Triterpenoid compounds, and preparation and application thereof
A compound and technology of triterpenoids, applied in the field of structure and preparation of triterpenoids, to achieve the effect of simple preparation method, high purity and optimized structure
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Embodiment 1
[0034] Example 1 Preparation of Compound A and Compound B
[0035] A preparation method for obtaining compounds A and B with significant NGF-mimicking activity from Motherwort Motherwort, the specific steps are:
[0036] (1) Grinding and extraction: After 300 g of Motherwort, a traditional Chinese medicine, was crushed, it was extracted with 1L of methanol (industrial grade) at room temperature for 5 days (shaking). After suction filtration and concentration, a crude sample of the methanol extract was obtained, and the solvent was partitioned with water and ethyl acetate to obtain a total of 6 grams of a crude sample of the ethyl acetate layer and 14 grams of a crude sample of the aqueous layer.
[0037](2) Separation and purification: separate the crude ethyl acetate layer through a silica gel open column (200-300 mesh, the following is the solvent volume ratio, n-hexane:chloroform=2:8,1:9,0:10, chloroform : Methanol=98:2, 96:4, 94:6, 92:8, 90:10), collect and combine chlo...
Embodiment 2
[0041] Example 2 Qualitative identification of the physical and chemical characteristics and compound structure of compound A, B obtained in embodiment 1:
[0042] The chemical structures of compounds A and B were determined by mass spectrometry (including high-resolution mass spectrometry) and nuclear magnetic resonance.
[0043] Compound A, a colorless solid, is easily soluble in methanol. When analyzed by TLC, using chloroform:methanol (8:2) as the developing solvent, the Rf value is 0.5 on a forward silica plate with a height of 5 cm. This sample has significant UV absorption at 228 nm.
[0044] Compound B, a colorless solid, is easily soluble in methanol. When analyzed by TLC, using chloroform:methanol (9:1) as developing solvent, on a forward silica plate with a height of 5 cm, the Rf value is 0.5. This sample has significant UV absorption at 228 nm.
[0045] Compound A, the molecular formula is C 36 h 56 o 11 ; HR FT-ICR MS: m / z 687.3707[M+Na] + , theoretical ...
Embodiment 3
[0047] Example 3 The compound exhibits significant NGF-mimicking activity in PC12 cells, an in vitro screening model of senile dementia.
[0048] experiment method:
[0049] 1) Culture of PC12 cells: inoculate 20×10 4 Each cell was placed in a 100 mm culture dish, containing 10 ml of DMEM medium (containing 10% horse serum, 5% fetal bovine serum), and the medium was replaced every two days, and subculture was carried out when the cell density reached the requirement. First wash the cells twice with PBS, then add 10 ml PBS to the culture dish, at 37 ℃, 5% CO 2 Let it stand in the incubator for 10 minutes, take it out and wash it, transfer the PBS with suspended cells to a 15 ml disposable centrifuge tube, count with a hemocytometer after centrifugation, and add 1 ml of serum to each well of the 24-well plate DMEM medium, after cell counting, each well was inoculated with 2×10 4 cells, mixed well, in CO 2 Samples were added after 24 hours in the incubator.
[0050] 2) A...
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