以鉀離子通道促進劑cromakalim為引導化合物，在本研究中合成了一系列具有光學活性的六氫-苯駢喃[3,4-b][1,4]的衍生物（(+)-37a-c，(-)-38f-o及(-)-52），並以老鼠肝門靜脈和膀胱逼尿肌進行活性測試， 觀察其是否有抑制收縮的活性。

目標化合物中的(+)-37a-c為(4aR,10bS)-1,2,3,4a,5,10b-六氫-5,,5-二甲基-1-苯駢喃[3,4-b][1,4]-9((+)-41)的醯化產物，合成起始物環氧化合物(-)- 39是由6-氰基-2,2-二甲基-2H-1-苯駢喃經以Mn(salen)為催化劑的不齊氧化反應而來。接著以乙二醇胺( ethanolamine) 進行開環反應， 得到雙醇化合物(+)-40 。 再對雙醇中間體進行Mitsunobu 分子內環化， 則得到三環中間體 (+)-41 。化合物41分別與甲酸乙酸，丙酸和異丁醯氯反應則得到37a-c 。 本實驗室先前之研究發現(4aR,10bR)-N-苯甲醯1,2,3,4a,5,10b-六氫-5,,5-二甲-1-苯駢喃[3,4-b][1,4]-9(38e) 對膀胱具有選擇性的抑制收縮的活性。為了探討苯環上取代基的效應， 乃合成了化合物(-)-38f-o，化合物38f-n是由其(4a,10b)反式異構物((+)-50f-n)經鹼催化反應而得，化合物(-)-38o則是經由(-)-38j以三溴化硼進行去甲基化反應而得。目標化合物(-)-52為(-)-38e的類似物其合成是經由中間體(-)38b用鹽酸將其乙醯基水解後得到類化合物(-)-46，再與苯醯氯進行醯化反應而得。


依藥理活性結果38e對膀胱活性為 8.15 mM其選擇性(IC50,門靜脈/ IC50,膀胱 = 4.2倍) 。化合物38f-o在苯環上之不同取代基其對鉀離子通道活性有顯著影響，其中間位有甲基取代的38g對肝門靜脈有最高的活性(0.54mM)及選擇性( IC50 rato = 0.002) 。而對位有含未鍵接電子取代者對膀胱逼尿肌的活性較好， 如4-溴衍生物38k ( 2.0 mM )及4-基衍生物38o (3.8 mM)，38o之選擇性最高(IC50 ratio = 19.6)，苯磺化合物52在膀胱之活性僅比38e略低(11.6 mM)但選擇性較高(IC50 ratio =10.3)。

As rigid analogs of the prototype potassium channel opener (POC) cromakalim, a series of optically active hexahydro benzopyrano[3,4-b][1,4] oxazine derivatives have been synthesized. The PCO activity of target compounds were evaluated in vitro with preparations of spontaneous contracting rat portal vein and KCl-stimulated rat detrusor strip.

Target compounds (+)-37a-c are acylation products of (4aR,10aS)-1,2,3,4a,5,10b-hexahydro-5,5-dimethyl-1-benzopyrano[3,4-b][1,4] oxazine-9-carbonitrile ((+)-41). The starting epoxide (-)-39, prepared form 6-cyano-2,2-dimethyl-2H-1-benzopyran via Mn(salen) catalyzed asymmetric epoxidation, was subjected to ring-opening by 2-aminoethanol to give the diol intermediate (+)-40. Compound (+)-40 underwent intramoleculor cyclization under Mitsunobu condition to form the tricyclic intermediate (+)-41. Acylation of 41 with acetic formic anhydride, propionic anhydride, and isobutyryl chloride respectively, provided the target compounds (+)-37a-c. Earlier studies in our laboratory resulted in the identification of (4aR,10aR)-N-benzoxyl-1,2,3,4a,5,10b-hexahydro-5,5-dimethyl-1-benzopyrano[3,4-b][1,4]oxazine-9-carbonitrile ((-)-38e) as a bladder-selective potassium channel opener. To study the effects on PCO activity of different substitutents on the benzene ring of 38e, compounds (-)-38f-o were synthesized. Compound 38f-n were prepared from their corresponding (4aS,10aR)- or trans isomers (+)-50f-n. Compound 38o was obtained form 38j via O-demethylation with boron tribromide. Compound 52, the sulfonamide analog of amide(-)-38e, was obtained form 38b via hydrolysis with HCl and the subsequent reaction with benzenesulfonyl chloride.

