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Compd.	Yield /%	mp/℃	$ {\left[\alpha \right]}_{\mathrm{D}}^{25\ ^\circ {\rm{C}}} $ (g·100 mL^-1, MeOH)	¹H NMR (600 MHz)	¹³C NMR (151 MHz)	HPLC purity	HR-ESI-MS (m/z)
5a	28%	51-53	-10.00 (0.030)	(CDCl₃) δ 6.86 (s, 1H), 6.59 (s, 1H), 5.96-5.88 (m, 2H), 5.56 (s, 1H), 4.58 (s, 1H), 4.11 (d, J = 14.0 Hz, 1H), 3.92-3.88 (m, 1H), 3.67-3.60 (m, 2H), 3.50 (d, J = 14.0 Hz, 1H), 3.33-3.30 (m, 1H), 2.71 (s, 2H), 2.66-2.57 (m, 2H), 2.36 (q, J = 8.7 Hz, 1H), 1.65-1.52 (m, 2H), 1.45-1.32 (m, 2H), 0.93 (t, J = 7.4 Hz, 3H).	(CDCl₃) δ 146.7, 146.3, 143.5, 130.6, 127.7, 116.0, 107.9, 104.6, 101.0, 79.4, 69.9, 69.0, 61.0, 57.1, 53.9, 41.8, 32.2, 28.8, 19.4, 14.0.	96.9%	[M+H]⁺: C₂₀H₂₆O₄N 344.185 6 344.184 0
5b	20%	114-116	40.91 (0.022)	(CDCl₃) δ 6.86 (s, 1H), 6.60 (s, 1H), 5.96-5.91 (m, 2H), 5.56 (s, 1H), 4.58 (s, 1H), 4.12 (d, J = 14.1 Hz, 1H), 3.92 (s, 1H), 3.72-3.63 (m, 2H), 3.60-3.47 (m, 4H), 3.35-3.32 (m, 1H), 2.71 (s, 2H), 2.66-2.57 (m, 2H), 1.91-1.84 (m, 2H), 1.78-1.73 (m, 2H).	(CDCl₃) δ 146.7, 146.4, 143.7, 130.6, 127.5, 115.8, 107.9, 104.6, 101.1, 79.4, 70.2, 69.1, 61.0, 57.1, 53.9, 45.0, 41.8, 29.5, 28.8, 27.5.	95.2%	[M+H]⁺: C₂₀H₂₅O₄NCl 378.146 7 378.145 2
5c	45%	64-66	-5.56 (0.018)	(DMSO-d₆) δ 6.87 (s, 1H), 6.66 (s, 1H), 5.94 (d, J = 5.3 Hz, 2H), 5.85-5.79 (m, 1H), 5.42-5.39 (m, 1H), 5.01 (dd, J = 17.2, 1.9 Hz, 1H), 4.95 (dd, J = 17.2, 1.9 Hz, 1H), 4.92-4.75 (m, 1H), 4.41 (s, 1H), 4.00 (d, J = 14.1 Hz, 1H), 3.73-3.70 (m, 1H), 3.62 (dt, J = 9.4, 6.2 Hz, 1H), 3.52 (dt, J = 9.4, 6.2 Hz, 1H), 3.43-3.25 (m, 2H), 3.21-3.14 (m, 1H), 2.59 (d, J = 10.4 Hz, 1H), 2.47-2.39 (m, 1H), 2.36 (d, J = 10.4 Hz, 1H), 2.20 (q, J = 8.6 Hz, 1H), 2.07 (q, J = 7.4 Hz, 2H), 1.64-1.53 (m, 2H).	(DMSO-d₆) δ 146.2, 145.7, 143.6, 138.9, 130.1, 129.6, 116.3, 115.4, 107.5, 105.6, 101.1, 80.3, 68.8, 66.9, 61.1, 57.0, 53.6, 41.3, 30.3, 29.2, 28.6.	98.8%	[M+H]⁺: C₂₁H₂₆O₄N 356.185 6 356.184 1
6c	30%	102-104	64.71 (0.017)	(CD₃OD) δ 7.03-6.98 (m, 2H), 5.99 (d, J = 1.2 Hz, 1H), 5.97 (d, J = 1.2 Hz, 1H) 5.74-5.70 (m, 1H), 5.68-5.56 (m, 1H), 5.14 (dt, J = 10.1, 1.4 Hz, 1H), 5.07 (dt, J = 10.1, 1.4 Hz, 1H), 4.77-4.75 (m, 1H), 4.55-4.52 (m, 1H), 4.26-4.15 (m, 3H), 3.92-3.83 (m, 1H), 3.56-3.51 (m, 1H), 3.38-3.33 (m, 1H), 3.08 (d, J = 2.4 Hz, 1H), 3.02-2.97 (m, 2H), 2.85-2.78 (m, 1H), 2.53-2.48 (m, 2H).	(CD₃OD) δ 150.8, 148.5, 137.5, 133.1, 131.6, 124.8, 123.5, 119.6, 109.7, 106.7, 103.1, 73.2, 71.6, 69.9, 64.9, 64.1, 62.6, 39.6, 31.0, 29.1.	98.0%	[M]⁺: C₂₀H₂₄O₄N 342.170 0 342.170 0
6d	20%	150-152	65.22 (0.023)	(CD₃OD) δ 7.00 (s, 1H), 6.98 (s, 1H), 5.99 (d, J = 1.2 Hz, 1H), 5.97 (d, J = 1.2 Hz, 1H), 5.75-5.67 (m, 2H), 5.04-4.87 (m, 2H), 4.73 (d, J = 13.8 Hz, 1H), 4.53-4.51 (m, 1H), 4.24-4.08 (m, 3H), 3.54-3.47 (m, 1H), 3.29 (s, 1H), 3.21-3.10 (m, 1H), 3.01-2.93 (m, 3H), 2.84-2.78 (m, 1H), 2.05-1.93 (m, 2H), 1.86-1.76 (m, 2H).	(CD₃OD) δ 150.8, 148.4, 137.5, 137.4, 131.6, 124.8, 123.4, 117.0, 109.8, 106.7, 103.1, 73.3, 71.5, 69.9, 65.5, 63.9, 62.4, 39.5, 31.2, 31.0, 23.8.	95.5%	[M]⁺: C₂₁H₂₆O₄N 356.185 6 356.184 4
6e	25%	250-252	69.57 (0.023)	(CD₃OD) δ 6.74-6.70 (m, 2H), 5.71-5.66 (m, 2H), 5.45 (d, J = 3.5 Hz, 1H), 4.50-4.44 (m, 2H), 4.27-4.22 (m, 1H), 3.97-3.84 (m, 3H), 2.97-2.92 (m, 4H), 2.77-2.67 (m, 3H), 2.58-2.51 (m, 1H), 1.68-1.53 (m, 2H), 1.46-1.34 (m, 2H).	(CD₃OD) δ 150.8, 148.5, 137.5, 131.6, 124.8, 123.4, 109.8, 106.7, 103.1, 73.3, 71.6, 69.9, 65.0, 64.0, 62.4, 44.6, 39.6, 31.0, 30.2, 22.2.	98.0%	[M]⁺: C₂₀H₂₅O₄NCl 378.146 7 378.144 9
6f	20%	170-172	7.50 (0.040)	(CD₃OD) δ 7.00 (s, 1H), 6.99 (s, 1H), 5.97 (d, J = 3.8 Hz, 2H), 5.74-5.70 (m, 2H), 4.91 (dd, J = 16.5, 2.2 Hz, 1H), 4.85 (dd, J = 10.5, 2.1 Hz, 1H), 4.77-4.72 (m, 3H), 4.52 (s, 2H), 4.23-4.12 (m, 3H), 3.53 (td, J = 12.8, 6.6 Hz, 1H), 3.17 (td, J = 12.4, 4.5 Hz, 1H), 3.01-2.94 (m, 3H), 2.83-2.77 (m, 1H), 1.98-1.95 (m, 2H), 1.75-1.65 (m, 2H), 1.26-1.18 (m, 10H).	(CD₃OD) δ 150.7, 148.4, 140.1, 137.6, 131.6, 124.9, 123.3, 114.7, 109.7, 106.7, 103.1, 73.1, 71.6, 69.9, 66.1, 63.9, 62.4, 39.5, 34.8, 31.0, 30.2, 30.1, 30.0 (2), 27.3, 24.5.	95.2%	[M]⁺: C₂₆H₃₆O₄N 426.263 9 426.262 3
6g	20%	213-215	66.67 (0.030)	(CD₃OD) δ 7.39-7.34 (m, 2H), 7.30 (d, J = 7.9 Hz, 2H), 7.11-6.95 (m, 2H), 6.00 (dd, J = 6.7, 1.3 Hz, 2H), 5.72-5.63 (m, 1H), 4.65-4.48 (m, 2H), 4.27 (d, J = 13.2 Hz, 1H), 4.22-4.09 (m, 5H), 3.42-3.35 (m, 1H), 2.98 (d, J = 11.4 Hz, 1H), 2.74-2.54 (m, 2H), 2.35 (s, 3H).	(CD₃OD) δ 150.8, 148.5, 142.8, 137.6, 134.1 (2), 131.9, 131.3 (2), 126.4, 124.6, 123.4, 110.2, 106.8, 103.2, 72.6, 71.4, 69.9, 68.3, 62.9, 62.4, 39.5, 30.9, 21.3.	96.2%	[M]⁺: C₂₄H₂₆O₄N 392.185 6 392.184 6
6h	50%	214-216	29.03 (0.031)	(CD₃OD) δ 7.84-7.80 (m, 2H), 7.25-7.21 (m, 2H), 7.01 (s, 1H), 6.95 (s, 1H), 6.01 (d, J = 1.2 Hz, 1H), 5.99 (d, J = 1.2 Hz, 1H), 5.72-5.69 (m, 1H), 4.66-4.59 (m, 1H), 4.55-4.53 (m, 1H), 4.31 (d, J = 13.3 Hz, 1H), 4.25 (d, J = 13.3 Hz, 1H), 4.20-4.14 (m, 4H), 3.47-3.40 (m, 1H), 2.97 (d, J = 11.7 Hz, 1H), 2.76-2.69 (m, 2H).	(CD₃OD) δ 150.7, 148.4, 139.9 (2), 137.5, 135.8 (2), 131.7, 129.1, 124.5, 123.6, 110.1, 106.8, 103.2, 98.5, 72.8, 71.4, 69.8, 67.9, 63.6, 62.6, 39.4, 30.8.	96.2%	[M]⁺: C₂₃H₂₃O₄NI 504.066 6 504.065 0
6i	32%	63-65	8.57 (0.035)	(CD₃OD) δ 7.47-7.38 (m, 2H), 7.25-7.11 (m, 2H), 6.81 (s, 1H), 6.56 (s, 1H), 5.89-5.79 (m, 2H), 5.56-5.53 (m, 1H), 4.73-4.64 (m, 2H), 4.59-4.56 (m, 1H), 4.06-4.02 (m, 2H), 3.94-3.92 (m, 1H), 3.49-3.43 (m, 1H), 2.93 (s, 1H), 2.83-2.77 (m, 2H), 2.67-2.57 (m, 2H), 2.56-2.50 (m, 1H), 2.36 (q, J = 8.9 Hz, 1H).	(CD₃OD) δ 148.1, 147.7, 144.7, 139.6, 130.4, 130.3 (2), 129.6, 122.8, 122.0 (2), 121.1, 117.3, 108.2, 106.1, 102.3, 80.9, 71.7, 68.9, 62.4, 57.8, 54.7, 42.2, 29.3.	97.5%	[M]⁺: C₂₄H₂₃O₅NF₃ 462.152 3 462.150 3
6j	44%	210-212	46.43 (0.028)	(CD₃OD) δ 7.68 (d, J = 1.7 Hz, 1H), 7.62 (d, J = 1.7 Hz, 2H), 6.98 (s, 1H), 6.70 (s, 1H), 6.00–5.94 (m, 2H), 5.75-5.71 (m, 1H), 4.69 (d, J = 14.1 Hz, 1H), 4.53-4.51 (m, 1H), 4.44-4.39 (m, 2H), 4.31-4.18 (m, 4H), 3.66-3.53 (m, 1H), 2.94 (d, J = 11.5 Hz, 1H), 2.82-2.78 (m, 2H).	(CD₃OD) δ 150.6, 148.3, 137.4, 135.8 (3), 135.5, 133.6, 131.5, 124.6, 123.7 (2), 109.6, 106.6, 103.2, 72.4, 71.4, 69.8, 67.7, 65.7, 63.3, 54.2, 30.8.	98.0%	[M]⁺: C₂₃H₂₂O₄NBr₂ 533.991 0 533.989 3 (⁷⁹Br+⁷⁹Br) 535.987 4 (⁷⁹Br+⁸¹Br) 537.985 2 (⁸¹Br+⁸¹Br)
9a	24%	194-196	17.39 (0.023)	(CD₃OD) δ 7.84-7.81 (m, 2H), 7.26-7.20 (m, 2H), 6.93 (s, 1H), 6.63 (s, 1H), 6.00-5.97 (m, 2H), 5.72-5.70 (m, 1H), 4.70-4.66 (m, 1H), 4.46-4.35 (m, 2H), 4.24-4.20 (m, 3H), 3.55-3.45 (m, 2H), 3.26 (s, 1H), 3.24 (s, 1H), 2.94-2.80 (m, 2H), 2.06 (s, 3H).	(CD₃OD) δ 170.1, 149.4, 147.3, 138.7, 138.4, 136.1, 134.4, 134.2, 128.8, 127.6, 123.1, 122.1, 109.1, 103.9, 101.9, 97.1, 71.0, 70.3, 66.8, 66.7, 62.7, 60.7, 36.8, 29.4, 19.3.	95.1%	[M]⁺: C₂₅H₂₅O₅NI 546.077 2 546.074 7
9b	24%	208-210	36.00 (0.025)	(CD₃OD) δ 7.62 (d, J = 1.8 Hz, 2H), 7.54 (d, J = 1.8 Hz, 1H), 6.53 (s, 1H), 6.52 (s, 1H), 5.95-5.90 (m, 2H), 5.68-5.66 (m, 1H), 4.87 (d, J = 13.9 Hz, 1H), 4.73 (d, J = 13.3 Hz, 1H), 4.61-4.50 (m, 2H), 4.46 (d, J = 11.6 Hz, 1H), 4.23-4.21 (m, 1H), 4.07-3.93 (m, 1H), 3.80-3.73 (m, 1H), 3.60-3.49 (m, 1H), 3.23-3.18 (m, 1H), 3.02-2.95 (m, 1H), 2.93-2.86 (m, 1H), 2.13 (s, 3H).	(CD₃OD) δ 171.5, 150.5, 148.4, 137.7, 137.2, 135.8 (2), 135.7, 133.7, 129.6, 123.6 (2), 109.8, 108.5, 105.0, 103.3, 71.8, 68.7, 68.3, 63.5, 62.1, 60.8, 53.3, 38.2, 30.9.	97.0%	[M]⁺: C₂₅H₂₄O₅NBr₂ 576.001 6 575.999 0 (⁷⁹Br+⁷⁹Br) 577.996 6 (⁷⁹Br+⁸¹Br) 579.994 8 (⁸¹Br+⁸¹Br)

Compd.

