Interferon-Alpha Polypeptides and Conjugates
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 .- 36 . (canceled)
37 . An isolated or recombinant polypeptide comprising a sequence which differs in 0 to 16 amino acid positions from a sequence selected from SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, 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:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:74, SEQ ID NO:75, SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:90, SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:99, SEQ ID NO:100, SEQ ID NO:101, SEQ ID NO:102, SEQ ID NO:103, and SEQ ID NO:104, which polypeptide exhibits antiviral activity.
38 . The polypeptide of claim 1 , comprising a sequence which differs from SEQ ID NO:84 in 0 to 16 amino acid positions.
39 . The polypeptide of claim 38 , comprising a sequence which differs from SEQ ID NO:84 in 0 to 8 amino acid positions.
40 . The polypeptide of claim 37 , wherein the antiviral activity of the polypeptide is equal to or greater than the antiviral activity of huIFN-alpha 2b or huIFN-alpha 2a.
41 . The polypeptide of claim 37 , 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.
42 . A conjugate comprising
(a) the polypeptide of claim 37 ; and
(b) a non-polypeptide moiety covalently attached to the polypeptide.
43 . The conjugate of claim 42 , comprising at least two non-polypeptide moieties.
44 . The conjugate of claim 42 , comprising a non-polypeptide moiety covalently attached to a lysine residue.
45 . The conjugate of claim 42 , wherein the non-polypeptide moiety is a polymer.
46 . The conjugate of claim 45 , wherein the polymer is a polyethylene glycol.
47 . The conjugate of claim 42 , wherein the non-polypeptide moiety is a sugar.
48 . The conjugate of claim 47 , wherein the sugar is attached to an N-glycosylation site.
49 . A composition comprising the polypeptide of claim 37 and a pharmaceutically acceptable excipient.
50 . A composition comprising the conjugate of claim 42 and a pharmaceutically acceptable excipient.
51 . An isolated or recombinant nucleic acid comprising a polynucleotide sequence which encodes the polypeptide of claim 37 .
52 . A host cell comprising the nucleic acid of claim 51 .
53 . A vector comprising the nucleic acid of claim 51 .
54 . A method for preparing a polypeptide, the method comprising:
providing a culture comprising a host cell, the host cell comprising an expression vector comprising a promoter operably linked to a nucleic acid, the nucleic acid comprising a polynucleotide sequence which encodes the polypeptide of claim 37 ,
culturing the culture under conditions which permit expression of the polypeptide, and recovering the polypeptide.
55 . The method of claim 54 , wherein the host cell is a eukaryotic host cell or a bacterial host cell.
56 . A method for preparing a conjugate, the method comprising
(i) providing the polypeptide of claim 37 , and
(ii) attaching at least one non-polypeptide moiety to an attachment group of the polypeptide, wherein the resulting conjugate exhibits antiviral activity.
57 . The method of claim 56 , 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 nucleic acid, the nucleic acid comprising a polynucleotide sequence which encodes the polypeptide,
culturing the culture under conditions which permit expression of the polypeptide, and recovering the polypeptide.
58 . The method of claim 57 , wherein the host cell is a eukaryotic host cell or a bacterial host cell.
59 . A method for reducing the level of a virus in the serum of a patient infected with the virus, comprising administering to the patient the polypeptide of claim 37 in an amount effective to reduce the level of the virus in the serum compared to the level present prior to the start of treatment.
60 . A method for reducing the level of a virus in the serum of a patient infected with the virus, comprising administering to the patient the conjugate of claim 42 in an amount effective to reduce the level of the virus in the serum compared to the level present prior to the start of treatment.