第一部份：TM-1-1磷酸酯衍生物之製備 第二部份：N-烷基苯-2-萘醯胺化合物之製備與其抗黃嘌呤氧化酶抑制之研究 第三部份：改良型鈀碳催化氫解反應於酚性生物鹼去酚基反應之研究

Part 1 Preparation of TM-1-1 phosphate ester derivatives Part 2 Preparation and anti-xanthine oxidase activity evaluation of N-arylalkyl-2-naphthamides Part 3 Removal of the phenolic groups in phenolic alkaloids by a modified Pd/C-catalyzed hydrogenolysis method

林敬婷(Chin-Ting Lin)

指導教授 ： 李水盛

國立臺灣大學/醫學院/藥學研究所/博士(2016年)

https://doi.org/10.6342/NTU201600540

摘要

第一部分 TM-1-1磷酸酯衍生物之製備
缺血性心臟病為世界主要致死原因之一，過去本實驗室於阿朴芬生物鹼之氮原子上導入乙氧烯愈創木酚基團，製備出具保護缺血再灌流造成之心臟傷害作用之化合物TM-1與其活性代謝物TM-1-1。然TM-1與TM-1-1於動物急毒性實驗中因會造成震顫之中樞神經副作用而使後續研究停擺。
本研究以富含於台灣雅楠之laurolitsine，經：氮甲醯化、酚基苄基化、去甲醯基、還原性氮上烷基化、與催化性氫化反應，大量製備TM-1-1且總產率可達71%。 TM-1-1經酚基磷酸化與去苄基反應製備其雙磷酸酯衍生物TM-1-1 DP。N-Formyllaurolitsine經：選擇性酚基甲基化、去甲醯基、還原性氮上烷基化、磷酸化、與去苄基反應製備其單磷酸酯衍生物TM-1-1 MPa/b。
TM-1-1 DP較TM-1-1具較佳之水溶解度，經大鼠模式證明具較佳之心臟保護作用，且於60倍有效劑量下無觀察到震顫之副作用。TM-1-1 DP與TM-1-1 MPa/b於大鼠代謝研究中可知雙磷酸酯化合物無法穿過血腦障壁，於活體內可逐漸代謝為單磷酸酯代謝物與TM-1-1，而2號位置磷酸酯較9號位置易降解，且單磷酸酯化合物會增加腦內分佈。

第二部分 N-烷基苯-2-萘醯胺化合物之製備與其抗黃嘌呤氧化酶抑制之研究
黃嘌呤氧化酶近年來為高尿酸血症或心血管代謝相關疾病之重要研究標的，過去本實驗室發現植物頭花香苦草所含1,4-benzodioxane型neolignans類成份，具有良好之黃嘌呤氧化酶抑制活性。以活性化合物hyprohombin C進行電腦分子模擬分析其與黃嘌呤氧化酶之交互作用，並依此模擬結果計算出具N-arylalkyl-2-naphthamides架構之化合物與黃嘌呤氧化酶有良好之結合潛力。
本研究以此系列化合物為目標化合物以驗證其實際黃嘌呤氧化酶抑制活性。分別製備重要中間產物6,7-dimethoxy-2-naphtolic acid（IX）與3,4-dimethoxyphenyl-alkylamines化合物（Xa-e），再經由醯胺偶合反應與去甲基反應而得一系列之N-arylalkyl-2-naphthamides化合物（VIIa-e）。化合物VIIa-e和allopurinol (IC50 26.6 μM)相比皆具有中到高之體外抑制黃嘌呤氧化酶之活性且其趨勢可驗證電腦分子模擬之結果，其中以化合物VIIc（IC50 21.6 μM）與VIId（IC50 22.6 μM）具有最佳之抑制活性。以化合物VIIc進行高尿酸血症小鼠動物實驗，證實其具有降低血中尿酸濃度之作用。

第三部分 改良型鈀碳催化氫解反應於酚性生物鹼去酚基反應之研究
本研究將經由酚基轉換為1-phenyl-1H-tetrazol-5-yl ether衍生物後，以鈀碳催化氫解反應以移除酚性羥基之方法，藉由鎂粉或乙酸銨之添加進行方法改良。以五種常見之異喹：phenanthrene、aporphine、pavine、protoberberine、與1-benzyltetrahydroisoquinoline架構之生物鹼為反應物進行討論。結果顯示，與單純鈀碳催化氫解反應相比，鎂粉或乙酸銨之添加可提升其反應完全性、反應速率與產率、減少催化劑鈀碳之用量，因此有利於應用在量化去酚性羥基生物鹼之製備，有助於生物鹼結構與活性關係之探討時，修飾化合物之製備。

