Preliminary evaluation of Lu-177-labeled knottin peptides for integrin receptor-targeted radionuclide therapy

Department of Nuclear Medicine, Shanghai Ruijin Hospital, Shanghai Jiaotong University, 2nd Ruijin Rd, Shanghai, 200025, People's Republic of China.
European Journal of Nuclear Medicine (Impact Factor: 5.38). 12/2010; 38(4):613-22. DOI: 10.1007/s00259-010-1684-x
Source: PubMed


Cystine knot peptides (knottins) 2.5D and 2.5F were recently engineered to bind integrin receptors with high affinity and specificity. These receptors are overexpressed on the surface of a variety of malignant human tumor cells and tumor neovasculature. In this study, 2.5D and 2.5F were labeled with a therapeutic radionuclide, (177)Lu, and the resulting radiopeptides were then evaluated as potential radiotherapeutic agents in a murine model of human glioma xenografts.
Knottins 2.5D and 2.5F were synthesized using solid phase peptide synthesis, folded in vitro, and site-specifically coupled with 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA) at their N terminus for (177)Lu radiolabeling. The stability of the radiopeptides (177)Lu-DOTA-2.5D and (177)Lu-DOTA-2.5F was tested in both phosphate-buffered saline (PBS) and mouse serum. Cell uptake assays of the radiolabeled peptides were performed in U87MG integrin-expressing human glioma cells. The biodistribution studies of both (177)Lu-DOTA-2.5D and (177)Lu-DOTA-2.5F were examined in U87MG tumor-bearing athymic nu/nu mice. Radiation absorbed doses for the major tissues of a human adult male were calculated based on the mouse biodistribution results.
DOTA-2.5D and DOTA-2.5F were labeled with (177)Lu at over 55% efficiency. High radiochemical purity for both radiocomplexes (> 95%) could be achieved after high performance liquid chromatography (HPLC) purification. Both radiopeptides were stable in PBS and mouse serum. Compared to (177)Lu-DOTA-2.5D (0.39 and 0.26 %ID/g at 2 and 24 h, respectively), (177)Lu-DOTA-2.5F showed much higher tumor uptake (2.16 and 0.78 %ID/g at 2 and 24 h, respectively). It also displayed higher tumor to blood ratios than that of (177)Lu-DOTA-2.5D (31.8 vs 18.7 at 24 h and 52.6 vs 20.6 at 72 h). Calculation of radiodosimetry for (177)Lu-DOTA-2.5D and (177)Lu-DOTA-2.5F suggested that tumor and kidney were tissues with the highest radiation absorbed doses. Moreover, (177)Lu-DOTA-2.5F had a higher tumor to kidney radiation absorbed dose ratio than that of (177)Lu-DOTA-2.5D.
Cystine knot peptides can be successfully radiolabeled with (177)Lu for potential therapeutic applications. Knottin 2.5F labeled with (177)Lu exhibits favorable distribution in murine U87MG xenograft model; thus, it is a promising agent for radionuclide therapy of integrin-positive tumors.

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    • "The power of theranostics is already established in the clinic where 68 Ga/ 177 Lu-labeled peptides targeting the somatostain receptor are successfully being used for imaging and treatment of neuroendocrine tumors [18]. Several new targets are currently being evaluated in pre-clinical cancer models including integrins [19], HER-2 [20] [21], gastrin-releasing peptide (GRP) [22] and vascular endothelial growth factor (VEGF) [23]. 177 Lu is a low-energy betaemitter (~1.5 mm maximum penetration in soft tissue) capable of inducing cytotoxic effects in tumors but not surrounding tissue by providing both a 'cross-fire'-and 'bystander' effect from direct betaparticles and Auger electrons, respectively [24]. "
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    • "But due to the lower tumor uptake of the RGD monomer, multiple-dose administration was necessary to achieve therapeutic efficacy [106]. Recently, 177Lu labeled two knottin peptides (2.5D and 2.5F), and RGD peptides targeting a range of integrins (αvβ3/αvβ5/α5β) were tested for potential radiotherapy in a mouse model of human glioma [107, 108]. 177Lu-DOTA-2.5F "
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    05/2012; 2012(12):817682. DOI:10.1155/2012/817682
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    • "Several knottin mutants that bind to integrin receptors (αvβ3/αvβ5 or αvβ3/αvβ5/α5β1) with low nanomolar affinity have been identified 51, 52, and radionuclide and optical dye labeled such knottin peptides have demonstrated favorable in vivo tumor targeting properties 53-56. Recently, two knottin peptides (2.5D and 2.5F: targeting integrin αvβ3/αvβ5 and αvβ3/αvβ5/α5β1, respectively) were radiolabeled with a therapeutic radionuclide 177Lu, and the resulting radiopharmaceuticals were evaluated for potential radiotherapy in a mouse model of human glioma 57. Compared to 177Lu-DOTA-2.5D, "
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