Protease resistant interferon beta mutants

• US 7,998,469 B2
• Filed: 02/13/2007
• Issued: 08/16/2011
• Est. Priority Date: 09/09/2002
• Status: Expired due to Fees

• Current Assignee: HanAll Biopharma Co. Ltd.
• Original Assignee: HanAll Biopharma Co. Ltd.
• Inventors: Gantier, Rene, Guyon, Thierry, Drittanti, Lila, Vega, Manuel
• Primary Examiner(s): STOICA, ELLY GERALD
• Application Number:
  US11/706,088
• Publication Number:
  US 20070172459A1
• Time in Patent Office: 1,645 Days
• Field of Search: None
• US Class Current: 424/85.6
• CPC Class Codes: 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
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/08 Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07K 14/475 Growth factors; Growth regulators
• CPC Class Codes: C07K 14/505 Erythropoietin [EPO]
• CPC Class Codes: C07K 14/52 Cytokines; Lymphokines; Interferons
• CPC Class Codes: C07K 14/53 Colony-stimulating factor [CSF]
• CPC Class Codes: C07K 14/535 Granulocyte CSF; Granulocyte-macrophage CSF
• CPC Class Codes: C07K 14/54 Interleukins [IL]
• CPC Class Codes: C07K 14/555 : Interferons [IFN]
• CPC Class Codes: C07K 14/575 : Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin C07K14/665, e.g. corticotropin C07K14/695
• CPC Class Codes: C07K 14/61 : Growth hormone [GH], i.e. somatotropin

Abstract

Compositions of modified cytokines and uses thereof generated using processes and systems for the high throughput directed evolution of peptides and proteins, particularly cytokines that act in complex biological settings, are provided. Also provided are modified cytokines formulated for oral delivery and uses thereof to treat diseases and conditions mediated by cytokines.

Claims

1. A modified interferon beta (IFN beta) cytokine, wherein said modified IFN beta cytokine differs from an unmodified IFN beta cytokine by two amino acid substitutions

wherein said unmodified IFN beta cytokine comprises the amino acid sequence of SEQ ID NO:

196, or comprises a polypeptide with anti-proliferative activity having an amino acid sequence at least 95% identical to SEQ ID NO;

196

said two amino acid substitutions being either;

(a) substitution of the 5th and 108th positions in SEQ ID NO;

196, or of the corresponding positions in the unmodified IFN beta cytokine comprising a polypeptide with anti-proliferative activity having an amino acid sequence at least 95% identical to SEQ ID NO;

196, with aspartic acid and serine, respectively;

or

(b) substitution of the 6th and 108th positions in SEQ ID NO;

196, or of the corresponding positions in the unmodified IFN beta cytokine comprising a polypeptide with anti-proliferative activity having an amino acid sequence at least 95% identical to SEQ ID NO;

196, with glutamine and serine, respectively,

and wherein said two amino acid substitutions confer increased resistance to proteolysis over the unmodified IFN beta cytokine.

2. The modified IFN beta cytokine of claim 1, wherein the unmodified IFN beta cytokine comprises the amino acid sequence of SEQ ID NO:

196 or 197.

3. A pharmaceutical composition comprising the modified IFN beta cytokine of claim 1.

4. The pharmaceutical composition of claim 3, further comprising a pharmaceutically acceptable carrier or excipient selected from the group consisting of a binding agent, a filler, a lubricant, a disintegrant, and a wetting agent.

5. The pharmaceutical composition of claim 3, further comprising a pharmaceutically acceptable additive.

6. The pharmaceutical composition of claim 5, wherein the pharmaceutically acceptable additive is selected from the group consisting of a suspending agent, an emulsifying agent, a non-aqueous vehicle, and a preservative.

7. The pharmaceutical composition of claim 3, wherein the composition is in the form of a liquid, a solution, a suspension, an aerosol, a tablet, a lozenge or a capsule.

8. The pharmaceutical composition of claim 7, wherein the composition is in the form of a tablet or a capsule.

9. The pharmaceutical composition of claim 3, formulated for oral, parenteral, intravenous, intradermal, subcutaneous, buccal, inhalation, intramuscular, rectal or topical administration.

10. The pharmaceutical composition of claim 9, formulated for oral administration.

11. The pharmaceutical composition of claim 3, wherein the pharmaceutical composition is formulated for controlled-release of the modified IFN beta cytokine.