摘要
羟基脯氨酸正十二烷基酯同时含有亲水和疏水两种基团,具有良好的表面活性,改变了羟基脯氨酸与脂不相溶无法通过细胞膜的缺点,可以将羟基脯氨酸顺利带进细胞。同时,由于其含有一个羟基,为进一步反应提供了条件,可以作为其他药物以及化工产品的中间体。
本文以L-羟基脯氨酸和正十二醇为原料,对甲基苯磺酸为催化剂,甲苯为带水剂合成羟基脯氨酸正十二烷基酯。催化剂用量、合成反应时间、反应温度和底物配比均影响羟基脯氨酸转化率。L16(45)正交实验结果指出,合成羟基脯氨酸正十二烷基酯的最佳工艺条件为:反应时间10小时,温度130℃,羟基脯氨酸与正十二醇的摩尔比为1:5,羟基脯氨酸与对甲基苯磺酸的摩尔比为1:1.08。在该最优条件下合成羟基脯氨酸正十二烷基酯,羟基脯氨酸的转化率为85.57%。各因素所选的各水平内,对羟基脯氨酸转化率影响因素的大小顺序为底物配比>反应温度>反应时间>催化剂用量。
分别采用结晶法和硅胶柱层析法对羟基脯氨酸正十二烷基酯进行分离纯化,结晶法的优点就是操作简单,速度较快,在纯度要求不高时可采用该法。硅胶柱层析法所得的产物,不论是纯度还是收得率都比结晶法高,所以柱层析法分离纯化产物是一个较优的方法。结晶获得羟基脯氨酸正十二烷基酯纯度为89.46%,收率为78.62%;硅胶柱层析所得纯度为90.17%,收率为87.34%。将二次层析后的羟基脯氨酸正十二烷基酯制备成冻干粉,进行红外和核磁共振分析和结构表征,确定获得的高纯度产物为羟基脯氨酸正十二烷基酯。
羟基脯氨酸上有羧基,仲羟基和亚氨基三个亲水性基团,极易与水分子形成氢键而溶解在水中。当其成酯后其最强的亲水性基团羧基被长链烷基取代,亲水性降低;同时长链烷基的位阻作用阻碍仲羟基和亚氨基与水分子作用,最终造成在水中的溶解度大幅下降,脂溶性增强。其油/水分配系数从-1增加到0.5。
羟基脯氨酸正十二烷基酯的仲羟基和亚氨基可以作为氢供体,具有提供质子的能力,使强氧化性的超氧阴离子(O2-·)还原,从而终止自由基链式反应,起到清除超氧阴离子(O2-·)的作用;同时,能与金属离子形成螯合物,从而阻止Fenton反应的发生,进而阻止羟基自由基(·OH)产生。当加入酯的浓度为10 mg/mL时,对O2-·的清除率达到了69.3%,对·OH的清除率为67.62%。
Hydroxyproline lauryl alcohol ester comprises hydrophilic and hydrophobic groups both, the surfactivity of which is excellence. The disadvantage that hydroxyproline is indiscerptible in fat and can not get through the cell membrane is changed by esterification to enter the cells favoringly. At the same time, Hydroxyproline lauryl alcohol ester could be used for intermediate of medicament, because of the dissociative hydroxyl.
In this article, L-hydroxyproline and lauryl alcohol were used as materials of esterification, paratoluene sulfonic acid was used as catalyzer, and toluene was used as the water schlepped. Dosage of catalyzer, time and temperature of reaction, scale of materials affecte the reactive rate of hydroxyproline. It is showed by the results of L9 (43) orthogonal tests that the optimum conditions are that the time of reaction was 10 hours; the temperature of esterification is 130℃;the molar scale of hydroxyproline and lauryl alcohol is 1:5; the molar scale of hydroxyproline and paratoluene sulfonic acid is 1:1.08. With the optimum conditions, the reactive rate of hydroxyproline is 85.57%. The effect of these factors is different that the molar scale of hydroxyproline and lauryl alcohol > temperature > time > molar scale of hydroxyproline and paratoluene sulfonic acid.
