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Furopyridinediones as novel inhibitors of -glucosidases
Rajanikant Mamidala,4 Chandramohan Bathula,1 Chiranjeevi Thulluri,2 Rahul Agarwal,3, Kunal Kumar Jha,1 Parthapratim Munshi,1 Subhabrata Sen1, *, Uma Adepally,2 Ashutosh Singh,3 M.
Thirumalacharya4
1 Department of Chemistry, School of Natural Sciences, Shiv Nadar University, Chithera, Dadri, UP 201314, India
2 Department of Life Science, School of Natural Sciences, Shiv Nadar University, Chithera, Dadri, UP 201314, India
3 Institute of Science and Technology, Jawaharlal Nehru Technological University Hyderabad, Kukatpally, Hyderabad500085, Telengana, India
4 Department of Chemistry, Jawaharlal Nehru Technological University Hyderabad, Kukatpally, Hyderabad500085, Telengana, India
1
Electronic Supplementary Material (ESI) for RSC Advances.This journal is © The Royal Society of Chemistry 2015
Contents
1. 1H-NMR spectra of compound 12a-v---------Page 1-22 2. 13C-NMR spectra of compound 12a-v----------Page 23-44
1
Pages 2-24Pages 25-45
2
Pages 3-24Pages 25-45
3. 12p a-glucosidase assay, LB plotAlpha-glucosidase assay, LB plot and COS cell assay 12p----Pages 46-48
2 3
3 4
4 5
5 6
6 7
7 8
8 9
9 10
10 11
11 12
12 13
ab
un
dan
ce
01
.02
.03
.04
.05
.06
.07
.08
.09
.01
0.0
11
.0
X : parts per Million : 1H
14.0 13.0 12.0 11.0 10.0 9.0 8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0
12.2
873
8
.3584
8
.3405
8
.0368
8
.0190
7
.1216
6
.8152
3
.3452
2
.5001
2
.3709
2
.3572
−
0.0
587
2.8
3
2.3
9
2
1.1
4
1.0
8
0.9
1
150115_PF_19_PROTON−6.jdf
Filename = 150115_PF_19_PROTON−6Author = deltaExperiment = single_pulse.ex2Sample_id = 150115_PF_19Solvent = DMSO−D6Creation_time = 15−JAN−2015 19:45:38Revision_time = 16−JAN−2015 10:44:10Current_time = 16−JAN−2015 10:44:37
Comment = 150115_PF_19Data_format = 1D COMPLEXDim_size = 13107Dim_title = 1HDim_units = [ppm]Dimensions = XSite = ECX 500Spectrometer = JNM−ECX500
Field_strength = 11.7473579[T] (500[MHX_acq_duration = 1.4548992[s]X_domain = 1HX_freq = 500.15991521[MHz]X_offset = 7.5[ppm]X_points = 16384X_prescans = 1X_resolution = 0.68733284[Hz]X_sweep = 11.26126126[kHz]Irr_domain = 1HIrr_freq = 500.15991521[MHz]Irr_offset = 5.0[ppm]Tri_domain = 1HTri_freq = 500.15991521[MHz]Tri_offset = 5.0[ppm]Clipped = FALSEMod_return = 1Scans = 16Total_scans = 16
X_90_width = 11.21[us]X_acq_time = 1.4548992[s]X_angle = 45[deg]X_atn = 3.4[dB]X_pulse = 5.605[us]Irr_mode = OffTri_mode = OffDante_presat = FALSEInitial_wait = 1[s]Recvr_gain = 48Relaxation_delay = 4[s]Repetition_time = 5.4548992[s]Temp_get = 18[dC]
−−−− PROCESSING PARAMETERS −−−−dc_balance : 0 : FALSEsexp : 0.2[Hz] : 0.0[s]fft : 1 : TRUE : TRUEmachinephaseppmphase : 4.25 : −14 : 73.28704[%]reference : 2.462[ppm] : 2.5[ppm]
Derived from: 150115_PF_19_PROTON−1.jdf
13 14
14 15
15 16
ab
un
dan
ce
01
.02
.03
.04
.05
.06
.07
.0
X : parts per Million : 1H
13.0 12.0 11.0 10.0 9.0 8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0
12.1
718
7
.8567
7
.4733
7
.3414
7
.1944
6
.7065
3
.3534
2
.5000
2
.4038
2
.3214
2
.2293
2.9
2.7
1.0
0.9
0.9
300615_PY_2_Thio_PROTON−4.jdf
Filename = 300615_PY_2_Thio_PROTAuthor = deltaExperiment = single_pulse.ex2Sample_id = 300615_PY_2_ThioSolvent = DMSO−D6Creation_time = 1−JUL−2015 09:50:53Revision_time = 1−JUL−2015 11:48:15Current_time = 1−JUL−2015 11:48:48
Data_format = 1D COMPLEXDim_size = 13107Dim_title = 1HDim_units = [ppm]Dimensions = XSite = ECX 500Spectrometer = JNM−ECX500
Field_strength = 11.7473579[T] (500[MHX_acq_duration = 1.4548992[s]X_domain = 1HX_freq = 500.15991521[MHz]X_offset = 7.5[ppm]X_points = 16384X_prescans = 1X_resolution = 0.68733284[Hz]X_sweep = 11.26126126[kHz]Irr_domain = 1HIrr_freq = 500.15991521[MHz]Irr_offset = 5.0[ppm]Tri_domain = 1HTri_freq = 500.15991521[MHz]Tri_offset = 5.0[ppm]Clipped = FALSEMod_return = 1Scans = 8Total_scans = 8
X_90_width = 11.6[us]X_acq_time = 1.4548992[s]X_angle = 45[deg]X_atn = 3.4[dB]X_pulse = 5.8[us]Irr_mode = OffTri_mode = OffDante_presat = FALSEInitial_wait = 1[s]Recvr_gain = 40Relaxation_delay = 4[s]Repetition_time = 5.4548992[s]Temp_get = 18.4[dC]
−−−− PROCESSING PARAMETERS −−−−dc_balance : 0 : FALSEsexp : 0.2[Hz] : 0.0[s]fft : 1 : TRUE : TRUEmachinephaseppm
Derived from: 300615_PY_2_Thio_PROTON−1.
