	US 7,547,509 B2
	Cyanovirin variant-polymer conjugates
M. Elizabeth Snell, Ardmore, Ala. (US); Michael J. Roberts, Williamsburg, Va. (US); Toshiyuki Mori, Germantown, Md. (US); Barry R. O'Keefe, Frederick, Md. (US); and Michael R. Boyd, Mobile, Ala. (US)
Assigned to Nektar Therapeutics AL, Corporation, Huntsville, Ala. (US); and National Institute of Heatlh, Washington, D.C. (US)
Filed on Aug. 02, 2007, as Appl. No. 11/832,925.
Application 11/832925 is a division of application No. 10/742465, filed on Dec. 18, 2003, granted, now 7,267,941.
Claims priority of provisional application 60/461731, filed on Apr. 09, 2003.
Claims priority of provisional application 60/435950, filed on Dec. 19, 2002.
Prior Publication US 2008/0015151 A1, Jan. 17, 2008
Int. Cl. C12Q 1/70 (2006.01); C12P 21/06 (2006.01); C12P 21/04 (2006.01); A61B 5/055 (2006.01); A61K 39/00 (2006.01)

U.S. Cl. 435—5 [435/68.1; 435/71.1; 424/9.34; 424/134.1]

10 Claims

1. A method for preparing an antiviral polypeptide-polymer conjugate, comprising:

(i) providing an antiviral polypeptide having at least 70% sequence identity to native cyanovirin-N (SEQ ID NO. 1), wherein said polypeptide is modified relative to SEQ ID NO: 1 to contain 1-4 reactive sites for selective conjugation, each said reactive site being selected from:

(a) a cysteine substitution or insertion of at least one position selected from the group consisting of 5, 9-21, 25, 29-40, 45-49, 52, 57, 59-72, 79-91, 96-101 of SEQ ID NO: 1, the C-terminus, and the N-terminus; and

(b) a single lysine residue remaining after arginine substitution of all but one of the lysine residues in the polypeptide;

or a fragment thereof, wherein the fragment includes the sequence set forth in residues 40-80 of SEQ ID NO: 1, modified to contain at least one said reactive site for selective conjugation; and

(ii) covalently attaching thereto, at at least one said reactive site for selective conjugation, a water-soluble polymer,

wherein said antiviral polypeptide-polymer conjugate has antiviral activity.