臺灣博碩士論文加值系統

本論文是利用微波加速化學反應及化學選擇性控制方法，合成出1H-pyrazol-5-yl-N,N-dimethyl-formamidine和pyrazolyl-2-azadiene的兩類化合物，將5-amino-1,3-diphenyl pyrazole，1H-pyrazol-5-yl-N,N-dimethyl-formamidines，pyrazolyl-2-azadiene，5-amino-4-formylpyrazoles四類具構效關係的化合物經由藥理活性篩選 (NCI-H661，NPC-TW01，Jurkat)。

其結果顯示1H-pyrazol-5-yl-N,N-dimethylformamidines衍生物2b，2c，2d具較佳的藥理活性 (IC50: 6.0~9.2μM)，從藥理活性也指出在pyrazole derivatives須同時擁有amidinyl與formyl官能基才能增強藥理活性。

Chemoselective microwave-assisted amidination was successfully developed to alternatively synthesize 1H-pyrazol-5-yl-N,N-dimethyl-formamidine and pyrazolyl-2-azadiene two classes compounds. All of the starting materials and resulting products were tested against NCI-H226, NPC-TW01, and Jurkat cancer cell lines to evaluate their difference in antiproliferative activities for realizing the structure activity relationship study.

Following the SAR result, 1H-pyrazol-5-yl-N,N-dimethylformamidine compounds 2b, 2c and 2d possessed the best potent with IC50 values in low micromolar range. On the other hand, we found that the formyl group at C-4 position and the grafted amidinyl group in the main core of pyrazolic molecule were necessary for the inhibitory activity

Chart List................................................III
Abbreviation List........................................VI
中文摘要.................................................VII
Abstract..................................................VIII
Chapter 1 Introduction.....................................1
Section 1.1 Pyrazoles derivatives as Antibacterial Lead Compound...................................................2
Section 1.2 Pyrazoles derivatives as Antifungal Lead Compounds..................................................6
Section 1.3 Pyrazoles derivatives as Anticancer Lead Compounds..................................................9
Section 1.4 Improved bioactivities by Amidinyl Group.....................................................12
Section 1.5 Antiproliferative of Formylated Amidinyl Pyrazole derivatives......................................14
Chapter 2　Research Approach..............................15
Chapter 3　Result and Discussion..........................17
Section 3.1 Chemistry....................................17
Section 3.1.1 Synthesis of 5-amino-1,3-diphenyl pyrazole (1a–1e)...................................................18
Section 3.1.2 Synthesis of 1H-pyrazol-5-yl-N,N-dimethylformamidines (2a–2e)..............................20
Section 3.1.3 Synthesis of Pyrazolyl-2-azadienes
(3a–3e)...................................................22
Section 3.1.4 Synthesis of the de-amidination of methnimidamide (4a–4e)....................................27
Section 3.2 Biological evaluations........................29
Chapter 4　Conclusion.....................................35
Chapter 5　Experimental Section...........................36
Section 5.1 General Procedure.............................36
Section 5.2 Spectrum......................................39
Section 5.3 Cell lines....................................50
Section 5.4 Growth inhibition assay.......................51
Reference.................................................52
Addendum..................................................59

1. (a) Kim, G. H., [Extraesophageal manifestations of gastroesophageal reflux disease]. Korean J. Gastroenterol. 2008, 52, 69; (b) Morales, M. C.; Basomba, A.; Villalmanzo, I. G.; Pelaez, A.; Campos, A.; Guerrero, M., Crossed tolerance to pyrazolines, mephenamic acid, and glafenine in A. S. A.-triad patients desensitized to aspirin. J. Allergy Clin. Immunol. 1985, 75, 528.
