Preparation Of Cabazitaxel-Loaded Bovine Serum Albumin Nanoparticles And Anti-Prostate Cancer Effect

Cabazitaxel is a new type of paclitaxel antineoplastic drug approved by FDA for the treatment of prostate cancer,especially for patients with advanced prostate cancer who are ineffective or even aggravated by paclitaxel.However,due to the addition of Tween-80,some patients are prone to serious allergic reactions in the prescription of cabazitaxel injections,and the toxicity,adverse reactions and poor tumor selectivity of cabazitaxel itself make the clinical application greatly restricted.Therefore,the preparation of cabazitaxelloaded bovine serum albumin-Gd nanoparticles(CBZ-BSA-Gd-NPs)were developed in this study to improve the solubility,bioavailability and safety of clinical treatment of cabazitaxel,enhance the clinical anti-tumor effects,achieve the treatment effects of high efficiency and low toxicity,and provide a new choice for the further clinical application of cabazitaxel.In the first chapter,a method in vitro for the determination of cabazitaxel was established to determine the encapsulation rate and leakage rate of nanoparticles.Then,cabazitaxel-loaded bovine albumin-Gd nanoparticles were prepared by biomineralization method.The properties of encapsulation efficiency and drug loading were characterized,and the formulation of nanoparticles was optimized by orthogonal design.In the second part,the in vitro biosafety and efficacy of CBZ-BSA-Gd-NPs were evaluated.The safety of nanoparticles was determined by in vitro hemolysis test,and the efficacy was determined by cell uptake test and cytotoxicity test.In the third chapter,an UHPLC/MS/MS method for the determination of the content of cabazitaxel in vivo was constructed,and the methodology was investigated.Then,the pharmacokinetic behavior in vivo of cabazitaxel in CBZ-BSAGd-NPs were studied by measuring the concentration of the drug in rat plasma after administration.And the subcutaneous prostate cancer model of nude mice was established,the tissue distribution of cabazitaxel were also demonstrated by determining the content of cabazitaxel in different tissues by UHPLC-MS/MS,whose results indicating that concentration of cabazitaxel in cancer tissue were higher and that of other organ tissues were lower.In the fourth part,the subcutaneous prostate cancer model of nude mice was established,and the changes of body weight and tumor volume with time were recorded after the injection of CBZ-BSA-Gd-NPs and cabazitaxel Tween-80 injection,respectively.The therapeutic effect and safety of the constructed cabazitaxel bovine serum albumin nanoparticles on prostate cancer were evaluated.The results showed that HPLC method for the determination of cabazitaxel was specific,linear,limit of detection,limit of quantification,recovery,precision and repeatability were all in line with the requirements.It can be used for the determination of cabazitaxel in CBZBSA-Gd-NPs.And CBZ-BSA-Gd-NPs were prepared of by biomineralization,and its physical and chemical properties were studied.The results showed that the encapsulation efficiency of the nanoparticles was 63.04%,the drug loading was 10.51%,the average particle size was(166.1 ± 4.7)nm,the PDI of the nanoparticles was 0.256,and the zeta potential was(-18.14 ± 1.16)mv.The results of particle size change experiment and in vitro leakage experiment within 24 hours showed that the nanoparticles are stable in nature.The relaxation evaluation results of nanoparticles showed that CBZ-BSA-Gd-NPs prepared in this study have high imaging efficiency and are expected to be used for tumor diagnosis.The results of in vitro hemolysis experiment of CBZ-BSA-Gd-NPs showed that compared with carbataxel-Tween-80 solution,the hemolysis effect of CBZ-BSA-Gd-NPs in vitro was significantly reduced;the results of tumor cell inhibition showed that the tumor inhibition effect of CBZ-BSA-Gd-NPs was not decreased,and it had more cell uptake ability,so it can play a better role in anti-tumor in addition to avoiding allergic reaction.The results of pharmacokinetic experiments in rats showed that after loading albumin nanoparticles,compared with carbataxel-Tween-80 injection,the retention time of CBZ-BSA-Gd-NPs in vivo was longer,the removal speed was slower,the drug concentration in blood could be kept higher,the area under the drug time curve was increased,which was conducive to the development of drug efficacy.The experimental results of tumor inhibition showed that the content of cabazitaxel nanoparticles in heart,liver,spleen and kidney was lower than that of cabazitaxel-Tween-80 injection,and the content in tumor tissue was higher,which was mainly due to the high permeability and retention effect of nanoparticles.To sum up,the preparation method of CBZ-BSA-Gd-NPs involved in this study is simple,highly repetitive,without the addition of organic solvents,safe and environmental protection.The prepared CBZ-BSA-Gd-NPs can significantly prolong the in vivo action time of cabazitaxel drugs,significantly improve the hepatorenal toxicity caused by the addition of Tween 80 and other solvents,and also reduce hemolysis.The results of pharmacological evaluation showed that after the administration of CBZ-BSA-Gd-NPs in prostate cancer nude mice,the tumor inhibition effects were more significant compared with that of cabazitaxel-Tween 80 injection.Therefore,it can achieve the treatment effect of high efficiency and low toxicity.