Molecules 2016, 21, 132
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2-(4-((3-(Dimethylamino)propylamino)methyl)phenyl)-1-methylquinolin-4(1H)-one (5a). A light-yellow solid.
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m.p. 182–185 C; H-NMR (400 MHz, CDCl3):
δ 8.17 (dd, J = 7.6, 1.4 Hz, 1H), 7.95–7.64 (m, 3H),
7.46 (d, J = 7.6 Hz, 2H), 7.22 (td, J = 7.6, 1.6 Hz, 1H), 7.08 (dd, J = 7.6, 1.4 Hz, 1H), 7.01 (s, 1H),
6.50 (s, 1H), 3.92 (s, 2H), 3.54 (s, 3H), 2.61 (t, J = 7.6 Hz, 2H), 2.50 (t, J = 7.6 Hz, 2H), 2.27 (s, 6H),
1.42 (m, 2H); APCI-MS m/z: 350.2 [M + H]+. Calcd. for C22H27N3O. H2O: C, 71.90; H, 7.95; N, 11.43.
Found: C, 72.03; H, 7.94; N, 11.37.
1-Methyl-2-(4-((3-(pyrrolidin-1-yl)propylamino)methyl)phenyl)quinolin-4(1H)-one (5b). A light-yellow solid.
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m.p. 176–180 C; H-NMR (400 MHz, CDCl3):
δ 8.31 (dd, J = 7.6, 1.4 Hz, 1H), 7.89–7.64 (m, 3H),
7.48 (d, J = 7.6 Hz, 2H), 7.32 (td, J = 7.6, 1.6 Hz, 1H), 6.99 (dd, J = 7.6, 1.4 Hz, 1H), 6.88 (s, 1H),
6.45 (s, 1H), 3.94 (s, 2H), 3.52 (s, 3H), 3.07 (t, 2H), 2.62 (t, J = 7.6 Hz, 2H), 2.48 (t, 2H), 2.32 (m, 4H),
1.66 (m, 4H); APCI-MS m/z: 376.2 [M + H]+. Calcd. for C24H29N3O. 2H2O: C, 70.04; H, 8.08; N, 10.21.
Found: C, 70.09; H, 7.99; N, 10.34.
1-Methyl-2-(4-((3-(piperidin-1-yl)propylamino)methyl)phenyl)quinolin-4(1H)-one (5c). A light-yellow solid.
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m.p. 177–179 C; H-NMR (400 MHz, CDCl3):
δ 8.26 (dd, J = 7.4, 1.4 Hz, 1H), 7.65 (ddd, J = 10.8,
8.2, 4.4 Hz, 3H), 7.32 (d, J = 7.5 Hz, 2H), 7.28 (td, J = 7.6, 1.6 Hz, 1H), 7.16 (dd, J = 7.6, 1.4 Hz, 1H),
6.88 (s, 1H), 6.45 (s, 1H), 3.97 (s, 2H), 3.53 (s, 3H), 3.14 (t, J = 5.4 Hz, 2H), 2.60 (dd, J = 16.0, 7.8 Hz, 2H),
2.37 (dt, J = 10.7, 6.5 Hz, 4H), 1.67–1.60 (m, 6H), 1.52 (m, 2H); APCI-MS m/z: 390.2 [M + H]+. Calcd. for
C25H31N3O. H2O: C, 73.68; H, 8.16; N, 10.31. Found: C, 73.54; H, 8.27; N, 10.26.
2-(4-((4-(Dimethylamino)butylamino)methyl)phenyl)-1-methylquinolin-4(1H)-one (6a). A light-yellow
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solid. m.p. 173–176 C; H-NMR (400 MHz, CDCl3):
δ 8.25 (dd, J = 7.6, 1.4 Hz, 1H), 7.68–7.55
(m, 3H), 7.32 (d, J = 7.6 Hz, 2H), 7.17 (td, J = 7.6, 1.4 Hz, 1H), 6.99 (dd, J = 7.4, 1.5 Hz, 1H),
6.85 (s, 1H), 6.50 (s, 1H), 3.95 (s, 2H), 3.54 (s, 3H), 2.73 (t, 2H), 2.62 (t, 2H), 2.39 (s, 6H), 1.76 (m, 2H),
1.63 (m, 2H); APCI-MS m/z: 364.2 [M + H]+. Calcd. for C23H29N3O. 2H2O: C, 69.14; H, 8.33; N, 10.52.
Found: C, 69.34; H, 8.39; N, 10.47.
1-Methyl-2-(4-((4-(pyrrolidin-1-yl)butylamino)methyl)phenyl)quinolin-4(1H)-one (6b). A light-yellow solid.
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m.p. 169–172 C; H-NMR (400 MHz, CDCl3):
δ 8.21 (dd, J = 7.6, 1.4 Hz, 1H), 7.68–7.55 (m, 3H),
7.44 (d, J = 7.6 Hz, 2H), 7.23 (td, J = 7.6, 1.6 Hz, 1H), 7.14 (dd, J = 7.6, 1.4 Hz, 1H), 7.02 (s, 1H),
6.45 (s, 1H), 3.98 (s, 2H), 3.50 (s, 3H), 3.13 (t, 2H), 2.68 (t, 2H), 2.40 (t, 2H), 2.14 (m, 4H), 1.98–1.31
(m, 6H); APCI-MS m/z: 390.3 [M + H]+. Calcd. for C25H31N3O. 2H2O: C, 70.56; H, 8.29; N, 9.87. Found:
C, 70.44; H, 8.31; N, 9.93.
3.2. Materials
Stock solutions of all graveoline analogs (10 mM) were prepared using DMSO and stored at
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80 C. Further dilutions to working concentrations were made with double-distilled deionized
water. AChE (E.C. 3.1.1.7, obtained from electric eel), BuChE (E.C. 3.1.1.8, obtained from equine
serum), 5,51-dithiobis-(2-nitrobenzoic acid) (Ellman’s reagent, DTNB), acetylthiocholine chloride
(ATC), butylthiocholine chloride, thioflavin T, and tarcine hydrochloride were purchased from
Sigma-Aldrich (St. Louis, MO, USA). Aβ42 was purchased from Millipore (Billerica, MA, USA).
3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was purchased formal Aladdin
(Shanghai, China). The SH-SY5Y cell line was obtained from the Animal experimental center of
Sun Yat-sen University.
3.3. In Vitro Inhibition of AChE and BuChE
In vitro AChE assay: all methods were performed under 0.1 M KH2PO4/K2HPO4 buffer at pH
8.0 by using a Shimadzu 2450 spectrophotometer. The solutions of the enzyme were prepared to
achieve 2.0 units/mL in 2 mL aliquots. The experiment system contained phosphate buffer, pH 8.0
(1 mL), 50
µL of 0.01 M substrate (ATC), 50 µL of 0.01 M DTNB, and 10 µL of enzyme. The substrate
was added into the experiment system containing the DTNB, buffer, and enzyme with inhibitors