环糊精的药学应用

环糊精是一类由D-吡喃型葡萄糖单体以α-1,4-糖苷键相互连接而成的环状低聚糖。这种环状的低聚糖形成了一种内穴疏水、外缘亲水的特殊结构。因此,环糊精能够和脂溶性药物分子形成包合物,从而可以增加脂溶性药物分子的水溶性和稳定性。同时,这些低聚糖自身无毒性,亦没有明显的药理活性。这些使得环糊精被广泛应用于食品、化妆品和药品当中。作为一种新型的药物赋形剂,环糊精广泛应用于口服、静脉注射和鼻下给药等多种药物剂型,目前在市场上至少有35种环糊精相关的药品,相信在不久的将来,环糊精在药学领域会得到越来越广泛的应用。本文综述了环糊精类的结构特点、包合性能、药物代谢、毒理学性质和目前在药学领域的应用。     

作为非病毒基因载体的环糊精及其衍生物

环糊精由于自身的生物相容性和结构易裁剪性,通过结构修饰、聚合或超分子组合等设计被逐渐应用于非病毒基因载体系统。本文将分别从环糊精、其小分子衍生物、含环糊精聚合物以及超分子结构综述国内外近几年的设计思路和研究进展,并探讨含环糊精及其衍生物的非病毒基因载体的"结构-安全性-基因转染效率"关系。     

环糊精葡萄糖基转移酶的结构特征与催化机理

随着环糊精在食品、医药等领域的应用越来越广,生产环糊精所必需的环糊精葡萄糖基转移酶(CGT酶)已经成为当今研究的热点。特别是近二十年来,国外对该酶进行了比较深入的研究。首先介绍了CGT酶的功能特性与结构特征。CGT酶是一种多功能型酶,能催化三种转糖基反应(歧化、环化和耦合反应)和水解反应,其中,能将淀粉转化为环糊精的环化反应是特征反应;作为α-淀粉酶家族的成员,CGT酶除了具有与α-淀粉酶相同的A、B、C结构域外,还存在D和E结构域。另外,对CGT酶的催化机理包括底物结合方式、转糖苷反应机理以及环化机理等进行了详细的讨论。     

环糊精葡萄糖转移酶的研究进展

环糊精葡萄糖转移酶在工业上用于生产环糊精.它可通过环化作用将淀粉转化成环糊精.由于环糊精具有一个亲水的外表面和一个疏水的内核,疏水的内核能包上非极性物质而溶于水中,所以在食品、化妆品、药品领域被广泛使用.综述环糊精葡萄糖转移酶近年在基因工程及蛋白质工程等方面的研究发展状况.     

亚位点?7处突变对碱性芽胞杆菌CGT酶产物特异性的影响

为了研究来源于碱性芽胞杆菌的γ-环糊精葡萄糖基转移酶(CGT酶)具有较高产物特异性的作用机理,对其氨基酸序列和模拟结构进行了分析,确定其亚位点7处氨基酸的缺失可能影响其产物特异性。运用重叠PCR的方法,在其亚位点7处添加缺失的6个氨基酸,造成插入突变。将突变基因与pET-20b(+)连接并在大肠杆菌BL21(DE3)中表达。以可溶性淀粉为底物进行酶转化,HPLC分析转化产物中的环糊精含量。结果表明,相对于野生型γ-CGT酶,突变酶转化生成的3种环糊精中,γ-环糊精所占的比例从76.0%降至12.5%,α-、β-环糊精分别从8.7%和15.2%提高至37.5%和50%。分析其可能机理为:与α-、β-CGT酶相比,野生型γ-CGT酶的亚位点7处缺失6个氨基酸,该构象为葡萄糖的结合提供了更大的空间,从而更适合γ-环糊精的生成;而在其亚位点7处插入6个氨基酸,造成插入突变后,葡萄糖链结合的空间变小,这种构象不利于γ-环糊精的生成。     

