Table 3 Biological activities of isolated Vinca minor alkaloids.CompoundIC50 h AChE ± SEM (μM) aIC50 h BuChE ± SEM (μM) aIC50 POP ± SEM (μM) a% inhibition of GSK-3β SEM b ±logBB cPAMPA (P e; 10 6 cm s 1)dvincaminorine (1)> 100> 100429 ± 30–n.c.–vincaminoreine (2)> 1008.71 ± 0.49408 ± 60–0.262–eburnamonine (3)> 10093.49 ± 15.90n.d.–n.c.–minovine (4)> 10026.32 ± 2.52n.d.–n.c.–16-methoxyminovine (5)> 10025.01 ± 3.27n.t.–n.c.–vincatine (6)> 10043.47 ± 3.30n.d.–n.c.–minovincine (7)> 100> 100301 ± 15–n.c.–16-methoxyminovincine (8)> 100> 100n.d.–n.c.–vincaminorudeine (9)> 10084.91 ± 2.98n.d.–n.c.–demethoxyalstonamide (10)> 10056.38 ± 2.58n.d.–n.c.–vincorine (11)> 1009.75 ± 0.45n.t.–0.022–minovincinine (12)> 100> 100n.d.–n.c.–aspidospermidine (13)> 10033.60 ± 1.52n.d.–n.c.–19-oxoeburnamonine (14)> 100> 100n.t.–n.c.–akuammicine (15)> 10038.17 ± 0.84n.t.–n.c.–tubotaiwine (16)> 10011.70 ± 0.11n.d.–n.c.–raucubainine (17)> 10093.96 ± 6.80> 500–n.c.–aspidofractinine (18)> 100> 100n.d.–n.c.–2-ethyl-3[2-(3-ethylpiperidinyl)-ethyl]-1 H -> 1000.650 ± 0.01658 ± 431.43 ± 3.05¡0.16415.7 ± 1.3 (CNSþ)indole (19)dihydrovindolinine (20)> 100> 100n.d.–n.c.–strictamine (21)> 100> 100190 ± 11–n.c.–5-oxoaspidofractinine (22)> 100> 100n.t.–n.c.–raucubaine (23)> 100> 100n.t.–n.c.–galanthamine e1.72 ± 0.1242 ± 1––0.0475.1 (CNS) f +eserine e0.063 ± 0.0050.13 ± 0.01––0.177–berberine e0.72 ± 0.1131 ± 4142 ± 21–0.420–chlorothiazide e––––0.4181.1 ± 0.5 (CNS)a Compound concentration required to decrease enzyme activity by 50%; the values are the mean SEM of three independent measurements, each performed in ± triplicate.b Measured at a concentration of 100 μM.c calculated at http://www.way2drug.com/geb.d CNS(): high BBB permeation predicted with P (10 6 cm s 1)> 4.0, CNS(): low BBB permeation predicted with P (10 6 cm s 1) <2.0, CNS(): BBB permeation + e e ± uncertain with P (10 6 cm s 1) from 4.0 to 2.0.e e Reference compound; f Taken from reference (Stavrakov et al., 2017); n.t. stands for not tested due to the isolation of low amount; n.d. stands for not measurable due to a problem with solubility; n.c. stands for not calculated.