2. (a) Tanitame, A.; Oyamada, Y.; Ofuji, K.; Fujimoto, M.; Suzuki, K.; Ueda, T.; Terauchi, H.; Kawasaki, M.; Nagai, K.; Wachi, M.; Yamagishi, J., Synthesis and antibacterial activity of novel and potent DNA gyrase inhibitors with azole ring. Bioorg. Med. Chem. 2004, 12, 5515; (b) Finn, J.; Mattia, K.; Morytko, M.; Ram, S.; Yang, Y.; Wu, X.; Mak, E.; Gallant, P.; Keith, D., Discovery of a potent and selective series of pyrazole bacterial methionyl-tRNA synthetase inhibitors. Bioorg. Med. Chem. Lett. 2003, 13, 2231.
3. (a) Rovnyak, G. C.; Millonig, R. C.; Schwartz, J.; Shu, V., Synthesis and antiinflammatory activity of hexahydrothiopyrano[4,3-c]pyrazoles and related analogues. J. Med. Chem. 1982, 25, 1482; (b) Nagakura, M.; Ota, T.; Shimidzu, N.; Kawamura, K.; Eto, Y.; Wada, Y., Syntheses and antiinflammatory actions of 4,5,6,7-tetrahydroindazole-5-carboxylic acids. J. Med. Chem. 1979, 22, 48; (c) Sugiura, S.; Ohno, S.; Ohtani, O.; Izumi, K.; Kitamikado, T.; Asai, H.; Kato, K., Synthesis and antiinflammatory and hypnotic activity of 5-alkoxy-3-(N-substituted carbamoyl)-1-phenylpyrazoles. J. Med. Chem. 1977, 20, 80.
4. (a) Ouyang, G.; Cai, X. J.; Chen, Z.; Song, B. A.; Bhadury, P. S.; Yang, S.; Jin, L. H.; Xue, W.; Hu, D. Y.; Zeng, S., Synthesis and antiviral activities of pyrazole derivatives containing an oxime moiety. Journal of Agricultural and Food Chemis ry 2008, 56, 10160; (b) Bandgar, B. P.; Gawande, S. S.; Bodade, R. G.; Gawande, N. M.; Khobragade, C. N., Synthesis and biological evaluation of a novel series of pyrazole chalcones as anti-inflammatory, antioxidant and antimicrobial agents. Bioorg. Med. Chem. 2009, 17, 8168.
5. (a) Balbi, A.; Anzaldi, M.; Maccio, C.; Aiello, C.; Mazzei, M.; Gangemi, R.; Castagnola, P.; Miele, M.; Rosano, C.; Viale, M., Synthesis and biological evaluation of novel pyrazole derivatives with anticancer activity. European journal of medicinal chemistry 2011, 46, 5293; (b) Neelarapu, R.; Holzle, D. L.; Velaparthi, S.; Bai, H.; Brunsteiner, M.; Blond, S. Y.; Petukhov, P. A., Design, synthesis, docking, and biological evaluation of novel diazide-containing isoxazole- and pyrazole-based histone deacetylase probes. J. Med. Chem. 2011, 54, 4350.
6. (a) Diana, G. D.; Carabateas, P. M.; Williams, G. L.; Pancic, F.; Steinberg, B. A., Synthesis and antiherpetic activity of some 4-[(aryloxy)alkyl]pyrazoles. J. Med. Chem. 1981, 24, 731; (b) Gudmundsson, K. S.; Johns, B. A.; Wang, Z.; Turner, E. M.; Allen, S. H.; Freeman, G. A.; Boyd, F. L., Jr.; Sexton, C. J.; Selleseth, D. W.; Moniri, K. R.; Creech, K. L., Synthesis of novel substituted 2-phenylpyrazolopyridines with potent activity against herpesviruses. Bioorg. Med. Chem. 2005, 13, 5346; (c) Moukha-chafiq, O.; Taha, M. L.; Lazrek, H. B.; Vasseur, J. J.; De Clercq, E., Synthesis and biological evaluation of some acyclic alpha-(1H-pyrazolo-[3,4-d]pyrimidin-4-yl)thioalkylamide nucleosides. Nucleosides Nucleotides Nucleic Acids 2002, 21, 165.