16 17
10 9 8 7 6 5 4 3 2 1 ppm
0.8573
0.8977
0.9399
0.9595
0.9835
1.0104
1.0623
1.0955
1.2278
1.4516
1.4957
1.5105
1.5345
1.5716
1.6378
1.7612
1.8237
1.8507
1.9018
1.9079
1.9157
1.9613
2.3381
2.5093
2.8448
2.8921
2.9396
2.9628
3.0031
3.0340
3.2047
3.3572
3.6204
3.7015
3.8009
3.8213
3.8868
3.8983
3.9185
3.9466
4.0069
4.0250
4.0355
4.0618
4.1138
5.1521
6.6763
6.9801
6.9844
7.0006
7.2647
7.4276
7.6459
7.6779
7.7050
7.7467
7.7948
7.9454
0.3
50
3.0
45
0.9
50
2.1
72
0.9
07
NAME ShantanuEXPNO 2
PROCNO 1
Date_ 20150515
Time 11.41
INSTRUM SpectPROBHD 5 mm PABBO BB-
PULPROG zg30
TD 65536
SOLVENT DMSO
NS 16
DS 2SWH 10330.578 Hz
FIDRES 0.157632 Hz
AQ 3.1719923 sec
RG 144
DW 48.400 usec
DE 6.50 usecTE 299.1 K
D1 1.00000000 sec
TD0 1
======== CHANNEL f1 ========
NUC1 1HP1 10.90 usec
PL1 1.10 dB
PL1W 18.99148560 W
SFO1 500.1330885 MHz
SI 32768
SF 500.1300000 MHzWDW EM
SSB 0
LB 0.30 Hz
GB 0
PC 1.00
1 H NMR of PF-17
17 18
ab
un
dan
ce
01
.02
.03
.04
.05
.06
.07
.08
.09
.01
0.0
11
.01
2.0
13
.01
4.0
15
.01
6.0
17
.01
8.0
19
.02
0.0
21
.0
X : parts per Million : 1H
14.0 13.0 12.0 11.0 10.0 9.0 8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0
12.2
048
8
.1769
7
.8361
7
.7303
7
.7166
7
.4623
7
.4472
7
.4321
7
.3496
7
.3290
6
.7202
5
.1371
3
.3465
2
.5027
2
.5000
2
.3365
3
2.1
3
2.1
22.0
9
1.1
7
1.0
6
1.0
4
1.0
2
160115_PF_22−4.jdf
Filename = 160115_PF_22−4.jdfAuthor = deltaExperiment = single_pulse.ex2Sample_id = 160115_PF_22Solvent = DMSO−D6Creation_time = 16−JAN−2015 17:38:10Revision_time = 17−JAN−2015 09:38:05Current_time = 17−JAN−2015 09:38:29
Data_format = 1D COMPLEXDim_size = 13107Dim_title = 1HDim_units = [ppm]Dimensions = XSite = ECX 500Spectrometer = JNM−ECX500
Field_strength = 11.7473579[T] (500[MHX_acq_duration = 1.4548992[s]X_domain = 1HX_freq = 500.15991521[MHz]X_offset = 7.5[ppm]X_points = 16384X_prescans = 1X_resolution = 0.68733284[Hz]X_sweep = 11.26126126[kHz]Irr_domain = 1HIrr_freq = 500.15991521[MHz]Irr_offset = 5.0[ppm]Tri_domain = 1HTri_freq = 500.15991521[MHz]Tri_offset = 5.0[ppm]Clipped = FALSEMod_return = 1Scans = 16Total_scans = 16
X_90_width = 11.21[us]X_acq_time = 1.4548992[s]X_angle = 45[deg]X_atn = 3.4[dB]X_pulse = 5.605[us]Irr_mode = OffTri_mode = OffDante_presat = FALSEInitial_wait = 1[s]Recvr_gain = 40Relaxation_delay = 5[s]Repetition_time = 6.4548992[s]Temp_get = 19.5[dC]
−−−− PROCESSING PARAMETERS −−−−dc_balance : 0 : FALSEsexp : 0.2[Hz] : 0.0[s]trapezoid3 : 0[%] : 80[%] : 100[%]zerofill : 1fft : 1 : TRUE : TRUEmachinephaseppm
Derived from: 160115_PF_22−1.jdf
18 19
abundance
0 1.0 2.0 3.0
X : p
arts p
er Millio
n : 1
H
12.0
11.0
10.0
9.0
8.0
7.0
6.0
5.0
4.0
3.0
2.0
1.0
0−
1.0
−2.0
12.1644
8.9543
8.5089 8.4998 8.2983 8.1105 8.0911 7.8701
7.4591 7.4477 7.4396 7.4270 7.3057 6.7195
3.2816
2.4699 2.4653 2.4619 2.3142
0.1116 −0.0350 −0.1861
RG
09−
3.jd
f
19 20
ab
un
dan
ce
00
.10
.20
.30
.40
.50
.60
.70
.80
.91
.01
.11
.21
.31
.4
X : parts per Million : 1H
13.0 12.0 11.0 10.0 9.0 8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0
12.1
746
8
.5658
8
.1618
8
.0725
7
.8196
7
.3263
6
.9030
6
.7450
3
.8921
3
.3342
2
.5000
2
.3433
3.1
2.9
2.2
2.0
1.1
1.11
0.9
010715_PF_11−4.jdf
Filename = 010715_PF_11−4.jdfAuthor = deltaExperiment = single_pulse.ex2Sample_id = 010715_PF_11Solvent = DMSO−D6Creation_time = 1−JUL−2015 15:58:42Revision_time = 1−JUL−2015 15:19:05Current_time = 1−JUL−2015 15:19:16
Data_format = 1D COMPLEXDim_size = 13107Dim_title = 1HDim_units = [ppm]Dimensions = XSite = ECX 500Spectrometer = JNM−ECX500
Field_strength = 11.7473579[T] (500[MHX_acq_duration = 1.4548992[s]X_domain = 1HX_freq = 500.15991521[MHz]X_offset = 7.5[ppm]X_points = 16384X_prescans = 1X_resolution = 0.68733284[Hz]X_sweep = 11.26126126[kHz]Irr_domain = 1HIrr_freq = 500.15991521[MHz]Irr_offset = 5.0[ppm]Tri_domain = 1HTri_freq = 500.15991521[MHz]Tri_offset = 5.0[ppm]Clipped = FALSEMod_return = 1Scans = 8Total_scans = 8
X_90_width = 11.6[us]X_acq_time = 1.4548992[s]X_angle = 45[deg]X_atn = 3.4[dB]X_pulse = 5.8[us]Irr_mode = OffTri_mode = OffDante_presat = FALSEInitial_wait = 1[s]Recvr_gain = 46Relaxation_delay = 5[s]Repetition_time = 6.4548992[s]Temp_get = 22.1[dC]
−−−− PROCESSING PARAMETERS −−−−dc_balance : 0 : FALSEsexp : 0.2[Hz] : 0.0[s]trapezoid3 : 0[%] : 80[%] : 100[%]zerofill : 1fft : 1 : TRUE : TRUEmachinephaseppm
Derived from: 010715_PF_11−1.jdf
20 21
ab
un
dan
ce
01
.02
.03
.0
X : parts per Million : 1H
13.0 12.0 11.0 10.0 9.0 8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0
11.9
684
8
.4270
8
.1632
8
.1536
7
.9048
7
.3070
7
.1449
7
.0033
6
.9937
6
.7161
3
.5691
3
.4400
3
.3438
2
.5000
2
.3406
2
.2911
2
.0836
1
.4295
9.0
5
3.8
83.5
3
3.3
4
1.0
2
1.0
1
0.9
9
0.9
70.9
6
0.9
6
0.5
7
150115_PF_21_PROTON−4.jdf
Filename = 150115_PF_21_PROTON−4Author = deltaExperiment = single_pulse.ex2Sample_id = 150115_PF_21Solvent = DMSO−D6Creation_time = 15−JAN−2015 17:48:37Revision_time = 16−JAN−2015 10:22:34Current_time = 16−JAN−2015 10:23:11
Comment = 150115_PF_21Data_format = 1D COMPLEXDim_size = 13107Dim_title = 1HDim_units = [ppm]Dimensions = XSite = ECX 500Spectrometer = JNM−ECX500
