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Design and synthesis of a reagent for solid-phase incorporation of the phosphothreonine mimetic (2S,3R)-2-amino-3-methyl-4-phosphonobutyric acid (Pmab) into peptides in a bio-reversible phosphonyl-bis-pivaloyloxymethyl (POM) prodrug form

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Abstract

Reported herein are the synthesis and solid-phase peptide incorporation of N-Fmoc-(2S,3R)-2-amino-3-methyl-4-phosphonobutyric acid bis-pivaloyloxymethyl phosphoryl ester [Fmoc-Pmab(POM)2-OH, 2] as a phosphatase-stable phosphothreonine (pThr) mimetic bearing orthogonal protection suitable for the synthesis of Pmab-containing peptides having bio-reversible protection of the phosphonic acid moiety. This represents the first report of a bio-reversibly protected pThr mimetic in a form suitable for facile solid-phase peptide synthesis.

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Fig. 1
Scheme 1
Scheme 2
Fig. 2

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Abbreviations

Ac:

Acetyl

[α]D :

Specific rotation

BnBr:

Benzyl bromide

But :

tert-Butyl

Cbz:

Benzyloxycarbonyl

CH2Cl2 :

Dimethyl chloride

CH3CN:

Acetonitrile

DIPEA:

Diisopropylethylamine

DMF:

Dimethylformamide

ESI:

Electrospray ionization

EtOAc:

Ethyl acetate

F2Pmp:

Difluorophosphonomethylphenylalanine

Fmoc:

9-Fluorenylmethoxycarbonyl

Fmoc-OSu:

9-Fluorenylmethyl succinimidyl carbonate

HBTU:

1-O-Benzotriazole-N,N,N′,N′-tetramethyl-uromium- hexafluoro-phosphate

HCl:

Hydrochloric acid

His:

Histidine

HOBt:

1-Hydroxybenzotriazole

HPLC:

High-performance liquid chromatography

HRMS:

High resolution mass spectra

Leu:

Leucine

LiOH:

Lithium hydroxide

MeOH:

Methanol

MgSO4 :

Magnesium sulfate

Mtt:

Methyltrityl

m/z :

Mass-to-charge ratio

NaHCO3 :

Sodium bicarbonate

NMP:

N-Methyl-2-pyrrolidone

NMR:

Nuclear magnetic resonance

PBD:

Polo box domain

Pd·C:

Palladium on carbon

PLE:

Porcine liver esterase

Plk1:

Polo-like kinase 1

Pmab:

(2S,3R)-2-Amino-3-methyl-4-phosphonobutyric acid

POM:

Pivaloyloxymethyl

POMI:

Pivaloyloxymethyl iodide

PPIs:

Protein-protein interactions

pSer:

Phosphoserine

pThr:

Phosphothreonine

pTyr:

Phosphotyrosine

Ser:

Serine

SPPS:

Solid-phase peptide synthesis

t ½ :

Half life

TFA:

Trifluoroacetic acid

THF:

Tetrahydrofuran

TLC:

Thin layer chromatography

TMS:

Trimethylsilane

Trt:

Trityl

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Acknowledgments

This research was supported by the Intramural Research Program of the NIH, Center for Cancer Research, NCI-Frederick and the National Cancer Institute, National Institutes of Health.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Chemical Biology Laboratory, Molecular Discovery Program, Center for Cancer Research, National Cancer Institute-Frederick, National Cancer Institute, National Institutes of Health, Frederick, MD, 21702, USA

    Wen-Jian Qian & Terrence R. Burke Jr.

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  1. Wen-Jian Qian
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  2. Terrence R. Burke Jr.
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Correspondence to Terrence R. Burke Jr..

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Qian, WJ., Burke, T.R. Design and synthesis of a reagent for solid-phase incorporation of the phosphothreonine mimetic (2S,3R)-2-amino-3-methyl-4-phosphonobutyric acid (Pmab) into peptides in a bio-reversible phosphonyl-bis-pivaloyloxymethyl (POM) prodrug form. Amino Acids 45, 1143–1148 (2013). https://doi.org/10.1007/s00726-013-1567-0

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