三肽纤维素酯的制备及应用研究

doi:10.3969/j.issn.0253-2417.2016.02.011

摘要/Abstract

摘要： 以微晶纤维素为原料,芴甲氧羰基-L-脯氨酸(FMOC-L-Pro-OH)、芴甲氧羰基-L-缬氨酰-L-苯丙氨酸(FMOC-L-Val-L-Phe-OH)和3,5-二硝基苯甲酸为衍生化试剂,经酰氯化、酯化、肽链的延长和氨基的修饰等反应,合成了末端氨基被修饰的三肽纤维素酯(TPC)。通过实验证实三肽若要与纤维素成酯,必须采用先酯化后肽链延长的方式,其中关键步骤为酯化反应,其反应条件为:纤维素在LiCl/DMAc溶液中的质量浓度为20 g/L,温度100 ℃,n(酰氯):n(纤维素中羟基总量)为3:1,反应时间20 h。再通过肽链延长、氨基修饰反应,最终可得到取代度为2.15的三肽纤维素酯。经过FT-IR、TG、XRD和元素分析等多种分析手段测试可知,3,5-二硝基苯甲酸对肽链末端进行了成功的修饰,该反应路径可行。酯化反应的产物根据沉淀析出液不同,内部氢键产生也不相同,可得到功能不同的三肽纤维素酯。TPC的应用实验结果表明,以甲醇为沉淀析出液的纤维素酯,是良好的肠溶包衣材料,具有遮光性强崩解迅速的特点,但其生理毒性有待进一步研究;以水为沉淀析出液的纤维素酯,被制成手性涂敷柱后,综合了刷型固定相与纤维素固定相的两大优势,拆分能力强大高效,耐化学性极佳,适用范围比以往涂敷柱更广泛,尤其适用于四氢呋喃流动相体系。

关键词: 微晶纤维素, 肽, 衍生物, 酯化, 合成

Abstract: Tripeptide-cellulose ester (TPC) was synthesized by sequent acyl chlorination, esterification, peptide elongation and amino modification using microcrystalline cellulose as raw material and N-(9-fluorenylmethoxycarbonyl)-L-proline(FMOC-L-Pro-OH), N-(9-fluorenylmethoxycarbonyl)-L-valinamide-L-phenylalanine(FMOC-L-Val-L-Phe-OH), 3,5-dinitrobenzoic acid as derivatization reagents. The experimental results showed that during the synthesis process, esterification, a key step, was performed firstly, and followed by the extension of peptide chain. The effects of various experimental factors on esterification and substitution degree (D_S) were investigated. The optimal condition was described as that the mass concentration of avicel in LiCl/DMAc 20 g/L, the molar ratio of —C(O)Cl to the hydroxyl content of cellulose 3:1, reaction temperature 100 ℃ and reaction time 20 h. By the further peptide elongation and amino modification, the tripeptide-cellulose ester with D_S 2.15 was synthesized. The structure and properties were then characterized by using Fourier transform infrared spectroscopy, elemental analysis, X-ray diffraction and thermogravimetric analysis. The results showed that the modification of peptide by 3,5-dinitrobenzoic acid was successful and the synthesis process was feasible. With the alteration of precipitants, the intramolecular hydrogen bond was different and the obtained derivatives had different characteristics and functionalities. The tripeptide-cellulose ester precipitated in methanol was good enteric-coating material with good shading effect and rapid disintegration, but the physiological toxicity needed further study. The tripeptide-cellulose ester (using water as the precipitant) could be coated onto silica gel to get a chiral stationary phase, which has a wider application than previous coated-cellulose chiral stationary phase. In addition, the stable chemical resistance of tripeptide-cellulose ester was particularly suitable for the mobile phase including tetrahydrofuran.

Key words: microcrystalline cellulose, peptide, derivatives, esterification, synthesis

中图分类号:

TQ35

李杨, 朱进科, 李连杰, 蒋登高. 三肽纤维素酯的制备及应用研究[J]. 林产化学与工业, 2016, 36(2): 71-78.

LI Yang, ZHU Jin-ke, LI Lian-jie, JIANG Deng-gao. Preparation and Application of Tripeptide-cellulose Ester[J]. Chemistry and Industry of Forest Products, 2016, 36(2): 71-78.

