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Solid-Phase Synthesis of Heterobivalent Ligands Targeted to Melanocortin and Cholecystokinin Receptors

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Abstract

Heteromultivalency provides a route to increase binding avidity and to high specificity when compared to monovalent ligands. The enhanced specificity can potentially serve as a unique platform to develop diagnostics and therapeutics. To develop new imaging agents based upon multivalency, we employed heterobivalent constructs of optimized ligands. In this report, we describe synthetic methods we have developed for the preparation of heterobivalent constructs consisting of ligands targeted simultaneously to the melanocortin receptor, hMC4R, and the cholecystokinin receptors, CCK-2R. Modeling data suggest that a linker distance span of 20–50 Å is needed to crosslink these two G-protein coupled receptors (GPCRs). The two ligands were tethered with linkers of varying rigidity and length, and flexible polyethylene glycol based PEGO chain or semi-rigid [poly(Pro-Gly)] linkers were employed for this purpose. The described synthetic strategy provides a modular way to assemble ligands and linkers on solid-phase supports. Examples of heterobivalent ligands are provided to illustrate the increased binding avidity to cells that express the complementary receptors.

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Abbreviations

GPCR:

G-protein coupled receptor

hMC4R:

Human melanocortin-4 receptor

CCK-2R:

Cholecystokinin-2 receptor

α-MSH:

α-Melanocortin stimulating hormone

MSH-7:

Ac-Ser-Nle-Glu-His-DPhe-Arg-Trp-

CCK-6:

-Nle-Gly-Trp-Nle-Asp-Phe-NH2

PEGO:

19-Amino-5-oxo-3,10,13,16-tetraoxa-6-azanonadecan-1-oic acid

Fmoc:

9-Fluorenylmethyloxycarbonyl

Boc:

Tert-butyloxycarbonyl

HOBt:

1-Hydroxybenzotriazole

HOCt:

6-Chloro-1-hydroxybenzotriazole

HBTU:

o-[1H-Benzotriazol-1-yl)(dimethylamino)methylene]uranium hexafluorophosphate N-oxide

DIC:

Diisopropylcarbodiimide

DIEA:

Diisopropylethylamine

Pbf:

2,2,4,6,7-Pentamethyldihydrobenzofuran-5-yl sulfonyl

Trt:

Triphenylmethyl (trityl)

TFA:

Trifluoroacetic acid

MALDI-TOF:

Matrix assisted laser desorption ionization-time of flight

ESI-MS:

Electrospray ionization-mass spectrometry

FT-ICR:

Fourier transform-ion cyclotron resonance

Hek-293 cells:

Human embryonic kidney cells

TRF:

Time-resolved fluorescence

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Acknowledgements

We thank Ms. Lucinda Begay and Mrs. Renata Patek for HPLC and technical assistance. This work was supported by grants R01 CA 123547 and RO1 CA097360 from the National Cancer Institute and Grant ABRC06-006 from the Arizona Biomedical Research Commission.

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Author notes

  1. Robert J. Gillies

    Present address: H. Lee Moffitt Cancer Center, 12902 Magnolia Drive, Tampa, FL, 33612, USA

Authors and Affiliations

  1. Department of Chemistry, The University of Arizona, 1306 E. University Blvd, Tucson, AZ, 85721, USA

    Jatinder S. Josan, Rajesh Sankaranarayanan & Victor J. Hruby

  2. BIO5 Institute, The University of Arizona, 1657 E. Helen Street, Tucson, AZ, 85719, USA

    Josef Vagner, Robert J. Gillies & Victor J. Hruby

  3. Department of Biochemistry and Molecular Biophysics, The University of Arizona, 1306 E. University Blvd, P.O. Box 210041, Tucson, AZ, 85721, USA

    Heather L. Handl, Robert J. Gillies & Victor J. Hruby

  4. Department of Radiology, The University of Arizona, 1306 E. University Blvd, P.O. Box 210041, Tucson, AZ, 85721, USA

    Robert J. Gillies

Authors
  1. Jatinder S. Josan
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  2. Josef Vagner
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  3. Heather L. Handl
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  4. Rajesh Sankaranarayanan
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  5. Robert J. Gillies
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  6. Victor J. Hruby
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Corresponding author

Correspondence to Victor J. Hruby.

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Josan, J.S., Vagner, J., Handl, H.L. et al. Solid-Phase Synthesis of Heterobivalent Ligands Targeted to Melanocortin and Cholecystokinin Receptors. Int J Pept Res Ther 14, 293–300 (2008). https://doi.org/10.1007/s10989-008-9150-3

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  • DOI: https://doi.org/10.1007/s10989-008-9150-3

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