生物活性肽SSDI固相合成工艺的研究

目的:研究生物活性肤SSDI的固相合成工艺,并为大规模合成目标肽提供理论依据.方法:采用固相合成法,原料氨基酸以Fmoe形式保护,用Wang树脂为载体,经1-氧3-双二甲胺羰基苯骈三氮唑四氟化硼盐(TBTU)\1-羟基-苯并-三氮唑(HOBt)\二-异丙基乙胺(DIEA)混合试剂缩合,20%哌啶的DMF溶液脱保护,用三氟乙酸\茴香硫醚\巯基乙醇\苯酚\水混合作为切割试剂将多肽从Wang树脂上切割下来.结果:多肽粗品的得率高达70%,经RP-HPLC纯化,可获得纯度在98%以上的目标肽,经MALDI-MS质谱鉴定其分子量与理论值一致.结论:此合成方法操作简单,产品得率高,适合大规模合成目标肽.     

两种树脂固相合成机体保护肽的比较

目的:比较Wang树脂和二氯三苯甲基树脂(以下简称二氯树脂)在固相合成机体保护肽中的表现,为工业化生产提供理论依据。方法:通用的Fmoc固相合成法制备机体保护肽,通过比色法对两种树脂与氨基酸的偶联率进行比较,用质谱(MALDI-TOFMS)对粗肽进行鉴定分析,用反相高效液相色谱(RP-HPLC)进行粗肽的分析和纯化。结果:显示以二氯树脂作载体,第一个氨基酸的连接率,以及机体保护肽的纯度和产率都要明显高于Wang树脂。结论:二氯树脂作载体合成机体保护肽更适合大规模工业化生产的要求。     

人胰岛素原C肽固相合成的改进

利用谷氨酸的γ羧基与 4 甲基二苯甲基胺 (MBHA)树脂的氨基偶联固相 ,合成经HF切割去保护基后形成C端是谷氨酰胺的人胰岛素原C肽。MBHA树脂相对价格便宜 ,此方法是制备人胰岛素原C肽的另一种尝试。同时 ,也用PAM树脂合成了人胰岛素原C肽类似物。     

Fmoc固相合成JFT的工艺研究

目的:研究多肽JFT的合成工艺。方法:本实验采用固相合成法(spps),以Fmoc—氨基酸为原料,TBTU\HoBt\DIEA混合试剂缩合,用三氟乙酸\苯甲硫醚\巯基乙醇\苯酚\水脱保护,将多肽从MBHA树脂上切割下来。结果:粗肽的收率为62%,经RP-HPLC纯化,即可获得纯度在98%以上的目标肽。经MALDI—MS质谱分析其分子量与理论值一致。结论:此工艺操作简单,便于推广,适合大规模生产。     

人脑利钠肽的Fmoc固相合成

探讨生物活性肽人脑利钠肽(BNP)的固相合成工艺,并为工业化合成提供理论依据.本文以二氯三甲基树脂(以下简称为二氯树脂)为载体,采用9-芴甲氧羰基(Fmoc)保护的氨基酸,以1-氧-3-双二甲胺羰基苯骈三氮唑四氟化硼盐(TBTU)/1-羟基苯并三氮唑(HOBT)/二异丙基乙胺(DIEA)缩合,以碘作为环化试剂,用切割试剂将BNP粗品从树脂上切割下来.通过MALDI-MS质谱仪检测,所合成环肽的分子量与理论分子量一致,使用RP-HPLC液相色谱仪对合成的环肽进行纯化,得到的BNP纯度达到97%以上.本合成工艺具有快捷、简便、高效的特点,适合于大批量的生产目的肽.     

杂环肽FIK的Fmoc固相合成法研究

为研究二硫键成环的杂环肽FIK的合成工艺, 以Fmoc氨基酸为原料, 采用固相合成法, 经TBTU/HOBT/DIEA复合缩合剂催化合成直链肽, 再经I2氧化肽链上两个半胱氨酸的巯基生成分子内二硫键而得到目标环肽, 将其用切割试剂切割脱离树脂得到粗产品, MALDI-MS和RP-HPLC进行鉴定, 分析和纯化。产率可以达到18%, 纯化后纯度达97%以上, 经MALDI-MS和Ellman试剂检测确定为目标肽。该合成法高效, 简便, 快速, 目标肽收到较理想产率, 适合大批量生产。     

杂环肽FIK的Fmoc固相合成法研究

为研究二硫键成环的杂环肽FIK的合成工艺, 以Fmoc氨基酸为原料, 采用固相合成法, 经TBTU/HOBT/DIEA复合缩合剂催化合成直链肽, 再经I2氧化肽链上两个半胱氨酸的巯基生成分子内二硫键而得到目标环肽, 将其用切割试剂切割脱离树脂得到粗产品, MALDI-MS和RP-HPLC进行鉴定, 分析和纯化。产率可以达到18%, 纯化后纯度达97%以上, 经MALDI-MS和Ellman试剂检测确定为目标肽。该合成法高效, 简便, 快速, 目标肽收到较理想产率, 适合大批量生产。     

