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Supporting Information for
A convenient Michael addition of imidazoles to acrylates catalyzed
by Lipozyme TL IM from Thermomyces lanuginosus in a
continuous‐flow microreactor
Li‐Hua Du *a, Zhen Donga, Rui‐Jie Longa, Ping‐Feng Chena, Miao Xuea and Xi‐Ping Luo*b
aCollege of Pharmaceutical Science, ZheJiang University of Technology, Zhejiang,Hangzhou, 310014, People’s Republic of China
b College of Science, Zhejiang A&F University, Zhejiang, Hangzhou, 311300, China ∗Corresponding author. Fax: +86 571 88320903. E‐mail: [email protected]; [email protected].
Materials
Unless otherwise stated, all chemicals were obtained from commercial sources and used without further
purification. Lipozyme TL IM from Thermomyces lanuginosus was purchased from Novo Nordisk.
Imidazole, acrylates and all other chemicals were of the highest purity commercially available and
without further purification. Harvard Apparatus PHD 2000 syringe pumps were purchased from Harvard
apparatus.
Experimental setup
The enzymatic Michael addition of imidazole derivatives with acrylates is described in Fig. 1. The micro‐flow
reactor was composed of a syringe pump (Harvard Apparatus PHD 2000), reactant injector, Y‐shaped mixer,
microchannel reactor and product collector. Syringe pumps were used to deliver reagents from reactant injectors
to Y‐shaped mixer and then to the microchannel reactor. Reagent feed 1 (10 mL) with the imidazole derivatives in
DMSO solution and reagent feed 2 (10 mL) with acrylates in DMSO solution were fully mixed in Y‐shaped mixer.
Lipozyme TL IM (catalyst reactivity: 250 IUN.g‐1) were filled in microchannel reactor made of a PFA reactor coil
(inner diameter ID = 1.8 mm, length = 1 m). The microchannel reactor with catalyst in it was submerged into a
thermostatic water bath to control the temperature. Feed 1 and 2 were mixed together at a flow rate of 7.3 µL
min‐1 in a Y‐mixer at 45 °C and the resulting stream (14.6 µL min‐1) was connected to a sample vial which was
used to collect the final mixture.
Fig. 1 Experimental setup for Michael addition of imidazole derivatives with acrylates catalyzed by lipase TL IM in a continuous flow microreactor.
Electronic Supplementary Material (ESI) for Organic & Biomolecular Chemistry.This journal is © The Royal Society of Chemistry 2018
General Procedure for Michael addition reaction in Continuous Flow
Microreactors
Method B: 1.0 mmol of the imidazole derivatives was dissolved in 10 mL DMSO (feed 1, ~0.1 M) and 4.0
mmol acrylates were dissolved in 10 mL DMSO (feed 2; ~0.4 M). Lipozyme TL IM (0.87 g) were filled in PFA
reactor coil (inner diameter ID= 1.8 mm, length = 1m. We filled the silica gel‐loaded enzyme granules in a
pipe by physical filling, and then filled the enzyme evenly with an ultrasonic oscillator.). Streams 1 and 2
were mixed together at a flow rate of. 7.3 µL min‐1 in a Y‐mixer at 45 °C and the resulting stream (14.6 µL
min‐1) was connected to a sample vial which was used to collect the final mixture. The final mixture was
then evaporated, and the residue was submitted to column chromatography on silica gel (200‐300 mesh).
The products were eluted with a gradient of petroleum ether /ethyl acetate (4:1, by vol). The purification
was monitored by TLC. The fractions containing the main products were pooled, the solvent evaporated,
and the residue analyzed by 1H NMR, 13C NMR and ESI‐MS.
General Procedure for Michael addition reaction under Shaker Conditions.
