Compounds and methods for purification of serine proteases

Patent metadata

Field	Value
Publication number	US-9644195-B2
Application number	US-201514723395-A
Country	US
Kind code	B2
Filing date	May 27, 2015
Priority date	Dec 27, 2012
Publication date	May 9, 2017
Grant date	May 9, 2017

Abstract

Disclosed herein are compounds, compositions, methods and kits for purifying a serine protease and serine proteases purified with the compounds, compositions and methods.

First claim

What is claimed is:

1. A compound selected from:

or a salt thereof.

2. An affinity solid support selected from:

or a salt thereof,
wherein:
Y—Z is

Z is a solid support;
q is 3, 4 or 6; and
v is 1, 2, 3, 4, 5, 6 or 7.

3. The affinity solid support of claim 2 , wherein Z is an agarose resin.

4. A kit for purifying a serine protease comprising
(1) an affinity solid support of claim 2 , and
(2) an elution buffer comprising a competitive agent.

5. A kit for purifying a serine protease comprising a compound of claim 1 and a resin of the formula NC—O—Z or

wherein Z is a solid support.

6. The kit of claim 5 , wherein Z is an agarose resin.

7. A method for purifying a fXa derivative comprising
(1) adding a first composition comprising the fXa derivative to an affinity solid support of claim 2 or a salt thereof to form a second composition comprising the fXa derivative and the affinity solid support of claim 2 , and
(2) eluting the fXa derivative from the second composition with an elution buffer comprising a competitive agent,
wherein the fXa derivative is a polypeptide comprising the amino acid sequence of SEQ ID NO: 1, 2 or 4, or a polypeptide having at least about 80% sequence identity to SEQ ID NO: 1, 2 or 4 and further wherein the derivative is capable of binding to a fXa inhibitor and does not assemble into a prothrombinase complex.

8. The method of claim 7 , wherein the competitive agent is arginine and/or benzamidine, or a salt thereof.

9. The method of claim 7 , wherein the fXa derivative is a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, or a polypeptide having at least about 90% sequence identity to SEQ ID NO: 2.

10. The method of claim 9 , wherein the polypeptide has at least about 95% sequence identity to SEQ ID NO: 2.

Assignees

• Portola Pharm Inc

Inventors

• Pandey Anjali
• Rose Jack W

Classifications

• C07D213/72

  Nitrogen atoms (nitro radicals C07D213/61) · CPC title

• B01J20/286

  Phases chemically bonded to a substrate, e.g. to silica or to polymers · CPC title

• B01J2220/80

  Aspects related to sorbents specially adapted for preparative, analytical or investigative chromatography · CPC title

• B01J20/267

  Cross-linked polymers · CPC title

• C07D213/75

  Amino or imino radicals, acylated by carboxylic or carbonic acids, or by sulfur or nitrogen analogues thereof, e.g. carbamates · CPC title