两种树脂对固相法合成丙型肝炎疫苗多肽的影响

本实验采用固相法合成丙型肝炎疫苗多肽,比较Wang树脂和二氯三苯甲基树脂（以下简称二氯树脂）作为载体时的连接率以及目标肽的产率和纯度。通过质谱鉴定分析所得目标多肽的分子量,用茚三酮法对两种树脂与氨基酸的连接率进行比较,用反相高效液相色谱（RP-HPLC）进行粗肽的分析和纯化。结果显示以二氯树脂作载体,第一个氨基酸的连接率,以及目标肽的纯度和产率都要明显高于Wang树脂,因此二氯树脂作载体合成丙型肝炎疫苗多肽更适合大规模工业化生产的要求。     

虎纹捕鸟蛛毒素-Ⅰ的化学合成及其结构与生理活性分析

应用芴甲氧羰基(Fmoc)固相方法化学合成了虎纹捕鸟蛛毒素-I,合成采用Fmoc氨基酸五氟苯酯,各侧链保护措施如下:羧基和酚羟基用叔丁基(tBu)保护,赖氨酸和组氨酸侧链用叔丁氧羰基(Boc)保护,精氨酸胍基用4-甲氧-2,3,6-三甲基苯磺酰基(Mtr)保护,半胱氨酸巯基用三苯甲基(Trt)保护.固相载体为需活化的乙二胺-聚乙二醇-聚苯乙烯(EDA-PEG-PS)树脂.合成产物(肽树脂)用苯甲硫醚-乙二硫醇-苯甲醚-三氟乙酸系统一步裂解以及去除侧链保护基,然后用二硫苏糖醇(DTT)还原,再通过氧化型和还原型谷胱甘肽系统促使其形成正确的二硫键配对.对HPLC分离纯化得到的合成产物,进行了氨基酸组成、肽谱、氨基酸顺序以及一维核磁共振谱分析.分析结果表明合成毒素具有与天然毒素相同的化学结构和空间构象.生理实验结果表明,其生物学活性与天然Huwentoxin-I也相同.     

α- 促黑激素的固相合成研究

目的:探索α-促黑激素的合成工艺。方法:采用多肽固相合成法制备α-促黑激素。以Rink amide-MBHA树脂为载体、使用Fmoc保护策略、TBTU、HOBt、DIEA为缩合剂体系,最后用TFA、苯甲硫醚、水、苯酚、乙二硫醇混合液将多肽从树脂上切割下来。结果:合成后的目标多肽产率达64.9%,经过RP-HPLC纯化纯度可达98%,质谱鉴定显示纯化产物与目标多肽理论相对分子质量一致。结论:该方法操作方便,反应结果稳定,为固相合成生产α-促黑激素提供了一种可行的工艺方案。     

Investigation of the mechanism of temperature sensitivity of the electroreceptors of ampullae of lorenzini

In experiments on Black Sea skates (Raja clavata), the potential of the receptor epithelium of the ampullae of Lorenzini and spike activity of single nerve fibers connected to them were investigated during electrical and temperature stimulation. Usually the potential within the canal was between 0 and –2 mV, and the input resistance of the ampulla 250–400 k. Heating of the region of the receptor epithelium was accompanied by a negative wave of potential, an increase in input resistance, and inhibition of spike activity. With worsening of the animal's condition the transepithelial potential became positive (up to +10 mV) but the input resistance of the ampulla during stimulation with a positive current was nonlinear in some cases: a regenerative spike of positive polarity appeared in the channel. During heating, the spike response was sometimes reversed in sign. It is suggested that fluctuations of the transepithelial potential and spike responses to temperature stimulation reflect changes in the potential difference on the basal membrane of the receptor cells, which is described by a relationship of the Nernst's or Goldman's equation type.I. P. Pavlov Institute of Physiology, Academy of Sciences of the USSR, Leningrad. I. M. Sechenov, Institute of Evolutionary Physiology and Biochemistry, Academy of Sciences of the USSR, Leningrad. Pacific Institute of Oceanology, Far Eastern Scientific Center, Academy of Sciences of the USSR, Vladivostok. Translated from Neirofiziologiya, Vol. 12, No. 1, pp. 67–74, January–February, 1980.     

Principles of organization and evolution of systems of regulation of functions

Evolution of living organisms is closely connected with evolution of structure of the system of regulations and its mechanisms. The functional ground of regulations is chemical signalization. As early as in unicellular organisms there is a set of signal mechanisms providing their life activity and orientation in space and time. Subsequent evolution of ways of chemical signalization followed the way of development of delivery pathways of chemical signal and development of mechanisms of its regulation. The mechanism of chemical regulation of the signal interaction is discussed by the example of the specialized system of transduction of signal from neuron to neuron, of effect of hormone on the epithelial cell and modulation of this effect. These mechanisms are considered as the most important ways of the fine and precise adaptation of chemical signalization underlying functioning of physiological systems and organs of the living organism