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Development of small-molecular-based radiotracers for PET imaging of PD-L1 expression and guiding the PD-L1 therapeutics

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European Journal of Nuclear Medicine and Molecular Imaging Aims and scope Submit manuscript

Abstract

Purpose

Programmed cell death protein ligand 1 (PD-L1) is a crucial biomarker for immunotherapy. However, nearly 70% of patients do not respond to PD-L1 immune checkpoint therapy. Accurate monitoring of PD-L1 expression and quantification of target binding during treatment are essential. In this study, a series of small-molecule radiotracers were developed to assess PD-L1 expression and direct immunotherapy.

Methods

Radiotracers of [68Ga]Ga-D-PMED, [68Ga]Ga-D-PEG-PMED, and [68Ga]Ga-D-pep-PMED were designed based on a 2-methyl-3-biphenyl methanol scaffold and successfully synthesized. Cellular experiments and molecular docking assays were performed to determine their specificity for PD-L1. PD-L1 status was investigated via positron emission tomography (PET) imaging in MC38 tumor models. PET imaging of [68Ga]Ga-D-pep-PMED was performed to noninvasively quantify PD-L1 blocking using an anti-mouse PD-L1 antibody (PD-L1 mAb).

Results

The radiosyntheses of [68Ga]Ga-D-PMED, [68Ga]Ga-D-PEG-PMED, and [68Ga]Ga-D-pep-PMED were achieved with radiochemical yields of 87 ± 6%, 82 ± 4%, and 79 ± 9%, respectively. In vitro competition assays demonstrated their high affinities (the IC50 values of [68Ga]Ga-D-PMED, [68Ga]Ga-D-PEG-PMED, and [68Ga]Ga-D-pep-PMED were 90.66 ± 1.24, 160.8 ± 1.35, and 51.6 ± 1.32 nM, respectively). At 120 min postinjection (p.i.) of the radiotracers, MC38 tumors displayed optimized tumor-to-muscle ratios for all radioligands. Owing to its hydrophilic modification, [68Ga]Ga-D-pep-PMED had the highest target-to-nontarget (T/NT) ratio of approximately 6.2 ± 1.2. Interestingly, the tumor/liver ratio was hardly affected by different concentrations of the inhibitor BMS202. We then evaluated the impacts of dose and time on accessible PD-L1 levels in the tumor during anti-mouse PD-L1 antibody treatment. The tumor uptake of [68Ga]Ga-D-pep-PMED significantly decreased with increasing PD-L1 mAb dose. Moreover, after 8 days of treatment with a single antibody, the uptake of [68Ga]Ga-D-pep-PMED in the tumor significantly increased but remained lower than that in the saline group.

Conclusion

PET imaging with [68Ga]Ga-D-pep-PMED, a small-molecule radiotracer, is a promising tool for evaluating PD-L1 expression and quantifying the target blockade of PD-L1 to assist in the development of effective therapeutic regimens.

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Data availability

All data relevant to the study are included in the article or uploaded as supplementary information.

Abbreviations

ICIs:

Immune checkpoint inhibitors

ICB:

Immune checkpoint blockade

PD-1:

Programmed cell death protein-1

PD-L1:

Programmed death ligand-1

IHC:

Immunohistochemistry

PET:

Positron emission tomography

mAbs:

Monoclonal antibodies

TACs:

Time-activity curves

NCCN:

National Comprehensive Cancer Network

HPLC:

High-performance liquid chromatography

ROI:

Regions of interest

RCY:

Radiochemical yield

T/NT:

Target-to-nontarget

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Funding

This study was financially supported by the National Natural Science Foundation of China (82372008, 81901805, 21976150), Joint Fund of the National Natural Science Foundation of China—China National Nuclear Corporation for Nuclear Technology Innovation (U1967222), and Shanghai Pujiang Program (22PJ1401700).

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Authors and Affiliations

  1. State Key Laboratory of Vaccines for Infectious Diseases, Center for Molecular Imaging and Translational Medicine, Xiang An Biomedicine Laboratory, School of Public Health, Xiamen University, Xiamen, 361102, China

    Hongzhang Yang, Xinying Zeng, Jia Liu, Xuejun Wen, Huanhuan Liu, Yuanyuan Liang, Xueqi Wang, Jianyang Fang, Qinglin Zhang & Zhide Guo

  2. Department of Nuclear Medicine, Fudan University Shanghai Cancer Center, Shanghai, 200032, China

    Jindian Li

  3. Theranostics and Translational Research Center, Institute of Clinical Medicine & Department of Nuclear Medicine, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, No. 1 Shuaifuyuan, Beijing, 100730, China

    Xianzhong Zhang

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  1. Hongzhang Yang
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Contributions

HY, XZ, and JL: conceptualization, data curation, formal analysis, validation, investigation, visualization, methodology, writing—original draft and editing. XW, HL, YL, XW, JF, and QZ: investigation, methodology. JL, XZ, and ZG: conceptualization, resources, supervision, funding acquisition, investigation, writing—review and editing.

Corresponding authors

Correspondence to Jindian Li, Xianzhong Zhang or Zhide Guo.

Ethics declarations

Ethics approval

All animal studies were approved by the Institutional Animal Care and Use Committee of Laboratory Animals Center for Xiamen University. And the protocols were prepared before the study.

Conflict of interest

The authors declare no competing interests.

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Hongzhang Yang, Xinying Zeng, and Jia Liu were the first authors and contributed equally to this work.

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Yang, H., Zeng, X., Liu, J. et al. Development of small-molecular-based radiotracers for PET imaging of PD-L1 expression and guiding the PD-L1 therapeutics. Eur J Nucl Med Mol Imaging 51, 1582–1592 (2024). https://doi.org/10.1007/s00259-024-06610-3

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