關鍵字

TM-1-1 ； 磷酸酯衍生物 ； N-烷基苯-2-?醯胺 ； 黃嘌呤氧化?抑制劑 ； 催化氫解反應 ； 酚性生物鹼 ； 去酚基反應

並列摘要

Part 1. Preparation of TM-1-1 phosphate ester derivatives
Ischemic heart disease remains as the worldwide leading cause of death. Our lab has prepared the aporphine derivatives, TM-1 and its active metabolite TM-1-1, with incorporation of O-ethylene guaiacol moiety to the nitrogen atom. TM-1 exhibited good cardioprotective effect on ischemia/reperfusion animal models. However, the further study of TM-1 and TM-1-1 was pended due to CNS tremor side effect in the acute tocxicity animal study.
In this study, a larger scale preparation of TM-1-1 was accomplished with a 71% total yield from laurolitsine, which presented abundantly in Phoebe formosana, via N-formylation, O-benzylation, de-formylation, reductiove N-alkylation, and catalytic hydrogenolysis reaction. TM-1-1 DP, a diphosphate ester of TM-1-1, was prepared from TM-1-1 via O-phosphorylation and de-benzylation. TM-1-1 MPa/b, monophosphate ester of TM-1-1, were prepared from N-formyllaurolitsine via selective O-methylation, de-formylation, reductive N-alkylation, O-phosphorylation, and de-benzylation.
TM-1-1 DP has better water solubility than TM-1-1. It shows improved cardioprotective effect in vivo animal model and no tremor effect was observed at a dose 60 times as large as the effect dose. The result of rat metabolism study demonstrated that the diphosphate ester compound would not pass BBB and would metabolize to mono-dephosphorylated metabolites and TM-1-1. 9-phosphate ester would be more stable than 2-phosphate ester and the mono-phosphate ester compounds increase the distribution of brain.

Part 2. Preparation and anti-xanthine oxidase activity evaluation of N-arylalkyl-2-naphthamides
In the recent years, xanthine oxidase has been recognized as an important target for hyperuricemia or cardiovascular and metabolic disorders. Our lab has found that Hyptis rhomboides contains 1,4-benzodioxane type neolignans, which possess potent anti-xanthine oxidase activities. Computer assisted docking study was performed to analyze the interactions of potent inhibitor hyprohombin C with xanthine oxidase. Further virtual docking predicted a new series compounds with N-arylalkyl-2-naphthamides structure to have binding potential with xanthine oxidase.
In this study, N-arylalkyl-2-naphthamides were prepared to verify their anti-xanthine oxidase activity. The key intermediates, 6,7-dimethoxy-2-naphtolic acid (IX) and 3,4-dimethoxyphenyl-alkylamines (Xa-e), were synthesized individually. The target compounds, N-arylalkyl-2-naphthamides (VIIa-e) were further prepared via amide coupling and demethylation reaction. VIIa-e exhibited medium to high inhibitiory potency compared with allopurinol (IC50 26.6 μM) in in vitro xanthine oxidase assay and the result was corroborated the results from molecular docking study. The VIIc (IC50 21.6 μM) and VIId (IC50 22.6 μM) showed the best inhibitory potency. The in vivo assay on hyperuricemia mice model demonstrated that VIIc possessed the effect in reducing blood uric acid level.

Part 3. Removal of the phenolic groups in phenolic alkaloids by a modified Pd/C-catalyzed hydrogenolysis method
A modified palladium on carbon (Pd/C) catalyzed hydrogenolysis method for removal of the phenolic-OH group in phenolic alkaloids as the 1-phenyl-1H-tetrazol-5-yl derivative by the addition of magnesium metal or ammonium acetate in acetic acid is described in this study. Five different types of isoquinoline alkaloids, ie., phenanthrene alkaloids, aporphine, pavine, protoberberine, and 1-benzyltetrahydroisoquinoline, were used as reactants. The results indicate that the addition of either magnesium metal or ammonium acetate has the advantage of decreasing the amount of Pd/C and the accelerating reaction rate over the simple Pd/C-catalyzed hydrogenolysis, thus it is practical for larger-scale preparation of de-phenolated alkaloids for pharmacological study.

並列關鍵字

TM-1-1 ； phosphate ester derivatives ； N-arylalkyl-2-naphthamides ； xanthine oxidase inhibitors ； hydrogenolysis ； Removal of the phenolic groups ； phenolic alkaloids

參考文獻

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