Table 3 Biological activities of isolated Vinca minor alkaloids.CompoundIC50 h AChE ± SEM (μM) aIC50 h BuChE ± SEM (μM) aIC50 POP ± SEM (μM) a% inhibition of GSK-3β SEM b ±logBB cPAMPA (P e; 10 6 cm s 1)dvincaminorine (1)> 100> 100429 ± 30–n.c.–vincaminoreine (2)> 1008.71 ± 0.49408 ± 60–0.262–eburnamonine (3)> 10093.49 ± 15.90n.d.–n.c.–minovine (4)> 10026.32 ± 2.52n.d.–n.c.–16-methoxyminovine (5)> 10025.01 ± 3.27n.t.–n.c.–vincatine (6)> 10043.47 ± 3.30n.d.–n.c.–minovincine (7)> 100> 100301 ± 15–n.c.–16-methoxyminovincine (8)> 100> 100n.d.–n.c.–vincaminorudeine (9)> 10084.91 ± 2.98n.d.–n.c.–demethoxyalstonamide (10)> 10056.38 ± 2.58n.d.–n.c.–vincorine (11)> 1009.75 ± 0.45n.t.–0.022–minovincinine (12)> 100> 100n.d.–n.c.–aspidospermidine (13)> 10033.60 ± 1.52n.d.–n.c.–19-oxoeburnamonine (14)> 100> 100n.t.–n.c.–akuammicine (15)> 10038.17 ± 0.84n.t.–n.c.–tubotaiwine (16)> 10011.70 ± 0.11n.d.–n.c.–raucubainine (17)> 10093.96 ± 6.80> 500–n.c.–aspidofractinine (18)> 100> 100n.d.–n.c.–2-ethyl-3[2-(3-ethylpiperidinyl)-ethyl]-1 H -> 1000.650 ± 0.01658 ± 431.43 ± 3.05¡0.16415.7 ± 1.3 (CNSþ)indole (19)dihydrovindolinine (20)> 100> 100n.d.–n.c.–strictamine (21)> 100> 100190 ± 11–n.c.–5-oxoaspidofractinine (22)> 100> 100n.t.–n.c.–raucubaine (23)> 100> 100n.t.–n.c.–galanthamine e1.72 ± 0.1242 ± 1––0.0475.1 (CNS) f +eserine e0.063 ± 0.0050.13 ± 0.01––0.177–berberine e0.72 ± 0.1131 ± 4142 ± 21–0.420–chlorothiazide e––––0.4181.1 ± 0.5 (CNS)a Compound concentration required to decrease enzyme activity by 50%; the values are the mean SEM of three independent measurements, each performed in ± triplicate.b Measured at a concentration of 100 μM.c calculated at http://www.way2drug.com/geb.d CNS(): high BBB permeation predicted with P (10 6 cm s 1)> 4.0, CNS(): low BBB permeation predicted with P (10 6 cm s 1) <2.0, CNS(): BBB permeation + e e ± uncertain with P (10 6 cm s 1) from 4.0 to 2.0.e e Reference compound; f Taken from reference (Stavrakov et al., 2017); n.t. stands for not tested due to the isolation of low amount; n.d. stands for not measurable due to a problem with solubility; n.c. stands for not calculated.

Table 1 Key correlations from 2D NMR experiments in the structure elucidation of undescribed alkaloid 9 (HMBC interaction – blue arrow; H2BC and COSY interaction – red bond) and its 1 H and 13 C NMR chemical shifts assigned to the related position in ppm with coupling constants (in parenthesis, reported in Hz)..Positionvincaminorudeine 9500 MHz, CDCl3125 MHz, CDCl32139.134.39, dd (4.1, 2.7)38.742.63–2.55, m31.11.92, dd (14.3, 4.1)542.961.36–1.21, m31.71.06, td (12.5, 5.5)71.52–1.39, m22.31.36–1.21, m82.38–2.22, m55.2102.63–2.55, m53.62.38–2.22, m113.05–2.85, m22.312110.313127.0147.53, dd (7.8, 1.1)118.1157.09, dd (7.8, 1.1)118.5167.19, dd (7.8, 1.1)120.7177.27, dd (7.8, 1.1)108.618136.7193.75–3.70, m, overlap56.61.64, d (12.5)203.47, q (6.5)74.2211.29, d, overlap (6.5)17.6C= O177.03-COOCH33.72, s, overlap52.7N CH33.54, s30.3

Table 1 Key correlations from 2D NMR experiments in the structure elucidation of undescribed alkaloid 9 (HMBC interaction – blue arrow; H2BC and COSY interaction – red bond) and its 1 H and 13 C NMR chemical shifts assigned to the related position in ppm with coupling constants (in parenthesis, reported in Hz)..Positionvincaminorudeine 9500 MHz, CDCl3125 MHz, CDCl32139.134.39, dd (4.1, 2.7)38.742.63–2.55, m31.11.92, dd (14.3, 4.1)542.961.36–1.21, m31.71.06, td (12.5, 5.5)71.52–1.39, m22.31.36–1.21, m82.38–2.22, m55.2102.63–2.55, m53.62.38–2.22, m113.05–2.85, m22.312110.313127.0147.53, dd (7.8, 1.1)118.1157.09, dd (7.8, 1.1)118.5167.19, dd (7.8, 1.1)120.7177.27, dd (7.8, 1.1)108.618136.7193.75–3.70, m, overlap56.61.64, d (12.5)203.47, q (6.5)74.2211.29, d, overlap (6.5)17.6C= O177.03-COOCH33.72, s, overlap52.7N CH33.54, s30.3

