Trends in Peptide and Protein Sciences

Synthesis, Radiolabeling and Stability Studies of Peptide HYNIC-LIKKP-Pyr-F with 99mTc as an Apoptosis Imaging Agent Radiolabeling of an apoptosis imaging agent

Trends in Peptide and Protein Sciences, Vol. 7 (2022), , Page 1-5 (e8)
https://doi.org/10.22037/tpps.v7i.39024

Abstract

A non-invasive method for detecting phosphatidylserine (PS) exposure on the outer surface of plasma membranes, such as nuclear imaging, could aid in the diagnosis and treatment of diseases associated with apoptosis. Annexin V has been the most researched imaging agent for apoptosis to date. Due to Annexin V's limitations, additional agents, such as small peptides and molecules, have been introduced, including LIKKPF developed by Burtea et al. In this study, HYNIC-LIKKP-pyr-F, a derivative of LIKKPF was prepared using the 9-fluoroenylmethoxycarbonyl (fmoc) method, radiolabeled with Technetium-99m (99mTc) with the use of Stannous chloride (SnCl2) as a reducing agent and ethylenediamine diacetate (EDDA) and tricine as co-ligands. Radiochemical purity, labeling efficiency, and stability of radiopeptide in normal saline and human plasma were determined using thin layer chromatography (TLC). The partition coefficient of radiolabeled peptide was measured in a combination of PBS (pH 7.4) and n-octanol. Specific activity was also measured. LC-MS was used to examine the synthesized peptide. Peptide was stable in human serum for at least 4 hr. Peptide was radiolabeled with 99mTc with radiochemical purity and labeling efficiency over 95% and 90%, respectively. Radiopeptide was stable in saline and human serum for at least 4 hours. The radiolabeled peptide has a great deal of potential as an apoptosis imaging agent for in vitro and in vivo experiments.


HIGHLIGHTS


  • A noninvasive method for apoptosis imaging is the usage of radiolabeled affinity ligands.

  • A new derivative of LIKKPF with affinity for phosphatidyl serine was synthesized.

  • The LIKKPF peptide was radiolabeled with 99m

Keywords:
  • Apoptosis
  • 18FDG (2-deoxy-2-flouro-D-glucose)
  • 6-Hydrazinonicotinamide (HYNIC)
  • LIKKP-Pyr-F
  • Phosphatidylserine
  • Radiopeptide
  • Technetium-99m (99mTc)

How to Cite

1.
Javani Jouni E, Kordi M, Khoshbakht S, Amidi S, Shahhosseini S. Synthesis, Radiolabeling and Stability Studies of Peptide HYNIC-LIKKP-Pyr-F with 99mTc as an Apoptosis Imaging Agent : Radiolabeling of an apoptosis imaging agent. Trends Pept. Protein Sci. [Internet]. 2022Sep.13 [cited 2023Mar.23];7:1-5 (e8). Available from: https://journals.sbmu.ac.ir/protein/article/view/39024

References

Blankenberg, F.G., Katsikis, P.D., Tait, J.F., Davis, R.E., Naumovski, L., Ohtsuki, K., Kopiwoda, S., Abrams, M.J., Darkes, M., Robbins, R.C. and H.T. Maecker, (1998). ″In vivo detection and imaging of phosphatidylserine expression during programmed cell death.″ Proceedings of the National Academy of Sciences, 95(11): 6349-6354. DOI: https://doi.org/10.1073/pnas.95.11.6349.

Blankenberg, F.G., Tait, J., Ohtsuki, K. and H.W. Strauss, (2000). ″Apoptosis: the importance of nuclear medicine.″ Nuclear Medicine Communications, 21(3): 241-250. DOI: https://doi.org/10.1097/00006231-200003000-00008.

Blankenberg, F.G. (2008). ″In vivo detection of apoptosis.″ Journal of Nuclear Medicine, 49(Suppl 2): 81S-95S. DOI: https://doi.org/10.2967/jnumed.107.045898.

Burtea, C., Laurent, S., Lancelot, E., Ballet, S., Murariu, O., Rousseaux, O., Port, M., Vander Elst, L., Corot, C. and R.N. Muller, (2009). ″Peptidic targeting of phosphatidylserine for the MRI detection of apoptosis in atherosclerotic plaques.″ Molecular pharmaceutics, 6(6): 1903-1919. DOI: https://doi.org/10.1021/mp900106m.

Hanshaw, R.G. and B.D. Smith, (2005). ″New reagents for phosphatidylserine recognition and detection of apoptosis.″ Bioorganic & medicinal chemistry, 13(17): 5035-5042. DOI: https://doi.org/10.1016/j.bmc.2005.04.071.

