Hedgehog fusion proteins
AssigneeCuris, Inc.
InventorsKathryn Strauch, Ellen A. Garber, Frederick R. Taylor
Life of this patent
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Abstract
A fusion polypeptide is described having the amino acid sequence X-Y-Z, or portion thereof, comprising the amino acid sequence of a hedgehog (X); Y is an optional linker moiety; and Z is a polypeptide comprising at least a portion of a polypeptide other than hedgehog. It is preferred that X is a human hedgehog. Mutants of hedgehog are also described.
Claims39
An isolated polypeptide having an amino acid sequence X-Y-Z, wherein
- X is a polypeptide comprising the amino acid sequence of a Sonic hedgehog protein, wherein the Sonic hedgehog protein comprises:
- a) an N-terminal 20 kDa fragment of a full-length naturally-occurring Sonic hedgehog sequence, wherein a KEX2 recognition site corresponding to residues 32-36 of a human Shh protein (SEQ ID NO:15) is replaced with a sequence selected from one of SEQ ID NO:88-94 or 99-102 or
- b) an N-terminal 20 kDa fragment of a full-length naturally-occurring Sonic hedgehog sequence wherein the N-terminal cysteine is substituted with phenylalanine, isoleucine, methionine, or two isoleucine residues, and wherein a KEX2 recognition site corresponding to residues 32-36 of a human Shh protein (SEQ ID NO:15) is replaced with a sequence selected from one of SEQ ID NO:88-94 or 99-102;
- Y is an optional linker moiety;
- and
- Z is a polypeptide comprising at least a portion of a polypeptide other than hedgehog.
A fusion protein having an amino terminal-region consisting of the amino acid sequence of a Sonic hedgehog protein and having a carboxy terminal region comprising at least a portion of a protein other than hedgehog, wherein the Sonic hedgehog protein comprises:
- a) an N-terminal 20 kDa fragment of a full-length naturally-occurring Sonic hedgehog sequence, wherein a KEX2 recognition site corresponding to residues 32-36 of a human Shh protein (SEQ ID NO:15) is replaced with a sequence selected from one of SEQ ID NO:88-94 or 99-102 or
- b) an N-terminal 20 kDa fragment of a full-length naturally-occurring Sonic hedgehog sequence wherein the N-terminal cysteine is substituted with phenylalanine, isoleucine, methionine, or two isoleucine residues, and wherein a KEX2 recognition site corresponding to residues 32-36 of a human Shh protein (SEQ ID NO:
- 15) is replaced with a sequence selected from one of SEQ ID NO:
- 88-94 or 99-102.
An isolated polypeptide having the amino acid sequence X-Y-Z, wherein
- X is a polypeptide having the amino acid sequence of a Sonic hedgehog protein, wherein the Sonic hedgehog protein comprises an N-terminal 20 kDa fragment of a full-length naturally-occurring Sonic hedgehog sequence wherein the N-terminal cysteine is substituted with phenylalanine, isoleucine, methionine, or two isoleucine residues, and wherein a KEX2 recognition site corresponding to residues 32-36 of a human Shh protein (SEQ ID NO:15) is replaced with a sequence selected from one of SEQ ID NO:88-94 or 99-102;
- Y is an optional linker moiety;
- and
- Z is a polypeptide comprising at least a portion of a constant region of an immunoglobulin.
A fusion protein having an amino terminal region consisting of the amino acid sequence of a Sonic hedgehog protein and having a carboxy terminal region comprising at least a portion of a constant region of an immunoglobulin, wherein the Sonic hedgehog protein comprises an N-terminal 20 kDa fragment of a full-length naturally-occurring Sonic hedgehog sequence wherein the N-terminal cysteine is substituted with phenylalanine, isoleucine, methionine, or two isoleucine residues, and wherein a KEX2 recognition site corresponding to residues 32-36 of a human Shh protein (SEQ ID NO:15) is replaced with a sequence selected from one of SEQ. ID NO:88-94 or 99-102.
An isolated polypeptide having an amino acid sequence X-Y-Z, wherein X is a polypeptide comprising the amino acid sequence of a Sonic hedgehog protein, wherein the Sonic hedgehog protein comprises:
- a) an N-terminal 20 kDa fragment of a full-length naturally-occurring Sonic hedgehog sequence, wherein a KEX2 recognition site corresponding to residues 32-36 of a human Shh protein (SEQ ID NO:15) is replaced with the sequence of SEQ ID NO:
- 92 or
- b) an N-terminal 20 kDa fragment of a full-length naturally-occurring Sonic hedgehog sequence wherein the N-terminal cysteine is substituted with phenylalanine, isoleucine, methionine, or two isoleucine residues, and wherein a KEX2 recognition site corresponding to residues 32-36 of a human Shh protein (SEQ ID NO:15) is replaced with the sequence of SEQ ID NO:
- 92;
- Y is an optional linker moiety;
- and
- Z is a polypeptide comprising at least a portion of a polypeptide other than hedgehog.
The isolated polypeptide of 30, wherein X is a hedgehog agonist that binds patched-1 with an affinity that is similar to or higher than the binding affinity of a mature naturally-occurring hedgehog protein to patched-1.
A fusion protein having an amino terminal region consisting of the amino acid sequence of a Sonic hedgehog protein and having a carboxy terminal region comprising at least a portion of a protein other than hedgehog, wherein the Sonic hedgehog protein comprises:
- a) an N-terminal 20 kDa fragment of a full-length naturally-occurring Sonic hedgehog sequence, wherein a KEX2 recognition site corresponding to residues 32-36 of a human Shh protein (SEQ ID NO:15) is replaced with the sequence of SEQ ID NO:
- 92 or
- b) an N-terminal 20 kDa fragment of a full-length naturally-occurring Sonic hedgehog sequence wherein the N-terminal cysteine is substituted with phenylalanine, isoleucine, methionine, or two isoleucine residues, and wherein a KEX2 recognition site corresponding to residues 32-36 of a human Shh protein (SEQ ID NO:
- 15) is replaced with the sequence of SEQ ID NO:
- 92.
A isolated polypeptide having the amino acid sequence X-Y-Z, wherein X is a polypeptide having the amino acid sequence of a Sonic hedgehog protein, wherein the Sonic hedgehog protein comprises an N-terminal 20 kDa fragment of a full-length naturally-occurring Sonic hedgehog sequence wherein the N-terminal cysteine is substituted with phenylalanine, isoleucine, methionine, or two isoleucine residues, and wherein a KEX2 recognition site corresponding to residues 32-36 of a human Shh protein (SEQ ID NO:15) is replaced with the sequence of SEQ ID NO:
- 92;
- Y is an optional linker moiety;
- and
- Z is a polypeptide comprising at least a portion of a constant region of an immunoglobulin.
A fusion protein having an amino terminal region consisting of the amino acid sequence of a Sonic hedgehog protein and having a carboxy terminal region comprising at least a portion of a constant region of an immunoglobulin, wherein the Sonic hedgehog protein comprises an N-terminal 20 kDa fragment of a full-length naturally-occurring Sonic hedgehog sequence wherein the N-terminal cysteine is substituted with phenylalanine, isoleucile, methiodine, or two isoleucine residues, and wherein a KEX2 recognition site corresponding to residues 32-36 of a human Shh protein (SEQ ID NO:15) is replaced with the sequence of SEQ ID NO:
- 92.
Dependencies are inferred from the claim text.