The present invention will be described in further detail below in conjunction with specific embodiments and accompanying drawings.
 Take 103.4mg (0.287mmol) of triptolide and 1.75mg (0.01435mmol) of 4-dimethylaminopyridine and dissolve in 5mL of anhydrous dichloromethane, add 319.5mg (3.157mmol) of triethylamine, ice-bath to about 0°C, add 259.7 mg (2.87 mmol) of acryloyl chloride dropwise, gradually return to room temperature after dropping, stir for 2 hours, TLC detects that the reaction is complete, stop stirring, quench the reaction with saturated aqueous sodium bicarbonate, extract with dichloromethane, water The layer was extracted twice with dichloromethane, the dichloromethane extracts were combined, and most of the water in the dichloromethane extracts was washed away with saturated aqueous sodium chloride solution, then dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and prepared Separation on a thin-layer silica gel plate, using petroleum ether-ethyl acetate (2:1-1:1) as a developing solvent, gave 32.1 mg of a colorless transparent oil with a yield of about 27.0%. After detection, the structural formula of the compound is shown as formula I, ie triptolide acrylate.
 like Figure 1-3 Shown, formula I compound, molecular formula C 23 h 26 o 7 , ESI-MS m/z:414.1679[M+H] + (theoretical value).
 1 H NMR (600MHz, CDCl3) δ6.54(d, J=16.1Hz, 1H), 6.23(dd, J=17.3, 10.4Hz, 1H), 5.94(d, J=10.4Hz, 1H), 5.15(s ,1H),4.77–4.61(m,2H),3.84(d,J=3.2Hz,1H),3.52(dd,J=30.8,4.2Hz,2H),2.70(d,J=13.3Hz,1H) ,2.32(d,J=18.3Hz,1H),2.18(d,J=26.5Hz,2H),1.90(d,J=39.2Hz,2H),1.59(dd,J=4.8,16.0Hz,1H) ,1.24(m,1H),1.06(s,3H),0.97(d,J=7.0Hz,3H),0.85(d,J=6.9Hz,3H).
 13 C NMR (151MHz, CDCl3) δ173.24, 165.52, 159.98, 132.46 (CH 2 ), 127.78, 125.63, 71.13, 69.98 (CH 2 ),63.64,63.40,61.11,59.78,55.36,55.09,40.40,35.71,29.86(CH 2 ), 28.33, 23.47 (CH 2 ), 17.58, 17.08 (CH 2 ), 16.76, 13.74.
 1. Inhibition test of triptolide acrylate on subcutaneous transplanted tumor of HepG2 in nude mice
 1 Experimental materials
 1.1 Experimental animals: Nude mice, male, 4-6 weeks old, weighing about 18-20g, were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. (certificate number: 11400700270675).
 1.2 Cell: HepG2 cell line; drug: triptolide acrylate synthesized in the above example.
 1.3 Other experimental reagents and consumables:
Sterile normal saline, surgical scissors, tweezers, vernier caliper with display reading (Guangzhou Weijia Technology Co., Ltd.), 1mL syringe, cotton swab. RPMI-1640 medium, DMEM medium (Gbico, USA); fetal bovine serum (Gibco, North America); penicillin (double antibody), 0.25% trypsin (containing EDTA); apoptosis kit, cell cycle Kit (Hangzhou Lianke Biotechnology Co., Ltd.); cell culture flask, cell culture dish (Coring, New York, USA); 2mL cryopreservation tube (Coring, Los Angeles, USA); 96-well cell culture plate, 6-well cell culture plate (Coring ,Los Angeles, USA). RIPA lysate (strong), PMSF protease inhibitor, phosphatase protein compound inhibitor (Guangzhou Dingguo Biotechnology Company); Tween 20 (ST825, Biyuntian, Guangzhou Weijia Technology Co., Ltd.), SDS-PAGE gel reagent Box (Biyuntian, Guangzhou Weijia Technology Co., Ltd.), 5×LoadingBuffer (Biyuntian, Guangzhou Weijia Technology Co., Ltd.); Prism Protein Marker (Thermo, the United States); ECL chemiluminescent liquid (P0018A, Biyuntian, Guangzhou Weijia Technology Co., Ltd.) Technology Co., Ltd.); PVDF membrane (Biyuntian, Guangzhou Weijia Technology Co., Ltd.); thin filter paper, sponge, 1.5 μm sheet (Bio-Rad, the United States); methanol (Sinopharm Chemical Reagent Co., Ltd.); glycine (Qingdao Sheng Industrial Biotechnology Co., Ltd.), SDS (Beijing Baierdi Biotechnology Co., Ltd.), Tris Base (Shanghai Baiyan Biotechnology Co., Ltd.), TBS powder (Biyuntian, Guangzhou Weijia Technology Co., Ltd.); skimmed milk powder (BD, UK ).