Field_strength = 11.7473579[T] (500[MHX_acq_duration = 1.4548992[s]X_domain = 1HX_freq = 500.15991521[MHz]X_offset = 7.5[ppm]X_points = 16384X_prescans = 1X_resolution = 0.68733284[Hz]X_sweep = 11.26126126[kHz]Irr_domain = 1HIrr_freq = 500.15991521[MHz]Irr_offset = 5.0[ppm]Tri_domain = 1HTri_freq = 500.15991521[MHz]Tri_offset = 5.0[ppm]Clipped = FALSEMod_return = 1Scans = 16Total_scans = 16
X_90_width = 11.21[us]X_acq_time = 1.4548992[s]X_angle = 45[deg]X_atn = 3.4[dB]X_pulse = 5.605[us]Irr_mode = OffTri_mode = OffDante_presat = FALSEInitial_wait = 1[s]Recvr_gain = 44Relaxation_delay = 4[s]Repetition_time = 5.4548992[s]Temp_get = 18.2[dC]
−−−− PROCESSING PARAMETERS −−−−dc_balance : 0 : FALSEsexp : 0.2[Hz] : 0.0[s]fft : 1 : TRUE : TRUEmachinephaseppm
Derived from: 150115_PF_21_PROTON−1.jdf
21 22
ab
un
dan
ce
01
.02
.03
.04
.05
.06
.07
.08
.09
.01
0.0
11
.01
2.0
13
.01
4.0
X : parts per Million : 1H
13.0 12.0 11.0 10.0 9.0 8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0
1
2.2
103
7
.87
05
7
.72
48
7
.63
96
7
.62
45
7
.61
21
7
.29
88
7
.23
70
7
.23
01
7
.21
22
7
.20
54
7
.13
66
7
.10
37
6
.74
22
3
.34
38
2
.50
00
2
.33
78
2
.08
22
3.1
2
2.6
3
2.0
1.0
7
1.0
310.8
5
0.8
30.8
3
150115_PF_20_PROTON−4.jdf
Filename = 150115_PF_20_PROTON−4Author = deltaExperiment = single_pulse.ex2Sample_id = 150115_PF_20Solvent = DMSO−D6Creation_time = 15−JAN−2015 21:41:25Revision_time = 16−JAN−2015 10:53:34Current_time = 16−JAN−2015 10:54:00
Comment = 150115_PF_20Data_format = 1D COMPLEXDim_size = 13107Dim_title = 1HDim_units = [ppm]Dimensions = XSite = ECX 500Spectrometer = JNM−ECX500
Field_strength = 11.7473579[T] (500[MHX_acq_duration = 1.4548992[s]X_domain = 1HX_freq = 500.15991521[MHz]X_offset = 7.5[ppm]X_points = 16384X_prescans = 1X_resolution = 0.68733284[Hz]X_sweep = 11.26126126[kHz]Irr_domain = 1HIrr_freq = 500.15991521[MHz]Irr_offset = 5.0[ppm]Tri_domain = 1HTri_freq = 500.15991521[MHz]Tri_offset = 5.0[ppm]Clipped = FALSEMod_return = 1Scans = 16Total_scans = 16
X_90_width = 11.21[us]X_acq_time = 1.4548992[s]X_angle = 45[deg]X_atn = 3.4[dB]X_pulse = 5.605[us]Irr_mode = OffTri_mode = OffDante_presat = FALSEInitial_wait = 1[s]Recvr_gain = 46Relaxation_delay = 4[s]Repetition_time = 5.4548992[s]Temp_get = 18.3[dC]
−−−− PROCESSING PARAMETERS −−−−dc_balance : 0 : FALSEsexp : 0.2[Hz] : 0.0[s]fft : 1 : TRUE : TRUEmachinephaseppm
Derived from: 150115_PF_20_PROTON−1.jdf
22 23
NAME P165CB-Z00763-045-AEXPNO 1PROCNO 1Date_ 20140328Time 16.48INSTRUM spectPROBHD 5 mm DUL 13C-1PULPROG zg30TD 32768SOLVENT DMSONS 8DS 0SWH 8223.685 HzFIDRES 0.250967 HzAQ 1.9923444 secRG 128DW 60.800 usecDE 6.50 usecTE 300.0 KD1 1.00000000 secTD0 1
======== CHANNEL f1 ========NUC1 1HP1 9.90 usecPL1 -2.80 dBPL1W 20.58109283 WSFO1 400.2624718 MHzSI 65536SF 400.2600044 MHzWDW EMSSB 0LB 0.30 HzGB 0PC 1.00
P165GB-Z00763-045-A DMSO P.S
14 13 12 11 10 9 8 7 6 5 4 3 2 1 0 ppm
1.679
2.145
2.500
3.328
4.054
6.614
7.251
7.261
7.888
7.901
7.911
7.919
10.697
6.17
3.00
3.89
0.92
0.99
1.94
0.96
23 24
0102030405060708090100110120130140150160170180f1 (ppm)
-5E+06
0
5E+06
1E+07
2E+07
2E+07
2E+07
3E+07
4E+07
4E+07
4E+07
5E+07
6E+07
6E+07
6E+078.9.15 13C NMR of RM-1
19.7
219
.92
21.1
5
97.7
9
106.
01
109.
28
130.
5213
0.95
131.
3813
5.39
135.
61
143.
16
155.
6715
6.65
157.
98
164.
84
24 25
0102030405060708090100110120130140150160170180f1 (ppm)
-5E+06
0
5E+06
1E+07
2E+07
2E+07
2E+07
3E+07
4E+07
4E+07
4E+07
5E+07
6E+07
6E+07
6E+07
7E+07
8E+07
8E+07
8E+078.9.15 13C NMR of RM-2
12.3
216
.39
20.3
8
56.0
4
97.4
2
105.
4510
8.88
110.
47
123.
9612
4.98
129.
86
138.
47
141.
76
155.
1115
6.71
158.
29
164.
90
25 26
0102030405060708090100110120130140150160170180190f1 (ppm)
-2E+07
0
2E+07
4E+07
6E+07
8E+07
1E+08
1E+08
1E+08
2E+08
2E+08
2E+08
2E+08
2E+08
3E+08
3E+08
3E+08
3E+088.9.15 13C NMR of RM-5
19.9
6
97.5
2
105.
2710
6.59
126.
6412
7.31
127.
5612
9.69
130.
6413
2.57
133.
29
145.
45
156.
0915
7.02
157.
98
164.
19
26 27
-100102030405060708090100110120130140150160170180190200210f1 (ppm)
0
5E+07
1E+08
2E+08
2E+08
2E+08
3E+08
4E+08
4E+08
8.9.15 13C NMR of RM-6
19.6
520
.00
68.2
8
97.0
399
.84
105.
1410
9.28
113.
3911
6.45
117.
66
124.
4012
4.67
140.
5814
6.05
148.
29
155.
0515
6.71
158.
35
164.
9016
7.43
168.
75
27 28
0102030405060708090100110120130140150160170f1 (ppm)
-1E+07
0
1E+07
2E+07
3E+07
4E+07
5E+07
6E+07
7E+07
8E+07
9E+07
1E+08
1E+08
1E+08
1E+08
1E+08
2E+08
2E+088.9.15 13C NMR of RM-20
19.0
120
.10
56.5
0
97.3
5
106.
10
109.
86
122.
61
129.
7113
1.52
132.
2313
2.74
135.
53
144.
10
156.
2715
6.54
158.
04
164.
30
28 29
0102030405060708090100110120130140150160170180190f1 (ppm)
-5E+07
0
5E+07
1E+08
2E+08
2E+08
2E+08
3E+08
4E+08
4E+08
4E+08
5E+08
6E+088.9.15 13C NMR of RM-21
20.0
9
97.3
4
106.
02
110.
45
123.
18
132.
4413
2.49
132.
58
143.
67
156.
1115
6.61
158.
10
164.
37
29 30
0102030405060708090100110120130140150160170f1 (ppm)
0
1E+07
2E+07
3E+07
4E+07
5E+07
6E+07
7E+07
8E+07
9E+07
8.9.15 13C NMR of RM-22
20.1
7
97.9
3
105.
9410
7.50
124.
3212
6.17
126.
8712
7.42
129.
1112
9.77
130.
2413
0.29
131.
7713
3.84
144.
54
155.
9215
6.75
158.
31
164.
75
30 31
0102030405060708090100110120130140150160170180f1 (ppm)
0
1E+07
2E+07
3E+07
4E+07
5E+07
6E+07
7E+07
8E+07
9E+07
1E+088.9.15 13C NMR of RM-18
20.0
7
97.2
3
106.
