# US 2013/0156848 A1 · Compositions and methods for modulating hemostasis

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- Number: US 2013/0156848 A1
- Document ID: US20130156848A1
- Status: Published application
- Kind: A1
- Application number: 13/710,046
- Filed: 2012-12-10
- Published: 2013-06-20
- Earliest claimed: 2005-11-15
- Assignees: The United States of America National Institutes of Health (NIH), U.S. Dept. of Health and Human Services (DHHS), NIH Division of Extramural Inventions and Technology Resources (DEITR)
- Inventors: Sriram Krishnaswamy; Elsa P. Bianchini; Steven Orcutt
- Web page: https://patentgrep.com/patent/US20130156848A1

## CPC

- C12N 9/6429: {Thrombin (3.4.21.5)}
- C12Y 304/21005: Thrombin (3.4.21.5)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Y: ENZYMES

## Abstract

Novel thrombin/prothrombin protease/zymogen variants which have anticoagulation activity and methods of use thereof are disclosed.

## Claims

### Claim at position 1

1-17. (canceled)

### Claim 18

A nucleic acid encoding a variant thrombin zymogen/protease which modulates hemostasis.

### Claim 19 (depends on 1)

The nucleic acid of claim 1, wherein said variant zymogen is a thrombin variant having the sequence of SEQ ID NO: 1, wherein each X at the thrombin B chain protease domain start can be any amino acid.

### Claim 20 (depends on 19)

The nucleic acid of claim 19, wherein said encoded variant contains at least one modification in SEQ ID NO: 1 selected from the group consisting of: a) Ile at position 321 is Thr, or Val; b) Val at position 322 is Ala, or Ile, or Glu and c) Glu at position 323 is Thr, or Gly.

### Claim 21 (depends on 20)

A nucleic acid as claimed in claim 20 wherein the nucleotides at positions 1122-1131 of SEQ ID NO: 2 encode amino acids selected from the group consisting of Thr-Ala-Thr, Val-Ile-Glu and Ile-Glu-Gly.

### Claim 22 (depends on 21)

The nucleic acid of claim 21 cloned into an expression vector.

### Claim 23 (depends on 22)

The vector of claim 22, selected from the group consisting of an adenoviral vector, an adenovirus-associated vector, a retroviral vector, a plasmid, and a lentiviral vector.

### Claim 24 (depends on 21)

A method for treatment of a hemostasis related disorder in a patient in need thereof comprising administration of an expression vector encoding a therapeutically effective amount of the zymogen/protease thrombin variant of claim 21 in a biologically acceptable carrier.

### Claim 25 (depends on 24)

The method of claim 24, wherein said variant is an anticoagulant and said disorder is selected from the group consisting of thrombosis, thrombocytopenia, stroke, and coagulapathy.

### Claim 26 (depends on 24)

The method of claim 24, wherein said the amino acids at positions 321, 322 and 323 in SEQ ID NO: 2 are selected from the group consisting of Thr-Ala-Thr, Val-Ile-Glu and Ile-Glu-Gly.

### Claim 27 (depends on 24)

The method of claim 24, wherein said the amino acids at positions 321, 322, 323 in SEQ ID NO: 2 are selected from the group consisting of Val-Ile-Glu and Ile-Glu-Gly.

### Claim 28 (depends on 24)

The method of claim 24, wherein said expression vector is encapsulated in a liposome or mixed with phospholipids or micelles.

### Claim 29 (depends on 22)

A host cell expressing the vector of claim 22.