Crystal and silica gel column chromatography were used for purification of hydroxyproline lauryl alcohol ester. The advangtage of crystal are simply operation and fast separation, because of which crystal is used for getting low purity. Because the purity and the yield by silica gel column chromatography were both higher than crystal, silica gel column chromatography was chose to purify hydroxyproline lauryl alcohol ester in this article. The purity and yield of hydroxyproline lauryl alcohol ester were 89.46% and 78.62% by crystal, and 90.17% and 87.34% by silica gel column chromatography respectively. After silica gel column chromatography twice, hydroxyproline lauryl alcohol ester was prepared to freeze-dried powder, and was characterized the structure by IR and NMR.
There are three hydrophilic group: carboxyl, -OH and–NH2 on the hydroxyproline, which form hydrogen bond with water and dissolve in water. Because of that the carboxyl is replaced with long-chain alkyl, hydrophilic capability of hydroxyproline is reduced; and the steric effect of long-chain alkyl blocks the effect of -OH and–NH- with water, that the solubility reduces significantly in water, but increases in fat. The oil-water partition coefficient increase from -1 to 0.5.
The -OH and -NH- of hydroxyproline lauryl alcohol ester can be used for hydrogen donor, and deoxidize O2-·; hydroxyproline lauryl alcohol ester can form chelate compound with metallic ion, and stops the Fenton reaction, then stops the appearance of·OH. When the concentration of hydroxyproline lauryl alcohol ester is 10 mg/mL, the rate of O2-·elimination is 69.3%, and·OH elimination was 67.62%.
引文
[1]沈同.生物化学(上册) [M].北京:高等教育出版社,1985:1113-114.
[2]吴显荣.氨基酸营养[J].氨基酸杂志. 1988, (2) :21-28.
[3]肖小年,马永花,易醒,刘翠,胡少华.六氢β-酸的抗氧化功能研究[J].江西农业大学学报.2008,30(3):517-520.
[4]汤新之,崔乃杰.临床生物化学[M].天津:天津科学技术出版社,1999:719-723.
[5]孙亚捷,李新芳.南沙参多糖对小鼠的抗衰老及清除氧自由基作用研究[J].中国药师.2005年8(9).
[6]周燕,欧阳安力,华平.抗氧化剂对皮肤角质细胞体外寿命的影响[J].生物工程学报. 2002, 18(5): 630-633.
[7] Michael Lam, Maria Sulindro. Aging skin [R]. Anti -Aging Research Brief (A3R) , 2001.
[8]左瑞雅,周小华杜首英. L-羟脯氨酸-Zn(Ⅱ)的配合机制及其抗氧化性研究[J].生物工程学报.2007,23(4):704-709.
[9]上海医药工业研究院主编.生化药物·药品集(第七分册) [M ].上海:上海科学技术出版社, 1984: 56.
[10]范镇基.非蛋白氨基酸的功能与应用[ J ].氨基酸杂志.1990,1.
[11] 14氨基酸注射液.;上海长征制药厂[ R ];医药工业.1984, 1.
[12]上海长征制药厂; 14氨基酸注射液;医药工业[ J ]; 1983; 6 .
[13]吴显荣.氨基酸营养[ J ].氨基酸杂志.1988: 2-3.
[14]刘仁涌,李慧琴,刘春庆,付君.卡托普利合成方法的改进[J].沈阳药科大学学报. 1997,14(4).
[15]缪正兴,张仲明.L-脯氨酸的生产及其应用[J] .发酵科技通讯,2004 ,33(2) :14 -16.
[16]王炳琴,黎植昌.氨基酸酯的制备方法及应用[J].氨基酸和生物资源. 1995. 17 (4) : 40-45.
[17]周晓辉,叶小薇.甲基葡萄糖苷烷氧基化合物的合成与应用[J].化学与粘合. 1998, 17 (3) : 165-166.
[18]左川幸一郎.氨基酸系表面活性剂[J].日用化学工业译丛. 1989, (1) : 1-5.
[19]上村洋一.氨基酸及其衍生物对头发的作用和效果[J].日用化学工业译丛. 1988, (4): 44-46.
[20]田伟.高级烷酯杀菌剂[J].农药. 1988, 2: 23-25.
