| Objective(s):The compound C18H17NO6 was prepared into nanoliposomes to solve the problem of insoluble in water and change the type of preparation.Through the orthogonal experiment,the best ratio scheme of liposome was obtained,and the factors affecting the drug loading amount were explored,and the preparation scheme was adjusted.The dose-response relationship of C18H17NO6 nanoliposomes against tumor in vitro was studied.The in vitro combined action and intensity of C18H17NO6 nanoliposomes combined with cisplatin on human cancer cell lines were observed.The in vivo anti-tumor effect of C18H17NO6 nanoliposomes was observed by human cancer xenograft model.The effect of C18H17NO6 nanoliposomes on apoptosis of cancer cells was investigated by flow cytometry.Methods:1.Nano-liposomes were prepared by thin film dispersion-ultrasonic method.The factors affecting the drug loading of liposomes were determined by orthogonal experiments.The amount of phospholipids and cholesterol was adjusted.The preparation method of nano-liposomes was adjusted,and the control particles were filtered by 220nm sterile filter.The diameter method was used to determine the drug loading of nanoliposomes by HPLC.2.In vitro anti-cancer experiment,and different concentrations of C18H17NO6 nanoliposomes were detected by MTT assay on HCT-116,HepG2,andXWLC-05,MCF-7 and A549 inhibited proliferation in vitro.3.In vitro combined with cisplatin assay,MTT assay was used to detect the proliferation inhibition effect of different concentrations of C18H17NO6 nanoliposomes on MCF-7 and XWLC-05.The combined action of C18H17NO6 nanoliposomes and clinical antitumor drugs was calculated by the formula of Jin’s formula.4.Using the human cancer xenograft model,the tumor relative volume was measured by measuring the volume of tumor in nude mice,and the anti-tumor effect of C C18H17NO6 nanoliposomes in vivo was observed.After the end of the experiment,the body weight and the weight of the main organs were accurately weighed,and the safety of the drug was initially evaluated.5.Using flow cytometry to explore whether C18H17NO6 nanoliposomes induce tumor cell apoptosis.Results:1.The compound C18H17NO6 can be prepared as a nano-liposome,and the amount of soybean phospholipid and cholesterol is 5:1,and the liposome-loaded amount is 1.26 mg/ml and the particle diameter is less than 220 nm by HPLC.The direct analysis method of orthogonal analysis showed that the amount of soybean phospholipids and cholesterol was the main influencing factor.The more the fixed ratio of soybean phospholipids to cholesterol,the greater the drug loading of nanoliposomes.2.C18H17NO6 nano-liposomes in vitro experiments have demonstrated its ability to inhibit a variety of tumor cells in vitro,and different sensitivity to different tumor cells.Inhibition of HCT-116 under different concentrations of compound C18H17NO6 nanoliposomes 0.5 μg/mL,1 μg/mL,2 μg/mL,3 μg/mL,4 μg/mL The rates were 51.28%,73.38%,90.09%,98.28%,and 99.95%,respectively,IC50 is 0.86μg/mL.The inhibition rates of HepG2 were 30.87%,32.18%,46.32%,78.34%,and 96.91%,respectively,IC50 is 1.86 μg/mL.The inhibition rates of XWLC-05 were-2.49%,1.04%,27.78%,78.12%,and 85.59%,respectively,IC50 is 2.48 μg/mL.The inhibition rates for MCF-7 were-9.85%,-2.63%,17.57%,46.38%,and 61.13%,respectively,IC50 is 3.44 μg/mL.The inhibition rates of A549 were 19.12%,23.54%,32.27%,39.67%,and 66.52%,respectively,IC50 is 3.17 μg/mL.Among them,C18H17NO6 nanoliposomes were most sensitive to HCT-116 cell line.At the maximum dose concentration of 4 μg/mL,the maximum inhibition rate of tumor cells was 99.95%.Moreover,the anti-tumor effect of C18H17NO6 nano-liposomes has a significant dose-effect relationship.The greater the concentration of in vitro injection,the greater the inhibitory effect on tumor cells.3.C18H17NO6 nano-liposomes combined with cisplatin on XWLC-05,MCF-7 in vitro anti-tumor cell research,when the concentration of nano-liposomes is 0.5 μg/mL and 1 μg/mL,C18H17NO6 nanoliposomes combined with cisplatin antagonizedXWLC-05 with Q values of 0.72 and 0.78 and nanoliposomes at 2 μg/mL,C18H17NO6 nanoliposomes combined with cisplatin showed an additive effect on XWLC-05 with a Q value of 0.93.C18H17NO6 nano-liposomes combined with cisplatin showed an additive effect on MCF-7 in vitro,and the concentration of nano-liposomes was 0.5 μg/mL,1 μg/mL,2 μg/In the case of mL,the Q values are 0.97,1.09,and 0.90,respectively.4.C18H17NO6 nano-liposomes inhibited the proliferation of colon cancer cells in nude mice,and there was a significant time-effect relationship.On the 4th,8th,12th,16th and 20th day of the experiment,the tumor volume of the three groups was relative.The proliferation rate of ml experimental group was 91.72%,81.54%,78.73%,79.98%,and 66.57%,respectively.The m2 experimental group was 80.15%,69.52%,77.40%,70.05%,and 56.49%,respectively.The m3 experimental group was 76.65%,71.01%,70.94%,55.12%,and 50.98%,respectively.The relative proliferation rate of tumor volume decreased significantly with the increase of the number of administrations,which was the same as the tendency of C18H17NO6 nanoliposomes to inhibit colon cancer cells in vitro.And the higher the concentration of administration,the lower the tumor volume relative proliferation rate,and there is a dose-effect relationship.During the initial general toxicity experiment,with the increase of the number of doses,the body weight of the nude mice in the experimental group was similar to that of the negative control group,and the average body weight decreased slightly.On the contrary,the weight loss of the nude mice in the DDP positive control group was larger.This is related to the toxic side effects of the broad-spectrum anticancer drug cisplatin.At the end of the experiment,the nude mice were dissected and found to have certain damage to the liver in the experimental group,with little or no damage to other organs.5.By flow cytometry,it was found that C18H17NO6 nano-liposomes can promote the apoptosis of cancer cells,which can inhibit the proliferation and growth of cancer cells.Conclusion(s):1.The compound CxsH!7NO6 can be successfully prepared into nano-liposomes.The amount of soybean phospholipids and cholesterol is 5:1,the drug loading is 1.26 mg/mL,and the average particle size is less than 220 nm,which can be used for in vivo injection.The amount of soybean phospholipids and cholesterol is a factor that mainly affects the drug loading of nanoliposomes.2.Nano-liposomes have anti-tumor effect in vitro,and have a dose-effect relationship in vitro.With the increase of the concentration of the sample,the inhibition rate of the nano-liposomes is higher.The human colon cancer cell line(HCT-116)is the most sensitive cell line3.The combination of C18H17NO6nanoliposomes and cisplatin on the anti-tumor cells of XWLC-05 and MCF-7 showed additive effect.4.C18H17NO6 nano-liposomes in nude mice experiments have significant anti-tumor effect and dose-effect and time-effect relationship5.C18H17NO6 nanoliposomes can induce apoptosis of human colon cancer cell line(HCT-116). |
PDF Full Text Request