Cadherins are cell adhesion and cell signaling molecules that provide the molecular link between each adjacent cells and have critical importance for the initiation and continuation of adhesion mechanism. N-cadherin expression provides a correlation between upregulation of N-cadherin and inflammation of the lesions. In this paper, we concentrated on the radiolabeling and evaluation of [68Ga]Ga-AntiCAD1 agent as a potential candidate for in vivo PET/CT imaging of adhesions. The synthetic N-Ac-CHAVC-NH2 cyclic peptide sequence designed as (ADH-1)c containing the selective binding His-Ala-Val (HAV) motif based on the chimeric antigen receptor sequence acts as the N-cadherin antagonist. In our previous study, AntiCAD1 conjugate has been studied in detail which is in the process of publication. In this study, the conjugate was radiolabelled with the [68Ga]Ga radionuclide eluted from the 68Ge/68Ga generator (IDB Holland). Radiochemical purity of [ 68Ga]Ga-AntiCAD1 agent was analysed with TLC methods. The ‘shake-flask’ method was applied to determine lipophilicity of the agent by calculating the P distribution coefficient (logP=-2.69±0.54). The biodistribution of the agent was investigated using PET/CT on Wistar Albino rats. Significant uptake was found in liver, kidneys, spleen, salivary gland and targeted region with SUVmax-mean of 1.36, 1.96, 1.38, 1.16 and 2.14 respectively. The Pearson Factorial method is used to test the relationship between the targeted region and other body tissues, to measure the degree of this relationship (R=0.73). Radiolabelled agent was demonstrated to react specifically with N-cadherin in targeting of rat tissues.
Kadherinler, bitişik hücreler arasındaki moleküler bağlantıyı sağlayan, yapışma mekanizmasının başlatılması ve devamı için kritik öneme sahip olan hücre adezyon ve hücre sinyal molekülleridir. N-kadherin ekspresyonu, N-kadherinin artışı ve lezyonların inflamasyonu arasında bir korelasyon sağlar. Bu makalede, adezyonların in vivo PET/CT görüntülenmesi için potansiyel bir aday olarak [68Ga]Ga-AntiCAD1 ajanının radyoişaretlenmesi ve değerlendirmesine odaklanılmıştır. Kimerik antijen reseptörü dizisine dayanan seçici bağlayıcı His-Ala-Val (HAV) motifini içeren (ADH-1)c olarak tasarlanan sentetik N-Ac-CHAVC-NH2 siklik peptid dizisi, N-kadherin antagonisti olarak işlev görür. Yayın aşamasındaki önceki çalışmamızda AntiCAD1 konjugatı ayrıntılı olarak incelenmiştir. Bu çalışmamızda, konjugat, 68Ge/68Ga jeneratöründen (IDB Holland) elde edilen [68Ga]Ga radyonüklidi ile radyoaktif olarak işaretlendi. [68Ga]Ga-AntiCAD1 ajanının radyokimyasal saflığı TLC metodları ile analiz edildi. Ajanın lipofilisitesini belirlemek için P dağılım katsayısı hesaplanarak ‘çalkalama şişesi’ yöntemi uygulandı (logP=- 2.69±0.54). Ajanın biyodağılımı, Wistar Albino sıçanları üzerinde PET/CT kullanılarak araştırıldı. Karaciğer, böbrekler, dalak, tükürük bezi ve hedef bölgede sırasıyla ortalama 1.36, 1.96, 1.38, 1.16 ve 2.14 SUVmax değerleri ile kayda değer oranda tutulum tespit edildi. Hedeflenen bölge ile diğer vücut dokuları arasında olan ilişkini test etmek ve bu ilişkinin derecesini ölçmek için istatistiksel Pearson Faktörü hesaplama yöntemi kullanılmıştır (R=0.73). Sıçan dokularının hedeflenmesinde radyoişaretli ajanın N-kadherin ile spesifik olarak etkileşime girdiği gösterildi.