Yield /%

mp/℃

$ {\left[\alpha \right]}_{\mathrm{D}}^{25\ ^\circ {\rm{C}}} $

(g·100 mL^-1, MeOH)

¹H NMR (600 MHz)

¹³C NMR (151 MHz)

HPLC purity

HR-ESI-MS (m/z)

Formula Calcd. Found

28%

51-53

-10.00
(0.030)

(CDCl₃) δ 6.86 (s, 1H), 6.59 (s, 1H), 5.96-5.88 (m, 2H), 5.56 (s, 1H), 4.58 (s, 1H), 4.11 (d, J = 14.0 Hz, 1H), 3.92-3.88 (m, 1H), 3.67-3.60 (m, 2H), 3.50 (d, J = 14.0 Hz, 1H), 3.33-3.30 (m, 1H), 2.71 (s, 2H), 2.66-2.57 (m, 2H), 2.36 (q, J = 8.7 Hz, 1H), 1.65-1.52 (m, 2H), 1.45-1.32 (m, 2H), 0.93 (t, J = 7.4 Hz, 3H).

(CDCl₃) δ 146.7, 146.3, 143.5, 130.6, 127.7, 116.0, 107.9, 104.6, 101.0, 79.4, 69.9, 69.0, 61.0, 57.1, 53.9, 41.8, 32.2, 28.8, 19.4, 14.0.

96.9%

[M+H]⁺:
C₂₀H₂₆O₄N
344.185 6
344.184 0

20%

114-116

40.91
(0.022)

(CDCl₃) δ 6.86 (s, 1H), 6.60 (s, 1H), 5.96-5.91 (m, 2H), 5.56 (s, 1H), 4.58 (s, 1H), 4.12 (d, J = 14.1 Hz, 1H), 3.92 (s, 1H), 3.72-3.63 (m, 2H), 3.60-3.47 (m, 4H), 3.35-3.32 (m, 1H), 2.71 (s, 2H), 2.66-2.57 (m, 2H), 1.91-1.84 (m, 2H), 1.78-1.73 (m, 2H).

(CDCl₃) δ 146.7, 146.4, 143.7, 130.6, 127.5, 115.8, 107.9, 104.6, 101.1, 79.4, 70.2, 69.1, 61.0, 57.1, 53.9, 45.0, 41.8, 29.5, 28.8, 27.5.

95.2%

[M+H]⁺:
C₂₀H₂₅O₄NCl 378.146 7
378.145 2

45%

64-66

-5.56
(0.018)

(DMSO-d₆) δ 6.87 (s, 1H), 6.66 (s, 1H), 5.94 (d, J = 5.3 Hz, 2H), 5.85-5.79 (m, 1H), 5.42-5.39 (m, 1H), 5.01 (dd, J = 17.2, 1.9 Hz, 1H), 4.95 (dd, J = 17.2, 1.9 Hz, 1H), 4.92-4.75 (m, 1H), 4.41 (s, 1H), 4.00 (d, J = 14.1 Hz, 1H), 3.73-3.70 (m, 1H), 3.62 (dt, J = 9.4, 6.2 Hz, 1H), 3.52 (dt, J = 9.4, 6.2 Hz, 1H), 3.43-3.25 (m, 2H), 3.21-3.14 (m, 1H), 2.59 (d, J = 10.4 Hz, 1H), 2.47-2.39 (m, 1H), 2.36 (d, J = 10.4 Hz, 1H), 2.20 (q, J = 8.6 Hz, 1H), 2.07 (q, J = 7.4 Hz, 2H), 1.64-1.53 (m, 2H).

(DMSO-d₆) δ 146.2, 145.7, 143.6, 138.9, 130.1, 129.6, 116.3, 115.4, 107.5, 105.6, 101.1, 80.3, 68.8, 66.9, 61.1, 57.0, 53.6, 41.3, 30.3, 29.2, 28.6.

98.8%

[M+H]⁺:
C₂₁H₂₆O₄N
356.185 6
356.184 1

30%

102-104

64.71
(0.017)

(CD₃OD) δ 7.03-6.98 (m, 2H), 5.99 (d, J = 1.2 Hz, 1H), 5.97 (d, J = 1.2 Hz, 1H) 5.74-5.70 (m, 1H), 5.68-5.56 (m, 1H), 5.14 (dt, J = 10.1, 1.4 Hz, 1H), 5.07 (dt, J = 10.1, 1.4 Hz, 1H), 4.77-4.75 (m, 1H), 4.55-4.52 (m, 1H), 4.26-4.15 (m, 3H), 3.92-3.83 (m, 1H), 3.56-3.51 (m, 1H), 3.38-3.33 (m, 1H), 3.08 (d, J = 2.4 Hz, 1H), 3.02-2.97 (m, 2H), 2.85-2.78 (m, 1H), 2.53-2.48 (m, 2H).

(CD₃OD) δ 150.8, 148.5, 137.5, 133.1, 131.6, 124.8, 123.5, 119.6, 109.7, 106.7, 103.1, 73.2, 71.6, 69.9, 64.9, 64.1, 62.6, 39.6, 31.0, 29.1.

98.0%

[M]⁺:
C₂₀H₂₄O₄N
342.170 0
342.170 0

20%

150-152

65.22
(0.023)

(CD₃OD) δ 7.00 (s, 1H), 6.98 (s, 1H), 5.99 (d, J = 1.2 Hz, 1H), 5.97 (d, J = 1.2 Hz, 1H), 5.75-5.67 (m, 2H), 5.04-4.87 (m, 2H), 4.73 (d, J = 13.8 Hz, 1H), 4.53-4.51 (m, 1H), 4.24-4.08 (m, 3H), 3.54-3.47 (m, 1H), 3.29 (s, 1H), 3.21-3.10 (m, 1H), 3.01-2.93 (m, 3H), 2.84-2.78 (m, 1H), 2.05-1.93 (m, 2H), 1.86-1.76 (m, 2H).

(CD₃OD) δ 150.8, 148.4, 137.5, 137.4, 131.6, 124.8, 123.4, 117.0, 109.8, 106.7, 103.1, 73.3, 71.5, 69.9, 65.5, 63.9, 62.4, 39.5, 31.2, 31.0, 23.8.

95.5%

[M]⁺:
C₂₁H₂₆O₄N
356.185 6
356.184 4

25%

250-252

69.57
(0.023)

(CD₃OD) δ 6.74-6.70 (m, 2H), 5.71-5.66 (m, 2H), 5.45 (d, J = 3.5 Hz, 1H), 4.50-4.44 (m, 2H), 4.27-4.22 (m, 1H), 3.97-3.84 (m, 3H), 2.97-2.92 (m, 4H), 2.77-2.67 (m, 3H), 2.58-2.51 (m, 1H), 1.68-1.53 (m, 2H), 1.46-1.34 (m, 2H).

(CD₃OD) δ 150.8, 148.5, 137.5, 131.6, 124.8, 123.4, 109.8, 106.7, 103.1, 73.3, 71.6, 69.9, 65.0, 64.0, 62.4, 44.6, 39.6, 31.0, 30.2, 22.2.

98.0%

[M]⁺:
C₂₀H₂₅O₄NCl 378.146 7
378.144 9

20%

170-172

7.50
(0.040)

(CD₃OD) δ 7.00 (s, 1H), 6.99 (s, 1H), 5.97 (d, J = 3.8 Hz, 2H), 5.74-5.70 (m, 2H), 4.91 (dd, J = 16.5, 2.2 Hz, 1H), 4.85 (dd, J = 10.5, 2.1 Hz, 1H), 4.77-4.72 (m, 3H), 4.52 (s, 2H), 4.23-4.12 (m, 3H), 3.53 (td, J = 12.8, 6.6 Hz, 1H), 3.17 (td, J = 12.4, 4.5 Hz, 1H), 3.01-2.94 (m, 3H), 2.83-2.77 (m, 1H), 1.98-1.95 (m, 2H), 1.75-1.65 (m, 2H), 1.26-1.18 (m, 10H).

(CD₃OD) δ 150.7, 148.4, 140.1, 137.6, 131.6, 124.9, 123.3, 114.7, 109.7, 106.7, 103.1, 73.1, 71.6, 69.9, 66.1, 63.9, 62.4, 39.5, 34.8, 31.0, 30.2, 30.1, 30.0 (2), 27.3, 24.5.

95.2%

[M]⁺:
C₂₆H₃₆O₄N
426.263 9
426.262 3

20%

213-215

66.67
(0.030)

(CD₃OD) δ 7.39-7.34 (m, 2H), 7.30 (d, J = 7.9 Hz, 2H), 7.11-6.95 (m, 2H), 6.00 (dd, J = 6.7, 1.3 Hz, 2H), 5.72-5.63 (m, 1H), 4.65-4.48 (m, 2H), 4.27 (d, J = 13.2 Hz, 1H), 4.22-4.09 (m, 5H), 3.42-3.35 (m, 1H), 2.98 (d, J = 11.4 Hz, 1H), 2.74-2.54 (m, 2H), 2.35 (s, 3H).

(CD₃OD) δ 150.8, 148.5, 142.8, 137.6, 134.1 (2), 131.9, 131.3 (2), 126.4, 124.6, 123.4, 110.2, 106.8, 103.2, 72.6, 71.4, 69.9, 68.3, 62.9, 62.4, 39.5, 30.9, 21.3.

96.2%

[M]⁺:
C₂₄H₂₆O₄N
392.185 6
392.184 6

50%

214-216

29.03
(0.031)

(CD₃OD) δ 7.84-7.80 (m, 2H), 7.25-7.21 (m, 2H), 7.01 (s, 1H), 6.95 (s, 1H), 6.01 (d, J = 1.2 Hz, 1H), 5.99 (d, J = 1.2 Hz, 1H), 5.72-5.69 (m, 1H), 4.66-4.59 (m, 1H), 4.55-4.53 (m, 1H), 4.31 (d, J = 13.3 Hz, 1H), 4.25 (d, J = 13.3 Hz, 1H), 4.20-4.14 (m, 4H), 3.47-3.40 (m, 1H), 2.97 (d, J = 11.7 Hz, 1H), 2.76-2.69 (m, 2H).

(CD₃OD) δ 150.7, 148.4, 139.9 (2), 137.5, 135.8 (2), 131.7, 129.1, 124.5, 123.6, 110.1, 106.8, 103.2, 98.5, 72.8, 71.4, 69.8, 67.9, 63.6, 62.6, 39.4, 30.8.

96.2%

[M]⁺:
C₂₃H₂₃O₄NI 504.066 6
504.065 0

32%

63-65

8.57
(0.035)

(CD₃OD) δ 7.47-7.38 (m, 2H), 7.25-7.11 (m, 2H), 6.81 (s, 1H), 6.56 (s, 1H), 5.89-5.79 (m, 2H), 5.56-5.53 (m, 1H), 4.73-4.64 (m, 2H), 4.59-4.56 (m, 1H), 4.06-4.02 (m, 2H), 3.94-3.92 (m, 1H), 3.49-3.43 (m, 1H), 2.93 (s, 1H), 2.83-2.77 (m, 2H), 2.67-2.57 (m, 2H), 2.56-2.50 (m, 1H), 2.36 (q, J = 8.9 Hz, 1H).

(CD₃OD) δ 148.1, 147.7, 144.7, 139.6, 130.4, 130.3 (2), 129.6, 122.8, 122.0 (2), 121.1, 117.3, 108.2, 106.1, 102.3, 80.9, 71.7, 68.9, 62.4, 57.8, 54.7, 42.2, 29.3.

97.5%

[M]⁺:
C₂₄H₂₃O₅NF₃ 462.152 3
462.150 3

44%

210-212

46.43
(0.028)

(CD₃OD) δ 7.68 (d, J = 1.7 Hz, 1H), 7.62 (d, J = 1.7 Hz, 2H), 6.98 (s, 1H), 6.70 (s, 1H), 6.00–5.94 (m, 2H), 5.75-5.71 (m, 1H), 4.69 (d, J = 14.1 Hz, 1H), 4.53-4.51 (m, 1H), 4.44-4.39 (m, 2H), 4.31-4.18 (m, 4H), 3.66-3.53 (m, 1H), 2.94 (d, J = 11.5 Hz, 1H), 2.82-2.78 (m, 2H).

(CD₃OD) δ 150.6, 148.3, 137.4, 135.8 (3), 135.5, 133.6, 131.5, 124.6, 123.7 (2), 109.6, 106.6, 103.2, 72.4, 71.4, 69.8, 67.7, 65.7, 63.3, 54.2, 30.8.

