具有抗凝血活性的聚氨基酸的合成及性能研究
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摘要
通过亲核试剂三乙烯四胺(TETA)引发L-酪氨酸和L-谷氨酸的α-氨基酸N-环内酸酐(NCAs)的活性可控开环聚合(ROP),并进行氯磺酸酯化修饰,制备了一种具有抗凝血活性的磺酸酯化聚(L-酪氨酸-co-L-谷氨酸)(PTG-SO_3),并对PTG-SO_3的结构、酶促降解性能、细胞毒性以及血液相容性等进行了测试.结果表明,合成产物为PTG-SO_3,具有良好的生物降解性(16 d后可降解约65%)、低细胞毒性(培育72 h后,细胞活性约为70%)、良好的抗凝血性能和血液相容性(活化的部分凝血活酶时间延长约37 s).该聚合物有望应用于生物医学领域.
Using the nucleophilic reagent triethylenetetramine(TETA) as initiator, L-tyrosine and L-glutamic acid alpha amino acid N-carboxylic acid anhydride(NCAs) were actively controlled to occur the ring opening polymerization(ROP).Then,esterification modification of chlorosulfonic acid was prepared,a sulfonic acid esterification poly(L-tyrosine-co-L-glutamic acid)(PTG-SO_3) with anticoagulant activity had been prepared.The structure of PTG-SO_3,enzymatic degradation performance,cytotoxicity and blood compatibility were tested.The results showed that the product is PTG-SO_3 which has good biodegradability(about 65% biodegradable 16 d later),low cytotoxicity(after 72 h of culture,the cell activity is about 70%),favourable anticoagulant energy and blood compatibility(activated partial thrombin time can be extended by about 37 s).The polymer is expected to be used in biomedical applications.
Using the nucleophilic reagent triethylenetetramine(TETA) as initiator, L-tyrosine and L-glutamic acid alpha amino acid N-carboxylic acid anhydride(NCAs) were actively controlled to occur the ring opening polymerization(ROP).Then,esterification modification of chlorosulfonic acid was prepared,a sulfonic acid esterification poly(L-tyrosine-co-L-glutamic acid)(PTG-SO_3) with anticoagulant activity had been prepared.The structure of PTG-SO_3,enzymatic degradation performance,cytotoxicity and blood compatibility were tested.The results showed that the product is PTG-SO_3 which has good biodegradability(about 65% biodegradable 16 d later),low cytotoxicity(after 72 h of culture,the cell activity is about 70%),favourable anticoagulant energy and blood compatibility(activated partial thrombin time can be extended by about 37 s).The polymer is expected to be used in biomedical applications.
引文
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[2] QUAN M L,PINTO D J P,SMALLHEER J M,et al.Factor Ⅺa inhibitors as new anticoagulants[J].Journal of Medicinal Chemistry,2018,61(17):7425.
[3] CUI C,YANG Z,HU X,et al.Organic semiconducting nanoparticles as efficient photoacoustic agents for lightening early thrombus and monitoring thrombolysis in living mice[J].ACS Nano,2017,11(3):3298.
[4] 刘晋仙,李玮涛,张在忠,等.低分子肝素药理学机制及适应证研究进展[J].药学研究,2015,34(7):420.
[5] VALIMAKI S,KHAKALO A,ORA A,et al.Effect of PEG-PDMAEMA block copolymer architecture on polyelectrolyte complex formation with heparin[J].Biomacromolecules,2016,17(9):2891.
[6] PALUCK S J,NGUYEN T H,LEE J P,et al.A heparin-mimicking block copolymer both stabilizes and increases the activity of fibroblast growth factor 2(FGF2)[J].Biomacromolecules,2016,17(10):3386.
[7] NIE C X,MA L,CHENG C,et al.Nanofibrous heparin and heparin-mimicking multilayers as highly effective endothelialization and antithrombogenic coatings[J].Biomacromolecules,2015,16(3):992.
[8] LEI J H,YUAN Y Q,LYU Z L,et al.Deciphering the role of sulfonated unit in heparin-mimicking polymer to promote neural differentiation of embryonic stem cells[J].ACS Applied Materials Interfaces,2017,9(34):28209.
[9] MEAD G,HILEY M,NG T,et al.Directed polyvalent display of sulfated ligands on virus nanoparticles elicits heparin-like anticoagulant activity[J].Bioconjugate Chemistry,2014,25(8):1444.
[10] GU Y Y,WU X R,LIU H,et al.Photoswitchableheparinase Ⅲ for enzymatic preparation of low molecular weight heparin[J].Organic Letters,2018,20(1):48.
