Interferon-alpha polypeptides and conjugates
View Patent ↗The present invention provides interferon-alpha polypeptides and conjugates, 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
(a) differs in 0 to 8 amino acid positions from SEQ ID NO:13;
(b) comprises Ala or Leu at position 48; and
(c) optionally comprises a methionine at the N-terminus;
which polypeptide exhibits antiviral activity.
2. The polypeptide of claim 1 , which sequence comprises Ala at position 48.
3. An isolated or recombinant polypeptide comprising the sequence SEQ ID NO:13, optionally further comprising a methionine at the N-terminus, which polypeptide exhibits antiviral activity.
4. The polypeptide of claim 1 or 3 , wherein the antiviral activity of the polypeptide is equal to or greater than the antiviral activity of huIFN-alpha 2b or huIFN-alpha 2a.
5. The polypeptide of claim 4 , 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.
6. The polypeptide of claim 1 or 3 , wherein the polypeptide exhibits a ratio of antiviral activity/antiproliferative activity at least two-fold greater than the ratio of antiviral activity/antiproliferative activity exhibited by huIFN-alpha 2b or huIFN-alpha 2a.
7. A composition comprising the polypeptide of claim 1 or 3 , and a pharmaceutically acceptable excipient.
8. A method for preparing a conjugate, the method comprising
(i) providing the polypeptide of claim 1 or 3 , and
(ii) attaching at least one non-polypeptide moiety to the polypeptide, wherein the resulting conjugate exhibits antiviral activity.
9. The method of claim 8 , wherein the step of attaching the at least one non-polypeptide moiety to the polypeptide comprises:
reacting the polypeptide with 40 kDa mPEG2-NHS at a molar ratio of mPEG2-NHS:polypeptide of about 3:1 to 5:1 at pH 9 and 4° C. for 1 hour to overnight.
10. The polypeptide of claim 1 , comprising a sequence which differs from SEQ ID NO:13 in 0 to 4 amino acid positions.
11. A conjugate comprising
(a) the polypeptide of claim 1 or 3 , and
(b) a non-polypeptide moiety covalently attached to the polypeptide,
which conjugate exhibits antiviral activity.
12. The conjugate of claim 11 , comprising at least two non-polypeptide moieties covalently attached to the polypeptide.
13. The conjugate of claim 11 , comprising a non-polypeptide moiety covalently attached to a cysteine residue.
14. The conjugate of claim 11 , comprising a non-polypeptide moiety covalently attached to a lysine residue or to the N-terminal amino group.
15. The conjugate of claim 11 , comprising a non-polypeptide moiety covalently attached to a lysine residue.
16. The conjugate of claim 11 , comprising a non-polypeptide moiety attached to the N-terminal amino group.
17. The conjugate of claim 11 , wherein the non-polypeptide moiety is a polymer.
18. The conjugate of claim 17 , wherein the polymer is a polyethylene glycol.
19. The conjugate of claim 18 , wherein the polyethylene glycol is a 40 kDa mPEG2 moiety covalently attached to a lysine residue.
20. The conjugate of claim 11 , wherein the non-polypeptide moiety is a sugar.
21. The conjugate of claim 20 , wherein the sugar is attached to an N-glycosylation site of the polypeptide.
22. A conjugate comprising
(a) a polypeptide comprising the sequence SEQ ID NO:13, optionally further comprising a methionine at the N-terminus; and
(b) a PEG moiety covalently attached to a lysine residue of the polypeptide, which conjugate exhibits antiviral activity.
23. The conjugate of claim 22 , wherein the PEG moiety is a 40 kDa mPEG2 moiety.
24. The conjugate of claim 23 , wherein the 40 kDa mPEG2 moiety is covalently attached to a lysine residue selected from Lys122 and Lys135.
25. A composition comprising the conjugate of claim 22 , and a pharmaceutically acceptable excipient.
26. A composition comprising
(a) a conjugate comprising a polypeptide comprising the sequence SEQ ID NO:13, optionally further comprising a methionine at the N-terminus, and a 40 kDa mPEG2 moiety covalently attached to Lys122; and
(b) a conjugate comprising a polypeptide comprising the sequence SEQ ID NO:13, optionally further comprising a methionine at the N-terminus, and a 40 kDa mPEG2 moiety covalently attached to Lys135;
which composition exhibits antiviral activity.
27. The composition of claim 26 further comprising a pharmaceutically acceptable excipient.