98.0%

[M]⁺:
C₂₃H₂₂O₄NBr₂ 533.991 0
533.989 3 (⁷⁹Br+⁷⁹Br)
535.987 4 (⁷⁹Br+⁸¹Br)
537.985 2 (⁸¹Br+⁸¹Br)

24%

194-196

17.39
(0.023)

(CD₃OD) δ 7.84-7.81 (m, 2H), 7.26-7.20 (m, 2H), 6.93 (s, 1H), 6.63 (s, 1H), 6.00-5.97 (m, 2H), 5.72-5.70 (m, 1H), 4.70-4.66 (m, 1H), 4.46-4.35 (m, 2H), 4.24-4.20 (m, 3H), 3.55-3.45 (m, 2H), 3.26 (s, 1H), 3.24 (s, 1H), 2.94-2.80 (m, 2H), 2.06 (s, 3H).

(CD₃OD) δ 170.1, 149.4, 147.3, 138.7, 138.4, 136.1, 134.4, 134.2, 128.8, 127.6, 123.1, 122.1, 109.1, 103.9, 101.9, 97.1, 71.0, 70.3, 66.8, 66.7, 62.7, 60.7, 36.8, 29.4, 19.3.

95.1%

[M]⁺:
C₂₅H₂₅O₅NI 546.077 2
546.074 7

24%

208-210

36.00
(0.025)

(CD₃OD) δ 7.62 (d, J = 1.8 Hz, 2H), 7.54 (d, J = 1.8 Hz, 1H), 6.53 (s, 1H), 6.52 (s, 1H), 5.95-5.90 (m, 2H), 5.68-5.66 (m, 1H), 4.87 (d, J = 13.9 Hz, 1H), 4.73 (d, J = 13.3 Hz, 1H), 4.61-4.50 (m, 2H), 4.46 (d, J = 11.6 Hz, 1H), 4.23-4.21 (m, 1H), 4.07-3.93 (m, 1H), 3.80-3.73 (m, 1H), 3.60-3.49 (m, 1H), 3.23-3.18 (m, 1H), 3.02-2.95 (m, 1H), 2.93-2.86 (m, 1H), 2.13 (s, 3H).

(CD₃OD) δ 171.5, 150.5, 148.4, 137.7, 137.2, 135.8 (2), 135.7, 133.7, 129.6, 123.6 (2), 109.8, 108.5, 105.0, 103.3, 71.8, 68.7, 68.3, 63.5, 62.1, 60.8, 53.3, 38.2, 30.9.

97.0%

[M]⁺:
C₂₅H₂₄O₅NBr₂ 576.001 6
575.999 0 (⁷⁹Br+⁷⁹Br)
577.996 6 (⁷⁹Br+⁸¹Br)
579.994 8 (⁸¹Br+⁸¹Br)

Compd.	Yield /%	mp/℃	$ {\left[\alpha \right]}_{\mathrm{D}}^{25\ ^\circ {\rm{C}}} $ (g·100 mL^-1, MeOH)	¹H NMR (600 MHz)	¹³C NMR (151 MHz)	HPLC purity	HR-ESI-MS (m/z)
5a	28%	51-53	-10.00 (0.030)	(CDCl₃) δ 6.86 (s, 1H), 6.59 (s, 1H), 5.96-5.88 (m, 2H), 5.56 (s, 1H), 4.58 (s, 1H), 4.11 (d, J = 14.0 Hz, 1H), 3.92-3.88 (m, 1H), 3.67-3.60 (m, 2H), 3.50 (d, J = 14.0 Hz, 1H), 3.33-3.30 (m, 1H), 2.71 (s, 2H), 2.66-2.57 (m, 2H), 2.36 (q, J = 8.7 Hz, 1H), 1.65-1.52 (m, 2H), 1.45-1.32 (m, 2H), 0.93 (t, J = 7.4 Hz, 3H).	(CDCl₃) δ 146.7, 146.3, 143.5, 130.6, 127.7, 116.0, 107.9, 104.6, 101.0, 79.4, 69.9, 69.0, 61.0, 57.1, 53.9, 41.8, 32.2, 28.8, 19.4, 14.0.	96.9%	[M+H]⁺: C₂₀H₂₆O₄N 344.185 6 344.184 0
5b	20%	114-116	40.91 (0.022)	(CDCl₃) δ 6.86 (s, 1H), 6.60 (s, 1H), 5.96-5.91 (m, 2H), 5.56 (s, 1H), 4.58 (s, 1H), 4.12 (d, J = 14.1 Hz, 1H), 3.92 (s, 1H), 3.72-3.63 (m, 2H), 3.60-3.47 (m, 4H), 3.35-3.32 (m, 1H), 2.71 (s, 2H), 2.66-2.57 (m, 2H), 1.91-1.84 (m, 2H), 1.78-1.73 (m, 2H).	(CDCl₃) δ 146.7, 146.4, 143.7, 130.6, 127.5, 115.8, 107.9, 104.6, 101.1, 79.4, 70.2, 69.1, 61.0, 57.1, 53.9, 45.0, 41.8, 29.5, 28.8, 27.5.	95.2%	[M+H]⁺: C₂₀H₂₅O₄NCl 378.146 7 378.145 2
5c	45%	64-66	-5.56 (0.018)	(DMSO-d₆) δ 6.87 (s, 1H), 6.66 (s, 1H), 5.94 (d, J = 5.3 Hz, 2H), 5.85-5.79 (m, 1H), 5.42-5.39 (m, 1H), 5.01 (dd, J = 17.2, 1.9 Hz, 1H), 4.95 (dd, J = 17.2, 1.9 Hz, 1H), 4.92-4.75 (m, 1H), 4.41 (s, 1H), 4.00 (d, J = 14.1 Hz, 1H), 3.73-3.70 (m, 1H), 3.62 (dt, J = 9.4, 6.2 Hz, 1H), 3.52 (dt, J = 9.4, 6.2 Hz, 1H), 3.43-3.25 (m, 2H), 3.21-3.14 (m, 1H), 2.59 (d, J = 10.4 Hz, 1H), 2.47-2.39 (m, 1H), 2.36 (d, J = 10.4 Hz, 1H), 2.20 (q, J = 8.6 Hz, 1H), 2.07 (q, J = 7.4 Hz, 2H), 1.64-1.53 (m, 2H).	(DMSO-d₆) δ 146.2, 145.7, 143.6, 138.9, 130.1, 129.6, 116.3, 115.4, 107.5, 105.6, 101.1, 80.3, 68.8, 66.9, 61.1, 57.0, 53.6, 41.3, 30.3, 29.2, 28.6.	98.8%	[M+H]⁺: C₂₁H₂₆O₄N 356.185 6 356.184 1
6c	30%	102-104	64.71 (0.017)	(CD₃OD) δ 7.03-6.98 (m, 2H), 5.99 (d, J = 1.2 Hz, 1H), 5.97 (d, J = 1.2 Hz, 1H) 5.74-5.70 (m, 1H), 5.68-5.56 (m, 1H), 5.14 (dt, J = 10.1, 1.4 Hz, 1H), 5.07 (dt, J = 10.1, 1.4 Hz, 1H), 4.77-4.75 (m, 1H), 4.55-4.52 (m, 1H), 4.26-4.15 (m, 3H), 3.92-3.83 (m, 1H), 3.56-3.51 (m, 1H), 3.38-3.33 (m, 1H), 3.08 (d, J = 2.4 Hz, 1H), 3.02-2.97 (m, 2H), 2.85-2.78 (m, 1H), 2.53-2.48 (m, 2H).	(CD₃OD) δ 150.8, 148.5, 137.5, 133.1, 131.6, 124.8, 123.5, 119.6, 109.7, 106.7, 103.1, 73.2, 71.6, 69.9, 64.9, 64.1, 62.6, 39.6, 31.0, 29.1.	98.0%	[M]⁺: C₂₀H₂₄O₄N 342.170 0 342.170 0
6d	20%	150-152	65.22 (0.023)	(CD₃OD) δ 7.00 (s, 1H), 6.98 (s, 1H), 5.99 (d, J = 1.2 Hz, 1H), 5.97 (d, J = 1.2 Hz, 1H), 5.75-5.67 (m, 2H), 5.04-4.87 (m, 2H), 4.73 (d, J = 13.8 Hz, 1H), 4.53-4.51 (m, 1H), 4.24-4.08 (m, 3H), 3.54-3.47 (m, 1H), 3.29 (s, 1H), 3.21-3.10 (m, 1H), 3.01-2.93 (m, 3H), 2.84-2.78 (m, 1H), 2.05-1.93 (m, 2H), 1.86-1.76 (m, 2H).	(CD₃OD) δ 150.8, 148.4, 137.5, 137.4, 131.6, 124.8, 123.4, 117.0, 109.8, 106.7, 103.1, 73.3, 71.5, 69.9, 65.5, 63.9, 62.4, 39.5, 31.2, 31.0, 23.8.	95.5%	[M]⁺: C₂₁H₂₆O₄N 356.185 6 356.184 4
6e	25%	250-252	69.57 (0.023)	(CD₃OD) δ 6.74-6.70 (m, 2H), 5.71-5.66 (m, 2H), 5.45 (d, J = 3.5 Hz, 1H), 4.50-4.44 (m, 2H), 4.27-4.22 (m, 1H), 3.97-3.84 (m, 3H), 2.97-2.92 (m, 4H), 2.77-2.67 (m, 3H), 2.58-2.51 (m, 1H), 1.68-1.53 (m, 2H), 1.46-1.34 (m, 2H).	(CD₃OD) δ 150.8, 148.5, 137.5, 131.6, 124.8, 123.4, 109.8, 106.7, 103.1, 73.3, 71.6, 69.9, 65.0, 64.0, 62.4, 44.6, 39.6, 31.0, 30.2, 22.2.	98.0%	[M]⁺: C₂₀H₂₅O₄NCl 378.146 7 378.144 9
6f	20%	170-172	7.50 (0.040)	(CD₃OD) δ 7.00 (s, 1H), 6.99 (s, 1H), 5.97 (d, J = 3.8 Hz, 2H), 5.74-5.70 (m, 2H), 4.91 (dd, J = 16.5, 2.2 Hz, 1H), 4.85 (dd, J = 10.5, 2.1 Hz, 1H), 4.77-4.72 (m, 3H), 4.52 (s, 2H), 4.23-4.12 (m, 3H), 3.53 (td, J = 12.8, 6.6 Hz, 1H), 3.17 (td, J = 12.4, 4.5 Hz, 1H), 3.01-2.94 (m, 3H), 2.83-2.77 (m, 1H), 1.98-1.95 (m, 2H), 1.75-1.65 (m, 2H), 1.26-1.18 (m, 10H).	(CD₃OD) δ 150.7, 148.4, 140.1, 137.6, 131.6, 124.9, 123.3, 114.7, 109.7, 106.7, 103.1, 73.1, 71.6, 69.9, 66.1, 63.9, 62.4, 39.5, 34.8, 31.0, 30.2, 30.1, 30.0 (2), 27.3, 24.5.	95.2%	[M]⁺: C₂₆H₃₆O₄N 426.263 9 426.262 3
6g	20%	213-215	66.67 (0.030)	(CD₃OD) δ 7.39-7.34 (m, 2H), 7.30 (d, J = 7.9 Hz, 2H), 7.11-6.95 (m, 2H), 6.00 (dd, J = 6.7, 1.3 Hz, 2H), 5.72-5.63 (m, 1H), 4.65-4.48 (m, 2H), 4.27 (d, J = 13.2 Hz, 1H), 4.22-4.09 (m, 5H), 3.42-3.35 (m, 1H), 2.98 (d, J = 11.4 Hz, 1H), 2.74-2.54 (m, 2H), 2.35 (s, 3H).	(CD₃OD) δ 150.8, 148.5, 142.8, 137.6, 134.1 (2), 131.9, 131.3 (2), 126.4, 124.6, 123.4, 110.2, 106.8, 103.2, 72.6, 71.4, 69.9, 68.3, 62.9, 62.4, 39.5, 30.9, 21.3.	96.2%	[M]⁺: C₂₄H₂₆O₄N 392.185 6 392.184 6
6h	50%	214-216	29.03 (0.031)	(CD₃OD) δ 7.84-7.80 (m, 2H), 7.25-7.21 (m, 2H), 7.01 (s, 1H), 6.95 (s, 1H), 6.01 (d, J = 1.2 Hz, 1H), 5.99 (d, J = 1.2 Hz, 1H), 5.72-5.69 (m, 1H), 4.66-4.59 (m, 1H), 4.55-4.53 (m, 1H), 4.31 (d, J = 13.3 Hz, 1H), 4.25 (d, J = 13.3 Hz, 1H), 4.20-4.14 (m, 4H), 3.47-3.40 (m, 1H), 2.97 (d, J = 11.7 Hz, 1H), 2.76-2.69 (m, 2H).	(CD₃OD) δ 150.7, 148.4, 139.9 (2), 137.5, 135.8 (2), 131.7, 129.1, 124.5, 123.6, 110.1, 106.8, 103.2, 98.5, 72.8, 71.4, 69.8, 67.9, 63.6, 62.6, 39.4, 30.8.	96.2%	[M]⁺: C₂₃H₂₃O₄NI 504.066 6 504.065 0
6i	32%	63-65	8.57 (0.035)	(CD₃OD) δ 7.47-7.38 (m, 2H), 7.25-7.11 (m, 2H), 6.81 (s, 1H), 6.56 (s, 1H), 5.89-5.79 (m, 2H), 5.56-5.53 (m, 1H), 4.73-4.64 (m, 2H), 4.59-4.56 (m, 1H), 4.06-4.02 (m, 2H), 3.94-3.92 (m, 1H), 3.49-3.43 (m, 1H), 2.93 (s, 1H), 2.83-2.77 (m, 2H), 2.67-2.57 (m, 2H), 2.56-2.50 (m, 1H), 2.36 (q, J = 8.9 Hz, 1H).	(CD₃OD) δ 148.1, 147.7, 144.7, 139.6, 130.4, 130.3 (2), 129.6, 122.8, 122.0 (2), 121.1, 117.3, 108.2, 106.1, 102.3, 80.9, 71.7, 68.9, 62.4, 57.8, 54.7, 42.2, 29.3.	97.5%	[M]⁺: C₂₄H₂₃O₅NF₃ 462.152 3 462.150 3
6j	44%	210-212	46.43 (0.028)	(CD₃OD) δ 7.68 (d, J = 1.7 Hz, 1H), 7.62 (d, J = 1.7 Hz, 2H), 6.98 (s, 1H), 6.70 (s, 1H), 6.00–5.94 (m, 2H), 5.75-5.71 (m, 1H), 4.69 (d, J = 14.1 Hz, 1H), 4.53-4.51 (m, 1H), 4.44-4.39 (m, 2H), 4.31-4.18 (m, 4H), 3.66-3.53 (m, 1H), 2.94 (d, J = 11.5 Hz, 1H), 2.82-2.78 (m, 2H).	(CD₃OD) δ 150.6, 148.3, 137.4, 135.8 (3), 135.5, 133.6, 131.5, 124.6, 123.7 (2), 109.6, 106.6, 103.2, 72.4, 71.4, 69.8, 67.7, 65.7, 63.3, 54.2, 30.8.	98.0%	[M]⁺: C₂₃H₂₂O₄NBr₂ 533.991 0 533.989 3 (⁷⁹Br+⁷⁹Br) 535.987 4 (⁷⁹Br+⁸¹Br) 537.985 2 (⁸¹Br+⁸¹Br)
9a	24%	194-196	17.39 (0.023)	(CD₃OD) δ 7.84-7.81 (m, 2H), 7.26-7.20 (m, 2H), 6.93 (s, 1H), 6.63 (s, 1H), 6.00-5.97 (m, 2H), 5.72-5.70 (m, 1H), 4.70-4.66 (m, 1H), 4.46-4.35 (m, 2H), 4.24-4.20 (m, 3H), 3.55-3.45 (m, 2H), 3.26 (s, 1H), 3.24 (s, 1H), 2.94-2.80 (m, 2H), 2.06 (s, 3H).	(CD₃OD) δ 170.1, 149.4, 147.3, 138.7, 138.4, 136.1, 134.4, 134.2, 128.8, 127.6, 123.1, 122.1, 109.1, 103.9, 101.9, 97.1, 71.0, 70.3, 66.8, 66.7, 62.7, 60.7, 36.8, 29.4, 19.3.	95.1%	[M]⁺: C₂₅H₂₅O₅NI 546.077 2 546.074 7
9b	24%	208-210	36.00 (0.025)	(CD₃OD) δ 7.62 (d, J = 1.8 Hz, 2H), 7.54 (d, J = 1.8 Hz, 1H), 6.53 (s, 1H), 6.52 (s, 1H), 5.95-5.90 (m, 2H), 5.68-5.66 (m, 1H), 4.87 (d, J = 13.9 Hz, 1H), 4.73 (d, J = 13.3 Hz, 1H), 4.61-4.50 (m, 2H), 4.46 (d, J = 11.6 Hz, 1H), 4.23-4.21 (m, 1H), 4.07-3.93 (m, 1H), 3.80-3.73 (m, 1H), 3.60-3.49 (m, 1H), 3.23-3.18 (m, 1H), 3.02-2.95 (m, 1H), 2.93-2.86 (m, 1H), 2.13 (s, 3H).	(CD₃OD) δ 171.5, 150.5, 148.4, 137.7, 137.2, 135.8 (2), 135.7, 133.7, 129.6, 123.6 (2), 109.8, 108.5, 105.0, 103.3, 71.8, 68.7, 68.3, 63.5, 62.1, 60.8, 53.3, 38.2, 30.9.	97.0%	[M]⁺: C₂₅H₂₄O₅NBr₂ 576.001 6 575.999 0 (⁷⁹Br+⁷⁹Br) 577.996 6 (⁷⁹Br+⁸¹Br) 579.994 8 (⁸¹Br+⁸¹Br)