利用高效阴离子色谱快速检测筛选环糊精糖基转移酶产生菌>

利用高效阴离子色谱快速直接地检测微生物发酵液中的环糊精成分,尤其是大环环糊精的组成,进而创造了一种能快速准确地从土壤中筛选产环糊精糖基转移酶菌种的方法。共分离了149个产胞外淀粉水解酶的微生物菌株,利用高效阴离子交换色谱共检测了其中11株菌,其中6株主要产CD6 ,5株主要产CD7,主要产CD8的没有。在直接鉴定产生环糊精糖基转移酶菌株的过程中,也可以定量检测各种环糊精包括大环糊精（CD大于8）的含量。     

β-环糊精主客体识别抗菌剂及抗菌性能研究进展

细菌污染与人类的生存息息相关,随着多重耐药细菌的增加,提高抗菌剂抗菌性能、抑制细菌传播和感染成为抗菌剂研究领域的热点问题。利用β-环糊精的包结作用与抗菌剂主客体识别改善抗菌剂的理化性质,增强抗菌剂的抗菌性能引起了广泛关注。环糊精具有内疏水外亲水的特殊结构,疏水内腔可选择性地与空腔大小相匹配的疏水客体分子进行主客体识别,将抗菌剂包合,从而改善抗菌剂理化性质,提高抗菌剂抗菌性能。本文综述了β-环糊精及其衍生物与有机抗菌剂、天然抗菌剂主客体识别对抗菌剂理化性质及抗菌性能的影响,分析了主客体识别的包合机理及主客体之间的作用,系统地归纳总结了环糊精主客体识别抗菌剂对不同细菌的抗菌作用及在医药、纺织、食品等领域的应用研究现状,以期为环糊精主客体识别抗菌剂的深入研究及应用提供参考。     

环糊精葡萄糖基转移酶的性质、应用与固定化研究进展

本文总结了环糊精葡萄糖基转移酶的性质、应用与固定化研究的进展情况,引用文献４７篇。     

基于分子动力学模拟的高γ-环糊精专一性环糊精糖基转移酶理性改造

【背景】环糊精糖基转移酶的分子动力学模拟较传统基因改造而言能有效提高改造效率,减少盲目性。【目的】探究环糊精糖基转移酶的催化专一性机理,为获得产γ-环糊精专一性更高的环糊精糖基转移酶提供高效突变菌株方法。【方法】通过分子对接和分子动力学模拟,获得3种产物类型CGTase与底物的对接模拟结构,并通过定点突变实验进行验证。【结果】分子动力学模拟结果显示α-和β-CGTase与十糖链在酶蛋白S1区域呈现闭合的形态,而γ-CGTase和十糖链在S1区域呈现更易于生成γ-环糊精的张开形态;3种CGTase与十糖链在相同位置存在氢键的氨基酸共有17个相对应位点,其中14个位点的氨基酸种类一致,不一致的3个氨基酸对应α-CGTase位点分别为Y89、D234和Y262。本研究对Y262位点进行定点突变和产物专一性实验,结果显示经过分子动力学预测的Y262L有助于提高产γ-CD专一性,从野生酶的13.7%提高到39.9%,γ-环糊精产物比例提高了3倍。【结论】分子动力学模拟结果对于指导环糊精糖基转移酶的专一性内在机理具有一定的正向指导意义。     

β-环糊精及其衍生物在生物制药领域中的应用

β-环糊精及其衍生物近年来一直是人们研究的热点,研究人员尝试将其应用到制药、食品、化工和农业等领域。本介绍了β-环糊精及其衍生物的结构和理化性质,并综述了它们在生物制药领域中的应用,重点介绍了在生物药物制剂方面的进展。     