7. (a) Ozdemir, Z.; Kandilci, H. B.; Gumusel, B.; Calis, U.; Bilgin, A. A., Synthesis and studies on antidepressant and anticonvulsant activities of some 3-(2-furyl)-pyrazoline derivatives. European journal of medicinal chemistry 2007, 42, 373; (b) Kornet, M. J.; Garrett, R. J., Synthesis of 1-phenyl-2-(phenylcarbamoyl)pyrazolidines as potential anticonvulsant agents. J. Pharm. Sci. 1979, 68, 377; (c) Kornet, M. J., Synthesis of 1-methyl-2-phenylcarbamoylpyrazolidines as potential anticonvulsant agents. J. Pharm. Sci. 1978, 67, 1471.
8. (a) Farghaly, A. M.; Habib, N. S.; Khalil, M. A.; el-Sayed, O. A., Synthesis of some thiazole-, 1,3,4-thiadiazole-, and 4H-1,2,4-triazole derivatives of pyrazolo[3,4-b]quinoline. Arch. Pharm. 1991, 324, 19; (b) Senga, K.; Novinson, T.; Springer, R. H.; Rao, R. P.; O''Brien, D. E.; Robins, R. K.; Wilson, H. R., Synthesis and antitrichomonal activity of certain pyrazolo (1,5-a) pyrimidines. J. Med. Chem. 1975, 18, 312; (c) Bekhit, A. A.; Fahmy, H. T.; Rostom, S. A.; Baraka, A. M., Design and synthesis of some substituted 1H-pyrazolyl-thiazolo[4,5-d]pyrimidines as anti-inflammatory-antimicrobial Agents. European journal of medicinal chemistry 2003, 38, 27.
9. (a) Huang, Y. R.; Katzenellenbogen, J. A., Regioselective synthesis of 1,3,5-triaryl-4-alkylpyrazoles: novel ligands for the estrogen receptor. Org Lett 2000, 2, 2833; (b) Kraus, G. A.; Bae, J., Synthesis of N-(2-methylpropyl)-2E-undecene-8,10-diynamide, a novel constituent of Echinacea angustifolia. Tetrahedron Lett. 2003, 44, 5505.
10. Sakya, S. M.; Rast, B., Efficient synthesis of 5-alkyl amino and thioether substituted pyrazoles. Tetrahedron Lett. 2003, 44, 7629.

11. Genin, M. J.; Biles, C.; Keiser, B. J.; Poppe, S. M.; Swaney, S. M.; Tarpley, W. G.; Yagi, Y.; Romero, D. L., Novel 1,5-diphenylpyrazole nonnucleoside HIV-1 reverse transcriptase inhibitors with enhanced activity versus the delavirdine-resistant P236L mutant: lead identification and SAR of 3- and 4-substituted derivatives. J. Med. Chem. 2000, 43, 1034.
12. Dueweke, T. J.; Pushkarskaya, T.; Poppe, S. M.; Swaney, S. M.; Zhao, J. Q.; Chen, I. S.; Stevenson, M.; Tarpley, W. G., A mutation in reverse transcriptase of bis(heteroaryl)piperazine-resistant human immunodeficiency virus type 1 that confers increased sensitivity to other nonnucleoside inhibitors. Proc. Natl. Acad. Sci. U. S. A. 1993, 90, 4713.
13. (a) Wachi, M.; Iwai, N.; Kunihisa, A.; Nagai, K., Irregular nuclear localization and anucleate cell production in Escherichia coli induced by a Ca2+ chelator, EGTA. Biochimie 1999, 81, 909; (b) Hiraga, S.; Niki, H.; Ogura, T.; Ichinose, C.; Mori, H.; Ezaki, B.; Jaffe, A., Chromosome partitioning in Escherichia coli: novel mutants producing anucleate cells. J. Bacteriol. 1989, 171, 1496.
