NET638250UC, Perkin Elmer), in 25 mM Tris-HCl buffer (pH 7.4) supplemented with 0.5 mM EDTA. process and all compounds were evaluated in vitro towards AChE and 5-HTRs. A docking study was performed in order to better clarify the observed SAR towards AChE, 5-HT4R and 5-HT6R and this study led to the description of novel ligand focusing on both AChE and 5-HT6R. = 3)(nM) = 3)(nM) (= 3)ideals are indicated as imply standard error of the imply (SEM) of three experiments; ND: not identified. As already introduced, we shown that pharmacomodulation of the preclinical candidate donecopride could greatly affect the ability of its analog to interact with ChE and 5-HT receptors. Indeed, in our initial study [6], we showed that its ester or amide analogs shed their ability to inhibit AChE. The same inclination has also been observed in the benzisoxazole family with a obvious impact of the modulation of the benzenic position on both AChE and 5-HT4R binding. [20] With this context, we decided to explore the modulation of the -position of the ketone and the possibility to generate conformationally constrained derivatives to increase the elucidation of SAR around donecopride. The preparation of first series of phenylpyrazoles 3aCb and 6, was then accomplished from your advance intermediate 1. With this synthesis, the safety of the basic nitrogen having a Boc group is definitely mandatory to obtain the pyrazole cycle, since all efforts to modulate the final donecopride analog inside a late stage process failed. For this reason, we also explored in parallel the effect of the safety of the aniline moiety with an acetamide group, with no obvious benefit in terms of synthesis of compound 6. The preparation of the Boc safeguarded 2 was, however, of interest since it allowed us to expose inside a convergent way both the cyclohexyl and the = 13.0 Hz, 2H), 2.29 (d, = 7.0 Hz, 2H), 1.54 (d, = 13.1 Hz, 2H), 1.45?1.38 (m, 10H), 0.97 (qd, = 12.2 Hz, = 4.2 Hz, 2H); 13C-NMR (CDCl3, 100 MHz): 156.8, 154.9, 147.4, 143.5, 131.7, 129.3, 118.7, 114.4, 110.7, 98.7, 79.2, 55.6, 44.0 (2C), 38.9, 37.0, 32.0 (2C), 31.1, (S)-(+)-Flurbiprofen 28.5 (3C); MS [M + H]+ = 434.86; IR (KBr, cm?1): 3468, 3351, 2931, 1686, 1624, 1461, 1444, 1426, 1365, 1247, 1211, 1173, 1211. General Procedure for Acetylation The aniline was stirred in anhydric acetic (6 mLmmol?1) at room temperature over night. The combination was concentrated under reduced pressure. The (S)-(+)-Flurbiprofen residue i had been then dissolved in EtOAc and washed twice with aq. NaHCO3 saturated and once with water. The organic phase was dried, filtrated, and evaporated in vacuo. The crude product is definitely purified on silica gel. = 12.8 Hz, 2H), 2.31 (d, = (S)-(+)-Flurbiprofen 7.0 Hz, 2H), 2.26 (s, 3H), 1.53 (d, = 13.2 Hz, 2H), 1.45?1.38 (m, 10H), 0.96 (qd, = 12.2 Hz, = 4.3 Hz, 2H); 13C-NMR (CDCl3, 100 MHz): 168.4, 155.7, 154.8, 154.4, 144.7, 134.4, 127.4, 119.3, 113.8, 104.4, 86.0, 79.5, 75.7, 55.9, 43.5 (2C), 36.1, 33.7, 28.6 (2C), 28.5 (3C), 25.1; MS [M + H]+ = 476.82; IR (KBr, (S)-(+)-Flurbiprofen cm?1): 3420, 2930, 1688, 1584, 1527, 1450, 1388, 1244, 1161. = 6.3 Hz, 2H), 3.89 (s, 3H), 3.66 (s, 3H), 2.73 (t, = 13.0 Hz, 2H), 2.23 Rabbit polyclonal to ACAP3 (s, 3H), 1.98 (m, 1H), 1.78 (d, = 12.6 Hz, 2H), 1.45 (s, 9H), 1.27 (qd, = 12.1 Hz, = 4.2 Hz, 2H); 13C-NMR (CDCl3, 100 MHz): 168.4, 155.7, 154.8, 154.4, 144.7, 134.4, 127.4, 119.3, 113.8, 104.4, 86.0, 79.5, 75.7, 55.9, 43.5 (2C), 36.1, 33.7, 28.6 (2C), 28.5 (3C), 25.1; [M + H]+ = 492.76; IR (KBr, cm?1): 3420, 2936, 2853, 1690, 1585, 1557, 1528, 1450, 1436, 1365, 1247, 1173, 1147, 1018. = 6.4 Hz, 2H), 3.68 (s, 3H), 2.75 (t, = 12.8 Hz, 2H), 2.00 (m, 1H), 1.80 (d, = 13.0 Hz, 2H), 1.47 (s, 9H), 1.29 (qd, = 11.8 Hz, = 4.6 Hz, 2H); 13C-NMR (CDCl3, 400 MHz): 155.1, 154.8, 149.3, 133.8, 128.5 (2C), 127.6, 125.2 (2C), 82.0, 79.6, 75.9, 43.5 (2C), 36.0, 33.8, 28.6 (2C), 28.5 (3C); MS [M + H]+ = 372.01; IR (KBr, cm?1): 3426, 2925, 2853, 1742, 1690, 1626, 1558, 1512, 1454, 1422, 1367, 1275. = 2.0 Hz, 1H), 7.22 (d, = 8.2 Hz, = 2.0 Hz, 1H), 6.86 (d, = 8.3 Hz, 1H), 5.74 (s, 1H), 4.18 (br s, 2H), 3.95 (s, 3H), 3.93 (d, = 6.4 Hz, 2H), 3.90 (s, 3H), 3.67 (s, 3H), 2.75 (t, = 13.0 Hz, 2H), 2.00 (m, 1H), 1.79 (d, = 10.3 Hz, 2H), 1.47 (s, 9H), 1.29 (qd, = 11.7 Hz, = 3.9 Hz, 2H); MS [M + H]+ = 431.91; General.