| Background:Curcumin has a variety of pharmacological activities,especially its significant anti-tumor activity,but due to its poor solubility,fast metabolism,and low bioavailability,its application is limited.How to improve the bioavailability of curcumin is an important practical problem faced by the development and utilization of curcumin.At present,there are two main means to solve this problem,one is to modify the structure of curcumin,and the other is to prepare a suitable nanoparticle.Through preliminary experiments,a series of curcumin derivatives were evaluated for pharmacokinetics and efficacy to select drugs with strong activity and long half-life.Finally,we selected curcumin derivative C210 for the design of nanoparticle in order to further enhance its anti-tumor effect in vivo.Objective:To construct C210 nanocrystals delivery system,study its anti-tumor activity and pharmacokinetic.Methods:1.The best preparation method of C210 nanocrystals and RBC membrane-coated C210 nanocrystals were investigated by single factor method and its pharmaceutical properties were characterized.Firstly,its Size and Zeta potential were studied,and its stability and biocompatibility were investigated.2.The MTT assay was used for detection the activity of proliferation inhibition of C210 nanocrystals on Hep G2 liver cancer cell.3.The plasma concentrations at various time points for the original drug and C210 nanoparticles were determined to draw curve of drug-time and calculate the pharmacokinetic parameters by HPLC.Results:1.The optimum process were investigated by single factor method.The film dispersion method was the best preparation method,F127 was the best stabilizer,0.9%Na Cl solution was the best hydration solution,and the best ratio of drug to stabilizer was 1:4.The characterizations of C210 nanocrystals were determined by transmission electron microscopy and laser particle size analyzer.The particle size of C210 nanocrystals was 52.59 ± 0.33 nm and the Zeta potential was-2.5 ± 0.4 m V.We found that it was difficult to keep stability for a long time.The hemolysis experiment proved that the nanacrystals concentration of 0-100 μg/ml had a hemolysis rate of less than 3%to the blood of mice.2.The optimum process were investigated by single factor method.Ultrasonic breaking method was the best preparation method of RBC membrane vesicles and the best ratio of red blood cell membrane to drug was 2:1.The characterizations of RBC membrane-coated C210 nanocrystals were determined by transmission electron microscopy and laser particle size analyzer.The particle size was 181.85±0.33 nm and the Zeta potential was-20.4±0.7 m V.RBC membrane-coated C210 nanocrystals could keep stability within 48 h.The hemolysis experiment proved that the nanacrystals concentration of 0-100 μg/ml had a hemolysis rate of less than 3% to the blood of mice.It was proved that RBC membrane coated on the surface of C210 NC still retained the original membrane surface protein by SDS-PAGE gel electrophoresis.3.In the MTT experiment,C210 and two kinds of nanoparticles showed a strong proliferation inhibitory effect after 48 h on Hep G2 cells.Through trypan blue dye exclusion test,it was found that the two kinds of nanoparticles showed stronger proliferation inhibition than the original drug at two concentrations of 0.5 and 1μg/ml.4.The C210 concentrations in rat plasma were detected by HPLC,and HPLC method showed that it was simple,rapid,specific,accurate and reliable,and conform to the analytic requirements of plasma samples.The results were calculated by software.The elimination phase half-lives of C210、C210NC and RBC-C210 NC were 42.28 ±8.04、24.70 ± 17.12 and 73.75 ± 18.63 min,MRT were 58.78 ± 3.50、20.98 ± 26.81 and95.78 ± 34.76 min.This indicated that RBC-C210 NC had a longer half-life and residence time compared to the original drug.Conclusions:1.We have successfully produced C210 nanocrystals,and find the best preparation method,preparation prescription and preparation process.The nanocrystals have good biocompatibility,but its stability is poor.2.We have successfully prepared RBC membrane-coated C210 nanocrystals,and find the best preparation method and preparation prescription.The nanoparticles have good stability and biocompatibility.3.Both of the C210 nanoparticles show a better killing effect for tumor cells than the original drug in vitro anti-tumor experiments.4.Pharmacokinetic experiment results show that RBC-C210 NC has a longer half-life in vivo than the original drug,which is beneficial to improve the therapeutic effect of the drug in vivo. |
