Modified and Stabilized GDF Propeptides and Uses Thereof

• US 20120252884A1
• Filed: 06/14/2012
• Published: 10/04/2012
• Est. Priority Date: 02/08/2001
• Status: Active Grant

• Current Assignee: Wyeth LLC (Pfizer Inc.)
• Original Assignee: Wyeth LLC (Pfizer Inc.)
• Inventors: Wolfman, Neil M., Khor, Soo-Peang, Tomkinson, Kathleen N.
• Granted Patent: US 8,710,025 B2
• Time in Patent Office: Days
• US Class Current: 514/44.R
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 19/10 for osteoporosis
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 21/06 Anabolic agents androgens A61P5/26
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/475 Growth factors; Growth regulators
• CPC Class Codes: C07K 14/51 Bone morphogenetic factor; Osteogenins; Osteogenic factor; Bone-inducing factor
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 2319/00 Fusion polypeptide

Abstract

Modified and stabilized propeptides of Growth Differentiation Factor proteins, such as GDF-8 and Bone Morphogenetic Protein-11, are disclosed. Also disclosed are methods for making and using the modified propeptides to prevent or treat human or animal disorders in which an increase in muscle tissue would be therapeutically beneficial. Such disorders include muscle or neuromuscular disorders (such as amyotrophic lateral sclerosis, muscular dystrophy, muscle atrophy, congestive obstructive pulmonary disease, muscle wasting syndrome, sarcopenia, or cachexia), metabolic diseases or disorders (such as such as type 2 diabetes, noninsulin-dependent diabetes mellitus, hyperglycemia, or obesity), adipose tissue disorders (such as obesity), and bone degenerative diseases (such as osteoporosis).

Claims

1-118. -118. (canceled)

119. A nucleic acid molecule encoding a GDF-8 propeptide comprising a nucleic acid sequence that is at least 95% identical to SEQ ID NO:

6 and encodes a propeptide that is modified at the aspartate residue corresponding to aspartate 76 of SEQ ID NO;

5.

122. A pharmaceutical composition comprising the nucleic acid of claim 119 and a pharmaceutically acceptable carrier or vector.

123. A vector comprising the nucleic acid of claim 119.

125. The vector of claim 123 further comprising a nucleic acid that encodes an Fc region of an IgG molecule.

126. The vector of claim 125 further comprising a linker peptide between the nucleotides encoding the GDF-8 propeptide and the IgG Fc region.

127. The vector of claim 126, wherein the linker peptide comprises nucleotides encoding for the amino acid sequence consisting of glycine-serine-glycine-serine (GSGS) (SEQ ID NO:

17).

129. The method of making a GDF-8 propeptide that has been modified at the aspartate residue corresponding to aspartate 76 of SEQ ID NO:

5 comprising;

(a) preparing a nucleic acid molecule according to claim 119;

(b) preparing a nucleic acid molecule encoding the Fc region of the IgG molecule; and

(c) fusing the nucleic acid molecules from steps (a) and (b) to produce a modified GDF-8 propeptide.

130. The method of claim 129, further comprising preparing a double stranded oligonucleotide encoding a linker peptide and fusing the cDNA molecules from steps (a) and (b) to either end of the double stranded oligonucleotide encoding the linker peptide.

131. The method of claim 130, wherein the linker peptide comprises the amino acid sequence consisting of glycine-serine-glycine-serine (GSGS).

132. The pharmaceutical composition of claim 122, wherein the vector is a viral vector.

133. The vector of claim 123, wherein the vector is a viral vector.

120. A nucleic acid molecule encoding a GDF-8 propeptide comprising a nucleic acid that hybridizes under stringent hybridization conditions (hybridization at 65°

in 6×

SSC) to the complement of SEQ ED NO;

6 and encodes a propeptide that is modified at the aspartate residue corresponding to aspartate 76 of SEQ ID NO;

5, wherein the encoded propeptide inhibits at least one GDF-8 biological activity.

121. The nucleic acid of claim 120, wherein the GDF-8 propeptide negatively regulates skeletal muscle mass.

124. A vector comprising the nucleic acid of claim 120.

128. A host cell comprising the vector of claim 120.