00
109.
32
118.
6211
8.76
118.
8511
8.99
128.
1113
0.88
143.
69
156.
2615
6.47
158.
00
164.
21
31 32
0102030405060708090100110120130140150160170180f1 (ppm)
-1E+07
0
1E+07
2E+07
3E+07
4E+07
5E+07
6E+07
7E+07
8E+07
9E+07
1E+08
1E+08
1E+08
1E+08
8.9.15 13C NMR of RM-16
20.0
2
97.0
5
102.
1810
5.50
109.
3910
9.73
112.
13
126.
6412
7.38
141.
64
148.
3314
8.90
155.
5015
6.64
158.
19
164.
55
32 33
102030405060708090100110120130140150160170180f1 (ppm)
0
5E+07
1E+08
2E+08
2E+08
2E+08
3E+08
4E+08
4E+08
4E+088.9.15 13C NMR of RM-27
20.0
7
97.3
4
105.
95
111.
82
129.
4912
9.83
130.
8613
3.21
143.
19
155.
9415
6.78
158.
17
164.
59
33 34
abun
danc
e
00.
10.
20.
3
X : parts per Million : 13C
210.0 200.0 190.0 180.0 170.0 160.0 150.0 140.0 130.0 120.0 110.0 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0 0
175
.907
5
166
.607
7
146
.853
8 1
43.0
862
130
.953
4 1
28.0
252
124
.152
6 1
19.9
557
54.
9521
43.
9353
39.
9960
39.
8338
39.
6622
39.
5000
39.
3378
39.
1662
39.
0040
22.
5504
160115_PF_19_13C_CARBON−4.jdf
Filename = 160115_PF_19_13C_CARBAuthor = deltaExperiment = single_pulse_decSample_id = 160115_PF_19_13CSolvent = DMSO−D6Creation_time = 17−JAN−2015 03:17:36Revision_time = 17−JAN−2015 09:53:20Current_time = 17−JAN−2015 09:53:43
Comment = 160115_PF_19_13CData_format = 1D COMPLEXDim_size = 26214Dim_title = 13CDim_units = [ppm]Dimensions = XSite = ECX 500Spectrometer = JNM−ECX500
Field_strength = 11.7473579[T] (500[MHX_acq_duration = 0.83361792[s]X_domain = 13CX_freq = 125.76529768[MHz]X_offset = 100[ppm]X_points = 32768X_prescans = 4X_resolution = 1.19959034[Hz]X_sweep = 39.3081761[kHz]Irr_domain = 1HIrr_freq = 500.15991521[MHz]Irr_offset = 5.0[ppm]Clipped = FALSEMod_return = 1Scans = 1024Total_scans = 1024
X_90_width = 8.62[us]X_acq_time = 0.83361792[s]X_angle = 30[deg]X_atn = 6.4[dB]X_pulse = 2.87333333[us]Irr_atn_dec = 21.68[dB]Irr_atn_noe = 21.68[dB]Irr_noise = WALTZDecoupling = TRUEInitial_wait = 1[s]Noe = TRUENoe_time = 2[s]Recvr_gain = 60Relaxation_delay = 2[s]Repetition_time = 2.83361792[s]Temp_get = 18.1[dC]
−−−− PROCESSING PARAMETERS −−−−dc_balance : 0 : FALSEsexp : 2.0[Hz] : 0.0[s]fft : 1 : TRUE : TRUEmachinephaseppm
Derived from: 160115_PF_19_13C_CARBON−1.
34 35
-100102030405060708090100110120130140150160170180190200210f1 (ppm)
-1E+07
0
1E+07
2E+07
3E+07
4E+07
5E+07
6E+07
7E+07
8E+07
9E+07
1E+08
1E+08
1E+08
1E+08
1E+08
2E+08
2E+08
2E+088.9.15 13C NMR of RM-7
19.6
519
.99
68.2
9
96.9
499
.84
104.
8010
6.85
109.
2910
9.59
112.
72
119.
2612
1.01
123.
0212
6.96
130.
6913
6.44
139.
38
154.
3515
5.05
155.
7915
8.38
158.
5616
4.88
167.
4116
8.77
35 36
36 37
abun
danc
e
00.
10.
20.
30.
40.
50.
60.
7
X : parts per Million : 13C
210.0 200.0 190.0 180.0 170.0 160.0 150.0 140.0 130.0 120.0 110.0 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0 0
166
.598
1 1
63.6
794
157
.746
6 1
55.2
952
140
.577
6
135
.627
2 1
32.0
980
132
.021
7 1
28.0
919
106
.010
7 1
05.5
624
96.
6822
39.
9960
39.
8338
39.
6622
39.
5000
39.
3283
39.
1662
38.
9945
19.
6412
300615_PY_2_Thio_CARBON−4.jdf
Filename = 300615_PY_2_Thio_CARBAuthor = deltaExperiment = single_pulse_decSample_id = 300615_PY_2_ThioSolvent = DMSO−D6Creation_time = 1−JUL−2015 10:39:45Revision_time = 1−JUL−2015 11:51:05Current_time = 1−JUL−2015 11:51:28
Data_format = 1D COMPLEXDim_size = 26214Dim_title = 13CDim_units = [ppm]Dimensions = XSite = ECX 500Spectrometer = JNM−ECX500
Field_strength = 11.7473579[T] (500[MHX_acq_duration = 0.83361792[s]X_domain = 13CX_freq = 125.76529768[MHz]X_offset = 100[ppm]X_points = 32768X_prescans = 4X_resolution = 1.19959034[Hz]X_sweep = 39.3081761[kHz]Irr_domain = 1HIrr_freq = 500.15991521[MHz]Irr_offset = 5.0[ppm]Clipped = FALSEMod_return = 1Scans = 1024Total_scans = 1024
X_90_width = 8.62[us]X_acq_time = 0.83361792[s]X_angle = 30[deg]X_atn = 6.4[dB]X_pulse = 2.87333333[us]Irr_atn_dec = 21.68[dB]Irr_atn_noe = 21.68[dB]Irr_noise = WALTZDecoupling = TRUEInitial_wait = 1[s]Noe = TRUENoe_time = 2[s]Recvr_gain = 60Relaxation_delay = 2[s]Repetition_time = 2.83361792[s]Temp_get = 19[dC]
−−−− PROCESSING PARAMETERS −−−−dc_balance : 0 : FALSEsexp : 2.0[Hz] : 0.0[s]fft : 1 : TRUE : TRUEmachinephaseppm
Derived from: 300615_PY_2_Thio_CARBON−1.
37 38
180 160 140 120 100 80 60 40 20 ppm
20.10
39.47
39.64
39.81
39.97
40.14
40.31
40.47
97.23
104.66
106.26
110.43
129.32
133.12
137.66
142.34
155.54
156.27
158.06
163.79
NAME ShantanuEXPNO 3PROCNO 1Date_ 20150515Time 13.31INSTRUM SpectPROBHD 5 mm PABBO BB-PULPROG zgpg30TD 65536SOLVENT DMSONS 2048DS 4SWH 29761.904 HzFIDRES 0.454131 HzAQ 1.1010548 secRG 144DW 16.800 usecDE 6.50 usecTE 299.9 KD1 2.00000000 secD11 0.03000000 secTD0 1
======== CHANNEL f1 ========NUC1 13CP1 8.75 usecPL1 -2.20 dBPL1W 116.72342682 WSFO1 125.7703643 MHz
======== CHANNEL f2 ========CPDPRG2 waltz16NUC2 1HPCPD2 80.00 usecPL2 1.10 dBPL12 18.50 dBPL13 18.50 dBPL2W 18.99148560 WPL12W 0.34558824 WPL13W 0.34558824 WSFO2 500.1320005 MHzSI 32768SF 125.7577890 MHzWDW EMSSB 0LB 1.00 HzGB 0PC 1.40
13C NMR of PF-17
38 39
abun
danc
e
00.
