A kind of spiro sesquiterpene dimer compound and preparation method thereof
A sesquiterpene and dimer technology, which is applied in the field of spirocyclic sesquiterpene dimer compounds and their preparation, can solve problems such as unseen and unseen compounds related reports and the like, and achieve good anti-tumor effect
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Embodiment 1
[0062] Example 1 Preparation of the compound of the present invention
[0063] 1. Experimental materials:
[0064] ①Medicinal materials
[0065] Huadong blue thorn head root was purchased from Anguo Pharmaceutical City, Hebei Province in June 2017, and was identified as the dry root of Echinops grijsii Hance (Echinops grijsii Hance) by the inventor of the patent, Dr. Liu Tingting.
[0066] ②Reagents and fillers
[0067] Column chromatography silica gel, 200-300 mesh (reagent grade), purchased from Qingdao Ocean Silica Gel Desiccant Factory;
[0068] Thin layer chromatography silica gel GF254 (chemically pure), purchased from Qingdao Ocean Silica Gel Desiccant Factory;
[0069] Sephadex LH-20 dextran gel, purchased from Amersham, Sweden;
[0070] GF254 silica gel preparation thin layer, purchased from Yantai Jiangyou Silicone Development Co., Ltd.;
[0071] Analytical reagents such as petroleum ether, n-hexane, chloroform, ethyl acetate, acetone, methanol, etc. were purchased from Beijing Ch...
Embodiment 2
[0091] Example 2 In vitro anti-tumor experiment
[0092] 1. Experimental cells
[0093] Human liver cancer cells (HepG2), human cervical cancer cells (Hela) and human laryngeal cancer cells (Hep2) were purchased from the Central Laboratory of Peking Union Medical College Hospital, Chinese Academy of Medical Sciences.
[0094] 2. Experimental drugs
[0095] The spirocyclic sesquiterpene dimer (cyanotopein A) prepared in Example 1 and camptothecin (purchased from Sinopharm Chemical Reagent Co., Ltd.) were used as positive control drugs. Prepare a drug solution with a concentration of 5-80μM in the culture medium.
[0096] 3. Determination of drug anti-cancer activity (MTT method)
[0097] Human liver cancer cells (HepG2), human cervical cancer cells (Hela) and human laryngeal cancer cells (Hep2) were cultured using high-sugar DMEM medium containing 10% fetal bovine serum. Then the three types of cells in the logarithmic growth phase are calculated according to 2×10 4 The density of cells...
Embodiment 3
[0105] Example 3 Anti-inflammatory activity-impact on LPS-induced mouse macrophages to secrete inflammatory factors
[0106] 1. Experimental cells
[0107] The mouse macrophage cell line RAW264.7 was purchased from Union Cell Resource Center of Chinese Academy of Sciences.
[0108] 2. Experimental drugs and reagents
[0109] Trypsin, dimethyl sulfoxide (DMSO) (Gibco); RP-MI1640 cell culture medium (Hyclone); Nitric oxide (NO) test kit (Biyuntian); MTS reagent (Promega); LPS (Sigma), other reagents are commercially available analytical grade.
[0110] 3. Cell culture
[0111] RAW264.7 mouse mononuclear macrophages were cultured in RP-MI 1640 medium containing 10% fetal bovine serum, penicillin (1×105U / L) and streptomycin (100mg / L) at 37℃, 5% CO 2 Cultivate in an incubator and pass the next day.
[0112] 4. Cytotoxicity test
[0113] Take the prepared mouse RAW264.7 cells to 5×10 4 Cells / wells are placed in 96-well culture plates at 37℃, 5% CO 2 After culturing for 24 hours, add different c...
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