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Title: | 第一部分:新穎醯胺基蒽醌類化合物的合成及生物活性研究
第二部分:5-羰基-5,6,7,8-四氫異喹啉之合成研究 PartⅠ:Synthesis and Biological Activity Studies of Novel Amidoanthraquinones PartⅡ:Study on Synthesis of 5-Oxo-5,6,7,8-tetrahydroisoquinoline |
Authors: | Huei-Ling Liou 劉惠玲 |
Advisor: | 忻凌偉(Ling-Wei Hsin) |
Keyword: | 抗癌藥物,合成,蒽,醌,類化合物, anticancer,anthraquinones, |
Publication Year : | 2008 |
Degree: | 碩士 |
Abstract: | 第一部分:
以平面三環為骨架的蒽醌(anthraquinone)類化合物長久以來被視為重要的抗癌藥物,如早期的doxorubicin與後來的Mitoxantrone(MX)與Ametantrone(AT)等,過去,一系列醯胺基化合物已被合成,經由醯胺鍵連結各種胺基酸,其中以接有蛋氨酸支鏈的化合物WRC-213的活性最好,推測可能是蛋氨酸支鏈上的硫原子與體內金屬進行螯合作用,而具活性。因此本研究以WRC-213和Mitoxantrone為lead compounds,側鏈中心的氮原子仍舊與三環上的NH基團保持二個碳數的距離,並在側鏈的上的不同位置,接上結構與蛋氨酸相似的支鏈,合成出一系列的蒽醌類化合物,如:Aminoanthraquinone系列、化合物12-16與化合物17-29,並將這些化合物進行多種生物活性藥理測試,評估比較其結構與活性間之關係(Structure and Activity Relationship, SAR)。 根據多種生物活性測試的結果,我們得到以下結論:1. 側鏈為三胺基二乙烷的ATM 系列與M1, M2 化合物中,蛋胺酸無論接在側鏈任何一個氮上都對活性沒有助益,保持側鏈中心的氮原子裸露,活性反而較好。2. 藥理數據發現,整體而言,碳硫支鏈接在側鏈上C 位置時,活性表現最佳。3. 以WRC-213 為前導化合物所合成出來的一系列化合物中,活性很好的化合物其結構特徵為:a. 支鏈距離醯胺基上的C 位置約四個碳的距離。b. 蛋胺酸上的硫原子若替換成氧原子或是亞磺醯基等陰電性類似的基團或是接上拉電子基團,如羰基或苯硫基,也會有很好的活性。c. 若醯胺基上C 位置上的NH 有被保護起來,活性會較差,有趣的是化合物27 與26,其胺基有無保護的藥理活性都很好,所以需要進行更進一步的藥理測試。 測量化合物與DNA 的結合能力可再度證實,WRC-213 系列衍生物的作用機轉主要不是透過與DNA的結合達到抑癌的效果,另外,由Comet assay與MTT assay的結果可發現,化合物26 與29 對血癌細胞株的DNA 的傷害程度都不高,但29對粒線體細胞卻有良好的抑制活性;而對前列腺癌細胞株有良好活性的26,對粒線體細胞則無毒殺效果,因此,這兩個化合物的作用路徑需再做更深入的探討,由此可知,藉由MTT assay 與Comet assay 的藥理測試, 可提供一個篩選有潛力藥物的方法。 第二部份: 本實驗室先前致力於合成有關嗎啡ACNO環的類似衍生物,在化學合成部份已發展出一系列的合成路徑,此合成路徑中,具有光學活性的化合物5-羥基-5,6,7,8-四氫異喹啉是主要的中間體,但要得到此中間體,其前驅物5-羰基-5,6,7,8-四氫異喹啉(5-Oxo-5,6,7,8-tetrahydroisoquinoline)的製備便十分重要,參考相關文獻發現,此化合物的產率並不高,也常生成在第8位置取代的酮基副產物,之前本實驗室合成到此中間體需要四個步驟,總產率為43%,反應途中會用到激烈的強氧化劑,於是,為了提升此化合物的產率與採用較溫和的合成方法,我們參考了相關的兩篇文獻:K. C. Nicolau與P. Lardenois的合成策略。 K. C. Nicolau 的方法是使用溫和的氧化劑(o-iodoxybenzoic acid, IBX)直接將5,6,7,8-四氫異喹啉(5,6,7,8-tetraisoquinoline)進行苯環上的氧化,利用一步反應即可得到酮化合物,由於此反應以TLC 進行監測有其困難度,所以我們利用高效 能液相層析儀(High Performance Liquid Chromatography, HPLC)找出反應中產量最大的時間點,接著改變條件,找出最佳化的條件與最大產率,更利用液相層析串聯式質譜儀鑑定出反應中間形成的副產物,結果發現產率並不如文獻所提及之70%,而只有26%,所以我們轉而參考另一個年代較久遠的P. Lardenois 的合成策略。 最後,我們參考P. Lardenois 的合成方法,以HPLC 作為反應中途的監測器,確定每一步產物的生成,反應完進行蒸餾,純化產物,得到了此在合成上具有高度價值的前驅酮化合物,並達到第一步產率為78%,第二步產率為91%,兩步總產率為71%的高產率,予以量產(scale-up)後,得到了100 g 的產物,如此一來,有關後續嗎啡類似衍生物合成的總產率便能大大的提升。 PartI: Derivatives of compounds containing planer tricylic skeleton are important anticancer drugs, such as doxorubicin, Mitoxantrone and Ametantrone. The limiting clinical use of doxorubicin is due to its cardiotoxicity. However, A series of amidoanthraquinones connected with various amino acids were synthesized, such as WRC-213 has the best potency. It’s probably because the sulfur-containing side chain presents in it may chelate the metal ion in vivo, and thus exert the activity. Therefore, WRC-213 and Mitoxantrone serve as lead compounds in this study to connect with methionine related analogues on different positions of the side chain, such as Aminoanthraquinones, compound 12-16 and 17-29. These compounds have been assayed for their bioactivities to evaluate their structure and activity relationship. According to the results of the bioactivity assays, we can get the conclusions following: 1. Methionine that attached to the diethylenetriamine side chain is not beneficial for the activity. To keep the central nitrogen atom of the side chain naked could increase the activity. 