Among the target compounds, 38g with a meta-methyl - substituent showed the highest potency (IC50 = 0.54 mM ) and selectivity (IC50, portal vein / IC 50,bladder =0.002) on the portal vein. Compounds with para-substituents having lone-pair electrons showed improved activity on the bladder, such as the 4-bromo analog 38k (IC50,bladder = 2.0 mM; IC50 ratio = 4 ) and the 4-OH analog 38o (IC50,bladder = 3.8 mM; IC50 ratio = 19.6). Benzenesulfonamide 52 was found to be bio-isosteric with benzamide 38e, with slightly lower IC50 (11.6 mM) and higher selectivity (IC50 ratio = 10.3).

中文摘要 1
英文摘要 3
一‧背景及動機 6
二‧結果與討論 26
化學合成 26
藥理 39
三‧結論 43
四‧實驗部分 45
溶劑及藥品來源 45
一般儀器及方法 46
合成步驟 49
藥理實驗 79
五‧參考文獻 81
六‧附圖 89
圖 目 錄


圖1: 鉀離子通道促進劑與心肌或平滑肌細胞之間的電生理效應 9
圖2: Cromakalim 類似化合物之結構通式14
圖3: Cromakalim的X-光繞射晶體結構25
圖4: 化合物37a的X-光繞射晶體結構28
圖5: 化合物38k的X-光繞射晶體結構34
圖6: 化合物38n的X-光繞射晶體結構35
圖7: 51e和38e分子最低能量構形36
表 目 錄

表一:化合物37a-c及其相關化合物影響老鼠肝門靜脈和膀胱逼尿
肌收縮的藥理活性41
表二:化合物38e-f和化合物52抑制老鼠肝門靜脈和膀胱逼尿肌
縮的藥理活性42
路 徑 目 錄

路徑1: 目標化合物37a-c的合成27
路徑2: 化合物38a-e的合成30
路徑3: 化合物47的合成31
路徑4: 目標化合物38e-o的合成33
路徑5: 目標化合物52的合成之一 37
路徑6: 目標化合物52的合成之二 38