Method A: imidazole derivatives (0.25 mmol) and acrylates (1.0 mmol) were added to 5 mL DMSO. The
biocatalyst lipozyme TL IM (40 mg/mL, 0.20 g) was then added and the suspension maintained at 45 °C
for 24 h under Shaker Conditions. The mixture was cooled and filtered. Then evaporated under reduced
pressure and the residue was submitted to column chromatography on silica gel (200– 300 mesh). The
products were eluted with a gradient of normal petroleum ether/ethyl acetate (4:1, by vol). The
purification was monitored by TLC. The fractions containing the main products were pooled, the solvent
evaporated, and the residue analyzed by 1H NMR, 13C NMR and ESI‐MS.
Thin‐Layer Chromatography
Analytical TLC was performed on silica gel 60 plates (Merck) using petroleum ether /ethyl acetate(2:1, by
vol) as developing solvent. Spots were detected by ultraviolet irradiation at 254 nm.
High Performance Liquid Chromatography (HPLC)
The reaction was monitored by HPLC analysis using a Shim‐Pack VP‐ODS column (4.6×150 mm) and a UV
detector (285 nm). MeOH/NaH2PO4 solution (7.5×10‐3 M) (25:75) was used as eluant at 1.0 mL min‐1 for
imidazole and acrylates. MeOH/H2O solution (30:70) was used as eluant at 1.0 mL min‐1 for
nitroimidazole and acrylates. The yield was defined as the ratio between the molar concentration of
N‐substituted imidazole derivatives and the initial molar concentration of the imidazole derivatives used.
In order to examine the reproducibility of the method, we repeated the reaction five times, the result
are illustrate in Figure 2.
Figure 2.The reproducibility of the reaction on the conversion of 3‐(4‐Nitroimidazol‐1‐yl)‐propionic acid
methyl ester by Lipozyme TL IM in a continuous flow microreactor.
The reaction time is calculated in two ways:
(1) Actual reaction time: We use the timer calculated the reaction time: the start time of the reaction
begin from the two reaction solution first contact with the catalyst in microreactor to the first
droplet of product flow out of the tube of microreactor.
(2) Theoretical reaction time: There is a display data on the microreactor injection pump and the data
will increase with the continuous injection. For example, we choose a flow rate (0.0073 mL/min),
when the two reaction solution first contact with the catalyst in microreactor, a data display( 0.0670
mL ), another data is got at the time the first droplet of product flow out of the tube of microreactor
( 0.3223 mL ). So the reaction time is: (0.3223‐0.0670)/0.0073= 34.9 min.
We have five flow rates: 0.0127 mL/min, 0.0102 mL/min, 0.0085 mL/min, 0.0073 mL/min, 0.0064
mL/min. From the Table 1, we can see that no matter which flow rate we test, the two data on the
microreactor is almost the same. The Actual time and the Theoretical time are almost the same as
well.
Table 1
First data (mL)
Second data
(mL)
Flow rate (mL/min)
Actual time (min)
way 1
Theoretical time
(min) way 2
0.0673 0.3220 0.0127 20 20.05
0.0672 0.3220 0.0102 25 24.98
0.0673 0.3222 0.0085 30 29.98
0.0670 0.3223 0.0073 35 34.97
0.0671 0.3225 0.0064 40 39.91
Figure 3 Mechanism of lipase catalyzed Michael addition.
Nuclear magnetic resonance (NMR) analysis
After purification of the synthesized products by column chromatography, the chemical structures of
imidazoles were determined by 1H NMR, 13C NMR and ESI‐MS.
3‐(4‐Nitroimidazol‐1‐yl)‐propionic acid methyl ester (3a)[1]. White solid, mp: 109°C; 1H NMR (500 MHz,
CDCl3): δ = 7.84 (d, 1H, J = 1.1 Hz, N=CHN), 7.52 (d, 1H, J = 1.1 Hz, NCH=C), 4.36 (t, 2H, J = 6.1 Hz, NCH2), 3.73
(s, 3H, OCH3), 2.87 (t, 2H, J = 6.1 Hz, CH2C=O); 13C NMR (125 MHz, CDCl3): δ = 170.38 (C=O), 148.71
(C4),136.34 (C2), 119.33 (C5), 52.46 (OCH3), 43.55 (NCH2), 35.08 (CH2C=O). ESI‐MS (m/z): 200 (M+1); IR (KBr):
3079, 3042, 1729, 1526, 1485, 1336, 1204, 1179, 1153,991, 822, 760, 656 cm‐1.
