Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Methods for treatment of bacterial infections

a technology of bacterial infections and dtranspeptidases, which is applied in the direction of antibacterial agents, biocide, heterocyclic compound active ingredients, etc., can solve the problems of reducing our ability to respond effectively to this threat, reducing the morphology of the colony, and reducing the virulence of the infection

Inactive Publication Date: 2012-05-10
INST NAT DE LA SANTE & DE LA RECHERCHE MEDICALE (INSERM) +1
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0008]The present invention is based on the seminal discovery of an M. tuberculosis gene, ldtMt2, encoding for an L,D-transpeptidase and identification of its role as a catalyst for the formation of non-classical 3→3 cross-linkages in the peptidoglycan layer. Inactivation of ldtMt2 and loss of the MT2594 protein results in altered colony morphology, attenuation in growth, loss of virulence, and increased susceptibility to β-lactams and β-lactamase inhibitors in vitro and during t

Problems solved by technology

Tuberculosis (TB) continues to be a major public health threat around the world The estimate that more lives will be lost in 2009 due to TB than in any year in history is alarming.
An increasing number of cases reporting infection with multi-(MDR) and extensively drug-resistant (XDR) strains of M. tuberculosis has diminished our capability to respond effectively against this threat.
It is speculated that poor patient compliance to treatment regimen, as the current therapy requires a combination of drugs to be taken daily for 6 months or more, is a major reason for emergence of drug resistance in TB.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Methods for treatment of bacterial infections
  • Methods for treatment of bacterial infections
  • Methods for treatment of bacterial infections

Examples

Experimental program
Comparison scheme
Effect test

example 1

Production and Purification of Recombinant LdtMt2

[0035]A portion of the ltdMt2 gene was amplified with primers 5′-TTTTCATGAT-CGCCGATCTGCTGGTGC-3′ and 5′-TTGGATCCCGCCTTGGCGTTACCGGC-3′, digested with BspHI plus BamHI (underlined) and cloned into pET2818. The resulting plasmid, encodes a fusion protein consisting of a methionine specified by the ATG initiation codon of pET2818, residues 55 to 408 of LdtMt2, and a C-terminal polyhistidine tag with the sequence GSH6. E. coli BL21 (DE3) harboring pREP4GroESL and pET2818ΩldtMt2 was grown at 37° C. to an OD600 of 0.5 in 3 L of brain heart infusion broth containing 150 μg / ml ampicillin. Expression was induced by adding Isopropyl-D-thiogalactopyranoside to a final concentration of 0.5 mM and incubating the cultures for 17 hours at 16° C. LdtMt2 was purified from a clarified lysate by affinity chromatography on Ni2+-nitrilotriacetate-agarose resin (Qiagen GmbH, Germany) followed by anion exchange chromatography (MonoQ HR5 / 5, Amersham Pharmaci...

example 2

L, D-Transpeptidase Assays

[0036]We purified disaccharide-tetrapeptide containing amidated meso-diaminopimelic acid (GlcNAc-MurNAc-L-Ala1-D-iGln2-mesoDapNH23-D-Ala4) from C. jeikeium strain CIP103337 and determined the concentration after acid hydrolysis. In vitro formation of muropeptide dimers was tested in 10 μL of 50 mM Tris-HCl (pH 7.5) containing 300 mM NaCl, 5 μM LdtMt2, and 280 μM disaccharide-tetrapeptide. The reaction mixture was incubated for two hours at 37° C. the resulting muropeptides were analyzed by nanoelectrospray tandem mass spectrometry using N2 as the collision gas.

example 3

Bacterial Strains and Culture Conditions

[0037]M. tuberculosis CDC1551, a clinical isolate, was used as the host strain and a transposon insertion mutant in MT2594 (ldtMt2::Tn) was generated, isolated and characterized as described in Lamichhane et al., PNAS, 2003, 100(12): 7213-7218. This mutant carries a Himar1 transposon insertion at +872 base from the putative translation start site of the gene. The complemented strain was generated by transforming the mutant with pGS202—2594. This is a single copy integrating plasmid based on pMH94 backbone, which was modified into a GATEWAY compatible destination vector (Invitrogen) by inserting attL1 and attL2 sites. A wild-type copy of MT2594 along with its promoter was cloned into this destination vector pGS202 to generate pGS202—2594. Genotypes of the strains were verified by Southern blotting. For in vitro growth, Middlebrook 7H9 liquid medium supplemented with 0.2% glycerol, 0.05% Tween-80, 10% vol / vol oleic acid-albumin-dextrose-catalase...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Cell angleaaaaaaaaaa
Strain pointaaaaaaaaaa
Login to View More

Abstract

The peptidoglycan layer is a vital component of the bacterial cell wall, which comprises 4→3 and 3→3 transpeptide cross-linkages, the formation of which are catalyzed by D,D- and L,D-transpeptidases, respectively. Methods for the treatment of bacterial infections with agents that inhibit L,D-transpeptidases, either alone or in combination with D,D-transpeptidase inhibitors, are provided herein. Also provided are methods for the treatment of chronic stage tuberculosis with agents that inhibit enzyme with L,D-transpeptidase activity.

Description

CROSS REFERENCE TO RELATED APPLICATION(S)[0001]This application claims the benefit of priority under 35 U.S.C. §119(e) of U.S. Ser. No. 61 / 407,256, filed Oct. 27, 2010, the entire content of which is incorporated herein by reference.GRANT INFORMATION[0002]This invention was made in part with government support under NIH Grant No. AI30036. The United States government has certain rights in this invention.BACKGROUND OF THE INVENTION[0003]1. Field of the Invention[0004]The invention relates generally to methods for treating bacterial infections by inhibition of L,D-transpeptidases and more specifically to methods for treating tuberculosis by inhibiting a Mycobacterium tuberculosis L,D-transpeptidase.[0005]2. Background Information[0006]Tuberculosis (TB) continues to be a major public health threat around the world The estimate that more lives will be lost in 2009 due to TB than in any year in history is alarming. An increasing number of cases reporting infection with multi-(MDR) and ex...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): A61K31/43A61K31/424A61P31/04A61K31/407
CPCA61K31/00A61K31/424A61K31/43A61K45/06C12Q1/18C12Q1/689C12Q2600/158C12Q2600/136C12Q1/37A61K2300/00A61P31/04
Inventor LAMICHHANE, GYANUBISHAI, WILLIAM R.GUPTA, RADHIKALAVOLLAY, MARIEMAINARDI, JEAN-LUCARTHUR, MICHEL
Owner INST NAT DE LA SANTE & DE LA RECHERCHE MEDICALE (INSERM)
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com