10.
20.
30.
40.
50.
60.
70.
80.
91.
01.
11.
21.
31.
41.
51.
61.
7
X : parts per Million : 13C
210.0200.0 190.0 180.0 170.0 160.0 150.0 140.0 130.0 120.0 110.0 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0 0
163
.584
0
157
.708
4 1
55.4
860
155
.190
3
141
.750
8 1
41.0
831
136
.476
1 1
34.2
823
130
.037
7 1
29.0
553
127
.615
0 1
27.0
236
126
.031
6
105
.781
8 1
05.6
292
99.
3720
96.
7204
39.
9960
39.
8338
39.
6622
39.
5000
39.
3283
39.
1662
39.
0040
19.
6317
160115_PF_22_13C−4.jdf
Filename = 160115_PF_22_13C−4.jdAuthor = deltaExperiment = single_pulse_decSample_id = 160115_PF_22_13CSolvent = DMSO−D6Creation_time = 16−JAN−2015 18:27:09Revision_time = 17−JAN−2015 09:35:15Current_time = 17−JAN−2015 09:35:35
Data_format = 1D COMPLEXDim_size = 26214Dim_title = 13CDim_units = [ppm]Dimensions = XSite = ECX 500Spectrometer = JNM−ECX500
Field_strength = 11.7473579[T] (500[MHX_acq_duration = 0.83361792[s]X_domain = 13CX_freq = 125.76529768[MHz]X_offset = 100[ppm]X_points = 32768X_prescans = 4X_resolution = 1.19959034[Hz]X_sweep = 39.3081761[kHz]Irr_domain = 1HIrr_freq = 500.15991521[MHz]Irr_offset = 5.0[ppm]Clipped = FALSEMod_return = 1Scans = 1024Total_scans = 1024
X_90_width = 8.62[us]X_acq_time = 0.83361792[s]X_angle = 30[deg]X_atn = 6.4[dB]X_pulse = 2.87333333[us]Irr_atn_dec = 21.68[dB]Irr_atn_noe = 21.68[dB]Irr_noise = WALTZDecoupling = TRUEInitial_wait = 1[s]Noe = TRUENoe_time = 2[s]Recvr_gain = 60Relaxation_delay = 2[s]Repetition_time = 2.83361792[s]Temp_get = 19.3[dC]
−−−− PROCESSING PARAMETERS −−−−dc_balance : 0 : FALSEsexp : 2.0[Hz] : 0.0[s]trapezoid3 : 0[%] : 80[%] : 100[%]zerofill : 1fft : 1 : TRUE : TRUEmachinephaseppm
Derived from: 160115_PF_22_13C−1.jdf
39 40
abun
danc
e
00.
10.
2
X : parts per Million : 13C
200.0 190.0 180.0 170.0 160.0 150.0 140.0 130.0 120.0 110.0 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0 0
169
.116
2
163
.526
8
157
.689
3 1
55.6
004
155
.161
7
148
.542
1 1
47.1
304
141
.321
6 1
38.4
982
133
.376
1 1
30.0
663
129
.770
7 1
28.1
110
123
.971
4
105
.638
7 1
05.5
243
96.
7585
93.
1054
40.
0055
39.
8338
39.
6717
39.
5000
39.
3378
39.
1662
39.
0040
19.
6508
160115_PF_9_13C_CARBON−4.jdf
Filename = 160115_PF_9_13C_CARBOAuthor = deltaExperiment = single_pulse_decSample_id = 160115_PF_9_13CSolvent = DMSO−D6Creation_time = 16−JAN−2015 20:12:19Revision_time = 17−JAN−2015 09:41:27Current_time = 17−JAN−2015 09:41:56
Comment = 160115_PF_9_13CData_format = 1D COMPLEXDim_size = 26214Dim_title = 13CDim_units = [ppm]Dimensions = XSite = ECX 500Spectrometer = JNM−ECX500
Field_strength = 11.7473579[T] (500[MHX_acq_duration = 0.83361792[s]X_domain = 13CX_freq = 125.76529768[MHz]X_offset = 100[ppm]X_points = 32768X_prescans = 4X_resolution = 1.19959034[Hz]X_sweep = 39.3081761[kHz]Irr_domain = 1HIrr_freq = 500.15991521[MHz]Irr_offset = 5.0[ppm]Clipped = FALSEMod_return = 1Scans = 2048Total_scans = 2048
X_90_width = 8.62[us]X_acq_time = 0.83361792[s]X_angle = 30[deg]X_atn = 6.4[dB]X_pulse = 2.87333333[us]Irr_atn_dec = 21.68[dB]Irr_atn_noe = 21.68[dB]Irr_noise = WALTZDecoupling = TRUEInitial_wait = 1[s]Noe = TRUENoe_time = 2[s]Recvr_gain = 60Relaxation_delay = 2[s]Repetition_time = 2.83361792[s]Temp_get = 19.2[dC]
−−−− PROCESSING PARAMETERS −−−−dc_balance : 0 : FALSEsexp : 2.0[Hz] : 0.0[s]fft : 1 : TRUE : TRUEmachinephaseppm
Derived from: 160115_PF_9_13C_CARBON−1.j
40 41
abun
danc
e
00.
10.
20.
30.
40.
50.
60.
70.
80.
91.
01.
1
X : parts per Million : 13C
180.0 170.0 160.0 150.0 140.0 130.0 120.0 110.0 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0 0
163
.545
9 1
62.8
877
157
.679
8 1
55.4
764
155
.152
1
144
.183
1 1
41.1
404
138
.412
4 1
36.9
626
136
.638
3 1
36.3
998
129
.608
5 1
26.4
227
124
.028
6
110
.694
0
105
.657
8
96.
7490
53.
2924
40.
0055
39.
8338
39.
6717
39.
5000
39.
3378
39.
1662
39.
0040
19.
6317
020715_PF_11_13C−4.jdf
Filename = 020715_PF_11_13C−4.jdAuthor = deltaExperiment = single_pulse_decSample_id = 020715_PF_11_13CSolvent = DMSO−D6Creation_time = 2−JUL−2015 15:56:47Revision_time = 2−JUL−2015 15:18:03Current_time = 2−JUL−2015 15:18:18
Data_format = 1D COMPLEXDim_size = 26214Dim_title = 13CDim_units = [ppm]Dimensions = XSite = ECX 500Spectrometer = JNM−ECX500
Field_strength = 11.7473579[T] (500[MHX_acq_duration = 0.83361792[s]X_domain = 13CX_freq = 125.76529768[MHz]X_offset = 100[ppm]X_points = 32768X_prescans = 4X_resolution = 1.19959034[Hz]X_sweep = 39.3081761[kHz]Irr_domain = 1HIrr_freq = 500.15991521[MHz]Irr_offset = 5.0[ppm]Clipped = FALSEMod_return = 1Scans = 442Total_scans = 442
X_90_width = 8.62[us]X_acq_time = 0.83361792[s]X_angle = 90[deg]X_atn = 6.4[dB]X_pulse = 8.62[us]Irr_atn_dec = 21.68[dB]Irr_atn_noe = 21.68[dB]Irr_noise = WALTZDecoupling = TRUEInitial_wait = 1[s]Noe = TRUENoe_time = 2[s]Recvr_gain = 60Relaxation_delay = 2[s]Repetition_time = 2.83361792[s]Temp_get = 20.7[dC]
−−−− PROCESSING PARAMETERS −−−−dc_balance : 0 : FALSEsexp : 2.0[Hz] : 0.0[s]trapezoid3 : 0[%] : 80[%] : 100[%]zerofill : 1fft : 1 : TRUE : TRUEmachinephaseppm
Derived from: 020715_PF_11_13C−1.jdf
41 42
abun
danc
e
00.
