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合成脢氧苯甲醛(CCY1a)類緣化合物做為腺甘酸環化脢活化劑

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行政院國家科學委員會專題研究計畫 成果報告

合成

氧苯甲醛

(CCY1a)類緣化合物做為腺

酸環化

活化

計畫類別: 個別型計畫 計畫編號: NSC91-2320-B-039-030- 執行期間: 91 年 08 月 01 日至 92 年 07 月 31 日 執行單位: 中國醫藥大學藥物化學研究所 計畫主持人: 黃麗嬌 報告類型: 精簡報告 處理方式: 本計畫可公開查詢

中 華 民 國 92 年 11 月 11 日

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行政院國家科學委員會專題研究計畫成果報告

合成芐氧苯甲醛(CCY1a)類緣化合物做為腺苷酸環化酶活化劑

Synthesis of Benzyloxybenzaldehyde (CCY1a) Analogs as Novel

Adenylyl Cyclase Activators

計畫編號

:

NSC 91-2320-B-039-030

執行期限

:

91 年 8 月 1 日至 92 年 7 月 31 日

主持人

:

黃麗嬌 中國醫藥大學 藥物化學研究所

中文摘要 以芐氧苯甲醛(CCY1a)為先導化合物,合 成了一系列類緣物。另外將所合成的化合物 進行抗發炎、抗過敏、抑制癌細胞增殖活性 之評估,建立了初步之構效關係。大多數的 acrylaldehyde (-CH=CHCHO)衍生物具有明顯 抗發炎、抗過敏及抑制癌細胞增殖的活性; 其中(2E)-3- [2-(benzyloxy)-5-methoxy- phenyl]acrylaldehyde,(2E)-3-{2-[(4- chlorobenzyl)oxy]-5-methoxyphenyl}- acrylaldehyde,(2E)-3-{2-[(3,5-difluoro- benzyl)-oxy]-5-methoxyphenyl}acrylaldehyde 為最具潛力的物質,可作為開發新型抗發 炎、抗過敏藥物之基本架構;(2E)- 3-{2- [(4-methoxybenzyl)-oxy]phenyl}acrylaldehyde 則針對抑制 HL-60、U937 等血癌細胞株的增 生效果最佳。 關鍵字:腺苷酸環化酶活化劑、抗過敏、抗 發炎、抗增殖活性、芐氧苯甲醛類緣物。 Abstract

Using 2-benzyloxybenzaldehyde (CCY1a) as the lead compound, a series of its analogs were synthesized. The synthesized compounds were evaluated for their anti-inflammatory,

anti-allergic and anti-proliferation activities,the

preliminary SARs in this series were established. Most of the CH=CH-CHO derivatives showed significant anti-inflammatory, anti-allergic and anti-proliferation activities. The most promising

agents were compounds (2E)-3-[2-(benzyloxy)- 5-methoxyphenyl]acrylaldehyde, (2E)-3-{2-[(4- chlorobenzyl)-oxy]-5-methoxyphenyl}acryl- aldehyde and (2E)-3-{2-[(3,5-difluorobenzyl)- oxy]-5-methoxyphenyl}acrylaldehyde, which may be used as novel structural prototypes for the development of anti-inflammatory, anti- allergic agents. On the other hand, (2E)-3-{2- [(4-methoxybenzyl)oxy]phenyl}acrylaldehyde showed significant anti-proliferation activities in human leukemia cell.

Keywords: Adenylyl cyclase activators,

anti-allergy, anti-inflammatory,

anti-proliferation, benzyloxybenzaldehyde analogs.

一、計畫緣起與目的

cAMP 為廣泛分佈於細胞間的 secondary messenger,能活化 protein kinase 及 cyclic nucleotide-gated ion channel,在細胞的訊息傳 遞上扮演非常重要的角色,與很多的生理功能 都有相當密切的關係。

細胞間的 cAMP 之含量受到 adenylyl cyclase (AC) 及 phosphodiesterase (PDE)控制, 當 cAMP 含量不足時 ATP 會受到 AC 之催化 而合成 cAMP,而過量的 cAMP 也會受到 PDE 之水解而變成 AMP,到目前為止,已有九種 adenyl cyclase 之 mammalian isozymes 以及 30 種具有 cAMP phosphodiesterase 活性之

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種 small molecule~ forskolin 被證明能直接活 化 adenylyl cyclase,由於其副作用偏多,只用於 藥理研究之工具而已,並未被應用於臨床。另 外 Cholera bacillus 之 toxin 也會引起 AC 持久

