15-oxospiramilactone derivatives, and preparation method and applications thereof
A kind of technology of meadowfoam and derivatives, applied in the field of medicine
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0047] Preparation of compounds of the present invention:
[0048] (1) The preparation routes of compounds 1-9, 25, and 26 are:
[0049]
[0050] Preparation:
[0051] Compound 1: Take the dry compound 15-oxospirandrolactone (S-3) (2.00g, 6.06mmol) from Meadowsweet pink, in N 2 Dissolve with 5ml of dry dichloromethane under protection, inject oxalyl chloride (7.28mmol, 692ul) into a long-necked low-temperature reaction bottle filled with 5ml of dry dichloromethane under nitrogen protection, and stir at -78°C for 20 minutes, then Slowly add DMSO (14.56mmol, 1032ul, in5ml DCM) solution dropwise, after 5 minutes dropwise, stir at low temperature for 30 minutes, then slowly add the dichloromethane solution of compound S-3 into the reaction flask, at -78 After stirring for 1 hour under stirring at ℃, slowly add triethylamine (14.56mmol, 2020ul) dropwise under rapid stirring. Extracted with methyl chloride, combined with dichloromethane, washed with saturated brine, dried over...
Embodiment 2
[0175] The inhibitory effect of the compound of the present invention on various tumor cell lines:
[0176] (1) Experimental method
[0177] 1. Inoculate cells: Use culture medium (DMEM or RMPI1640) containing 10% fetal bovine serum to prepare a single cell suspension, inoculate 5000-10000 cells per well into a 96-well plate with a volume of 100 μl per well, and adherent cells in advance 12 hour inoculation.
[0178] 2. Add the compound solution to be tested (fixed concentration of 40 μM for primary screening, at which concentration the compound that inhibits the growth of tumor cells near 50% is set to enter the gradient rescreening at 5 concentrations), the final volume of each well is 200 μl, and each treatment is set at 3 multiple holes.
[0179] 3. Color development: After incubating at 37 degrees Celsius for 48 hours, add 20 μl of MTT solution to each well. Continue to incubate for 4 hours, terminate the culture, discard the culture supernatant in the well, add 200 μl...
Embodiment 3
[0187] The inhibitory effect of the compounds of the present invention on the Wnt signaling pathway reporter gene Top-flash:
[0188] (1) Experimental method:
[0189] 1) Preparation of cell culture and conditioned medium:
[0190] HEK293T cells were cultured in DMEM (Invitrogen) containing 10% fetal bovine serum at 37°C, CO 2 Concentration 5%. Subculture every other day. Maintain a density of 70-80% when subcultured. The L cell line stably secreting mouse Wnt3a protein (CRL-2647 was purchased from the US ATCC cell bank) and the control strain (CRL-2648 was purchased from the US ATCC cell bank), maintained at 50ug / ml G418 and 10% fetal bovine serum DMEM, 37°C, CO 2 At a concentration of 5%. When the cells grew to about 70% density, the medium was changed (DMEM containing 10% fetal bovine serum). After four days of continuous culture, the culture medium was collected and centrifuged, and the supernatant was retained to obtain the Wnt-conditioned medium. After titer deter...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More