Therapeutic uses of modified relaxin polypeptides
Modified relaxin polypeptides and their uses, including therapeutic uses thereof for the treatment of a fibrotic disorder or heart failure are provided. Exemplary embodiments provide for the use of relaxin polypeptides which include one or more amino acid substitutions with natural or non-naturally encoded amino acids, and/or linkage to a water-soluble polymer, such as polyethylene glycol.
1. A method of treating a patient having a fibrotic disorder, comprising administering to said patient an effective amount of a biologically active modified relaxin polypeptide, wherein (a) said modified relaxin polypeptide comprises an A chain and a B chain linked by at least one disulfide bond, wherein said A chain comprises SEQ ID NO: 4 substituted with a non-naturally encoded amino acid at position 1 and said B chain comprises SEQ ID NO:5 or SEQ ID NO:6, and (b) the non-naturally encoded amino acid is linked to a linker, polymer, or biologically active molecule, wherein said non-naturally encoded amino acid comprises a first functional group and the linker, polymer, or biologically active molecule comprises a second functional group, wherein the first functional group and second functional group are not identical and each comprise a carbonyl group, an aminooxy group, a hydrazide group, a hydrazine group, a semicarbazide group, an azide group, or an alkyne group, and the non-naturally encoded amino acid is linked to the linker, polymer, or biologically active molecule by the resultant covalent linkage created by the reaction of the first and second functional groups.
2. The method of claim 1 , wherein the non-naturally encoded amino acid at position 1 of the A chain is linked to a water soluble polymer.
3. The method of claim 2 wherein said water soluble polymer comprises a polyethylene glycol (PEG).
4. The method of claim 3 wherein said PEG has an average molecular weight of about 20 kDa.
5. The method of claim 1 wherein said non-naturally encoded amino acid comprises a para-substituted phenylalanine, ortho-substituted phenylalanine, or meta-substituted phenylalanine.
6. The method of claim 1 wherein said non-naturally encoded amino acid comprises para-acetyl-L-phenylalanine.
7. The method of claim 1 , wherein the fibrotic disorder is selected from cardiac fibrosis, pulmonary fibrosis, renal fibrosis, hepatic fibrosis, coronary fibrosis, bone marrow fibrosis, dermatological fibrosis, and a fibrotic eye disorder.
8. A method of treating heart failure in a patient in need thereof, comprising administering to said patient an effective amount of a biologically active modified relaxin polypeptide, said modified relaxin polypeptide comprising an A chain and a B chain linked by at least one disulfide bond, wherein said A chain comprises SEQ ID NO: 4 substituted with a non-naturally encoded amino acid at position 1 and said B chain comprises SEQ ID NO:5 or SEQ ID NO:6, wherein the non-naturally encoded amino acid at position 1 of the A chain is linked to a water soluble polymer, and said non-naturally encoded amino acid comprises para-acetyl-L-phenylalanine.
9. The method of claim 8 , wherein said non-naturally encoded amino acid is linked to said water soluble polymer by an oxime linkage produced by the reaction of a carbonyl moiety and aminooxy (hydroxylamine) moiety, or by a triazole linkage produced by the reaction of an azide moiety and an alkyne moiety.
10. The method of claim 8 , wherein the heart failure comprises congestive heart failure.
11. The method of claim 8 , wherein the heart failure comprises chronic heart failure.
12. The method of claim 8 , wherein the heart failure comprises acute heart failure.