Compd.

Yield /%

mp/℃

$ {\left[\alpha \right]}_{\mathrm{D}}^{25\ ^\circ {\rm{C}}} $

(g·100 mL^-1, MeOH)

¹H NMR (600 MHz)

¹³C NMR (151 MHz)

HPLC purity

HR-ESI-MS (m/z)

Formula Calcd. Found

28%

51-53

-10.00
(0.030)

(CDCl₃) δ 146.7, 146.3, 143.5, 130.6, 127.7, 116.0, 107.9, 104.6, 101.0, 79.4, 69.9, 69.0, 61.0, 57.1, 53.9, 41.8, 32.2, 28.8, 19.4, 14.0.

96.9%

[M+H]⁺:
C₂₀H₂₆O₄N
344.185 6
344.184 0

20%

114-116

40.91
(0.022)

(CDCl₃) δ 146.7, 146.4, 143.7, 130.6, 127.5, 115.8, 107.9, 104.6, 101.1, 79.4, 70.2, 69.1, 61.0, 57.1, 53.9, 45.0, 41.8, 29.5, 28.8, 27.5.

95.2%

[M+H]⁺:
C₂₀H₂₅O₄NCl 378.146 7
378.145 2

45%

64-66

-5.56
(0.018)

(DMSO-d₆) δ 146.2, 145.7, 143.6, 138.9, 130.1, 129.6, 116.3, 115.4, 107.5, 105.6, 101.1, 80.3, 68.8, 66.9, 61.1, 57.0, 53.6, 41.3, 30.3, 29.2, 28.6.

98.8%

[M+H]⁺:
C₂₁H₂₆O₄N
356.185 6
356.184 1

30%

102-104

64.71
(0.017)

(CD₃OD) δ 150.8, 148.5, 137.5, 133.1, 131.6, 124.8, 123.5, 119.6, 109.7, 106.7, 103.1, 73.2, 71.6, 69.9, 64.9, 64.1, 62.6, 39.6, 31.0, 29.1.

98.0%

[M]⁺:
C₂₀H₂₄O₄N
342.170 0
342.170 0

20%

150-152

65.22
(0.023)

(CD₃OD) δ 150.8, 148.4, 137.5, 137.4, 131.6, 124.8, 123.4, 117.0, 109.8, 106.7, 103.1, 73.3, 71.5, 69.9, 65.5, 63.9, 62.4, 39.5, 31.2, 31.0, 23.8.

95.5%

[M]⁺:
C₂₁H₂₆O₄N
356.185 6
356.184 4

25%

250-252

69.57
(0.023)

(CD₃OD) δ 150.8, 148.5, 137.5, 131.6, 124.8, 123.4, 109.8, 106.7, 103.1, 73.3, 71.6, 69.9, 65.0, 64.0, 62.4, 44.6, 39.6, 31.0, 30.2, 22.2.

98.0%

[M]⁺:
C₂₀H₂₅O₄NCl 378.146 7
378.144 9

20%

170-172

7.50
(0.040)

(CD₃OD) δ 150.7, 148.4, 140.1, 137.6, 131.6, 124.9, 123.3, 114.7, 109.7, 106.7, 103.1, 73.1, 71.6, 69.9, 66.1, 63.9, 62.4, 39.5, 34.8, 31.0, 30.2, 30.1, 30.0 (2), 27.3, 24.5.

95.2%

[M]⁺:
C₂₆H₃₆O₄N
426.263 9
426.262 3

20%

213-215

66.67
(0.030)

(CD₃OD) δ 150.8, 148.5, 142.8, 137.6, 134.1 (2), 131.9, 131.3 (2), 126.4, 124.6, 123.4, 110.2, 106.8, 103.2, 72.6, 71.4, 69.9, 68.3, 62.9, 62.4, 39.5, 30.9, 21.3.

96.2%

[M]⁺:
C₂₄H₂₆O₄N
392.185 6
392.184 6

50%

214-216

29.03
(0.031)

(CD₃OD) δ 150.7, 148.4, 139.9 (2), 137.5, 135.8 (2), 131.7, 129.1, 124.5, 123.6, 110.1, 106.8, 103.2, 98.5, 72.8, 71.4, 69.8, 67.9, 63.6, 62.6, 39.4, 30.8.

96.2%

[M]⁺:
C₂₃H₂₃O₄NI 504.066 6
504.065 0

32%

63-65

8.57
(0.035)

(CD₃OD) δ 148.1, 147.7, 144.7, 139.6, 130.4, 130.3 (2), 129.6, 122.8, 122.0 (2), 121.1, 117.3, 108.2, 106.1, 102.3, 80.9, 71.7, 68.9, 62.4, 57.8, 54.7, 42.2, 29.3.

97.5%

[M]⁺:
C₂₄H₂₃O₅NF₃ 462.152 3
462.150 3

44%

210-212

46.43
(0.028)

(CD₃OD) δ 150.6, 148.3, 137.4, 135.8 (3), 135.5, 133.6, 131.5, 124.6, 123.7 (2), 109.6, 106.6, 103.2, 72.4, 71.4, 69.8, 67.7, 65.7, 63.3, 54.2, 30.8.

98.0%

[M]⁺:
C₂₃H₂₂O₄NBr₂ 533.991 0
533.989 3 (⁷⁹Br+⁷⁹Br)
535.987 4 (⁷⁹Br+⁸¹Br)
537.985 2 (⁸¹Br+⁸¹Br)

24%

194-196

17.39
(0.023)

(CD₃OD) δ 170.1, 149.4, 147.3, 138.7, 138.4, 136.1, 134.4, 134.2, 128.8, 127.6, 123.1, 122.1, 109.1, 103.9, 101.9, 97.1, 71.0, 70.3, 66.8, 66.7, 62.7, 60.7, 36.8, 29.4, 19.3.

95.1%

[M]⁺:
C₂₅H₂₅O₅NI 546.077 2
546.074 7

24%

208-210

36.00
(0.025)

(CD₃OD) δ 171.5, 150.5, 148.4, 137.7, 137.2, 135.8 (2), 135.7, 133.7, 129.6, 123.6 (2), 109.8, 108.5, 105.0, 103.3, 71.8, 68.7, 68.3, 63.5, 62.1, 60.8, 53.3, 38.2, 30.9.

97.0%

[M]⁺:
C₂₅H₂₄O₅NBr₂ 576.001 6
575.999 0 (⁷⁹Br+⁷⁹Br)
577.996 6 (⁷⁹Br+⁸¹Br)
579.994 8 (⁸¹Br+⁸¹Br)


3	/	/	/	＞50.00	＞200.00	N/A
4	/	/	/	21.81 ± 2.72	154.66 ± 4.15	7.09
5a	H		/	＞50.00	＞200.00	N/A
5b	H		/	＞50.00	83.43 ± 2.80	＜1.67
5c	H		/	＞50.00	＞200.00	N/A
6a	H	H		2.36 ± 0.24	38.98 ± 1.04	16.52
6b	H	H		35.43 ± 9.92	＞200.00	＞5.64
6c	H	H		＞50.00	＞200.00	N/A
6d	H	H		＞50.00	＞200.00	N/A
6e	H	H		33.79 ± 6.55	＞200.00	＞5.92
6f	H	H		3.90 ± 3.00	＞200.00	＞51.28
6g	H	H		＞50.00	65.83 ± 1.94	＜1.32
6h	H	H		23.10 ± 0.40	＞200.00	＞8.66
6i	H	H		6.00 ± 0.80	21.50 ± 1.60	3.58
6j	H	H		5.59 ± 0.63	＞200.00	＞35.78
7	/	/	/	＞50.00	＞200.00	N/A
9a		H		36.50 ± 0.10	＞200.00	＞5.48
9b		H		33.90 ± 1.90	＞200.00	＞5.90
1	/	/	/	0.69 ± 0.06	6.74 ± 0.34	9.77
MPV	/	/	/	0.21 ± 0.01	78.58 ± 26.24	374.19

Compd.

R₁

R₂

R₃

EC₅₀/μmol·L^{-1 a}

CC₅₀/μmol·L^{-1 b}

SI^c

＞50.00

＞200.00

N/A

21.81 ± 2.72

154.66 ± 4.15

7.09

＞50.00

＞200.00

N/A

＞50.00

83.43 ± 2.80

＜1.67

＞50.00

＞200.00

N/A

2.36 ± 0.24

38.98 ± 1.04

16.52

35.43 ± 9.92

＞200.00

＞5.64

＞50.00

＞200.00

N/A

＞50.00

＞200.00

N/A

33.79 ± 6.55

＞200.00

＞5.92

3.90 ± 3.00

＞200.00

＞51.28

＞50.00

65.83 ± 1.94

＜1.32

23.10 ± 0.40

＞200.00

＞8.66

6.00 ± 0.80

21.50 ± 1.60

3.58

5.59 ± 0.63

＞200.00

＞35.78

＞50.00

＞200.00

N/A

36.50 ± 0.10

＞200.00

＞5.48

33.90 ± 1.90

＞200.00

＞5.90

0.69 ± 0.06

6.74 ± 0.34

9.77

MPV

0.21 ± 0.01

78.58 ± 26.24

374.19


3	/	/	/	＞50.00	＞200.00	N/A
4	/	/	/	21.81 ± 2.72	154.66 ± 4.15	7.09
5a	H		/	＞50.00	＞200.00	N/A
5b	H		/	＞50.00	83.43 ± 2.80	＜1.67
5c	H		/	＞50.00	＞200.00	N/A
6a	H	H		2.36 ± 0.24	38.98 ± 1.04	16.52
6b	H	H		35.43 ± 9.92	＞200.00	＞5.64
6c	H	H		＞50.00	＞200.00	N/A
6d	H	H		＞50.00	＞200.00	N/A
6e	H	H		33.79 ± 6.55	＞200.00	＞5.92
6f	H	H		3.90 ± 3.00	＞200.00	＞51.28
6g	H	H		＞50.00	65.83 ± 1.94	＜1.32
6h	H	H		23.10 ± 0.40	＞200.00	＞8.66
6i	H	H		6.00 ± 0.80	21.50 ± 1.60	3.58
6j	H	H		5.59 ± 0.63	＞200.00	＞35.78
7	/	/	/	＞50.00	＞200.00	N/A
9a		H		36.50 ± 0.10	＞200.00	＞5.48
9b		H		33.90 ± 1.90	＞200.00	＞5.90
1	/	/	/	0.69 ± 0.06	6.74 ± 0.34	9.77
MPV	/	/	/	0.21 ± 0.01	78.58 ± 26.24	374.19

Compd.

