2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/37557AIDS is a pandemic which portends an increasing toll in human suffering and draining of national resources. Current approaches for the treatment of AIDS using single agents are plaqued by the development of tolerance. Combination therapies employing multiple components, each directed against different viral enzymes, may potentially provide an effective means of countering such tolerance. The HIV reverse transcriptase and protease enzymes are two targets which currently provide the basis for most AIDS therapy. To augment these approaches, inhibitors directed at new enzyme targets are needed, with the HIV integrase enzyme being one such important enzyme. Its attractiveness is attributed to the absolute requirment for its participation in HIV replication. The enzyme functions in a two-step manner by initially removing a dinucleotide unit from the 3??-ends of the viral DNA. The 3??-processed strands are then transferred from the cytoplasm to the nucleus where they are introduced into the host DNA following 5-base pair offset cleavages of opposing host strands. Propolis, a honey bee hive product, exhibits a broad spectrum of activities including antibiotic, antiviral, anti-inflammatory and tumor growth inhibition; some of the observed biological activities may be due to caffeic acid (cinnamic acid) esters that are present in propolis. In the present study we synthesized five caffeic acid esters, namely 3-(3,4-dihydroxypheyl)acrylic acid phenethyl ester (CAPE), 3-(3,4-dimethoxyphenyl)acrylic acid phenthyl ester (PEDMC), 3-(3,4-dihydroxyphenyl)acrylic methyl ester (MC), 3-(3,4-dihydroxyphenyl) acrylic acid ethyl ester (EC), 3-(4-bromophenyl)acrylic acid phenethyl ester (BrCAPE), and four structure-like compounds, namely 3-(3,4-dimethoxyphenyl)acrylic acid ethyl ester, 3-(4-methoxyphenyl)acrylic acid ethyl ester, 2,3-dibromo-3-(3,4-dimethoxyphenyl)propoinic acid ethyl ester, 2,3-dibromo-3-(3,4-dimethoxyphenyl)propoinic acid ethyl ester and tested them against the PBMC cell infected by M-tropic (JRCSF), T-tropic (NL-4-3) and Dual tropic (89.6) of HIV-1 isolates. CAPE and MC shown significant inhibition on the growth of HIV-replication in PBMC cell, but PEDMC shown less inhibition effect than CAPE and MC. Also, the effect of these agents on the growth of normal human buccal mucosal fibroblast (BF), oral submucosus fibroblast (OSF), neck metastasis of Gingiva carcinoma (GNM), tongue squamous cell carcinoma (TSCCa) was studied by MTT assay. Treatment with 25-200 mM CAPE and 50-400 mM EC shown significant cytotoxicity on OSF, GNM, TSCCa cells, but not on BF cell. The results suggested that CAPE-like compounds may be good chemotherapy agents against HIV infection and oral cancer.愛滋病（AIDS）或稱後天免疫缺乏症候群（Acquired Immunodeficiency Syndrome）之致病原HIV是一種RNA病毒，喜好侵犯人體血球中的T淋巴球及巨噬細胞，因此造成T淋巴球數目減少及細胞免疫能力的降低，從而各種伺機性感染趁機發生。在1997年後，使用強而有力三合一「雞尾酒式合併療法」做早期治療（Hit early, hit hard）已成主流。這種轉變歸因於對愛滋病毒的進一步了解與更多抗病毒藥物的發明。合併療法的主要考量在於愛滋病毒為一種易生突變的病毒，突變之後病毒可產生抗藥性，若使用單一藥物，很快便失去療效，又因有些藥物間有交叉抗藥性，使得他種同類藥也喪失功效。使用合併療法後，由於病毒必須產生更多的突變來產生抗藥性，基本上這樣的變種病毒無法在自然界繁衍，因此能長期控制病情，但因體內受愛滋病毒感染的巨噬細胞等長壽命細胞可為病毒的庇護所，所以合併療法一定要持之以恆，才能將潛藏的病毒一併消滅。 合併療法有各種不同組合，原則上以兩種反轉錄酵素抑制劑配合一種蛋白酵素抑制劑構成三合一療法，從不同方位攻擊病毒的複製與繁衍，使體內病毒量降低至最少的程度。目前臨床上採用何大一博士發明的合併療法來治療愛滋病病人取得重大的成效，但由於仍有病毒存有抗藥性之問題，因此開發新藥針對目前以反轉錄酵素及蛋白質酵素為主的其他抑制性藥物極為迫切。為了加強這些治療策略，針對其他病毒酵素的抑制物就成了重要的目標。在HIV本身的酵素當中與反轉錄酵素及蛋白質酵素同樣由Pol基因負責製造的還有嵌合脢。嵌合脢的存在對於HIV的複製是必需的，目前已可利用in vitro assay來測試各種抑制物抑制酵素活性之效果。 蜂巢臘膠(Propolis)為蜜蜂蜂窩之產物，近年來普遍被認為具有一系列之活性，包括抗菌、抗病毒、抗發炎及腫瘤抑制等。而其中的CAPE(Caffeic acid phenethyl ester)被發現為這些活性之主要成份。CAPE類的一系列構造類似物最近有研究顯示與目前已知的HIV integrase inhibitor結構非常相似。本計劃因此針對CAPE之類似衍化物4-溴肉桂酸苯乙酯之細胞毒性及ex vitro(virology)抑制HIV之複製效果做一完整之研究。以MTT assay分析9種不同化合物對於正常及癌細胞的細胞毒性，結果CAPE 和EC在100 mM能抑制約50 % OSF cells、GNM cells、TSCCa cells的生長，到200 mM時會使這些癌前細胞或癌細胞的生長降低到20%以下，對正常細胞無抑制生長的作用；含Br-基的化合物，除BrCAPE對GNM（齒齦癌細胞）有明顯細胞毒性外，2,3-dibromo-3-(3,4-dimethoxyphenyl)-propionic acid ethyl ester和2,3-dibromo-3-(4- methoxyphenyl)-propionic acid ethyl ester並無法有效抑制癌細胞生長，故推測caffeic acid系列的結構具有兩個芳香環或中間有脂類連接和含有羥基，在治療口腔癌部分有明顯效果，因此可深入探討作用機制。以CAPE、PEDMC、MC和BrCAPE作用於抑制HIV複製上，利用三株不同的HIV-1 isolates [M-tropic (JRCSF), T-tropic (NL-4-3) and Dual tropic (89.6) ]，結果顯示CAPE和MC對於HIV之複製有明顯抑制作用，於濃度10 mM至100 mM之間尤其明顯，但PEDMC和BrCAPE的抑制作用則沒有CAPE和MC明顯。這初步結果顯示CAPE類的化合物可能有抑制HIV複製之作用。本計劃因此進行更深入的研究、測試integrase的活性是受到該化合物之抑制，並明確了解其抑制目標及抑制HIV之機制。此計畫研究成果或可用來做為將來臨床治療愛滋病藥物開發之參考。HIV嵌合脢抑制物4 - 溴肉桂酸苯乙酯HIVintegrase inhibitor4-bromocinnamic acidProject