High-affinity antagonists of ELR-CXC chemokines
AssigneeUniversity of Saskatchewan Technologies Inc.
InventorsJohn R. Gordon, Fang Li
Life of this patent
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Abstract
The present invention provides novel nucleic acids, novel polypeptide sequences encoded by these nucleic acids, methods for production thereof, and uses thereof, for a novel ELR-CXC chemokine receptor antagonist.
Claims79
An isolated polynucleotide comprising a sequence selected from the group comprising:
- (i) the nucleotide sequence of SEQ ID NO:4;
- (ii) a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1;
- (iii) a nucleotide sequence encoding an ELR-CXC chemokine antagonist comprising an amino acid sequence substantially equivalent to a wild-type bovine CXCL8 sequence, wherein
- (a) the amino acid sequence of the antagonist starts at amino acid residue Thr3,
- (b) Lys11 of the wild-type sequence is substituted with Arg in the amino acid sequence of the antagonist, and
- (c) Gly31 of the wild-type sequence is substituted with Pro in the in the amino acid sequence of the antagonist;
- (iv) a nucleotide sequence encoding an ELR-CXC chemokine antagonist comprising an amino acid sequence substantially equivalent to a wild-type bovine CXCL8 sequence, wherein
- (a) the amino acid sequence of the antagonist starts at amino acid residue Thr3,
- (b) Lys11 of the wild-type sequence is substituted with Arg in the amino acid sequence of the antagonist,
- (c) Gly31 of the wild-type sequence is substituted with Pro in the in the amino acid sequence of the antagonist, and
- (d) Pro32 of the wild-type sequence is substituted with Gly in the amino acid sequence of the antagonist;
- and
- (v) a nucleotide sequence encoding an ELR-CXC chemokine antagonist comprising an amino acid sequence substantially equivalent to a wild-type bovine CXCL8 sequence, wherein
- (a) the amino acid sequence of the antagonist starts at amino acid residue Thr3,
- (b) Lys11 of the wild-type sequence is substituted with Arg in the amino acid sequence of the antagonist,
- (c) Thr12 of the wild-type sequence is substituted with Ser in the amino acid sequence of the antagonist,
- (d) His13 of the wild-type sequence is substituted with Phe in the amino acid sequence of the antagonist, and
- (e) Gly31 of the wild-type sequence is substituted with Pro in the in the amino acid sequence of the antagonist.
An isolated polynucleotide encoding a polypeptide capable of high-affinity binding to CXCR1 and CXCR2 receptors in a mammalian inflammatory cell so as to inhibit activation of the inflammatory cell.
An ELR-CXC chemokine antagonist comprising an amino acid sequence substantially equivalent to a wild-type bovine CXCL8 sequence, said amino acid sequence further comprising differences from the wild-type amino acid sequence selected from the group comprising:
- (i) a truncation of the first two amino acid residues of the antagonist sequence such that it starts at amino acid residue Thr3, a substitution of Lys11 of the wild-type sequence with Arg in the amino acid sequence of the antagonist, and a substitution of Gly31 of the wild-type sequence with Pro in the amino acid sequence of the antagonist;
- (ii) a truncation of the first two amino acid residues of the antagonist sequence such that it starts at amino acid residue Thr3, a substitution of Lys11 of the wild-type sequence with Arg in the amino acid sequence of the antagonist, a substitution of Gly31 of the wild-type sequence with Pro in the in the amino acid sequence of the antagonist, and a substitution of Pro32 of the wild-type sequence with Gly in the amino acid sequence of the antagonist;
- and
- (iii) a truncation of the first two amino acid residues of the antagonist sequence such that it starts at amino acid residue Thr3, a substitution of Lys11 of the wild-type sequence with Arg in the amino acid sequence of the antagonist, a substitution of Thr12 of the wild-type sequence with Ser in the amino acid sequence of the antagonist, a substitution of His13 of the wild-type sequence with Phe in the amino acid sequence of the antagonist, and a substitution of Gly31 of the wild-type sequence with Pro in the in the amino acid sequence of the antagonist.
A method of producing the polypeptide of 4 in an animal comprising:
- (i) introducing a gene encoding said polypeptide into said animal;
- (ii) growing said animal;
- (iii) accumulating said polypeptide in said animal;
- (iv) preparing an extract containing said polypeptide;
- (v) purifying said polypeptide.
A method of producing the polypeptide of 6 in an animal comprising:
- (i) introducing a gene encoding said polypeptide into said animal;
- (ii) growing said animal;
- (iii) accumulating said polypeptide in said animal;
- (iv) preparing an extract containing said polypeptide;
- (v) purifying said polypeptide.