3‐(4‐Nitroimidazol‐1‐yl)‐propionic acid ethyl ester (3b)[1]. White solid, mp: 36°C; 1H NMR (500 MHz, CDCl3):
δ = 7.88 (d, 1H, J = 1.1 Hz, N=CHN), 7.57 (d, 1H, J = 1.1 Hz, NCH=C), 4.37 (t, 2H, J = 6.1 Hz, NCH2), 4.14 (q, 2H,
J = 7.1 Hz, OCH2), 2.85 (t, 2H, J = 6.1 Hz, CH2C=O), 1.23 (t, 3H, J = 7.1 Hz, CH2CH3); 13C NMR(125 MHz, CDCl3):
δ = 169.98 (C=O), 147.83 (C4), 136.46 (C2), 119.64 (C5), 61.48 (OCH2), 43.63 (NCH2), 35.18 (CH2C=O), 13.98
(CH2CH3); ESI‐MS (m/z): 214 (M+1); IR (KBr): 3079, 2979, 1732, 1525, 1486, 1455,1339, 1201, 990, 880, 824,
654 cm‐1.
3‐(4‐Nitroimidazol‐1‐yl)‐propionic acid butyl ester (3c)[1]. White solid, mp: 48°C; 1H NMR (500 MHz, CDCl3):
δ = 7.88 (d, 1H, J = 1.1 Hz, N=CHN), 7.58 (d, 1H, J = 1.1 Hz, NCH=C), 4.37 (t, 2H, J = 6.1 Hz, NCH2), 4.09 (t, 2H, J
= 6.6 Hz, OCH2), 2.86 (t, 2H, J = 6.1 Hz, CH2C=O), 1.57 (m, 2H, CH2CH2CH2), 1.31 (m, 2H, CH2CH2CH3), 0.89 (t,
3H, J = 7.4 Hz, CH2CH3); 13C NMR (125 MHz, CDCl3): δ = 170.08 (C=O), 147.82 (C4), 136.45 (C2), 119.62 (C5),
65.36 (OCH2), 43.66 (NCH2), 35.16 (CH2C=O), 30.37 (CH2CH2CH2), 18.92 (CH2CH3), 13.50 (CH2CH3); ESI‐MS
(m/z): 242 (M+1); IR (KBr): 2963, 1727, 1524, 1488, 1338, 1296, 1205, 990, 880, 823, 655 cm‐1.
3‐(4‐Nitroimidazol‐1‐yl)‐propionic acid tert‐butyl ester (3d)[1]. White solid, mp: 50°C; 1H NMR (500 MHz,
CDCl3): δ = 7.83 (d, 1H, J = 1.1 Hz, N=CHN), 7.51 (d, 1H, J = 1.1 Hz, NCH=C), 4.31 (t, 2H, J = 6.1 Hz, NCH2), 2.76
(t, 2H, J = 6.1 Hz, CH2C=O), 1.43 (s, 9H, C(CH3)3); 13C NMR (125 MHz, CDCl3): δ = 169.12 (C=O), 147.71 (C4),
136.34 (C2), 119.38 (C5), 82.51 (OC(CH3)3), 43.83 (NCH2), 36.37 (O=CCH2), 27.97 (C(CH3)3); ESI‐MS (m/z): 242
(M+1); IR (KBr): 2976, 1728, 1526, 1486, 1342, 1167, 991, 882, 824, 759, 654 cm‐1.
