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Myogenesis inducing agent

A differentiation inducer and muscle technology, which can be applied in the fields of muscle system diseases, neuromuscular system diseases, organic active ingredients, etc., can solve the problems of decreased muscle differentiation ability and difficult treatment methods, and achieve the effect of restoring muscle strength and enhancing muscle strength.

Inactive Publication Date: 2019-07-23
NAT CENT OF NEUROLOGY & PSYCHIATRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, a therapeutic approach with exercise is difficult to implement in the bedridden elderly and in patients with myogenic diseases with severe muscle wasting
In addition, aging in the elderly leads to a decline in muscle differentiation, so exercise therapy alone cannot be expected to suppress muscle atrophy and increase skeletal muscle mass

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0158]

[0159] (Purpose)

[0160] The muscle differentiation-inducing activity of miR-199a-3p constituting the muscle differentiation-inducing agent of the present invention on myoblasts was verified.

[0161] (method and results)

[0162] After introducing miRNAs of miR-199a-3p, miR-1, miR-133a, miR-26a, miR-214, miR-125a, miR-133b and Let-7e into mouse myoblast cell line C2C12 cells by electroporation , the cells were inoculated into muscle differentiation medium (2% HS / DMEM) and cultured at 30°C for 3 days, and then the total RNA of the cells was used to verify the muscle differentiation marker proteins myogenin (Myog) and The expression level of Myh1.

[0163] The results are shown in figure 1 . A is the expression level of Myog, and B is the expression level of Myh1. Among these two markers, the expression of muscle differentiation markers in C2C12 cells introduced with miR-199a-3p was significantly higher than that of C2C12 cells introduced with other miRNAs. Th...

Embodiment 2

[0164]

[0165] (Purpose)

[0166] The expression levels of muscle differentiation-related miRNAs in C2C12 cells after introduction of miR-199a-3p constituting the muscle differentiation inducer of the present invention were verified.

[0167] (method and results)

[0168] U6snRNA was used as an internal standard gene and analyzed by ΔΔCt method to determine the expression levels of miR-1, miR-133a, miR-26a, let-7g and miR-214 in C2C12 cells after miR-199a-3p was introduced. The imported miR-199a-3p used MISSION miRNA mimic miR-199a-3p (HMI0338) (referred to as "199s") and miR-199a-3p#4 (referred to as "199#4").

[0169] The results are shown in Figure 2. Such as diagram 2-1 As shown, the expression levels of miR-1 in 199s and 199#4 were significantly higher than those in the control (neg). In addition, if Figure 2-2 As shown, the expression of miR-1 and miR-133a in C2C12 cells introduced with 199s was significantly increased compared with the control (neg). This res...

Embodiment 3

[0170]

[0171] (Purpose)

[0172] According to the base sequence of miR-199a-3p, it can be deduced that the target candidate genes of miR-199a-3p are Lin28b gene and Suz12 gene. Such as Figure 3-1 As shown, mouse miR-199a-3p has two candidate target sites on the Lin28b gene (A: 81st to 102nd, and B: 983rd to 1004th); in addition, on the Suz12 gene There is also a candidate target site (C: 973rd to 994th). Therefore, it was verified whether the expression of the Lin28b gene and the Suz12 gene, which are target candidate genes, were suppressed in the cells when miR-199a-3p was introduced into the C2C12 cells.

[0173] (method and results)

[0174] After introducing miR-199a-3p into C2C12 cells and culturing them, cell extracts were prepared according to conventional methods. Western blot was performed on the cell extract, and the expression levels of Lin28b protein and Suz12 protein were detected by using antibodies against Lin28b protein and Suz12 protein respectively. ...

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Abstract

Provided is an agent developed to be capable of alleviating and suppressing muscle atrophy or muscle mass reduction, even for the elderly and even without requiring exercise, by inducing myogenesis. Provided is a composition for treating or preventing disorders or diseases associated with muscle atrophy, or for promoting muscle regeneration, the composition containing, as an active ingredient, a myogenesis inducing agent composed of miR-199 or an DNA that contains an miR-199 gene encoding the miR-199.

Description

technical field [0001] The present invention relates to an agent for inducing muscle differentiation, and a composition for treating or preventing disorders or diseases accompanied by muscle atrophy or muscle damage or a composition for promoting muscle regeneration comprising the agent for inducing muscle differentiation as an active ingredient. Background technique [0002] In recent years, many major developed countries have rapidly entered the era of super-aging societies, and the average life expectancy of both men and women exceeds 80 years. For example, the average life expectancy in Japan in 2015 was 80.50 years for men and 86.83 years for women. It is predicted that the increase of the elderly population will continue for more than 20 years in the world in the future. [0003] As many countries usher in a super-aging society, various problems associated with aging have also surfaced. For example, social problems such as age-related restrictions on daily life activ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7105A61P21/00C12N15/113
CPCA61K31/7105A61P21/00C12N15/113C12N2310/141
Inventor 北条浩彦福冈圣之
Owner NAT CENT OF NEUROLOGY & PSYCHIATRY
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