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Klein W, Kördel W, Weiss M, Poremski HJ: Updating of the OECD test guideline 107 “partition coefficient n-octanol/water”: OECD laboratory intercomparison test on the HPLC method. Chemosphere, 17 (2): 361-386, 1988. DOI: 10.1016/0045-6535(88)90227-5
Umbricht CA, Benešová M, Schmid RM, Turler A, Schibli R, van der Meulen NP, Müller C: 44Sc-PSMA-617 for radiotheragnostics in tandem with 177Lu-PSMA-617-preclinical investigations in comparison with 68GaPSMA-11 and 68Ga-PSMA-617. EJNMMI Res, 7 (9): 1-10, 2017. DOI: 10.1186/ s13550-017-0257-4
Ertay T, Unak P, Sencan Eren M, Biber Muftuler FZ, Ozdoğan O, Ulker O, Yesilagac R, Durak H: A new 99mTc labeled peptide: 99mTc β-casomorphin 6, biodistribution and imaging studies on rats. Kafkas Univ Vet Fak Derg, 23 (1): 15-22, 2017. DOI: 10.9775/kvfd.2016.15691
Wester HJ: Pharmaceutical radiochemistry (I) - (Munich Molecular Imaging Handbook Series. Vol. 1. 198-200, Scintomics, Munich, 2010.
Leo A, Hansch C, Elkins D: Partition coefficients and their uses. Chem Rev, 71 (6): 525-616, 1971. DOI: 10.1021/cr60274a001
Rubas W, Cromwell MEM: The effect of chemical modifications on octanol/water partition (log D) and permeabilities across Caco-2 monolayers. Adv Drug Deliv Rev, 23 (1-3): 157-162, 1997. DOI: 10.1016/ S0169-409X(96)00433-4
Berthod A, Carda-Broch S: Determination of liquid-liquid partition coefficients by separation methods. J Chromatogr A, 1037 (1-2): 3-14, 2004. DOI: 10.1016/j.chroma.2004.01.001
Wilson AA, Jin L, Garcia A, DaSilva JN, Houle S: An admonition when measuring the lipophilicity of radiotracers using counting techniques. Appl Radiat Isot, 54 (2): 203-208, 2001. DOI: 10.1016/S0969- 8043(00)00269-4
Waterhouse RN: Determination of lipophilicity and its use as a predictor of blood-brain barrier penetration of molecular imaging agents. Mol Imaging Biol, 5 (6): 376-389, 2003. DOI: 10.1016/j.mibio. 2003.09.014
Pike VW: PET radiotracers: Crossing the blood-brain barrier and surviving metabolism. Trends Pharmacol Sci, 30 (8): 431-440, 2009. DOI: 10.1016/j.tips.2009.05.005
Baur B, Solbach C, Andreolli E, Winter G, Machulla HJ, Reske SN: Synthesis, radiolabelling and in vitrocharacterization of the Gallium-68-, Yttrium-90-and Lutetium-177-Labelled PSMA Ligand, CHX-A’’-DTPADUPA-Pep. Pharmaceuticals, 7 (5): 517-529, 2014. DOI: 10.3390/ph7050517
Sarikaya M, Enginar H: Radioabeling of L-tyrosine with 131I and investigation of radiopharmaceutical potantial. AKU J Sci Eng,12, 1-9, 2012.
Kızılbey K, Mansuroglu B, Derman S, Battal YB, Mustafaeva Z: Conjugation of BSA protein and VP/AA Copolymers. IJNES, 3 (2): 36-40, 2009.
Deliloglu Gurhan SI, Akdeste MM, Akdeste Mustafaeva Z, Aynagoz G, Unver G, Unal N, Celik N: Peparation of synthetic peptide FMD vaccine with newly developed antigen- polymere conjugates be used as immonogen and vaccine in veterinary medicine. Report of the Europ Comm Cont FMD, Sess Res Gr Stand Tech Com, Çeşme, Izmir, Turkey, FAO/UN Rome, 17-20 September 2002.www.fao.org/ag/againfo/ commissions/docs/research_group/izmir/App41.pdf
Mustafaeva Z: Synthesis and characterization of antigenic peptide of sheep pox disease. Kimya Problemlеri, 3, 255-263, 2017.