[11] ROBERTS J J,NAUDIYAL P,JUGE L,et al.Tailoring stimuli responsiveness using dynamic covalent cross-linking of poly(vinyl alcohol)-heparin hydrogels for controlled cell and growth factor delivery[J].ACS Biomaterials Science & Engineering,2015,1(12):1267.
[12] 王莺.肝素衍生物的抗凝血活性评价[J].中国医药指南,2013,11(17):81.
[13] PALUCK S J,NGUYEN T H,MAYNARD H D.Heparin-mimicking polymers:synthesis and biological applications[J].Biomacromolecules,2016,17(11):3417.
[14] 巫小莉,张玲,肖鸿岸,等.肝素体外诱导血小板聚集的相关性研究[J].实验与检验医学,2018,36(3):337.
[15] DENG J,LIU X Y,MA L,et al.Heparin-mimicking multilayer coating on polymeric membrane via LbL assembly of cyclodextrin-based supramolecules[J].ACS Applied Materials Interfaces,2014,6(23):21603.
[16] 文志红,邬素华,陈维涛.医用肝素化抗凝血高分子材料的研究进展[J].塑料,2005,34(2):26.
[17] 汤继辉,周建平,陈飞虎.聚氨基酸作为药物载体的研究进展[J].中国药科大学学报,2011,42(3):284.
[18] 徐虹,冯小海,徐得磊,等.聚氨基酸功能高分子的发展状况与应用前景[J].生物产业技术,2017(6):92.
[19] DENG C,WU J T,CHENG R,et al.Functional polypeptide and hybrid materials:precision synthesis via α-amino acid N-carboxyanhydride polymerization and emerging biomedical applications[J].Progress in Polymer Science,2014,39(2):330.
[20] ZHAO W,GNANOU Y,HADJICHRISTIDIS N.Fast and living ring-opening polymerization of alpha-amino acid N-carboxyanhydrides triggered by an“alliance”of primary and secondary amines at room temperature[J].Biomacromolecules,2015,16(4):1352.
[21] WANG L L,WU Y X,XU R W,et al.Synthesis and characterization of poly(L-glutamic acid-co-L-aspartic acid)[J].Polymer Science,2008,26:381.
[22] OHKAWA K,NAGAI T,NISHIDA A,et al.Purification of DOPA-containing foot proteins from green mussel,pernaviridis,and adhesive properties of synthetic model copolypeptides[J].Journal of Adhesion,2009,85:770.
[23] DATTA P P,KIESEWETTER M K.Controlled organocatalytic ring-opening polymerization of epsilon-thionocaprolactone[J].Macromolecules,2016,49:774.
[24] BHASKAR N,PADMAVATHY N,JAIN S,et al.Modulated in vitro biocompatibility of a unique cross-linked salicylic acid-poly(epsilon-caprolactone)-based biodegradable polymer[J].ACS Applied Materials Interfaces,2016,8:29721.
[25] CHOI Y C,CHOI J S,CHOI Y J,et al.Human gelatin tissue-adhesive hydrogels prepared by enzyme-mediated biosynthesis of DOPA and Fe3+ ion crosslinking[J].Journal of Materials Chemistry B,2014,2:201.
[26] SONG X,WANG K,TANG C Q,et al.Design of carrageenan-based heparin-mimetic gel beads as self-anticoagulant hemoperfusion adsorbents[J].Biomacromolecules,2018,19(6):1966.
[27] CUI Y Y,ZHOU F,BAI L H,et al.SEMA4D-heparin complexes immobilized on Titanium surfaces have anticoagulant,cell-migration-promoting,and immunoregulatory effects[J].ACS Biomaterials Science &Engineering,2018,4:1598.
[28] GAU A L,LIU F,XUE L X.Preparation and evaluation of heparin-immobilized poly(lactic acid)(PLA) membrane for hemodialysis[J].Journal of Membrane Science,2014,452:390.
[29] KUMAR V A,WICKREMASINGHE N C,SHI S,et al.Nanofibrous snake venom hemostat[J].ACS Biomaterials Science &Engineering,2015,1(12):1300.
[30] PAN Y,REN X T,WANG S,et al.Annexin V-conjugated mixed micelles as a potential drug delivery system for targeted thrombolysis[J].Biomacromolecules,2017,18(3):865.
[31] LU D D,ZHANG Y Y,LI Y F,et al.Preparation and properties of reversible hydrogels based on triblock poly(amino acid)s with tunable pH-responsivity across a broad range[J].Journal of Polymer Science Part A,2017,55(2):207.