US 12,331,334 B2
Modified urokinase-type plasminogen activator polypeptides and methods of use
Edwin L. Madison, San Francisco, CA (US); Christopher Thanos, Tiburon, CA (US); Vanessa Soros, San Francisco, CA (US); Mikhail Popkov, San Diego, CA (US); Kimberly Tipton, Boston, MA (US); Matthew John Traylor, Boulder, CO (US); Eric Steven Furfine, Lincoln, MA (US); and Jeffrey Charles Way, Cambridge, MA (US)
Assigned to Vertex Pharmaceuticals, Incorporated, Boston, MA (US)
Filed by Vertex Pharmaceuticals Incorporated, Boston, MA (US)
Filed on Feb. 10, 2023, as Appl. No. 18/167,534.
Application 18/167,534 is a division of application No. 16/734,256, filed on Jan. 3, 2020, granted, now 11,613,744.
Application 16/734,256 is a continuation of application No. PCT/US2019/068839, filed on Dec. 27, 2019.
Claims priority of provisional application 62/786,302, filed on Dec. 28, 2018.
Prior Publication US 2023/0242895 A1, Aug. 3, 2023
Int. Cl. C12N 9/72 (2006.01); A61K 38/00 (2006.01); C07K 14/765 (2006.01)
CPC C12N 9/6462 (2013.01) [C07K 14/765 (2013.01); C12Y 304/21073 (2013.01); A61K 38/00 (2013.01); C07K 2319/02 (2013.01); C07K 2319/50 (2013.01)] 65 Claims
 
1. A nucleic acid molecule, comprising a sequence of nucleotides encoding a modified urokinase-type plasminogen activator (u-PA) polypeptide that comprises one or more amino acid modifications selected from among replacements corresponding to R35Q, R35W, R35Y, H37E, H37Y, V41R, Y40Q, D60aP, L97bA, L97bG, T97aI, H99Q, and conservative amino acid modifications therefor, whereby the modified u-PA polypeptide has increased activity and/or specificity for a complement protein compared to the unmodified active form of the u-PA polypeptide, wherein:
any further amino acid modifications in the encoded modified u-PA polypeptide are selected from among replacements, insertions and deletions in the primary sequence of the modified u-PA polypeptide;
the encoded modified u-PA polypeptide cleaves a complement protein to thereby inhibit or reduce complement activation compared to the unmodified u-PA polypeptide that does not contain the amino acid modifications;
the complement protein is C3;
the encoded modified u-PA polypeptide has reduced activity or specificity for cleavage of a substrate sequence in plasminogen compared to the unmodified u-PA polypeptide;
the encoded modified u-PA polypeptide has at least 90% sequence identity with the polypeptides of any of SEQ ID NOs: 1-6 or a catalytically active portion thereof;
the residues are numbered by chymotrypsin numbering;
the unmodified u-PA polypeptide comprises the sequence set forth in any of SEQ ID NOs: 1-6, or a catalytically active fragment thereof that includes the amino acid modification position(s); and
the conservative modifications are selected from among R35F or N; H37R, Q, W or F; V41K; D60aS; T97aL or V; L97bS; and H99N.