Role of capillary electrophoresis methods in the drug development process

In the past several years, capillary electrophoresis (CE) has generated considerable interest from pharmaceutical companies for control of both the chiral and achiral purity of bulk drugs and drug products. This paper evaluates the use of CE as: (1) a technique complementary to HPLC for the determination of peak homogeneity of a drug, (2) for determination of chiral purity, and (3) for determination of achiral purity. It would be greatly advantageous if CE could be used to determine both the chiral and achiral purity in a single assay. This investigation compares the results obtained for the separation of the enantiomers of duloxetine using several neutral cyclodextrins to those obtained using anionic cyclodextrins (sulfobutyl ether derivatives) as chiral selectors added to the separation buffer. In addition, it reports chiral separations obtained by using neutral cyclodextrins in a sulfonic acid-coated capillary column, which give a negatively charged capillary surface and electro-osmotic flow even in low pH buffers. The possible mechanism of separation is discussed. © 1996 Wiley-Liss, Inc.     

Epididymal sperm profiles in young adult,middle-aged,and testosterone-supplemented old rats

Both ejaculated semen and epididymal contents from an individual male contain sperm that differ in various physicochemical characteristics. An experiment is reported in which epididymides from rats 5–24 months old were subjected to density gradient centrifugation to separate gametes of different stages of maturity. The research was designed to examine typical changes in “profiles” of sperm maturity during the reproductive lifetime of rats. Also, testosterone complexed with cyclodextrin that mimics the episodic release of the endogenous hormone was used to supplement the decreased circulating titers of some of the old males. Results revealed clear ontogenetic patterns of gradually decreasing reproductive competence as measured by absolute numbers of sperm, circulating levels of testosterone, and various other physiological markers of fertility. Sperm profiles also revealed age-specific changes with a shift toward progressively more mature, perhaps senile, gametes that begins at middle age. Testosterone supplementation (400 μg/kg b.w./day for 30 days) failed to restore sperm numbers or other measures of physiology in the old males, but the steroid modified sperm profiles to approximate more closely the profiles characteristic of young adult males than either untreated middle-aged or old males. The data were interpreted as suggesting that epididymal sperm profiles clearly identify males of different ages, and that the aging epididymis retains its capacity to respond to manipulations that modify the endocrine milieu.     

Complexation of Tocotrienol with γ-Cyclodextrin Enhances Intestinal Absorption of Tocotrienol in Rats

To determine the bioavailability of tocotrienol complex with γ-cyclodextrin, the effects of tocotrienol/γ-cyclodextrin complex on tocotrienol concentration in rat plasma and tissues were studied. Rats were administered by oral gavage an emulsion containing tocotrienol, tocotrienol with γ-cyclodextrin, or tocotrienol/γ-cyclodextrin complex. At 3 h after administration, the plasma γ-tocotrienol concentration of the rats administered tocotrienol/γ-cyclodextrin complex was higher than that of the rats administered tocotrienol and γ-cyclodextrin. In order to determine the effect of complexation on tocotrienol absorption, rats were injected with Triton WR1339, which prevents the catabolism of triacylglycerol-rich lipoprotein by lipoprotein lipase, and then administered by oral gavage an emulsion containing tocotrienol, tocotrienol with γ-cyclodextrin, or tocotrienol/γ-cyclodextrin complex. The plasma γ-tocotrienol concentration of the Triton-treated rats administered tocotrienol/γ-cyclodextrin complex was higher than that of the other Triton-treated rats. These results suggest that complexation of tocotrienol with γ-cyclodextrin elevates plasma and tissue tocotrienol concentrations by enhancing intestinal absorption.     

