A near-infrared fluorescent imaging probe targeting gnrh receptors and its preparation method and application

Abstract

The invention provides a GnRH receptor-targeted near-infrared fluorescence imaging probe GnRHa-ICG, a preparation method and application thereof. The near-infrared fluorescent imaging probe GnRHa-ICG targeting the GnRH receptor consists of a GnRH polypeptide (sequence D-2-Nal-D-4-Cl-Phe-D-3-Pal-Ser-Tyr-D-Cit- Leu‑Arg‑Pro‑D‑Ala‑NH 2 ) and activated near-infrared fluorescent dye ICG-NHS coupling. The present invention is based on the principle of specific binding between GnRH polypeptides and GnRH receptors, and utilizes the high expression of GnRH receptors in reproductive system tumors such as ovarian cancer, and the advantages of deeper penetration depth of near-infrared fluorescent dye ICG and weaker background tissue autofluorescence. Targeted recognition of tumor cells and tumor foci that highly express GnRH receptors 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. Worldwide, more than 18 million new cancer cases and 9.6 million cancer-related deaths are expected in 2018. Among reproductive system-related malignancies, prostate cancer accounted for 13.5% of new cases in men, and breast, uterine and ovarian cancers accounted for 24.2%, 4.4%, and 3.4% of new cases in women, respectively.

[0003] Surgery is an important treatment for solid tumors, and the thoroughness of surgery is closely related to prognosis. In order to achieve complete resection, accurate identification of the lesion during surgery is the key. Existing imaging methods such as ultrasound, CT, and MRI are non-specific imaging of tumors, and it is difficult t...

Examples

Embodiment

[0036] The specific embodiments of the application of the above-mentioned near-infrared fluorescent probes in tumor targeting imaging are as follows:

[0037] The human ovarian cancer cell line A2780 (high expression of GnRH receptor) and human lung cancer cell line H1299 (low expression of GnRH receptor) in logarithmic growth phase were inoculated on chamber slides, and the experiment was started when the cell confluence reached 50%. . The culture medium was aspirated and washed once with PBS buffer. 60 μM of GnRHa-ICG and ICG were added, respectively, and incubated with tumor cells for 30 min at 37°C. Aspirate the supernatant and wash 3 times with PBS. 4% paraformaldehyde was added for fixation at room temperature for 10 min. Aspirate the fixative and wash once with PBS. The cell membrane dye WGA488 was added and incubated at room temperature for 10 min. The membrane dye was aspirated and washed 3 times with PBS. Mount the slides with DAPI-containing fluorescent mounti...

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. Worldwide, more than 18 million new cancer cases and 9.6 million cancer-related deaths are expected in 2018. Among reproductive system-related malignancies, prostate cancer accounted for 13.5% of new cases in men, and breast, uterine and ovarian cancers accounted for 24.2%, 4.4%, and 3.4% of new cases in women, respectively.

[0003] Surgery is an important treatment for solid tumors, and the thoroughness of surgery is closely related to prognosis. In order to achieve complete resection, accurate identification of the lesion during surgery is the key. Existing imaging methods such as ultrasound, CT, and MRI are non-specific imaging of tumors, and it is difficult t...

Examples

Embodiment

[0036] The specific embodiments of the application of the above-mentioned near-infrared fluorescent probes in tumor targeting imaging are as follows:

[0037] The human ovarian cancer cell line A2780 (high expression of GnRH receptor) and human lung cancer cell line H1299 (low expression of GnRH receptor) in logarithmic growth phase were inoculated on chamber slides, and the experiment was started when the cell confluence reached 50%. . The culture medium was aspirated and washed once with PBS buffer. 60 μM of GnRHa-ICG and ICG were added, respectively, and incubated with tumor cells for 30 min at 37°C. Aspirate the supernatant and wash 3 times with PBS. 4% paraformaldehyde was added for fixation at room temperature for 10 min. Aspirate the fixative and wash once with PBS. The cell membrane dye WGA488 was added and incubated at room temperature for 10 min. The membrane dye was aspirated and washed 3 times with PBS. Mount the slides with DAPI-containing fluorescent mounti...

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