14. Tanitame, A.; Oyamada, Y.; Ofuji, K.; Fujimoto, M.; Iwai, N.; Hiyama, Y.; Suzuki, K.; Ito, H.; Terauchi, H.; Kawasaki, M.; Nagai, K.; Wachi, M.; Yamagishi, J., Synthesis and antibacterial activity of a novel series of potent DNA gyrase inhibitors. Pyrazole derivatives. J. Med. Chem. 2004, 47, 3693.
15. (a) Gold, H. S.; Moellering, R. C., Jr., Antimicrobial-drug resistance. New Engl. J. Med. 1996, 335, 1445; (b) Payne, D. J.; Wallis, N. G.; Gentry, D. R.; Rosenberg, M., The impact of genomics on novel antibacterial targets. Current opinion in drug discovery & development 2000, 3, 177.
16. (a) Schimmel, P.; Tao, J.; Hill, J., Aminoacyl tRNA synthetases as targets for new anti-infectives. FASEB J. 1998, 12, 1599; (b) Tao, J.; Schimmel, P., Inhibitors of aminoacyl-tRNA synthetases as novel anti-infectives. Expert opinion on investigational drugs 2000, 9, 1767.
17. Hughes, J.; Mellows, G., On the mode of action of pseudomonic acid: inhibition of protein synthesis in Staphylococcus aureus. J. Antibiot. 1978, 31, 330.
18. (a) Gallant, J. E., Antiretroviral therapy in naive patients. The Hopkins HIV report : a bimonthly newsletter for healthcare providers / Johns Hopkins University AIDS Service 2000, 12 , 1, 6; (b) Yu, X. Y.; Hill, J. M.; Yu, G.; Yang, Y.; Kluge, A. F.; Keith, D.; Finn, J.; Gallant, P.; Silverman, J.; Lim, A., A series of quinoline analogues as potent inhibitors of C. albicans prolyl tRNA synthetase. Bioorg. Med. Chem. Lett. 2001, 11, 541.
19. (a) Blanquet, S.; Fayat, G.; Poiret, M.; Waller, J. P., The mechanism of action of methionyl-tRNA synthetase from Escherichia coli. Inhibition by adenosine and 8-aminoadenosine of the amino-acid activation reaction. Eur. J. Biochem. 1975, 51, 567; (b) Lee, J.; Kang, M. K.; Chun, M. W.; Jo, Y. J.; Kwak, J. H.; Kim, S., Methionine analogues as inhibitors of methionyl-tRNA synthetase. Bioorg. Med. Chem. Lett. 1998, 8, 3511; (c) Jarvest, R. L.; Berge, J. M.; Berry, V.; Boyd, H. F.; Brown, M. J.; Elder, J. S.; Forrest, A. K.; Fosberry, A. P.; Gentry, D. R.; Hibbs, M. J.; Jaworski, D. D.; O''Hanlon, P. J.; Pope, A. J.; Rittenhouse, S.; Sheppard, R. J.; Slater-Radosti, C.; Worby, A., Nanomolar inhibitors of Staphylococcus aureus methionyl tRNA synthetase with potent antibacterial activity against gram-positive pathogens. J. Med. Chem. 2002, 45, 1959.
20. Li, Y.; Zhang, H. Q.; Liu, J.; Yang, X. P.; Liu, Z. J., Stereoselective synthesis and antifungal activities of (E)-alpha-(methoxyimino)benzeneacetate derivatives containing 1,3,5-substituted pyrazole ring. Journal of Agricultural and Food Chemistry 2006, 54, 3636.
21. (a) Prakash, O.; Kumar, R.; Sehrawat, R., Synthesis and antibacterial activity of some new 2,3-dimethoxy-3-hydroxy-2-(1-phenyl-3-aryl-4-pyrazolyl)chromanones. European journal of medicinal chemistry 2009, 44, 1763; (b) Prakash, O.; Kumar, R.; Parkash, V., Synthesis and antifungal activity of some new 3-hydroxy-2-(1-phenyl-3-aryl-4-pyrazolyl) chromones. European journal of medicinal chemistry 2008, 43, 435.
