<sup>68</sup>Ga-DOTA-PDL1P的设计合成及其在黑色素瘤小鼠模型中的应用研究

杨梓怡; 李盼丽; 顾丙新; 刘成; 宋少莉; 许晓平

doi:10.19401/j.cnki.1007-3639.2023.04.005

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⁶⁸Ga-DOTA-PDL1P的设计合成及其在黑色素瘤小鼠模型中的应用研究

论著 | 更新时间：2025-12-31

- ⁶⁸Ga-DOTA-PDL1P的设计合成及其在黑色素瘤小鼠模型中的应用研究
- The synthesis of a novel molecular imaging probe ⁶⁸Ga-DOTA-PDL1P and application in mouse model of melanoma
- 中国癌症杂志 2023年33卷第4期页码：354-360
- 作者机构：
  
  1. 复旦大学附属肿瘤医院核医学科，复旦大学上海医学院肿瘤学系，上海 200032
  2. 复旦大学生物医学影像研究中心，上海 200032
  3. 上海分子影像探针工程技术中心，上海 200032
  4. 上海市质子重离子医院核医学科，上海 201315
  5. 核物理与离子束应用教育部重点实验室，上海 200433
- 作者简介：
  
  [ "杨梓怡（ORCID: 0000-0002-8947-2623），住院医师。" ]
  许晓平（ORCID: 0000-0001-9656-6233），药师，副研究员。
- 基金信息：
  
  国家自然科学基金(12275057);国家自然科学基金(81971648);国家自然科学基金重点项目(32030061);专利(CN 110981940 A)
- DOI：10.19401/j.cnki.1007-3639.2023.04.005
  中图分类号： R730.44
- 收稿：2022-09-05，
  
  修回：2023-03-08，
  
  纸质出版：2023-04-30
- 稿件说明：
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杨梓怡, 李盼丽, 顾丙新, 等. ⁶⁸Ga-DOTA-PDL1P的设计合成及其在黑色素瘤小鼠模型中的应用研究[J]. 中国癌症杂志, 2023,33(4):354-360.

Ziyi YANG, Panli LI, Bingxin GU, et al. The synthesis of a novel molecular imaging probe ⁶⁸Ga-DOTA-PDL1P and application in mouse model of melanoma[J]. China Oncology, 2023, 33(4): 354-360.
杨梓怡, 李盼丽, 顾丙新, 等. ⁶⁸Ga-DOTA-PDL1P的设计合成及其在黑色素瘤小鼠模型中的应用研究[J]. 中国癌症杂志, 2023,33(4):354-360. DOI： 10.19401/j.cnki.1007-3639.2023.04.005.

Ziyi YANG, Panli LI, Bingxin GU, et al. The synthesis of a novel molecular imaging probe ⁶⁸Ga-DOTA-PDL1P and application in mouse model of melanoma[J]. China Oncology, 2023, 33(4): 354-360. DOI： 10.19401/j.cnki.1007-3639.2023.04.005.

摘要

背景与目的：

以程序性死亡[蛋白

]

-1（programmed death-1，PD-1）/程序性死亡[蛋白

]

配体-1（programmed death ligand-1，PD-L1）抑制剂为代表的免疫治疗药物在临床上获得巨大成功，但整体有效率差异很大。现阶段在肿瘤开始治疗前，检测患者PD-1/PD-L1的表达情况是临床用药的重要依据，但目前检测PD-L1表达的方法需要获取患者的标本，而获取标本存在创伤性。因此，迫切需要开发一种无创性活体内特异性检测PD-L1表达的方法。我们设计合成了一种新型靶向PD-L1多肽的正电子发射断层显像（positron emission tomography，PET）显像剂

Ga-DOTA-PDL1P，本研究通过对

Ga-DOTA-PDL1P的标记率、放射化学纯度及稳定性进行质量控制评估，并在黑色素瘤小鼠模型中进行应用评价，以期获得有转化前景的

Ga-DOTA-PDL1P新型免疫PET探针。

方法：

利用镓-68（

Ga）标记 DOTA-PDL1P，并对其放射性化学纯度和稳定性进行质量控制评估；使用鼠源性黑色素瘤B16-F10细胞系进行体外细胞摄取及阻断实验以评价其特异靶向性能；采用B16-F10荷瘤小鼠进行生物分布实验分析该PET显像剂在体内的代谢情况；通过PET/CT显像观察

Ga-DOTA-PDL1P 在B16-F10荷瘤小鼠中的肿瘤摄取程度并进行半定量分析肿瘤靶本比（tumor/muscle，T/M）；另外对肿瘤样本进行放射自显影进一步分析肿瘤对

Ga-DOTA-PDL1P探针的摄取情况。采用肿瘤细胞阳性比例分数（tumor cell proportion score，TPS）及联合阳性分数（combined positive score，CPS）指标分析免疫组织化学染色结果评价肿瘤组织的PD-L1表达情况。

