Beta-amyloid peptide-binding proteins and polynucleotides encoding the same

• US 20020058267A1
• Filed: 05/09/2001
• Published: 05/16/2002
• Est. Priority Date: 04/16/1997
• Status: Active Grant

• Current Assignee: Wyeth (Pfizer Inc.)
• Original Assignee: American Home Products Corporation (Pfizer Inc.)
• Inventors: Ozenberger, Bradley A., Howland, David, Walker, Stephen G., Bard, Jonathan A., Sofia, Heidi, Kajkowski, Eileen M., Jacobsen, Jack S.
• Granted Patent: US 6,787,319 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2217/075 inducing loss of function, i.e. knock out
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• 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 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 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: C07K 14/4711 Alzheimer's disease; Amyloid plaque core protein
• CPC Class Codes: C07K 2319/00 Fusion polypeptide

Abstract

Novel proteins which bind human β-amyloid peptide, polynucleotides which encode these proteins, and methods for producing these proteins are provided. Diagnostic, therapeutic, and screening methods employing the polynucleotides and polypeptides of the present invention are also provided. Transgenic animals and knockout animals are also provided.

Claims

1. A polynucleotide comprising an endogenous variant of the nucleotide sequence of SEQ ID NO:

1, or a degenerate variant of said endogenous variant.

2. A polynucleotide according to claim 1 further comprising a splice variant.

20. A diagnostic process comprising analyzing for the presence of a polynucleotide of claim 1 in a sample derived from a host.

3. A polynucleotide encoding a β

-amyloid peptide-binding (BBP) protein comprising a PXDGS motif beginning at amino acid 237.

4. A protein comprising the amino acid of SEQ ID NO:

2.

5. A protein comprising the amino acid of SEQ ID NO:

2 from amino acid 68 to amino acid 269.

6. A protein comprising the amino acid sequence encoded by the cDNA insert of clone BBP1-fl deposited under accession number ATCC 98617.

7. A protein comprising the amino acid sequence from amino acid 185 to amino acid 217 of SEQ ID NO:

2.

8. A fusion protein comprising a BBP1 linked to a heterologous protein or peptide sequence.

9. The fusion protein of claim 8 wherein the BBP1 has the amino acid sequence of SEQ ID NO:

2.

10. A method for determining a polynucleotide encoding a β

-amyloid peptide-binding protein (BBP) in a sample comprising the steps of (a) hybridizing to a sample a probe specific for said polynucleotide under conditions effective for said probe to hybridize specifically to said polynucleotide; and

(b) determining the hybridization of said probe to polynucleotides in the sample, wherein said probe comprises a nucleic acid sequence having a region of 20 or more base pairs at least 90% identical to the polynucleotide sequence of SEQ ID NO;

1.

11. A method for determining a polynucleotide encoding a β

-amyloid peptide-binding protein (BBP) in a sample comprising the steps of (a) hybridizing to a sample a probe specific for said polynucleotide under conditions effective for said probe to hybridize specifically to said polynucleotide; and

(b) determining the hybridization of said probe to polynucleotides in the sample, wherein said probe comprises a nucleic acid sequence having a region of 20 or more base pairs at least 30% identical to the polynucleotide sequence of the cDNA insert of ATCC 98617 or ATCC 98399.

15. An antibody according to claim 14 wherein the extracellular region comprises the PXDGS motif.

12. An antibody that binds specifically to a polypeptide comprising the amino acid sequence of SEQ ID NO:

1.

13. An antibody that binds specifically to a polypeptide comprising the amino acid sequence of the β

-amyloid peptide binding protein encoded by the cDNA insert of ATCC 98617.

14. An antibody that binds to an extracellular region of a BBP.

16. A method for detecting in a sample a polypeptide comprising a region at least 90% identical to the amino acid sequence of SEQ ID NO:

2, said method comprising (a) incubating with a sample a reagent that bind specifically to said polypeptide under conditions effective for specific binding; and

(b) determining the binding of said reagent to said polypeptide in the sample.

