Pegylated porcine interferon and methods of use thereof
Disclosed herein are porcine interferon alpha variants (pIFN-α) comprising a synthetic amino acid at select locations in pIFN-α and a one or two amino acid insertion in the N-terminus after removal of the signal peptide. The pIFN-α variants can further be pegylated. Methods of making and administering these compounds to treat virus infections in pigs and formulations comprising the variants are also provided.
1. A porcine interferon-α (pIFN-α) variant comprising:
(SEQ ID NO: 1)
X a X b CDLPQTHSLAHTRALRLLAQMRRISPFSCLDHRRDFGSPHEAFGGN
QVQKAQAMALVHEMLQQTFQLFSTEGSAAAWNESLLHQFYTGLDQQLRDL
EACVMQEAGL E GTPLLEEDSIRAVRKYFHRLTLYLQEKSYSPCAWEIVRA
EVMRSFSSSRNLQDRLRKKE,
wherein residue E103, E107, L112, Y136, or Q102 (numbering with respect to SEQ ID NO: 4) is substituted with a synthetic amino acid; and
wherein X a X b are variable positions selected from the group consisting of no amino acids, a single proline residue, and a proline-serine dipeptide.
2. The pIFN-α variant of claim 1 , wherein the synthetic amino acid is para-acetyl-phenylalanine (pAF).
3. The pIFN-α variant of claim 1 , wherein the variant is selected from the group consisting of SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 19, and SEQ ID NO: 20.
4. The pIFN-α variant of claim 1 , wherein the synthetic amino acid is pegylated.
5. The pIFN-α variant of claim 4 , wherein the pegylated pIFN-α variant is pegylated with about a 5 kDa to 40 kDa PEG.
6. The pIFN-α variant of claim 5 , wherein the PEG is a 30 kDa PEG.
7. The pIFN-α variant of claim 1 in a formulation comprising 20 mM sodium acetate, 100 mM sodium chloride, 5% glycerol at pH 5.0 of about 2.0 to about 6.0 g/L titer of pIFN-α variant.
8. A porcine interferon-α (pIFN-α) variant consisting of:
(SEQ ID NO: 14)
PSCDLPQTHSLAHTRALRLLAQMRRISPFSCLDHRRDFGSPHEAFGGNQV
QKAQAMALVHEMLQQTFQLFSTEGSAAAWNESLLHQFYTGLDQQLRDLEA
CVMQEAGL-pAF-GTPLLEEDSIRAVRKYFHRLTLYLQEKSYSPCAWEIV
RAEVMRSFSSSRNLQDRLRKKE,
wherein a residue corresponding to E107 (numbering with respect to SEQ ID NO: 4) is substituted with para-acetyl-phenylalanine (pAF) and said pAF residue is pegylated with a 30 kDa linear PEG.
9. A method of treating a virus infection in a pig comprising administering subcutaneously to said pig in need thereof a pIFN-α variant; wherein the pIFN-α variant is selected from the group consisting of SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEO ID NO: 18, SEQ ID NO: 19, and SEQ ID NO: 20.
10. The method of claim 9 , wherein the pIFN-α variant is administered in the range of 25 μg/kg to 150 μg/kg animal weight.
11. The method of claim 10 , comprising a second administration of 25 μg/kg to 150 μg/kg animal weight of said pIFN-α variant.
12. The method of claim 11 , wherein the second administration is 7 to 14 days after first administration.
13. The method of claim 9 , wherein the virus infection is selected from the group consisting of: porcine reproductive and respiratory syndrome virus, foot and mouth disease virus, swine influenza virus, porcine circovirus, porcine epidemic diarrhea virus and transmissible gastroenteritis virus.
14. The method of claim 9 , wherein the pig is a newborn pig or the pig is a pregnant pig.