Haluronic Acid Modified Cationic Nanoparticles To Overcome Enzyme CYP1B1 Medicated Multidrug Resistance

Objection: HA modified cationic Poly Lactic-co-Glycolic Acid(PLGA)nanoparticles were developed for co-delivery of docetaxel(DTX)and the CYP1B1 enzyme inhibitor ?-naphthoflavone(ANF).In order to targeting DTX to tumor site and achieving an effective antitumor effect.The physicochemical properties of the nanoparticles will be explored in vitro and in vivo.Methods: Establishing in vitro analysis method of DTX and ANF.DTX and ANF loaded PLGA nanoparticles were prepared by emulsion-evaporation method after formulation optimizing by one-way analysis.Morphology of HA/PEI-(DTX+ANF)-NPs was observed by transmission electron microscopy(TEM).Mean particle size and Zeta potential were measured by using Zetasizer.Encapsulation efficiency and drug loading of DTX and ANF were analyzed by HPLC.The vitro release was evaluated using dialysis bag technique in the medium at p H 7.4.To evaluate the vitro features of HA/PEI-(DTX+ANF)-NPs in cell level,the cell viability measured by MTT method,cell apoptosis measured by Annexin V-FITC apoptosis detection kit were investigated.Moreover,CLSM and flow cytometer were used to measure cell cellular uptake and intracellular distribution of NPs on MCF-7 cells and MCF-7/1B1 cells.And the pharmacokinetic characteristics of HA/PEI-(DTX+ANF)-NPs were studied in the SD rats.Results: Established in vitro HPLC analysis method of DTX and ANF successfully.The constructed HA/PEI-(DTX+ANF)-NPs exhibited typical spherical morphology with uniform size of(184.8±8.7)nm and zeta-potential of(-16.31±1.28)m V.The drug encapsulation efficiency and drug loading were(82.76±6.08)%,(90.67±8.22)% and(1.14±0.18)%,(1.56±0.20)% for DTX and ANF respectively.In vitro release showed a sustained release behavior.In vitro cell level experiments,the HA/PEI-(DTX+ANF)-NPs showed the most cytotoxicity among the experimental groups and promoted the apoptosis of MCF-7/1B1 cells remarkably.The CLSM and flow cytometer results demonstrated the HA/PEI-(DTX+ANF)-NPs could significantly enhance cellular uptake.The pharmacokinetic results indicated the HA/PEI-(DTX+ANF)-NPs could improve the bioavailability of free DTX.Conclusion: The HA/PEI-(DTX+ANF)-NPs could target to tumor site and released drug slowly,improve the bioavailability of the DTX and overcome multidrug resistance.