多肽研究XX:丙型肝炎病毒(HCV)合成多肽的抗原性及线性抗体谱

依据蛋白质的亲水性、可亲性、柔韧性、抗原性、电荷分布及HPLC滞留系数等六种性质,使用“Goldkey”软件系统给出了HCV-BK各蛋白抗原决定簇预测曲线,共设计、合成15个多肽片段:P1(475～495),P3(449～468),P4(658～663),P5(645～663),P6(484～489),P7(475～489),P15(655～662),P16(230～237),P17(225～237),P18(1220～1240),P19(1694～1735),P24(1230～1240),P25(1482～1493),P26(384～389)和P27(2355～2389)。发现NS1区%和NS4区P19有很强的抗原性。用其检测PT-HC,阳性率分别为60%和63%。     

抗丙型肝炎病毒药物研究进展

丙型肝炎病毒(HCV)是导致慢性肝病和肝细胞癌的主要原因,越来越多的人感染HCV已成为严重的社会和公共卫生问题。由于HCV具有很强的变异性,目前尚无有效的疫苗。虽然新上市的telaprevir和boceprevir联合pegIFN(聚乙二醇-干扰素)和ribavirin治疗方法具有较强的抗HCV能力,可以有效的治疗部分HCV患者,但是仍具有较大的副作用和较差的药物耐受性,而二代直接抗病毒药物和宿主靶向药物的发现为治愈HCV患者提供了新的方向。     

检测丙型肝炎病毒抗体临床意义的评估

丙型肝炎病毒(HCV)分子克隆的成功和HCV抗体(抗-HCV)检测方法的建立,大大推进了全球的丙型肝炎(HC)研究和防治工作。为了帮助临床医师和流行病学医师进一步了解HC,本文对检测抗-HCV临床意义的评估综述如下。1 抗-HCV检测方法的建立七十年代初,在研究输血后肝炎时,证实了血传性非甲非乙型肝炎(PT-NANBH)的存在,但未发现其病原。自1982年,美国加利福尼亚州的Chiron公司开始对该病原进行研究:从已知含有高浓度感染物的黑猩猩血浆中,通过超速离心而获得了感染颗粒,提取其核酸,反转录为cDNA,通过多聚酶链反应(PCR)的方法使cDNA大量扩增,插入λgt_(11),建立基因文库,用已知PT-NA-     

丙型肝炎病毒与扁平苔藓相关性研究

为了确定丙型肝炎病毒 ( hepatitis C virus,HCV)与扁平苔藓 ( lichen planus,L P)之间是否存在某种联系以及丙型肝炎抗原 ( hepatitis C antigen,HCAg)阳性表达与 L P患者性别的关系 ,应用 s ABC免疫组织化学染色法对 3 1例 L P患者的病理切片进行了分析。结果显示 ,HCV在 LP的发病机理中的作用不可忽视 ;L P患者中 HCAg的表达虽然在不同的性别中表现形式有所不同 ,但无统计学意义。     

丙型肝炎病毒抑制剂研究进展

丙型肝炎病毒(hepatitis C virus, HCV)感染是全球性的公共卫生问题之一,全世界有1.3亿至1.5亿人长期感染,其中四分之一的患者会产生肝硬化、肝细胞癌甚至肝功能衰竭等并发症。完全清除病毒是研究者不断进行抗丙肝新药物研发的目标与动力。本综述精选近几年具代表性的研究实例,从药物化学的视角总结了抗丙肝小分子抑制剂的前沿进展。     

多肽研究——Ⅸ.乙型肝炎病毒表面抗原(HBsAg)Pre-S区域肽段的合成和抗原特异性测定

采用溶液法和固相法,合成了HBsAg Pre-S蛋白区域七个六肽片段。将其与载体蛋白结合。用多克隆抗HBsAg和单克隆抗“α”测定了肽—蛋白结合物的抗原特异性,结果表明P_1和P_5有较高的抗原特异性。     

多肽研究——Ⅸ.乙型肝炎病毒表面抗原(HBsAg)Pre-S区域肽段的合成和抗原特异性测定

采用溶液法和固相法,合成了HBsAg Pre-S蛋白区域七个六肽片段。将其与载体蛋白结合。用多克隆抗HBsAg和单克隆抗“α”测定了肽—蛋白结合物的抗原特异性,结果表明P₁和P₅有较高的抗原特异性。     

