Study On The Glycosyl Galactose Acyclovir Triparted Pre-drug Nanoparticle Delivery System

Objective: To prepare acyclovir triparted pre-drug and nanoparticle,the surface was modified by galactose then could overcome the limited access of drug to liver cells and increased liver target. acyclovir succinimide active ester was prepared by N-hydroxysuccinimide and SACV activated with DCC,BSA-ACV was prepared by BSA and acyclovir succinimide active ester, it's stability in different pH buffer solution was evaluated as well;Then make BSA-ACV into nanoparticle using desolvation preocess. Toxicity to HepG-2 cells and liver target in vivo were studied.Methods: The orthogonal design was applied to the optimization of the synthesis.The effects such as the rate of NHS and SACV, the rate of DCC and SACV, the reaction time,temperature and activation rate were investigated.The activation rate was 79.1% under the optimal condition,The synthesis conditions of BSA-ACV including BSA concentrantion, pH,reaction time and mole ratio were optimized in order to reach high couple rate,The concentrations of ACV in different pH buffer solution were detemined by HPLC,and then indirect calculated the concentrations of BSA-ACV,from which could deduce the functions.Nanoparticle was prepared by desolvation method ,the influence elements including BSA-ACV concentration,pH, alcohol consumption and the amount of curing agent were evaluated to receive the best properties of the morphology,size distribution,The optimal preparation elements were confirmed and coupled with glycosyl on nanoparticle surface.The products were analyzed and characterized by Spectrophotometer,FTIR,TEM,LPA(laser particle analyzer).The cytotoxicity of GANP on human liver cancer cells HepG-2 compared with ANP and ACV solution group was studied using MTT methord,The liver target of GANP and ANP were investigated after intraperitoneal injection in mice,Compared with ACV solution injection.Result: The activated SACV analyzed by FTIR had the characteristic peak of NHS, so it showed the product was acyclovir succinimide active ester,and activation rate was 79.1%, BSA-ACV analyzed by Spectrophotometer,FTIR,and DTA(Microcumputer Differential Thermal Analyzers) indicated that BSA connected some numbers of ACV ,and then couple rate of BSA-ACV measured by ninhydrin coloration method reached 24.1%.The study of BSA-ACV stability suggested that it display pseudo-first-order kinetics in buffer solutions,and It degrated faster when PH increased, and it's half-life was 58.73h when pH was 2.0,but 5.41h when 9.7,After anlyzed by TEM and LPA,It indicated that nanoparticle possess a smooth and round appearance and average diameter was 187nm,drug loading was 4.2237%,After release study in vitro showed that 56.19% of ACV could be released from GANP over a period of 48h.The GANP group had most apparent high rate of inhibition reached 96.5%,and IC50 was only 0.954 mg/mL,far less than the IC50 of acyclovir solution which was 8.853mg/mL.After intraperitoneal injection,maximally about 69.6% of the injucted dose was taken up by the liver,The AUC in liver was about 19.25 times of ANP group,34.05 Times of ACV solution group.Conclution: The results of the study indicated that the preparation of acyclovir for the triparted pre-drug and then prepared for nanoparticle whose surface coupled with glycosyl as a drug delivery systems(TDDS),Which could enhance the uptake of drugs into liver,was an effective strategy and a new approach for the treatment of hepatitisB.