3‐(2‐Methyl‐4‐nitroimidazol‐1‐yl)‐propionic acid methyl ester (3f)[2]. Syrupy liquid; 1H NMR (500 MHz,
CDCl3): δ = 7.75 (s, 1H, NCH=C), 4.23 (t, 2H, J = 6.4 Hz, NCH2), 3.66 (s, 3H, OCH3), 2.81 (t, 2H, J = 6.4 Hz,
CH2C=O), 2.43 (s, 3H, N=CCH3); 13C NMR (125 MHz, CDCl3): δ = 170.21 (C=O), 146.35 (C4), 144.81 (C2), 119.82
(C5), 52.20 (OCH3), 42.24 (NCH2), 34.25 (CH2C=O), 12.87 (N=CCH3); ESI‐MS (m/z): 214(M+1); IR (KBr): 3104,
1737, 1540, 1496, 1329, 1212, 1145, 997, 848, 826, 758, 677, 561 cm‐1.
3‐(2‐Methyl‐4‐nitroimidazole‐1‐yl)‐propionic acid Ethyl ester (3g)[2]. Syrupy liquid; 1H NMR (500 MHz,
CDCl3): δ = 7.74 (s, 1H, NCH=C), 4.20 (t, 2H, J = 6.4 Hz, NCH2), 4.07 (q, 2H, J = 7.1 Hz, OCH2), 2.77 (t, 2H, J =
6.4 Hz, O=CCH2), 2.40 (s, 3H, N=CCH3), 1.16 (t, 3H, J = 7.1 Hz, CH2CH3). 13CNMR (125 MHz, CDCl3): δ = 169.71
(C=O), 146.21 (C4), 144.77 (C2), 119.87 (C5), 61.24 (OCH2), 42.23 (NCH2), 34.40 (O =CCH2), 13.82 (N=CCH3),
12.79 (CH2CH3). ESI–MS (m/z): 228 (M + 1); IR (KBr): 3142, 2991, 1729, 1537, 1500, 1408, 1322, 1292, 1202,
1024, 945, 826, 805, 754, 556 cm‐1.
3‐(2‐Methyl‐4‐nitroimidazole‐1‐yl)‐propionic acid butyl ester (3h)[2]. Hazel columnar crystal, mp: 41°C; 1H
NMR (500 MHz, CDCl3): δ = 7.69 (s, 1H, NCH=C), 4.14 (t, 2H, J = 6.4 Hz, NCH2), 3.92 (t, 2H, J = 6.7 Hz, OCH2),
2.71 (t, 2H, J = 6.4 Hz, O=CCH2), 2.31 (s, 3H, N=CCH3), 1.41 (m, 2H, CH2CH2CH2), 1.14 (m, 2H, CH2CH3), 0.72 (t,
3H, J = 7.4 Hz, CH2CH3). 13CNMR (125 MHz, CDCl3): δ = 169.95 (O=C), 146.08 (C4), 144.91 (C2), 120.11 (C5),
65.02 (OCH2), 42.32 (NCH2), 34.37 (O=CCH2), 30.24 (OCH2CH2), 18.77 (CH2CH3), 13.35 (N=CCH3), 12.76
(CH2CH3). ESI–MS (m/z): 256 (M + 1); IR (KBr): 3102, 2963, 2937, 1729, 1542, 1505, 1415, 1325, 1292, 1209,
1146, 999, 826, 758, 562 cm‐1.
3‐(2‐Methyl‐4‐nitroimidazole‐1‐yl)‐propionic acid tert‐butyl ester (3i). Hazel columnar crystal, mp: 45°C; 1H
NMR (500 MHz, CDCl3): δ = 7.75(s, 1H, NCH=C), 4.18 (t, 2H, J = 6.4 Hz, NCH2), 2.71 (t, 2H, J = 6.4 Hz, O=CCH2),
2.45 (s, 3H, N=CCH3), 1.40 (s, 9H, C(CH3)3). 13C NMR (125 MHz, CDCl3): δ = 168.95 (O=C), 146.11 (C4), 144.77
(C2), 119.74 (C5), 82.35 (OC(CH3)3), 42.56 (NCH2), 35.70 (O=CCH2 ), 27.90 (OC(CH3)3), 13.01 (N=CCH3). ESI–MS
(m/z): 256 (M + 1); IR (KBr): 3109, 2979, 1727, 1537, 1499, 1415, 1318, 1290, 1143, 999, 828, 758 cm‐1.
