Application of cholest-4-ene-3,6-dione in preparing drug for treating or preventing neuron injury

A technique for neuron damage and cholesteric, applied in drug combinations, nervous system diseases, neuromuscular system diseases, etc.

Inactive Publication Date: 2018-05-18
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Application of cholest-4-ene-3,6-dione in preparing drug for treating or preventing neuron injury
  • Application of cholest-4-ene-3,6-dione in preparing drug for treating or preventing neuron injury
  • Application of cholest-4-ene-3,6-dione in preparing drug for treating or preventing neuron injury

Examples

Experimental program
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Embodiment 1

[0024] Example 1. Glutamate-induced HT-22 oxidative stress injury model

[0025] Take the HT-22 cells that have been passaged three times in a medium dish after resuscitating with liquid nitrogen. Before use, observe that the cells are in good condition, the cell bodies are round and full, the synapses are complete, and there are almost no dead cells. Digest with 0.5ml of 0.25% trypsin for half a minute , blown into a single-cell suspension, collected in a 15ml centrifuge tube, added complete medium to 3ml, centrifuged at 1000rpm for 3min, removed the supernatant, and evenly dispersed the precipitate in 4ml complete DMEM medium (DMEM+10%FBS+1%P / S), take 0.8ml of cell suspension for passage, then take an appropriate amount of suspension and add it to complete DMEM medium, dilute to 4*10 4 cells / ml, planted in a 48-well plate (200 μl / well), and used for experiments after 24 hours.

[0026] First, cholest-4-ene-3,6-dione was dissolved in DMSO to make a final concentration of 10...

Embodiment 2

[0032] Example 2. Apoptotic injury model of cerebellar granule neurons induced by potassium deprivation

[0033] Take out 7-day-old rats, cut off their heads with major surgery, and use small ophthalmological scissors to cut from the vertebral foramen to the upper edges of the left and right ears, then use tweezers to open the upper layer of the brain to expose the brain tissue, take out the cerebellum tissue and put it in a container. Place DMEM without phenol red (for dissection fluid) in a medium dish on an ice pack.

[0034] After removing the meninges and blood vessels on the cerebellum with ophthalmic forceps, use tissue scissors to mince the cerebellar tissue. Transfer the tissue into trypsin with a concentration of 0.25%, digest at 37°C for 15 minutes, add complete BME medium to 15ml, then add 100μl of DNase I (8mg / ml), blow off the tissue mass with a dropper, and then Centrifuge at 1000rpm for three seconds, transfer the supernatant to another centrifuge tube, centri...

Embodiment 3

[0043] Example 3. Excitotoxicity model of cerebellar granule neurons induced by glutamate

[0044] Take the cerebellar granule neurons that have been cultured for 7 days, observe under the microscope, the cells are round, the synapses are obvious, and the distribution is even, that is, it can be used for experiments. Do the following group processing:

[0045] a) Control group: pre-applied without Mg 2+ Locke,s Buffer

[0046] b) Glutamate group: no Mg pre-applied 2+ Locke's Buffer+200μM Glu

[0047] c) MK801 group: pre-coated with 10 μM MK801 without Mg 2+ Locke's Buffer+200μM Glu

[0048] d) Cholester-4-ene-3,6-dione group: pre-coated with 10 μM steroid without Mg 2+ Locke's Buffer+200μM Glu

[0049] Take the cerebellar granule neurons cultured for 7 days, collect the BME-conditioned medium of each well, and use Mg-free 2+ Locke, sBuffer solution washed cells three times, and then Mg-free2+ The drug solution prepared in Locke's Buffer solution was pre-coated at 37°C ...

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Abstract

The invention discloses an application of cholest-4-ene-3,6-dione in preparing a drug for treating or preventing neuron injury. Cholest-4-ene-3,6-dione can relieve hippocampus neuron oxidative stressinjury caused by glutamic acid, cerebellar granule neuron apoptosis injury caused by potassium deprivation and cortical neuron and cerebellar granule neuron oxidative stress injury caused by glutamicacid, also has a function of remarkably reducing infarct volume in an MACO (middle cerebral artery occlusion) model of a stroke rat and has the potential of a neuroprotective agent used for being developed into multiple acting mechanisms. Neurodegenerative diseases such as the Parkinson's disease, the Alzheimer's disease, the tau protein disease, amyotrophic lateral sclerosis and the like as wellas cerebral stroke, cerebral injury and spinal cord injury are all caused by neuron injury, and neuron injury is further aggravated. Cholest-4-ene-3,6-dione has a protection function on neuron injuryand has potential functions in drugs for treating or preventing neuron injury.

Description

technical field [0001] The invention relates to the application of cholest-4-ene-3,6-dione in the preparation of medicines for treating or preventing neuron damage. Background technique [0002] Cholester-4-ene-3,6-dione is a cholesterol derivative with the following structural formula: [0003] [0004] At present, the synthesis methods of this compound mainly include cholesterol as raw material (1) obtained by one-step oxidation reaction of Jones reagent (Zhang Wei, et al, Steroids, 2014, 86, 39-44); (2) obtained by PCC / dichloromethane Obtained by one-step oxidation reaction (Huang, Yanmin et al, Faming Zhuanli Shenqing, 103044514); (3) obtained by one-step oxidation reaction of PDC / DMF (Hector Markus et al, Synthetic Communications, 26(6), 1075-82; 1996); (4) Obtained through one-step oxidation reaction of tetrapropylammonium perruthenate / N-methyl-N-morpholine oxide / dichloromethane (Moreno, M.J.S.et al, Tetrahedron Letters, 32(27), 3201-4; 1991 ); (5) obtained throug...

Claims

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Application Information

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IPC IPC(8): A61K31/575A61P25/00A61P25/16A61P25/28A61P21/00A61P9/10
CPCA61K31/575
Inventor 张静夏颜光美银巍陈文礼张伟李心花
Owner SUN YAT SEN UNIV
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