乙型和丙型肝炎病毒与再障的相关性

目的 探讨乙型和丙型肝炎病毒与再障的关系。方法 采用分子生物学技术方法,检测了26例首发再生障碍性贫血和65例随机正常人血清中的乙肝炎病毒和丙型肝炎病毒的阳性率。结果 再障组中乙型肝炎病毒（HBV）和丙型肝炎病毒（HCV）阳性率分别为26．89％和46．15％,而在正常人对照组中HBV和HCV的阳性率则分别为10．64％和1．54％,两组比较,HCV在再障组中明显升高,具有显著性差异（P＜0．01）,HBV未见强相关。结论 HCV感染可能是造成再生障碍贫血的主要原因。     

酪-D-丙-甘-苯丙-D-亮(DADL)五肽的固相合成及对免疫功能的影响

用固相多肽合成法合成了脑啡肽衍生物DADL,一种δ受体选择性激动剂。实验观察发现DADL能直接促进小鼠脾淋巴细胞增殖,并有一定浓度依赖关系,与ConA和LPS无明显协同效应,此外DADL(l.0mg·kg^-1X6d)在体内能促进TNF-α的产生。体外高浓度时(10^-6～5Xl0^-6mol·L^-1)能促成LPS诱导的TNF-a的表达。上述结果表明,DADL与脑啡肽类似,有一定免疫调节作用。     

丙型肝炎病毒NS5A抑制剂——达卡他韦

达卡他韦(Daklinza)是由美国百时美-施贵宝公司(BMS)研制的口服丙型肝炎病毒(HCV)NS5A抑制剂。继2014年欧盟批准其与联合其他抗病毒药用于基因型1-4慢性丙型肝炎成人患者的治疗后,2015年7月美国FDA批准其与索非布韦[sofosbuvir]联合用于慢性HCV基因型3感染的治疗。达卡他韦是第一个无需同时给予干扰素或利巴韦林即可有效治疗基因型3 HCV感染的药物,为该类患者提供了一个新选择,包括那些不能耐受利巴韦林患者。笔者就达卡他韦的基本信息、作用机制、药代动力学、药物相互作用、临床试验及应用等研发动态作一概述,以期能为医院临床用药及药物研究开发提供参考。     

STUDIES ON PEPTIDES

The tetradecapeptide amide corresponding to the entire amino acid sequence of a wasp venom (polistes mastoparan), isolated from Polistes jadwagae, was synthesized using the trifluoroacetic acid-thioanisole deprotecting procedure.     

根结线虫数值分类方法的筛选研究

本研究对Melo■属60个OTUS和18项分类性状分别进行Q分类和R分类,结果表明;Q分类以在切比雪夫距离上用中间距离法进行聚类效果最佳,能概括和直观地表达根结线虫的表相关系;R分类在马氏距离上用中间距离法聚类能很好地表达数量性状之间、质量性状之间以及数量性状与质量性状之间的表相关系。Q分类结果还表明,60个OTUS的单位距离(种间)不存在属的差异,应取消H(?)属,原H(?)属中的种均应归属于Mel(?)属,M.(?)(?)并非是M.(?)的亚种,应提升为独立的一个种。     

Small-molecule inhibitors of hepatitis C virus (HCV) non-structural protein 5A (NS5A): a patent review (2010-2015)

Introduction: Non-structural 5A (NS5A) protein has achieved a considerable attention as an attractive target for the treatment of hepatitis C (HCV). A number of novel NS5A inhibitors have been reported to date. Several drugs having favorable ADME properties and mild side effects were launched into the pharmaceutical market. For instance, daclatasvir was launched in 2014, elbasvir is currently undergoing registration, ledipasvir was launched in 2014 as a fixed-dose combination with sofosbuvir (NS5B inhibitor).

Areas covered: Thomson integrity database and SciFinder database were used as a valuable source to collect the patents on small-molecule NS5A inhibitors. All the structures were ranked by the date of priority. Patent holder and antiviral activity for each scaffold claimed were summarized and presented in a convenient manner. A particular focus was placed on the best-in-class bis-pyrrolidine-containing NS5A inhibitors.

