Materials and methods relating to endothelial cell growth inhibitors
AssigneeKarolinska Innovations AB
InventorYihai Cao
Life of this patent
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Abstract
Proteolytic enzymes are involved in generation of a number of endogenous angiogenesis inhibitors and it has been shown that urokinase-activated plasmin can process plasminogen to release an angiogenesis inhibitor, K1–5 (protease-activated kringles 1–5). However, this proteolytic protein (proteolytic K1–5) cannot be secreted (exported) from cells and thus is limited in its application as a therapeutic. The present inventor has now engineered a recombinant protein derived from plasminogen which is folded correctly such that it can be secreted from cells in which it is expressed. The inventor surprisingly found that in order to fold correctly such that it can be secreted, the recombinant protein must comprise a secretory signal peptide and a pre-activation peptide derived from plasminogen.
Claims25
A pharmaceutical composition comprising a recombinant protein having the amino acid sequence of Kringle 1 to 5, a secretory signal peptide and a pre-activation peptide, which are the Kringle 1 to 5, secretory signal peptide and pre-activation peptide of a mammalian plasminogen, wherein said protein has anti-angiogenic activity, and wherein the amino acid sequence of Kringle 1 to 5 consists of the sequence of amino acids 97–562 of SEQ ID NO:
- 5.
A pharmaceutical composition comprising a recombinant protein consisting essentially of having the amino acid sequence of that of Kringle 1 to 5, a secretory signal peptide and a pre-activation peptide, which are the Kringle 1 to 5, secretory signal peptide and pre-activation peptide of a mammalian plasminogen, wherein said protein has anti-angiogenic activity, and wherein the amino acid sequence of Kringle 1 to 5 consists of the sequence of amino acids 97–562 of SEQ ID NO:
- 5.
A recombinant protein comprising having the amino acid sequence of Kringle 1–5, a secretory signal peptide and a preactivation peptide, which are the Kringle 1 to 5, secretory signal peptide and pre-activation peptide of a mammalian plasminogen, wherein the amino acid sequence of Kringle 1 to 5 consists of the sequence of amino acids 97–562 of SEQ ID NO:
- 5, and wherein the preactivation peptide comprises the sequence of amino acids 33–96 of SEQ ID NO. 5, and wherein the protein has anti-angiogenic activity.
A recombinant protein comprising having the amino acid sequence of Kringle 1–5, a secretory signal peptide and a preactivation peptide, which are the Kringle 1 to 5, secretory signal peptide and preactivation peptide of a mammalian plasminogen, wherein the amino acid sequence of Kringle 1 to 5 consists of the sequence of amino acids 97–562 of SEQ ID NO:
- 5, and wherein the secretory signal peptide comprises the sequence of amino acids 1–32 of SEQ ID NO. 5, and wherein the protein has anti-angiogenic activity.
A recombinant protein comprising the amino acid sequence of Kringle 1–5, a secretory signal peptide, an a preactivation peptide, which are the Kringle 1 to 5, secretory signal peptide and a preactivation peptide of a mammalian plasminogen, wherein the recombinant protein further comprises a tumour targeting polypeptide and wherein said tumour targeting peptide comprises an amino acid sequence selected from the group consisting of CNGRCVSGCAGCR (SEQ ID NO:
- 7) and CDRGDCFC (SEQ ID NO:
- 9).
Dependencies are inferred from the claim text.