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Preparation And Characteristics Of Novel Target Multi-modified Albumin Drug Delivery System By Coaxial-electrospray Technology

Posted on:2020-03-17Degree:MasterType:Thesis
Country:ChinaCandidate:S Z LuFull Text:PDF
GTID:2381330596991841Subject:Pharmaceutical Engineering
Abstract/Summary:PDF Full Text Request
The combined drug delivery strategy for traditional Chinese medicine bufalin(with excellent anti-tumor effect)and nintedanib(new multi-target tyrosine kinase inhibitors)was designed to improve the achieve synergistic anti-tumor effect anti-cancer.The novel delivery platform was constructed by coaxial electrospray technology with taking multi-modified albumin(which has targeting and synergistic tumor inhibition effects)as a carrier material to increase drug solubility,improve targeted delivery efficiency and reduce toxic and side effects.The application of the compound administration strategy and the novel technology could exert the anti-tumor effect of the drug,reduce the toxic and side effects,which would be a potential platform for the anti-tumor treatment of the traditional Chinese medicine monomer.This paper is divided into five sections as shown in the following.Part one:ReviewsIn this part,firstly,the research progress of bufalin(BF)and nintedanib(NDNB),and combined administration strategies were introduced.Secondly,the application of albumin carrier in drug delivery system is briefly described.The characteristics of biguanide group and ursodeoxycholic acid group(UA)in anti-tumor are analyzed.Finally,the characteristics and application of coaxial electrostatic spray technology,a new micro-nano particle preparation technology were introduced.Part two:Preformulation studiesIn this part,high performance liquid chromatography(HPLC)was established to determine the drug concentrations of BF and NDNB with good specificity,and the detection conformed the linear requirements in the concentration range of 1-25μg/mL.The quantitative limit and detection limit of BF was 75 ng/mL and 25 ng/ml,and the quantitative limit and detection limit of NDNB was 110 ng/mL and 40 ng/mL respectively.The other methodologies of the method were all complyed with the measurement requirements.The equilibrium solubility of BF in pH7.4 PBS and pH7.4 PBS(0.1%Tween80)solutions were 42.03±1.34μg/mL and 876.77±2.71μg/mL.The equilibrium solubility of NDNB in pH7.4 PBS and pH7.4 PBS(0.1%Tween80)solutions were 4.89±0.88μg/mL and 95.03±1.06μg/mL.The stability of the drug in different concentrations(1,10 and 25μg/mL),several mediums(ethanol,pH7.4 PBS,pH7.4 PBS(0.1%Tween 80)and pH7.4 PBS(0.1%Tween 80,0.5%serum))for a period of time all coincide with experimental demands.Part III:Preparation and Characterization of Bis-guanidino-ursodeoxycholic Acid Modified Bovine Serum AlbuminIn this part,firstly,The para-biguanidinyl benzoyl acid(DP)was synthesized and its structure was successfully characterized.After that,modified proteins UA-BSA,DP-BSA and DP-UA-BSA were prepared by Carbodiimide method.The fiuorescence spectrum and ultraviolet spectrum were used to obsereved the spectrum change of the modified albumin.The modification degree of UA in UA-BSA was determined to be 10.68 and 8.56 by fluorescent amine method and mass spectrometry respectively,and the two results were coincident,which indicated the successful modification of small molecules on albumin.In addition,the modification degree of DP in DP-BSA and DP-UA-BSA was determined to be 38.15 and 29.15by mass spectrometry,further proving the successful preparation of modified protein.Finally,the safety of the modified proteins was evaluated by MTT method,which displayed good biocompatibility in the concentration range of 50-500μg/mL.Part IV:Preparation of drug-loaded albumin microparticles by coaxial electrospray method and its in vitro propertiesIn this part,the albumin microspheres were successfully prepared by uniaxial electrospray technology,and the particle size was uniform,ranging from 300 nm to 900 nm.The drug-loaded albumin microparticles were prepared by coaxial electrospray technology.The single factors and response surface center composite design were used to optimize formulation of drug-loaded albumin microparticles.The characterization results showed that the particle size of the drug-loaded albumin microparticles was 879 nm±56 nm and the potential was-5.86±1.12 mV.The encapsu-lation efficiency of BF and NDNB were 78.0%and 76.2%and the drug loading of BF and NDNB were 1.57%and 2.20%.The results of SEM,TEM and LSCM showed that the drug-loaded albumin microparticles had good Spherical morphology and good core-shell structure.The XRD pattern showed that BF and NDNB was present in an amorphous state in the carrier.The results of in vitro release showed that the release rate of BF and NDNB in drug-loaded albumin microparticles was slower than free drug,and the dissolution and release behavior of the drug were coincident before and after adding serum with good stability.Part V:Antitumor effect,targeting effect and mechanism of compound multi-modified albumin microparticles in vitroThe MTT results showed that the inhibition of proliferation rates of various microparticles on tumor cells(HepG2 cell)were as follows:compound multi-modified albumin microparticles>compound single modified albumin microparticles>compound unmodified albumin microparticles>single drug multi-modified albumin microparticles>free drug.The combination index CI of BF and NDNB in the drug delivery system was calculated by the formula to be less than 1,indicating that the two drugs exert synergistic effects on HepG2 cell inhibition.The trend of drug IC500 in each modified protein-loaded microparticles indicated that the microparticles prepared by the multi-modified protein material has a dual promotion effect on HepG2 cell inhibition.Cellular uptake experiments indicated that ursodeoxycholic acid small molecules has targeting effect on tumor cells,and multi-modified albumin microparticles showed good targeting to HepG2 cell.PartⅥ:In vivo pharmacokinetics,tissue distribution and pharmacodynamics studiesIn this part,the in vivo analytical methods of BF and NDNB was established.And,the methodological verification of the method was carried out,which proved that the method was accurate and reliable to meet the requirements of methodology.In vivo pharmacokinetic results showed that the elimination half-life of BF in compound multi-modified albumin microparticles(BF-NDNB-DP-UA-BSA NPs)increased from 0.526 h to 2.948 h,and the mean residence time(MRT)also changed from 1.224 h to 4.154 h.The elimination half-life of NDNB increased from1.733 h to 4.130 h,and the mean residence time(MRT)changed from 2.064 h to 4.384 h,which indicated that the compound multi-modified albumin microparticles had obvious sustained release effect,and the targeting of compound multi-modified albumin microparticles was evaluated by in vivo imaging techniques,and the distribution of microparticles in mice was examined by isolated tissue.The results of in vivo imaging and isolated tissue distribution showed that the compound multi-modified albumin microparticles group received a significant increase in tumor uptake and a good active targeting effect compared with the unmodified preparation group.Pharmacodynamic evaluation resultsshowed that the prepared BF-NDNB-DP-UA-BSA NPs had the strongest anti-tumor effect.The combination of BF and NDNB could improve the anti-tumor effect of the preparation.The mulit-modification carrier could exert a dual anti-tumor effect through the anti-tumor effect of DP and the active targeting of UA,and enhance the anti-tumor effect.The safety evaluation results showed that BF-NDNB-DP-UA-BSA NPs can reduce the cardiotoxicity of active pharmaceutical ingredient and have better safety.
Keywords/Search Tags:bufalin, nintedanib, coaxial electrospray technology, albumin, biguanidine, ursodeoxycholic acid
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