10.
20.
30.
40.
50.
60.
70.
8
X : parts per Million : 13C
210.0 200.0 190.0 180.0 170.0 160.0 150.0 140.0 130.0 120.0 110.0 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0 0
163
.498
2 1
59.5
684
157
.679
8 1
55.5
909
155
.114
0 1
53.9
789
148
.408
5 1
42.6
569
141
.264
3 1
40.0
244
136
.523
8 1
30.0
282
129
.484
5
110
.674
9
105
.686
4 1
05.5
338
103
.673
8 9
6.72
04
79.
0363
44.
4885
39.
9960
39.
8338
39.
6622
39.
5000
39.
3283
39.
1662
38.
9945
28.
0826
19.
6603
150115_PF_21_13C_CARBON−4.jdf
Filename = 150115_PF_21_13C_CARBAuthor = deltaExperiment = single_pulse_decSample_id = 150115_PF_21Solvent = DMSO−D6Creation_time = 15−JAN−2015 18:37:35Revision_time = 16−JAN−2015 10:24:55Current_time = 16−JAN−2015 10:28:57
Comment = 150115_PF_21_13CData_format = 1D COMPLEXDim_size = 26214Dim_title = 13CDim_units = [ppm]Dimensions = XSite = ECX 500Spectrometer = JNM−ECX500
Field_strength = 11.7473579[T] (500[MHX_acq_duration = 0.83361792[s]X_domain = 13CX_freq = 125.76529768[MHz]X_offset = 100[ppm]X_points = 32768X_prescans = 4X_resolution = 1.19959034[Hz]X_sweep = 39.3081761[kHz]Irr_domain = 1HIrr_freq = 500.15991521[MHz]Irr_offset = 5.0[ppm]Clipped = FALSEMod_return = 1Scans = 1024Total_scans = 1024
X_90_width = 8.62[us]X_acq_time = 0.83361792[s]X_angle = 30[deg]X_atn = 6.4[dB]X_pulse = 2.87333333[us]Irr_atn_dec = 21.68[dB]Irr_atn_noe = 21.68[dB]Irr_noise = WALTZDecoupling = TRUEInitial_wait = 1[s]Noe = TRUENoe_time = 2[s]Recvr_gain = 60Relaxation_delay = 2[s]Repetition_time = 2.83361792[s]Temp_get = 18.1[dC]
−−−− PROCESSING PARAMETERS −−−−dc_balance : 0 : FALSEsexp : 2.0[Hz] : 0.0[s]fft : 1 : TRUE : TRUEmachinephaseppm
Derived from: 150115_PF_21_13C_CARBON−1.
42 43
abun
danc
e
00.
10.
20.
30.
40.
50.
60.
70.
80.
91.
0
X : parts per Million : 13C
200.0 190.0 180.0 170.0 160.0 150.0 140.0 130.0 120.0 110.0 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0 0
163
.584
0 1
60.6
844
157
.708
4 1
55.4
478
155
.209
4
141
.178
5 1
40.1
674
136
.228
1 1
33.5
192
129
.236
5 1
28.6
928
128
.196
8 1
27.9
393
122
.216
3 1
15.7
112
115
.539
5
105
.705
5 1
05.5
815
96.
7776
39.
9960
39.
8338
39.
6622
39.
5000
39.
3283
39.
1662
38.
9945
30.
7152
19.
6412
150115_PF_20_13C_CARBON−4.jdf
Filename = 150115_PF_20_13C_CARBAuthor = deltaExperiment = single_pulse_decSample_id = 150115_PF_20Solvent = DMSO−D6Creation_time = 15−JAN−2015 22:30:17Revision_time = 16−JAN−2015 10:48:54Current_time = 16−JAN−2015 10:49:19
Comment = 150115_PF_20_13CData_format = 1D COMPLEXDim_size = 26214Dim_title = 13CDim_units = [ppm]Dimensions = XSite = ECX 500Spectrometer = JNM−ECX500
Field_strength = 11.7473579[T] (500[MHX_acq_duration = 0.83361792[s]X_domain = 13CX_freq = 125.76529768[MHz]X_offset = 100[ppm]X_points = 32768X_prescans = 4X_resolution = 1.19959034[Hz]X_sweep = 39.3081761[kHz]Irr_domain = 1HIrr_freq = 500.15991521[MHz]Irr_offset = 5.0[ppm]Clipped = FALSEMod_return = 1Scans = 1024Total_scans = 1024
X_90_width = 8.62[us]X_acq_time = 0.83361792[s]X_angle = 30[deg]X_atn = 6.4[dB]X_pulse = 2.87333333[us]Irr_atn_dec = 21.68[dB]Irr_atn_noe = 21.68[dB]Irr_noise = WALTZDecoupling = TRUEInitial_wait = 1[s]Noe = TRUENoe_time = 2[s]Recvr_gain = 60Relaxation_delay = 2[s]Repetition_time = 2.83361792[s]Temp_get = 18.4[dC]
−−−− PROCESSING PARAMETERS −−−−dc_balance : 0 : FALSEsexp : 2.0[Hz] : 0.0[s]fft : 1 : TRUE : TRUEmachinephaseppm
Derived from: 150115_PF_20_13C_CARBON−1.
43 44
abun
danc
e
00.
10.
20.
3
X : parts per Million : 13C
200.0 190.0 180.0 170.0 160.0 150.0 140.0 130.0 120.0 110.0 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0 0
167
.313
5
159
.110
5 1
58.8
244
146
.062
1 1
43.5
726
139
.642
8
132
.098
0 1
31.6
116
130
.381
1 1
28.1
873
95.
6521
48.
5042
40.
0055
39.
8338
39.
6717
39.
5000
39.
3378
39.
1662
39.
0040
25.
3642
23.
5138
18.
