Application of polyketide in preparation of angiogenesis inhibiting medicine
A technology of polyketides and angiogenesis, applied in the separation/purification of active ingredients of heterocyclic compounds, carbonyl compounds, drug combination, etc., can solve problems that have not been reported
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Embodiment 1
[0029] Example 1 Preparation of polyketides
[0030] (1) At room temperature, extract the dry powder of C. chinensis with 95% ethanol by percolation, and then evaporate the ethanol to dryness in vacuum to obtain the crude extract of C. cicadae;
[0031] (2) Disperse the above crude extract with water, then extract with chloroform, and evaporate the chloroform to dryness in vacuo to obtain the chloroform extract;
[0032] (3) The chloroform extract was subjected to silica gel column chromatography, and gradient elution was carried out with petroleum ether-ethyl acetate (100:0-5:1), and the eluent of petroleum ether-ethyl acetate (7:3) was collected. The solvent was evaporated to dryness in vacuo to obtain the target crude product;
[0033] (4) The above crude product was subjected to silica gel column chromatography and eluted repeatedly with petroleum ether-ethyl acetate (7:3) to obtain the target product.
[0034] The chemical structures and names of the polyketides 1, 2, 3...
Embodiment 2
[0040] Example 2 Effects of polyketides 2, 3, 4 and 5 on the proliferation of human umbilical vein endothelial cells (HUVEC)
[0041] Purpose and principle: CCK-8 method was used to detect the proliferation of HUVEC, which has been widely used in cell proliferation assay research. CCK-8 reagent contains WST-8 (2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfonic acid benzene) -2H-tetrazolium monosodium salt), which is reduced by the dehydrogenase in the mitochondria of the cell under the action of the electron carrier 1-Methoxy PMS (1-methoxy-5-methylphenazine dimethyl sulfate) to have Highly water-soluble yellow formazan product (Formazan). The amount of formazan produced is directly proportional to the number of viable cells. The absorbance was measured at a wavelength of 450nm by an enzyme-linked immunosorbent detector, which can indirectly reflect the number of living cells.
[0042] Methods: HUVEC cells were seeded in 96-well plates at 5000 cells per well, a...
Embodiment 3
[0047] Example 3 Polyketide 4 and 5 Compounds Inhibit HUVEC Capillary Formation Experiment
[0048] Purpose and principle: In vitro angiogenesis assay kit was used to investigate capillary formation in HUVEC. Matrigel is a soluble basement membrane matrix extracted from mouse EHS sarcoma. The main components include: laminin, collagen IV, etc., and also contain TGF-β fibroblast growth factor, tissue plasminogen activator and other growth factors naturally expressed in EHS tumors. At room temperature, Matrigel self-aggregates to produce a bioactive matrix material similar to the basement membrane of mammalian cells. HUVEC can adhere to form tubes on Matrigel, which can be used to study the effect of drugs on capillary formation by endothelial cells.
[0049] Methods: All operations involving Matrigel were carried out on ice, and the orifice plates and pipette tips used had to be pre-cooled. Add 50 μl of Matrigel gel solution into a pre-cooled 96-well plate, and place it in a...
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