| The significance of the dihydropyrazole heterocycle is evidenced by its presence as a structural subunit in a variety of pharmacologically interesting compounds. The naturally occurring biologically active flavonoids and isoflavonoids and suitably substituted a, P-unsaturated ketones may serve as ideal constituents to access dihydropyrazole, a small bioactive molecule prevalent in numerous pharmaceutically active compounds. Since dihydropyrazole compounds were most widely used in organic synthesis and other areas, so build a structure of dihydropyrazole heterocyclic system is of great significance. Based drug design principles, found in the2H-pyrazole appropriate place to introduce polycyclic coumarin skeleton is conducive to the formation of a stable conformation, the activity will be improved. Similarly, coumarin compound is an important class of organic heterocyclic compounds with antibacterial, disinfectant, anticoagulant and anti-tumor and other physiological activity, which has become a hot drug research. Structure-based design principles, by means of molecular simulation techniques, a total of115novel compounds, that is nine novel series of aryl dihydropyrazole moiety and dihydropyrazole containing coumarin moiety were designed、synthesized and screened structure-activity relationship in this paper. All of the compounds were characterized by1H-NMR,13C-NMR, elemental analysis and other modern means of physical analysis, among them,30compounds were provided with single crystal X-ray structural analysis. Many of which compounds have good anti-bacterial, anti-cancer activity. The results were concluded as follows.(1) A total of20acetyl1-acetyl-5-aryl-2H-pyrazole derivatives were synthesized and characterized by1H NMR,13C-NMR and elemental analysis in this part (1,2,3a～3b,4g～4t,5b,6). Compounds2,4h,4q,4s,5b and6were provided with single crystal X-ray structural analysis. In order to obtain new and efficient antimicrobial lead compounds, the aromatic hydroxyl was modified, changing the side chain structure. All of the compounds have been screened for their antibacterial potential in vitro against B. subtilis ATCC6633, E. coli ATCC35218, P. fluorescens ATCC13525, and S. aureus ATCC6538. The MICs of the test compounds were determined by a colorimetric method using the dye MTT. The results showed that compound5b showed antibacterial activity against P. fluorescens with MIC of0.39μg/mL, which was even better than that of the commercial novobiocin (0.78μg/mL), compound6showed antibacterial activity against B. subtilis with the MIC of1.562ug/mL, which was even better than that of the commercial penicillin (6.25μg/mL). Structure-activity relationship studies showed that acetyl chloride transformation is conducive to the improvement of antimicrobial activity. (2) A total of28benzoyl (sulfone)-aryl-2H-pyrazole derivatives (7a～7v,8a～8g) as potential telomerase inhibitors were synthesized and characterized by1H NMR,13C-NMR and elemental analysis in this part. Compounds7a,7d,7e,7f,7h,7j,7l,7m,7n,7o,7q,7p,7t,7u,8c and8d were provided with single crystal X-ray structural analysis. All compounds were screened for anticancer activity of PC3, Bcap-37and MGC-803. The IC50of the test compounds were determined by a colorimetric method using the dye MTT. The compound7h showed anticancer activities against PC3, Bcap-37and MGC-803with the IC50of4.01±0.21,6.37±0.55,4.87±0.61μM respectively, comparable to that of positive control AMD (0.87±0.10,1.12±0.09,1.01±0.08μM respectively), compound7q showed anticancer activities against PC3and MGC-803with the IC50of5.82±1.00,4.91±0.77μM respectively, comparable to that of positive control AMD (0.87±0.10,1.01±0.08μM respectively). The selected compounds were assayed for telomerase inhibition by a modified TRAP assay, the results show that the compound7q with high anticancer activity of cell showed activity against telomerase with the IC50of4.17±0.25μM, comparable to that of positive control Ethidium bromide (2.6±0.7μM). The study on structure-activity relationships of these derivatives indicated that transformation of acetyl can improve the antitumor activity.