基于贝壳杉烷型二萜骨架的新型乙酰肝素酶抑制剂的设计、合成及抗乳腺癌肿瘤转移活性评价

Breast cancer is one of the most common malignant tumors all over the world. Most breast cancer can be controlled by aggressive treatment. However, metastasis is the leading cause of death. During hematogenous metastasis, tumor cells activated the coagulation system to create conditions for metastasis. And heparanase (Hpa) is a key target. A kind of kaurane diterpenoid named ent-16β,17-dihydroxy-kauran-19-oic acid (DKA) was isolated from Siegesbeckia. Their inhibition effects on MBA-MD-231 were evaluated to assess the potentiality of anti-metastasis for the first time, and showed that it had good activity. In order to find a new type of Hpa inhibitor with more potential for development, we will design and synthesize new derivatives using this diterpenoid as a lead compound. The anti-metastasis mechanism of the diterpenoid and its derivatives will be revealed by the virtual screening of computer-aided drug design (CADD) using heparanase as target, combined with the evaluation of the inhibitation of Hpa in vitro and anti-metastasis activity in vivo and in vitro. We will also explore the structure-activity relationships of them. All these above will provide theoretical basis and technical support for the development of new small molecular as heparanase inhibitors and anti-breast cancer metastasis drugs.

乳腺癌是女性最常见的恶性肿瘤之一，大多数乳腺癌可以通过积极治疗得到控制。但是，无论初期诊断时发现还是复发时的转移都是其治疗失败的主要原因。研究表明肿瘤细胞会激活凝血系统为其转移创造条件，而乙酰肝素酶是此过程的关键酶，这为研究活血化瘀中药或单体抗肿瘤转移提供了理论依据。本课题通过文献调研及抗血栓活性介导的化学成分研究，从豨莶草中分离得到一个贝壳杉烷型二萜—DKA，并首次对该二萜进行了抗MBA-MD-231转移活性筛选，发现其具有显著的活性。为了发现更有开发潜力的新型Hpa抑制剂，本课题拟以DKA为先导化合物，设计合成系列结构新颖的衍生物。通过以Hpa为靶标的计算机辅助药物设计结合体外Hpa抑制活性评价和体内外抗肿瘤转移活性评价，筛选出活性更好的候选化合物，并从分子水平探讨其抗肿瘤转移的作用机制及构效关系，为开发新型小分子乙酰肝素酶抑制及抗乳腺癌肿瘤转移药物提供理论依和技术支撑。

乳腺癌是全世界女性最常见的恶性肿瘤之一,严重威胁着女性的健康。从临床应用广泛的传统中药中寻找疗效好、副作用少的抗乳腺癌肿瘤转移药物是一种十分可行的途径。豨莶为菊科（Asteraceae）豨莶属（Siegesbeckia）一年生草本植物，为我国传统中药。现代药理实验表明豨莶草提取物具有抗炎、抗过敏、抗血栓、抗肿瘤、抗组胺释放的活性以及其他活性。DKA系从豨莶草中分离得到的活性二萜类化合物，具有较好的抗血栓作用，本课题组前期研究提示 DKA 具有显著抗MBA-MD-231转移活性。本课题通过划痕实验、小鼠乳腺癌肺转移模型实验，与Control组相比，DKA（1 μM, 5 μM和25μM）均可显著抑制细胞迁移，DKA(2.5 mg/kg, 5 mg/kg和10 mg/kg)均可以显著抑制MDA-MB-231乳腺癌细胞肺转移，进一步验证了DKA的抗乳腺癌MBA-MD-231的转移活性；通过对DKA及两种衍生物药代动力学研究发现，DKA在体内能够快速吸收，把邻二醇羟基和羧基上的氢替换掉之后，生物利用度显著提升；通过缩酮、酯化、酰胺化、糖基化等反应得到DKA衍生物42个，通过MTT实验得到6个具有细胞毒活性的衍生物，Transwell小室实验筛选得到2个活性较好的衍生物。迄今已发表文章2篇，其中SCI论文1篇，中文核心期刊论文1篇，提交专利申请1项。