Preparation And Evaluation Of Human Serum Albumin Modified Irinotecan Hydrochloride PLGA Nanoparticles

In this paper,Human serum albumin(HSA)and Nanoparticles(NPs)were combined to prepare a new drug delivery carrier,HSA-modified PLGA nanocarboxer(HSA/PLGA NPs),which was used as the model drug,Irinotecan hydrochloride(CPT-11),an antitumor drug.Human serum albumin modified irinotecan hydrochloride PLGA nanoparticles(HSA/PLGA/CPT-11 NPs)were prepared to enhance the drug targeting,reduce the serious adverse reactions of the existing marketed preparations of irinotecan hydrochloride in clinical application,so that irinotecan hydrochloride can safely and effectively play its broad spectrum anti-tumor effect.This paper mainly consists of four parts: the pre-prescription study of HSA/PLGA/CPT-11 NPs,the preparation and optimization of HSA/PLGA/CPT-11 NPs,the preparation and evaluation of HSA/PLGA/CPT-11 NPs,and the anti-tumor activity study of HSA/PLGA/CPT-11 NPs in vitro.Part Ⅰ: Pre-prescription study of HSA/PLGA/CPT-11 NPs.HPLC method was established to determine the content of HSA/PLGA/CPT-11 NPs,and the methodology was verified.The verification results showed that the established method had good specificity,high accuracy,high precision and good reproducibility.At the same time,the method of encapsulation rate and drug loading of HSA/PLGA/CPT-11 NPs was established.Part Ⅱ: Preparation and optimization of HSA/PLGA/CPT-11 NPs.In this paper,emulsification-solvent evaporation method was used to prepare carrier materials with HSA and PLGA.Single factor investigation and response surface methodology were used to optimize the formulation process.The optimal formulation was determined as HSA concentration 2.65%,PLGA dosage 86.16 mg,and chloroform: ethanol =4.25:1(v/v).Three batches of HSA/PLGA/CPT-11 NPs were prepared according to the optimized prescription,and the average particle size was(97.51±1.22)nm,the average PDI was(0.220±0.01),the average encapsulation rate was(67.03±0.71)%,the average drug loading was(1.02±0.01)%,the average potential was(-20.07±0.17)m V,and the average p H was(5.86±0.01).Part Ⅲ: Preparation and evaluation of freeze-dried HSA/PLGA/CPT-11 NPs.In order to improve the stability of HSA/PLGA/CPT-11 NPs preparation and prolong the storage time,HSA/PLGA/CPT-11 NPs colloid solution was prepared into HSA/PLGA/CPT-11 NPs freeze-dried product.Through the screening of the type and dosage of the lyophilized protectant,it was determined that 1% mannitol was the protective agent added in the lyophilized process of HSA/PLGA/CPT-11 NPs,and the retention rate of lyophilized agent was 96.14%.After lyophilized,the average particle size was(98.73±2.04)nm,the average PDI was(0.257±0.03),the average potential was(-18.30±0.14)m V,the average encapsulation rate was(64.61±0.29)%,and the average drug loading was(1.00±0.01)%.The morphology of the particles was spherical and the outer layer of HSA was obvious under TEM.The results of FTIR characterization of HSA/PLGA/CPT-11 NPs showed that CPT-11 was not simply physically mixed with the carrier,but formed a new phase after being embedded by the carrier.The release of CPT-11 and HSA/PLGA/CPT-11 NPs in the release medium of p H7.4 and 5.0 PBS buffer was investigated by dialysis bag method.The results showed that HSA/PLGA/CPT-11 NPs had a certain slow-release property.The preliminary stability of HSA/PLGA/CPT-11 NPs was investigated.The results showed that the freeze-dried HSA/PLGA/CPT-11 NPs needed to be stored at low temperature and away from light.The physical and chemical properties of the freeze-dried HSA/PLGA/CPT-11 NPs did not change significantly when it was stored at 4℃ and 25℃ for 2 months.Part Ⅳ: Anti-tumor activity of HSA/PLGA/CPT-11 NPs freeze-dried product in vitro.The cytotoxicity of HSA/PLGA NPs on human colon cancer HCT-116 cells and the antitumor activity of HSA/PLGA/CPT-11 NPs on HCT-116 cells were investigated by CCK-8 assay.The results showed that the cytotoxicity of HSA/PLGA NPs on HCT-116 cells was small,but had no effect on the antitumor activity of HSA/PLGA/CPT-11 NPs on HCT-116 cells.The results of anti-tumor activity showed that HSA/PLGA/CPT-11 NPs inhibited HCT-116 cells more than CPT-11 cells in a concentration-dependent manner.In conclusion,the freeze-dried HSA/PLGA/CPT-11 NPs prepared in this paper is of good quality and has obvious anti-tumor effect in vitro,which provides ideas for the development and clinical application of new preparations of CPT-11.