二烯丙基二硫化物是大蒜的活性成分之一,但其提取和合成产率较低。本研究拟通过结构修饰和活性研究，获得高产率和高抗肿瘤活性的化合物。基于前期工作，研究内容如下：1）在保留二烯丙基二硫化物的双硫键和对称式烯键的结构基础上对其进行改造，纯化、鉴定目标产物，实现产物结构多样化；2）建立马立克病毒鸡胚成纤维细胞模型，对二烯丙基二硫化物及其修饰产物进行药效、构效分析，筛选高活性产物，阐明其药效活性的关键结构域；3）以马立克氏病肿瘤细胞系MSB-1细胞为研究对象，通过流式细胞分析和Western-blot等方法，检测二烯丙基二硫化物及其修饰产物对MSB-1细胞的细胞凋亡及Caspase酶等相关凋亡蛋白表达的影响，探讨其抗病毒性肿瘤的作用机理。本项目的实施不仅为以二烯丙基二硫化物类活性成分为先导化合物开发新药提供理论依据，对病毒性肿瘤的深入研究和鸡马立克病毒病的防治也将起到推动作用。

Diallyl disulfide is one of the major compounds of garlic, but its efficiency of extraction and synthesis is lower. Our study plan to investigate the structure modification and activity of diallyl disulfide, and to obtain the compounds of high yield and high anti-tumor activity. Based on preliminary work, the contents are as follows: 1) we will make some alterations based on the disulfide bones and symmetric alkene bones, purify and identify the objective product, and obtain produts of diversified structure; 2) Establishing the cell model of Chick embryo fibroblast(CEF) infected with Marek′s disease herpesvirus( MDV), the pharmacodynamic and structure-activity relationship of diallyl disulfide and its modified products were analyzed, the high activity products were screened, and its key structure domains with drug effect were clarified. 3) In order to study the possible molecular mechanism of antitumor effect of diallyl disulfide compounds. Using the MDV MSB-1cells, the effect on cell apoptosis and expression of apoptosis protein of diallyl sulfide compounds were detected through western-blot method. The implementation of this project could not only provide theoretical foundation for developing new drugs using the active composition of diallyl sulfide as the lead compound, but also promote the deep study of viral oncology and the prevention and cure of MDV.

二烯丙基二硫化物是大蒜素中物化性质较为稳定的有效活性成分，已有大量研究报道了大蒜里面所含的二烯丙基二硫化物能够抑制多种肿瘤细胞的细胞增殖，并诱导细胞凋亡。本课题以鸡马立克氏病肿瘤细胞系MDCC-MSB-1细胞为研究对象，对二烯丙基二硫化物及其修饰产物诱导病毒性肿瘤细胞的凋亡机制进行了深入的探讨。 结果发现，通过MTT和流式细胞分析法对合成的DADS和1,2-二(2-甲基烯丙基)二硫醚和1-(4-甲基-2-戊烯基)-2-(4-苯基-2-丁烯基)二硫醚进行初步抗肿瘤活性检测，发现DADS和1,2-二(2-甲基烯丙基)二硫醚和1-(4-甲基-2-戊烯基)-2-(4-苯基-2-丁烯基)二硫醚能够浓度依赖性的抑制细胞增殖和诱导细胞凋亡；通过构效分析可知，亲脂性基团的引入可以增强烯丙基二硫化物类化合物的抗病毒性肿瘤能力；通过建立马立克病毒鸡胚成纤维细胞模型，在安全浓度范围内，检测二烯丙基二硫化物抗马立克氏病毒（MDV）能力，结果表明DADS可抑制MDV的扩增，且DADS在浓度较高的水平下能明显抑制MDV造成的机体器官损伤，减轻病症，但DADS最适治疗剂量、剂型仍需后续试验研究；通过CCK-8法定量和吖啶橙（AO）染色法定性分析发现DADS能剂量性降低MDCC-MSB-1细胞的增殖速度；流式细胞术检测出DADS能诱导MDCC-MSB-1细胞发生G2/M期细胞周期停滞，降低细胞的分裂速度；在线粒体介导的胱天蛋白酶依赖的凋亡途径中，发现凋亡诱导蛋白Caspase-9和Caspase-3的mRNA表达量随着DADS浓度的增加而增加。总之，DADS能够有效抑制MDCC-MSB-1细胞分裂并通过介导细胞内Caspase级联反应产生凋亡效应。上述结果表明，烯丙基二硫化物类化合物的研发对病毒性肿瘤防治具有重要的意义。