参考文献

[1] 毕殿洲.药剂学[M].4版.北京:人民卫生出版社,2001:57-60. BI Dian-zhou.Pharmacy[M].4th ed.Beijing:People's Medical Press,2001:57-60.
[2] 袁黎明.手性识别材料[M].北京:科学出版社,2010:220-221. YUAN Li-ming.Chiral Recognition Materials[M].Beijing:Science Press,2010:220-221.
[3] 武秀明,周成,王安明,等.化学-酶法合成多肽的研究进展[J].分子催化,2009,23(5):477-481. WU Xiu-ming,ZHOU Cheng,WANG An-ming,et al.Research progress of the synthesis of polypeptide in chemo enzymatic[J].Journal of Molecular Catalysis,2009,23(5):477-481.
[4] EDWARDS J,PREVOST N,SETHUMADHAVAN K,et al.Pepetide conjugated cellulose nanocrystals with sensitive human neutrophil elastase sensor activity[J].Cellulose,2013,20(3):1223-1235.
[5] DEVARAYAN K,HACHISU M,ARAKI J,et al.Synthesis of peptide-cellulose conjugate mediated by a soluble cellulose derivative having β-Ala esters(Ⅱ):Conjugates with O-phospho-L-serine-contatining peptides[J].Cellulose,2013,20(1):365-378.
[6] OHKAWA K,NISHIBAYASHI M,DEVARAYAN K,et al.Synthesis of peptide-cellulose conjugate mediated by a soluble cellulose derivative having β-Ala esters[J].International Journal of Biological Macromolecules,2013,53:150-159.
[7] OHKAWA K,HACHISU M,DEVARAYAN K,et al.Design and synthesis of peptide-cellulose conjugate molecules aspects from energy/steric profiles[J].Fibers and Polymers,2013,14(12):1970-1974.
[8] RAJESH K.生物药剂学在药物研发中的应用[M].宁保明,译.北京:北京大学医学出版社,2012:108-109. RAJESH K.Biopharmaceutics Applications in Drug Development[M].Translation by NING Bao-ming.Beijing:Peking University Medical Press,2012:108-109.
[9] BIANCHI E,BONAZZA A,MARSANO E.Free radical grafting onto cellulose in homogeneous conditions.1.Modified cellulose-acrylonitrile system[J].Cabrohydrate Polymers,1998,36(4):313-318.
[10] 许冬生.纤维素衍生物[M].北京:化学工业出版社,2001:20-21. XU Dong-sheng.Cellulose Derivativies[M].Beijing:Chemical Industry Press,2001:20-21.
[11] 熊磊,于伟东.酸处理后纤维素分子结构的显微红外光谱分析[J].纤维素科学与技术,2013,21(2):60-61. XIONG Lei,YU Wei-dong.Analysis of the cellulose macromolecule structure after acid treatment by FTIR microspectroscopy[J].Journal of Cellulose Science and Technology,2013,21(2):60-61.
[12] CHEN Jing,ZHANG Jin-ming,FENG Ye,et al.Synthesis,characterization,and gas permeabilities of cellulose derivatives containing adamantane groups[J].Journal of Membrane Science,2014,469(1):507-514.
[13] 骆微,李丽琴,孙高穹,等.微波辅助离子液体法对纤维素的均相改性研究[J].纤维素科学与技术,2014,22 (4):12-17. LUO Wei,LI Li-qin,SUN Gao-qiong,et al.Homogeneous modification of cellulose by microwave assisted ionic liquid[J].Journal of Cellulose Science and Technology,2014,22(4):12-17.
[14] 宁永成.有机波谱学谱图解析[M].北京:科学出版社,2010:111-114. NING Yong-cheng.Spectrum Analysis of Organic Compounds[M].Beijing:Science Press,2010:111-114.
[15] 莫亚莉,刘娟,陈日清,等.丙烯酸纤维素的制备与表征[J].林产化学与工业,2013,33 (3):74-77. MO Ya-li,LIU Juan,CHEN Ri-qing,et al.Synthesis and characterization of acrylate cellulose[J].Chemistry and Industry of Forest Products,2013,33(3):74-77.
[16] 陈立仁.液相色谱手性分离[M].北京:科学出版社,2006:280-281. CHEN Li-ren.Chiral Separations by High Performance Liquid Chromatography[M].Beijing:Science Press,2006:280-281.