[21]尚岩,王春颖.氨基酸酯合成方法最新研究进展[J].哈尔滨理工大学学报. 2008,13(2).
[22] James R, et.al.. The preparation of optically active pepiteds using mixed carbonic-carboxylic acid anhydrides [J]. Am. Chem. SOC., 1953, 75 : 5556.
[23] Tadashiro, et.al.. Synthesis 3,6-bis (α-hydroxyethyl)-2,5(dioxopiperazine). Ball. Chem. SOC. Japan. 1966 : 39 (7): 1598-1599.
[24] Howard Sachs, Erwinb. Benzyl Esters of Glutamic Acid [ J ].J. Amer. Chem. Soc. , 1953, 75 (18) : 4610– 4611.
[25] Akopla,M, Higuchi, C. Preparation and Isolation ofMineralAcid Salt of an Amino Acid Methyl Ester [ P ]. EP:0376230A2,1989 - 12– 24.
[26]冯光炷,朱春山,王晓君,谢文磊.复合氨基酸月桂醇酯的合成及性能[J].郑州工程学院学报. 2003, 24(4) : 42-45.
[27]张菊娴,张天太.甲酸丙酯和甲酸异丙酯的催化合成研究[J].化学工程师.1991,2:18.
[28] Michiko Watanabe, Atsuko Shimada, Etsuko Yazawa, Tamami Kato, Soichi Arai. Proteinaceous Surfactants Produced from Gelatin by Enzymatic Modification : Application to Preparation of Food Items [J]. Journal of food science, 1981, 46 : 1738.
[29] Roger R. Preparation of t - Butyl Esters of Free Amino Acids[ J ]. J. Org. Chem. , 1963, 28 (5) : 1251 -1253.
[30] Akintola. A, Abodering. R, Joseph. S. Fruton Benzhydryl Esters of Amino Acids in Pep tide Synthesis[ J ]. J. Amer. Chem.Soc. , 1965, 87 (23) : 5469-5472.
[31]安广杰,王璋,许时婴.蛋氨酸烷基酯的合成[J].食品工业科技算-食品添加剂. 2004, 12 : 107 -117.
[32] Tadashiro. et. a1.. Synthesis 3,6-his( hydroxyethy1)-2,5(dioxopiperazine). Ball. Chem. SOC. Japan. 1966, 39 (7) : 1598-1599.
[33] Ravindra. P. Patel. S P. Synthesis of Benzyl Esters of a - A2mino Acids[ J ]. J. Org. Chem. , 1965, 30 (10) : 3575-3576.
[34]思洋,刘乐乐,张廓,姜萍萍.氨基酸甲乙酯的合成及纯化[J].内蒙古医学院学报. 2005, 27 (1) : 33-34.
[35] RAMESH C, ANAND V. A Mild and Convenient Procedure for the Esterification of Amine Acid [ J ]. Synthetic Commun. 1998,28 (11) : 1963 -1965.
[36] Zander. N, Frank. R. Polystyrylsulfonyl Chloride Resin: an Efficient Solid-supported Condensation Reagent for the Solution Phase Synthesis of Esters [ J ]. Tetrahedron Lett, 2001, 42 (44) :7783 -7785.
[37] Sieber.P. An Improved Method for Anchoring of 9-fluorenylme2-thoxycarbonyl-Amino Acids to 4-Alkoxybenzyl Alcohol Resins.[ J ]. Tetrahedron Lett. , 1987, 28 (49) : 6147-6150.
[38] Blankemeyer-Menge B, Nimtzm, Frank. R. An Efficient Method for Anchoring Fmoc- anino -Acids to Hydroxyl2Function2alised Solid Supports [ J ]. Tetrahedron Lett. 1990, 31 ( 12 ) :1701 -1704.
[39] Norbert. Z, Jurgen. G, Ronald. F. Polystyrylsulfonyl-3– nitro-1H-1, 2, 4-triazolide- resin: a New Solid-supported Reagent fort he Esterification of Amino Acids[ J ]. Tetrahedron Lett. , 2003,44: 6557 -6560.
[40]邓斌,黄海英,李强国,等. SnCl4. 5H2O /C催化合成谷氨酸月桂酯[ J ].合成化学, 2005, 13 (2) : 208 -210.
