# US 2019/0169267 A1 · Factor viii complex with xten and von willebrand factor protein, and uses thereof

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- Number: US 2019/0169267 A1
- Document ID: US20190169267A1
- Status: Published application
- Kind: A1
- Application number: 16/154,310
- Filed: 2018-10-08
- Published: 2019-06-06
- Earliest claimed: 2012-07-11
- Assignees: Bioverativ Therapeutics Inc.
- Applicants: Bioverativ Therapeutics Inc.
- Inventors: Ekta Seth CHHABRA; Tongyao LIU; Pei-yun CHANG; Robert T. PETERS; John KULMAN
- Web page: https://patentgrep.com/patent/US20190169267A1

## CPC

- C07K 14/755: Factors VIII {, e.g. factor VIII C (AHF), factor VIII Ag (VWF)}
- A61K 38/36: Blood coagulation or fibrinolysis factors
- A61K 38/37: Factors VIII
- A61K 47/55: the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug
- A61P 7/04: Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- C07K 16/00: Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
- C12N 15/11: DNA or RNA fragments; Modified forms thereof (DNA or RNA not used in recombinant technology, C07H21/00); {Non-coding nucleic acids having a biological activity}
- A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00)
- C07K 2319/00: Fusion polypeptide
- C07K 2319/30: Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
- C07K 2319/31: fusions, other than Fc, for prolonged plasma life, e.g. albumin
- C07K 2319/35: containing a fusion for enhanced stability/folding during expression, e.g. fusions with chaperones or thioredoxin
- C07K 2319/50: containing protease site
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present invention provides a chimeric protein comprising a VWF protein comprising the D′ domain and D3 domain of VWF, one or more XTEN sequence, and a FVIII protein, wherein the VWF fragment, the XTEN sequence, or the FVIII protein are linked to or associated with each other. The chimeric protein can further comprise one or more Ig constant region or a portion thereof (e.g., an Fc region). A polypeptide chain comprising a VWF fragment of the invention binds to or is associated with a polypeptide chain comprising a FVIII protein linked to an XTEN sequence and the polypeptide chain comprising the VWF fragment can prevent or inhibit binding of endogenous VWF to the FVIII protein linked to the XTEN sequence. By preventing or inhibiting binding of endogenous VWF to the FVIII protein, which is a half-life limiting factor for FVIII, the VWF fragment can induce extension of half-life of the chimeric protein comprising a FVIII protein. The invention also includes nucleotides, vectors, host cells, methods of using the VWF fragment, or the chimeric proteins.

## Claims

### Claim at position 1

1-119. (canceled)

### Claim 120

A polynucleotide or a set of polynucleotides encoding a chimeric protein comprising (i) a von Willebrand Factor (VWF) protein comprising the D′ domain and the D3 domain of VWF, (ii) an XTEN sequence, and (iii) a FVIII protein, wherein the VWF fragment and the XTEN sequence are linked by an optional linker, wherein the VWF fragment or the XTEN sequence is linked to or associated with the FVIII protein.

### Claim 121 (depends on 120)

The polynucleotide of claim 120, further comprising a polynucleotide chain, which encodes PC5 or PC7.

### Claim 122 (depends on 120)

A vector comprising the polynucleotide of claim 120 and one or more promoter operably linked to the polynucleotide or the set of polynucleotides.

### Claim 123 (depends on 122)

The vector of claim 122, further comprising an additional vector, which comprises a polynucleotide chain encoding PC5 or PC7.

### Claim 124 (depends on 120)

A host cell comprising the polynucleotide of any one of claims 120.

### Claim 125 (depends on 124)

The host cell of claim 124, which is a mammalian cell.

### Claim 126 (depends on 125)

The host cell of claim 125, wherein the mammalian cell is selected from the group consisting of HEK293 cell, CHO cell, and BHK cell.

### Claim 127 (depends on 120)

A pharmaceutical composition comprising the chimeric protein encoded by the polynucleotide of claim 120.

### Claim 128 (depends on 127)

The composition of claim 127, wherein the FVIII protein has extended half-life compared to wild type FVIII protein.

### Claim 129 (depends on 128)

The composition of claim 128, wherein the half-life of the FVIII protein is extended at least about 1.5 times, at least about 2 times, at least about 2.5 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least about 9 times, at least about 10 times, at least about 11 times, or at least about 12 times longer than wild type FVIII.

