New type natural active compound for anti tumor, preparation method, and application
An anti-tumor activity and compound technology, applied in the field of medicine, can solve the problems of unresearched tumors, unclear active ingredients, etc., and achieve the effects of good anti-multidrug resistance, simple, safe and easy preparation method and strong activity.
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Embodiment 1
[0021] 30Kg (dry weight) of the whole plant of Myrica rubra is crushed and extracted with 95% ethanol (50L × 3 times), concentrated under reduced pressure to obtain 1680g of total extract. The total extract was dissolved in 3.0L of warm water at about 40-60°C, cooled, extracted with chloroform (1.0L x 10 times), concentrated under reduced pressure to obtain 250g of chloroform partial extract. Part of the chloroform extract was subjected to silica gel column chromatography, petroleum ether (60-90°C)-acetone (petroleum ether, 50:1, 20:1, 10:1, 5:1, 2:1, acetone) gradient elution, It was divided into 9 parts (TLC was used for detection, and the same parts were combined together); Fr.4 was prepared with a thin layer of silica gel, and the developing solvent was petroleum ether: ethyl acetate (4:1, V / V), and 1250 mg of the compound was obtained by separation and purification.
[0022] Compound 1 is an orange-red oil, which reacts with 10% ethanol sulfate on a TLC silica gel thin-la...
Embodiment 2
[0034] Embodiment 2: anti-inflammatory effect of the present invention
[0035] According to the pharmacy method, the extract of the present invention can be used for preparation as medicine for enhancing body immunity or anti-inflammation.
[0036] The following pharmacological experiments show that the extract of the present invention has pharmacological activities such as enhancing immunity and anti-inflammation: 40 mice, half male and half female, with a body weight of 80-150 g, were subcutaneously injected with 0.5% 2,4-ONFB ethanol solution 100 pl via the back As an antigen to sensitize it, the sensitization was established 4 days after the injection of 2,4-DNFB, and on the 6th day, 1% 2,4-DNFB-olive oil solution was applied to the right ear of the mouse as an antigen challenge, so that the mouse developed ear fringe Contact dermatitis. They were randomly divided into 4 groups, and the bayberry extract was orally administered for 7 consecutive days from the day before t...
Embodiment 3
[0037] Embodiment 3: antitumor effect of the present invention
[0038] The survival rate of cells was detected by SRB method, and the apoptosis of cells was detected by Hoechst33258 nuclear staining method, agarose gel electrophoresis method and flow cytometry to evaluate the anti-tumor activity of compound 1.
[0039] 1. Reagent DMEM and calf serum were purchased from GIBCO BRL Company, ethidium bromide (EB) was purchased from Shanghai Sangon Company, Hoechst33258 was purchased from Sigma Company, Proteinase K (Proteinase K), 100bp DNA ladder was purchased from NEB Company, RNA Enzyme A (RNaseA) was purchased from PROMEGA Company, pyridinium iodide (PI) was purchased from Merck Company, agarose (Argarose) was purchased from BioWest Company in Spain, and other reagents were of domestic analytical grade.
[0040] 2. Instrument CO 2 Incubator, inverted microscope, fluorescence microscope, high-speed refrigerated centrifuge, horizontal electrophoresis device, gel scanning analy...
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