17. A method for detecting in a sample a polypeptide comprising a region at least 90% identical in sequence to the amino acid sequence of the β

-amyloid peptide binding protein encoded by the cDNA insert of ATCC 98617, said method comprising (a) incubating with a sample a reagent that bind specifically to said polypeptide under conditions effective for specific binding; and

(b) determining the binding of said reagent to said polypeptide in the sample.

18. A method for diagnosing a disease characterized by aberrant expression of human β

-amyloid peptide (BAP), comprising (a) incubating a sample indicative of the aberrant expression of human β

-amyloid peptide with a reagent comprising a polypeptide comprising a region at least 90% identical to the amino acid sequence of SEQ ID NO;

2 under conditions effective for specific binding of said reagent to said human β

-amyloid peptide; and

(b) determining the binding of said reagent to said human β

-amyloid peptide in the sample.

19. A method for diagnosing a disease characterized by aberrant expression of human β

-amyloid peptide, comprising (a) incubating a sample indicative of the aberrant expression of human β

-amyloid peptide with a reagent comprising a polypeptide comprising a region at least 90% identical to the amino acid sequence of the β

-amyloid peptide binding protein encoded by the cDNA insert of ATCC 98617 under conditions effective for specific binding of said reagent to said human β

-amyloid peptide; and

(b) determining the binding of said reagent to said human β

-amyloid peptide in the sample.

21. A method for identifying compounds which regulate the activity of a β

-amyloid peptide binding protein comprising (a) incubating a sample comprising human β

-amyloid peptide in a test medium containing said test compound and a reagent comprising a polypeptide comprising a region at least 90% identical to the amino acid sequence of SEQ ID NO;

2 under conditions effective for specific binding of said reagent to said human β

-amyloid peptide;

(b) comparing the binding of said reagent to said peptide in the sample in the presence and absence of said test compound; and

(c) relating the difference between the binding in step (b) to the test compound regulating the activity of the β

-amyloid peptide binding protein.

22. A method for identifying compounds which regulate the activity of a β

-amyloid peptide binding protein comprising (a) incubating a sample comprising human β

-amyloid peptide in a test medium containing said test compound and a reagent comprising a polypeptide comprising a region at least 90% identical to the amino acid sequence of the β

-amyloid peptide binding protein encoded by the cDNA insert of ATCC 98617 under conditions effective for specific binding of said reagent to said human β

-amyloid peptide;

(b) comparing the binding of said reagent to said peptide in the sample in the presence and absence of said test compound; and

(c) relating the difference between the binding in step (b) to the test compound regulating the activity of the β

-amyloid peptide binding protein.

26. The animal of claim 25 wherein the transgene is under the control of a regulatable expression system.

29. A knockout according to claim 28 wherein the Cre gene is under the control of a tissue specific promoter.

30. A knockout according to claim 28 wherein the Cre gene is under the control of a developmentally specific promoter.

31. A knockout according to claim 28 wherein the Cre gene is under the control of an inducible promoter.

23. A method for the treatment of a patient having need to inhibit β

-amyloid peptide accumulation in the brain comprising administering to the patient a therapeutically effective amount of BBP1.

24. A method for the treatment of a patient having need of such treatment comprising administering to the patient a therapeutically effective amount of an antibody which binds to an extracellular portion of BBP1.

25. A transgenic or chimeric nonhuman animal comprising the polynucleotide of SEQ ID NO:

1 or a degenerate variant of said polynucleotide.

27. A knockout nonhuman animal wherein at least one allele of the BBP1 gene has been functionally disrupted.

28. A knockout nonhuman animal wherein at least one allele of the BBP1 gene can be functionally disrupted by the induction of the Cre gene.

32. A method for inhibiting expression of the BBP1 gene comprising providing to a cell double stranded ribonucleic acid complementary to a portion of the BBP1 gene wherein said ribonucleic acid comprises about 600 base pairs.

33. A method of inhibiting expression of the BBP1 gene in a cell comprising providing said cell with an antisense nucleic acid.