### Claim 130 (depends on 129)

The composition of claim 129, wherein the half-life of Factor VIII is at least about 17 hours, at least about 18 hours, at least about 19 hours, at least about 20 hours, at least about 21 hours, at least about 22 hours, at least about 23 hours, at least about 24 hours, at least about 25 hours, at least about 26 hours, at least about 27 hours, at least about 28 hours, at least about 29 hours, at least about 30 hours, at least about 31 hours, at least about 32 hours, at least about 33 hours, at least about 34 hours, at least about 35 hours, at least about 36 hours, at least about 48 hours, at least about 60 hours, at least about 72 hours, at least about 84 hours, at least about 96 hours, or at least about 108 hours.

### Claim 131 (depends on 127)

The composition of claim 127, which is administered by a route selected from the group consisting of topical administration, intraocular administration, parenteral administration, intrathecal administration, subdural administration and oral administration.

### Claim 132 (depends on 131)

The composition of claim 131, wherein the parenteral administration is intravenous or subcutaneous administration.

### Claim 133 (depends on 127)

The composition of any one of claims 127, which is used to treat a bleeding disease or condition in a subject in need thereof.

### Claim 134 (depends on 133)

The composition of claim 133, wherein the bleeding disease or condition is selected from the group consisting of a bleeding coagulation disorder, hemarthrosis, muscle bleed, oral bleed, hemorrhage, hemorrhage into muscles, oral hemorrhage, trauma, trauma capitis, gastrointestinal bleeding, intracranial hemorrhage, intra-abdominal hemorrhage, intrathoracic hemorrhage, bone fracture, central nervous system bleeding, bleeding in the retropharyngeal space, bleeding in the retroperitoneal space, bleeding in the illiopsoas sheath and any combinations thereof.

### Claim 135 (depends on 133)

The composition of claim 133, wherein the subject is scheduled to undergo a surgery.

### Claim 136 (depends on 133)

The composition of claim 133, wherein the treatment is prophylactic or on-demand.

### Claim 137 (depends on 120)

A method of preventing or inhibiting binding of a FVIII protein with endogenous VWF comprising adding an effective amount of the chimeric protein encoded by the polynucleotide of claim 120 to a subject in need thereof, wherein the VWF fragment binds to the FVIII protein and thus prevents or inhibits binding of endogenous VWF.

### Claim 138 (depends on 120)

A method of extending or increasing half-life of a FVIII protein, wherein the method comprises adding an effective amount of the chimeric protein encoded by the polynucleotide of claim 120 to a subject in need thereof, wherein the VWF fragment binds to the FVIII protein and thus extends or increases half-life of the FVIII protein.

### Claim 139 (depends on 120)

A method of preventing or inhibiting clearance of a FVIII protein from a cell, wherein the method comprises adding an effective amount of the chimeric protein encoded by the polynucleotide of claim 120 to a cell comprising a FVIII protein or a polynucleotide encoding the FVIII protein, wherein the protein having VWF activity binds to the FVIII protein.

### Claim 140 (depends on 137)

The method of any one of claims 137, wherein the subject is an animal.

### Claim 141 (depends on 140)

The method of claim 140, wherein the animal is a human.

### Claim 142 (depends on 140)

The method of claim 140, wherein the subject is suffering from hemophilia A.

### Claim 143 (depends on 120)

A method of treating a bleeding disease or disorder in a subject in need thereof comprising administering an effective amount of the chimeric protein encoded by the polynucleotide of claim 120, wherein the bleeding disease or disorder is selected from the group consisting of a bleeding coagulation disorder, hemarthrosis, muscle bleed, oral bleed, hemorrhage, hemorrhage into muscles, oral hemorrhage, trauma, trauma capitis, gastrointestinal bleeding, intracranial hemorrhage, intra-abdominal hemorrhage, intrathoracic hemorrhage, bone fracture, central nervous system bleeding, bleeding in the retropharyngeal space, bleeding in the retroperitoneal space, and bleeding in the illiopsoas sheath.

### Claim 144 (depends on 143)

The method of claim 143, wherein the treatment is prophylactic or on-demand.

### Claim 145 (depends on 143)

The method of claim 143, wherein the effective amount is 0.1 μg/kg to 500 mg/kg.

### Claim 146 (depends on 143)

The method of claim 143, wherein the chimeric protein is administered by a route selected from the group consisting of topical administration, intraocular administration, parenteral administration, intrathecal administration, subdural administration and oral administration.

### Claim 147 (depends on 146)

The method of claim 146, wherein the parenteral administration is selected from the group consisting of intravenous administration, subcutaneous administration, intramuscular administration, and intradermal administration,

### Claim 148 (depends on 120)

A method of making a chimeric protein, comprising transfecting one or more host cell with the polynucleotide of claim 120 and expressing the chimeric protein in the host cell.

### Claim at position 31

149-163. (canceled)

### Claim 164 (depends on 124)

The host cell of claim 124, wherein the PC5 or PC7 cleaves the D1D2 domains of VWF.

### Claim 165

(canceled)