# US 5,707,832 A · Process for the preparation of human factor VIII and analogs of factor VIII

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

- Number: US 5,707,832 A
- Document ID: US5707832A
- Status: Granted
- Kind: A
- Application number: 08/384,774
- Filed: 1995-02-07
- Granted: 1998-01-13
- Earliest claimed: 1990-02-05
- Assignees: TM Innovation
- Inventors: Gerard Mignot; Nicolas Bihoreau; Philippe Adamowicz
- Web page: https://patentgrep.com/patent/US5707832A

## CPC

- C07K 14/755: Factors VIII {, e.g. factor VIII C (AHF), factor VIII Ag (VWF)}
- 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: 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)

## Abstract

The present invention relates to a process for the preparation of factor VIII or an analog of factor VIII by culture, in a culture medium, of cells which produce said factor VIII or analog of factor VIII and the separation of factor VIII or its analog, wherein the culture medium contains at least one derivative of a polycationic and/or polyanionic polymer. It also relates to factor VIII or analog of factor VIII and to the complex of factor VIII or analog of factor VIII with a derivative of a polycationic and/or polyanionic polymer obtained by using this process.

## Claims

### Claim 1

In a process for the preparation and separation of factor VIII or an analog of factor VIII, the process comprising culturing mammalian eukaryotic cells which produce and secrete said factor VIII or analog of factor VIII, said mammalian eukaryotic cells having incorporated an expression vector containing a gene encoding for said factor VIII or analog of factor VIII, proviso they are not chosen among: Vero, Hela, WI38, BIIK, CHO, COS-7 and MDCK, said mammalian cells expressing the molecule of factor VIII or analog of factor VIII in a continuous mariner, and then separating said factor VIII or analog of factor VIII wherein the improvement comprises culturing the mammalian cells in a serum free culture medium and free of Von Willebrand factor, supplemented with at least one derivative which is a sulfated polysaccharide of molecular weight between 5,000 and 700,000 and a degree of sulfation of 0.5% to 18% by weight of sufur, in an amount sufficient to provide an enhancement of factor VIII or analog of factor VIII production compared with culturing the mammalian cells in a medium without said sulfated polysaccharide.

### Claim 2 (depends on 1)

The process of claim 1 wherein the sulfated dextran preferably has a degree of sulfation of 10% to 18%.

### Claim 3 (depends on 1)

The process of claim 1, wherein the sulfated polysaccharide is chosen from the group consisting of heparin, sulfated dextran and hydroxy ethylstarch.

### Claim 4 (depends on 1)

The process claim 1, wherein the analog of factor VIII which is prepared is factor VIII delta 2.

### Claim 5

In a process for the preparation and separation of factor VIII or an analog of factor VIII, the process comprising culturing mammalian eukaryotic cells which produce and secrete said factor VIII or analog of factor VIII, said mammalian eukaryotic cells having incorporated an expression vector containing a gene encoding for said factor VIII or analog of factor VIII said cells being chosen among recombinant Veto, Hela, WI38, BHK, CHO, COS-7 and MDCK cells which express the molecule of factor VIII or analog of factor VIII in a continuous manner, and then separating said factor VIII or analog of factor VIII, wherein the improvement comprises culturing the mammalian cells in a serum free culture medium and free of Von Willebrand factor, supplemented with at least one derivative which is a sulfated polysaccharide of molecular weight between 5,000 and 700,000 and a degree of sulfation of 65% to 18% by weight of sulfur, in an amount sufficient to provide an enhancement of factor VIII or analog of factor VIII production compared with culturing the mammalian cells in a medium without said sulfated polysaccharide proviso that said sulfated polysaccharide is not chosen among heparin, sulfated dextran and hydroxyethylstarch sulfate of molecular weight between 5000 and 700,000 and a degree of sulfation of 0.5% to 18% by weight of sulfur.

### Claim 6 (depends on 5)

The process of claim 5, wherein the analog of factor VIII which is prepared is factor VIII delta 2.

### Claim 7 (depends on 5)

The process of claim 5, wherein the cells in culture are recombinant CHO cells which express the molecule of factor VIII or analog of factor VIII in a continuous manner.

### Claim 8

A process for the preparation and separation of factor VIII or an analog of factor VIII, the process comprising culturing mammalian eukaryotic cells which produce and secrete said factor VIII or analog of factor VIII, said mammalian eukaryotic cells having incorporated an expression vector containing a gene encoding for said factor VIII or analogue of factor VIII, proviso though they are not chosen among: Vero, Hela, WI38, BIIK, CHO, COS-7, MDCK, said mammalian cells expressing the molecule factor VIII or analog of factor VIII in a continuous manner, and then separating said factor VIII or analog of factor VIII wherein the improvement comprises culturing the mammalian cells in a serum free culture median and free of Von Willebrand factor, supplemented with a sulfated polysaccharide selected from the group consisting of heparin, sulfated dextran, and hydroxyethyl starch, in an amount sufficient to provide an enhancement of factor VIII or analog of factor VIII production compared with culturing the mammalian cells in a medium without said supplement.