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Novel application of PTP1B (protein tyrosine phosphates 1b) inhibitor

A technology of inhibitors and uses, applied in the field of male infertility, can solve problems such as the inability of leptin to play

Inactive Publication Date: 2014-06-11
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But leptin doesn't have the same effect in the human body

Method used

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  • Novel application of PTP1B (protein tyrosine phosphates 1b) inhibitor
  • Novel application of PTP1B (protein tyrosine phosphates 1b) inhibitor
  • Novel application of PTP1B (protein tyrosine phosphates 1b) inhibitor

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Experimental program
Comparison scheme
Effect test

Embodiment 1～8

[0094] general method

[0095] 1. Detection of PTP1B expression in sperm

[0096] (1) Sperm with higher motility were collected by the upstream method, and capacitated in the incubator for 1 hour.

[0097](2) Centrifuge at room temperature for 10 minutes at a speed of 1,500 g.

[0098] (3) Protein lysate was lysed, centrifuged at 4°C for 5min at a rotational speed of 12,000g, the supernatant was taken, and loaded with loading buffer at 100°C for 5min.

[0099] (4) The expression level of PTP1B was detected by immunoblotting.

[0100] (5) Perform data analysis.

[0101] 2. Detection of PTP1B activity in sperm

[0102] (1) Obtain protein lysate: lyse the sperm at 4°C at a rotational speed of 12,000g, centrifuge at 4°C for 5 minutes to take the supernatant, and take 200 μg of total protein from each tube.

[0103] (2) Pre-purified protein solution:

[0104] a. Add 50μl normal rabbit serum to every 1ml protein solution;

[0105] b. Incubate on ice for 1 hour;

[0106] c. E...

Embodiment 1

[0129] Differential detection of PTP1B expression in animal models

[0130] Use Western Blot technology to detect the difference in the expression of PTP1B protein in the sperm of 12-week-old normal mice (C57BL / 6J, purchased from and obese mice. The method is as follows:

[0131] The two groups of mice were sacrificed by neck dislocation and the epididymis were taken out. The epididymis was cut open and the sperm were extruded into conventional HTF medium (purchased from Irvine Scientific).

[0132] The results showed that the expression of PTP1B in the sperm of 12-week-old obese mice was 3-4 times higher than that of normal mice ( figure 1 ).

Embodiment 2

[0134] Inhibition of PTP1B by PTP1B inhibitors

[0135] In this embodiment, the activity of PTP1B in mouse sperm before and after administration was tested, the method is as follows:

[0136] (1) Mouse sperm capacitation

[0137] Under the condition of 37°C, normal sperm and obese mouse sperm were capacitated in conventional HTF medium for 2 hours.

[0138] (2) PTP1B inhibition

[0139] Ca 2+ It flows into the sperm membrane, thereby activating the downstream Rab3A pathway for acrosome reaction. In this experiment, each group was incubated with various concentrations of PTP1B inhibitors for 10 minutes, and then added 15 μM progesterone for 15 minutes to stimulate the acrosome reaction of mouse sperm.

[0140] The activity of PTP1B protein in the sperm of normal mice and obese mice was detected by a general method.

[0141] See the experimental results figure 2 . The results showed that PTP1B activity was positively correlated with protein expression, and after using PT...

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Abstract

The invention discloses application of a PTP1B (protein tyrosine phosphates 1b) inhibitor and a PTP1B gene, and in particular relates to the PTP1B inhibitor in preparation of a medicine for treating infertility (a), and a medicine for improving acrosome reaction (b) and or improving fertility rate (c). Experiments indicate that: the PTP1B inhibitor can be used for effectively improving the acrosome reaction and fertility rate. According to the application, infertility patients with obesity or normal body weight can be screened by detecting whether a PTP1B is highly expressed in a body or not, so that a treatment scheme for the infertility patients can be determined, and is more flexible, easier and more economical that that of common diagnostic treatment in a traditional infertility treatment process.

Description

technical field [0001] The invention relates to the application of a PTP1B inhibitor in the preparation of medicaments for treating male infertility, especially male infertility in obese patients and high-expression groups of PTP1B gene. Background technique [0002] At present, there are more and more cases of infertility caused by various reasons, among which male infertility is one of the important causes of infertility. [0003] Studies have shown that in infertile men, there are still some expression changes in the body, including proteins whose expression is up-regulated or down-regulated. Among them, some up-regulated proteins include: retinitis pigmentosa GTPase regulator (RPGR) hinders flagella assembly; follistatin (follistatin), human liver regeneration enhancer (hALR), although these highly expressed proteins are associated with male infertility symptoms There seems to be some correlation, however, so far, there have been no reports of successful treatment of ma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61P15/08C12N5/076C12Q1/68
Inventor 曾科石磊张辰宇张骑鹏
Owner NANJING UNIV
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