Dubey N, Varshney R, Shukla J, Ganeshpurkar A, Hazari PP, Bandopadhaya GP, Mishra AK, Trivedi P: Synthesis and evaluation of biodegradable PCL/PEG nanoparticles for neuroendocrine tumor targeted delivery of somatostatin analog. Drug Deliv, 19 (3): 132-142, 2012. DOI: 10.3109/10717544.2012.657718
Palasek SA, Cox ZJ, Collins JM: Limiting racemization and aspartimide formation in microwave‐enhanced Fmoc solid phase peptide synthesis. J Pept Sci, 13 (3): 143-148, 2007. DOI: 10.1002/psc.804
Ozdemir ZO, Topuzogulları M, Karabulut E, Akdeste ZM: Characterization and purification of viral peptides synthesized with microwave assisted solid phase method. IJNES, 3 (2): 41-44, 2009.
Ozdemir ZO, Karahan M, Karabulut E, Mustafaeva Z: Characterization of foot-and-mouth disease virus’s viral peptides with LC-ESI-MS. J Chem Soc Pakistan, 32 (4): 531-536, 2010.
Velikyan I: The diversity of 68Ga-based imaging agents. In, Baum RP, Rösch F (Eds): Theranostics, Gallium-68, and other radionuclides: A pathway to personalized diagnosis and treatment. 101-131, Springer, Berlin, 2013.
Varshney R, Hazari PP, Fernandez P, Schulz J, Allard M, Mishra AK: 68Ga-labeled bombesin analogs for receptor-mediated imaging. In, Baum RP, Rösch F (Eds): Theranostics, Gallium-68, and other radionuclides: A pathway to personalized diagnosis and treatment. 211-256, Springer, Berlin, 2013.
Lundqvist H, Tolmachev V: Targeting peptides and positron emission tomography. Biopolymers, 66 (6): 381-392, 2002. DOI: 10.1002/bip.10348
Nieman MT, Prudoff RS, Johnson KR, Wheelock MJ: N-cadherin promotes motility in human breast cancer cells regardless of their E-cadherin expression. J Cell Biol, 147 (3): 631-644, 1999. DOI: 10.1083/ jcb.147.3.631
Keller C, Kroening S, Zuehlke J, Kunath F, Krueger B, GoppeltStruebe M: Distinct mesenchymal alterations in N-cadherin and E-cadherin positive primary renal epithelial cells. PLoS One, 7 (8): e43584, 2012. DOI: 10.1371/journal.pone.0043584
Mariotti A, Perotti A, Sessa C, Rüegg C: N-cadherin as a therapeutic target in cancer. Expert Opin Investig Drugs, 16 (4): 451-465, 2007. DOI: 10.1517/13543784.16.4.451
Blaschuk OW, Gour BJ, Farookhi R, Ali A, George S: Adherex Technologies Inc. Compounds and methods for modulating endothelial cell adhesion. CanadaUS20060183884A1. 2003.
Blaschuk OW, Gour BJ: McGill University. Compounds and methods for modulating cell adhesion. Canada US6169071B1. 2006.
Bobryshev YV, Cherian SM, Inder SJ, Lord RSA: Neovascular expression of VE-cadherin in human atherosclerotic arteries and its relation to intimal inflammation. Cardiovasc Res, 43 (4): 1003-1017, 1999. DOI: 10.1016/S0008-6363(99)00125-X
Fosgerau K, Hoffmann T: Peptide therapeutics: Current status and future directions. Drug Discov Today, 20 (1): 122-128, 2015. DOI: 10.1016/j. drudis.2014.10.003
Blaschuk OW, Devemy E: Cadherins as novel targets for anti-cancer therapy. Eur J Pharmacol, 625 (1-3): 195-198, 2009. DOI: 10.1016/j. ejphar.2009.05.033
Gerhardt H, Wolburg H, Redies C: N‐cadherin mediates pericytic‐ endothelial interaction during brain angiogenesis in the chicken. Dev Dyn, 218 (3): 472-479, 2000. DOI: 10.1002/1097-0177(200007)218:3<472::AIDDVDY1008>3.0.CO;2-#
Citi S: The molecular organization of tight junctions. J Cell Biol, 121 (3): 485-489, 1993. DOI: 10.1083/jcb.121.3.485