Study Of Preparation Andpreliminary Evaluation Of Albuminnanoparticle Loaded With Indirubin

The indole alkaloids of indirubinia indirubinum(IND),mainly derived from traditional Chinese medicine Qingdai and Daqingye,are used in the treatment of chronic myeloid leukemia(Chronic Myelogenous Leukemia,CML).Modern studies show that they have a wide range of pharmacological effects,but indirubin is a hydrophobic drug,its low solubility and low bioavailability limit its clinical use.In this study,a new preparation,albumin nanoparticle loaded with indirubin(IND-HSA-NPs)was developed by using human serum albumin(HSA)serum albumin as carrier.The optimum formulation and preparation technology were selected by single factor investigation and orthogonal experiment.And the pharmaceutics evaluation,pharmacokinetics,tissue distribution and preliminary pharmacodynamic studies were carried out.In this paper,the preparation of IND-HSA-NPs by emulsification-chemical crosslinking method was established,and the effects of organic phase,ratio of organic phase to water phase,ultrasonic time and ultrasonic power on IND-HSA-NPs were investigated by single factor experiment.The optimum formulation and preparation technology of IND-HSA-NPs were obtained by orthogonal experiment.Through the Malvin laser particle size analyzer measured the average particle size of IND-HSA-NPs was 334.5 nm,PDI was 0.06,the average Zeta potential was-23.4m V.IND-HSA-NPs were observed by scanning electron microscope,the results showed that the nanoparticles were spherical or spherical.The encapsulation efficiency of IND-HSA-NPs was 80.15± 2.47% and the drug loading was 2.13±0.42%.by ultra high speed freezing centrifugation.In addition,a HPLC method for the determination of indirubin was established.The specificity and precision of the method met the requirements of the methodology.The results of in vitro drug release test showed that albumin encapsulation could significantly delay the release of indirubin for 48 h and the cumulative release rate was about60%.The pharmacokinetic study showed that the maximum plasma concentration(Cmax)in IND-HSA-NPs group was lower than that in the indirubin raw drug group(free IND)under the same dosage of indirubin,and the area under the drug time curve(AUC0-∞),the half-life(T1/2),and the mean residence time(MRT)were 1.44~2.68,1.22~2.67,1.10~1.33 times of the free IND group.The plasma clearance rate was lower than that of free IND group.When indirubin was made into albumin nanoparticles,the plasma concentration could be maintained for a long time,showing a certain sustained release.The tissue distribution in mice showed that the concentration of heart,liver,spleen,lung and kidney in IND-HSA-NPs group was higher than that in the free IND group.The content of IND-HSA-NPs in each tissue was significantly higher in 0.25 h group than that in free IND group,and the drug concentration in each tissue decreased with time.The drug concentration of IND-HSA-NPs group decreased first quickly and then slowly,and even more slowly after 12 hours.It shows the slow release of IND-HSA-NPs.Preliminary pharmacodynamics studies showed that different doses of preparation had a certain inhibitory effect on K562 solid tumor.The effect of IND-HSA-NPs group was better than free IND group.By analyzing the tumor inhibition rate and tumor volume,we found that the best inhibition rate in the IND-HSA-NPs(10mg/kg)group was 67.5%.The effect of IND-HSA-NPs(5mg/kg)was similar to free IND(10mg/kg),and the inhibitory rate was47.5%,67.5%.The pictures of tumor volume more intuitively reflect the advantage of IND-HSA-NPs group over free IND group in solid tumor treatment.