9926
160115_PF_5_13C_CARBON−4.jdf
Filename = 160115_PF_5_13C_CARBOAuthor = deltaExperiment = single_pulse_decSample_id = 160115_PF_5_13CSolvent = DMSO−D6Creation_time = 16−JAN−2015 23:41:22Revision_time = 17−JAN−2015 09:44:32Current_time = 17−JAN−2015 09:44:58
Comment = 160115_PF_5_13CData_format = 1D COMPLEXDim_size = 26214Dim_title = 13CDim_units = [ppm]Dimensions = XSite = ECX 500Spectrometer = JNM−ECX500
Field_strength = 11.7473579[T] (500[MHX_acq_duration = 0.83361792[s]X_domain = 13CX_freq = 125.76529768[MHz]X_offset = 100[ppm]X_points = 32768X_prescans = 4X_resolution = 1.19959034[Hz]X_sweep = 39.3081761[kHz]Irr_domain = 1HIrr_freq = 500.15991521[MHz]Irr_offset = 5.0[ppm]Clipped = FALSEMod_return = 1Scans = 2048Total_scans = 2048
X_90_width = 8.62[us]X_acq_time = 0.83361792[s]X_angle = 30[deg]X_atn = 6.4[dB]X_pulse = 2.87333333[us]Irr_atn_dec = 21.68[dB]Irr_atn_noe = 21.68[dB]Irr_noise = WALTZDecoupling = TRUEInitial_wait = 1[s]Noe = TRUENoe_time = 2[s]Recvr_gain = 60Relaxation_delay = 2[s]Repetition_time = 2.83361792[s]Temp_get = 18.1[dC]
−−−− PROCESSING PARAMETERS −−−−dc_balance : 0 : FALSEsexp : 2.0[Hz] : 0.0[s]fft : 1 : TRUE : TRUEmachinephaseppm
Derived from: 160115_PF_5_13C_CARBON−1.j
44 45
-glucosidase assay of 12p using (p-nitro phenyl -glucopyranoside)
control OD
Compound 12p
336
% of inhibition 0.9 Concentrations (µM)
R1 R2 average C-T IC50 (µg) IC50 (µM)
0.2
0.48 0.45 0.465 43.5 0.4
0.25 0.23 0.24 66 0.82 0.244047619
0.6
0.14 0.13 0.135 76.5 0.8
0.03 0.01 0.02 88
1
0 0.01 0.005 89.5
y = -0.57x + 0.515 R² = 0.9099
-0.1
0
0.1
0.2
0.3
0.4
0.5
0 0.5 1 1.5
46
α-glucosidase 12p-LB plot mM of PNPG (substrate)
[S] 1/s v10 v20 v30 v40 v50
So (mM) 2 4 6
2 0.5 0.465 0.24 0.135 0.08 0.04 Conc: of I (μM) Vo(IU/ml) Vo(IU/ml) Vo(IU/ml) km (-1/km)
4 0.25 0.498 0.32 0.17 0.12 0.07
0.2 0.465 0.498 0.63 0.96 -
1.04167
6 0.166667 0.63 0.41 0.31 0.18 0.09
0.4 0.24 0.32 0.41 2.8 -
0.35714
1/v
1/v10 1/v20 1/v30 1/v40 1/v50
0.6 0.135 0.17 0.31 5.1 -
0.19608
0.5 2.150538 4.166667 7.407407 12.5 25
0.8 0.08 0.12 0.18 7.2 -
0.13889
0.25 2.008032 3.125 5.882353 8.333333 14.28571
1 0.04 0.07 0.09 10.5 -
0.09524
0.166667 1.587302 2.439024 3.225806 5.555556 11.11111
-5
0
5
10
15
20
25
-0.3 -0.2 -0.1 0 0.1 0.2 0.3 0.4 0.5 0.6
1/[
V]
1/[S]
0.2uM
0.4uM
0.6uM
0.8uM
1uM
-1/Km'
(a)
R² = 0.9979
02468
1012
0 0.2 0.4 0.6 0.8 1
km'
[I]
(b)
47
COS cell cytotoxicity screening
expt-1 O.D 1 O.D 2 avg ctrl-avg
/ctrl *100 or % inhibition
control 0.765 0.723 0.744 0 0 0
50 0.735 0.775 0.755 -0.011 -0.01478 -1.47849
25 0.767 0.783 0.775 -0.031 -0.04106 -4.10596
12.5 0.736 0.742 0.739 0.005 0.006452 0.645161
6.25 0.779 0.788 0.7835 -0.0395 -0.05345 -5.34506
3.125 0.743 0.741 0.742 0.002 0.002553 0.255265
1.56 0.861 0.6 0.7305 0.0135 0.018194 1.819407
0.78 0.685 0.752 0.7185 0.0255 0.034908 3.49076
0.39 0.79 0.688 0.739 0.005 0.006959 0.695894
0.195 0.716 0.704 0.71 0.034 0.046008 4.600812
Expt-2 O.D 1 O.D 2 avg
ctrl-avg
/ctrl *100 or % inhibition
control 1.039 1.259 1.149 0 0 0
50 1.07 1.031 1.0505 0.0985 0.085727 8.572672
25 1.197 1.353 1.275 -0.126 -0.10966 -10.9661
12.5 1.211 1.208 1.2095 -0.0605 -0.05265 -5.26545
6.25 1.073 1.126 1.0995 0.0495 0.043081 4.308094
3.125 1.331 1.143 1.237 -0.088 -0.07659 -7.65883
1.56 1.087 1.042 1.0645 0.0845 0.073542 7.354221
0.78 1 1.073 1.0365 0.1125 0.097911 9.791123
0.39 1.074 1.095 1.0845 0.0645 0.056136 5.613577
0.195 1.089 1.076 1.0825 0.0665 0.057876 5.787641
48
To determine the kinetic parameters a linear regression analysis of 12p against -glucosidase was
obtained (Michalis Menten plot)
We have performed a global non-linear regression analysis for different substrate concentrations
in the presence of various concentrations of the potent mixed inhibitor (12p). Afterwards, the data
was linearized and figured the binding constants (Ki) values. The regression plots were as follows
Substituted furopyridinediones as novel inhibitors of -glucosidase
Chandramohan Bathula,1, § Rajanikanth Mamidala,4, § Chiranjeevi Thulluri,3 Rahul Agarwal,2,
Kunal Kumar Jha,1 Parthapratim Munshi,1 Uma Adepally,3 Ashutosh Singh,2 M.
Thirumalachary4, Subhabrata Sen1, *
1 Department of Chemistry, School of Natural Sciences, Shiv Nadar University, Chithera, Dadri, UP
201314, India
2 Department of Life Science, School of Natural Sciences, Shiv Nadar University, Chithera, Dadri, UP
201314, India
3 Institute of Science and Technology, Jawaharlal Nehru Technological University Hyderabad,
Kukatpally, Hyderabad500085, Telengana, India
4 Department of Chemistry, Jawaharlal Nehru Technological University Hyderabad, Kukatpally,
Hyderabad500085, Telengana, India
Figure 1. Ramachandran Plot of modelled -glucosidase protein
For docking studies the modelled structures, 7a, 9a, 12d, 12p, acarbose and template structure
(PDB ID: 3L4T) were aligned and the centroid coordinates of the complex ligand in the crystal
structure (PDB ID: 3L4T) were used as the centre of the docking site. A grid of 60 x 60 x 60 with
0.375 A0 were constructed around the docking area using Autogrid v 4.2 software.
Docking analyses were done using Autodock v 4.2 software, in which top 10 docked conformation
were taken using Genetic Algorithm. Each docking calculation consisted of 25 x 106 energy
evaluations with 250 population size. A mutation rate of 0.02 and a crossover rate of 0.8 were used
to generate new docking trials.
Docking interactions of 7a, 9a and 12d
Table 1
Compound Name Residues involved in H-
bond
Residues involved in other
non-bonded interactions
Compound 7a ARG 193, ARG 194, HIS 586 TRP 288, ASP 316, ILE 353,
TRP 393, TRP 428, ASP 430,
MET 431, PHE 437, ARG
512, TRP 525, ASP 528
Compound 9a ARG 193, ASP 528 ASP 194, TRP 393, ALA 467
Compound 12d SER 435, ARG 512 ARG 193, ASP 194, LEU
195, ALA 196, PHE 437,
ALA 467, ASP 528, LEU 562
Note: Highlighted residues are important for alpha-glucosidase activity.