Production of cyclodextrin from starch by cyclodextrin glycosyltransferase from Bacillus firmus and characterization of purified enzyme

During screening for cyclodextrin-forming microorganisms, an alkalophilic Bacillus sp, which produced high activity of cyclodextrin glycosyltransferase, was isolated and identified as Bacillus firmus. The crude enzyme transformed starch to mainly β-and γ-cyclodextrin. The purified enzyme had an optimum pH of 7.5–8.5 and its optimum temperature was 65°C, which is the highest optimum temperature as compared to other cyclodextrin glycosyltransferases except that produced by Bacillus amyloliquefaciens. Received 06 January 1997/ Accepted in revised form 20 March 1997     

Evaluation of enantiomeric purity of selected amino acids in honey

Highly sensitive and accurate HPLC methods were used for the determination of total amounts of proline, leucine and phenylalanine and their enantiomeric ratios in a variety of different honey samples. Significant amounts of D -leucine and D -phenylalanine and relatively low concentrations of D -proline have been found in honeys of different botanical and geographical origins. It is suggested that the enantiomeric ratios of amino acids could be used to test for storage effects, age, and the quality of the processing of the honey. © 1994 Wiley-Liss, Inc.     

Effects of k-carrageenan supplementation or in combination with cholesterol-loaded cyclodextrin following freezing-thawing process of rooster spermatozoa

This experimental research purposely seeks to explore the effect of supplementing k-carrageenan (k-CRG) or CLC (cholesterol-loaded cyclodextrins) or the combined effect of k-CRG and CLC as supplements of antioxidants to an extender for rooster semen freezing. A total of 75 neat pooled ejaculates were collected twice a week from twenty-five (25) commercial line arbor acres broiler roosters (30 wks) during the experimental period. In each replicate, semen samples (n= 15, three ejaculates per rooster) were pooled and divided into nine equal aliquots, and each aliquot was diluted with one of the following extender supplemented with k-CRG, CLC, and k-CRG + CLC after which it was subjected to cryopreservation process using the “pellet” method. In study I, the supplementation of extenders with k-CRG was in five equal aliquots as follows; (0.2, 0.4, 0.6, 0.8) mg/mL and control group (k-CRG 0) mg/mL while in Study II, there was a combination of both k-CRG + CLC (0.4 mg/mL + 1.5 mg/mL, respectively), 0.4 mg/mL k-CRG, 1.5 mg/mL CLC and control group. Sperm quality parameters, endogenous antioxidant enzymes, lipid peroxidation (MDA) and ROS were all assessed after the freeze-thaw process. Our findings in study I indicated that at post-thaw, an optimum 0.4 mg/mL k-CRG supplementation in the extender improved semen quality parameters, endogenous enzymes, MDA and ROS in comparison to the control group. Interestingly prior to the freeze-thaw process, it was depicted in study II that combined k-CRG + CLC (0.4 mg/mL+1.5 mg/mL) inclusion in the extender provided maximum protection to sperm quality parameters, endogenous enzymes, MDA and ROS in comparison to 1.5 mg/mL CLC and control group at post-thaw. Besides, there was also a significant difference observed in the extenders supplemented with combined k-CRG + CLC (0.4 mg/mL +1.5 mg/mL) when compared to 0.4 mg/mL k-CRG for semen quality parameters and endogenous antioxidant enzymes (SOD, CAT, and GPx) but no significant difference was observed for MDA and ROS. Also, there was a significant difference observed in the extender supplemented with 1.5 mg/mL CLC when compared to the control group for semen quality parameters, SOD, CAT, and MDA but no significant difference for GPx and ROS at post-thaw. In conclusion, k-CRG at an optimal dosage of 0.4 mg/mL proved effective for improving post-thaw sperm quality but its combined addition k-CRG + CLC at an optimal concentration of (0.4 + 1.5) mg/mL in the extender provided greater protection to the rooster spermatozoa at post-thaw.     

Reverse Enrichment in a Multi-stage Bubble Separator

An unexpected phenomenon named “reverse enrichment” was observed, in which, contrary to the normal behaviour, isoelectric hemoglobin coagula concentrated in the bottom stage of a multistage bubble separator. The phenomenon was observed when the hemoglobin particles were highly concentrated, the bubble diameter was larger than one third of the hole diameter of the partition plate, and the superficial air velocity at the hole was high. Transient and final behaviours of the multi-stage bubble separator with the reverse enrichment were analyzed by assuming the rejection of particles from upward thin liquid streams.