Table 2 Comparison of experimental NMR data of this work with those presented by Farahanikia et al. (2011) for the very same structure of vincaminoreine 2. The specific optical rotation was consistent with the published one by Farahanikia et al. (2011) and Mokrý et al. (1964). 1 H and 13 C NMR chemical shifts are assigned to the related position in ppm with coupling constants for 1 H NMR (in parenthesis, reported in Hz)..PositionExperimental vincaminoreine 2published vincaminoreine 2 (Farahanikia et al., 2011)500 MHz, CDCl3125 MHz, CDCl3500 MHz, CDCl3125 MHz, CDCl32139.3139.333.90, dd (4.8, 2.9)38.11.96–1.93, m33.842.46, dd (13.7, 2.9)33.92.95–2.93, m22.51.97, d (13.7)538.124.861.43–1.36, m34.21.35–1.30, m31.01.24–1.12, m1.20–1.18, m71.54–1.44, m22.51.54–1.47, m22.31.35–1.24, m1.29–1.25, m82.38–2.32, m55.32.51–2.49, m53.32.32–2.20, m2.27–2.24, m102.55–2.50, m53.32.33–2.29, m55.32.32–2.20, m2.29–2.27, m113.00–2.89, m22.31.41–1.36, m34.21.18–1.16, m12110.0110.013127.1127.1147.50, dd (7.8, 1.0)118.07.50, d (7.1)118.0157.18, td (7.8, 1.0)118.57.17, t (7.0)120.6167.09, td (7.8, 1.0)120.67.07, t (7.0)118.5177.26, dd (7.8, 1.0)108.67.24, overlap108.618136.8136.7193.16, d (12.5)57.93.16, d (15.0)57.91.61, d (12.5)1.61, d (15.0)201.35–1.24, m31.01.25–1.20, m14.21.24–1.12, m210.95, t (7.5)7.40.93, t (7.2)7.4C= O175.2175.23-COOCH33.71, s52.53.70, s52.5NCH33.54, s30.23.52, s30.2[a]D+ 27.6 (c 0.18, CHCl3)+ 26.5 (CHCl3)(Farahanikia et al., 2011; Mokrý et al., 1964)

Table 2 Comparison of experimental NMR data of this work with those presented by Farahanikia et al. (2011) for the very same structure of vincaminoreine 2. The specific optical rotation was consistent with the published one by Farahanikia et al. (2011) and Mokrý et al. (1964). 1 H and 13 C NMR chemical shifts are assigned to the related position in ppm with coupling constants for 1 H NMR (in parenthesis, reported in Hz)..PositionExperimental vincaminoreine 2published vincaminoreine 2 (Farahanikia et al., 2011)500 MHz, CDCl3125 MHz, CDCl3500 MHz, CDCl3125 MHz, CDCl32139.3139.333.90, dd (4.8, 2.9)38.11.96–1.93, m33.842.46, dd (13.7, 2.9)33.92.95–2.93, m22.51.97, d (13.7)538.124.861.43–1.36, m34.21.35–1.30, m31.01.24–1.12, m1.20–1.18, m71.54–1.44, m22.51.54–1.47, m22.31.35–1.24, m1.29–1.25, m82.38–2.32, m55.32.51–2.49, m53.32.32–2.20, m2.27–2.24, m102.55–2.50, m53.32.33–2.29, m55.32.32–2.20, m2.29–2.27, m113.00–2.89, m22.31.41–1.36, m34.21.18–1.16, m12110.0110.013127.1127.1147.50, dd (7.8, 1.0)118.07.50, d (7.1)118.0157.18, td (7.8, 1.0)118.57.17, t (7.0)120.6167.09, td (7.8, 1.0)120.67.07, t (7.0)118.5177.26, dd (7.8, 1.0)108.67.24, overlap108.618136.8136.7193.16, d (12.5)57.93.16, d (15.0)57.91.61, d (12.5)1.61, d (15.0)201.35–1.24, m31.01.25–1.20, m14.21.24–1.12, m210.95, t (7.5)7.40.93, t (7.2)7.4C= O175.2175.23-COOCH33.71, s52.53.70, s52.5NCH33.54, s30.23.52, s30.2[a]D+ 27.6 (c 0.18, CHCl3)+ 26.5 (CHCl3)(Farahanikia et al., 2011; Mokrý et al., 1964)