22. Bawaskar, H. S.; Joshi, S. R., Organophosphorus poisoning in agricultural India--status in 2005. J. Assoc. Physicians India 2005, 53, 422.
23. de Ruijter, A. J.; van Gennip, A. H.; Caron, H. N.; Kemp, S.; van Kuilenburg, A. B., Histone deacetylases (HDACs): characterization of the classical HDAC family. Biochem J 2003, 370, 737.
24. Acharya, M. R.; Sparreboom, A.; Venitz, J.; Figg, W. D., Rational development of histone deacetylase inhibitors as anticancer agents: a review. Mol Pharmacol 2005, 68, 917.
25. Korner, M.; Tibes, U., Histone deacetylase inhibitors: a novel class of anti-cancer agents on its way to the market. Prog. Med. Chem. 2008, 46, 205.
26. Oehme, I.; Deubzer, H. E.; Wegener, D.; Pickert, D.; Linke, J. P.; Hero, B.; Kopp-Schneider, A.; Westermann, F.; Ulrich, S. M.; von Deimling, A.; Fischer, M.; Witt, O., Histone deacetylase 8 in neuroblastoma tumorigenesis. Clin. Cancer. Res. 2009, 15, 91.
27. Witt, O.; Deubzer, H. E.; Milde, T.; Oehme, I., HDAC family: What are the cancer relevant targets? Cancer Lett. 2009, 277, 8.
28. Bielawski, K.; Bielawska, A.; Sosnowska, K.; Miltyk, W.; Winnicka, K.; Palka, J., Novel amidine analogue of melphalan as a specific multifunctional inhibitor of growth and metabolism of human breast cancer cells. Biochem. Pharmacol. 2006, 72, 320.
29. Busko-Oszczapowicz, I.; Cieslak, J., [Semisynthetic penicillins. X. Synthesis of 6-formami-dinopenicillanic acid derivatives of cyclic secondary amines]. Acta Pol. Pharm. 1973, 30, 43.
30. Marcinkowska, E.; Ziolkowski, P.; Pacholska, E.; Latos-Grazynski, L.; Chmielewski, P.; Radzikowski, C., The new sensitizing agents for photodynamic therapy: 21-selenaporphyrin and 21-thiaporphyrin. Anticancer Res. 1997, 17, 3313.
31. Felmingham, D.; Robbins, M. J.; Ingley, K.; Mathias, I.; Bhogal, H.; Leakey, A.; Ridgway, G. L.; Gruneberg, R. N., In-vitro activity of trovafloxacin, a new fluoroquinolone, against recent clinical isolates. J. Antimicrob. Chemother. 1997, 39, 43.
32. Cheng, K. M.; Huang, Y. Y.; Huang, J. J.; Kaneko, K.; Kimura, M.; Takayama, H.; Juang, S. H.; Wong, F. F., Synthesis and antiproliferative evaluation of N,N-disubstituted-N''-[1-aryl-1H-pyrazol-5-yl]-methnimidamides. Bioorg. Med. Chem. Lett. 2010, 20, 6781.
33. Pinkerton, A. B.; Huang, D.; Cube, R. V.; Hutchinson, J. H.; Struthers, M.; Ayala, J. M.; Vicario, P. P.; Patel, S. R.; Wisniewski, T.; DeMartino, J. A.; Vernier, J. M., Diaryl substituted pyrazoles as potent CCR2 receptor antagonists. Bioorg. Med. Chem. Lett. 2007, 17, 807.
34. Antilla, J. C.; Baskin, J. M.; Barder, T. E.; Buchwald, S. L., Copper-diamine-catalyzed N-arylation of pyrroles, pyrazoles, indazoles, imidazoles, and triazoles. J. Org. Chem. 2004, 69, 5578.
35. Kempfer, H., How to choose a correspondence school; a guide for youth, adults, and counselors. Bellman Pub. Co.: Cambridge, Mass., 1959; p 35.