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GnRH (Gonadotropin-releasing hormone) receptor-targeted near-infrared fluorescence imaging probe and preparation method and application thereof

A fluorescent imaging and near-infrared technology, applied in the biological field, can solve the problem of high false positive rate in ovarian cancer lesion imaging, achieve deep penetration depth and good application prospects

Active Publication Date: 2019-08-02
THE OBSTETRICS & GYNECOLOGY HOSPITAL OF FUDAN UNIV
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Problems solved by technology

However, ICG cannot specifically bind to tumor cells, and the false positive rate of direct imaging of ovarian cancer lesions is as high as 62%.

Method used

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  • GnRH (Gonadotropin-releasing hormone) receptor-targeted near-infrared fluorescence imaging probe and preparation method and application thereof
  • GnRH (Gonadotropin-releasing hormone) receptor-targeted near-infrared fluorescence imaging probe and preparation method and application thereof
  • GnRH (Gonadotropin-releasing hormone) receptor-targeted near-infrared fluorescence imaging probe and preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment

[0036] The specific implementation of the above-mentioned near-infrared fluorescent probe in tumor targeted imaging application is as follows:

[0037] Inoculate human ovarian cancer cell line A2780 (high GnRH receptor expression) and human lung cancer cell line H1299 (low GnRH receptor expression) in the logarithmic growth phase on a chamber slide, and start the experiment when the cell confluence reaches 50% . Aspirate the culture medium and wash once with PBS buffer. 60μM GnRHa-ICG and ICG were added, and incubated with tumor cells at 37°C for 30 minutes. Aspirate the supernatant and wash 3 times with PBS. Add 4% paraformaldehyde and fix at room temperature for 10 min. Aspirate the fixative and wash with PBS once. Add cell membrane dye WGA488 and incubate at room temperature for 10 min. Absorb the membrane dye and wash with PBS 3 times. Mount the slide with a fluorescent mounting tablet containing DAPI, and observe the fluorescence intensity with a confocal microscope.

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Abstract

The invention provides a GnRH (Gonadotropin-releasing hormone) receptor-targeted near-infrared fluorescent imaging probe GnRHa-ICG and a preparation method and application thereof. The GnRH receptor-targeted near-infrared fluorescent imaging probe GnRHa-ICGis formed by coupling a GnRH polypeptide (sequence D-2-Nal-D-4-Cl-Phe-D-3-Pal-Ser-Tyr-D-Cit-Leu-Arg-Pro-D-Ala-NH2) and activated near-infraredfluorescent dye ICG-NHS. Based on the principle of specific binding of the GnRH polypeptide to the GnRH receptor, advantages that the GnRH receptor has high expressionin a reproductive system tumor such as ovarian cancer, the near-infrared fluorescent dye ICG penetrates deeper and the background tissue autofluorescence is weaker are utilized, tumor cells and tumor foci which cause high expressionof the GnRH receptor can be targeted and recognized. The GnRHreceptor-targeted near-infrared fluorescent imaging probe GnRHa-ICG has a good application prospect in fluorescence imaging and fluorescence guided surgery.

Description

Technical field [0001] The invention relates to the field of biotechnology, in particular to a preparation method and application of a tumor-specific fluorescent probe for treating malignant tumors of the reproductive system. Background technique [0002] Malignant tumors are an important threat to human health. It is estimated that there will be more than 18 million new cancer cases and 9.6 million cancer deaths worldwide in 2018. Among reproductive system-related malignancies, prostate cancer accounted for 13.5% of new tumor cases in men, and breast cancer, uterine body tumors, and ovarian cancer accounted for 24.2%, 4.4%, and 3.4% of new cases in women. [0003] Surgery is an important treatment for solid tumors, and the thoroughness of surgery is closely related to the prognosis. In order to achieve a complete resection, accurate identification of the lesion during the operation is the key. The existing imaging methods such as ultrasound, CT, MRI, etc. are non-specific imagi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00C07K16/28C07K1/06C07K1/04
CPCA61K49/0034A61K49/0056C07K16/2869
Inventor 张晓燕刘齐雨周晓波李小平李富友徐丛剑
Owner THE OBSTETRICS & GYNECOLOGY HOSPITAL OF FUDAN UNIV
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