 1.4 Experimental equipment
 One-ten-thousandth balance (Beijing Radolis Scientific Instrument Co., Ltd.), carbon dioxide incubator (Shanghai Boxun Industrial Co., Ltd.); cell ultra-clean workbench (Yisi Technology Co., Ltd., Singapore); low-speed desktop centrifuge (DT5- 3. Beijing Times Beili Centrifuge Co., Ltd.); microplate reader VICTORX5 (US, Perkinelmer); liquid nitrogen tank (LocatorPLUS, US). Multifunctional refrigerated centrifuge (5430R, eppendorf, China eppendorf Co., Ltd.), electrophoresis and membrane transfer device (Bio-Rad, the United States), speed-adjustable oscillator (HS260, IKA Shanghai Shengke Instrument Equipment Co., Ltd.), constant temperature metal bath (Q872), gel imaging system (XR+) (Bio-Rad, Shanghai Laboratories Co., Ltd.), shaker (SK-L330-Pro).
 2 Experimental process
 2.1 Establishment of animal models
 Establishment of HepG2-Luc cell line stably expressing luciferase: HepG2 cells in logarithmic growth phase were mixed with 1×10 5 /well into a 24-well plate and cultured overnight to allow the cells to fully adhere to the wall. Replace the original medium with 2 mL of fresh medium containing 6 μg/mL polybrene and add about 1×10 recombinant lentiviral particles stably expressing Luciferase. 5 For transfection units, incubate at 37°C for 4 hours and then add 2 mL of fresh medium to dilute polybrene. Continue culturing, replacing the medium containing the virus with fresh medium. Continue culturing, replace the medium containing puromycin for resistance screening, pick drug-resistant clones, continue screening for two weeks, and finally obtain the cell line HepG2-Luc that can stably express luciferase.
 Using the HepG2-Luc cell line to construct a mouse: resuscitate the frozen HepG2-Luc cells to 100cm 2 Culture flasks, cultured in vitro until logarithmic growth phase, digested with 0.25% trypsin containing EDTA, collected cells, centrifuged at room temperature 1000rpm for 3min, discarded supernatant, washed cells with serum-free DMEM medium, tested cells by trypan blue exclusion test Survival rate (the live cell ratio exceeds 95% to meet the experimental requirements). Add a little serum-free DMEM medium to resuspend the cells, and count the cells. Disinfect the back skin of nude mice with 75% alcohol, take 1×10 7 About 200 μL of the cell suspension was inoculated into the right anterior armpit of the nude mice, and kept for a week under sterile conditions, and observed whether there were subcutaneous tumors visible to the naked eye.
 Detect tumor growth using small animal in vivo imaging system: each nude mouse was intraperitoneally injected with 150 μL 30 mg/kg fluorescein substrate, observed for 15 minutes, anesthetized the nude mice with ether for 5 minutes in the induction box, and quickly transferred the anesthetized nude mice to the observation box , align and fix the head of the nude mouse in the conical nasal plug, set the parameters for fluorescence imaging (if there is a tumor growing, you can detect fluorescence signals with different intensities at the corresponding position), and transfer the nude mouse again after the imaging Go to the induction box and open the oxygen valve to resuscitate the nude mice.
