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Judgment or evaluation method of test substance

An evaluation method and extraction technology, applied in biochemical equipment and methods, microbial determination/testing, biological testing, etc.

Active Publication Date: 2015-09-30
NIPPON ZOKI PHARM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, for the first time, the promotion of BDNF expression and its regulatory effect of the inflamed tissue extract inoculated with vaccinia virus were clarified, and the promotion of BDNF expression and the activation of various proteins in the intracellular signal transduction pathway related to it were used as indicators to evaluate the expression of BDNF. The method of judging or evaluating the extract is unknown so far

Method used

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  • Judgment or evaluation method of test substance
  • Judgment or evaluation method of test substance
  • Judgment or evaluation method of test substance

Examples

Experimental program
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Effect test

Embodiment 1

[0066] Inoculate vaccinia virus on the skin of healthy mature rabbits, peel off the acne-prone skin, break it up and add phenol water. Then, this was filtered under pressure, and the obtained filtrate was adjusted to pH 5 with hydrochloric acid, and then heat-treated at 90-100° C. for 30 minutes. After removing the protein by filtration, adjust the pH to 9 with sodium hydroxide, heat treatment at 90-100° C. for 15 minutes, and then filter. The filtrate was adjusted to about pH 4.5 with hydrochloric acid, 2% activated carbon was added, stirred for 2 hours and then centrifuged. Add water to the collected activated carbon, adjust the pH to 10 with sodium hydroxide, stir at 60° C. for 1.5 hours, centrifuge and filter to obtain the supernatant. Add water to the collected activated carbon again, adjust the pH to 11 with sodium hydroxide, stir at 60° C. for 1.5 hours, and centrifuge to obtain the supernatant. The two supernatants were combined, neutralized with hydrochloric acid, a...

Embodiment 2

[0068] After inoculating the skin of healthy mature rabbits with vaccinia virus to infect them, the acne-prone skin was aseptically peeled off, minced, added to glycerin water with phenol, and ground into milky form with a homogenizer. Then, this was filtered, and the obtained filtrate was adjusted to weakly acidic (pH 4.5 to 5.5) with hydrochloric acid, then heated at 100° C., and filtered. The filtrate was adjusted to be weakly alkaline (pH 8.5 to 10.0) with sodium hydroxide, then heated at 100°C and filtered. The filtrate was adjusted to about pH 4.5 with hydrochloric acid, about 1.5% activated carbon was added, stirred for 1 to 5 hours, and then filtered. Add water to the filtered activated carbon, adjust the pH to 9.4 to 10 with sodium hydroxide, filter after stirring for 3 to 5 hours, and neutralize the filtrate with hydrochloric acid.

Embodiment 3

[0070] Inoculate the skin of healthy mature rabbits with vaccinia virus, activate it, peel off the activated skin aseptically, chop it up, add water, grind it into milky form with a homogenizer. Then, this was filtered under pressure, and the obtained filtrate was adjusted to pH 5.0 with hydrochloric acid, and then heat-treated at 100° C. under steam circulation. After the protein was removed by filtration, the pH was adjusted to 9.1 with sodium hydroxide, followed by heat treatment at 100°C and filtration. The filtrate was adjusted to pH 4.1 with hydrochloric acid, 2% activated carbon was added, stirred for 2 hours and then filtered. Add 5.5% activated carbon to the filtrate again, filter after stirring for 2 hours. Water was added to the activated carbon collected by filtration, the pH was adjusted to 9.9 with sodium hydroxide, and the mixture was stirred at 60° C. for 1.5 hours and then filtered. Water was added to the first and subsequent activated carbons, adjusted to p...

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Abstract

An object of the present invention is to provide a method for determination or evaluation of an extract from inflamed tissues inoculated with vaccinia virus where the enhancement of activation of neurotrophic factor such as BDNF in cultured cells or the enhancement of activation of proteins participating therein is used as an indicator. The present invention relates to a novel method for determination or evaluation of an extract from inflamed tissues inoculated with vaccinia virus and relates to a method for determination or evaluation of the extract where the enhancement of production of neurotrophic factor such as BDNF in cultured cells or the enhancement of activation of various proteins in intracellular signaling pathway participating in production of BDNF, etc. is used as an indicator. The present invention is highly useful as a simple, convenient and quick method for determination or evaluation for guaranteeing the quality of the extract which is useful as a pharmaceutical agent.

Description

technical field [0001] The present invention relates to a novel determination or evaluation method of an extract of an inflammatory tissue inoculated with vaccinia virus, and more specifically, to the effect of an extract of an inflammatory tissue inoculated with vaccinia virus as a test substance on the expression of BDNF in cultured cells or on the expression of BDNF in cultured cells. A method of judging or evaluating the test substance by using the actions of various proteins in the intracellular signal transduction pathway related to BDNF production as indicators. In addition, "treating" herein means culturing cells in the presence of the substance (hereinafter also the same). Background technique [0002] Brain-derived neurotrophic factor (BDNF) and nerve growth factor (nerve growth factor: NGF) and neurotrophin-3 (neurotrophin-3: NT-3) belong to the neurotrophin family (neurotrophic factor family ), is a secreted protein produced by various cells such as nerve cells ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68C12Q1/00
CPCG01N33/5023C12Q1/6876G01N33/5041G01N2333/07G01N33/6854
Inventor 武藤多津郎福田有
Owner NIPPON ZOKI PHARM CO LTD
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