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Application of methyl 3, 4-dihydroxybenzoate in preparation of medicaments for preventing and treating nerve degenerative diseases

A technology of methyl dihydroxybenzoate and medicine, which is applied in the application field of methyl 3,4-dihydroxybenzoate in the preparation of medicines for preventing and treating neurodegenerative diseases, can solve the problems of unseen neuron growth and the like, and achieve prevention Loss of synapses, maintenance of neuronal survival, effects of stable culture conditions

Active Publication Date: 2014-05-14
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, it has been reported in the literature that it has antioxidant activity, but there is no report on its promotion of neuron growth

Method used

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  • Application of methyl 3, 4-dihydroxybenzoate in preparation of medicaments for preventing and treating nerve degenerative diseases
  • Application of methyl 3, 4-dihydroxybenzoate in preparation of medicaments for preventing and treating nerve degenerative diseases
  • Application of methyl 3, 4-dihydroxybenzoate in preparation of medicaments for preventing and treating nerve degenerative diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 3,4-Dihydroxybenzoic acid methyl ester promotes the growth of rat cortical neurons

[0031] 1. Primary Culture of Rat Cortical Neurons

[0032] Take the required number of newborn mice and soak them in 75% ethanol for 20 seconds. Cut the skin and skull, place the removed brain in D-Hanks buffer, separate the meninges and hippocampus, and keep the cortex. Transfer the cortex into a vial of penicillin containing about 2ml of D-Hanks buffer, cut it into pieces with ophthalmic scissors, add 1mL of 0.25% protease to the vial, and put it in an incubator for digestion for 10 minutes. Digestion was terminated with DMEM / F12 containing 10% FBS. Filter with a 400-mesh filter, collect the filtrate, transfer it to a 15ml centrifuge tube, and centrifuge at 1000 r / min for 5 minutes. Remove the supernatant, add 2 mL inoculum solution (DMEM / F12 containing 0.4% B-27), and pipette gently for about 30 times to make it into a cell suspension. Take 50 μL of cell suspension ...

Embodiment 2

[0041] Example 2 Effect of methyl 3,4-dihydroxybenzoate on the activity of rat cortical neurons

[0042] Divide neurons into 1.2×10 5 cells / cm 2 The density was seeded in a 96-well plate, and when the medium was changed 5 hours later, it was divided into solvent control group (DMSO), positive drug control group (BDNF) and test drug (MDB) dose groups (2μM, 4μM, 8μM and 16μM), Each group has 6 replicate wells. After continuing to culture for 72 hours, remove the medium, add 100 μL DMEM / F12+10 μL MTT to each well, and continue to culture for 4 hours. Aspirate the medium, add 150 μL DMSO, shake for 10 minutes, and detect on a microplate reader at a wavelength of 570 nm. Results (see Figure 4 ) showed that the positive control drug (BDNF) and different concentrations of 3,4-dihydroxybenzoic acid methyl ester were added to the culture medium and cultured for 72 hours. Compared with the solvent control group, BDNF (20ng / ml ) group and the test drug (2μM ~ 16μM) can increase ...

Embodiment 3

[0043] Example 3 Effect of methyl 3,4-dihydroxybenzoate on the expression of MAP2 mRNA in rat cortical neurons

[0044] (1) Design and synthesis of primers

[0045] According to the sequence of Genebank, primers for the target gene and internal reference gene were designed with Primer 5.0 software, and were synthesized by Yingwei Jieji (Shanghai) Trading Co., Ltd. (Table 1).

[0046] (2) Extraction of total cellular RNA

[0047] Divide neurons into 1.2×10 5 cells / cm 2 Inoculated in a 6-well plate at a density of 5 hours, and divided into solvent control group (DMSO), positive drug control group (BDNF 20ng / ml) and test drug (MDB) dose groups (2μM, 4μM, 8μM) when the medium was changed 5 hours later. . One well for each group, and the experiment was repeated three times. Neurons were cultured until the third day, and the cultured cells of each group were collected, lysed by adding 1 mL Trizol RNA extraction reagent, extracted with 200 μL chloroform, centrifuged at high s...

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Abstract

The invention discloses the application of methyl 3, 4-dihydroxybenzoate (MDB) in preparation of medicaments for preventing and treating nerve degenerative diseases. According to the invention, MDB or derivatives thereof have double functions of maintaining survival of neurons or nerve cells and promoting growth of nervous processes, can be applied to develop neurotrophic factor mimetic agents for treating nerve degenerative diseases (such as AD and PD) and other related diseases, and can also be applied to cell and gene transplantation therapy for nerve degenerative diseases (such as AD and PD) and other related diseases. The invention determines new pharmacological activity of MDB and illuminates pharmacodynamic mechanism of MDB, and provides reference for developing new drugs with independent intellectual property rights.

Description

technical field [0001] The invention relates to the application of methyl 3,4-dihydroxybenzoate or derivatives thereof in the preparation of drugs for preventing and treating neurodegenerative diseases. Background technique [0002] Neurodegenerative diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), etc., are all associated with the loss of neurons or synapses . AD pathological studies have found that there is nerve regeneration in the brain of AD patients, but the effect is too weak to make up for the loss of neurons. Nerve regeneration in a broad sense includes two aspects: one is the regeneration of neuron cell bodies, that is, the division of neural stem cells to supplement the lost neurons. This phenomenon is called neurogenesis or neurogenesis (neurogenesis). AD can be treated by activating endogenous brain stem cells or transplanting exogenous stem cells to replenish dead or damaged n...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/216A61P25/00A61P25/28A61P25/16A61P25/14A61P25/08
Inventor 罗焕敏章正
Owner JINAN UNIVERSITY
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