3‐(6‐nitro‐benzimidazole‐1‐yl)‐propionic acid methyl ester and 3‐(5‐nitro‐benzimidazole‐1‐yl)‐propionic
acid methyl ester (3p). Yellow oil; 1H NMR (500 MHz, CDCl3): δ = 8.71 (d, 1H, J = 2.0 Hz, Ar‐H), 8.40 (d, 1H, J =
2.0 Hz, Ar‐H), 8.27‐8.20 (m, 4H, Ar‐H), 7.87 (d, 1H, J = 8.9 Hz, Ar‐H), 7.50 (d, 1H, J = 8.9 Hz, Ar‐H), 4.60 (m, 4H,
NCH2), 3.69 (s, 6H, OCH3), 2.94 (m, 4H, O=CCH2); 13C NMR (125 MHz, CDCl3): δ = 170.71, 147.97, 146.78,
143.99, 143.90, 143.09, 137.54, 132.77, 120.66, 118.96, 118.19, 117.25, 109.45, 106.47, 52.32, 40.82, 40.77,
34.09. ESI–MS (m/z): 250 (M + 1); IR (KBr): 2957, 1726, 1519, 1435, 1340, 1205, 1176, 980, 935, 788, 739
cm‐1.
3‐(6‐nitro‐benzimidazole‐1‐yl)‐propionic acid Ethyl ester and 3‐(5‐nitro‐benzimidazole‐1‐yl)‐propionic acid
Ethyl ester (3q). yellow oil; 1H NMR (500 MHz, CDCl3): δ = 8.52 (d, 1H, J = 2.1 Hz, Ar‐H), 8.32 (d, 1H, J = 2.1 Hz,
Ar‐H), 8.24 (s, 1H, Ar‐H), 8.16 (s, 1H, Ar‐H), 8.06 (m, 2H, Ar‐H), 7.72 (d, 1H, J = 8.9 Hz, Ar‐H), 7.45 (d, 1H, J =
8.9 Hz, Ar‐H) 4.54 (m, 4H, NCH2), 4.02 (m, 4H, OCH2), 2.87 (m, 4H, O=CCH2), 1.10 (m, 6H, OCH2CH3); 13C NMR
(125 MHz, CDCl3): δ = 170.20, 147.59, 146.68, 145.78, 144.28, 144.05, 141.75, 137.25, 132.47, 119.98,
119.16, 118.73, 116.67, 109.90, 106.99, 61.45, 41.20, 34.25, 13.99. ESI–MS (m/z): 264 (M + 1); IR (KBr): 3095,
2976, 1716, 1510, 1337, 1295, 1199, 1167, 1061, 1013, 889, 824, 740 cm‐1.
3‐(6‐nitro‐benzimidazole‐1‐yl)‐propionic acid Butyl ester and 3‐(5‐nitro‐benzimidazole‐1‐yl)‐propionic acid
Butyl ester (3r)[3]. Yellow oil; 1H NMR (500 MHz, CDCl3): δ = 8.89 (s, 1H, Ar‐H), 8.75 (d, 1H, J = 2.0 Hz, Ar‐H),
8.63 (s, 1H, Ar‐H), 8.51 (d, 1H, J = 2.0 Hz, Ar‐H), 8.30 (m, 2H, Ar‐H), 7.96 (d, 1H, J = 8.9 Hz, Ar‐H), 7.60 (d, 1H, J
= 8.9 Hz, Ar‐H), 4.68 (m, 4H, NCH2), 4.07 (m, 4H, OCH2), 2.98 (m, 4H, O=CCH2), 1.67 (m, 4H, OCH2CH2), 1.28
(m, 4H, CH2CH3), 0.90 (t, J = 7.2 Hz, 6H, CH2CH3); 13C NMR (125 MHz, CDCl3): δ = 170.36, 147.35, 146.57,
144.30, 140.94, 137.04, 132.28, 119.71, 119.44, 119.21, 116.49, 110.17, 107.28, 65.43, 41.39, 34.25, 30.40,
18.97, 13.58. ESI–MS (m/z): 292 (M + 1); IR (KBr): 3436, 3093, 1725, 1518, 1435, 1384, 1341, 1206, 1176,
788, 739 cm‐1.