2. As a whole, the methylsufanyl group attached to the Cd has the best potency. 3. WRC-213 analogues which possess good potency have some characteristics: a. Side chain keeps the Ca of the amido group for four carbons. b. If the sulfur atom of the methionine is replaced by a group with similar electronegativity, like oxygen or sulfinyl group or connect with electron withdrawing group would have good activity. c.c. If the NH on C of the amido group is protected, the activity would be poor. However, compound 26 and 27 both have good potencies, further pharmacological assay will go on process. From the data of the ability of compounds intercalating to DNA, it reconfirms that the major anticancer mechanism of WRC-213 analogues are not through binding to DNA. In addition, the results of Comet assay and MTT assay refer to the low DNA damage of compound 26 and 29, but good cytotoxicity of 29 to the mitochondrion cell. On the contrary, compound 26 is potent to PC-3 cell but poor to HL-60 cell. Thererfore, the two compounds will be needed to go further research. Above all, the Comet assay and MTT assay could provide an alternative method for screening the potential drugs. Part Ⅱ The morphine’s ACNO ring system derivatives have been synthesized in our lab. On chemical aspect, a series of synthetic scheme had been reported and during which, (R)-5-Hydroxy-5,6,7,8-tetrahydroisoquinoline with optical pure is an important intermediate. To get this compound, synthesis of it’s precursor, 5-Oxo-5,6,7,8-tetrahydroisoquinoline is very important. Researchers have been discovered that this compound often has low yield and usually accompanied a byproduct, 8-Oxo-5,6,7,8-tetrahydroisoquinoline. In the past, this compound was synthesized through four steps in our synthetic strategy, and the overall yield was 43%. Violent strong oxidant was used in this strategy. Therefore, to improve the strategy and raise the overall yield of this compound, we studied two references by K. C. Nicolau and P. Lardenois. The strategy of Nicolau is to use a mild oxidant, o-iodoxybenzoic acid (IBX) to undergo the direct oxidation at the carbon adjacent to aromatic ring to get the desired ketone product. Since to monitor the synthetic process with TLC is difficult, we used the High Performance Liquid Chromatography (HPLC) to find out the time that maximum product given. And then, we changed different reaction conditions and optimize to a best one. Besides, other adducts formed during the reaction process were identified by LC-MS. However, the yield of product was not correspondence with paper of that reported, and just only 26%. Hence, we tried the earlier P. Lardenois’method to solve the problem. Finally, we took the strategy of P. Lardenois and utilized HPLC to monitor the reaction as well and then confirmed the product of each step. The pure valuable ketone product was afforded after distillation. In conclusion, the yield of the first step was 78% and 91% of the second step, and the overall yield of this ketone compound was 71%. The product was scaled up to 100 g and thus could promote the final yield of the morphine ACNO derivatives. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/24778 |
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Appears in Collections: | 藥學系 |
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