# US 5,705,479 A · Site-directed mutagenesis modified glycoprotein hormones and methods of use

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- Number: US 5,705,479 A
- Document ID: US5705479A
- Status: Granted
- Kind: A
- Application number: 08/442,963
- Filed: 1995-05-17
- Granted: 1998-01-06
- Earliest claimed: 1987-12-21
- Assignees: Applied Research Systems ARS Holding N.V.
- Inventors: Scott C. Chappel; Edward G. Bernstine
- Web page: https://patentgrep.com/patent/US5705479A

## CPC

- C07K 14/59: Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g.hCG [human chorionic gonadotropin]; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
- 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

Described are new glycoprotein hormones capable of competing with natural hormones for the normal receptor binding sites but substantially incapable of effecting post receptor activities. The glycoprotein hormones of the present invention have had specific (rather than all) oligosaccharide chains removed so as to effectively diminish biologic activity while not significantly reducing plasma half-life, thus improving the molecules effectiveness as an antagonist compared with conventionally-produced molecules. The preferred glycoprotein hormones are ideally obtained by site-directed mutagenesis to selectively deglycosylate the protein. Also described are therapeutic treatments comprising the administration of the recombinant glycoprotein hormones of the present invention as hormone antagonists.

## Claims

### Claim 1

A method for treating the overproduction of a dimeric glycoprotein hormone selected from the group consisting of luteinizing hormone, follicle stimulating hormone, thyroid stimulating hormone, and human chorionic gonadotropin, comprising administering a therapeutically effective dose of a corresponding hormone which has at least one N-linked oligosaccharide chain entirely removed so as to be capable of competing with the corresponding natural hormone for binding to a natural receptor molecule but incapable of stimulating post receptor biological activity, and having a plasma half-life approximately equal to that of the corresponding natural hormone.

### Claim 2 (depends on 1)

A method in accordance with claim 1, wherein said glycoprotein hormone is human chorionic gonadotropin.

### Claim 3 (depends on 1)

A method in accordance with claim 1, wherein said N-linked oligosaccharide chain which has been removed from said hormone is or are removed from at least one of the two glycosylation sites on the .alpha. subunit thereof without removal of any oligosaccharide chains from the .beta. subunit thereof.

### Claim 4 (depends on 3)

A method in accordance with claim 3, wherein all of the N-linked oligosaccharide chains have been removed from said .alpha. subunit.

### Claim 5 (depends on 3)

A method in accordance with claim 3, wherein the N-linked oligosaccharide chains have been removed from only one of the two glycosylation sites on said .alpha. subunit.

### Claim 6 (depends on 5)

A method in accordance with claim 5, wherein said glycosylation site from which the N-linked oligosaccharide chains have been removed is at the 52 amino acid position.