Synthetic chemokine receptor ligands and methods of use thereof
The present invention provides synthetic CXCR3 ligands, including consensus CXCR3 ligands and hybrid CXCR3 ligands; and compositions comprising the ligands. The present invention provides polynucleotides encoding the synthetic CXCR3 ligands; expression vectors comprising the polynucleotides; and host cells comprising the polynucleotidess. The present invention provides methods of treating fibrotic disorders; methods of treating angiogenic disorders; methods of treating cancer; and methods of treating bacterial infections. The methods generally involve administering to an individual in need thereof an effective amount of a subject synthetic CXCR3 ligand. (SEQ ID NO: 15) MKKSGVLFLLGIILLVLIGVQGFPMFKRGRCLCIGPGVKPVNPRSLEKLE IIPASQFCPRIEIIATLKNGVQTCLNPDSKQARLIIKKVSKEMSKRSP
1 . A synthetic CXCR3 polypeptide ligand comprising a polypeptide of from about 70 to about 125 amino acids in length, optionally further including an additional methionine attached to the ordinarily first amino acid at the N-terminus, the amino acid sequence of the polypeptide comprising, in sequence, discrete sub-sequences corresponding in amino acid identity and number to sub-sequences of different, naturally occurring CXCR3 ligands selected from IP-10, I-TAC, and Mig, where the amino acid sequence of the synthetic CXCR3 polypeptide differs from the amino acid sequence of naturally occurring CXCR3 ligands IP-10, I-TAC, and Mig.
2 . The synthetic CXCR3 polypeptide ligand of claim 1 , wherein the CXCR3 ligand comprises the amino acid sequence as set forth in any one of SEQ ID NOs:15-20.
3 . A synthetic CXCR3 ligand comprising a polypeptide of from about 70 to about 125 amino acids in length, optionally further including an additional methionine attached to the ordinarily first amino acid at the N-terminus, the amino acid sequence of the polypeptide comprising those amino acid residues that are common to IP-10, Mig, and I-TAC, and which comprises, at one or more of those positions where there is no amino acid common to IP-10, Mig, and I-TAC, an amino acid which predominantly occurs at that position.
4 . The synthetic CXCR3 polypeptide ligand of claim 3 , wherein the CXCR3 ligand comprises the amino acid sequence as set forth in any one of SEQ ID NO:01, 02, and 03.
5 . A composition comprising the synthetic CXCR3 ligand of any of claims 1 - 4 .
6 . A polynucleotide comprising a nucleotide sequence encoding a synthetic CXCR3 ligand of any of claims 1 - 4 .
7 . The polynucleotide of claim 6 , wherein-said synthetic CXCR3 ligand comprises the amino acid sequence set forth in any one of SEQ ID NO:01, 02, 03, 15, 16, 17, 18, 19, and 20.
8 . An expression vector comprising the polynucleotide of claim 6 operably linked to a promoter.
9 . A host cell comprising the polynucleotide of claim 6 .
10 . A host cell comprising the expression vector of claim 8 .
11 . A method for producing a synthetic CXCR3 ligand, the method comprising:
culturing the host cell of claim 10 under conditions that favor production of the synthetic CXCR3 ligand; and
isolating the synthetic CXCR3 ligand from the culture.
12 . An antibody that specifically binds a synthetic CXCR3 ligand of any one of claims 1 - 4 .
13 . A method of treating a fibrotic disease in an individual, the method comprising administering to an individual suffering from a fibrotic disease an amount of a synthetic CXCR3 ligand that is effective in the treatment or prophylaxis of the fibrotic disease in the individual.
14 . The method of claim 13 , wherein the fibrotic disease is pulmonary fibrosis.
15 . The method of claim 13 , wherein the pulmonary fibrosis is idiopathic pulmonary fibrosis.
16 . The method of claim 13 , wherein the pulmonary fibrosis is from a known etiology.
17 . The method of claim 13 , wherein the fibrotic disease is selected from liver fibrosis, renal fibrosis, cardiac fibrosis, and scleroderma.
18 . A method of reducing tumor growth in an individual having a tumor, the method comprising administering to the individual an effective amount of a synthetic CXCR3 ligand.
19 . The method of claim 18 , further comprising administering an effective amount of an anti-neoplastic agent selected from an alkylating agent, a nitrosourea, an antimetabolite, an antitumor antibiotic, a plant (vinca) alkaloid, a taxane, and a steroid hormone.
20 . The method of any of claims 13 - 19 , wherein the individual is a human.