N
NO
OO2N
N
N O
O
O2N
3‐(6‐nitro‐benzimidazole‐1‐yl)‐propionic acid tert‐butyl ester and 3‐(5‐nitro‐benzimidazole‐1‐yl)‐propionic
acid tert‐butyl ester (3s). Yellow oil; 1H NMR (500 MHz, CDCl3): δ = 8.72 (d, 1H, J = 2.0 Hz, Ar‐H), 8.42 (d, 1H,
J = 2.0 Hz, Ar‐H), 8.29‐8.21 (m, 4H, Ar‐H), 7.88 (d, 1H, J = 8.9 Hz, Ar‐H), 7.51 (d, 1H, J = 8.9 Hz, Ar‐H), 4.55 (m,
4H, NCH2), 2.84 (m, 4H, O=CCH2), 1.39 (s, 18H, OC(CH3)3); 13C NMR (125 MHz, CDCl3): δ = 169.48, 147.91,
146.77, 143.98, 142.94, 137.57, 132.78, 120.53, 118.90, 118.21, 117.17, 109.61, 106.65, 82.28, 82.25,
41.09, 41.03, 35.46, 35.39, 27.96. ESI–MS (m/z): 292 (M + 1); IR (KBr): 2978, 1725, 1517, 1371, 1345, 1231,
1157, 1061, 884, 843, 796, 741 cm‐1.
1H‐NMR of compound 3a
13C‐NMR of compound 3a
1H‐NMR of compound 3b
13C‐NMR of compound 3b
1H‐NMR of compound 3c
13C‐NMR of compound 3c
1H‐NMR of compound 3d
13C‐NMR of compound 3d
1H‐NMR of compound 3f
N
NO2N
O
O
13C‐NMR of compound 3f
1H‐NMR of compound 3g
13C‐NMR of compound 3g
1H‐NMR of compound 3h
13C‐NMR of compound 3h
1H‐NMR of compound 3i
13C‐NMR of compound 3i
1H‐NMR of compound 3p
13C‐NMR of compound 3p
N
NO
O
N
N O
OO2N
O2N
1H‐NMR of compound 3q
13C‐NMR of compound 3q
1H‐NMR of compound 3r
13C‐NMR of compound 3r
1H‐NMR of compound 3s
13C‐NMR of compound 3s
The Mass spectrum of compound
compound 3a
compound 3b
compound 3c
compound 3d
compound 3f
compound 3g
compound 3h
compound 3i
compound 3i
compound 3p
compound 3q
compound 3r
compound 3s
The IR spectra of compound
compound 3a
656.
12759.
618
22.
70
83
9.9
888
7.10
932.
7796
5.81
990.
6810
43.9
810
57.5
3
1153
.31
1178
.74
1204
.54
123
2.2
91
262
.37
1295
.33
13
36.2
31
365
.17
138
8.6
814
06.5
91
42
3.0
514
41.8
114
84.8
81
526
.38
1544
.02
172
8.4
8
2957
.81
30
42.2
53
079
.28
31
37.1
9
34
49.8
0
40
45
50
55
60
65
70
75
80
85
90
95
100
%T
500 1000 1500 2000 2500 3000 3500 4000
Wavenumbers (cm-1)
compound 3b
compound 3c
653
.89
759.
20
823
.58
854
.33
879
.98
941.
20
98
9.8
81026
.75
10
43.6
01
06
4.3
21111
.38
115
5.8
011
79.
