Application of aminoadamantane mononitrate compound in preparation of drugs for preventing and treating diseases
A technology of aminoadamantane and compounds, applied in sensory diseases, drug combinations, respiratory diseases, etc.
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Embodiment 1
[0091] Example 1. The protective effect of aminoadamantane nitrate compounds on cerebellar granule cells.
[0092] Primary isolated neonatal rat cerebellar granule cells were divided into 1.2×10 5 Each well was seeded in a 96-well plate and cultured in BME medium containing 10% FBS+25mM KCl+2mM Glutamine+1% double antibody. After 24 hours, cytarabine at a final concentration of 10 μM was added to inhibit the proliferation of glial cells. From the 4th day, add glucose with a final concentration of 5mM every 4 days to supplement the energy metabolism and water evaporation of the cells, and place them in a cell culture incubator (37°C, 5% CO 2 ) for 10 days. Primary cerebellar granule cells were divided into normal control group, glutamate group, different aminoadamantane nitrate compound pretreatment group and memantine pretreatment control group induced by 200 μM glutamate. Add different concentrations of compounds MN-04, MN-05, MN-06, MN-07, MN-08, MN-09 and memantine (mema...
Embodiment 2
[0093] Example 2. Inhibition of NMDA receptors by aminoadamantane nitrate compounds.
[0094] The inhibitory activity of aminoadamantane nitrate compounds on NMDA receptors in HEK293 cells stably transfected with human NR1 and NR2A was studied by patch clamp technique. We recorded changes in NMDA receptor channel currents using the whole-cell patch clamp technique. The clamping voltage of HEK293 cells was set to -70mV, adding various concentrations of test compounds and L-glutamate solution to stimulate the opening of NMDA channels, and measuring the inhibitory effect of each compound on the increase of L-glutamate-induced current, and then calculating the effect of each compound on NMDA channel The half inhibitory concentration constant IC 50 .
Embodiment 3
[0095] Example 3. Time-concentration curve of nitric oxide in plasma after intravenous administration of MN-08 to rats.
[0096] Eight SD rats weighing 250±20 g were used and randomly divided into two groups, with 4 rats in each group. MN-08 (25 mg / kg) or ISDN (3 mg / kg) was injected into the tail vein, and about 400 μL of blood (using EDTA- K2 anticoagulant), and centrifuged at 3000r / min for 10min to get the supernatant, and stored the plasma sample at -20°C for testing. The NO content in plasma was determined according to the instructions of Beyontien's total nitric oxide detection kit.
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