Evolved interferon-alpha polypeptides
View Patent ↗The present invention provides evolved interferon-alpha polypeptides, and conjugates thereof, and nucleic acids encoding the polypeptides. The invention also includes compositions comprising these polypeptides, conjugates, and nucleic acids; cells containing or expressing the polypeptides, conjugates, and nucleic acids; methods of making the polypeptides, conjugates, and nucleic acids; and methods of using the polypeptides, conjugates, and nucleic acids.
1. An isolated or recombinant polypeptide comprising a sequence which differs in 0 to 8 amino acid positions from the sequence SEQ ID NO:312, which polypeptide exhibits antiviral activity.
2. The polypeptide of claim 1 , comprising a sequence which differs in 0 to 6 amino acid positions from the sequence SEQ ID NO:312.
3. The polypeptide of claim 2 , comprising the sequence SEQ ID NO:312.
4. The polypeptide of claim 1 , wherein the antiviral activity of the polypeptide is at least two-fold greater than the antiviral activity of huIFN-alpha 2b or huIFN-alpha 2a in the HeLa/EMCV antiviral assay.
5. A conjugate comprising
(a) the polypeptide of claim 1 ; and
(b) a non-polypeptide moiety covalently attached to the polypeptide.
6. The conjugate of claim 5 , comprising at least two non-polypeptide moieties.
7. The conjugate of claim 5 , comprising a non-polypeptide moiety covalently attached to a cysteine residue.
8. The conjugate of claim 5 , comprising a non-polypeptide moiety covalently attached to a lysine residue or to the N-terminal amino group.
9. The conjugate of claim 5 , comprising a non-polypeptide moiety covalently attached to a lysine residue.
10. The conjugate of claim 5 , comprising a non-polypeptide moiety attached to the N-terminal amino group.
11. The conjugate of claim 5 , wherein the non-polypeptide moiety is a polyethylene glycol.
12. The conjugate of claim 5 , wherein the non-polypeptide moiety is a sugar.
13. The conjugate of claim 5 , wherein the sugar is attached to an N-glycosylation site of the polypeptide.
14. A composition comprising the polypeptide of claim 1 and a pharmaceutically acceptable excipient.
15. A composition comprising the conjugate of claim 5 and a pharmaceutically acceptable excipient.
16. A method for preparing a conjugate, the method comprising
(i) providing the polypeptide of claim 1 , and
(ii) attaching at least one non-polypeptide moiety to the polypeptide, wherein the resulting conjugate exhibits antiviral activity.
17. The method of claim 16 , wherein the step of providing the polypeptide comprises:
providing a culture comprising a host cell, the host cell comprising an expression vector comprising a promoter operably linked to a polynucleotide, the polynucleotide comprising a nucleic acid sequence which encodes the polypeptide,
culturing the culture under conditions which permit expression of the polypeptide, and recovering the polypeptide.
18. The method of claim 17 , wherein the host cell is a eukaryotic host cell or a bacterial host cell.