性的活化,而導至嚴重的腹瀉,也未被利用

最近本研究室開發了作用機轉獨特的 novel antiplatelet agent ~ 1-benzyl-3-(5-

hydroxymethyl-2-furyl)indazole (YC-1),其抗血 小板活性是經由 NO-independent activation of soluble guanyl cyclase (sGC)同時抑制 PDE 導 致 cGMP 含量增加而產生的,由於其作用機轉 特異,啟發了 YC-1 相關研究的熱潮,數十篇的 藥理、生化研究報告陸續被發表,最近 Bayer 藥廠 Stasch 等更開發了由 YC-1 衍生的類緣化 合物 BAY-42-2272,在美國已進入 phase I 之臨 床試驗階段。 cAMP 與 cGMP 在生化及藥理學上的重 要性是可以相提並論的。仿照 YC-1 成功的實 例,數年前我們也開始探討 adenyl cyclase 之 elevating agents。經 effects on cellular cAMP levels screening 結果發現 benzyloxy-

benzaldehyde (CCY1a)具有類似 forskolin 之活 性,進而發現 CCY1a 具有顯著的 induce cancer cell apoptosis 之作用,頗具開發潛力。本計畫即 以 CCY1a 為 lead compound,有系統的合成一 系列化合物,並探討其對 cAMP 增加之活性以 及有關之生物活性,期望能建立良好的 SAR, 發現比 CCY1a 更具開發成為新藥之潛力的新 物質。 二、研究方法 如Scheme 1所示, 主要中間體2- (substituted benzyloxy)-substituted benzaldehyde是由2-hydroxy-substituted benzaldehyde在無水碳酸鉀、碘化鉀存在下, 與substituted benzyl chloride 加熱迴流而得; 此類化合物在0 ℃、20% NaOH下,與 acetaldehyde 經aldol-condensation反應即可獲 得相對應之(2E)-3-{2-[(substituted benzyl)- oxy]-substituted phenyl}acrylaldehydes。

如Scheme 2所示,將substituted benzyl chloride在無水碳酸鉀、碘化鉀存在下,分別 和2-氰基酚,鄰甲酚,柳酸,水楊酸甲酯,水 楊醯胺,鄰氨基苯甲酸乙酯及2-氨基-4, 5-二 甲氧基苯甲酸甲酯進行benzylation即可獲得

2-substituted benzyloxy benzonitriles, 2-substituted benzyloxy 2-methylbenzenes, 2-substituted benzyloxy phenyl methanols, 2-substituted benzyloxy benzoates, 2-substituted benzyloxy benzamides和2-substituted

benzylamino benzoates。若將R1為COOR或CN

者於NaOH (aq)下進行水解反應,隨後以冰醋酸

酸化即可得到相對應之2-substituted benzyloxy benzoic acids或2-substituted benzylamino benzoic acids。

將所合成的標的化合物及其中間體進行 腺苷酸環化酶活性試驗外,並提供進行抗過 敏、抑制癌細胞增殖等活性之評估。 三、結果與討論 合成一系列2-benzyloxybenzaldehyde (CCY1a)之類緣物,包括(2E)-3-{2-

[(substituted benzyl)oxy]-substituted phenyl}- acrylaldehydes、2-substituted benzyloxy benzoic acids衍生物。部分標的化合物經anti-

inflammatory, anti-allergic 及anti-proliferation activity篩選初步建立其結構與活性之關係。 具有acrylaldehyde (-CH=CHCHO)之衍生物對 於抗發炎、抗過敏及抑制癌細胞增殖的活性 均較為顯著;其中(2E)-3-[2-(benzyloxy)- 5-methoxyphenyl]acrylaldehyde,(2E)-3-{2- [(4-chlorobenzyl)- oxy]-5-methoxyphenyl}- acrylaldehyde,(2E)- 3-{2-[(3,5-difluoro- benzyl)-oxy]-5-methoxyphenyl}acrylaldehyde 為最具潛力的物質,可作為開發新型抗發 炎、抗過敏藥物之基本架構;(2E)-3-{2-[(4- methoxybenzyl)-oxy] phenyl}-acrylaldehyde針 對抑制HL-60、U937等血癌細胞株的增生效果 最佳。2-Substituted benzyloxy benzoic acid及 其相關化合物較不具抗發炎、抗過敏活性。 此外亦發現2-[(4-methoxybenzyl)oxy] benzoic acid和2-[(4-chlorobenzyl)amino] benzoic acid 對抗下痢具有顯著的活性表現,值得進一步 探討。至於腺苷酸環化酶活性試驗仍在測試 中。

