聚苹果酸合成新方法及其聚苹果酸苄基酯用作药物载体的研究
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摘要
目的:优化β-聚苹果酸(PMLA)的合成路线,制备部分氢化的聚苹果酸苄基酯(PMLABz),为构建载药纳米胶束提供两亲性聚合物载体。方法:分别以L-天冬氨酸和L-苹果酸为原料,合成单体β-苄氧羰基-β-丙内酯,再通过阴离子开环聚合制备PMLABz,最终氢化制备PMLA。X射线衍射仪、差示扫描量热仪等对材料的性质进行表征,L929成纤维细胞测定聚合物的细胞毒性,透析法制备部分氢化的聚苹果酸苄基酯空白胶束。结果:通过优化反应条件,以L-苹果酸为原料合成关键中间体β-苄氧羰基-β-丙内酯的终产率为31.5%,对比文献报道的L-天冬氨酸合成路线,产率提高近7倍。X-RD结果表明以L-苹果酸为原料制备的PMLABz有明显的衍射峰,结晶度高,熔点随之升高,MTT实验证实PMLABz无毒性。部分氢化的PMLABz可在水中自组装成一定粒径的胶束,氢化77%的PMLABz可得到粒径均匀、形态良好的纳米胶束。结论:分别以L-天冬氨酸和L-苹果酸为原料合成聚苹果酸,优化合成路线,提高终产率,得到了新晶型PMLABz,并通过部分氢化PMLABz制备两亲性聚合物进而得到纳米胶束,为后期纳米释药体系研究提供优良载体。
Objective: To optimize the synthesis route of PMLA and prepare the partially hydrogenated poly(β-malic acid benzyl ester) which is to provide the amphiphilic polymer carrier for the drug-loaded nanomicelles.Methods: Benzyl-β-malolactonate(MLABz)was prepared from L-aspartic acid and L-malic acid,respectively.The properties of the polymers were characterized by X-RD and DSC.The cytotoxicity of PMLABz was assessed by using L929 fibroblasts.The partial hydrogenation of PMLABz was prepared by controlling the hydrogenation time to obtain an amphiphilic block copolymer.Results: The yield of MLABz from L-malic acid was 31.5 % nearly seven times as much as the L-aspartic acid by optimizing the reaction conditions.The X-RD patterns revealed that the PM-LABz-2 had obvious diffraction peaks.The melting temperature was increased due to the high crystallinity.MTT test confirmed that the PMLABz was nontoxic.The partial hydrogenation of PMLABz could form the certain size of the nanomicelles by self-assembling in water.It was confirmed that a hydrogenation degree was 77 % of PMLABz which would obtain the well-formed nanomicelles.Conclusions: MLABz was prepared from L-aspartic acid and L-malic acid,respectively.The synthesis routes of PMLA from L-malic acid had the short route and high yield.Meanwhile,a novel crystalline PMLABz that polymerized by MLABz starting from L-malic acid was discovered.An amphiphilic block polymer was obtained by partial hydrogenolysis of PMLABz which would provide an excellent nanocarrier for drug delivery system.
Objective: To optimize the synthesis route of PMLA and prepare the partially hydrogenated poly(β-malic acid benzyl ester) which is to provide the amphiphilic polymer carrier for the drug-loaded nanomicelles.Methods: Benzyl-β-malolactonate(MLABz)was prepared from L-aspartic acid and L-malic acid,respectively.The properties of the polymers were characterized by X-RD and DSC.The cytotoxicity of PMLABz was assessed by using L929 fibroblasts.The partial hydrogenation of PMLABz was prepared by controlling the hydrogenation time to obtain an amphiphilic block copolymer.Results: The yield of MLABz from L-malic acid was 31.5 % nearly seven times as much as the L-aspartic acid by optimizing the reaction conditions.The X-RD patterns revealed that the PM-LABz-2 had obvious diffraction peaks.The melting temperature was increased due to the high crystallinity.MTT test confirmed that the PMLABz was nontoxic.The partial hydrogenation of PMLABz could form the certain size of the nanomicelles by self-assembling in water.It was confirmed that a hydrogenation degree was 77 % of PMLABz which would obtain the well-formed nanomicelles.Conclusions: MLABz was prepared from L-aspartic acid and L-malic acid,respectively.The synthesis routes of PMLA from L-malic acid had the short route and high yield.Meanwhile,a novel crystalline PMLABz that polymerized by MLABz starting from L-malic acid was discovered.An amphiphilic block polymer was obtained by partial hydrogenolysis of PMLABz which would provide an excellent nanocarrier for drug delivery system.
