Preclinical Evaluation of⁶⁸Ga- and¹⁷⁷Lu-Labeled Integrin α_vβ₆-Targeting Radiotheranostic Peptides

Abstract

The integrin α_vβ₆, an epithelium-specific cell surface receptor, is overexpressed on numerous malignancies, including the highly lethal pancreatic ductal adenocarcinomas. Here, we developed and tested a novel α_vβ₆-targeting peptide, DOTA-5G (<b>1</b>) radiolabeled with ⁶⁸Ga, for PET/CT imaging and ¹⁷⁷Lu for treatment. With the goal to develop a radiotheranostic, further modifications were made for increased circulation time, renal recycling, and tumor uptake, yielding DOTA-albumin-binding moiety-5G (<b>2</b>). <b>Methods:</b> Peptides <b>1</b> and <b>2</b> were synthesized on solid phase, and their affinity for α_vβ₆ was assessed by enzyme-linked immunosorbent assay. The peptides were radiolabeled with ⁶⁸Ga and ¹⁷⁷Lu. In vitro cell binding, internalization, and efflux of ⁶⁸Ga-<b>1</b> and ¹⁷⁷Lu-<b>2</b> were evaluated in α_vβ₆-positive BxPC-3 human pancreatic cancer cells. PET/CT imaging of ⁶⁸Ga-<b>1</b> and ⁶⁸Ga-<b>2</b> was performed on female nu/nu mice bearing subcutaneous BxPC-3 tumors. Biodistribution was performed for ⁶⁸Ga-<b>1</b> (1 and 2 h after injection), ⁶⁸Ga-<b>2</b> (2 and 4 h after injection), and ¹⁷⁷Lu-<b>1</b> and ¹⁷⁷Lu-<b>2</b> (1, 24, 48, and 72 h after injection). The ¹⁷⁷Lu-<b>2</b> biodistribution data were extrapolated for human dosimetry data estimates using OLINDA/EXM 1.1. Therapeutic efficacy of ¹⁷⁷Lu-<b>2</b> was evaluated in mice bearing BxPC-3 tumors. <b>Results:</b> Peptides <b>1</b> and <b>2</b> demonstrated high affinity (&lt;55 nM) for α_vβ₆ by enzyme-linked immunosorbent assay. ⁶⁸Ga-<b>1</b>, ⁶⁸Ga-<b>2</b>, ¹⁷⁷Lu-<b>1</b>, and ¹⁷⁷Lu-<b>2</b> were synthesized in high radiochemical purity. Rapid in&nbsp;vitro binding and internalization of ⁶⁸Ga-<b>1</b> and ¹⁷⁷Lu-<b>2</b> were observed in BxPC-3 cells. PET/CT imaging and biodistribution studies demonstrated uptake in BxPC-3 tumors. Introduction of the albumin-binding moiety in ¹⁷⁷Lu-<b>2</b> resulted in a 5-fold increase in tumor uptake and retention over time. Based on the extended dosimetry data, the dose-limiting organ for ¹⁷⁷Lu-<b>2</b> is the kidney. Treatment with ¹⁷⁷Lu-<b>2</b> prolonged median survival by 1.5- to 2-fold versus controls. <b>Conclusion:</b>⁶⁸Ga-<b>1</b> and ¹⁷⁷Lu-<b>2</b> demonstrated high affinity for the integrin α_vβ₆ both in&nbsp;vitro and in&nbsp;vivo, were rapidly internalized into BxPC-3 cells, and were stable in mouse and human serum. Both radiotracers showed favorable pharmacokinetics in preclinical studies, with predominantly renal excretion and good tumor–to–normal-tissue ratios. Favorable human dosimetry data suggest the potential of ¹⁷⁷Lu-<b>2</b> as a treatment for pancreatic ductal adenocarcinoma.

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