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Metabolism of small antimicrobial β2,2-amino acid derivatives by murine liver microsomes

  • Terkel HansenEmail author
  • Morten K. Moe
  • Trude Anderssen
  • Morten B. Strøm
Original Paper

Abstract

We have investigated the in vitro metabolism of three small antimicrobial β2,2-amino acid derivatives (M w < 500) that are highly potent against methicillin resistant Staphylococcus aureus, and are among the first compounds designed from small cationic antimicrobial peptides with potential for oral administration. The β2,2-amino acid derivatives are virtually completely resistant against degradation by proteases, and to further explore their drug potential, we have investigated the hepatic Phase I metabolism of this class of antimicrobial compounds. The β2,2-amino acid derivatives were incubated with murine liver microsomes and the metabolites analyzed semi-quantitatively by HPLC–MS and qualitatively by ultra performance liquid chromatography coupled to a tandem mass spectrometer which enabled identification of the metabolites by careful interpretation of the collision activated dissociation spectra. The study shows that sterically hindered β2,2-amino acid derivatives that otherwise are stable against proteolytic degradation underwent Phase I metabolism and were oxidized to a number of different metabolites depending on the structure of the β2,2-amino acid side-chains.

Keywords

In vitro metabolism Antimicrobial β2,2-amino acid derivatives AMPs Scaffold Phase I metabolism 

Abbreviations

AMP

Cationic antimicrobial peptide

BP

Base peak

CAD

Collision activated dissociation

CV

Cone voltages

CYP450

Cytochrome P450 enzyme

dP

Particle diameter

ESI

Electrospray ionization

HPLC

High performance liquid chromatography

IS

Internal standard

LoD

Limit of detection

LoQ

Limit of quantification

MIC

Minimal inhibitory concentration

MNP

Methylene naphthalene

MRSA

Methicillin resistant Staphylococcus aureus

MRSE

Methicillin resistant Staphylococcus epidermidis

MS

Mass spectrometry

NADP

Nicotinamide adenine dinucleotide phosphate

NL

Neutral loss

Q-ToF

Quadrupole time of flight

RBC

Human erythrocytes

RP

Reverse phase

SEM

Standard error of the mean

SIR

Single-ion recording

ToF

Time of flight

tR

Retention time

UPLC

Ultra performance liquid chromatography

UV

Ultra violet

VS

Volume standard

Notes

Acknowledgments

We thank Dr. Jack Bruun for technical assistance with running the Q-ToF experiments as well as Dr. Espen Hansen for help with obtaining the HRMS spectra.

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Copyright information

© Springer-Verlag France 2012

Authors and Affiliations

  • Terkel Hansen
    • 1
    • 4
    Email author
  • Morten K. Moe
    • 2
    • 3
  • Trude Anderssen
    • 1
  • Morten B. Strøm
    • 1
  1. 1.Department of Pharmacy, Faculty of Health SciencesUniversity of TromsøTromsøNorway
  2. 2.FRAM Centre (High North Research Centre for Climate and the Environment)Norwegian Institute for Air ResearchTromsøNorway
  3. 3.Department of Multidisciplinary Laboratory Medicine and Medical BiochemistryAkershus University HospitalLørenskogNorway
  4. 4.Leibniz-Institut für Analytische Wissenschaften – ISAS – e.V.DortmundGermany

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