(3) A total of312-(amino)4,5-2H-pyrazole derivatives (9a～9q,10a～10n) as potential telomerase inhibitors were synthesized and characterized by1H NMR,13C-NMR and elemental analysis in this part. Compounds9d,9g,9k,91,9n were provided with single crystal X-ray structural analysis. All compounds were screened for anticancer activity of PC3, Bcap-37and MGC-803. The IC50of the test compounds were determined by a colorimetric method using the dye MTT. Some of the compounds showed good anti-cancer activity. Among them, compounds9n,10e,10f,10h, lOn showed anticancer activities against PC3with the IC50of3.44±0.46,4.07±0.35,3.11±0.11,3.67±0.22,3.58±0.11μM, comparable to that of positive control AMD (0.87±0.10μM), compounds9b,9d,10h showed anticancer activities against Bcap-37with the IC50of4.96±0.99,5.35±0.55,4.07±0.18μM, comparable to that of positive control AMD (1.12±0.09μM), compounds9c,9i,10i,10j, lOn showed anticancer activities against MGC-803with the IC50of4.31±0.78,4.81±0.68,3.51±0.21,4.12±0.11,3.01±0.13μM respectively, comparable to that of positive control AMD (1.01±0.08μM). According to preliminary anticancer activity of structure-activity relationship, acetylation of pyrazole skeleton5-aryl substituent will improve the anticancer activity, the more straight-chain amino ring amine is more conducive to activity. The selected compounds were assayed for telomerase inhibition by a modified TRAP assay, the results show that the compound10h with high anticancer activity of PC3, Bcap-37cells showed activity against telomerase with the IC50of3.98±0.22μM, comparable to that of positive control Ethidium bromide (2.63±0.7μM). Certain degree of consistency of cytotoxic activity of compounds with activity of telomerase, to reveal these compounds are expected to be effective telomerase inhibitors.(4) A total of36acid (sulfone) base2H-pyrazole-coumarin skeleton compounds (11a～11j,12k～121,13a～13x) as potential telomerase inhibitors were synthesized and characterized by1H NMR,13C-NMR and elemental analysis in this part. Compounds13f,13k,13x were provided with single crystal X-ray structural analysis. All compounds were screened for anticancer activity of PC3, Bcap-37and MGC-803. The IC50of the test compounds were determined by a colorimetric method using the dye MTT. Some of the compounds showed good anti-cancer activity. Among them, compounds11i,13c,13f showed anticancer activities against PC3with the IC50of2.98±0.15,1.87±0.11,1.92±0.13μM, comparable to that of positive control AMD (0.87±0.10μM), compounds11j,13c,13i showed anticancer activities against Bcap-37with the IC50of6.30±0.25,2.21±0.10,3.37±0.30μM, comparable to that of positive control AMD (1.12±0.09μM), compounds11h,13d,13i showed anticancer activities against MGC-803with the IC50of2.02±0.23,4.88±0.28,2.96±0.15μM respectively, comparable to that of positive control AMD (1.01±0.08μM). According to preliminary anticancer activity of structure-activity relationship, for the acetyl-(sulfone)2H-pyrazole-coumarin skeleton, the substituent of5-aryl does not significantly change the activity; and the change with the acyl and sulfone of the N can greatly effect the inhibitory of these three kinds of cells; for acetyl-2H-pyrazole-coumarin skeleton compounds, the straight-chain amino of the1-N acetyl more favorable than the ring amine activity. The selected compounds were assayed for telomerase inhibition by a modified TRAP assay, the results show that the compounds11d,13a,13f with high anticancer activity of PC3, Bcap-37cells showed high activity against telomerase with the IC50of4.01±1.00,3.87±0.61,3.00±0.16μM respectively, comparable to that of positive control Ethidium bromide (2.63±0.7μM). For the structure of the cytotoxic drug activity and telomerase activity appear consistent, some ideas of the structure-based screening of telomerase inhibitors were provided. |
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