This dataset contains the digitized treatments in Plazi based on the original journal article Vrabec, Rudolf, Ma, Jana, ríkov, a, Loc, Miroslav, arek, Kor, Jan, abe, cný, Hulcova, Daniela, Ho, Anna, st, alkov, a, Ji, Kune, rí, s, Chlebek, Jakub, Toma, Ku, s, cera, Hrabinova, Martina, Jun, Daniel, Ond, Soukup, rej, Andrisano, Vincenza, Jen, Jaroslav, co, Marcela, Safratova, Nov, Lucie, akova, Opletal, Lubomír, Cahlíkova, Lucie (2022): Monoterpene indole alkaloids from Vinca minor L. (Apocynaceae): Identification of new structural scaffold for treatment of Alzheimer’s disease. Phytochemistry (113017) 194: 1-13, DOI: 10.1016/j.phytochem.2021.113017, URL: http://dx.doi.org/10.1016/j.phytochem.2021.113017

Table 1 1 H-NMR and 13 C-NMR data of new alkaloid 6 (diastereomeric ratio 1:1.1) isolated from Narcissus pseudonarcissus cv. Duch Master.POSITIONNARCIMATULINEδC ofδH of isomer AδC ofδH of isomer Bisomer Aisomer B129.72.39–2.26 m29.62.39–2.26 m1.84–1.64 m1.84–1.64 m235.72.39–2.26 m35.62.39–2.26 m2.13–1.98 m2.13–1.98 m3209.2209.2440.02.60 dd, overlapped40.02.56 dd, overlapped(18.4, 2.9)(18.0, 2.9)4a88.34.68 t (2.9)88.24.55 t (2.9)5a146.7146.76143.6143.67110.96.65 d, overlapped111.06.62 d, overlapped(8.0)(8.0)8122.236.44 d (8.0)122.206.56 d (8.0)8a129.9129.98b132.0132.0957.63.88 d, overlapped57.93.98 d (15.0)(15.0)3.66 d (15.0)3.51 d (15.0)1152.33.05–2.88 m51.13.05–2.88 m2.86–2.74 m1237.42.13–1.98 m36.41.84–1.64 m1.56–1.50 m1.38–1.31 m1347.547.41′54.93.45 d (14.2)53.73.35 bs3.33–3.24 m2′122.8122.83′108.77.01 s108.47.07 s3a′146.9146.95′101.06.00–5.98 m100.96.00–5.98 m6a′146.0145.87ֹ’110.56.74 s110.36.77 s8′133.6133.49′131.7131.410′130.46.83 d (7.4)130.26.71 d, overlapped(7.4)11′117.96.68 t, overlapped117.96.62 t, overlapped(7.4)(7.4)12′123.227.03 d, overlapped123.247.03 d, overlapped(7.4)(7.4)12a′131.0131.013′28.63.05–2.88 m28.53.05–2.88 m2.86–2.74 m2.86–2.74 m14′57.13.33–3.24 m56.93.33–3.24 m3.18 dd (17.5, 8.8)3.09 dd (18.4, 9.2)15a′150.5150.4N15′-CH338.82.25 s38.72.20 sC6-OCH355.963.86–3.83 m55.953.86–3.83 mSpectra were acquired in chloroform-d at 500 MHz (1 H) and 125.7 MHz (13 C);1 J values are in parentheses and reported in Hz; chemical shifts are given in ppm.

