华蟾素注射液是由中华大蟾蜍皮肤精制得到的主要用于肿瘤治疗的中药注射剂，其含有的多肽成分占注射液固形物50%以上，这些多肽的抗肿瘤、镇痛等作用引起关注。然而，华蟾素注射液中肽类分子的来源与结构、具有特定生物活性的单体分子及其作用机制等许多重要问题有待研究。我们在中华大蟾蜍皮肤转录组测定的基础上，对华蟾素注射液蛋白质组和多肽组进行了测定，结果显示大分子的肽类物质广泛存在于注射液中。70种小肽以及来自127种蛋白质前体的多肽存在于注射液中，其中的3个小肽分子对肝癌、乳腺癌细胞有显著的抑制作用。本项目立足于前期工作，开展华蟾素注射液中肽类物质的系统研究，明确华蟾素注射液中小肽分子的所有结构，结合抗肿瘤、镇痛和致敏等生物学活性测定，进而阐明具有特定生物学功能小肽分子对华蟾素注射液疗效的贡献和对不良反应的影响。项目实施将为中药注射剂中蛋白质/肽类分子的研究提供新的方法和思路，促进中药现代化和国际化。

Cinobufacini injection, an aqueous extract from the skin of toad Bufobufogargarizans Cantor, is mainly used to treat various tumorin China.The content of protein-like components accounted for more than 50% of injection’s solids weight. These componentsshowed equal antitumor activities on different tumor cellscompared with the injection and raised much attention by investigators. However, the origin and the primary structures of the protein-like components, as well as the structure characterizedand activity specific peptides and the mechanisms of their action need to be investigated. Proteome and peptidome of the skin of Bufobufogargarizans Cantor were determined based on the assembly of the toad skin transcriptome. Our results indicated that the peptides widely existed in the cinobufacini injection. As many as 70 peptides and 127 proteinprecursorswere identified in the cinobufacini injection. Three out of the 70 peptides showed obvious inhibitory activities on tested tumor cells. Based on our foundings, this project will carry out systematically investigation onthe protein-like components contained in the cinobufacini injection. To reveal the natural property of peptides and evaluate their contribution to the drug efficacy and the adverse reactions by biological activity assays, such as anti-tumor, analgesic and sensitization activities. The proposed project will provide new method and ideafor the investigation of protein/peptide molecules intraditional Chinese medicine(TCM)injections, promote the modernization and internationalization of TCM industry.