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Drug for nerve regeneration

A technology for nerve regeneration and neurological diseases, which can be used in neurological diseases, drug combinations, pharmaceutical formulations, etc., and can solve the problems of treating neurodegenerative diseases and ignorance

Inactive Publication Date: 2006-05-17
KYOWA HAKKO KOGYO CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to this report, it is considered that substances that can inhibit the activity of GSK-3 can be used as therapeutic drugs for various neurodegenerative diseases represented by Alzheimer's disease (International Patent Application Publication No. 00 / 38675 ), however, it is not known whether neurodegenerative diseases can actually be treated by protecting mature neurons, and whether substances that inhibit GSK-3 activity have the effect of promoting neuronal regeneration

Method used

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  • Drug for nerve regeneration
  • Drug for nerve regeneration
  • Drug for nerve regeneration

Examples

Experimental program
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Effect test

experiment example 1

[0167] Experimental example 1: Lithium chloride promotes neuron regeneration (1)

[0168]According to the method of the above-mentioned 7, when ANSC-7 cells are differentiated and induced, lithium chloride or sodium chloride (both are manufactured by Nacalai Tesque Co., Ltd.) The solution was added to the culture medium containing ANSC-7 cells according to the volume of 1 / 1000 of the culture medium, and the number of neurons therein was analyzed on the 6th day after differentiation induction. As a result, the number of Tuj1-positive neurons was 1.1, 1.3, 1.8, and 2.1 times in the solutions with final concentrations of lithium chloride of 0.01, 0.1, 1, and 3 mmol / l, respectively (when the concentration of lithium chloride was above 1 mmol / l There is a significant difference), that is, the number of neurons increases with the increase of lithium chloride concentration. Moreover, compared with the control sample without lithium chloride, the total number of H33342-positive cells...

experiment example 2

[0169] Experimental example 2: Promotion of regeneration of neurons by lithium chloride (2)

[0170] In order to find out whether the promoting effect on neuron regeneration of ANSC-7 cells is caused by the expression induction of BDNF and Bcl-2, semi-quantitative RT-PCR method was used to analyze whether lithium can promote the expression of BDNF and Bcl-2.

[0171] Using a 6-well culture dish coated with polyornithine and laminin, add 2ml of DMEM / F12 medium containing 1% N2 supplement and 20ng / ml FGF-2 to it, and then add to the culture dish Inoculate 4.5 x 10 within each well 5 A total of 7 wells were inoculated with ANSC-7 cells, and cultured overnight. Then, total RNA was obtained from one well of cells using RNeasy mini kit (manufactured by QIAGEN) according to the attached instructions. The medium in the remaining 6 wells was all replaced with differentiation induction medium to induce differentiation. 3 mol / l lithium chloride equivalent to 1 / 1000 volume of the mediu...

experiment example 3

[0178] Experimental example 3: Lithium chloride promotes neuron regeneration (3)

[0179] In order to ascertain whether the neuronal regeneration promotion effect induced by lithium is the result of the increase in the number of regenerated cells due to the inhibition of apoptosis, or the result of actively inducing neuronal differentiation, the inhibitory effect of ANSC-7 induced by lithium The effect of apoptosis was analyzed.

[0180] According to the method of 7 above, lithium chloride was added to the medium containing ANSC-7 cells to a final concentration of 3 mmol / l, and they were cultured for 6 days, using the in situ cell death detection kit and fluorescent yellow (Roche Diagnostics K , manufactured by K.), and reacted with ANSC-7 cells cultured in the addition of lithium chloride and ANSC-7 cells cultured in the addition of PBS as a control according to the accompanying instructions. Use an inverted fluorescence microscope (manufactured by Nikon Corporation) to obse...

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Abstract

An object of the present invention is to provide a nerve regeneration drug, a neuron regeneration promoter of neural stem cells, a neuron obtained by culturing neural stem cells in the presence of the neuron regeneration promoter, and a method for producing the neuron. In order to achieve this object, the present invention provides a nerve regeneration drug containing a substance that inhibits the activity of glycogen synthase kinase-3 as an active ingredient, a neural stem cell neuron regeneration promoter containing the substance as an active ingredient, and A neuron obtained by culturing neural stem cells in the presence of a neuron regeneration promoting agent and a method for producing the neuron. Medicine of the present invention can be used as Parkinson's disease, Alzheimer's disease, Down's syndrome, cerebrovascular disorder, cerebral apoplexy, spinal cord injury, Huntington's chorea, multiple sclerosis, amyotrophic lateral sclerosis It is used in the treatment of neurological diseases such as neurological disorders, epilepsy, anxiety disorders, complex disorders, depression, and bipolar disorder.

Description

[0001] Technical division [0002] The present invention relates to a nerve regenerative drug containing a substance inhibiting the activity of glycogen synthase kinase-3 (hereinafter abbreviated as GSK-3) as an active ingredient, and a neuron regeneration promoter containing a substance inhibiting the activity of GSK-3 as an active ingredient , a neuron obtained by culturing neural stem cells in the presence of the neuron regeneration promoting agent, and a method for producing the neuron. Background technique [0003] Neurological disease is a general term for neurodegenerative diseases, depression and bipolar disorder without neurodegeneration, and these diseases are diseases caused by damage to the brain or peripheral neurons due to genetic factors, environmental factors, and aging factors. Specific examples of neurodegenerative diseases include Parkinson's disease, Alzheimer's disease, polyglutamic acid disease, amyotrophic lateral sclerosis, polyneuropathy, spinal cord i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K31/00A61K31/404A61K31/4045A61K31/407A61K31/553A61P25/08A61P25/18A61P25/22A61P25/24A61P25/28C07D403/04C07D403/14C07D498/22C12N5/07C12N5/079C12N5/0797
CPCA61K31/00A61K31/404A61K31/4045A61K31/407A61K31/553A61P25/00A61P25/08A61P25/14A61P25/16A61P25/18A61P25/22A61P25/24A61P25/28A61P43/00A61P9/00
Inventor 森下刚樱田一洋铃木惠子池田俊一
Owner KYOWA HAKKO KOGYO CO LTD
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