蟾酥水溶性抗肿瘤活性成分及其作用机制研究

Toad venom is China Pharmacopoeia varietieswith detoxification, relieve pain, the resuscitative effect.In recent years, it was widely used as antitumor drugs.Bufadienolides is generally considered as the main active ingredients.But widely used in clinic Bufo anti-tumor agents, for example, Chan′su injection and Cinobufacini injection, contain almost no bufadienolide compounds. Dramatically wd found after completely removal of the bufadienolides thewater extract of Chan′sustill keepexcerllent anti-tumor effect in vitro.The inhibition rateon lung cancer cell line was 96.4% at 1.0 mg/ml dose. And the acute toxicity test showed the water extract ofChan′su was far lower than the bufadienolides composition. So it is could be pridicted that toad venom water-soluble components may also is another main anti-tumor activity substance.This project adopts the methods of anti-tumor active tracking to separatethe water-soluble components of toad venom, investigate the in vitro antitumor activities of compounds from the water-soluble components and structure-activity relationshipand explore the anti-tumor mechanisms of active components on the molecular level.It is will be the first time to presentthe active anti-tumor components fromwater-soluble extract of toad venom and it is also the first time to clarify theiranti-tumor mechanisms in the world.This project will bethe more important scientific basis for the clinical safety of the drug originated from toad, and will be advantageous to improve quality control specification of correlative drugs. This project will also carry ona very positive impact on the development of new anti-tumor drugs.

蟾酥为药典品，具有解毒止痛，开窍醒神的功效。近年来作为抗肿瘤药物而广泛使用。目前普遍认为脂溶性的蟾蜍甾烯类成分为蟾酥抗肿瘤主要活性物质，但临床广泛使用的蟾蜍类抗肿瘤制剂蟾酥注射液和华蟾素注射液中基本不含有蟾蜍甾烯类成分，同时本课题组前期研究发现，完全去除脂溶性成分后所制备的水溶性蟾酥提取物仍具有很好的体外抗肿瘤活性，对A549肺癌细胞株，在1.0 mg/ml剂量下即可达到96.4%的抑制率，同时其急性毒性远低于蟾蜍甾烯类成分。故推测蟾酥水溶性成分亦可能是蟾酥抗肿瘤的主要药效物质。本课题采用活性追踪方式系统分离蟾酥水溶性成分，考察单体化合物体外抗肿瘤活性和构效关系，从分子水平探究活性成分作用机制。将首次明确蟾酥水溶性抗肿瘤活性成分，首次其作用机制，将对蟾蜍类药材及其制剂的临床安全用药提供更加科学的依据，对于更加合理地制定相关制剂质量控制方法、发现新型抗肿瘤药物等也都将产生非常积极的影响。

蟾酥为药典品，具有解毒止痛，开窍醒神的功效。近年来作为抗肿瘤药物而广泛使用。课题组前期研究发现，完全去除脂溶性成分后所制备的水溶性蟾酥提取物仍具有很好的体外抗肿瘤活性，因此采用体外活性追踪方式系统分离蟾酥水溶性成分，从中共分离得到38个化合物，鉴定了其中的29个，包括17个蟾毒色胺类化合物，3个有机酸类化合物和9个蟾毒配基类化合物。其中新化合物4个，新天然产物2个，蟾酥中首次分离得到的化合物2个。蟾毒色胺类化合物N-{5-acetyl-1-[2-(5-hydroxy-1H-indol-3-yl)ethyl]-1H-pyrrol-3-yl}acetamide显示了明显的体外抗肿瘤活性，为主要的蟾酥水溶性抗肿瘤活性成分之一。其它蟾毒色胺类化合物未显示明显的体外抗肿瘤活性。蟾酥水提物中含有痕量的蟾毒配基类成分，可能是蟾酥水提物中发挥抗肿瘤活性的主要成分。蟾毒色胺类化合物与蟾毒配基类合用并不能增强蟾毒配基类的细胞毒作用。本课题的研究成果对蟾酥及蟾蜍类药材及其制剂的药效物质阐明、质量标准建立及提升、蟾毒配基类成分生物合成途径的解析及相关产品的开发改造等提供了重要的科学依据，具有重要的科学及应用价值。