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Molecular Imaging and Contrast Agent Database (MICAD) [Internet]. Bethesda (MD): National Center for Biotechnology Information (US); 2004-2013.

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64Cu-1,4,7,10-Tetraazacyclododecane-1,4,7,10-tetraacetic acid-Arg-Arg-Natl-Cys-Tyr-Cit-Lys-d-Lys-Pro-Tyr-Arg-Cit-Cys-Arg-NH2

, PhD
National Center for Biotechnology Information, NLM, NIH, Bethesda, MD
Corresponding author.

Created: ; Last Update: February 9, 2012.

Chemical name:64Cu-1,4,7,10-Tetraazacyclododecane-1,4,7,10-tetraacetic acid-Arg-Arg-Natl-Cys-Tyr-Cit-Lys-d-Lys-Pro-Tyr-Arg-Cit-Cys-Arg-NH2
Abbreviated name:64Cu-DOTA-NFB
Agent category:Peptide
Target:Chemokine receptor 4 (CXCR4)
Target category:Receptor
Method of detection:Positron emission tomography (PET)
Source of signal:64Cu
  • Checkbox In vitro
  • Checkbox Rodents
Structure not available in PubChem.



Chemokine receptors are G-protein–coupled receptors directing cell movement toward higher concentrations of chemokines. Chemokine receptor 4 (CXCR4) and its ligand, stromal cell–derived factor-1 (SDF-1 or CXCL12), are known to play a major role in the migration of progenitor cells during embryonic development of the central nervous, cardiovascular, and hematopoietic systems (1, 2). In addition, this CXCR4-SDF-1 receptor system has a function in the development, progression, and spread of various cancers (3), and CXCR4 acts as a co-receptor for human immunodeficiency virus (HIV) on CD4+ T cells (4). It has been suggested that CXCR4/SDF-1 interaction contributes to the pathogenesis of neurodegenerative and inflammatory conditions (5). CXCR4 is expressed by many different types of cancers, and overexpression of CXCR4 in cancers indicates poor prognosis with aggressive and metastatic tumors and resistance to chemotherapy (6).

CXCR4 is considered to play an important role in HIV infections and cancers. It is critical to perform imaging studies to measure CXCR4 levels under in vivo conditions for various pathological and physiological conditions (7). 99mTc-SDF-1 has been used with single-photon emission computed tomography (SPECT) to determine changes in CXCR4 expression in the heart after a myocardial infarction. 64Cu-1,1'-[1,4-Phenylenebis(methylene)]-bis[1,4,8,11-tetraaza-cyclotetradecane] (64Cu-AMD3100), an inhibitor of CXCR4 activity, has been studied with positron emission tomography (PET) (8). An 111In-labeled CXCR4 antagonist peptide, 111In-Ac-TZ14011, has been developed for SPECT imaging of CXCR4 expression in xenograft tumors in mice (9). Tamamura et al. (10) identified TN14003, a 14-amino-acid peptide (Arg-Arg-Natl-Cys-Tyr-Cit-Lys-d-Lys-Pro-Tyr-Arg-Cit-Cys-Arg-NH2) with high anti-HIV and CXCR4 antagonistic activities in vitro. Jacobson et al. (11) radiolabeled TN14003 with 4-[18F]fluorobenzoic acid ([18F]FBA) to form 4-[18F]fluorobenzoyl-Arg-Arg-Natl-Cys-Tyr-Cit-Lys-d-Lys-Pro-Tyr-Arg-Cit-Cys-Arg-NH2 (4-[18F]F-T140) for PET imaging of tumor CXCR4 expression with a high tumor/background ratio. However, 4-[18F]F-T140 exhibited high nonspecific accumulation in red blood cells (RBCs), which masked its binding to CXCR4 on tumors. Later, Jacobson et al. (12) replaced 4-fluorobenzoyl in T140 with 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-1,4,7,10-tetraacetic acid (DOTA) to form DOTA-NFB or 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA) to form NOTA-NFB for radiolabeling with 64Cu. 64Cu-DOTA-NFB and 64Cu-NOTA-NFB showed higher tumor/background ratios than 4-[18F]F-T140 with lower bindings to RBCs.



Jacobson et al. (12) incubated TN14003 (0.84 µmol) and DOTA-N-hydroxysuccinimide ester (1.01 µmol) in dimethylformamide and diisopropylethylamine for 18 h at 4°C. DOTA-NFB was obtained with 50%–62% chemical yield after purification with high-performance liquid chromatography (HPLC). The structure of DOTA-NFB was confirmed with HPLC and mass spectroscopy analysis supporting a 1:1 DOTA to NFB conjugation. Radiolabeling was performed by mixing 370–444 MBq (10–12 mCi) 64Cu-acetate with DOTA-NFB (12 nmol) in ammonium acetate buffer at pH 5.5. The mixture was heated for 20 min at 40°C. 64Cu-DOTA-NFB was purified with HPLC. The radiochemical purity was 99%, with a specific activity of 37 MBq/nmol (1 mCi/nmol) at the end of purification. The labeling yield was 91 ± 3% (n =5), with a total synthesis time of 50–55 min.

In Vitro Studies: Testing in Cells and Tissues


Chinese hamster ovary (CHO) cells were transfected with human CXCR4 (7 × 105 binding sites/cell). A receptor-binding assay was performed with 125I-SDF-1. T140 exhibited a 50% inhibition concentration (IC50) value of 2.5 nM (11), whereas DOTA-NFB showed a IC50 value of 68 nM (12). T140 also exhibited a lower IC50 value than DOTA-NFB to inhibit Jurkat T cell migration in response to SDF-1.