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OH CHO CH2Cl(Br) O CHO O CHO O CHO O CH=NOH CH2Cl(Br) OH CHO + K2CO3, KI Ethanol R3' R2' R4' R5' R6' R3 R5 R4 R3 R5 R4 R3' R2' R4' R5' R6' R3 R5 R4 R3' R2' R4' R5' R6' CH3CHO, 20% NaOH Ethanol R3 R5 R4 R3' R2' R4' R5' R6' CH3CHO, 20% NaOH Ethanol R3 = H, Cl R2' = H, F, Cl R5' = H, F R4 = H, OCH3 R3' = H, F, Cl R6' = H, F, Cl R5 = H, Cl, OCH3 R4' = H, F, Cl, OCH3 , CH3 Scheme 1 R5 R3' R2' R4' R5' NH2OH.HCl CH3COONa/EtOH R3' R2' R4' R5' R6' + R3 R5 R4 K2CO3, KI Ethanol

CH2 Cl X CH2 + K2CO3 , KI ethanol or methanol R1 = CN, CH3, CH2OH, COOCH3, R2' = H, Cl, F COOC2H5, CONH2 R2 = OH, NH2 R3' = H, Cl, F, OCH3 R4 = H, OCH3 R4' = H, Cl, F, OCH3 R5 = H , OCH3 X = O, NH R1 R2' R4' R5 R3' R1 R2' R3' R4' R5 Scheme 2 R4 R4 R2 三、計畫成果自評 本計畫研究結果雖有部份藥理結果仍在 測試中,整體而言,具有學術或應用價值,擬 即將發表於學術期刊。 四、參考文獻

1. J. Krupinski, F. Coussen, H. A. Bakalar, W. J. Tang, P. G. Feinstein, K. Orth, R. R. Reed and A. G. Gilman, Adenylyl cyclase amino acid sequence: possible channel- or transporter- like structure, Science, 244, 1558-1564. (1989).

2. R. K. Sunahara, C. W. Dessauer and A. G. Gilman, Complexity and diversity of

mammalian adenylyl cyclases, Annu. Rev.

Pharmacol. Toxicol., 36, 461-480. (1996).

3. M. D. Houslay and G. Milligan, Tailoring cAMP-signalling response through isoform multiplity, Trands Biochem. Sci., 22(6), 217-224. (1997).

4. F. Y. Lee, J. C.Lien, L. J. Huang, T. M. Huang, S. C. Tsai, C. M. Teng, C. C. Wu, F. C. Cheng and S. C. Kuo, Synthesis of 1-benzyl-3- (5-hydroxymethyl-2-furyl)indazole analogues as novel anti-platelet agents, J. Med. Chem.,

44(22), 3746-3749. (2001).

5. C. C. Wu, F. N. Ko, S. C. Kuo, F. Y. Lee, C. M. Teng, YC-1 inhibited human platelet aggregation through NO-indepent activation of soluble guanylate cyclase, Br. J.

Pharmacolog. 116, 1973-1978. (1995).

6. F. N. Ko, C. C. Wu, S. C. Kuo, F. Y. Lee, C. M. Teng, YC-1, a novel activator of platelet guanylate cyclase, Blood, 84, 4226-4233. (1994).

7. J. P. Stasch, E. M. Becker, C. Alonso-Aalija, H. Apeper, K. Dembowsky, et al., NO- independent regulatory site on soluble guanylate cyclase, Nature, 410, 212. (2001). 8. M. Markert, P. C. Andrews, B. M. Babior,

Measurement of O2- production by human

neutrophils. The preparation and assay of NADPH oxidase-containing particles from human neutrophils, Methods Enzymol., 105, 358-365. (1984).

9. A. C. Newby, Role of adenosine deaminase, ecto-(5'-nucleo-tidase) and ecto-(non-specific phosphatase) in cyanide-induced adenosine monophosphate catabolism in ratpolymorpho- nuclear leucocytes, Biochem. J., 186, 907-918. (1980).

10. M. W. Verghese, K. Fox, L. C. McPhail and R. Snydermen, Chemoattractant-elicited alteration of cAMP levels in human polymorphonuclear leukocytes require a

Ca2+-dependent mechanism which is

independent of transmembrane activation of adenylyl cyclase, J. Biol. Chem., 260, 6769-6775. (1985).

