Oleanane triterpene saponin compounds which are effective on treatment of dementia and mild cognitive impairment(MCI), and improvement of cognitive function
A technology of mild cognitive impairment and triterpene saponins, applied in medical preparations containing active ingredients, drug combinations, organic active ingredients, etc.
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Embodiment 1
[0037] Embodiment 1: Extraction of oleanane type triterpene saponins
[0038] Oleanolic acid saponins and ivy saponins were isolated from the following herbal medicines known to contain a large amount of oleanolic acid saponins and ivy saponins according to the following method.
[0039] Every 1kg (dry weight) of Aralia elata, Pulsatilla chinensis, Lonicera japonica and Kalopanax pictus were extracted with 7L of 50% ethanol for 4 hours under reflux. This step was repeated 2 times. The extract was filtered and concentrated under reduced pressure at 50 °C using a rotary evaporator. Water was added to the obtained concentrate to about 5 times the volume (V / W) of the volume. The resulting suspension was mixed with an equal volume of water-saturated n-butanol, placed in a separatory funnel, shaken and left for 24 hours. The upper butanol layer was separated. After 2-3 more separations, the butanol fraction was concentrated using an evaporator and the solvent was completely remo...
Embodiment 2
[0041] Example 2: Separation and structural analysis of oleanane-type triterpene saponins
[0042] The methanol fraction was subjected to high performance liquid chromatography (HPLC) chromatography using an acetonitrile / water mixture solvent. Using a PDA detector, select the main peak with the maximum absorption value at 210 nm, which represents a specific triterpene absorption spectrum. Selected peaks were then isolated by preparative HPLC using acetonitrile / water mixture solvent at a flow rate of 9.5 mL / min. After concentration using an evaporator, the solvent was completely removed by drying in a vacuum oven. Component analysis was performed at 210 nm using a YMC J'Sphere ODS-H80 column. in 2N H 2 SO 4 Under acidic conditions, heat at 100°C for 60 minutes to hydrolyze the components of the separated peaks and remove the sugars of saponins. The resulting product was analyzed by HPLC together with oleanolic acid and helexinin standard substances purchased from Sigma. C...
Embodiment 3
[0047] Example 3: Passive Avoidance Test
[0048] A dark avoidance test was carried out to determine the memory-improving effects of the above-mentioned 8 kinds of oleanolic acid saponins or hedera saponins in vivo. In addition, the saponin compounds aglycon, oleanolic acid and helexin were included in the test.
[0049] A shuttle box measuring 50 cm x 15 cm x 40 cm was used. The tank is divided into two compartments by a gate. One compartment was brightly lit, the other was covered with a black cloth.
[0050] Place mice in brightly lit compartments. Then, when the gate was opened, the mice entered the darkened compartment within 20 seconds, as mice prefer the dark. In this way the waiting time, or the time elapsed until the mouse leaves the light compartment and enters the dark compartment, is determined. On the first day, this training test was performed until all mice entered the dark compartment within 20 seconds.
[0051] The next day, all trained mice were placed ...
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