Animal Studies



Jacobson et al. (12) performed ex vivo biodistribution studies of 1.85 MBq (0.05 mCi) 64Cu-DOTA-NFB in nude mice (n = 5) bearing CHO and CHO-CXCR4 tumor xenografts at 4 h after injection. Accumulation in the spleen (CXCR4-expressing organ) was 6.7 ± 0.8% injected dose/gram (ID/g). The blood accumulation was low at <0.5% ID/g. Accumulation in the liver (18% ID/g) and kidneys (60% ID/g) was higher than in the spleen. The accumulation of radioactivity was significantly higher (P > 0.05) in the CHO-CXCR4 tumors (5.0 ± 0.9% ID/g) than in the CXCR4-negative CHO tumors (~1% ID/g). Co-injection of excess DOTA-NFB (20 nmol) significantly reduced (P < 0.01) the accumulation of radioactivity in the spleen and CXCR4-positive tumors but had little effect in the blood, muscle, liver, and CHO tumors. On the other hand, the accumulation in the kidneys increased to ~80% ID/g. The tumor/blood and tumor/muscle ratios were 14.5 ± 0.8 and 19.3 ± 2.4, respectively, at 4 h after injection of only 64Cu-DOTA-NFB. Co-injection with 20 nmol DOTA-NFB significantly decreased the tumor/blood and tumor/muscle ratios to 3.8 ± 0.2 and 8.2 ± 2.7 (P < 0.01), respectively.

The whole-body distribution of 64Cu-DOTA-NFB was assessed with static PET imaging at 1, 2, 4, and 24 h after injection in nude mice (n = 5) bearing CHO and CHO-CXCR4 tumor xenografts (12). The CHO-CXCR4 tumors were clearly visualized along with the abdomen area at 1 h after injection, but the CHO tumors were not. The accumulation in the CHO-CXCR4 tumors was constant over time at ~4% ID/g. There was little radioactivity (<0.5% ID/g) in the blood and muscle. The liver and kidneys exhibited slow washout, with 13.6% ID/g and 22.4% ID/g at 24 h, respectively.

Other Non-Primate Mammals


No publication is currently available.

Non-Human Primates


No publication is currently available.

Human Studies


No publication is currently available.

NIH Support

Intramural Research Program


Juarez J., Bendall L. SDF-1 and CXCR4 in normal and malignant hematopoiesis. Histol Histopathol. 2004;19(1):299–309. [PubMed: 14702198]
Miller R.J., Banisadr G., Bhattacharyya B.J. CXCR4 signaling in the regulation of stem cell migration and development. J Neuroimmunol. 2008;198(1-2):31–8. [PMC free article: PMC4448969] [PubMed: 18508132]
Rubin J.B. Chemokine signaling in cancer: one hump or two? Semin Cancer Biol. 2009;19(2):116–22. [PMC free article: PMC2694237] [PubMed: 18992347]
Alkhatib G. The biology of CCR5 and CXCR4. Curr Opin HIV AIDS. 2009;4(2):96–103. [PMC free article: PMC2718543] [PubMed: 19339947]
Mocchetti I., Bachis A., Masliah E. Chemokine receptors and neurotrophic factors: potential therapy against aids dementia? J Neurosci Res. 2008;86(2):243–55. [PubMed: 17847079]
Gelmini S., Mangoni M., Serio M., Romagnani P., Lazzeri E. The critical role of SDF-1/CXCR4 axis in cancer and cancer stem cells metastasis. J Endocrinol Invest. 2008;31(9):809–19. [PubMed: 18997494]
Misra P., Lebeche D., Ly H., Schwarzkopf M., Diaz G., Hajjar R.J., Schecter A.D., Frangioni J.V. Quantitation of CXCR4 expression in myocardial infarction using 99mTc-labeled SDF-1alpha. J Nucl Med. 2008;49(6):963–9. [PMC free article: PMC2712574] [PubMed: 18483105]
Jacobson O., Weiss I.D., Szajek L., Farber J.M., Kiesewetter D.O. 64Cu-AMD3100--a novel imaging agent for targeting chemokine receptor CXCR4. Bioorg Med Chem. 2009;17(4):1486–93. [PMC free article: PMC2723765] [PubMed: 19188071]
Hanaoka H., Mukai T., Tamamura H., Mori T., Ishino S., Ogawa K., Iida Y., Doi R., Fujii N., Saji H. Development of a 111In-labeled peptide derivative targeting a chemokine receptor, CXCR4, for imaging tumors. Nucl Med Biol. 2006;33(4):489–94. [PubMed: 16720240]
Tamamura H., Xu Y., Hattori T., Zhang X., Arakaki R., Kanbara K., Omagari A., Otaka A., Ibuka T., Yamamoto N., Nakashima H., Fujii N. A low-molecular-weight inhibitor against the chemokine receptor CXCR4: a strong anti-HIV peptide T140. Biochem Biophys Res Commun. 1998;253(3):877–82. [PubMed: 9918823]
Jacobson O., Weiss I.D., Kiesewetter D.O., Farber J.M., Chen X. PET of tumor CXCR4 expression with 4-18F-T140. J Nucl Med. 2010;51(11):1796–804. [PMC free article: PMC3629977] [PubMed: 20956475]
Jacobson, O., I.D. Weiss, L.P. Szajek, G. Niu, Y. Ma, D.O. Kiesewetter, A. Peled, H.S. Eden, J.M. Farber, and X. Chen, Improvement of CXCR4 tracer specificity for PET imaging. J Control Release, 2011. [PMC free article: PMC3259211] [PubMed: 21964282]


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