Peptides and compositions for treatment of joint damage
The present invention provides new protease resistant polypeptides, as well as compositions and methods for treating, ameliorating or preventing conditions related to joint damage, including acute joint injury and arthritis.
1. An isolated polynucleotide encoding a polypeptide having at least 95% amino acid sequence identity to a polypeptide consisting of the amino acid sequence set forth as SEQ ID NO: 28, wherein the polypeptide comprises an amino acid that is a polar amino acid other than K or R at position 423 as determined with reference to SEQ ID NO:1, and wherein the polypeptide has chondrogenic activity.
2. The isolated polynucleotide according to claim 1 , which is complementary DNA (cDNA) or messenger RNA (mRNA).
3. The isolated polynucleotide according to claim 1 , wherein the encoded polypeptide further comprises a secretion signal sequence.
4. The isolated polynucleotide according to claim 1 , wherein the encoded polypeptide further comprises a fusion domain selected from the group consisting of a polyhistidine, Glu-Glu, glutathione S transferase (GST), thioredoxin, protein A, protein G, an immunoglobulin heavy chain constant region (Fc), maltose binding protein (MBP), a human serum albumin (HSA), a FLAG tag, influenza virus haemagglutinin (HA), and a c-myc tag.
5. The isolated polynucleotide according to claim 1 , which is codon-optimized for expression in a host cell.
6. The isolated polynucleotide according to claim 1 , which is operably linked to a promoter or other regulatory sequence.
7. A cloning or expression vector comprising one or more polynucleotides according to claim 1 , wherein the vector is suitable for the recombinant production of the encoded polypeptide.
8. A host cell, comprising one or more cloning or expression vectors according to claim 7 .
9. The host cell of claim 8 , which is a mammalian host cell, a bacterial host cell, a yeast host cell, or an insect host cell.
10. A process for the production of a polypeptide, comprising culturing a host cell according to claim 9 under conditions sufficient to express the polypeptide, and thereafter purifying and recovering the polypeptide from the host cell culture.
11. A kit comprising the polynucleotide according to claim 1 , wherein the kit comprises: a first container comprising the polynucleotide; and a second container having an aqueous reconstitution formula.
12. The kit according to claim 11 , wherein one container comprises a single chambered pre-filled syringe.
13. The kit according to claim 11 , wherein the containers are encompassed as a multi-chambered pre-filled syringe.