The validation study of a new fluorescence-target tracer for sentinel lymph node biopsy

丛斌斌; 刘治国; 孙　晓; 曹晓珊; 邱鹏飞; 王春建; 张朝蓬; 田崇麟; 吴　爽; 王永胜

doi:10.19401/j.cnki.1007-3639.2020.03.004

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The validation study of a new fluorescence-target tracer for sentinel lymph node biopsy

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- The validation study of a new fluorescence-target tracer for sentinel lymph node biopsy
- China Oncology Vol. 30, Issue 3, Pages: 179-185(2020)
- 作者机构：
  
  1. 山东省肿瘤防治研究院（山东省肿瘤医院）乳腺病中心,山东,济南,250117
  2. 山东第一医科大学（山东省医学科学院）,山东,济南,250062
  3. 山东省肿瘤防治研究院（山东省肿瘤医院）核医学科,山东,济南,250117
- 作者简介：
- 基金信息：
- DOI：10.19401/j.cnki.1007-3639.2020.03.004
  CLC： R737.9
- Published Online：03 April 2020，
  
  Published：03 April 2020
- 稿件说明：
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丛斌斌 , 刘治国 , 孙　晓 , 曹晓珊 , 邱鹏飞 , 王春建 , 张朝蓬 , 田崇麟 , 吴　爽 , 王永胜 . 新型荧光靶向前哨淋巴结示踪剂的验证研究[J]. 中国癌症杂志, 2020, 30(3): 179-185.

丛斌斌, 刘治国, 孙　晓, et al. The validation study of a new fluorescence-target tracer for sentinel lymph node biopsy[J]. China Oncology, 2020, 30(3): 179-185.
丛斌斌 , 刘治国 , 孙　晓 , 曹晓珊 , 邱鹏飞 , 王春建 , 张朝蓬 , 田崇麟 , 吴　爽 , 王永胜 . 新型荧光靶向前哨淋巴结示踪剂的验证研究[J]. 中国癌症杂志, 2020, 30(3): 179-185. DOI： 10.19401/j.cnki.1007-3639.2020.03.004.

丛斌斌, 刘治国, 孙　晓, et al. The validation study of a new fluorescence-target tracer for sentinel lymph node biopsy[J]. China Oncology, 2020, 30(3): 179-185. DOI： 10.19401/j.cnki.1007-3639.2020.03.004.

摘要

背景与目的：示踪剂是前哨淋巴结活检的关键，传统荧光示踪剂无法靶向前哨淋巴结且荧光强度弱。探寻一种近红外荧光强度更大、穿透力更好的新型增强近红外荧光示踪剂聚甲川菁染料分子Cy754，并将其与利妥昔单抗（rituximab，Rit）结合形成新型近红外荧光靶向示踪剂Cy754-Rit。方法：化学合成新型增强近红外荧光示踪剂Cy754，检测其相对分子质量、激发光谱、吸收光谱等，将其与Rit进行直接偶联。测定Cy754-Rit标记率，检测新型示踪剂中单抗分子完整性和免疫活性。采用近红外成像仪将Cy754-Rit与吲哚菁绿（indocyanine green，ICG）-Rit的荧光强度作比较，检测其安全限度，通过前哨淋巴结动物模型验证其前哨淋巴结的定位特性，并与Cy754、ICG-Rit、核素进行对比。结果：Cy754的相对分子质量为818，激发波长为740 nm，发射波长为760 nm。直接偶联并纯化的Cy754-Rit保持了分子完整性和免疫活性。Rit上的Cy754标记率为100%。Cy754-Rit的荧光强度优于ICG-Rit（3.08×10

vs 6.56×10

）。透析纯化后的新型增强靶向示踪剂中未检测到细菌和热原。实验动物局部注射，观察2周后，局部及全身无红肿也无皮疹，无死亡。Rit与Cy754偶联的最佳的物质的量比为1∶80。Cy754-Rit能够准确地定位前哨淋巴结，其定位特性与核素相似，但发射荧光穿透力优于ICG-Rit（14.3 mm vs 13.2 mm）。结论：Cy754-Rit的制备工艺简便、高效、无放射性，其荧光强度较传统荧光靶向示踪剂明显增强。经动物模型局部注射能够准确地定位到前哨淋巴结，显像清晰稳定，穿透力好，但仍需临床试验进行相关验证。

Abstract

Background and purpose: Tracer is the key to sentinel lymph node biopsy

but the traditional fluorescence tracer could not target at sentinel lymph node with weak intensity. A new near-infrared fluorescence tracer polymethylene cyani

ne dye Cy754 with stronger permeability and intensity was synthetized and combined with rituximab (Rit) to produce a new near-infrared fluorescence-target tracer Cy754-Rit. Methods: The new near-infrared fluorescence tracer Cy754 was synthetized by chemical method

and the basic parameters including molecular weight

absorption and emission spectra were tested. The molecule of Cy754 was combined directly with Rit to produce Cy754-Rit. The Cy754-Rit was analyzed for labeled rate

molecular integrity

and molecular immune activity. The fluorescence intensity of Cy754-Rit was compared with indocyanine green (ICG)-Rit by using near-infrared fluorescence detector. The safety limitation was tested. The localization ability of Cy754-Rit was tested in animal model of sentinel lymph node and compared with Cy754

ICG-Rit and radiotracer. Results: The molecular weight of Cy754 was 818

and the absorption and emission spectra were 740 nm and 760 nm. The Cy754-Rit was intact and kept the immune activity of Rit. The Cy754 labeled rate of Rit was 100%. The fluorescence intensity of Cy754-Rit was stronger than that of ICG-Rit (3.08×10

vs 6.56×10

). The Cy754-Rit was bacteria- and pyrogen-free

and was safe to body with location injection. The best molarity ratio of Cy754-Rit was 80∶1. The Cy754-Rit could identify the location of sentinel lymph node which was accorded with the radiotracer and had a stronger permeability than ICG-Rit (14.3 mm vs 13.2 mm). Conclusion: The method of producing Cy754-Rit is simple

effective and non-radioactive. The fluorescence intensity of Cy754-Rit is stronger than that of traditional tracer. The Cy754-Rit could identify the location of sentinel lymph node in the animal model of sentinel lymph node but should be validated by clinical trial before its use in clinical practice.

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