R₁

R₂

R₃

EC₅₀/μmol·L^{-1 a}

CC₅₀/μmol·L^{-1 b}

SI^c

＞50.00

＞200.00

N/A

21.81 ± 2.72

154.66 ± 4.15

7.09

＞50.00

＞200.00

N/A

＞50.00

83.43 ± 2.80

＜1.67

＞50.00

＞200.00

N/A

2.36 ± 0.24

38.98 ± 1.04

16.52

35.43 ± 9.92

＞200.00

＞5.64

＞50.00

＞200.00

N/A

＞50.00

＞200.00

N/A

33.79 ± 6.55

＞200.00

＞5.92

3.90 ± 3.00

＞200.00

＞51.28

＞50.00

65.83 ± 1.94

＜1.32

23.10 ± 0.40

＞200.00

＞8.66

6.00 ± 0.80

21.50 ± 1.60

3.58

5.59 ± 0.63

＞200.00

＞35.78

＞50.00

＞200.00

N/A

36.50 ± 0.10

＞200.00

＞5.48

33.90 ± 1.90

＞200.00

＞5.90

0.69 ± 0.06

6.74 ± 0.34

9.77

MPV

0.21 ± 0.01

78.58 ± 26.24

374.19

Item	AutoDock		Discovery Studio		MVD
6a-NiRAN catalytic center	-7.33		-27.65		-98.39
6a-NSP12 catalytic center	-6.02		-21.57		-97.94

Item

AutoDock

Discovery Studio

MVD

Binding energy /kcal·mol^-1

CDocker interaction energy/kcal·mol^-1

MolDock score

6a-NiRAN catalytic center

-7.33

-27.65

-98.39

6a-NSP12 catalytic center

-6.02

-21.57

-97.94

Item	AutoDock		Discovery Studio		MVD
6a-NiRAN catalytic center	-7.33		-27.65		-98.39
6a-NSP12 catalytic center	-6.02		-21.57		-97.94

Item

AutoDock

Discovery Studio

MVD

Binding energy /kcal·mol^-1

CDocker interaction energy/kcal·mol^-1

MolDock score

6a-NiRAN catalytic center

-7.33

-27.65

-98.39

6a-NSP12 catalytic center

-6.02

-21.57

-97.94

石蒜碱类衍生物的合成与抗SARS-CoV-2的活性和作用机制研究

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梅钰恒 , 李佳玉 , 宋丹青 , 彭宗根 , 李迎红 ^*

药学学报 | 研究论文 2024,59(2): 395-403

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药学学报 | 研究论文 2024, 59(2): 395-403

石蒜碱类衍生物的合成与抗SARS-CoV-2的活性和作用机制研究

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梅钰恒, 李佳玉, 宋丹青, 彭宗根, 李迎红^*

作者信息

中国医学科学院、北京协和医学院医药生物技术研究所, 北京 100050

通讯作者:

^*李迎红, Tel: 86-10-63165284, E-mail: liyinghong@imb.pumc.edu.cn

Synthesis and anti-SARS-CoV-2 activity and mechanism research of lycorine derivatives

Yu-heng MEI, Jia-yu LI, Dan-qing SONG, Zong-gen PENG, Ying-hong LI^*

Affiliations

Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China

出版时间: 2024-02-12 doi: 10.16438/j.0513-4870.2023-1121

文章导航

摘要

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本研究设计合成了5种不同结构类型共18个石蒜碱衍生物, 在感染HCoV-OC43的H460细胞模型上考察了其抗病毒活性。构效关系结果表明在石蒜碱的6N原子上引入适当的取代基有利于活性的提高。其中, 化合物6a表现出较好的活性, 半数有效浓度(EC₅₀) 和选择指数(SI) 值分别为2.36 μmol·L^-1和16.52。表面等离子共振实验显示6a可能靶向SARS-CoV-2的RNA依赖的RNA聚合酶(RdRp) 的非结构蛋白12 (NSP12) 亚基, 解离常数(K_D) 值为1.36 μmol·L^-1。分子对接结果提示6a可能作用于NSP12的类囊病毒RdRp相关核苷酸转移酶(NiRAN) 催化中心, 与瑞德西韦等拟核苷类药物的作用机制不同。该研究为将石蒜碱类衍生物发展成为一类全新的抗SARS-CoV-2小分子抑制剂提供了科学数据。

关键词

石蒜碱类衍生物 / 新冠病毒 / RNA依赖的RNA聚合酶 / 类囊病毒RdRp相关核苷酸转移酶结构域 / 抗病毒

Abstract

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We designed and synthesized eighteen lycorine derivatives with five different structural types, and evaluated their antiviral activities on a HCoV-OC43-infected H460 cell model. Structure-activity relationships suggested that the introduction of appropriate substituents on the 6N atom of lycorine was beneficial to activity. Compound 6a gave a good activity with the half effective concentration (EC₅₀) and selectivity index (SI) values of 2.36 μmol·L^-1 and 16.52, respectively. Surface plasmon resonance (SPR) result indicated that 6a might target the non-structural protein 12 (NSP12) subunit in RNA-dependent RNA polymerase (RdRp) of SARS-CoV-2 with the dissociation constant (K_D) value of 1.36 μmol·L^-1. Molecular docking indicated that 6a might act on nidovirus RdRp-associated nucleotidyltransferase (NiRAN) catalytic center of NSP12, distinct from the mechanism of nucleoside-like drugs such as remdesivir. This study provides scientific data for the development of lycorine derivatives into a new class of anti-SARS-CoV-2 small molecule inhibitors.

Key words

lycorine derivative / SARS-CoV-2 / RNA-dependent RNA polymerase (RdRp) / nidovirus RdRp-associated nucleotidyltransferase (NiRAN) / antivirus

引用本文

梅钰恒, 李佳玉, 宋丹青, 彭宗根, 李迎红. 石蒜碱类衍生物的合成与抗SARS-CoV-2的活性和作用机制研究. 药学学报, 2024 , 59 (2) : 395 -403 . DOI: 10.16438/j.0513-4870.2023-1121

Yu-heng MEI, Jia-yu LI, Dan-qing SONG, Zong-gen PENG, Ying-hong LI. Synthesis and anti-SARS-CoV-2 activity and mechanism research of lycorine derivatives[J]. Acta Pharmaceutica Sinica, 2024 , 59 (2) : 395 -403 . DOI: 10.16438/j.0513-4870.2023-1121

正文

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新型冠状病毒(SARS-CoV-2) 是一种β属冠状病毒, 自2019年被首次发现以来迅速席卷全球, 对全球卫生体系造成了严重冲击, 给人类社会带来了巨大恐慌和经济负担。截至目前, 由SARS-CoV-2引起的新冠病毒肺炎(COVID-19) 已在全球范围内造成近700万例的死亡^[1]。新冠病毒自发现以来持续性变异, 尤其是最近出现的新型奥密克戎变异株的传染性和免疫逃逸能力不断增强, 由先前感染或接种疫苗建立起的人群免疫屏障变得越发脆弱, 临床治疗COVID-19手段的有效性也受到了限制^[2-6]。RNA依赖的RNA聚合酶(RdRp) 抑制剂瑞德西韦是最早应用于临床的抗COVID-19药物。但是, 随着瑞德西韦在临床的广泛应用, 病毒RdRp的催化中心及周围已经进化出D484Y和E802D等多种突变, 导致了瑞德西韦的耐药^[7-11]。因此, 研发全新的RdRp小分子抑制剂具有重要的意义, 尤其是用于应对新型变异株以及未来随时可能出现的冠状病毒突发事件。

RdRp是由核心蛋白NSP12以及辅助因子NSP7和NSP8组成的蛋白复合体(图 1A), 负责RNA病毒基因组的复制和转录, 在病毒在宿主细胞中的自我复制过程中发挥着非常重要的作用。NSP12负责新生RNA链的延伸, NSP7和NSP8发挥引物酶和稳定NSP12构象的作用^[12]。NSP12蛋白结构中最重要的功能区域是催化中心(图 1A), 即瑞德西韦、莫努匹韦和VV116等拟核苷类抑制剂药物的主要结合位点。瑞德西韦通过共价方式嵌入至引物RNA链中, 引起病毒RNA链延伸过程的错配, 继而终止其延伸^[13]。此外, 类囊病毒RdRp相关核苷酸转移酶(NiRAN) (图 1A) 催化中心也是NSP12蛋白的主要组成部分, 主要负责鸟苷酸转移以及核苷单磷酸向病毒辅因子蛋白NSP7、NSP8和NSP9的转运, 是鸟苷类似物AT-527的最主要作用位点^[14]。以NiRAN结构域为靶点可能成为开发新型抗SARS-CoV-2药物的一个新方向^[14]。

石蒜碱(1, 图 1B) 是从石蒜科植物分离出的一种吡咯并菲啶类生物碱, 具有广泛的抗病毒活性, 对SARS-CoV、SARS-CoV-2、单纯疱疹、脊髓灰质炎、寨卡、丙肝、登革热、人71型肠道、禽流感H5N1等多种病毒均显示出较好的活性^{[15, 16]}。但是, 其抗SARS-CoV-2的作用机制尚不明确, 构效关系研究尚未见文献报道。

因此, 本研究以石蒜碱为先导化合物, 分别对其A环、2位羟基、3位氢键、6位叔胺及7位碳进行结构改造, 共设计合成18个石蒜碱衍生物。以感染同属于β属冠状病毒HCoV-OC43的H460细胞为模型评价了其抗SARS-CoV-2的活性, 系统总结了其抗SARS-CoV-2构效关系, 并对重点化合物开展了初步机制学研究。

结果与讨论

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1 化合物的合成

目标化合物的合成如合成路线1所示。以氨水处理的游离石蒜碱(1) 为原料, 按文献报道的方法合成目标化合物3^[17]、4^[18]、6a^[18]、6b^[18]和7^[19]。

在强碱(NaH) 的作用下, 卤代脂肪烃可选择性地与化合物1的2-OH发生亲核取代生成5a~5c, 这是因为2-OH被强碱拔掉氢原子后会形成氧负离子, 进而通过离域作用与相邻的碳碳双键形成稳定的过渡态, 表现出相较于1-OH更高的反应性。在中性条件下, 化合物1的6N原子更容易被烷基化或苄基化生成相应的6S构型的6N取代石蒜碱季铵盐化合物6c~6j。本研究采用高斯计算的方法, 在B3LYP/6-311G(d, p) 水平, 基于时间依赖性密度泛函理论(TDDFT) 在0.35 eV的半频带宽度下得到代表性化合物6j的理论圆二色(ECD) 光谱, 与实验ECD光谱高度相似, 从而确证了该系列化合物N原子的绝对构型。

化合物1在乙酸酐和吡啶中发生酰化反应生成石蒜碱双酯2。在酸水解的条件下, 由于2位碳原子毗邻的O原子与碳碳双键之间的离域作用, 中间体2会选择性脱去2-乙酰基, 生成中间体8^[20]。中间体8在乙腈中与卤代烃发生亲核取代反应, 可以较高的产率生成目标化合物1-乙酰-6N取代石蒜碱季铵盐9a和9b。所有全新目标化合物结构和纯度经¹H NMR、¹³C NMR、HR-MS及HPLC分析确证。目标化合物的收率、熔点和比旋光度等理化参数及波谱数据见表 1。

2 石蒜碱类衍生物抗HCoV-OC43活性构效关系

因为HCoV-OC43与SARS-CoV-2同属β属冠状病毒, 因此以HCoV-OC43感染的H460细胞为模型, 以molnupiravir EIDD-1931 (MPV) 为阳性对照^[21], 通过Cell Counting Kit-8 (CCK-8) 法考察化合物的抗病毒作用。计算了化合物的半数有效浓度(EC₅₀) 以及在未感染病毒的情况下化合物对细胞本身的半数毒性浓度(CC₅₀), 并以EC₅₀值和SI (CC₅₀/EC₅₀) 值作为判断标准。化合物的EC₅₀值越低, SI值越高, 则化合物的抗病毒活性越强。目标化合物的结构与活性结果见表 2。

考察了先导物1的吡咯并菲啶母核对活性的影响。结果显示, A环开环的石蒜碱衍生物3、3位碳碳双键还原的二氢石蒜碱4以及7位选择性氧化的石蒜碱酰胺衍生物7的活性均大幅降低甚至完全丧失, 提示石蒜碱的吡咯并菲啶母核对于保持其抗病毒活性是必需的。

为了提升石蒜碱类化合物的抗病毒活性, 采用了提高脂溶性的策略。在保留吡咯并菲啶母核的基础上, 在2位羟基上引入不同长度的烷基链, 制备了化合物5a~5c。抗病毒活性结果显示, 相较先导物1, 上述化合物的抗病毒活性均明显降低甚至消失, 提示2位羟基引入烷基可能不利于活性。

尝试从改善水溶性的角度提高化合物的抗病毒活性。在保留吡咯并菲啶母核的基础上, 在6位氮原子上引入不同的脂肪链或取代苄基形成水溶性改善的季铵盐类化合物, 得到6N-取代石蒜碱季铵盐6a~6j。其中, 化合物6a (乙基取代) 和6f (癸烯基取代) 的活性最佳, EC₅₀分别为2.36和3.9 μmol·L^-1, SI值分别为16.52和大于51.28, 与先导物的活性相当, 提示在石蒜碱的6N原子上引入合适的取代基有利于活性的提高。在此基础上, 进一步在1-OH上引入酯基以提高其脂溶性, 得到两个目标化合物9a和9b, 其抗病毒活性均下降。鉴于化合物6a的EC₅₀值最低, 将其作为代表性化合物开展下一步研究。

3 化合物6a与NSP12具有较强的亲和力

石蒜碱通过靶向寨卡病毒的RdRp发挥抗病毒作用^[22], 推测石蒜碱化合物也可能通过靶向SARS-CoV-2的RdRp抑制病毒的增殖。为此, 利用表面等离子共振(SPR) 实验, 考察了6a与SARS-CoV-2 NSP12蛋白之间的直接相互作用, 并测定其亲和力(K_D值)。如图 2所示, 化合物6a能够浓度依赖性地与NSP12结合, K_D值为1.36 μmol·L^-1, 提示二者间存在较强的亲和力。