[41] Matthial. PM,Dietr Ichm. A., Dieter. B A. . Processs for Preparing Amino Acid Esters and Their Acid Addition Salts [ P ]. US: 2005 /0192460 A1, 2005 - 09– 01.
[42]卓仁禧,刘高伟,彭普平.5 -氟尿喃咤‘一甲酸基氨基酸、短肤的合成及抗肿瘤活性[J].高等学校化学学报.1991,12(4):555-559.
[43] Gedye R, Smith F, Westaway K, etal. The use of microwave ovens for rapid organic synthesis [J]. Tetrahedron Lett, 1986, 27(3):278-280.
[44] Vasanthakumar GR, Patil BS, Babu VVS. Microwave Assisted Facile Synthesis of Amino Acid Benzyl Ester p– toluene sulfonate and Hydrochloride Salts [ J ].Letters in Pep tide Science, 2002, 9: 207 -209.
[45] Zhao. H, Sanjay. V M. Esterification of Amino Acids by Using Ionic Liquid as a Green Catalyst[ J ]. Chemical Industries (Dek2ker) , 2003, 89: 667- 672.
[46] Kise. H, Shirato. H. Synthesis of Aromatic Amino Acid Ethyl Esters byα-chymotryp sin in Solutions of High Ethanol Concentra2tions [ J ]. Tetrahedron Lett. , 1985, 26 (49) : 6081- 6084.
[47] Shu. I-Tein C, Kung TsungWang. Papain Catalysed Esterification of N - Protected Amino Acids [ J ]. Chem. Commun. , 1988 (4) 327-329.
[48] Yesiloglu Y, Kilic I. Polyvinyl Alcohol-Trypsin as a Catalyst for Amino Acid Ester Synthesis in Organic Media [ J ]. Preparative biochemistry & biotechnology, 2004, 34 (4) : 365-375.
[49]汪茂田,谢培山,王忠东,王海棠、吴云骥、周海梅等编著.天然有机化合物提取分离与结构鉴定[M].北京.化学工业出版社.2004:5-200.
[50]张克胜,穆林静,龙韫先.α-吡唑基-N -苯基-α-氨基膦酸酯的合成及生物活性的初步研究[J].高等学校化学学报.1999,20(5):741-743.
[51]蒋庆锋,周有骏,姚斌,郑灿辉,吕加国,朱驹. 9-芴甲氧羰基氨基酸叔丁基酯的合成[J].第二军医大学学报.2007,28(1):101-102.
[52]黄锁义,王麟生.N-Boc-3-碘锌-L-丙氨酸甲酯的合成研究[J].化工技术与开发. 2004,33(2):13-16.
[53]王海英,杨建洲.硬脂酰取代二羟乙基甘氨酸酯的合成及在造纸施胶中的应用研究[J].造纸化学品.2005,2:25-28.
[54]鄢立刚,王捷,谢怀江,徐柄祥,姚斌.N-碘炔丙基氧羰基氨基酸及其酯类化合物的合成和抗菌活性的研究[J].中国医药工业杂志.1999,30(7):305-307.
[55] J W Jorgenson,K D Lukacs.Zone electrophoresis in open tubular class cayillaries[J].Anal Chem,1981,53:1298.
[56] S Terabe,K Otsuka,K Lchikaws,et al.Electro kinetic separations with micellarsolutions and open tubular capillares[J].Anal Chem,1984,56:111.
[57] S Hjierter,J Liao,K Yao.The oretical and experimental study of high performance electro- phoretic mobilization of iscele critically focused peotein zones[J].J Chromatogr, 1987, 387:127.
[58] A Chohen,B Karger.High performamce sodium dodecyz Prolyacrylamide gel vapilay electro- phorzsis of peptides and prote ins[J].J romatogr 1987,397:409.
[59]卢楠,姜艳军.毛细管电泳及其在分离分析中的应用进展[J].河北化工. 2005,5:24-26.
[60] Fabienne F , Olivier P. Synthesis and applications of the first polyfluorous praline derivative [J]. Tetrahedron : Asymmetry, 2003, 14:139-143.