1. Brocklehurst. J.C. Urology in the elderly.1984
2. Cook, N. S. The Pharmacology of Potassium Channels and Their Therapeutic Potential. Trends Pharmacol. Sci. 1988, 9, 21-28.
3. Yoshizumi. K,; Ikeda. S.; Goto. K.; Morita. T,; Nisimura. N.; Sukamoto. T. and Yoshino. K. Synthesis and Structure-Activity Relation of Novel Phenylcyanoguanidine Derivatives as Potassium Channel Openers. Chem. Pharm. Bull.1996, 44, 2042-2050.
4. Taira, N. Similarity and Dissimilarity and Mechanism of Action between Nicorandil and Classical Nitrates: An overview. J.Cardiovasc Pharmacol.1978,10, S1-9
5. Steinberg, M. I.; Robertson, D. W. The Body*s Potassium Channels. CHEMTECH 1990, 432-438.
6. Noma, A. ATP-regulated K+ Channels in Cardiac Muscle. Nature 1983, 305, 147-148.
7. Spruce, A. E. ; Standen, N. B.; Stanfield, P. R. Studies of the Unitary Properties of Adenosine-5*-Triphosphate-Regulated Potassium Channels of Frog Skeletal Muscle. J. Physiol. 1987, 382, 213-236.
8. Cook, D. L.; Hales, C. N. Intracellular ATP directly Blocks K+ Channels in Pancreatic b-cells. Nature 1984, 311, 271.
9. Wang, W.; Sackin, H.; Giebisch, G. Renal Potassium Channels and Their Regulation. Ann. Rev. Physiol. 1992, 54, 81-96.
10. Ben-Ari, Y.; Krnjevic, K.; Crepel, V. Activators of ATP-Sensitive K+ Channels Reduce Anoxic Depolarization in CA3 Hippocalpal Neurons. Neuroscience 1990, 37, 55-60.
11. Bernardi, H.; Fosset, M.; Lazdunsk, M. Characterization, Purification and Affinity Labeling of The Brain [3H] Glibenclamide-Binding Protein, A putative Neuronal ATP-Regulated K+ Channel. Proc. Natl. Acad. Sci. USA 1988, 85, 9816-9820
12. Atwal, K. S. Advances in The Structure-Activity Relationships, Mechanisms of Action, and Therapeutic Utilitres of ATP-Sensitive Potassium Channel Openners. Drug Development Res. 1994, 33, 250-262.
13. Special Issue on ATP Sensitive Potassium Channels. Cardiovasc. Res. 1984, 28, 726-930.
14. Evans, J. M.; Fake, C. S.; Hamilton, T. C.; Poyser, R. H. and Watts, E. A. Synthesis and Antihypertensive Activity of Substituted trans-4-Amino-3,4-dihydro-2,2-dimethyl-2H-1-benzopyran-3-ol. J. Med. Chem. 1983, 26, 1582-1589.
15. Evans, J. M.; Fake, C. S.; Hamilton, T. C.; Poyserr, R. H.; Showell, G. A. Synthesis and Antihypertensive Activity of 6,7-Disubstituted trans-4-Amino-3,4-dihydro-2, 2-dimethyl-2H-1-benzopyran-3-ol. J. Med. Chem. 1984, 27, 1127-1131.
16. Ashwood, V. A.; Buckingham, R. E.; Cassidy, F.; Evanss, J. M.; Faruk, E. A.; Hamilton, T. C.; Nash, D. J.; Stemp, G.; Willcocks, K. Synthesis and Antihypertensive Activity of 4-(Cyclicamido)-2H-1-benzopyrans. J. Med. Chem. 1986, 29, 2194-2201.
17. Hamiltonm T. C.; Weir, S. W. and Weston, A. H. Comparison of The Effects of BRL 34915 and Verapamil on Electrical And Mechanical Activity in Rat Portal Vein. Br. J. Pharmac. 1986, 88, 103-111.
18. Zografos, P.; Li, J. H.; Kau, S. T. Comparison of The in Vitro Effects of K+ Channel Modulators on Detrusor and Portal Vein Strips form Guinea Pigs. Pharmacol.1992, 45, 216-230
19. Ashwood, V. A.; Cassidy, F.; Coldwell, /m. C.; Evanss, J. M.; Hamilton, T. C.; Howlett, D. R.; Smith, D. M.; Stemp, G. Synthesis and Antihypertensive Activity of 4-(Substitutedcarbonylamido)-2H-1-benzopyrans. J. Med. Chem. 1990, 33, 2667-2672.
20. Burrell, G.; Cassidy, F.; Evans, J. M.; Lightowler, D.; Stemp, G. Variation in the Aromatic Ring of cromakalim : Antihypertensive Activity of Pyranopyridines and 6-Alkyl-2H-1-benzopyrans. J. Med. Chem. 1990, 33, 3023-3027.
21. Atwal, K. S.; Ferrara, F. N.; Ding, C. Z.; Grover, G. J,; Sleph, P. G.; Dzwonczyk, S,; Baird, A. J. and Normandin, D. E. Cardioselective Antiischemic ATP-Sensitive Potassium Channel Openers. 4. Structure-Activity Studied on Benzopyranyl-cyanoguanidines : Replacement of the Benzopyran Portion. J. Med. Chem. 1996, 39, 304-313.
