基于斑马鱼模型的深远海真菌中促血管生成活性成分的研究

Pathological angiogenesis is a hallmark of many diseases. It is of significant importance for the treatment of ischemic diseases (for example, coronary heart disease) to find the highly active and lowly toxic pro-angiogenic drug. Deep-sea fungi are an important source of bioactive natural products, because that the extreme living environment increases the likelihood of strains to metabolize structurally unique compounds with various activities. With the unique biological characters, zebrafish has become an important model animal for angiogenesis-associated diseases and drug screening research. In previous studies, the extract of an Indian Ocean fungus Y32 exhibited strong pro-angiogenic activity, and a series of active alkaloids were isolated from it. This project primarily aims to quickly track the bioactive constituents with pro-angiogenic effects of three active fungi from Indian Ocean under the guidance of the transgenic zebrafish model. Then, we will identify the structures of compounds and reveal their structure-activity relationship. The pro-angiogenic metabolites of deep-sea fungi will be found efficiently by closely integrating the chemistry and biological activity. This research would contribute to providing novel structures for lead compounds of treating the ischemic diseases and new approach to utilize the resource of deep-sea fungi.

血管生成涉及多种疾病，寻找高效低毒的促血管生成药物对于治疗冠心病等缺血性疾病的意义重大。深远海真菌由于特殊的生存环境，蕴藏结构新颖、活性多样的次级代谢产物，成为天然产物活性成分的重要来源。斑马鱼因其独特的生物学特征，成为血管生成相关疾病模型和药物筛选研究的重要模式动物。前期研究发现印度洋真菌Y32的发酵产物显示强烈的促血管生成活性,并从该菌株中分离得到了一系列的生物碱活性成分。本项目拟利用转基因斑马鱼促血管生成模型，结合各种现代色谱分离技术，从三株活性显著的印度洋真菌中定向、快速的追踪其活性成分，并鉴定化合物的结构，初步揭示其构效关系。课题将化学和生物活性紧密结合，高效快速的发现深远海真菌中具有促血管生成活性的代谢产物，有望为缺血性疾病药物研发提供新的先导化合物，为深远海真菌资源的利用提供新思路。

血管生成涉及多种疾病，促进血管新生实现血运重建也是冠心病等缺血性疾病的治疗关键。深远海真菌由于特殊的生存环境，蕴藏结构新颖、活性多样的次级代谢产物，成为天然产物活性成分的重要来源。本项目利用转基因斑马鱼促血管生成模型，筛选了3株活性显著的印度洋真菌Penicillium expansum、Aspergillus Austroafricanus、Acremonium sp.。结合各种现代色谱分离技术，对活性菌株进行了系统的化学成分研究，分离得到67个化合物：包括40个生物碱类、6个萜类、5个甾体类、4个神经酰胺类、2个氧杂蒽酮类、2个硫醚类、1个苯醚类化合物、以及7个其它芳香类化合物，其中新化合物7个，新天然产物1个。通过斑马鱼模型对分到的化合物进行促血管生成活性评价，其中，氧杂蒽酮-蒽醌杂二聚体JBIR-99、生物碱cottoquinazoline A、倍半萜类(S)-4-(2,6-dihydroxy-6-methylheptan-2-yl)-3-hydroxybenzoic acid表现了明显活性且对斑马鱼没有毒性；苯醚diorcinol和生物碱类化合物4-(3-hydroxyphenyl)-3-methoxyquinolin-2(1H)-one对斑马鱼有较强毒性，细胞毒活性研究发现两化合物对肺癌细胞A549均显示一定的生长抑制活性；两个苯甲酸类化合物4-hydroxyphenylacetic acid和phenyl acetic acid表现出一定的血管抑制活性。