Expert opinion: Several first generation NS5A inhibitors have recently progressed into advanced clinical trials and showed superior efficacy in reducing viral load in infected subjects. Therapy schemes of using these agents in combination with other established antiviral drugs with complementary mechanisms of action can address the emergence of resistance and poor therapeutic outcome frequently attributed to antiviral drugs.     

美沙酮维持治疗者合丙肝患者的HCV—RNA定量与ALT检测结果分析

目的：了解美沙酮维持治疗者合并丙型病毒性肝炎者HCV—RNA定量与ALT的检测情况,以明确在该人群中丙肝病毒复制、传染程度及需要治疗人数比例。方法：采集53例美沙酮维持治疗者合并丙肝患者的血液样本进行HCV—RNA定量与ALT检测,并对检测结果进行分析。结果：美沙酮维持治疗者合并丙肝患者53例中HCV—RNA定量检测结果的异常率为77．36％（41／53）;ALT异常率73．58％（39／53）;HCV—RNA定量异常者并存ALT异常比例为87．80％（36／41）,HCV—RNA定量正常者ALT异常率为25．00％（3／12）,两者比较有统计学差异（X^2=18．838,P〈0．01）。结论：美沙酮维持治疗者合并丙肝患者当中丙肝病毒复制活跃、传染程度高及需要治疗人数比例高,增加了丙肝传播风险和社会负担等社会公共卫生问题,建议政府完善对该人群现存丙肝的防控措施和政策。     

STUDIES ON TRYPSIN INHIBITORS

The synthesis by fragment condensation of protected peptides corresponding to the amino acid sequences 15–35, 25–52 and 15–52 of porcine pancreatic secretory trypsin inhibitor II (Kazal type) is described. The Rudinger modification of the azide procedure was used in the fragment coupling steps. The tert-butyloxycarbonylheptapeptide hydrazide (sequence 22–28) was reacted with the heptapeptide methyl ester free base (sequence 29–35) and the resulting tert-butyloxycarbonyltetradecapeptide methyl ester after selective deprotection, coupled with the benzyloxycarbonylheptapeptide hydrazide (sequence 15–21) to give the protected peptide methyl ester corresponding to the 15–35 sequence which was then converted to the corresponding hydrazide. The synthesis of the 25–52 sequence was achieved by assembling the protected peptide hydrazide corresponding to the amino acid residues 25–35, with the C-terminal heptadecapeptide 36–52. The resulting protected octaeicosapeptide (sequence 25–52) was selectively deblocked with trifluoroacetic acid and acylated with the benzyloxycarbonyldecapeptide hydrazide 15–24 to give the desired octatriacontapeptide corresponding to sequence 15–52 of the inhibitor. An attempt to prepare the 15–52 sequence through the condensation of fragments corresponding to 15–35 and 36–52 sequences was unsuccessful. The identity and purity of the synthetized peptide derivatives were established by elemental analysis (in some cases), amino acid analysis, optical rotation, and thin-layer chromatography in two solvent systems. The final products were also evaluated, after partial deprotection with anhydrous hydrogen fluoride or aqueous 90% trifluoroacetic acid, by paper electrophoresis at different pH values.     

STUDIES ON PEPTIDES

The heptacosapeptide amide corresponding to the entire amino acid sequence of gastrin-releasing peptide (GRP) was synthesized by assembling seven peptide fragments followed by deprotection with a new reagent system, IM trifluoro-methanesulfonic acid-thioanisole in TFA. The deprotected peptide was purified by ion-exchange chromatography on CM-cellulose followed by partition chromatography on Sephadex G-25. The latter was found to remove effectively the Met (O) derivative of GRP. The highly purified synthetic GRP was active as synthetic bombesin on the molar basis. A new carboxyl-activating reagent, thiazoline-2-thione, was employed for preparation of the necessary fragments.     

194例海洛因依赖者乙、丙肝炎病毒感染情况

采用ELISA法检测验194例海洛因依赖者HCV ,HBsAg 的感染情况 ,结果显示海洛因依赖者HBsAg,HCV的感染率分别是23.2 %和33.5 % ,显著高于对照组。尤其是iv滥用者感染率更高。本文认为HBsAg 和HCV的高感染率与iv海洛因有明显关系。