Non-natural amino acid replication-dependent microorganisms and vaccines

• US 9,121,025 B2
• Filed: 07/07/2013
• Issued: 09/01/2015
• Est. Priority Date: 09/26/2008
• Status: Active Grant

• Current Assignee: Ambrx, Inc. (Johnson & Johnson)
• Original Assignee: Ambrx, Inc. (Johnson & Johnson)
• Inventors: Tian, Feng, Hehli, Brad
• Primary Examiner(s): Nickol, Gary
• Assistant Examiner(s): SHAHNAN SHAH, KHATOL S
• Application Number:
  US13/936,201
• Publication Number:
  US 20130280301A1
• Time in Patent Office: 786 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 39/0258 Escherichia
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: C12N 1/36 Adaptation or attenuation of cells
• CPC Class Codes: C12N 15/70 Vectors or expression systems specially adapted for E. coli
• CPC Class Codes: C12N 9/93 Ligases (6)

Abstract

Compositions and methods of producing vaccines, including methods wherein whole organism vaccines are provided with limited replication abilities, thereby increasing vaccine safety and efficacy, through the use of non-natural, unnatural, or non-naturally encoded amino acids.

Claims

1. A non-human animal vaccine, comprising an Escherichia coli (E. coli) cell and an adjuvant, wherein the E. coli cell comprises a non-natural amino acid, wherein the non-natural amino acid has been site-specifically incorporated into an essential gene product for replication, wherein the essential gene product is encoded by a gene selected from the group consisting of ileS, lspA, ispH, ftsL, ftsI, murE, murD, ftsW, murG, murC, ftsQ, ftsA, ftsZ, secA, map, rpsB, pyrH, frr, uppS, cdsA, fabZ, lpxA, lpxB, dnaE, accA, proS, nusB, dxs, ispA, dnaX, adk, cysS, mrdB, mrdA, holA, leuS, glnS, serS, msbA, kdsB, asnS, fabA, rne, fabD, fabG, tmk, holB, lolC, lolD, lolE, trmU, prsA, topA, fabI, pheS, rplT, infC, nadE, aspS, argS, rplY, nrdA, nrdB, fabB, gltX, ligA, zipA, yfgE, hisS, acpS, rplS, trmD, rpsP, ffh, grpE, plsC, parC, parE, ribB, rpoD, rpsO, yhbZ, rpsI, rplM, def, rpsM, secY, rplO, rpsE, rplR, rplF, rpsN, rplX, rplN, rpsQ, rpsS, rplB, rplW, rplD, rplC, rpsJ, fusA, rpsL, trpS, ftsY, glyS, kdtA, coaD, dut, gmk, dnaA, glmU, engB, murI, rplJ, rplL, rpoB, rpoC, dnaB, groS, groL, efp, rpsf, ppa, valS, dnaC, and dnaT;

and

wherein said E. coil cell is capable of replication only in the presence of said non-natural amino acid, and has limited or no replication capability in the absence of said non-natural amino acid.

2. The non-human animal vaccine of claim 1, wherein more than one non-natural amino acid is site-specifically incorporated into the essential gene product for replication.

3. The non-human animal vaccine of claim 1, wherein more than one non-natural amino acid is incorporated into more than one essential gene product for replication.

4. The non-human animal vaccine of claim 1, wherein the non-natural amino acid is selected from the group consisting of:

para-acetylphenykilanine, p-nitrophenylalanine, p-sulfotyrosine, p-carboxyphenylalanine, an o-nitrophenylalanine, an m-nitrophenylalanine, a p-boronyl phenylalanine, an o-boronylphe,nylalanine, an m-boronylphenylalanine, a p-aminophenylalanine, an o-aminophenylalanine, an m-aminophenylalanine, a p-acylphenylalanine, an o-acylphenylalanine, an m-acylphenylalanine, a p-OMe phenylalanine, an o-OMe phenylalanine, an m-OMe phenylalanine, a p-sulfophenylalanine, an o-sulfophenylalanine, an m-sulfophenylalanine, a 5-nitro His, a 3-nitro Tyr, a 2-nitro Tyr, a nitro substituted Leu, a nitro substituted His, a nitro substituted De, a nitro substituted Trp, a 2-nitro Trp, a 4-nitro Trp, a 5-nitro Trp, a 6-nitro Trp, a 7-nitro Trp, 3-aminotyrosine, 2-aminotyrosine, O-sulfotyrosine, 2-sulfboxyphenylalanine, 3-sulfooxyphenylalanine, o-earboxyphenylalanine, m-carboxyphenylalanine, p-acetyl-L-phenylalanine, a p-propargyl-phenylalanine, O-methyl-L-tyrosine, an L-3-(2-naphthyl)aelanine, a 3-methyl-phenylalanine, an O-4-allyl-L-tyrosine, a 4-propyl-L-tyrosine, a tri-O-acetyl-GleNAcβ

-serine, an L-Dopa, a fluorinated phenylalanine, an isopropyl-L-phenylaianine, a p-azido-L-phenylalanine, a p-acyl-L-phenylaianine, a p-benzoyl-L-phenylalanine, an L-phosphoserine, a phosphonoserine, a phosphonotyrosine, a p-iodo-phenylalanine, a p-bromophenylalanine, a p-amino-L-phenylalanine, an isopropyl-L-phenylalanine, and a p-propargyloxy-phenyialanine.

5. The non-human animal vaccine according to claim 4, wherein the non-natural amino acid is para-acetylphenylalanine.

6. The non-human animal vaccine according to claim 1, wherein the E. coil cell comprises a non-natural amino acid incorporated into the amino acid sequence of essential gene product methionine amino peptidase (map).

7. The non-human animal vaccine according to claim 1, wherein the E. coil cell comprises a para-acetylphenylalanine incorporated into the amino acid sequence of essential gene product methionine amino peptidase (map).

8. A method of inducing an immune response or reducing the risk of an E. coil infection, said method comprising administering to a non-human animal an effective amount of the nonhuman animal vaccine of claim 1.

9. The method of claim 8, wherein said non-human animal is selected from the group consisting of:

cow, pig, goat, sheep, horse, and dog.

10. A method of producing antibodies against E. coil, said method comprising administering to a non-human animal an effective amount of the non-human animal vaccine of claim 1 to produce antibodies against E. coli.