Radiotherapy can facilitate the immune recognition of immunologically “cold” tumors and enhance the efficacy of anti-PD-1 and anti-CTLA-4 immune checkpoint inhibitors (ICIs) in melanoma. Systemic administration of receptor-targeted radionuclide therapy has the potential to selectively deliver radionuclides to multiple tumors throughout the body in metastatic settings. By triggering immunologic cell death and increasing the immune susceptibility of surviving tumor cells in these locations, targeted radionuclide therapies may overcome resistance to ICIs and render immunologically “cold” tumors throughout the body responsive to ICIs and immunologically “hot”. Here, we show the anti-tumor cooperation of targeted α-particle radionuclide therapy (α-TRT) and ICIs in preclinical models of melanoma. Melanocortin 1 receptor (MC1R)-targeted radiopeptide [²¹²Pb]VMT01 was employed to deliver α-radiation to melanoma tumors in mice. A single injection of 4.1 MBq [²¹²Pb]VMT01 significantly slowed the tumor growth of B16-F10 melanoma and the combination of [²¹²Pb]VMT01 and ICIs induced a cooperative anti-tumor effect leading to 43% complete tumor response with no sign of malignancy on autopsy. Animals with complete response developed anti-tumor immunity to reject further tumor inoculations. This therapeutic cooperation was completely abolished in RAG1 KO mice, which are deficient in T-cell maturation. In addition, the anti-tumor cooperation was compromised when fractionated [²¹²Pb]VMT01 was used in the combination. We also demonstrated that [²¹²Pb]VMT01 induced immunogenic cell death in tumor vaccination assays and in vitro exposure to [²¹²Pb]VMT01 sensitized immunotolerant melanoma to ICIs treatment in vivo. Enhanced tumor infiltrating CD3⁺, CD4⁺, CD8⁺ lymphocytes were observed following injection of 1.4 MBq [²¹²Pb]VMT01. Overall, we demonstrated anti-tumor cooperation between α-TRT and ICIs in melanoma that is mediated by tumor specific immunity.

Receptor-targeted image-guided Radionuclide Therapy (TRT) is increasingly recognized as a promising approach to cancer treatment. In particular, the potential for clinical translation of receptor-targeted alpha-particle therapy is receiving considerable attention as an approach that can improve outcomes for cancer patients. Higher Linear-energy Transfer (LET) of alpha-particles (compared to beta particles) for this purpose results in an increased incidence of double-strand DNA breaks and improved-localized cancer-cell damage. Recent clinical studies provide compelling evidence that alpha-TRT has the potential to deliver a significantly more potent anti-cancer effect compared with beta-TRT. Generator-produced 212Pb (which decays to alpha emitters 212Bi and 212Po) is a particularly promising radionuclide for receptor-targeted alpha-particle therapy. A second attractive feature that distinguishes 212Pb alpha-TRT from other available radionuclides is the possibility to employ elementallymatched isotope 203Pb as an imaging surrogate in place of the therapeutic radionuclide. As direct non-invasive measurement of alpha-particle emissions cannot be conducted using current medical scanner technology, the imaging surrogate allows for a pharmacologically-inactive determination of the pharmacokinetics and biodistribution of TRT candidate ligands in advance of treatment. Thus, elementally-matched 203Pb labeled radiopharmaceuticals can be used to identify patients who may benefit from 212Pb alpha-TRT and apply appropriate dosimetry and treatment planning in advance of the therapy. In this review, we provide a brief history on the use of these isotopes for cancer therapy; describe the decay and chemical characteristics of 203/212Pb for their use in cancer theranostics and methodologies applied for production and purification of these isotopes for radiopharmaceutical production. In addition, a medical physics and dosimetry perspective is provided that highlights the potential of 212Pb for alpha-TRT and the expected safety for 203Pb surrogate imaging. Recent and current preclinical and clinical studies are presented. The sum of the findings herein and observations presented provide evidence that the 203Pb/212Pb theranostic pair has a promising future for use in radiopharmaceutical theranostic therapies for cancer.

