Modified human four helical bundle polypeptides and their uses

• US 8,907,064 B2
• Filed: 10/18/2007
• Issued: 12/09/2014
• Est. Priority Date: 02/02/2004
• Status: Active Grant

• Current Assignee: Ambrx, Inc. (Johnson & Johnson)
• Original Assignee: Ambrx, Inc. (Johnson & Johnson)
• Inventors: Cho, Ho Sung, Daniel, Thomas O., DiMarchi, Richard D., Hays, Anna-Maria, Wilson, Troy E., Sim, Bee-Cheng, Litzinger, David C.
• Primary Examiner(s): Shafer, Shulamith H
• Application Number:
  US11/874,713
• Publication Number:
  US 20080097083A1
• Time in Patent Office: 2,609 Days
• Field of Search: None
• US Class Current: 530/351
• CPC Class Codes: A61K 38/21 Interferons [IFN]
• CPC Class Codes: A61K 38/27 Growth hormone [GH], i.e. somatotropin
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 13/08 of the prostate
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/00 Drugs for disorders of the endocrine system
• CPC Class Codes: A61P 5/06 of the anterior pituitary hormones, e.g. TSH, ACTH, FSH, LH, PRL, GH
• CPC Class Codes: A61P 5/10 of the posterior pituitary hormones, e.g. oxytocin, ADH
• CPC Class Codes: A61P 5/12 : for decreasing, blocking or antagonising the activity of the posterior pituitary hormones
• CPC Class Codes: A61P 7/06 : Antianaemics
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07H 21/04 : with deoxyribosyl as saccharide radical
• CPC Class Codes: C07K 14/475 : Growth factors; Growth regulators
• CPC Class Codes: C07K 14/52 : Cytokines; Lymphokines; Interferons
• 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
• CPC Class Codes: C08G 65/33396 : having oxygen in addition to nitrogen
• CPC Class Codes: G01N 2500/00 : Screening for compounds of potential therapeutic value
• CPC Class Codes: G01N 33/6875 : Nucleoproteins

Abstract

Modified human four helical bundle (4HB) polypeptides and uses thereof are provided.

Claims

1. An isolated four helical bundle (4HB) polypeptide chosen from the group consisting of human growth hormone, interferon, erythropoietin, and granulocyte cell stimulating factor, comprising a ribosomally incorporated para-acetylphenylalanine wherein said para-acetylphenylalanine is substituted at the C-terminus, wherein the 4HB polypeptide with a non-naturally encoded amino acid has at least a 2-fold increased serum half-life over the wild type 4HB polypeptide.

2. The 4HB polypeptide of claim 1, wherein the 4HB polypeptide further comprises up to five amino acid substitutions made within the A Helix, B Helix, C Helix, D Helix, A-B loop, B-C loop, C-D loop, or, when present, the E Helix or D-E loop.

3. The 4HB polypeptide of claim 1, wherein the 4HB polypeptide further comprises up to five amino acid substitutions, additions or deletions that modulate affinity of the 4HB polypeptide for a 4HB receptor.

4. The 4HB polypeptide of claim 1, wherein the 4HB polypeptide further comprises up to five amino acid substitutions, additions or deletions that increase the stability or solubility of the 4HB polypeptide.

5. The 4HB polypeptide of claim 1, wherein the 4HB polypeptide further comprises up to five amino acid substitution(s), addition(s) or deletion(s) that increase the expression of the 4HB polypeptide in a recombinant host cell or increases synthesis of the 4HB polypeptide in vitro.

6. The 4HB polypeptide of claim 1, wherein the 4HB polypeptide further comprises up to five amino acid substitution(s), addition(s) or deletion(s) that increases protease resistance of the 4HB polypeptide.

7. The 4HB polypeptide of claim 1, wherein the non-naturally encoded amino acid includes a side chain functional group that reacts selectively with a linker, polymer, or biologically active molecule.

8. The 4HB polypeptide of claim 1, wherein the non-naturally encoded amino acid comprises a carbonyl group.

9. The 4HB polypeptide of claim 1, wherein the 4HB polypeptide comprises one or more post-translational modifications.

10. The 4HB polypeptide of claim 1, wherein the 4HB polypeptide is linked to a linker, polymer, or biologically active molecule.

11. The 4HB polypeptide of claim 10, wherein the 4HB polypeptide is linked to a water soluble polymer.

12. The 4HB polypeptide of claim 1, wherein the 4HB polypeptide is linked to a multifunctional polymer, bifunctional linker, or at least one additional 4HB polypeptide.

13. The 4HB polypeptide of claim 12, wherein the bifunctional linker or bifunctional polymer is linked to a second polypeptide.

14. The 4HB polypeptide of claim 13, wherein the second polypeptide is a 4HB polypeptide.

15. The 4HB polypeptide of claim 11, wherein the water soluble polymer comprises a poly (ethylene glycol) moiety.

16. The 4HB polypeptide of claim 11, wherein said water soluble polymer is linked to a non-naturally encoded amino acid present in said 4HB polypeptide.

17. The 4HB polypeptide of claim 1, wherein the 4HB polypeptide further comprises one or more free amide or free sulfhydryl groups.

18. The 4HB polypeptide of claim 17, wherein the free amide or free sulfhydryl groups are present in naturally occurring amino acids.