# US 10,370,431 B2 · Optimized factor VIII gene

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 10,370,431 B2
- Document ID: US10370431B2
- Status: Granted
- Kind: B2
- Application number: 14/767,425
- Filed: 2014-02-14
- Published: 2015-12-17
- Granted: 2019-08-06
- Earliest claimed: 2013-02-15
- Assignees: Bioverativ Therapeutics Inc.
- Applicants: Bioverativ Therapeutics Inc.
- Inventors: Siyuan Tan; Robert T. Peters
- Web page: https://patentgrep.com/patent/US10370431B2

## CPC

- C07K 14/755: Factors VIII {, e.g. factor VIII C (AHF), factor VIII Ag (VWF)}
- C12N 9/6424: {Serine endopeptidases (3.4.21)}
- 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: 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)
- 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 codon optimized Factor VIII sequences, vectors and host cells comprising codon optimized Factor VIII sequences, polypeptides encoded by codon optimized Factor VIII sequences, and methods of producing such polypeptides.

## Claims

### Claim 1

An isolated nucleic acid molecule comprising a nucleotide sequence having at least 96% sequence identity to SEQ ID NO:2, wherein the nucleotide sequence encodes a polypeptide with Factor VIII activity.

### Claim 2 (depends on 1)

The isolated nucleic acid molecule of claim 1, wherein the nucleotide sequence has one or more of the following characteristics:
a. the nucleotide sequence contains fewer MARS/ARS sequences (SEQ ID NO:5 and SEQ ID NO:6) relative to SEQ ID NO:3;
b. the nucleotide sequence does not contain the splice site GGTGAT (SEQ ID NO:7);
c. the nucleotide sequence contains fewer destabilizing elements (SEQ ID NO:8 and SEQ ID NO:9) relative to SEQ ID NO:3;
d. the nucleotide sequence does not contain a poly-T sequence (SEQ ID NO: 10); and
e. the nucleotide sequence does not contain a poly-A sequence (SEQ ID NO:11).

### Claim 3 (depends on 2)

The isolated nucleic acid molecule of claim 2, wherein the human codon adaptation index is about 0.88.

### Claim 4 (depends on 2)

The isolated nucleic acid molecule of claim 2, wherein the percentage of G/C nucleotides is about 51%.

### Claim 5 (depends on 2)

The isolated nucleic acid molecule of claim 2, wherein the nucleotide sequence contains at most one MARS/ARS sequence.

### Claim 6 (depends on 2)

The isolated nucleic acid molecule of claim 2, wherein the nucleotide sequence contains at most 4 destabilizing elements.

### Claim 7 (depends on 1)

The isolated nucleic acid molecule of claim 1, which further comprises a heterologous nucleotide sequence.

### Claim 8 (depends on 7)

The isolated nucleic acid molecule of claim 7, wherein the heterologous nucleotide sequence encodes a heterologous amino acid sequence that is a half-life extender.

### Claim 9 (depends on 8)

The isolated nucleic acid molecule of claim 8, wherein the heterologous amino acid sequence is an Fc region.

### Claim 10 (depends on 8)

The isolated nucleic acid molecule of claim 8, wherein the heterologous amino acid sequence is linked to the N-terminus or the C-terminus of the amino acid sequence encoded by the nucleotide sequence or inserted between two amino acids in the amino acid sequence encoded by the nucleotide sequence.

### Claim 11 (depends on 1)

The isolated nucleic acid molecule of claim 1, operably linked to at least one transcription control sequence.

### Claim 12 (depends on 1)

A vector comprising the nucleic acid molecule of claim 1.

### Claim 13 (depends on 1)

A host cell comprising the nucleic acid molecule of claim 1.

### Claim 14 (depends on 13)

A method of producing a polypeptide with Factor VIII activity, comprising culturing the host cell of claim 13 under conditions whereby a polypeptide with Factor VIII activity is produced; and, recovering the polypeptide with Factor VIII activity.

### Claim 15 (depends on 12)

The vector of claim 12, wherein the vector is suitable for gene therapy.

### Claim 16 (depends on 15)

The vector of claim 15, wherein the vector is a lentiviral or adeno associated virus (AAV) vector.