin;

b) determining the level of insulin expression in said cell incubated in the presence and absence of said test compound; and

c) selecting a compound that induces a higher level of insulin expression than that expressed in the absence of said compound for use in treating Type II diabetes.