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Synthesis and Biological Evaluation of Cyclic [99mTc]-HYNIC-CGPRPPC as a Fibrin-Binding Peptide for Molecular Imaging of Thrombosis and Its Comparison with [99mTc]-HYNIC-GPRPP

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Abstract

Purpose

Many patients worldwide suffer from cardiovascular diseases for which an underlying factor is thrombosis. Devising a molecular imaging technique for early detection of thrombosis in a clinical setting is highly recommended. Because fibrin is a major constituent of clots and is present in all types of thrombi but absent in circulation, it is a highly specific and sensitive target for molecular imaging of thrombi. It is assumed that cyclization of peptides will improve the receptor binding affinity and stability of the peptide. In the present study, we have developed linear and cyclic fibrin-binding peptides for thrombus imaging and compared their biological properties.

Procedures

Linear HYNIC-GPRPP and cyclic HYNIC-CGPRPPC peptides were synthesized using a standard Fmoc strategy and radiolabeled with Tc-99m. The stability of the radiolabeled peptides in human plasma and their affinity for fibrin and blood clots were determined. Blood clearance and biodistribution were evaluated in rats and mice, respectively. The peptide with the highest affinity was injected to a live rabbit femoral thrombosis model, and scintigraphic images were obtained.

Results

In vitro studies show that peptides are stable in human plasma and have a high affinity for human fibrin. They also demonstrated fast blood clearance in rats and high thrombus uptake in the Balb/c mice femoral thrombosis model. Femoral thrombosis was visualized 30 min postinjection of cyclic peptide in a live rabbit model using single photon emission computed tomography (SPECT)/X-ray computed tomography.

Conclusions

The results indicate that the cyclic peptide is a promising agent for molecular imaging of fibrin using SPECT.

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Abbreviations

GPRPP:

Glycine proline arginine proline proline

CGPRPPC:

Cysteine glycine proline arginine proline proline cysteine

HYNIC:

6-Hydrazinonicotinamide

TFA:

Trifluoroacetic acid

Tricine:

N-[Tris(hydroxymethyl)methyl]glycine

DIPEA:

Diisopropylethylamine

DMF:

Dimethylformamide

TIS:

Triisobutylsilane

MEK:

Methyl ethyl ketone

EDDA:

Ethylenediamine diacetate

Fmoc:

9-Fluoroenylmethoxycarbonyl

DCM:

Dichloromethane

MeOH:

Methanol

HBS:

HEPES-buffered saline

DVT:

Deep venous thrombosis

PE:

Pulmonary embolism

TBTU:

O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate

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Acknowledgments

This research was supported by Shahid Beheshti University of Medical Sciences, Tehran, Iran; grant no. 8002.

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Authors and Affiliations

  1. Department of Chemistry, North Tehran Branch of Islamic Azad University, Tehran, Iran

    Sedigheh Rezaeianpour & Abolghasem Moghimi

  2. Alborz University of Medical Sciences, Karaj, Iran

    Atefeh Hajiagha Bozorgi

  3. Department of Pharmaceutics, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Ameneh Almasi

  4. Peptide Chemistry Research Center, K.N. Toosi University of Technology, Tehran, Iran

    Saeed Balalaie & Sorour Ramezanpour

  5. Department of Pharmacology and Toxicology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Sanaz Nasoohi

  6. Radiation Application Research School, Nuclear Science and Technology Research Institute, Tehran, Iran

    Seyed Mohammad Mazidi

  7. Research Center for Nuclear Medicine, Tehran University of Medical Sciences, Tehran, Iran

    Parham Geramifar

  8. Cardiovascular Interventional Research Center, Department of Nuclear Medicine, Rajaei Cardiovascular, Medical and Research Center, Iran University of Medical Sciences, Tehran, Iran

    Ahmad Bitarafan-Rajabi

  9. Department of Pharmaceutical Chemistry and Radiopharmacy, School of Pharmacy and PET/CT Unit, Ferdous Nuclear Medicine Center, Dr. Masih Daneshvari Hospital, Shahid Beheshti University of Medical Sciences, Vali-e Asr Ave., Niayesh Junction, P.O. Box 14155-6153, Tehran, Iran

    Soraya Shahhosseini

Authors
  1. Sedigheh Rezaeianpour
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  2. Atefeh Hajiagha Bozorgi
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  10. Ahmad Bitarafan-Rajabi
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  11. Soraya Shahhosseini
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Corresponding author

Correspondence to Soraya Shahhosseini.

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The authors declare that they have no conflict of interest.

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Rezaeianpour, S., Bozorgi, A.H., Moghimi, A. et al. Synthesis and Biological Evaluation of Cyclic [99mTc]-HYNIC-CGPRPPC as a Fibrin-Binding Peptide for Molecular Imaging of Thrombosis and Its Comparison with [99mTc]-HYNIC-GPRPP. Mol Imaging Biol 19, 256–264 (2017). https://doi.org/10.1007/s11307-016-1004-3

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  • DOI: https://doi.org/10.1007/s11307-016-1004-3

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