Table 1 1 H-NMR and 13 C-NMR data of new alkaloid 6 (diastereomeric ratio 1:1.1) isolated from Narcissus pseudonarcissus cv. Duch Master.POSITIONNARCIMATULINEδC ofδH of isomer AδC ofδH of isomer Bisomer Aisomer B129.72.39–2.26 m29.62.39–2.26 m1.84–1.64 m1.84–1.64 m235.72.39–2.26 m35.62.39–2.26 m2.13–1.98 m2.13–1.98 m3209.2209.2440.02.60 dd, overlapped40.02.56 dd, overlapped(18.4, 2.9)(18.0, 2.9)4a88.34.68 t (2.9)88.24.55 t (2.9)5a146.7146.76143.6143.67110.96.65 d, overlapped111.06.62 d, overlapped(8.0)(8.0)8122.236.44 d (8.0)122.206.56 d (8.0)8a129.9129.98b132.0132.0957.63.88 d, overlapped57.93.98 d (15.0)(15.0)3.66 d (15.0)3.51 d (15.0)1152.33.05–2.88 m51.13.05–2.88 m2.86–2.74 m1237.42.13–1.98 m36.41.84–1.64 m1.56–1.50 m1.38–1.31 m1347.547.41′54.93.45 d (14.2)53.73.35 bs3.33–3.24 m2′122.8122.83′108.77.01 s108.47.07 s3a′146.9146.95′101.06.00–5.98 m100.96.00–5.98 m6a′146.0145.87ֹ’110.56.74 s110.36.77 s8′133.6133.49′131.7131.410′130.46.83 d (7.4)130.26.71 d, overlapped(7.4)11′117.96.68 t, overlapped117.96.62 t, overlapped(7.4)(7.4)12′123.227.03 d, overlapped123.247.03 d, overlapped(7.4)(7.4)12a′131.0131.013′28.63.05–2.88 m28.53.05–2.88 m2.86–2.74 m2.86–2.74 m14′57.13.33–3.24 m56.93.33–3.24 m3.18 dd (17.5, 8.8)3.09 dd (18.4, 9.2)15a′150.5150.4N15′-CH338.82.25 s38.72.20 sC6-OCH355.963.86–3.83 m55.953.86–3.83 mSpectra were acquired in chloroform-d at 500 MHz (1 H) and 125.7 MHz (13 C);1 J values are in parentheses and reported in Hz; chemical shifts are given in ppm.

Table 2 h AChE, h BuChE, POP and GSK-3β inhibitory activities of the tested Amaryllidaceae alkaloids isolated from Narcissus pseudonarcissus cv. Dutch Master expressed as IC50.Compoundh AChE IC (μM) a 50h BuChE IC (μM) a 50SI for h AChE bPOP IC a (μM) 50GSK-3β (% inhib.) cGSK-3β IC a (μM) 50Homolycorine (1)64 ± 4151 ± 202.4173 ± 4154 ± 1n.d.Masonine (2)305 ± 34229 ± 240.8314 ± 3466 ± 427.9 ± 0.8Seco-isopowellaminone (3)294 ± 33> 500> 1.7> 50038 ± 1n.d.O -Ethyllycorenine (4)> 500> 500n.d.> 50058 ± 3n.d.N -Demethylmasonine (5)> 500474 ± 13<0.9> 500n.d.n.d.Narcimatuline (6)489 ± 605.9 ± 0.20.0129.2 ± 1.067 ± 320.7 ± 2.4O -Acetylpluviine (7)> 500> 500n.d.n.d.n.d.n.d.Tazettine (8)> 500> 500n.d.> 50049 ± 0n.d.Galanthine (9)606 ± 60> 500> 1.7> 50026 ± 7n.d.Galanthamine (10)1.7 ± 0.142.3 ± 1.324.9> 500n.d.n.d.Narwedine (11)281 ± 34911 ± 693.2907 ± 87n.d.n.d.Caranine (12)321 ± 42487 ± 551.5> 100058 ± 930.8 ± 0.3Lycoraminone (13)> 500> 500n.d.n.d.39 ± 1n.d.Lycoramine (14)456 ± 57> 500> 1.1> 500n.d.n.d.O -Methyllycorenine (15)> 500> 500n.d.> 500n.d.n.d.Lycorenine (16)> 500> 500n.d.> 50048 ± 3n.d.Oduline (17)> 500> 500n.d.252 ± 2758 ± 4n.d.Haemanthamine (18)> 500> 500n.d.> 50052 ± 0n.d.Tetrahydromasonine (19)> 500> 500n.d.> 50022 ± 0n.d.Hippeastrine (20)> 500> 500n.d.> 50011 ± 2n.d.Crinine (21)> 500> 500n.d.> 50040 ± 5n.d.Epimaritidine (22)> 500> 500n.d.n.d.n.d.n.d.Huperzine A c0.033 ± 0.001> 500> 15.151n.d.n.d.n.d.Berberine cn.d.n.d.n.d.142 ± 21n.d.n.d.Z -Pro-prolinal cn.d.n.d.n.d.2.75 × 10−3n.d.n.d.SB-415286 cn.d.n.d.n.d.n.d.n.d.70 nMa Compound concentration required to decrease enzyme activity by 50%; the values are the mean ± SEM of three independent measurements, each performed in triplicate; b Selectivity index (SI) for h AChE is determined as ratio h BuChE IC / h AChE IC; c tested at 10 μM compound concentration; n.d. stands for not determined.