4 化合物6a可能靶向NSP12的NiRAN结构域催化中心

为了预测6a与NSP12 (PDB ID: 7ed5) 之间的结合模式, 采用AutoDock Tools 1.5.6, Discovery Studio 4.5和Molegro Virtual Docker (MVD) 6.0三个软件, 将6a与NSP12的两个活性腔, 即NiRAN结构域催化中心和NSP12催化中心, 分别进行虚拟对接。三种软件的对接结果趋势一致, 6a与NiRAN催化中心的结合能量值较低, 提示6a与NiRAN催化中心的结合能力优于NSP12催化中心(表 3)。

进一步采用PyMOL对虚拟对接结果进行可视化分析(图 3)。CDocker对接结果显示, 化合物6a与NiRAN催化中心的LYS-50、ASP-208和TYR-217之间均能形成较强的氢键作用(图 3B), 此外, 6a与CYS-53之间的π硫共轭相互作用以及与ASN-209和ASP-218等多个氨基酸残基间的范德华力也可能促进6a与NiRAN结构域之间的稳定结合。相较之下, 化合物6a仅能与NSP12催化中心的ASP-865和GLN-815两个氨基酸形成氢键作用(图 3C)。类似地, AutoDock和MVD的对接结果分析也显示, 相较于NSP12催化中心, 6a与NiRAN催化中心能够形成更多的氢键相互作用(图 3E~H)。

采用Discovery Studio 4.5软件, 将非活性化合物6g与NiRAN催化中心也进行了虚拟对接。CDocker结果显示, 化合物6a能够与ASN-52、HIS-75、LYS-50、ASP-208、ASP-218等氨基酸残基组成的疏水口袋较好地适配(图 3B), 而6g无法进入该口袋, 可能是由于6N上的苄基取代基体积过大的缘故(图 3D), 提示在石蒜碱6N原子上引入合适体积的基团有利于与NiRAN的催化中心的结合, 与本论文前述的构效关系结果相一致。

上述结果表明, 不同于瑞德西韦和莫努匹韦等拟核苷类抑制剂, 化合物6a极有可能通过靶向SARS-CoV-2的NiRAN结构域催化中心发挥抗病毒作用, 未来可能与瑞德西韦等拟核苷类抑制剂联用发挥协同抗病毒作用, 值得进一步研究。

小结

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本文设计合成了18个石蒜碱类衍生物, 其中13个为全新的石蒜碱衍生物, 并初步总结了其抗SARS-CoV-2的构效关系。结果提示: 石蒜碱的吡咯并菲啶母核骨架为活性必需, 在6N原子上引入合适的基团对活性有利。其中, 化合物6a表现出了较好的活性和较低的细胞毒性。进一步研究显示, 6a与SARS-CoV-2 RdRp的NSP12亚基展现出良好的结合亲和力, 且极有可能通过靶向NSP12 NiRAN结构域的催化中心抑制病毒的增殖。鉴于化合物6a的作用靶点与瑞德西韦不同, 因而极有可能与瑞德西韦联用发挥协同抗病毒作用, 从而为开发全新机制的SARS-CoV-2小分子抑制剂提供新的思路。

实验部分

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熔点用MP90熔点仪测定(Mettler toledo, Columbus, USA), 未经校正; 比旋光度用Rudolph Autopol IV-T旋光仪测定(Rudolph Research Analytical, NJ, USA); ¹H NMR和¹³C NMR用Bruker Avance Ⅲ 600 MHz核磁共振仪(Varian, San Francisco, USA) 测定, 溶剂为CDCl₃、DMSO-d₆和CD₃OD (带TMS内标); ESI HR-MS用Autospec Ultima-TOF质谱测定仪(Micromass UK Ltd., Manchester, UK) 测定; 圆二色光谱用Chirascan Plus圆二色谱仪(Applied Photophysics, Surrey, UK) 测定; Flash柱分离纯化用Combiflash Rf 200快速制备液相(Teledyne, Nebraska, USA); 紫外检测用ZF-7A手提紫外检测灯(上海宝山顾村电光仪器厂); 薄层色谱(TLC) 采用Merck 60 F254薄层色谱硅胶板(Merck & Co Inc., Darmstadt, German); 纯度用安捷伦1200高效液相色谱仪(Agilent Technologies, Inc., Santa Clara, USA) 测定。采用Agilent Eclipse Plus C18色谱柱(4.6 mm × 150 mm, 5 μm); 以0.1%磷酸(A)-甲醇(B) 为流动相。5a~5c用55% B等度洗脱10 min; 6c~6j, 9a和9b梯度洗脱(0~7 min, 50%~80% B; 7~11 min, 80% B; 11~13 min, 50% B); 流速为1.0 mL·min^-1; 检测波长289 nm; 柱温30 ℃; 进样量20 μL。

所用试剂均购自泰坦、毕得和通广等试剂公司的分析纯试剂, 未经纯化直接使用。

1 化学合成

1.1 目标化合物5a~5c的合成

向石蒜碱盐酸盐(4 mmol) 的50 mL蒸馏水溶液中加入15 mL饱和氨水, 室温搅拌4 h, 置于4 ℃冷却4 h。抽滤, 用15 mL蒸馏水润洗滤渣3次, 真空干燥, 得到化合物1直接用于下一步反应。向1 (1 mmol) 的10 mL DMF溶液中缓慢加入NaH (2 mmol), 室温搅拌20 min, 缓慢滴加溴代脂肪烃, 室温搅拌20 h。TLC监测反应完成后, 加入冰水(20 mL) 淬灭反应, 加入二氯甲烷(30 mL) 萃取3次, 合并有机相, 分别用饱和NH₄Cl溶液(100 mL), 饱和NaCl溶液(100 mL) 洗涤, 无水Na₂SO₄干燥, 过滤, 减压浓缩后硅胶匀化, 以石油醚和乙酸乙酯为流动相, 经Flash快速柱色谱分离, 得到目标化合物5a~5c。

1.2 目标化合物6c~6j的合成

向1 (0.5 mmol) 的5 mL DMF溶液中加入卤代烃(0.7 mmol), 60 ℃搅拌10 h, TLC监测反应完成后, 减压浓缩。残余物用蒸馏水(50 mL) 溶解, 用二氯甲烷(50 mL) 洗涤两次, 浓缩, 得到目标化合物6c~6j。

1.3 目标化合物9a和9b的合成

向2^[20] (0.5 mmol) 的15 mL甲醇溶液中加入浓盐酸(6 mL), 55 ℃搅拌1 h。用饱和NaHCO₃溶液将混合体系的pH值调至8, 加入二氯甲烷(20 mL), 用饱和NaCl溶液(20 mL) 洗涤, 用无水Na₂SO₄干燥, 过滤, 减压浓缩后硅胶匀化, 以乙酸乙酯和甲醇为流动相, 经Flash快速柱色谱分离, 得到化合物8。

向8 (0.5 mmol) 的5 mL乙腈溶液中加入取代苄溴(0.7 mmol), 室温搅拌2 h, TLC监测反应完成后, 减压浓缩, 硅胶匀化, 以二氯甲烷和甲醇为流动相, 经Flash快速柱色谱分离, 得到目标化合物9a和9b。

2 化合物抗新冠病毒活性测定

以HCoV-OC43感染的H460细胞为模型, MPV作为阳性对照药, 在病毒感染(MOI = 1.0) H460细胞(每孔1.5×10⁴个) 的同时将待测化合物不同浓度稀释液和DMSO溶剂对照加入至96孔板(Corning, Inc., Corning, NY) 中, 37 ℃ 5% CO₂培养72 h。采用CCK-8法检测细胞在450 nm波长处的吸光度, 通过比较未感染病毒溶剂对照组、感染病毒溶剂对照组和感染病毒待测化合物组细胞的活力, 判断化合物的抗病毒作用。用Reed-Muench法计算待测化合物抗病毒的EC₅₀值。

3 化合物细胞毒性测定

将H460细胞(每孔1.5×10⁴个) 接种到96孔板上培养过夜, 以MPV作为阳性对照药, 将待测化合物不同浓度稀释液和DMSO溶剂对照加入至96孔板中, 37 ℃ 5% CO₂培养72 h。采用CCK-8法检测细胞在450 nm波长处的吸光度, 通过比较DMSO溶剂对照组和待测化合物组细胞的活力, 判断化合物的细胞毒性。用Reed-Muench法计算待测化合物对细胞的CC₅₀值。

4 表面等离子共振分析(SPR)

SPR实验在BIAcore T200分子互作分析检测系统(GE Healthcare Life Sciences, Piscataway, NJ, USA) 上, 于25 ℃使用CM5芯片进行(GE Healthcare Life Sciences, Piscataway, NJ, USA)。以1×PBS-P+ (GE Healthcare Life Sciences) 为流动相, 在CM5芯片上固定商品化的SARS-CoV-2 NSP12蛋白(华美生物), 再在10 μL·min^-1的流速下使不同浓度的6a和NSP12结合60 s。每次结合完成后, 解离60 s使信号回归基线水平。K_D值用Biacore T200 Evaluation Software v3.0拟合。

5 分子对接

5.1 AutoDock对接

首先使用Chem3D软件MM2 Minimize功能对6a的三维结构进行能量最小化处理, 使用PyMOL对蛋白大分子进行预处理, 仅保留SARS-CoV-2 RdRp (PDB ID: 7ed5) 晶体结构中的NSP12蛋白。使用AutoDock Tools 1.5.6对6a进行加全氢、检测并选择扭转键处理, 对NSP12蛋白进行加全氢处理。在NSP12蛋白的NiRAN催化中心处设置122×126×126的网格盒子, 网格间隔0.338 (x = 122.79, y = 142.42, z = 89.707), 在NSP12催化中心处设置116×126×100的网格盒子, 网格间隔0.299 (x = 131.046, y = 131.312, z = 140.353)。选择Lamarckian遗传算法, 对两个活性腔分别进行对接。选择结合能最低的构象, 用PyMOL 2.4.0进行可视化分析。

5.2 Molegro Virtual Docker对接

按照文献^[23]报道的方法, 使用Molegro Virtual Docker 6.0检测SARS-CoV-2 RdRp (PDB ID: 7ed5) 晶体结构中的NSP12蛋白活性位点, 分别选择NiRAN催化中心和NSP12催化中心为对接腔, 选择MolDock SE算法, 进行对接。选择结合能最低的构象, 用PyMOL 2.4.0进行可视化分析。

5.3 Discovery Studio对接

使用Discovery Studio 4.5软件, 按照文献^[24]报道的方法, 以SARS-CoV-2 RdRp (PDB ID: 7ed5) 晶体结构中的NSP12蛋白为大分子模型, 在PDB晶体结构中记录的活性位点里选择NiRAN催化中心和NSP12催化中心为对接腔, 选用CDocker进行对接。选择结合能最低的构象, 用PyMOL 2.4.0和Discovery Studio 4.5进行可视化分析。

致谢: 目标化合物的核磁共振氢谱、碳谱由中国医学科学院药物研究所分析测试中心测定, 高分辨质谱由中国医学科学院医药生物技术研究所分析测试中心测定。

作者贡献: 梅钰恒负责化学部分、SPR和分子对接实验及初稿的撰写; 李佳玉负责化合物抗病毒活性和细胞毒性的测定; 宋丹青研究员和彭宗根研究员对实验过程中出现的问题及论文撰写提供了有益的指导; 本文的通讯作者李迎红研究员负责实验设计和把关、稿件修改等工作。

利益冲突: 所有作者均声明不存在利益冲突。

基金

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中国医学科学院医学与健康科技创新工程(2021-I2M-1-048)
国家自然科学基金资助项目(82373742)

参考文献

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文献

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参考文献引证文献

排序方式：

[1]

World Health Organization. WHO Coronavirus (COVID-19) [EB/OL]. https://Covid19.Who.Int/ (accessed on 20 July 2023).

[2]

Lin L, Tang F, Xu R, et al. Progress of the development of anti-COVID-19 drugs [J]. Acta Pharm Sin (药学学报), 2023, 58: 39-51.

[3]

Davies NG, Abbott S, Barnard RC, et al. Estimated transmissibility and impact of SARS-CoV-2 lineage B. 1.1. 7 in England [J]. Science, 2021, 372: eabg3055.

[4]

Oude Munnink BB, Sikkema RS, Nieuwenhuijse DF, et al. Transmission of SARS-CoV-2 on mink farms between humans and mink and back to humans [J]. Science, 2021, 371: 172-177.

[5]

Tegally H, Wilkinson E, Lessells RJ, et al. Sixteen novel lineages of SARS-CoV-2 in South Africa [J]. Nat Med, 2021, 27: 440-446.

[6]

Plante JA, Liu Y, Liu J, et al. Spike mutation D614G alters SARS-CoV-2 fitness [J]. Nature, 2021, 592: 116-121.

[7]

Stevens LJ, Pruijssers AJ, Lee HW, et al. Mutations in the SARS-CoV-2 RNA-dependent RNA polymerase confer resistance to remdesivir by distinct mechanisms [J]. Sci Transl Med, 2022, 14: eabo0718.

[8]

Checkmahomed L, Carbonneau J, Du Pont V, et al. In vitro selection of remdesivir-resistant SARS-CoV-2 demonstrates high barrier to resistance [J]. Antimicrob Agents Chemother, 2022, 66: e0019822.

[9]

Torii S, Kim KS, Koseki J, et al. Increased flexibility of the SARS-CoV-2 RNA-binding site causes resistance to remdesivir [J]. PLoS Pathog, 2023, 19: e1011231.

[10]

Gandhi S, Klein J, Robertson AJ, et al. De novo emergence of a remdesivir resistance mutation during treatment of persistent SARS-CoV-2 infection in an immunocompromised patient: a case report [J]. Nat Commun, 2022, 13: 1547.

[11]

Hogan JI, Duerr R, Dimartino D, et al. Remdesivir resistance in transplant recipients with persistent coronavirus disease 2019 [J]. Clin Infect Dis, 2023, 76: 342-345.