22. Thomas, W. A.; Whitcombe, I. W. A. Conformational Behaviour of cromakalim and Related Potassium Channel Activators. J. Chem. Soc. Chem. Commum, 1990, 528-529.
23. Dalonzo. A. J; Darbenzio. R. B.; Swter. J. C.; Hess. T. A.; Grover. G. J.; Sleph. P.G. Normandin. D. E. and Lodge. N.J. A Comparison Between the Effect of BMS-180448, a Novel K+ Channel Opener, andCromakalim in Rat and Dog. Eur. J. Pharmacol, 1995, 294, 271-280.
24. Sato, H.; Koga, H.; Ishizana, T.; Makino, T.; Kuromaru, K.; Taka, N.; Takahashi, T.; Sato, T.; Nabata, H. Bioorg. Med. Chem. Lett. 1993, 3, 2624-2627.
25. Tamura, K.; Suzuki, Y.; Yoshida, S.; Nabata, H. Action of KC-399, a Newly Synthesized Potassium Channel Opener, on Mechanical Activity and 86Rb Efflux in Rat Aorta. J. Cardiovasc. Pharmacol. 1994, 23, 220-226.
26. Uchida, W.; Hirano, Y.; Shirai, Y.; Taguchi, T.; Masuda, N.; Shibasaki, K.; Hirano, S.; Matsumoto, Y.; Tsuzuki, R.; Yanagisawa, I.; Satoh, N.; Takenaka, T. Potassium Channel-Opening and Vasorelaxant Profiles of a Novel Compound YM099 in Rat Isolated portal Vein and Rabbit Isolated Aorta. Arch. Int. Pharmacodyn. 1994, 327, 330-343.
27. Uchida, W.; Hirano, Y.; Taguchi, T.; Masuda, N.; Shiral, Y.; Satoh, N.; Takenaka, T. Cardiovascular Effect of YM099, a Novel K+ Channel Openner, in Anesthtized and Conscious Dog. Eur. J. Pharmacol, 1994, 264, 285-293.
28. Uchida, W.; Masuda, N.; Taguch, T., Shibasaki, K.; Shirai, Y.; Asano, M.; Matsumoto, Y.; Tsuzuki, R.; Fujikira, T.; Takeraka, T. Pharmacologic Profiles of YM-934, a Novel Potassium Channel Opener. J. Cardiovasc, Pharmacol. 1994, 23, 180-187.
29. Yamada, M.; Terzic, A.; Findlay, I.; Jahangir, A.; Shen, W.K.; Kurachi, Y. Antihypertensive and Cardiovascular Effect of a New Potassium Channel Opener,TCV-295, in Rats and Dogs. J. Pharmacol. Exp. Ther. 1993, 267, 1544-1553.
30. Russell, K.; Brown, F. J.; Warwick, P.; Forst, I.; Grant, T.; Howe, B.;Kau, S. T.; Li, J. H.; McLaren, F. M.; Shapiro, H. S.; Trivedi, S. A Highly Potent Series of Fluoroalkyl Benzoxazine Pyridine Potassium Channel Openers. Bioorg. Med. Chem. Lett. 1993, 3, 2727-2728
31. Burrell, G.; Evanss, J. M.; Hardley, M. S.; Hicks, F.; Stemp, G. Benzopyran Potassium Channel Activators Related To Cromakalim-Heterocyclic Amide Replacements At Position 4. Bioorg. Med. Chem. Lett. 1994, 4, 1285-1290
32. Damiano, B. P.; Giardino, E. C.; Haertlein, B. J.; Stump, G. L.; Mitchell, J. A.; Falotico, R. Cardiovascular Profile of RWJ29009, a New Potassium Channel Activator, in Anesthetized and Conscious Dog. J. Cardiovasc. Pharmacol. 1994, 23, 300-310.
33. Katz, L. B.; Giardino, E. C.; Salata, J. J.; Moore, J. B.; Falotico, R. RWJ26629, a New Potassium Channel Opener and Vascular Smooth Muscle Relaxant : APotential Antihypertensive and Antiaginal Agent. J. Pharmacol., Exp. Ther. 1993, 267, 648-656.
34. Itoh, T.; Ito, S.; Shafiq, J.; Suzuki, H. Effect of a Newly Synthesized K+ Channel Opener, Y-26763, on Noradrenaline-Induced Ca2+ Mobilization in Smooth Muscle of the Rabbit Mesenteric Artery. Br. J. Pharmacol. 1994, 111, 165-172
35. Terzic, A.; Jahangir, A.; Kurachi, Y. HOE-234, a Second Generation K+ Channel Opener, Antagonizes the ATP-Dependent Gating of Cardiac ATP-Sensitive K+ Channels.J. Pharmacol. Exp. Ther. 1994, 268, 818-825.
36. Kamei, K.; Yoshida, S.; Imagawa, J. I.; Nabatu, H.; Kuriyawa, H. Regional and Species Difference in Glyburide-Sensitive K+ Channels in Airway Smooth Muscles as Estimated frim Actions of KC-128 and Levcromakalim. Br. J. Pharmacol. 1994, 113, 889-897.