56
1201
.47
123
1.1
612
63.1
31
292
.47
1301
.14
1339
.42
13
76.7
313
93.8
314
06.2
81
42
3.7
714
55.1
01
48
6.0
81
525
.04
1543
.75
173
1.7
2
295
5.9
92
978
.61
307
8.5
531
36.
78
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
%T
600 800 1000 1000 1500 2000 3000
Wavenumbers (cm-1)
654.
77
758.
12
822.
80
88
0.2
9
927.
8795
5.22
989.
51
1064
.24
1154
.30
1177
.72
1204
.70
1230
.10
12
59.1
812
95.6
413
38.
40
13
55.0
913
90.6
51
407
.10
145
0.4
114
87.5
615
24.
87
15
43.7
6
172
7.1
6
2874
.78
2962
.77
30
24.7
830
51.
12
3079
.53
0
10
20
30
40
50
60
70
80
90
100
%T
600 800 1000 1000 1500 2000 3000
Wavenumbers (cm-1)
compound 3d
compound 3f
654
.01
758.
64
823
.7684
7.01
882.
97
943.
05
991
.05
104
4.6
9
1166
.70
1215
.65
1231
.9712
63.4
71
30
1.0
113
42.4
013
61.9
71382
.44
13
92.5
614
07.5
91
452
.38
14
85.
67
1526
.381
545
.52
172
7.4
9
293
0.5
429
76.
3930
56.4
630
80.
48
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
%T
600 800 1000 1000 1500 2000 3000
Wavenumbers (cm-1)
56
0.6
9
615.
64
67
6.79
74
2.9
37
57.
74
825.
7784
8.42
946.
5796
2.35
997.
0510
42.1
2
1145
.38
11
81.
51
1212
.32
1264
.33
1294
.17
132
9.0
2137
4.5
91
398
.39
1411
.89
144
1.5
614
65.5
914
95.
62
15
39.7
4
169
2.4
717
37.
07
296
2.0
93006
.52
3064
.55
31
04.
45
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
%T
600 800 1000 1000 1500 2000 3000
Wavenumbers (cm-1)
compound 3g
compound 3h
407
22
507.
17
556.
67
61
3.6
7
67
7.0
3
740.
8075
3.9
7
805
.19
82
5.6
58
59.
41
891
.63
945.
22
995.
7610
23.
95
10
42.9
310
56.5
6
1131
.53
114
2.2
61
201
.93
1257
.73
1292
.01
1322
.21
1350
.71
14
08.
09
144
5.5
81
465
.01
1477
.52
1500
.05
15
36.8
016
09.6
4
168
7.1
017
28.
91
2910
.36
294
1.4
829
90.5
2
31
42.0
1
0
10
20
30
40
50
60
70
80
90
100
%T
600 800 1000 1000 1500 2000 3000
Wavenumbers (cm-1)
510.
52
56
2.3
1
66
9.89
745
.73
757.
71
825
.85
858
.37
89
0.06
928.
8894
8.77
998.
6310
20.
13
10
62.
51
114
6.3
112
09.3
512
32.1
312
65.
16
12
92.4
913
25.3
813
58.7
31
393
.99
14
14.7
114
64.3
71
50
5.3
815
42.
51
172
8.4
2
287
5.6
72
936
.91
2962
.92
3034
.84
306
6.9
931
01.
52
-0
10
20
30
40
50
60
70
80
90
100
%T
600 800 1000 1000 1500 2000 3000
Wavenumbers (cm-1)
compound 3i
compound 3p
75
7.54
827.
75844
.4788
3.8
2
946.
87
998.
74
1142
.751
158
.71
1224
.90
1261
.80
1290
.08
1317
.65
1365
.90
1377
.06
1398
.11
1415
.95
1441
.40
14
55.7
014
63.5
914
98.6
11
537
.03
1633
.55
1644
.59
1651
.10
1660
.30
1682
.13
1698
.67
1727
.73
2978
.69
3109
.14
3436
.93
55
60
65
70
75
80
85
90
95
100
%T
800 1000 1000 1500 2000 3000
Wavenumbers (cm-1)
62
0.53
635
.36
71
2.6
37
38.
