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

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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.

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Abbreviations

AMP:

Cationic antimicrobial peptide

BP:

Base peak

CAD:

Collision activated dissociation

CV:

Cone voltages

CYP450:

Cytochrome P450 enzyme

d P :

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

t R :

Retention time

UPLC:

Ultra performance liquid chromatography

UV:

Ultra violet

VS:

Volume standard

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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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Author notes

  1. Terkel Hansen

    Present address: Leibniz-Institut für Analytische Wissenschaften – ISAS – e.V., Otto-Hahn-Str. 6b, 44227, Dortmund, Germany

Authors and Affiliations

  1. Department of Pharmacy, Faculty of Health Sciences, University of Tromsø, 9037, Tromsø, Norway

    Terkel Hansen, Trude Anderssen & Morten B. Strøm

  2. FRAM Centre (High North Research Centre for Climate and the Environment), Norwegian Institute for Air Research, 9296, Tromsø, Norway

    Morten K. Moe

  3. Department of Multidisciplinary Laboratory Medicine and Medical Biochemistry, Akershus University Hospital, 1478, Lørenskog, Norway

    Morten K. Moe

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  1. Terkel Hansen
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Correspondence to Terkel Hansen.

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Hansen, T., Moe, M.K., Anderssen, T. et al. Metabolism of small antimicrobial β2,2-amino acid derivatives by murine liver microsomes. Eur J Drug Metab Pharmacokinet 37, 191–201 (2012). https://doi.org/10.1007/s13318-012-0086-9

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  • DOI: https://doi.org/10.1007/s13318-012-0086-9

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