37. Kusumoto, K.; Awane, Y.; Kitayoshi. T.; Fujiwara, S.; Hashigushi, S.; Teraahita.; Shiraishi, M.; Watanabe, T. J., Pharmacologic Profiles of YM934, a Novel Potassium Channel Opener.; J. Cardiovasc. Pharmacol. 1994, 23, 180-187.
38. Smith, D. G.; Buckle, D. R.; Faller, A. and Pinto, I. Pyrrole Analogues of the Pyrrolidinone Moiety of the Potassium Channel Activator Cromakalim as Relaxant of Guinea Pig Trachealis. L. Bioorg Med. Chem. Lett., 1992, 2, 1595-1598.
39. Yoo, S. E.; Kim, S. J.; Shin, H. S.; Hong, K.W. Benzopyran Sulfoxide Derivatives as New Potassium Channel Activator. Bioorg. Med. Chem. Lett., 1993, 3, 553-554.
40. Gadwood, R. C.; Kamdar, B. V.; Cipkus Dubray, L. A.; Wolfe, M. L.; Smith, M. P.; Watt, W.; Mizsak, S. A.; Groppi, V. E. Synthesis and Biological Activity of Spirocyclic Benzopyran Imidazolone Potassium Channel Openners. J. Med. Chem. 1993, 36., 1480-1487.
41. Gadwood R. C.; Thomasco, L. M.;Groppi, V. E.; Burrnett, B. A.; Humphrey, S. J.; Smith, M. P. and Watt, W., 207th ACS National Meeting, San Diegp, CA, March 13-18, Abstract 223, 1994.
42. Almansa, C.; Gomez, L. A.; Cavalacanti, F. L.; Rodriguez, R.; Carceller, E.; Bartroli, J.; Garcia-R., J. and Forn, J. 2,2-Dialkylnaphthalen-1-ones as New Potassium Channel Activators. J. Med. Chem., 1993, 36, 2121-2133.
43. Perez-Vizcaino, F.; Casis, O.; Rodriguez, R.; Gomez, L. A.; Garcia Rafanell, J. and Tamargo, J. Effect of Novel Potassium Channel Opener, UR-8225, on Contractile Responses in Rat Isolated Smooth Muscle. Br. J. Pharmacol. 1993, 110, 1165-1171.
44. Berthelon, J-J.; Brunet, M.; Noblet, M.; Durbin, P. Guerrier, D. and Luong, T. Patent Application, WO/9413658, 1994.
45. Empfield, J. R.; Mayhugh, D.; Ohnmacht, C. J.; Frank, C. ; Grant, T.; Li, J. 4-Sulfonamidoanilide Tertiary Carbinols: A Novel Series of Potassium Channel Openers. Bioorg. Med. Chem. Lett. 1997, 7, 775-778.
46. Howe,B. B; Halterman,T. J; Yochim, C.L; Do, M. L; Pettinger, S. J ; Stow, R. B; Ohnmacht, C. J; James, K. R; Trainor, D. A; Browin, F. J; Kau,S.T. ZENECA ZD6169 : A Novel KATP Channel Opener with in Vivo Selectivity for Urinary Bladder. J. Pharmacol. Exp.Ther.1995, 274, 884-890.
47. Cassidy, F.; Evanss, J. M.; Smith, D. M.; Stemp, G.; Edge, and Williams, D. J. Conformational Analysis of the Novel Antihypertensive Agent cromakalim (BRL 34915). J. Chem. Soc. Chem. Commun. 1989, 377-378.
48. Attwood, M. R.; Jones, P. S.; Kay, P. B.; Paciorek, P. M. and Redshaw, S. The Design of a Novel Class of Potassium Channel Activiting Drugs, 2-(2,2-Dimethylbenzopyran-4-yl)-Pyridine-1-Oxides. Life Sciences, 1991, 48, 803-810.
49. Part of this work has been published. See: Cheng, C.Y.; Chiu, H. I.; Chang, M. ; Lin, Y. C.; Tsai, M. C.; Yu, H. C. Synthesis of 2,3,4a,11b-Tetrahydro-oxazino{2,3,-c}benzopyran-9-carbonitriles as ATP-Sensitive Potassium Channel Openers. Bioorg. Med. Chem. Lett. 1998, 8, 463-468.
50. Chang, M. J. Ph. D. Thesis, Synthesie and Pharmacology of Novel Benzopyran Derivatives as Potassium Channel Activators. Jan. 1997
51. Lee, N. H.; Muci, A. R. ; Jacobsen, E. N. Enantiomerically Pure Exoxychromans via Asymmetric Catalysis. Tetrahedron Lett. 1991, 32, 5055-5058.
52. Houge-Frydrych, C. S. V.; Pinto, I. L. An UmusualHydroxyl Inversion By DAST . Tetrahedron Lett. 1989, 30, 3349-3350.
53. Fukuyama, T.; Jow, C.-K.; Cheung, M. 2- and 4-Nitrobenzenesulfonamides: Exceptionally Versatile Means for Preparation of Secondary Amines and Protection of Amines. Tetrahedron Lett. 1995, 36, 6373-6380.
54. 化合物37d-e及化合物38a-e係由邱欣怡同學所製備的.
55. Quast, U.; Villhauer, E. B. The Individual Enantiomers of Cis-Coromakalim Possess K+ Channel Opening Activity. Eur. J. Pharmacol. 1993, 245, 165-171.
56. Still, W. C.; Kahn, A. Rapid Chromatographic Technique for
Preparative Separations with Moderate Resolution.
J. Org. Chem.1978, 4, 73-80.