Modified Human Four Helical Bundle Polypeptides and Their Uses

• US 20080097083A1
• Filed: 10/18/2007
• Published: 04/24/2008
• Est. Priority Date: 02/02/2004
• Status: Active Grant

• Current Assignee: Ambrx, Inc. (Johnson & Johnson)
• Original Assignee: Ambrx, Inc. (Johnson & Johnson)
• Inventors: LITZINGER, David, CHO, Ho Sung, HAYS, Anna-Maria, DIMARCHI, Richard, SIM, Bee-Cheng, WILSON, Troy, DANIEL, Thomas
• Granted Patent: US 8,907,064 B2
• Time in Patent Office: Days
• US Class Current: 530/350
• 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. A four helical bundle (4HB) polypeptide comprising one or more non-naturally encoded amino acids wherein said non-naturally encoded amino acids are substituted at one or more positions in the A helix, B helix, C helix, or D helix.

2. The 4HB polypeptide of claim 1, wherein the 4HB polypeptide further comprises one or more amino acid substitutions made within the N-terminus, C-terminus, 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 comprises one or more 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 comprises one or more 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 comprises one or more amino acid substitution, addition or deletion that increases the expression of the 4HB polypeptide in a recombinant host cell or synthesized in vitro.

6. The 4HB polypeptide of claim 1, wherein the 4HB polypeptide comprises one or more amino acid substitution, addition or deletion that increases protease resistance of the 4HB polypeptide.

7. The 4HB polypeptide of claim 1, wherein the non-naturally encoded amino acid is selectively reactive toward 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, an aminooxy group, a hydrazine group, a hydrazide group, a semicarbazide group, an azide group, or an alkyne 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, selected from the group consisting of granulocyte cell stimulating factor (G-CSF), erythropoietin, interferon, and growth hormone.

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

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

13. 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.

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

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

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

17. 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.

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

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

20-83. -83. (canceled)

84. A polypeptide from the growth hormone supergene family comprising one or more non-naturally encoded amino acids wherein said non-naturally encoded amino acids are substituted at one or more positions in the E helix.

85. The polypeptide of claim 84, wherein the polypeptide further comprises one or more amino acid substitutions made within the N-terminus, C-terminus, A-B loop, B-C loop, C-D loop, or D-E loop.