D-amino acid derivative-modified peptidoglycan and methods of use thereof

Patent metadata

Field	Value
Publication number	US-10016498-B2
Application number	US-201715704977-A
Country	US
Kind code	B2
Filing date	Sep 14, 2017
Priority date	Nov 30, 2012
Publication date	Jul 10, 2018
Grant date	Jul 10, 2018

Abstract

The present disclosure provides modified bacteria and modified peptidoglycan comprising modified D-amino acids; compositions comprising the modified bacteria or peptidoglycan; and methods of using the modified bacteria or peptidoglycan. The modified D-amino acids include a bioorthogonal functional group such as an azide, an alkyne or a norbornene group. Also provided are modified peptidoglycans conjugated to a molecule of interest via a linker.

First claim

What is claimed is:

1. A method of identifying an agent that inhibits peptidoglycan synthesis, the method comprising:
a) contacting a bacterial cell in vitro with a test agent; and
b) determining the effect, if any, of the test agent on incorporation of a modified D-amino acid into peptidoglycan in the bacterial cell, wherein the modified D-amino acid is an azide-modified, an alkyne-modified, or a norbornene-modified D-amino acid;
wherein a test agent that inhibits incorporation of the modified D-amino acid into peptidoglycan in the bacterial cell is considered a candidate agent for inhibiting peptidoglycan synthesis.

2. The method of claim 1 , wherein the bacterial cell is present intracellularly in a eukaryotic host cell.

3. The method of claim 1 , wherein the eukaryotic host cell is a macrophage.

4. The method of claim 1 , wherein incorporation of the modified D-amino acid into the peptidoglycan is determined by contacting the cell with a reagent selected from a detectably labeled azide-containing reagent, a detectably labeled alkynyl reagent and a detectably labeled norbornene-reactive reagent.

5. The method of claim 4 , wherein the detectable label is a fluorescent label.

6. The method of claim 1 , wherein the modified D-amino acid is an azide-modified D-amino acid.

7. The method of claim 1 , wherein the modified D-amino acid is an alkyne-modified D-amino acid.

8. The method of claim 1 , wherein the modified D-amino acid is a norbornene-modified D-amino acid.

9. The method of claim 6 , wherein incorporation of the modified D-amino acid into the peptidoglycan is determined by contacting the cell with a detectably labeled alkynyl reagent.

10. The method of claim 7 , wherein incorporation of the modified D-amino acid into the peptidoglycan is determined by contacting the cell with a detectably labeled azide-containing reagent.

11. The method of claim 8 , wherein incorporation of the modified D-amino acid into the peptidoglycan is determined by contacting the cell with a detectably labeled norbornene-reactive reagent.

12. The method of claim 4 , wherein the detectable label is a radioactive isotope.

13. The method of claim 4 , wherein the detectable label is biotin.

14. The method of claim 4 , wherein the detectable label is a peptide that can be detected by antibody binding.

Assignees

• Univ California

Inventors

• Bertozzi Carolyn R
• Sloan Siegrist Palmore Mary
• Jewett John C
• Gordon Chelsea G
• Shieh Peyton

Classifications

• A61K39/385 Primary

  Haptens or antigens, bound to carriers · CPC title

• C12Q1/025 Primary

  for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics (antimicrobial activity C12Q1/18) · CPC title

• G01N2333/195

  from bacteria · CPC title

• A61K2039/6068

  Other bacterial proteins, e.g. OMP · CPC title

• A61K47/646

  the entire peptide or protein drug conjugate elicits an immune response, e.g. conjugate vaccines · CPC title