Khoshbakht, S., Beiki, D., Geramifar, P., Kobarfard, F., Sabzevari, O., Amini, M. and S. Shahhosseini, (2016a). ″18FDG-labeled LIKKPF: a PET tracer for apoptosis imaging.″ Journal of Radioanalytical and Nuclear Chemistry, 310(1): 413-421. DOI: https://doi.org/10.1007/s10967-016-4793-6.

Khoshbakht, S., Kobarfard, F., Beiki, D., Sabzevari, O., Amini, M., Mehrnejad, F., Tabib, K. and S. Shahhosseini, (2016b). ″HYNIC a bifunctional prosthetic group for the labelling of peptides with 99mTc and 18FDG.″ Journal of Radioanalytical and Nuclear Chemistry, 307(2): 1125-1134. DOI: https://doi.org/10.1007/s10967-015-4259-2.

Khoshbakht, S., Beiki, D., Geramifar, P., Kobarfard, F., Sabzevari, O., Amini, M., Mehrnejad, F. and S. Shahhosseini, (2016c). ″Synthesis, radiolabeling, and biological evaluation of peptide LIKKPF functionalized with HYNIC as apoptosis imaging agent.″ Iranian Journal of Pharmaceutical Research: IJPR, 15(2): 415-424. PMID: 27642312.

Khoshbakht, S., Beiki, D., Geramifar, P., Kobarfard, F., Sabzevari, O., Amini, M., Bolourchian, N., Shamshirian, D. and S. Shahhosseini, (2019). ″Design, synthesis, radiolabeling, and biologic evaluation of three 18F-FDG-radiolabeled targeting peptides for the imaging of apoptosis.″ Cancer Biotherapy & Radiopharmaceuticals, 34(5):271-279. DOI: https://doi.org/10.1089/cbr.2018.2709.

Li, X., Link, J.M., Stekhova, S., Yagle, K.J., Smith, C., Krohn, K.A. and J.F. Tait, (2008). ″Site-specific labeling of annexin V with F-18 for apoptosis imaging.″ Bioconjugate chemistry, 19(8): 1684-1688. DOI: https://doi.org/10.1021/bc800164d.

Makin, G. and C. Dive, (2001). ″Apoptosis and cancer chemotherapy.″ Trends in Cell Biology, 11: S22-S26. DOI: https://doi.org/10.1007/s004419900160.

Mosayebnia, M., Hajiramezanali, M. and S. Shahhosseini, (2020). ″Radiolabeled peptides for molecular imaging of apoptosis.″ Current Medicinal Chemistry, 27(41): 7064-7089. DOI: https://doi.org/10.2174/0929867327666200612152655.

Post, A.M., Katsikis, P.D., Tait, J.F., Geaghan, S.M., Strauss, H.W. and F.G. Blankenberg, (2002). ″Imaging cell death with radiolabeled annexin V in an experimental model of rheumatoid arthritis.″ Journal of Nuclear Medicine, 43(10): 1359-1365. PubMed: 12368374.

Schibli, R. and A.P. Schubiger, (2002). ″Current use and future potential of organometallic radiopharmaceuticals.″ European Journal of Nuclear Medicine and Molecular Imaging, 29(11): 1529-1542. DOI: https://doi.org/10.1007/s00259-002-0900-8.

Schutters, K. and C. Reutelingsperger, (2010). ″Phosphatidylserine targeting for diagnosis and treatment of human diseases.″ Apoptosis, 15(9): 1072-1082. DOI: https://doi.org/10.1007/s10495-010-0503-y .

Stace, C.L. and N.T. Ktistakis, 2006. Phosphatidic acid-and phosphatidylserine-binding proteins. Biochimica et Biophysica Acta (BBA)-Molecular and Cell Biology of Lipids, 1761(8): 913-926. DOI: https://doi.org/10.1016/j.bbalip.2006.03.006 .

Thapa, N., Kim, S., So, I.S., Lee, B.H., Kwon, I.C., Choi, K. and I.S. Kim, (2008). ″Discovery of a phosphatidylserine‐recognizing peptide and its utility in molecular imaging of tumour apoptosis.″ Journal of Cellular and Molecular Medicine, 12(5a): 1649-1660. DOI: https://doi.org/10.1111/j.1582-4934.2008.00305.x.

Zhao, M., Li, Z. and S. Bugenhagen, (2008). ″99mTc-labeled duramycin as a novel phosphatidylethanolamine-binding molecular probe.″ Journal of Nuclear Medicine, 49(8): 1345-1352. DOI: https://doi.org/10.2967/jnumed.107.048603.

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