三肽纤维素酯的制备及应用研究

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。

Preparation and Application of Tripeptide-cellulose Ester

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 ( DS ) 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 DS 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.