[12]

Gao Y, Yan L, Huang Y, et al. Structure of the RNA-dependent RNA polymerase from COVID-19 virus [J]. Science, 2020, 368: 779-782.

[13]

Yin W, Mao C, Luan X, et al. Structural basis for inhibition of the RNA-dependent RNA polymerase from SARS-CoV-2 by remdesivir [J]. Science, 2020, 368: 1499-1504.

[14]

Shannon A, Fattorini V, Sama B, et al. A dual mechanism of action of AT-527 against SARS-CoV-2 polymerase [J]. Nat Commun, 2022, 13: 621.

[15]

Lv X, Zhang M, Yu S, et al. Antiviral and virucidal activities of lycorine on duck tembusu virus in vitro by blocking viral internalization and entry [J]. Poult Sci, 2021, 100: 101404.

[16]

He J, Qi WB, Wang L, et al. Amaryllidaceae alkaloids inhibit nuclear-to-cytoplasmic export of ribonucleoprotein (RNP) complex of highly pathogenic avian influenza virus H5N1 [J]. Influenza Other Respir Viruses, 2013, 7: 922-931.

[17]

Wang P, Li LF, Wang QY, et al. Anti-dengue-virus activity and structure-activity relationship studies of lycorine derivatives [J]. ChemMedChem, 2014, 9: 1522-1533.

[18]

Liang L, Li J, Shen B, et al. The effect of carbonyl on the isomerization of a galanthan ring system and total synthesis of (±)-β-lycorane [J]. Org Biomol Chem, 2021, 19: 2767-2772.

[19]

Chen D, Cai J, Cheng J, et al. Design, synthesis and structure-activity relationship optimization of lycorine derivatives for HCV inhibition [J]. Sci Rep, 2015, 5: 14972.

[20]

Tasker SZ, Cowfer AE, Hergenrother PJ. Preparation of structurally diverse compounds from the natural product lycorine [J]. Org Lett, 2018, 20: 5894-5898.

[21]

Vangeel L, Chiu W, De Jonghe S, et al. Remdesivir, molnupiravir and nirmatrelvir remain active against SARS-CoV-2 Omicron and other variants of concern [J]. Antiviral Res, 2022, 198: 105252.

[22]

Chen H, Lao Z, Xu J, et al. Antiviral activity of lycorine against Zika virus in vivo and in vitro [J]. Virology, 2020, 546: 88-97.

[23]

Jain D, Udhwani T, Sharma S, et al. Design of novel JAK3 inhibitors towards rheumatoid arthritis using molecular docking analysis [J]. Bioinformation, 2019, 15: 68-78.

[24]

Wei W, Zeng Q, Wang Y, et al. Discovery and identification of EIF2AK2 as a direct key target of berberine for anti-inflammatory effects [J]. Acta Pharm Sin B, 2023, 13: 2138-2151.

2024年第59卷第2期

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文章信息

doi: 10.16438/j.0513-4870.2023-1121

接收时间：2023-09-28
首发时间：2025-11-28
出版时间：2024-02-12

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收稿日期：2023-09-28
修回日期：2023-11-07

基金

中国医学科学院医学与健康科技创新工程(2021-I2M-1-048)

国家自然科学基金资助项目(82373742)

作者信息

中国医学科学院、北京协和医学院医药生物技术研究所, 北京 100050

通讯作者:

^*李迎红, Tel: 86-10-63165284, E-mail: liyinghong@imb.pumc.edu.cn

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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Compd.	Yield /%	mp/℃	$ {\left[\alpha \right]}_{\mathrm{D}}^{25\ ^\circ {\rm{C}}} $ (g·100 mL^-1, MeOH)	¹H NMR (600 MHz)	¹³C NMR (151 MHz)	HPLC purity	HR-ESI-MS (m/z)
5a	28%	51-53	-10.00 (0.030)	(CDCl₃) δ 6.86 (s, 1H), 6.59 (s, 1H), 5.96-5.88 (m, 2H), 5.56 (s, 1H), 4.58 (s, 1H), 4.11 (d, J = 14.0 Hz, 1H), 3.92-3.88 (m, 1H), 3.67-3.60 (m, 2H), 3.50 (d, J = 14.0 Hz, 1H), 3.33-3.30 (m, 1H), 2.71 (s, 2H), 2.66-2.57 (m, 2H), 2.36 (q, J = 8.7 Hz, 1H), 1.65-1.52 (m, 2H), 1.45-1.32 (m, 2H), 0.93 (t, J = 7.4 Hz, 3H).	(CDCl₃) δ 146.7, 146.3, 143.5, 130.6, 127.7, 116.0, 107.9, 104.6, 101.0, 79.4, 69.9, 69.0, 61.0, 57.1, 53.9, 41.8, 32.2, 28.8, 19.4, 14.0.	96.9%	[M+H]⁺: C₂₀H₂₆O₄N 344.185 6 344.184 0
5b	20%	114-116	40.91 (0.022)	(CDCl₃) δ 6.86 (s, 1H), 6.60 (s, 1H), 5.96-5.91 (m, 2H), 5.56 (s, 1H), 4.58 (s, 1H), 4.12 (d, J = 14.1 Hz, 1H), 3.92 (s, 1H), 3.72-3.63 (m, 2H), 3.60-3.47 (m, 4H), 3.35-3.32 (m, 1H), 2.71 (s, 2H), 2.66-2.57 (m, 2H), 1.91-1.84 (m, 2H), 1.78-1.73 (m, 2H).	(CDCl₃) δ 146.7, 146.4, 143.7, 130.6, 127.5, 115.8, 107.9, 104.6, 101.1, 79.4, 70.2, 69.1, 61.0, 57.1, 53.9, 45.0, 41.8, 29.5, 28.8, 27.5.	95.2%	[M+H]⁺: C₂₀H₂₅O₄NCl 378.146 7 378.145 2
5c	45%	64-66	-5.56 (0.018)	(DMSO-d₆) δ 6.87 (s, 1H), 6.66 (s, 1H), 5.94 (d, J = 5.3 Hz, 2H), 5.85-5.79 (m, 1H), 5.42-5.39 (m, 1H), 5.01 (dd, J = 17.2, 1.9 Hz, 1H), 4.95 (dd, J = 17.2, 1.9 Hz, 1H), 4.92-4.75 (m, 1H), 4.41 (s, 1H), 4.00 (d, J = 14.1 Hz, 1H), 3.73-3.70 (m, 1H), 3.62 (dt, J = 9.4, 6.2 Hz, 1H), 3.52 (dt, J = 9.4, 6.2 Hz, 1H), 3.43-3.25 (m, 2H), 3.21-3.14 (m, 1H), 2.59 (d, J = 10.4 Hz, 1H), 2.47-2.39 (m, 1H), 2.36 (d, J = 10.4 Hz, 1H), 2.20 (q, J = 8.6 Hz, 1H), 2.07 (q, J = 7.4 Hz, 2H), 1.64-1.53 (m, 2H).	(DMSO-d₆) δ 146.2, 145.7, 143.6, 138.9, 130.1, 129.6, 116.3, 115.4, 107.5, 105.6, 101.1, 80.3, 68.8, 66.9, 61.1, 57.0, 53.6, 41.3, 30.3, 29.2, 28.6.	98.8%	[M+H]⁺: C₂₁H₂₆O₄N 356.185 6 356.184 1
6c	30%	102-104	64.71 (0.017)	(CD₃OD) δ 7.03-6.98 (m, 2H), 5.99 (d, J = 1.2 Hz, 1H), 5.97 (d, J = 1.2 Hz, 1H) 5.74-5.70 (m, 1H), 5.68-5.56 (m, 1H), 5.14 (dt, J = 10.1, 1.4 Hz, 1H), 5.07 (dt, J = 10.1, 1.4 Hz, 1H), 4.77-4.75 (m, 1H), 4.55-4.52 (m, 1H), 4.26-4.15 (m, 3H), 3.92-3.83 (m, 1H), 3.56-3.51 (m, 1H), 3.38-3.33 (m, 1H), 3.08 (d, J = 2.4 Hz, 1H), 3.02-2.97 (m, 2H), 2.85-2.78 (m, 1H), 2.53-2.48 (m, 2H).	(CD₃OD) δ 150.8, 148.5, 137.5, 133.1, 131.6, 124.8, 123.5, 119.6, 109.7, 106.7, 103.1, 73.2, 71.6, 69.9, 64.9, 64.1, 62.6, 39.6, 31.0, 29.1.	98.0%	[M]⁺: C₂₀H₂₄O₄N 342.170 0 342.170 0
6d	20%	150-152	65.22 (0.023)	(CD₃OD) δ 7.00 (s, 1H), 6.98 (s, 1H), 5.99 (d, J = 1.2 Hz, 1H), 5.97 (d, J = 1.2 Hz, 1H), 5.75-5.67 (m, 2H), 5.04-4.87 (m, 2H), 4.73 (d, J = 13.8 Hz, 1H), 4.53-4.51 (m, 1H), 4.24-4.08 (m, 3H), 3.54-3.47 (m, 1H), 3.29 (s, 1H), 3.21-3.10 (m, 1H), 3.01-2.93 (m, 3H), 2.84-2.78 (m, 1H), 2.05-1.93 (m, 2H), 1.86-1.76 (m, 2H).	(CD₃OD) δ 150.8, 148.4, 137.5, 137.4, 131.6, 124.8, 123.4, 117.0, 109.8, 106.7, 103.1, 73.3, 71.5, 69.9, 65.5, 63.9, 62.4, 39.5, 31.2, 31.0, 23.8.	95.5%	[M]⁺: C₂₁H₂₆O₄N 356.185 6 356.184 4
6e	25%	250-252	69.57 (0.023)	(CD₃OD) δ 6.74-6.70 (m, 2H), 5.71-5.66 (m, 2H), 5.45 (d, J = 3.5 Hz, 1H), 4.50-4.44 (m, 2H), 4.27-4.22 (m, 1H), 3.97-3.84 (m, 3H), 2.97-2.92 (m, 4H), 2.77-2.67 (m, 3H), 2.58-2.51 (m, 1H), 1.68-1.53 (m, 2H), 1.46-1.34 (m, 2H).	(CD₃OD) δ 150.8, 148.5, 137.5, 131.6, 124.8, 123.4, 109.8, 106.7, 103.1, 73.3, 71.6, 69.9, 65.0, 64.0, 62.4, 44.6, 39.6, 31.0, 30.2, 22.2.	98.0%	[M]⁺: C₂₀H₂₅O₄NCl 378.146 7 378.144 9
6f	20%	170-172	7.50 (0.040)	(CD₃OD) δ 7.00 (s, 1H), 6.99 (s, 1H), 5.97 (d, J = 3.8 Hz, 2H), 5.74-5.70 (m, 2H), 4.91 (dd, J = 16.5, 2.2 Hz, 1H), 4.85 (dd, J = 10.5, 2.1 Hz, 1H), 4.77-4.72 (m, 3H), 4.52 (s, 2H), 4.23-4.12 (m, 3H), 3.53 (td, J = 12.8, 6.6 Hz, 1H), 3.17 (td, J = 12.4, 4.5 Hz, 1H), 3.01-2.94 (m, 3H), 2.83-2.77 (m, 1H), 1.98-1.95 (m, 2H), 1.75-1.65 (m, 2H), 1.26-1.18 (m, 10H).	(CD₃OD) δ 150.7, 148.4, 140.1, 137.6, 131.6, 124.9, 123.3, 114.7, 109.7, 106.7, 103.1, 73.1, 71.6, 69.9, 66.1, 63.9, 62.4, 39.5, 34.8, 31.0, 30.2, 30.1, 30.0 (2), 27.3, 24.5.	95.2%	[M]⁺: C₂₆H₃₆O₄N 426.263 9 426.262 3
6g	20%	213-215	66.67 (0.030)	(CD₃OD) δ 7.39-7.34 (m, 2H), 7.30 (d, J = 7.9 Hz, 2H), 7.11-6.95 (m, 2H), 6.00 (dd, J = 6.7, 1.3 Hz, 2H), 5.72-5.63 (m, 1H), 4.65-4.48 (m, 2H), 4.27 (d, J = 13.2 Hz, 1H), 4.22-4.09 (m, 5H), 3.42-3.35 (m, 1H), 2.98 (d, J = 11.4 Hz, 1H), 2.74-2.54 (m, 2H), 2.35 (s, 3H).	(CD₃OD) δ 150.8, 148.5, 142.8, 137.6, 134.1 (2), 131.9, 131.3 (2), 126.4, 124.6, 123.4, 110.2, 106.8, 103.2, 72.6, 71.4, 69.9, 68.3, 62.9, 62.4, 39.5, 30.9, 21.3.	96.2%	[M]⁺: C₂₄H₂₆O₄N 392.185 6 392.184 6
6h	50%	214-216	29.03 (0.031)	(CD₃OD) δ 7.84-7.80 (m, 2H), 7.25-7.21 (m, 2H), 7.01 (s, 1H), 6.95 (s, 1H), 6.01 (d, J = 1.2 Hz, 1H), 5.99 (d, J = 1.2 Hz, 1H), 5.72-5.69 (m, 1H), 4.66-4.59 (m, 1H), 4.55-4.53 (m, 1H), 4.31 (d, J = 13.3 Hz, 1H), 4.25 (d, J = 13.3 Hz, 1H), 4.20-4.14 (m, 4H), 3.47-3.40 (m, 1H), 2.97 (d, J = 11.7 Hz, 1H), 2.76-2.69 (m, 2H).	(CD₃OD) δ 150.7, 148.4, 139.9 (2), 137.5, 135.8 (2), 131.7, 129.1, 124.5, 123.6, 110.1, 106.8, 103.2, 98.5, 72.8, 71.4, 69.8, 67.9, 63.6, 62.6, 39.4, 30.8.	96.2%	[M]⁺: C₂₃H₂₃O₄NI 504.066 6 504.065 0
6i	32%	63-65	8.57 (0.035)	(CD₃OD) δ 7.47-7.38 (m, 2H), 7.25-7.11 (m, 2H), 6.81 (s, 1H), 6.56 (s, 1H), 5.89-5.79 (m, 2H), 5.56-5.53 (m, 1H), 4.73-4.64 (m, 2H), 4.59-4.56 (m, 1H), 4.06-4.02 (m, 2H), 3.94-3.92 (m, 1H), 3.49-3.43 (m, 1H), 2.93 (s, 1H), 2.83-2.77 (m, 2H), 2.67-2.57 (m, 2H), 2.56-2.50 (m, 1H), 2.36 (q, J = 8.9 Hz, 1H).	(CD₃OD) δ 148.1, 147.7, 144.7, 139.6, 130.4, 130.3 (2), 129.6, 122.8, 122.0 (2), 121.1, 117.3, 108.2, 106.1, 102.3, 80.9, 71.7, 68.9, 62.4, 57.8, 54.7, 42.2, 29.3.	97.5%	[M]⁺: C₂₄H₂₃O₅NF₃ 462.152 3 462.150 3
6j	44%	210-212	46.43 (0.028)	(CD₃OD) δ 7.68 (d, J = 1.7 Hz, 1H), 7.62 (d, J = 1.7 Hz, 2H), 6.98 (s, 1H), 6.70 (s, 1H), 6.00–5.94 (m, 2H), 5.75-5.71 (m, 1H), 4.69 (d, J = 14.1 Hz, 1H), 4.53-4.51 (m, 1H), 4.44-4.39 (m, 2H), 4.31-4.18 (m, 4H), 3.66-3.53 (m, 1H), 2.94 (d, J = 11.5 Hz, 1H), 2.82-2.78 (m, 2H).	(CD₃OD) δ 150.6, 148.3, 137.4, 135.8 (3), 135.5, 133.6, 131.5, 124.6, 123.7 (2), 109.6, 106.6, 103.2, 72.4, 71.4, 69.8, 67.7, 65.7, 63.3, 54.2, 30.8.	98.0%	[M]⁺: C₂₃H₂₂O₄NBr₂ 533.991 0 533.989 3 (⁷⁹Br+⁷⁹Br) 535.987 4 (⁷⁹Br+⁸¹Br) 537.985 2 (⁸¹Br+⁸¹Br)
9a	24%	194-196	17.39 (0.023)	(CD₃OD) δ 7.84-7.81 (m, 2H), 7.26-7.20 (m, 2H), 6.93 (s, 1H), 6.63 (s, 1H), 6.00-5.97 (m, 2H), 5.72-5.70 (m, 1H), 4.70-4.66 (m, 1H), 4.46-4.35 (m, 2H), 4.24-4.20 (m, 3H), 3.55-3.45 (m, 2H), 3.26 (s, 1H), 3.24 (s, 1H), 2.94-2.80 (m, 2H), 2.06 (s, 3H).	(CD₃OD) δ 170.1, 149.4, 147.3, 138.7, 138.4, 136.1, 134.4, 134.2, 128.8, 127.6, 123.1, 122.1, 109.1, 103.9, 101.9, 97.1, 71.0, 70.3, 66.8, 66.7, 62.7, 60.7, 36.8, 29.4, 19.3.	95.1%	[M]⁺: C₂₅H₂₅O₅NI 546.077 2 546.074 7
9b	24%	208-210	36.00 (0.025)	(CD₃OD) δ 7.62 (d, J = 1.8 Hz, 2H), 7.54 (d, J = 1.8 Hz, 1H), 6.53 (s, 1H), 6.52 (s, 1H), 5.95-5.90 (m, 2H), 5.68-5.66 (m, 1H), 4.87 (d, J = 13.9 Hz, 1H), 4.73 (d, J = 13.3 Hz, 1H), 4.61-4.50 (m, 2H), 4.46 (d, J = 11.6 Hz, 1H), 4.23-4.21 (m, 1H), 4.07-3.93 (m, 1H), 3.80-3.73 (m, 1H), 3.60-3.49 (m, 1H), 3.23-3.18 (m, 1H), 3.02-2.95 (m, 1H), 2.93-2.86 (m, 1H), 2.13 (s, 3H).	(CD₃OD) δ 171.5, 150.5, 148.4, 137.7, 137.2, 135.8 (2), 135.7, 133.7, 129.6, 123.6 (2), 109.8, 108.5, 105.0, 103.3, 71.8, 68.7, 68.3, 63.5, 62.1, 60.8, 53.3, 38.2, 30.9.	97.0%	[M]⁺: C₂₅H₂₄O₅NBr₂ 576.001 6 575.999 0 (⁷⁹Br+⁷⁹Br) 577.996 6 (⁷⁹Br+⁸¹Br) 579.994 8 (⁸¹Br+⁸¹Br)

