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Application of ctbs protein as the target of sperm-egg binding receptors in the screening of sperm-egg binding inhibitors

A technology of sperm-egg binding receptors and inhibitors, applied in the field of application of CTBS protein as the target of sperm-egg binding receptors in the screening of sperm-egg binding inhibitors

Inactive Publication Date: 2015-11-18
SHANGHAI INST OF PLANNED PARENTHOOD RES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been reported that the oligosaccharide LewisX can bind to mouse sperm and specifically inhibit the in vitro fertilization of mice, suggesting that the protein that recognizes LewisX on the sperm may be involved in the reproductive isolation of mice [9] , but only a guess

Method used

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  • Application of ctbs protein as the target of sperm-egg binding receptors in the screening of sperm-egg binding inhibitors
  • Application of ctbs protein as the target of sperm-egg binding receptors in the screening of sperm-egg binding inhibitors
  • Application of ctbs protein as the target of sperm-egg binding receptors in the screening of sperm-egg binding inhibitors

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] We found that CTBS is expressed in rodent sperm, and CTBS antibody can inhibit in vitro fertilization in mice, but this protein is not expressed in human sperm. Its experimental method is as follows:

[0042] (1) Discovery of CTBS on mouse spermatozoa:

[0043] The CTBS on the mouse sperm was identified by using the Lewis oligosaccharide X (ie LewisX, abbreviated as LeX) probe, and it was proved that CTBS can bind to the zona pellucida of the mouse egg.

[0044] (1) First identify the LewisX receptor, the identification process is as follows Figure 2A As shown, it includes the identification process of lectin-like proteins that can recognize LewisX and the identification process of endogenous LewisX-modified proteins. The principle is to modify the surface of bovine serum albumin (BSA) with LewisX to make the pseudoglycoprotein LeX-BSA, and further modify the pseudoglycoprotein with ultraviolet crosslinking reagent [6-(biotinamido)-2-(p -Azidobenzamido)caproylamino]...

Embodiment 2

[0067] Embodiment 2: Dequalinium chloride is used as the inhibitor experiment of suppressing mouse sperm-egg combination

[0068] Dequalinium chloride was used as an inhibitor to inhibit the combination of sperm and eggs in mice, and each dose of CTBS with different concentrations was used to experiment with more than 50 mouse eggs, repeated 3 times, and each dose used a total of more than 150 eggs. The experimental results Such as figure 1 As shown, its half-inhibition constant is at 50nM, indicating that dequalinium chloride can well inhibit the in vitro fertilization of mice.

[0069] In summary, all the results described in Example 1 and Example 2 show that CTBS is a receptor protein specifically expressed on rodent sperm, which participates in the physiological process of sperm-egg binding, and the CTBS antibody can inhibit the mouse in vitro Fertilize. As a birth control drug for mice, dequalinium chloride can well inhibit the in vitro fertilization of mice, and experi...

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Abstract

The invention relates to an application of a CTBS (Di-N-acetylchitobiase) protein serving as a target spot of a sperm-egg binding acceptor to screening of a sperm-egg binding inhibitor. According to the application, CTBS is a special sperm-egg binding acceptor on rodent sperm; the CTBS protein only has the specific expression on the sperms of rodents and the protein is not expressed on sperms of human beings; a CTBS antibody can inhibit the sperm-egg binding of mice. The screened sperm-egg binding acceptor is dequalinium chloride and the semi-inhibition constant is less than 100nM. The dequalinium chloride serves as fertility regulation medicine of the mice and the sperm-egg binding of the rodents can be effectively controlled, thus controlling the population quantity of the mice; the aim of controlling mouse damages without pesticides is realized; the damages are not caused when natural enemies of the mice eat the medicine.

Description

technical field [0001] The invention relates to the application of a CTBS protein as a target of sperm-egg binding receptors in screening sperm-egg binding inhibitors. Background technique [0002] Dequalinium chloride (Dequaliniumchloride, CAS#522-51-0), used as a PKC inhibitor as an anticancer drug, its IC50 is 7-18μM, and it is also a selective alittin-sensitive potassium Ion channel blocker with an IC50 of 1.1 μM [1,2，3,4] . In recent years, dequalinium chloride has been used as a cationic surfactant in non-antibiotic antibacterial drugs (the drug name is "clotidine"). Recently, it was found that dequalinium chloride can also inhibit the activity of microbial-derived chitin hydrolase at the nanomolar level [5] . Dequalinium chloride has little toxicity to the human body, does not affect human reproduction, and is irritating to the eyes and skin. The lethal dose to mice is: oral administration of 150-1000 mg / kg body weight, subcutaneous injection of 70 mg / kg body wei...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
CPCG01N33/68G01N33/94
Inventor 顾一骅徐文平施惠娟杨军
Owner SHANGHAI INST OF PLANNED PARENTHOOD RES
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