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Follicular granular cells near-infrared fluorescent probe Nirova-2 as well as preparation method and application thereof

A technology of granule cells and fluorescent probes, applied in the direction of pharmaceutical formulations, preparations for in vivo tests, etc., to achieve the effects of good stability and repeatability, simple preparation methods, and high penetration depth

Inactive Publication Date: 2020-04-24
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, it has not been reported to use IR800 CW and the probe synthesized by human follicle-stimulating hormone to fluorescently label follicular granulosa cells in the near-infrared region of the ovary

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Weigh IR800 CW Maleimid (1.5mg, 1.3µmol) and dissolve it in DMSO, and dissolve 25 µg of human follicle-stimulating hormone (FSH) in 0.1mL1X phosphate buffer (PBS). After adding 1mM dithiothreitol (DTT) to the FSH-PBS system for pretreatment, add the above solution system into a 1 mL glass reactor so that the molar ratio of dye to protein is 100:1, and stir at 4°C for 20h , turn such as 3kDa (Amicon) ultrafiltration tube, 4500 rpm, 15 minutes centrifugal dialysis, repeat this step twice with 1X phosphate buffer solution.

Embodiment 2

[0025] Weigh IR800 CW Maleimid (1.5mg, 1.3µmol) and dissolve it in DMSO, and dissolve 25 µg of human follicle-stimulating hormone (FSH) in 0.1mL1X phosphate buffer (PBS). After the FSH-PBS system was pretreated by adding 1 mM tris(2-carboxyethyl)phosphine (TCEP), the above solution system was added to a 1 mL glass reactor so that the molar ratio of dye to protein was 100:1. Stir at constant temperature for 20 hours at ℃, turn to a 3kDa (Amicon) ultrafiltration tube, 4500 rpm, and centrifugally dialyze for 15 minutes. Repeat this step twice with 1X phosphate buffer solution.

Embodiment 3

[0027] Weigh IR800 CW Carboxylate (1.4mg, 1.3µmol) and dissolve it in DMSO, and dissolve 25 µg of human follicle-stimulating hormone (FSH) in 0.1mL1X phosphate buffer (PBS). Add the above solution system into a 1 mL glass reactor, and add 2.3 µLEDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) stock solution at the same time to make the dye and protein in the system The molar ratio of the dye to EDC is 100:1, the molar ratio of the dye to EDC is 1:1, stirred at 4°C for 20 hours, transferred to a 3kDa (Amicon) ultrafiltration tube, 4500 rpm, 15 minutes centrifugal dialysis, and 1X phosphoric acid Repeat this step twice for saline buffer solution.

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PUM

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Abstract

The invention belongs to the technical field of medical imaging contrast agents, and particularly relates to a follicular granular cells near-infrared fluorescent probe Nirova-2 and a preparation method and application thereof. The fluorescent probe provided by the invention is a fluorescent dye-human follicle stimulating hormone coupling complex formed by coupling an organic small molecule near-infrared dye IR800CW and human follicle stimulating hormone (FSH); according to the complex, the fluorescent dye can specifically target granular cells forming follicle walls in ovaries, so that near-infrared fluorescent labeling of all follicles is realized. The fluorescent probe can be used as a near-infrared region fluorescent contrast agent for the follicles in the ovaries by combining the characteristic of fluorescence emission of the IR800CW in a near-infrared region with the property of specific targeted binding of the FSH to the follicle granular cells in the ovaries, so that the numberand the development stage of the follicles in the ovaries of women can be accurately and effectively detected and evaluated.

Description

technical field [0001] The invention belongs to the technical field of medical imaging contrast agents, and in particular relates to a near-infrared fluorescent probe for follicular granulosa cells and a preparation method and application thereof. Background technique [0002] At present, it is often necessary to detect mature and immature follicles in the ovaries for women of childbearing age or overage who suffer from infertility. The current implementation methods are traditional B-mode ultrasound imaging technology or transvaginal ultrasound imaging technology. However, this technique can only observe follicles above the second grade (usually above 600 μm), limited by the wavelength of sound waves, the highest resolution of ultrasound imaging is at the level of 100 microns, so for smaller follicles (such as primordial follicles, the diameter Only a few microns) cannot be detected, resulting in some patients with only small residual follicles losing hope for further treat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00
CPCA61K49/0056A61K49/0021
Inventor 张凡冯异朱昕燕
Owner FUDAN UNIV
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