A gene fusion comprising an affinity handle and a polynucleotide comprising a sequence selected from the group comprising:
- (i) the nucleotide sequence of SEQ ID NO:4;
- (ii) a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1;
- (iii) a nucleotide sequence encoding an ELR-CXC chemokine antagonist comprising an amino acid sequence substantially equivalent to a wild-type bovine CXCL8 sequence, wherein
- (a) the amino acid sequence of the antagonist starts at amino acid residue Thr3,
- (b) Lys11 of the wild-type sequence is substituted with Arg in the amino acid sequence of the antagonist, and
- (c) Gly31 of the wild-type sequence is substituted with Pro in the in the amino acid sequence of the antagonist;
- (iv) a nucleotide sequence encoding an ELR-CXC chemokine antagonist comprising an amino acid sequence substantially equivalent to a wild-type bovine CXCL8 sequence, wherein
- (a) the amino acid sequence of the antagonist starts at amino acid residue Thr3,
- (b) Lys11 of the wild-type sequence is substituted with Arg in the amino acid sequence of the antagonist,
- (c) Gly31 of the wild-type sequence is substituted with Pro in the in the amino acid sequence of the antagonist, and
- (d) Pro32 of the wild-type sequence is substituted with Gly in the amino acid sequence of the antagonist;
- and
- (v) a nucleotide sequence encoding an ELR-CXC chemokine antagonist comprising an amino acid sequence substantially equivalent to a wild-type bovine CXCL8 sequence, wherein
- (a) the amino acid sequence of the antagonist starts at amino acid residue Thr3,
- (b) Lys11 of the wild-type sequence is substituted with Arg in the amino acid sequence of the antagonist,
- (c) Thr12 of the wild-type sequence is substituted with Ser in the amino acid sequence of the antagonist,
- (d) His13 of the wild-type sequence is substituted with Phe in the amino acid sequence of the antagonist, and
- (e) Gly31 of the wild-type sequence is substituted with Pro in the in the amino acid sequence of the antagonist.
A gene fusion comprising an affinity handle and a polynucleotide encoding a polypeptide capable of high-affinity binding to CXCR1 and CXCR2 receptors in a mammalian inflammatory cell so as to inhibit activation of the inflammatory cell.
An ELR-CXC chemokine antagonist comprising an amino acid sequence substantially equivalent to a wild-type bovine CXCL8 sequence, said amino acid sequence further comprising differences from the wild-type amino acid sequence selected from the group comprising:
- (i) a truncation of the first two amino acid residues of the antagonist sequence such that it starts at amino acid residue Thr3, a substitution of Lys11 of the wild-type sequence with Arg in the amino acid sequence of the antagonist, and a substitution of Gly31 of the wild-type sequence with Pro in the amino acid sequence of the antagonist;
- (ii) a truncation of the first two amino acid residues of the antagonist sequence such that it starts at amino acid residue Thr3, a substitution of Lys11 of the wild-type sequence with Arg in the amino acid sequence of the antagonist, a substitution of Gly31 of the wild-type sequence with Pro in the in the amino acid sequence of the antagonist, and a substitution of Pro32 of the wild-type sequence with Gly in the amino acid sequence of the antagonist;
- and
- (iii) a truncation of the first two amino acid residues of the antagonist sequence such that it starts at amino acid residue Thr3, a substitution of Lys11 of the wild-type sequence with Arg in the amino acid sequence of the antagonist, a substitution of Thr12 of the wild-type sequence with Ser in the amino acid sequence of the antagonist, a substitution of His13 of the wild-type sequence with Phe in the amino acid sequence of the antagonist, and a substitution of Gly31 of the wild-type sequence with Pro in the in the amino acid sequence of the antagonist.
A method of producing the fusion polypeptide of 40 in an animal comprising:
- (i) introducing the gene fusion into said animal;
- (ii) growing said animal;
- (iii) accumulating said fusion polypeptide in said animal;
- (iv) preparing an extract containing said fusion polypeptide;
- (v) purifying said fusion polypeptide.
A method of producing the fusion polypeptide of 41 in an animal comprising:
- (i) introducing the gene fusion into said animal;
- (ii) growing said animal;
- (iii) accumulating said fusion polypeptide in said animal;
- (iv) preparing an extract containing said fusion polypeptide;
- (v) purifying said fusion polypeptide.
A pharmaceutical composition comprising a biologically-active amount of one of the novel polypeptides are preferred embodiments of the invention.
A compound having a three dimensional structure, wherein the three dimensional structure is capable of high-affinity binding to CXCR1 and CXCR2 receptors in a mammalian inflammatory cell so as to inhibit activation of the inflammatory cell.
Dependencies are inferred from the claim text.