结果：

Ga-DOTA-PDL1P的标记率及放射化学纯度均大于99%，并在磷酸缓冲盐溶液（phosphate-buffered saline，PBS）和胎牛血清（fetal bovine serum，FBS）中温育3 h后仍保持较好的稳定性（放化纯度均在90%以上）。细胞摄取结果示，

Ga-DOTA-PDL1P可被B16-F10细胞特异性摄取，且该特异性摄取能够被PD-L1P多肽竞争结合阻断。生物分布实验结果示

Ga-DOTA-PDL1P主要经泌尿系统排泄。荷瘤小鼠microPET/CT显像结果表明肿瘤组织高度摄取

Ga-DOTA-PDL1P，T/M为6.7。放射自显影结果证实肿瘤

Ga-DOTA-PDL1P高摄取部位与免疫组织化学染色PD-L1表达阳性区域一致，证明

Ga-DOTA-PDL1P能够特异性与PD-L1阳性的肿瘤细胞结合。

结论：

本研究合成的

Ga-DOTA-PDL1P新型PET显像剂稳定性好，能特异性靶向PD-L1阳性肿瘤细胞，是一种有转化前景的靶向PD-L1蛋白的新型免疫PET探针。

Abstract

Background and purpose:

The immunogenic drugs represented by programmed cell death 1 (PD-1) and programmed death ligand-1 (PD-L1) have achieved great success in clinic. However

with the development of clinical application

the overall effective rate varies greatly. At present

detection of the expression of PD-1/PD-L1 before the start of tumor treatme

nt is an important basis for clinical medication. However

the current detection method of PD-L1 expression requires the acquisition of patient specimens

which is traumatic. Therefore

there is an urgent need to develop a noninvasive method to detect PD-L1 expression

in vivo

. This study innovatively designed and synthesized a new positron emission tomography (PET) imaging agent targeting PD-L1 polypeptide

Ga-DOTA-PDL1P. In this study

the labeling rate

radiochemical purity and stability of

Ga-DOTA-PDL1P were evaluated for quality control. And the application was evaluated in the mouse model of melanoma in order to obtain a promising

Ga-DOTA-PDL1P immune PET probe.

Methods:

Gallium-68 (

Ga) was used to label DOTA-PDL1P

and its radiochemical purity and stability were evaluated by quality control. To evaluate its specific targeting performance

the cell uptake and blocking experiments were carried out in murine melanoma B16-F10 cells. The biodistribution experiment was performed in B16-F10 tumor-bearing mice to analyze the metabolism of the PET imaging agent

in vivo

. The Tumor uptake of

Ga-DOTA-PDL1P in B16-F10 tumor-bearing mice was observed by PET/CT imaging

and the tumor/muscle (T/M) ratio was semi-quantitatively analyzed. In addition

radioautography was performed to further analyze the tumor uptake of

Ga-DOTA-PDL1P probe. Tumor cell proportion score (TPS) and combined positive score (CPS) were used to analyze the immunohistochemical staining results to evaluate the expression of PD-L1 in tumor tissues.

Results:

The labeling efficiency and radiochemical purity of

Ga-DOTA-PDL1P were more than 99%

and the radiochemical purity of

Ga-DOTA-PDL1P was more than 90% after 3 h of incubation in PBS and fetal bovine serum. Cellular uptake results showed that

Ga-DOTA-PDL1P could be specifically taken up by B16-F10 cells

and the specific uptake c

ould be blocked by PD-L1P peptide competitive binding. Biodistribution assay showed that

Ga-DOTA-PDL1P was mainly excreted through the urinary system. The results of microPET/CT imaging of tumor-bearing mice showed a high uptake of

Ga-DOTA-PDL1P in tumor tissues

and the T/M ratio was 6.7. The autoradiography results confirmed that

Ga-DOTA-PDL1P high uptake in the tumor was consistent with the PD-L1 positive area of immunohistochemical staining

which proved that

Ga-DOTA-PDL1P could specifically bind to PD-L1 positive malignant tumor cells.

Conclusion:

The novel

Ga-DOTA-PDL1P PET imaging agent synthesized in this study has good stability and can specifically target PD-L1 positive tumor cells

which is a promising new immune PET probe targeting PD-L1 protein.

关键词

Keywords

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68Ga-DOTA-PDL1P的设计合成及其在黑色素瘤小鼠模型中的应用研究

The synthesis of a novel molecular imaging probe 68Ga-DOTA-PDL1P and application in mouse model of melanoma

DOI：10.19401/j.cnki.1007-3639.2023.04.005

摘要

Abstract

关键词

Keywords

references

⁶⁸Ga-DOTA-PDL1P的设计合成及其在黑色素瘤小鼠模型中的应用研究

The synthesis of a novel molecular imaging probe ⁶⁸Ga-DOTA-PDL1P and application in mouse model of melanoma