Recombinantly modified plasmin

• US 8,034,913 B2
• Filed: 04/21/2005
• Issued: 10/11/2011
• Est. Priority Date: 04/22/2004
• Status: Expired due to Fees

• Current Assignee: Grifols Therapeutics LLC (Grifols SA)
• Original Assignee: Grifols Therapeutics Incorporated (Grifols SA)
• Inventors: Hunt, Jennifer A., Novokhatny, Valery
• Primary Examiner(s): Nguyen; Quang
• Application Number:
  US11/568,023
• Publication Number:
  US 20100304465A1
• Time in Patent Office: 2,364 Days
• Field of Search: None
• US Class Current: 536/23.2
• CPC Class Codes: C12N 9/6435 Plasmin (3.4.21.7), i.e. fibrinolysin
• CPC Class Codes: C12Y 304/21007 Plasmin (3.4.21.7), i.e. fibrinolysin

Abstract

Polynucleotides and polypeptides relating to a recombinantly-modified plasmin(ogen) molecule are provided. The plasmin(ogen) molecule has a single kringle domain N-terminal to the activation site present in the native human plasminogen molecule, and exhibits lysine-binding and significant enzymatic characteristics associated with the native enzyme.

Claims

1. A polynucleotide comprising a nucleotide sequence encoding a polypeptide having a single N-terminal kringle domain which is at least 90% identical to the kringle 1 domain of native human plasminogen;

directly attached to a C-terminal domain which is at least 90% identical to the activation site and serine protease domain of native human plasminogen, wherein the polypeptide binds to immobilized lysine and can be activated by a plasminogen activator.

2. The polynucleotide of claim 1, wherein the encoded polypeptide is SEQ ID NO:

2 with no more than 30 amino acid substitutions.

3. The polynucleotide of claim 1, wherein the encoded polypeptide is at least 90% identical to the sequence shown in SEQ ID NO:

2.

4. The polynucleotide of claim 1, wherein the encoded polypeptide is at least 95% identical to the sequence shown in SEQ ID NO:

2.

5. The polynucleotide of claim 1, wherein the encoded polypeptide is the sequence shown in SEQ ID NO:

2.

6. The polynucleotide of claim 1, wherein the nucleotide sequence of the polynucleotide is the sequence shown in SEQ ID NO:

1 or a degenerate variant that encodes SEQ ID NO;

2.

7. The polynucleotide of claim 1, wherein the polypeptide exhibits a fibrinolytic activity that is inhibited by α

2-antiplasmin at a rate of inhibition that is at least about 5-fold faster than the rate of inhibition of the fibrinolytic activity of mini-plasmin by α

2-antiplasmin.

8. An expression vector comprising the polynucleotide of claim 1.

9. A cultured cell comprising the expression vector of claim 8.

10. The polynucleotide of claim 1, wherein the encoded polypeptide is at least 98% identical to the sequence shown in SEQ ID NO:

2.