合成物NP-184和bp5250對血管新生拮抗作用與機制之探討

The inhibitory effects and mechanisms of novel compounds, NP-184 and bp5250 on angiogenesis

林冠廷(Kuan-Ting Lin)

指導教授 ： 黃德富

國立臺灣大學/醫學院/藥理學研究所/博士(2012年)

https://doi.org/10.6342/NTU.2012.10054

摘要

失調的血管新生促使了許多疾病的演進，血管生長不良會造成心肌梗塞、中風、神經性退化疾病等，而過分生長或是不正常的血管則會促進癌症、眼部退化疾病和其它發炎相關疾病，因此，調控血管新生的過程提供了一個治療這些疾病的方向。腫瘤引發之血管新生是一個多項步驟的過程，在過程中，癌細胞誘發現有血管生成新的血管貼附。血管內皮生長因子是一個強力的血管新生誘發分子，它能被癌細胞所分泌，血管內皮生長因子能誘發內皮細胞增生、移行、分化來形成新的血管網絡以供給癌細胞生長。由於有多種的細胞和各種不同的拮抗或誘發血管新生分子參與其中過程，這提供了我們許多藥物研發的標的。雖然有數種抗血管新生藥物已經批准而臨床使用於治療眼部退化疾病和癌症，但是由它們的臨床結果，副作用以及產生的抗藥性來看，目前抗血管新生的治療仍然存在一定的挑戰。因此，繼續探索及發展新的抗血管新生藥物仍有其必要性。
在本論文中，我們研究兩種系列化合物NP-184和bp5250，它們經由生物活性篩選以確定其抑制血管新生的潛力。NP-184和bp5250都能呈現濃度相關性地抑制人類臍靜脈內皮細胞的增生、移行和管狀生成，它們也濃度相關性的抑制了活體外大鼠主動脈環的管腔生成現象，在體內實驗的部分，NP-184和bp5250都能在Matrigel plug assay中依劑量相關性地抑制血管內皮生長因子所誘發的血管新生。在血管內皮生長因子所誘發的訊息傳遞途徑分析中，NP-184降低了ERK和AKT的磷酸化以及NF-kappaB移轉到細胞核。而bp5250則減少了ERK, p38, PI3K, AKT, FAK, Src, Rac1/Cdc42和Rho的活化，另外在缺氧情況下，bp5250抑制了HIF-1alfa和血管內皮生長因子mRNA的表現。考量這兩個化合物亦具有明顯的抗血小板活性，NP-184和bp5250具有發展成双重功能的抗腫瘤血管新生的潛力。在未來的工作展望上，除了對於作用機制的進一步探討外，還希望藉由它們與標的結構的循理設計和活性分析的協助，來發展更具有抑制血管新生活性的藥物。

關鍵字

血管新生 ； 血管內皮生長因子 ； 內皮細胞

並列摘要

Dysregulated angiogenesis contributes to various diseases progression. Insufficient blood vessels growth causes myocardial infarction, stroke, and neuro-degeneration disorders while overgrowth or abnormal blood vessels promote malignant tumor, retinopathy and inflammatory diseases. Therefore, modulating angiogenesis processes provide a therapeutic direction for disease treatment. Tumor angiogenesis is a multistep process by which tumor cell induces new capillaries from pre-existing vessels. Vascular endothelial growth factor (VEGF) is a potent angiogenesis inducer which can be released by tumor cells. VEGF induces endothelial cell proliferation, migration and differentiation to form new vasculature for supporting tumor growth. Owing to the multi-involvement on types of cells and varieties of pro- and anti-angiogenic molecules, it reveals many target sites for the drug development. Although several anti-angiogenic drugs have been approved for eye degeneration and cancer treatments, the clinical outcomes, side effects and resistance remain challenges in these therapies.
In this report, we presented two serial derivative compounds, NP-184 and bp5250, which were screened for the inhibitory effect on angiogenesis by bioactivity assays. Both NP-184 and bp5250 inhibit the VEGF-induced proliferation, migration and tube formation of human umbilical vein endothelial cells (HUVECs) in a concentration- dependent manner. They also suppress the capillary sprouting from rat aortic ring ex vivo. As in vivo assay, NP-184 and bp5250 both dose-dependently repress new vessels invasion to the plugs in the VEGF induced Matrigel assay. In the VEGF activated signaling level, NP-184 reduces the phosphorylation of ERK and AKT, and NF-kappaB translocation while bp5250 decreases the activation of ERK, p38, PI3K, AKT, FAK, Src, Rac1/cdc42 and Rho. In addition, bp5250 inhibits the expression of HIF-1alfa and VEGF mRNA expression. Along with their additional antiplatelet activity, both compounds may provide additional advantages in vivo for suppression of tumor-induced angiogenesis, and hypercoagulation-induced tumor metastasis. However, detailed molecular mechanisms still need to be further investigated. Besides, optimization of the lead compounds by rational design and functional assays for improvement of potency is also required.

並列關鍵字

Angiogenesis ； VEGF ； Endothelial cell

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