94
76
5.87
788
.0680
8.4
68
20.
21
829
.26
84
6.29
876
.99
889
.11
89
8.41
93
5.5
195
8.3
19
80.
10
10
52.7
810
62.
99
110
8.3
311
35.2
21
176
.23
12
05.8
012
51.
76
1288
.55
134
0.4
81
372
.90
138
5.0
31
416
.54
14
35.
52
1455
.53
146
9.3
514
94.
49
151
8.9
4
159
3.2
316
19.0
716
33.5
816
44.6
51
651
.21
16
67.
75
168
2.3
21
69
8.8
017
25.
46
285
1.4
329
23.
54
295
7.1
53
09
3.0
3
344
4.2
83
56
3.9
2
38
50.
65
64
66
68
70
72
74
76
78
80
82
84
86
88
90
92
94
96
98
100
%T
600 800 1000 1000 1500 2000 3000
Wavenumbers (cm-1)
compound 3q
compound 3r
42
7.1
159
1.0
56
21
.30
63
8.1
6
73
9.6
47
64
.14
79
1.9
98
23
.96
83
3.3
38
50
.58
86
0.2
08
89
.30
90
8.7
99
38
.16
98
3.3
01
01
3.3
11
05
1.3
31
06
0.7
61
09
3.1
41
11
4.6
41
13
3.9
51
16
7.1
81
19
9.1
21
25
8.5
01
29
4.5
31
33
7.4
31
38
3.2
51
39
5.6
01
41
2.6
91
44
5.4
21
46
6.8
31
51
0.1
1
15
92
.12
16
17
.71
17
15
.701
73
0.9
1
29
34
.54
29
76
.65
30
95
.45
31
21.
91
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
%T
600 800 1000 1000 1500 2000 3000
Wavenumbers (cm-1)
73
9.0
178
8.3
382
0.2
18
29
.35
84
6.3
48
77
.07
88
9.0
98
98
.52
935
.589
58
.42
980
.30
10
52
.82
10
63
.12
11
08
.27
11
35
.50
11
76.3
51
20
6.3
31
25
1.8
21
288
.44
13
40
.55
13
84
.45
14
16
.50
14
35
.60
14
55
.42
14
69
.361
49
4.5
21
518
.96
15
93
.32
16
19
.411
63
3.4
61
64
4.4
81
65
1.1
01
682
.01
16
98
.80
17
25
.20
29
23
.98
29
57
.55
30
93
.26
34
36
.24
35
84.5
93
59
2.4
63
625
.90
72
74
76
78
80
82
84
86
88
90
92
94
96
98
100
%T
600 800 1000 1000 1500 2000 3000
Wavenumbers (cm-1)
compound 3s
[1] J. Xu, C. Qian, B. Liu, Q. Wu and X. Lin, Tetrahedron, 2007, 63, 986‐990.
[2] Y. Cai, Q. Wu, Y. Xiao, D. Lv, X. Lin, J. Biotechnol., 2006, 121, 330‐337.
[3] J. Wen, Y. Luo, H. Zhang, H. Zhao, C. Zhou, G. Cai, Chin. Chem. Lett., 2016, 27, 391‐394.
74
0.7
47
63
.36
79
6.1
98
20
.54
84
3.8
1
88
4.4
790
8.9
693
6.4
8
98
6.0
2
10
61
.65
11
57
.07
12
30
.85
12
90
.18
13
45
.34
13
70
.70
13
88
.92
14
09
.89
14
43
.10
14
68
.941
49
3.9
115
16.
55
15
90
.53
16
17
.94
17
25
.05
29
77
.9630
66
.60
31
01
.65
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
%T
600 800 1000 1000 1500 2000 3000
Wavenumbers (cm-1)