引文
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[2]Ding H,Gustavo H,JoséA.R,et al.Polymalic acid nanobioconjugate for simultaneous immunostimulation and inhibition of tumor growth in HER2/neu-positive breast cancer[J].J Controlled Release,2013,171(3):322-329
[3]He B,Wan YQ,Bei JZ.et al.Synthesis and cell affinity of functionalized poly(l-lactide-co-β-malic acid)with high molecular weight[J].Biomaterials,2004,25(22):5239-5247
[4]Huang ZW,Véronique L,Ghizlane C,et al.New functional degradable and bio-compatible nanoparticles based on poly(malic acid)derivatives for site-specific anti-cancer drug delivery[J].Int J Pharm,2012,423(1):84-92
[5]Loyer P,Cammas-Marion.Natural and synthetic poly(malic acid)-based derivates:a family of versatile biopolymers for the design of drug nanocarriers[J].J Drug Target,2014,22(7):556-575
[6]Pennapa M,Christopher D.S,Stephen W.P,et al.Poly(β-L-malic acid)production by diverse phylogenetic clades of Aureobasidium pullulans[J].J Ind Microbiol Biotechnol,2012,39(1):125-132
[7]Sandrine C,Isabelle R,Val6rie L,et al.Poly(β-malic acid):obtaining high molecular weights by improvement of the synthesis route[J].Polymer,1996,37(18):4215-4220
[8]Qiao YB,Duan X,Fan L,et al.Synthesis of controlled molecular weight poly(β-malic acid)and conjugation with HCPT as a polymeric drug carrier[J].J Polym Res,2014,21(4):397-405
[9]RuairíP.B,Andrew P.D.Synthesis,properties and biomedical applications of hydrolytically degradable materials based on aliphatic polyesters and polycarbonates[J].Biomater Sci,2016,5(1):9-21
[10]Taghavi S,Ramezani M,Alibolandi M,et al.Chitosan-modified PLGA nanoparticles tagged with 5TR1 aptamer for in vivo tumor-targeted drug delivery[J].Cancer Lett,2017,400:1-8
[11]Giarra S,Serri C,Russo L,et al.Spontaneous arrangement of a tumor targeting hyaluronic acid shell on irinotecan loaded PLGA nanoparticles[J].Carbohydr Polym,2016,140:400-407
[12]Jelonek K,Li SM,Kaczmarczyk B,et al.Multidrug PLA-PEG filomicelles for concurrent delivery of anticancer drugs-The influence of drug-drug and drug-polymer interactions on drug loading and release properties[J].Int J Pharm,2016,510(1):365-374
[13]Tan F,Mo XH,Zhao J,et al.A novel delivery vector for targeted delivery of the antiangiogenic drug paclitaxel to angiogenic blood vessels:TLTYTWS-conjugated PEG-PLA nanoparticles[J].J Nanopart Res,2017,19(2):1388-0764
[14]Ljubimova JY,Portilla-Arias J,Patil R,et al.Toxicity and efficacy evaluation of multiple targeted polymalic acid conjugates for triplenegative breast cancer treatment[J].J Drug Targets,2013,21:956-967
[15]Zhou Q,Hou YL,Zhang L,et al.Dual-p H Sensitive Charge-reversal Nanocomplex for Tumor-targeted Drug Delivery with Enhanced Anticancer Activity[J].Theranostics,2017,7(7):1806-1819
[16]Zhou Q,Yang TH,Qiao YB,et al.Preparation of poly(β-L-malic acid)-based charge-conversional nanoconjugates for tumor-specific uptake and cellular delivery[J].Int J Nanomedicine,2015,10:1941-1952
[17]Ding H,Portilla-Arias J,Patil R,et al.Distinct mechanisms of membrane permeation induced by two poly malic acid copolymers[J].Biomaterials,2013,34:217-225
[18]Nivishna V,Pascal L,Sandrine CM,et al.Poly(malic acid)bearing Doxorubicin and N-Acetyl Galactosamine as a site-specific prodrug for targeting hepatocellular carcinoma[J].J Biomater Sci Polym Ed,2017,28(10-12):1140-1157
[19]Caruelle,JP,Barritault D,Jeanbat-Mimaud V,et al.Bioactive functionalized polymer of malic acid for bone repair and muscle regeneration[J].J Biomater Sci Polym Ed,2000,11(9):979-991
[20]Patil R,Portilla-Arias J,Ding H,et al.Temozolomide delivery to tumor cells by a multifunctional nano vehicle based on poly(β-L-malic acid)[J].Pharm Res,2010,27(11):2317-2329
[21]Yang TH,Li W,Duan X.Preparation of Two Types of Polymeric Micelles Based on Poly(β-L-Malic Acid)for Antitumor Drug Delivery[J].PLo S ONE,2016,11(9):1-18
[22]Wang J,Zhang YN,Zhang M,et al.Preparation and p H controlled release of polyelectrolyte complex of poly(l-malic acid-co-d,l-lactic acid)and chitosan[J].Colloids Surf B:Biointerfaces,2014,115:275-279
[23]崔明凤,郭松岩,周青,等.新型生物材料聚(β-苹果酸-co-β-羟基丁酸酯)的合成和结构表征[J].材料导报,2015,2:224-228Cui Ming-feng,Guo Song-Yan,Zhou Qing,et al.Synthesis and Structural Characterization of New Biological Materials Poly(β-malicacid-co-β-butyrolactone)[J].Materials Review,2015,2:224-228
[24]He B,M.B.Chan-Park.Synthesis and Characterization of Functionalizable and Photopatternable Poly(ε-caprolactone-co RS-β-malic acid)[J].Macromolecules,2005,38(20):8227-8234