[61] Kwon HY,Langer R. Pseudopoly (amino acids): a study of the synthesis and characterization of poly (trans-4-hydroxy-N-acyl-L-proline esters ) [J] . Macromolecules, 1989, 22:3250-3255.
[62] Lee RS,Yang JM. Synthesis and Characterization of Degradable Copoly (trans-4- hydroxy-L-proline/ε-caprolactone) [J]. Journal of Applied Polymer Science, 2003, 88: 3176-3182.
[63] Lee RS,Lin TF, Yang JM. Synthesis and Characterization of Novel Poly(estercarbonate)s by Copolymerization of Trans-4-hydroxy-L-proline with Cyclic Carbonate [J]. Journal of Polymer Science:Part A:Polymer Chemistry, 2003, 41:1435-1443.
[64] Lee RS,Lin TF ,Yang JM. Novel, Biodegradable, Functional Poly(ester-carbonate)s by Copolymerization of Trans-4-hydroxy-L-proline with Cyclic Carbonate Bearing a Pendent Carboxylic Group [J]. Journal of Polymer Science:Part A : Polymer Chemistry, 2004, 42 : 2303-2312.
[65]王文刚,挥榴红,王睿,周筱青.经皮吸收促进剂的研究与应用进展[J].中国新药杂志.2006,15(4).
[66]何建平.药物经皮吸收的物理促渗方法[M].江西医药.2004,39(6).
[67]韩璐,胡晋红,朱全刚.经皮给药系统促渗方法研究的新进展[M].中国新药杂志.2007,16(4).
[68]昝佳,蒋国强,陈军,马建国,丁富新.经皮给药系统的研究进展和发展趋势[J].精细化工.2002, 19(6).
[69] Beall H, Sloan KB. Int J Pharm.1996,129 :203.
[70]谢晶曦,常俊标,王绪明.红外光谱在有机化学和药物化学中的应用(修订版)[M].北京:科学出版社,2001:37-67.
[71]宁永成.有机化合物结构鉴定与有机波谱学(第二版)[M].北京:科学出版社,2000:1-26.
[72]王连生,韩朔睽,孔令仁.分子结构、性质与活性[M] .北京:化学工业出版社,1997. 26-41.
[73] Michniak BB , Player MR , Godwin DA et al . A study of enhancer structure activity relation- ships in a series of L - oxopiperidine - 1 - acetic acid esters. Pharm Res ,1995 ,13(9) : 260-268.
[74] Siddhuraju P,Becker K. The antioxidant and free radical scavenging activities of p rocessed cowpea seed (V igna unguiculata (L. ) Walp. ) extracts[ J ]. Food Chem , 2007, 101 (1) : 10-19.
[75]张德莉,罗光富,陈燕,黄应平.溴甲酚绿测定Fenton产生的羟基自由基[M].2005,27(5): 449-452.
[76]宁正祥.食品成分分析手册[M].北京:中国轻工业出版社, 1998:174-196.
[77]覃兆海,金淑惠,李楠.基础有机化学[M].北京:科学技术文献出版社,2004:342-363.
[78]磨建新.温度对化学反应速度、平衡与转化率的影响[J].高教论坛,2004,10(5):162-164.
[79]王璋.食品化学[M].北京:轻工业出版社, 1999:140-149.
[80]王英姿,任天池,孙秀梅.高乌甲素的油水分配系数及透皮吸收速率常数的相关性研究[J].山东中医药大学学报. 2006,30(2):155-156.
[81] H. van de, H. van de Waterbeemd. (何仲贵,钟大放译).药物生物利用度[M].北京:化学工业出版社,2007:3-20.
[82]王培林.医学细胞生物学[M].北京:人民卫生出版社, 2005:65-82.
[83]陈瑗,周玫.药用化学[M].北京:人民卫生出版社, 2002 :8 -201.
[84]谢梅林,顾振伦,钱曾年.槲皮素的抗超氧阴离子作用[J].现代应用药学,1992,9(1):1-3.
[85]张强,阚国仕,陈红漫.玉米抗氧化肽的分离制备及其体外抗氧化活性的研究[J].中国粮油学报,2005,20(5):36-39.