登革病毒的感染已成為全球關注的公共健康問題，也是台灣嚴重流行感染疾病之一。登革病毒感染的症狀表現從輕症的登革熱到潛在致命的登革出血熱和登革休克症候群。然而，目前並沒有有效抗病毒藥物可用於治療登革病毒感染。登革病毒的複製涉及病毒蛋白和細胞因子複雜的交互作用，例如在登革病毒生命週期的各個階段中活化多種細胞激酶是必需的。PI3K / AKT信號途徑在許多細胞過程中有著關鍵的調節作用，而第二型登革病毒可以觸發此路徑來保護受感染的細胞避免細胞走向早期的凋亡性細胞死亡。此外，在登革病毒感染細胞中發現GRP78表現被上調控並且觀察到細胞內病毒抗原和傳染性病毒顆粒大量增加。Celecoxib衍生物，AR-12，是一個PDK-1抑制劑可以抑制PI3K / AKT信號並調控GRP78的表現。最近研究指出AR-12能夠抑制幾種出血熱病毒的複製，但AR-12在治療登革病毒感染的功效，目前尚不得而知。根據我們的實驗數據顯示，在登革病毒感染的不同細胞株中，給予AR-12後可以顯著降低登革病毒複製以及感染性病毒顆粒的釋放。因此，我們進一步探討AR-12是否透過PI3K/ AKT信號途徑與GRP78調控登革病毒的複製。我們的研究結果發現，AR-12降低了AKT的活性和GRP78在登革病毒感染細胞中的表達。此外，我們利用登革病毒感染的小鼠模型，在新出生的小鼠腦部及腹腔同時注射給予登革病毒與AR-12。結果顯示AR-12可以有效減少在感染小鼠中登革病毒的複製。我們進一步利用感染後再給予藥物的實驗來測試AR-12是否能治療登革病毒感染後的小鼠。我們發現，病毒感染後再投與AR-12同樣能有效的抑制登革病毒感染的細胞和小鼠。根據以上實驗結果，我們認為AR-12對於登革病毒的治療具有其潛在功效，在未來可作為一個具潛力的抗病毒藥物。

Dengue virus (DENV) infection has become a public health issue of worldwide concern and is a serious health problem in Taiwan. DENV symptoms range from dengue fever to potentially fatal dengue hemorrhagic fever and dengue shock syndrome. Yet there are no approved effective antiviral drugs to treat DENV. The replication of DENV involves complex interaction between viral proteins and cellular factors, such as various cellular kinases whose activities are essential for various stages of the DENV life cycle. PI3K/AKT signaling pathway plays a critical regulatory role in modulating many cellular processes triggered by DENV-2 to protect infected cells from early apoptotic cell death. In addition, GRP78 is up-regulated in DENV infected cells co-incident with high level intracellular viral antigen production and infectious virus release. AR-12, a derivative of celecoxib is a PDK-1 inhibitor to abolish PI3K/AKT signaling and suppresses GRP78 levels. It has been recently reported that AR-12 is able to block the replication of several hemorrhagic fever viruses, but the efficacy of AR-12 in treating DENV infection is still unclear. Here, we evaluated DENV replication in different cell lines including A549, Huh-7, U937 and Meg-01 in the presence of AR-12. DENV replication is significantly reduced in AR-12 treated cells, as well as the infectious viral particles. Hence, we further investigated whether AR-12 regulates DENV replication through PI3K/AKT signaling pathway and GRP78. According to our results, AR-12 reduces the expression of pAKT and GRP78 in DENV infected cells. Furthermore, we used a DENV infection mice model, in which the newborn mice were administered with DENV and AR-12 via intracerebral and intraperitoneal injection. As from the results obtained AR-12 showed a potential reduction of DENV replication in mice. And a post-infection treatment experiment was used to determine the therapeutic effect of AR-12 on DENV replication. We found that AR-12 also exhibited effective inhibition in DENV infected cells and mice model. In conclusion, we uncover the potential efficacy of AR-12 as a novel drug for DENV.

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