 2.2 Animal grouping and administration
 Grouping: After 1 week, subcutaneous tumors were observed in all nude mice, and the growth of tumor pairs was verified by small animal in vivo imaging technology, so the nude mice were sorted and numbered according to body weight, and Excel software was used to generate 18 random numbers. Nude mice were divided into model group, low-dose group (100 μg/kg), middle-dose group (200 μg/kg) and high-dose group (400 μg/kg) according to the random number. After random grouping, the nude mice were weighed and the tumor volume was measured. Statistical tests were used to test the differences in nude mouse body weight and tumor volume among the groups.
 Administration: intraperitoneal injection, using a disposable sterile syringe for intraperitoneal injection of triptolide acrylate solution. The nude mice in the model group were given normal saline, and the treatment group was given triptolide acrylate solution according to the dose. Once a day, continuous administration for 13 days.
 2.3 Observation and recording
 Routinely observe the nude mice every day after treatment, including mental state, activity, diet, skin color and stool properties, etc., measure and record the body weight of the nude mice twice a week, measure the size of the transplanted tumor, and perform in vivo imaging once a week , monitor tumor growth and distant metastasis for 3 consecutive weeks until the end of administration.
 2.4 Materials
 After the administration, the animals were anesthetized, sacrificed by cervical dissection, the tumor body was completely stripped off, and the size of the tumor tissue was measured and recorded.
 2.5 Western Blot
 After the tumor tissue was thawed on ice, 50 mg was taken in a tissue homogenate tube, and 500 μL of tissue lysate containing protease inhibitors and phosphatase inhibitors was added (Roche brand tablet usage method: 1 tablet/10 mL). After high-speed homogenization, centrifuge at 12000rpm/min for 15min, take the supernatant, and measure the protein concentration of the sample by BCA method. After adjusting the concentration of each sample to be consistent, add protein loading buffer 5x Loading buffer, denature at 100°C for 10min, and store at -80°C. .
 Choose a suitable brand of 10% pre-mixed polyacrylamide gel preparation solution, and then prepare the gel used in the experiment according to the instructions (including separating gel and stacking gel, which takes about 2 hours); wait for the gel to solidify During the process, it is necessary to prepare the SDS-PAGE gel electrophoresis solution in advance, put the prepared gel in the electrophoresis tank, then add the electrophoresis solution, add 10-30 μL of sample to the sample tank of each well, and run the sample at a low voltage of 80V. After passing through the stacking gel, adjust the voltage to 100V so that the sample runs through the entire gel, and complete the SDS-PAGE gel electrophoresis operation; secondly, perform the membrane transfer operation (PVDF membrane, pre-wetted in methanol for 5 minutes in advance), the membrane transfer condition is : 300mA, 120-150min (the corresponding molecular weight is 100kDa boundary time selection, wherein the selected transfer time of the separated protein molecular weight less than 100kDa is 120min, and the selected transfer time of the protein molecular weight greater than 100kDa is 150min); Block with 5% skimmed milk for 2 hours and incubate the primary antibody overnight in the refrigerator at 4°C according to the corresponding band; wash the unbound primary antibody with TBST washing solution the next day (washing 5 times, 5 minutes each time); incubate the secondary antibody at 37°C 2h, the unbound secondary antibody was washed away with TBST washing solution (washing 5 times, 5 min each time); ECL luminescence solution was used as a substrate to expose the target band, and the analysis was recorded and the results were counted.
 3 Results and analysis
 3.1 In this experiment, a liver cancer cell line HepG2-Luc capable of stably expressing luciferase was established, and a nude mouse subcutaneous liver cancer implantation model was constructed using this kind of cells.