Biosynthetic polypeptides utilizing non-naturally encoded amino acids

• US 7,638,299 B2
• Filed: 07/21/2005
• Issued: 12/29/2009
• Est. Priority Date: 07/21/2004
• Status: Expired due to Fees

• Current Assignee: Ambrx, Inc. (Johnson & Johnson)
• Original Assignee: Ambrx, Inc. (Johnson & Johnson)
• Inventors: Keefe, William M., Cho, Ho Sung, Litzinger, David C., Hays, Anna-Maria, Daniel, Thomas O., Wilson, Troy E., Mariani, Roberto, Kimmel, Bruce E.
• Primary Examiner(s): Wang; Andrew
• Assistant Examiner(s): Gebreyesus; Kagnew H
• Application Number:
  US11/187,687
• Publication Number:
  US 20060019347A1
• Time in Patent Office: 1,622 Days
• Field of Search: None
• US Class Current: 435/69.1
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 1/14 Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• 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 3/02 Nutrients, e.g. vitamins, minerals
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 5/00 Drugs for disorders of the endocrine system
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61P 9/12 Antihypertensives
• CPC Class Codes: C07K 14/005 from viruses
• 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 14/57563 Vasoactive intestinal peptide [VIP]; Related peptides
• CPC Class Codes: C07K 14/605 Glucagons
• CPC Class Codes: C07K 2319/02 containing a signal sequence
• CPC Class Codes: C07K 2319/20 containing a tag with affinity for a non-protein ligand
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins
• CPC Class Codes: C12N 2740/16122 : New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Abstract

Modified biosynthetic polypeptide molecules, methods for manufacturing, and uses thereof are provided.

Claims

1. A method for making a biosynthetic polypeptide (BSP) less than 100 amino acids in length comprising two or more non-naturally encoded amino acids wherein said BSP is a (glucagon like polypeptide 1 (GLP-1) comprising;

expressing an mRNA encoding said GLP-1 peptide in a translation system, wherein said mRNA comprises two or more selector codons, and said translation system further comprises an orthogonal aminoacyl tRNA synthetase (O-RS) and an orthogonal tRNA (O-tRNA) wherein said O-RS preferentially aminoacylates said O-tRNA with said non-naturally encoded amino acids, and said O-tRNA recognizes the two or more selector codons on the mRNA and wherein at least two of the non-naturally encoded amino acids are linked together.

2. The method of claim 1, wherein the BSP further comprises at least one poly(ethylene glycol) moiety.

3. The method of claim 1, wherein the non-naturally encoded amino acids in the BSP are substituted at a position selected from the group consisting of residues at, H7, A8, E9, V16, S17, S18, Y19, L20, E21, G22, Q23, A24, A25, K26, E27, F28, I29, A30, W31, L32, V33, K34, G35, R36, G37, and any combination thereof wherein position 7-37 of GLP-1 correspond to position 1-31 of SEQ ID NO:

2.

4. The method of claim 1, wherein one or more of the non-naturally encoded amino acids in the BSP has a functional group determined to cleave a peptide bond under selected conditions selected from a group consisting of a photo-activated functional group, a pH activated functional group, a temperature activated functional group, a functional group that requires a catalyst, a functional group that is recognized by a protease, and a functional group that is recognized by an enzyme.

5. The method of claim 4, wherein the selected conditions of the BSP are selected from a group consisting of exposure to photon energy, increased temperature, decreased temperature, increased pH, decreased pH, exposure to sub-atomic particles, addition of a catalyst, incubation with an enzyme, and contact with another chemical functional group.

6. The method of claim 2, wherein the BSP comprising a poly(ethylene glycol) moiety has a molecular weight of between about 0.1 kDa and about 100 kDa.

7. The method of claim 2, wherein the BSP comprising a poly(ethylene glycol) moiety is a branched or multiarmed polymer.

8. The method of claim 7, wherein the BSP comprising a poly(ethylene glycol) moiety has a molecular weight of between about 1 kDa and about 100 kDa.

9. The method of claim 1, wherein the BSP comprising a polyalkylene glycol has a molecular weight of between about 0.1 kDa and about 50 kDa.

10. The method of claim 1, wherein the BSP further comprises at least one polyalkylene glycol polymer.

11. The method of claim 3, wherein the non-naturally encoded amino acids in the BSP further comprises an addition at a position selected from the group consisting of a position before the amino acid residue at position 7, and an addition at position 38, and any combination thereof wherein position 7-37 of GLP-1 correspond to position 1-31 of SEQ ID NO:

2.