11. M. Saad, C. F. Strnad and K. Wong, Dual regulation of neutrophil adenylate cyclase by fluoride and its relationship to cellular activation, Br. J. Pharmacol., 91, 715-719. (1987).

12. C. D. Wright, P. J. Kuipers, D. Kobylarz-Singer, L. J. Devall, B. A.

(5)

Klinkefus and R. E. Weisharr, Differential inhibition of human neutrophil functions: role of cyclic AMP-specific, cyclic GMP-

insensitive phosphodiesterase, Biochem.

Pharmacol., 40, 699-707. (1990).

13. P. G. Grady and L. L. Thomas, Characterization of cyclic nucleotide phosphodiesterase activities in resting and

N-formylmethionyl-leucylphenyl-alanine-sti

mulated human neutrophils, Biophim.

Biophys. Acta., 885, 282-293. (1986).

14. G. Lopez-Lluch; M. I. Huron; F. J. Alcain; J. M. Quesada; P. Navas, Redox regulation of camp levels by ascorbate in 1,25-dihydroxy- vitamine d3-induced differentication of HL-60 cells. Biochem. J., 331(Pt. 1), 21-27. (1998).

15. Q. He; D. Jiang, A novel aminosteroid is

active for proliferation inhibition and differentiation induction of human acute

myeloid leukemia HL-60 cells. Leuk. Res.

23(4), 369-372. (1999).

16. M. V. Berridge; A. S. Tan, Characterization of the cellular reduction of 3-(4,5-Dimethyl- thiazol-2-yl)-2,5-diphenyltetrazolium Bromide (MTT): subcellular localization, substrate dependence, and involvement of mitochondrial electron transport in MTT reduction. Arch. Biochem. Biophys. 303, 474-482. (1993).

17. M. Markert, P. C. Andrews, B. M. Babior, Measurement of 02- production by human neutrophils. The preparation and assay of NADPH oxidase-containing particles from human neutrophils, Methods Enzymol., 105, 358-365. (1984).

18. A. C. Newby, Role of adenosine deaminase, ecto-(5'-nucleotidase) and ecto-(non-specific phosphatase) in cyanide-induced adenosine monophosphate catabolism in rat

polymorphonuclear leucocytes, Biochem. J.,

186, 907-918. (1980).

19. A. J. Barrett, "A Laboratory Handbook", Dingle, J. T. ed. (Elsevier/North-Holland, Amsterdam), 118-120. (1972).

20. D. R. Absolom, Basic methods for the study of phagocytosis, Methods Enzymol., 132, 95-179. (1986).

21. J. P Wang, S. L. Raung, M. F. Hsu and C. C. Lin, Inhibition by gomisin C (a lignan from

Schizandra chinensis) of the respiratory burst

of rat neutrophils, Br. J. Pharmacol., 113, 945-953. (1994).

22. B. Goldberg and A. Stem, The role of the superoxide anion as a toxic species in the erythrocyte, Arch. Biochem. Biophys., 178, 218-225. (1977).

23. D. E. McClain, M. A. Donlon, S. Chock and G. N. Catravas, The effect of calmodulin on histamine release in the rat peritoneal mast cell, Biochem. Biophys. Acta., 763, 419. (1983).

24. T. H. P. Hanahae, Mechanism of histamine release from rat isolated peritoneal mast cells by dextran: the role ofimmunglobulin E.,

Agents Action, 14, 468. (1984).

25. A. R. Johnson and E. G. Erdos, Release of histamine from mast cell by vasoactive peptides, Proc. Soc. Exp. Biol. Med., 142, 1252-1256. (1973).

26. R. Hakanson and A. L. Ronnberg, Improved fluorometric assay of histamine, Antlyt.

Biochem., 60, 560-567. (1974).

27. S. B. Corradin, J. Manuel, S. D. Donini, E. Quattrocchi and P. Ricciardi-Castagnoi, Inducible nitric oxide synthase activity of cloned murine microglial cells, Glia 7, 255-262. (1993).

28. L. Minghetti, A. Nicolini, E. Polazzi, C. Creminon, J. Maclouf and G. Levi, Inducible nitric oxide synthase expression in activated rat microglial culcures is down-regulated by exogenous prostaglandin E2 and by

cyclooxygenase inhibitors, Glia 19, 152-160. (1997).

參考文獻

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