²⁰³Pb and ²¹²Pb have emerged as promising theranostic isotopes for image-guided α-particle radionuclide therapy for cancers. Here, we report a cyclen-based Pb specific chelator (PSC) that is conjugated to tyr³-octreotide via a PEG₂ linker (PSC-PEG-T) targeting somatostatin receptor subtype 2 (SSTR2). PSC-PEG-T could be labeled efficiently to purified ²¹²Pb at 25 °C and also to ²¹²Bi at 80 °C. Efficient radiolabeling of mixed ²¹²Pb and ²¹²Bi in PSC-PEG-T was also observed at 80 °C. Post radiolabeling, stable Pb(II) and Bi(III) radiometal complexes in saline were observed after incubating [²⁰³Pb]Pb-PSC-PEG-T for 72 h and [²¹²Bi]Bi-PSC-PEG-T for 5 h. Stable [²¹²Pb]Pb-PSC-PEG-T and progeny [²¹²Bi]Bi-PSC-PEG-T were identified after storage in saline for 24 h. In serum, stable radiometal/radiopeptide were observed after incubating [²⁰³Pb]Pb-PSC-PEG-T for 55 h and [²¹²Pb]Pb-PSC-PEG-T for 24 h. In vivo biodistribution of [²¹²Pb]Pb-PSC-PEG-T in tumor-free CD-1 Elite mice and athymic mice bearing AR42J xenografts revealed rapid tumor accumulation, excellent tumor retention and fast renal clearance of both ²¹²Pb and ²¹²Bi, with no in vivo redistribution of progeny ²¹²Bi. Single-photon emission computed tomography (SPECT) imaging of [²⁰³Pb]Pb-PSC-PEG-T and [²¹²Pb]Pb-PSC-PEG-T in mice also demonstrated comparable accumulation in AR42J xenografts and renal clearance, confirming the theranostic potential of the elementally identical ²⁰³Pb/²¹²Pb radionuclide pair.

[²¹²Pb]VMT01 is a melanocortin 1 receptor (MC1R) targeted theranostic ligand in clinical development for alpha particle therapy for melanoma. ²¹²Pb has an elementally matched gamma-emitting isotope ²⁰³Pb; thus, [²⁰³Pb]VMT01 can be used as an imaging surrogate for [²¹²Pb]VMT01. [²¹²Pb]VMT01 human serum stability studies have demonstrated retention of the ²¹²Bi daughter within the chelator following beta emission of parent ²¹²Pb. However, the subsequent alpha emission from the decay of ²¹²Bi into ²⁰⁸Tl results in the generation of free ²⁰⁸Tl. Due to the 10.64-hour half-life of ²¹²Pb, accumulation of free ²⁰⁸Tl in the injectate will occur. The goal of this work is to estimate the human dosimetry for [²¹²Pb]VMT01 and the impact of free ²⁰⁸Tl in the injectate on human tissue absorbed doses. Human [²¹²Pb]VMT01 tissue absorbed doses were estimated from murine [²⁰³Pb]VMT01 biodistribution data, and human biodistribution values for ²⁰¹Tl chloride (a cardiac imaging agent) from published data were used to estimate the dosimetry of free ²⁰⁸Tl. Results indicate that the dose-limiting tissues for [²¹²Pb]VMT01 are the red marrow and the kidneys, with estimated absorbed doses of 1.06 and 8.27 mGy_{RBE = 5}/MBq. The estimated percent increase in absorbed doses from free ²⁰⁸Tl in the injectate is 0.03% and 0.09% to the red marrow and the kidneys, respectively. Absorbed doses from free ²⁰⁸Tl result in a percent increase of no more than 1.2% over [²¹²Pb]VMT01 in any organ or tissue. This latter finding indicates that free ²⁰⁸Tl in the [²¹²Pb]VMT01 injectate will not substantially impact estimated tissue absorbed doses in humans.