Methods of increasing lean tissue mass using OB protein compositions

• US 7,718,400 B2
• Filed: 03/16/2007
• Issued: 05/18/2010
• Est. Priority Date: 11/22/1995
• Status: Expired due to Fees

• Current Assignee: Amgen Inc.
• Original Assignee: Amylin Pharmaceuticals Incorporated (Bristol-Myers Squibb Company)
• Inventors: Pelleymounter, Mary Ann, Toombs, Christopher Francis, Mann, Michael Benjamin
• Primary Examiner(s): HUYNH, PHUONG N
• Application Number:
  US11/687,591
• Publication Number:
  US 20070224193A1
• Time in Patent Office: 1,159 Days
• Field of Search: 435/69.1, 435/320.1, 435/252.33, 536/23.4
• US Class Current: 435/69.1
• CPC Class Codes: A61K 38/2264 Obesity-gene products, e.g. leptin
• CPC Class Codes: C07K 14/5759 Products of obesity genes, e.g. leptin, obese (OB), tub, fat
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto

Abstract

DNA sequences comprising nucleic acids encoding fusion proteins comprising an Fc portion of an antibody attached at the N-terminus of an OB protein moiety, vectors comprising such DNA sequences, host cells comprising such vectors or DNA sequences, and processes for preparing such fusion proteins, and pharmaceutical compositions comprising such fusion proteins, are described. The DNA sequences, vectors comprising such DNA sequences, host cells comprising such vectors or DNA sequences, and processes for preparing such fusion proteins, and pharmaceutical compositions comprising such fusion proteins are useful, for example, in providing therapeutically effective amounts of compositions useful for, for example, increasing insulin sensitivity, decreasing the dose of insulin required for the treatment of diabetes, controlling serum glucose levels, increasing lean tissue mass, increasing overall body strength, or regulating bone resorption, or effecting any combination of such, in subjects in need or desirous thereof to which the compositions are administered.

Claims

1. A process of producing a fusion protein comprising an Fc portion of an antibody attached at the N-terminus of an OB protein moiety, wherein the process comprises expressing a DNA sequence comprising:

(a) a first nucleic acid sequence encoding said Fc portion of an antibody; and

(b) a second nucleic acid sequence encoding said OB protein moiety selected from the group consisting of;

(i) a nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID. NO;

2 or the amino acid sequence set forth in SEQ ID NO;

4;

(ii) a nucleic acid sequence encoding the amino acid sequence set forth in SEQ. ID. NO;

4 having a lysine residue at position 35 and an isoleucine residue at position 74 according to the position numbering designation set forth in SEQ. ID. NO;

4;

(iii) a nucleic acid sequence encoding the amino acid sequence of subpart (i), lacking a glutaminyl residue at position 28; and

(iv) a nucleic acid sequence encoding the amino acid sequence of subpart (i), wherein the methionyl residue at the N-terminus is absent.

2. The process according to claim 1, wherein said first nucleic acid sequence and said second nucleic acid sequence are attached via a third nucleic acid sequence encoding a linker moiety.

5. An isolated host cell transformed or transfected with the DNA sequence according to claims 1 or 2.

3. A vector comprising a DNA sequence, wherein said DNA sequence comprises:

(a) a first nucleic acid encoding said c portion of an antibody; and

(b) a second nucleic acid sequence encoding said OB protein moiety selected from the group consisting of;

(i) a nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID. NO;

2 or the amino acid sequence set forth in SEQ ID NO;

4;

(ii) a nucleic acid sequence encoding the amino acid sequence set forth in SEQ. ID. NO;

4 having a lysine residue at position 35 and an isoleucine residue at position 74 according to the position numbering designation set forth in SEQ. ID. NO;

4;

(iii) a nucleic acid sequence encoding the amino acid sequence of subpart (i) lacking a glutaminyl residue at position 28; and

(iv) a nucleic acid sequence encoding the amino acid sequence of subpart (i) wherein the methionyl residue at the N-terminus is absent.

4. The vector according to claim 3, wherein said first nucleic acid sequence and said second nucleic acid sequence are attached via a third nucleic acid sequence encoding a linker moiety.

6. An isolated host cell transformed or transfected with the vector according to claim 3 or 4.

7. The host cell according to claim 6, wherein said host cell produces the fusion protein for a use selected from the group consisting of:

increasing insulin sensitivity;

decreasing the dose of insulin required for the treatment of diabetes;

controlling serum glucose levels;

increasing lean tissue mass;

increasing overall body strength; and

regulating bone resorption.

8. The host cell according to claim 7, wherein said host cell produces the fusion protein for use in increasing insulin sensitivity.

9. The host cell according to claim 7, wherein said host cell produces the fusion protein for use in decreasing the dose of insulin required for the treatment of diabetes.

10. The host cell according to claim 7, wherein said host cell produces the fusion protein for use in controlling serum glucose levels.

11. The host cell according to claim 7, wherein said host cell produces the fusion protein for use in increasing lean tissue mass.

12. The host cell according to claim 7, wherein said host cell produces the fusion protein for use in increasing overall body strength.

13. The host cell according to claim 7, wherein said host cell produces the fusion protein for use in regulating bone resorption.