Application of lobeline in preparing medicament for treating apoptosis of the nerve cell

A neuron and lobeline technology, applied in the field of biomedicine, can solve problems such as the central nervous system of lobeline that have not been reported

Inactive Publication Date: 2009-01-14
上海国联干细胞技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Clinically, it is mainly used for neonatal asphyxia, asphyxia caused by carbon monoxide poisoning, poisoning of inhaled anesthetics and other central depressants (such as opioids, barbiturates), and respiratory failure caused by infectious diseases such as pneumonia and diphtheria, but has not been reported yet. Lobeline for the treatment of degenerative diseases of the central nervous system (CNS)

Method used

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  • Application of lobeline in preparing medicament for treating apoptosis of the nerve cell
  • Application of lobeline in preparing medicament for treating apoptosis of the nerve cell
  • Application of lobeline in preparing medicament for treating apoptosis of the nerve cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 The effect of lobeline on the uptake of dopamine by D8 cells

[0059] 1. Establishment of in vitro screening model

[0060] Establishment of activity-screening cell lines targeting dopamine neurotransporter (DAT). By cloning the full-length coding sequence of rat DAT (GeneBank accession number: GI31000978) into the multiple cloning site of the pCDNA3 vector (Invitrogen, USA), transfection into Chinese hamster ovary cells (CHO) by electroporation, 48 hours Then cultured in 1640 medium containing G418. After 10 days, all the cells in the control group died, while many cell clones formed in the experimental group. After the clones were picked and cultured for one week, when the cells covered the bottom of the well, the medium was sucked off, and digested with trypsin as above. Cells in each well were seeded in corresponding wells of two 96-well plates. After confluency, one of the plates was used for isotopic flux measurements. The cells in the wells with hi...

Embodiment 2

[0064] Example 2 Lobeline inhibits the uptake of dopamine by rat striatal synaptosomes

[0065] Preparation of synaptosomes: Adult male SD rats were killed by decapitation and the striatum was isolated and rinsed with ice-cold 0.32M sucrose (dissolved in 1M phosphate buffer). Homogenize the solution with a glass homogenizer at 4°C for 10-12°C, centrifuge the homogenate at 1100g for 10 minutes at 4°C, and take the supernatant and centrifuge at 10,000g for 20 minutes at 4°C. The pellet was resuspended in artificial cerebrospinal fluid (103mM NaCl, 1mM CaCl2, 1mM KH2PO4, 25mM HEPES, 1mM MgCl2, 27mM NaHCO3, 5.4mM glucose, pH7.4, 1mM ascorbic acid, 0.01mM pargyline) for later use.

[0066] When experimenting with synaptosomes, different concentrations of lobeline hydrochloride and other compounds were first mixed with the same amount of synaptosomes for 5 minutes, then added with a final concentration of 0.1 μM tritium-labeled dopamine, placed at 37 ° C for 10 minutes, and passed t...

Embodiment 3

[0068] Example 3 The effect of lobeline on the reverse transport ability of DAT

[0069] D8 cells were cultured in a 48-well plate (Costar) until the plate was confluent (approximately 60,000 cells per well). Discard the culture medium, wash once with PBS, suck off the PBS solution, add 90ul HBS (10mM Hepes, 100mM NaCl, pH8.0) to each well, 10ul 3 H-DA (Amersham Pharmacia Biotech), 100 μM vitamin C and 100 μM parjiline, incubated at 25°C for 10 minutes; washed 3 times with PBS, added 90ul HBS to each well, 10ul drugs of different concentrations, incubated at 25°C for 20 minutes, pipetted The supernatant of each well was added to 1.2ml of scintillation fluid, and put into a liquid scintillation counter (Beckman LS 5000TA) to detect the content of the isotope (DMP value) to measure the effect of the drug on the transport activity of the dopamine transporter .

[0070] RESULTS: Lobeline did not significantly affect DAT-mediated dopamine reverse transport at the dose used to inh...

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Abstract

The invention is designed to provide lobeline applied for preparing medicine used for curing neuron apoptosis. The invention also relates to the applications of the lobeline for curing a CNS degenerative disease medicine. The lobeline up-regulates the concentration of synapse intermission DA by inhibiting the transport of DAT, so the apoptosis speed of a nerve cell can be stopped or retarded with little adverse effect. The lobeline is proved that the lobeline can protect dopaminergic neuron, so the invention provides the new medicine used for the clinical treatment of the CNS degenerative disease.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the application of lobeline in the preparation of medicines for treating neuron apoptosis diseases. Background technique [0002] Neurons, also called nerve cells, are the basic units that make up the structure of the nervous system. Neurons are cells with long processes, which consist of a cell body and cell processes. Cell bodies are located in the brain, spinal cord, and ganglia, and cell processes can extend to various organs and tissues throughout the body. Neurons are very different from other cells in the body: Mature neurons neither move nor come together and divide normally. If a mature neuron dies, (except very rarely) it is not replaced by a new neuron. [0003] The nervous system is the dominant system in the body. Various information of the internal and external environment, after being received by the receptors, is transmitted to the central centers at all levels of t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/445A61K9/00A61K9/08A61K9/107A61K9/14A61K9/16A61K9/20A61K9/48A61P25/00A61P25/14A61P25/16A61P25/28
Inventor 郭礼和暨荀鹤栗超跃
Owner 上海国联干细胞技术有限公司
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