Compd.

Yield /%

mp/℃

$ {\left[\alpha \right]}_{\mathrm{D}}^{25\ ^\circ {\rm{C}}} $

(g·100 mL^-1, MeOH)

¹H NMR (600 MHz)

¹³C NMR (151 MHz)

HPLC purity

HR-ESI-MS (m/z)

Formula Calcd. Found

28%

51-53

-10.00
(0.030)

(CDCl₃) δ 146.7, 146.3, 143.5, 130.6, 127.7, 116.0, 107.9, 104.6, 101.0, 79.4, 69.9, 69.0, 61.0, 57.1, 53.9, 41.8, 32.2, 28.8, 19.4, 14.0.

96.9%

[M+H]⁺:
C₂₀H₂₆O₄N
344.185 6
344.184 0

20%

114-116

40.91
(0.022)

(CDCl₃) δ 146.7, 146.4, 143.7, 130.6, 127.5, 115.8, 107.9, 104.6, 101.1, 79.4, 70.2, 69.1, 61.0, 57.1, 53.9, 45.0, 41.8, 29.5, 28.8, 27.5.

95.2%

[M+H]⁺:
C₂₀H₂₅O₄NCl 378.146 7
378.145 2

45%

64-66

-5.56
(0.018)

(DMSO-d₆) δ 146.2, 145.7, 143.6, 138.9, 130.1, 129.6, 116.3, 115.4, 107.5, 105.6, 101.1, 80.3, 68.8, 66.9, 61.1, 57.0, 53.6, 41.3, 30.3, 29.2, 28.6.

98.8%

[M+H]⁺:
C₂₁H₂₆O₄N
356.185 6
356.184 1

30%

102-104

64.71
(0.017)

(CD₃OD) δ 150.8, 148.5, 137.5, 133.1, 131.6, 124.8, 123.5, 119.6, 109.7, 106.7, 103.1, 73.2, 71.6, 69.9, 64.9, 64.1, 62.6, 39.6, 31.0, 29.1.

98.0%

[M]⁺:
C₂₀H₂₄O₄N
342.170 0
342.170 0

20%

150-152

65.22
(0.023)

(CD₃OD) δ 150.8, 148.4, 137.5, 137.4, 131.6, 124.8, 123.4, 117.0, 109.8, 106.7, 103.1, 73.3, 71.5, 69.9, 65.5, 63.9, 62.4, 39.5, 31.2, 31.0, 23.8.

95.5%

[M]⁺:
C₂₁H₂₆O₄N
356.185 6
356.184 4

25%

250-252

69.57
(0.023)

(CD₃OD) δ 150.8, 148.5, 137.5, 131.6, 124.8, 123.4, 109.8, 106.7, 103.1, 73.3, 71.6, 69.9, 65.0, 64.0, 62.4, 44.6, 39.6, 31.0, 30.2, 22.2.

98.0%

[M]⁺:
C₂₀H₂₅O₄NCl 378.146 7
378.144 9

20%

170-172

7.50
(0.040)

(CD₃OD) δ 150.7, 148.4, 140.1, 137.6, 131.6, 124.9, 123.3, 114.7, 109.7, 106.7, 103.1, 73.1, 71.6, 69.9, 66.1, 63.9, 62.4, 39.5, 34.8, 31.0, 30.2, 30.1, 30.0 (2), 27.3, 24.5.

95.2%

[M]⁺:
C₂₆H₃₆O₄N
426.263 9
426.262 3

20%

213-215

66.67
(0.030)

(CD₃OD) δ 150.8, 148.5, 142.8, 137.6, 134.1 (2), 131.9, 131.3 (2), 126.4, 124.6, 123.4, 110.2, 106.8, 103.2, 72.6, 71.4, 69.9, 68.3, 62.9, 62.4, 39.5, 30.9, 21.3.

96.2%

[M]⁺:
C₂₄H₂₆O₄N
392.185 6
392.184 6

50%

214-216

29.03
(0.031)

(CD₃OD) δ 150.7, 148.4, 139.9 (2), 137.5, 135.8 (2), 131.7, 129.1, 124.5, 123.6, 110.1, 106.8, 103.2, 98.5, 72.8, 71.4, 69.8, 67.9, 63.6, 62.6, 39.4, 30.8.

96.2%

[M]⁺:
C₂₃H₂₃O₄NI 504.066 6
504.065 0

32%

63-65

8.57
(0.035)

(CD₃OD) δ 148.1, 147.7, 144.7, 139.6, 130.4, 130.3 (2), 129.6, 122.8, 122.0 (2), 121.1, 117.3, 108.2, 106.1, 102.3, 80.9, 71.7, 68.9, 62.4, 57.8, 54.7, 42.2, 29.3.

97.5%

[M]⁺:
C₂₄H₂₃O₅NF₃ 462.152 3
462.150 3

44%

210-212

46.43
(0.028)

(CD₃OD) δ 150.6, 148.3, 137.4, 135.8 (3), 135.5, 133.6, 131.5, 124.6, 123.7 (2), 109.6, 106.6, 103.2, 72.4, 71.4, 69.8, 67.7, 65.7, 63.3, 54.2, 30.8.

98.0%

[M]⁺:
C₂₃H₂₂O₄NBr₂ 533.991 0
533.989 3 (⁷⁹Br+⁷⁹Br)
535.987 4 (⁷⁹Br+⁸¹Br)
537.985 2 (⁸¹Br+⁸¹Br)

24%

194-196

17.39
(0.023)

(CD₃OD) δ 170.1, 149.4, 147.3, 138.7, 138.4, 136.1, 134.4, 134.2, 128.8, 127.6, 123.1, 122.1, 109.1, 103.9, 101.9, 97.1, 71.0, 70.3, 66.8, 66.7, 62.7, 60.7, 36.8, 29.4, 19.3.

95.1%

[M]⁺:
C₂₅H₂₅O₅NI 546.077 2
546.074 7

24%

208-210

36.00
(0.025)

(CD₃OD) δ 171.5, 150.5, 148.4, 137.7, 137.2, 135.8 (2), 135.7, 133.7, 129.6, 123.6 (2), 109.8, 108.5, 105.0, 103.3, 71.8, 68.7, 68.3, 63.5, 62.1, 60.8, 53.3, 38.2, 30.9.

97.0%

[M]⁺:
C₂₅H₂₄O₅NBr₂ 576.001 6
575.999 0 (⁷⁹Br+⁷⁹Br)
577.996 6 (⁷⁹Br+⁸¹Br)
579.994 8 (⁸¹Br+⁸¹Br)


3	/	/	/	＞50.00	＞200.00	N/A
4	/	/	/	21.81 ± 2.72	154.66 ± 4.15	7.09
5a	H		/	＞50.00	＞200.00	N/A
5b	H		/	＞50.00	83.43 ± 2.80	＜1.67
5c	H		/	＞50.00	＞200.00	N/A
6a	H	H		2.36 ± 0.24	38.98 ± 1.04	16.52
6b	H	H		35.43 ± 9.92	＞200.00	＞5.64
6c	H	H		＞50.00	＞200.00	N/A
6d	H	H		＞50.00	＞200.00	N/A
6e	H	H		33.79 ± 6.55	＞200.00	＞5.92
6f	H	H		3.90 ± 3.00	＞200.00	＞51.28
6g	H	H		＞50.00	65.83 ± 1.94	＜1.32
6h	H	H		23.10 ± 0.40	＞200.00	＞8.66
6i	H	H		6.00 ± 0.80	21.50 ± 1.60	3.58
6j	H	H		5.59 ± 0.63	＞200.00	＞35.78
7	/	/	/	＞50.00	＞200.00	N/A
9a		H		36.50 ± 0.10	＞200.00	＞5.48
9b		H		33.90 ± 1.90	＞200.00	＞5.90
1	/	/	/	0.69 ± 0.06	6.74 ± 0.34	9.77
MPV	/	/	/	0.21 ± 0.01	78.58 ± 26.24	374.19

Compd.

R₁

R₂

R₃

EC₅₀/μmol·L^{-1 a}

CC₅₀/μmol·L^{-1 b}

SI^c

＞50.00

＞200.00

N/A

21.81 ± 2.72

154.66 ± 4.15

7.09

＞50.00

＞200.00

N/A

＞50.00

83.43 ± 2.80

＜1.67

＞50.00

＞200.00

N/A

2.36 ± 0.24

38.98 ± 1.04

16.52

35.43 ± 9.92

＞200.00

＞5.64

＞50.00

＞200.00

N/A

＞50.00

＞200.00

N/A

33.79 ± 6.55

＞200.00

＞5.92

3.90 ± 3.00

＞200.00

＞51.28

＞50.00

65.83 ± 1.94

＜1.32

23.10 ± 0.40

＞200.00

＞8.66

6.00 ± 0.80

21.50 ± 1.60

3.58

5.59 ± 0.63

＞200.00

＞35.78

＞50.00

＞200.00

N/A

36.50 ± 0.10

＞200.00

＞5.48

33.90 ± 1.90

＞200.00

＞5.90

0.69 ± 0.06

6.74 ± 0.34

9.77

MPV

0.21 ± 0.01

78.58 ± 26.24

374.19

Item	AutoDock		Discovery Studio		MVD
6a-NiRAN catalytic center	-7.33		-27.65		-98.39
6a-NSP12 catalytic center	-6.02		-21.57		-97.94

Item

AutoDock

Discovery Studio

MVD

Binding energy /kcal·mol^-1

CDocker interaction energy/kcal·mol^-1

MolDock score

6a-NiRAN catalytic center

-7.33

-27.65

-98.39

6a-NSP12 catalytic center

-6.02

-21.57

-97.94