Alpha glucosidase assay screening using (p-Nitro phenyl alpha D- glucopyranoside)
control OD
Compound 12a 281.31 % of inhibition 0.58
Concentrations (µg) R1 R2 average C-T Ic 50 (µg) Ic 50 (µM)
10 0.871 0.798 0.8345
30 1.656 1.864 1.76
50 1.505 2.313 1.909
70 2.914 3.322 3.118
100 3.691 3.613 3.652
Compound 12c 321.25
Concentrations (µg) R1 R2
2 0.24 0.21 0.225 35.5
4 0.132 0.132 0.132 44.8 7.427835 23.12
6 0.096 0.075 0.0855 49.45
8 0.068 0.069 0.0685 51.15
10 0.061 0.065 0.063 51.7
Compound 12b 311.33
Concentrations (µg) R1 R2
2 1.454 1.406 1.43
4 2.8 2.454 2.627
6 2.715 2.596 2.6555
8 2.588 2.465 2.5265
10 2.666 2.735 2.7005
Compound 12e 332.15
Concentrations (µg) R1 R2
2 0.171 0.115 0.143 43.7
4 0.097 0.147 0.122 45.8 13.68889 41.21297272
6 0.122 0.104 0.113 46.7
8 0.137 0.084 0.1105 46.95
10 0.079 0.128 0.1035 47.65
NOT ACTIVE
NOT ACTIVE
y = -0.0194x + 0.2311 R² = 0.8284
0
0.05
0.1
0.15
0.2
0.25
0 2 4 6 8 10
y = -0.0045x + 0.1456 R² = 0.8791
0
0.02
0.04
0.06
0.08
0.1
0.12
0.14
0.16
0 2 4 6 8 10
Compound 12g 303
Concentrations (µg) R1 R2
2 0.573 1.405
4 0.895 0.965 0.93
6 1.375 1.546 1.4605
8 2.435 2.46 2.4475
10 2.64 2.628 2.634
Compound 12i 297
Concentrations (µg) R1 R2
2 0.681 0.784 0.7325
4 1.797 1.784 1.7905
6 2.876 2.804 2.84
8 2.874 2.932 2.903
10 2.88 2.96 2.92
Compound 12j 313
Concentrations (µg) R1 R2
2 0.43 0.449 0.4395
4 0.906 0.873 0.8895
6 1.753 2.337 2.045
8 2.311 2.27 2.2905
10 2.4 2.36 2.38
Compound 12s 366
Concentrations (µg) R1 R2
2 0.37 0.35 0.36 22.0
4 0.36 0.33 0.345 23.5 11.2 30.6010929
6 0.29 0.24 0.265 31.5
8 0.21 0.22 0.215 36.5
10 0.2 0.2 0.2 38.0
NOT ACTIVE
NOT ACTIVE
NOT ACTIVE
y = -0.0225x + 0.412 R² = 0.9449
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0 2 4 6 8 10
Compound 12r 336
Concentrations (µg) R1 R2
2 0.396 0.339 0.3675
4 0.798 0.504 0.651
6 1.248 1.256 1.252
8 1.872 1.913 1.8925
10 2.102 2.225 2.1635
Compound 12u 380
Concentrations (µg) R1 R2
2 0.589 0.558 0.5735
4 1.214 0.998 1.106
6 2.141 2.233 2.187
8 2.729 2.876 2.8025
10 3.066 3.005 3.0355
Compound 12v 343
Concentrations (µg) R1 R2
2 0.672 0.744 0.708
4 1.282 1.707 1.4945
6 2.071 2.713 2.392
8 2.921 2.888 2.9045
10 3.1 2.886 2.993
Compound 12k 253
Concentrations (µg) R1 R2
2 0.44 0.4 0.42 16.0
4 0.36 0.38 0.37 21.0 13.50719 53.38811954
6 0.28 0.32 0.3 28.0
8 0.25 0.26 0.255 32.5
10 0.2 0.2 0.2 38.0
NOT ACTIVE
NOT ACTIVE
NOT ACTIVE
y = -0.0278x + 0.4755 R² = 0.9962
0
0.1
0.2
0.3
0.4
0.5
0 2 4 6 8 10
Compound 12d 285.25
Concentrations (µg) R1 R2 C-T Ic 50 (µg) Ic 50 (µM)
2 0.18 0.2 0.19 50
4 0.12 0.132 0.126 62 0.575916 2.02
6 0.073 0.075 0.074 75
8 0.04 0.06 0.05 88
10 0.014 0.02 0.017 96
Compound 12f 332.15
Concentrations (µg) R1 R2
2 0.24 0.21 0.225 39
4 0.155 0.168 0.1615 50.1 3.5311 10.63
6 0.096 0.075 0.0855 69
8 0.068 0.069 0.0685 88
10 0.061 0.065 0.063 96
Compound 12h 289.23
Concentrations (µg) R1 R2
2 0.24 0.21 0.225 50.5
4 0.132 0.132 0.132 59.8 0.314433 1.09
6 0.096 0.075 0.0855 64.45
8 0.068 0.069 0.0685 66.15
10 0.061 0.065 0.063 66.7
y = -0.0211x + 0.218 R² = 0.9641
0
0.05
0.1
0.15
0.2
0 5 10 15
y = -0.0209x + 0.2458 R² = 0.8766
0
0.05
0.1
0.15
0.2
0.25
0 5 10 15
y = -0.0194x + 0.2311 R² = 0.8284
0
0.05
0.1
0.15
0.2
0.25
0 5 10 15
Compound 12l 298.25
Concentrations (µg) R1 R2
2 0.28 0.27 0.275 47.5
4 0.17 0.19 0.18 57 1.057432 3.55
6 0.096 0.075 0.0855 66.45
8 0.068 0.069 0.0685 68.15
10 0.04 0.03 0.035 71.5
Compound 12m 292.29
Concentrations (µg) R1 R2
2 0.24 0.21 0.225 50
4 0.132 0.132 0.132 62 0.628272 2.15
6 0.096 0.075 0.0855 75
8 0.068 0.069 0.0685 88
10 0.061 0.065 0.063 96
Compound 12n 243.22
Concentrations (µg) R1 R2
2 0.11 0.14 0.125 45.5
4 0.08 0.06 0.07 51 0.227642 0.94
6 0.03 0.05 0.04 54
8 0.01 0.02 0.015 56.5
10 0.01 0.01 0.01 57
y = -0.0296x + 0.3063 R² = 0.9134
0
0.05
0.1
0.15
0.2
0.25
0.3
0 5 10 15
y = -0.0194x + 0.2311 R² = 0.8284
0
0.05
0.1
0.15
0.2
0.25
0 5 10 15
y = -0.0143x + 0.1375 R² = 0.9096
-0.02
0
0.02
0.04
0.06
0.08
0.1
0.12
0.14
0 5 10 15
Compound 12o 259.28
Concentrations (µg) R1 R2
2 0.24 0.21 0.225 39
4 0.155 0.168 0.1615 50.1 2.669856 10.30
6 0.096 0.075 0.0855 69
8 0.068 0.069 0.0685 88
10 0.061 0.065 0.063 96
Compound 12q 335.38
Concentrations (µg) R1 R2
2 0.13 0.15 0.14 44
4 0.07 0.08 0.075 50.5 2.8125 8.39
6 0.04 0.05 0.045 53.5
8 0.01 0.02 0.015 56.5
10 0.01 0.01 0.01 57
Compound 12t 520.18
Concentrations (µg) R1 R2
2 0.32 0.35 0.335 24.5
4 0.28 0.25 0.265 31.5 45.05814 86.62
6 0.2 0.19 0.195 38.5
8 0.12 0.14 0.13 45 86
10 0.08 0.11 0.095 48.5
y = -0.0273x + 0.2715 R² = 0.9492
0
0.05
0.1
0.15
0.2
0.25
0 2 4 6 8 10
y = -0.016x + 0.153 R² = 0.9038
-0.02
0
0.02
0.04
0.06
0.08
0.1
0.12
0.14
0.16
0 5 10 15
y = -0.0308x + 0.3885 R² = 0.987
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0 5 10 15
control OD
Compound 2p 336 % of inhibition 0.9
Concentrations (µM) R1 R2 average C-T Ic 50 (µg) Ic 50 (µM)
0.2 0.48 0.45 0.465 43.5
0.4 0.25 0.23 0.24 66 0.82 0.244047619
0.6 0.14 0.13 0.135 76.5
0.8 0.03 0.01 0.02 88
1 0 0.01 0.005 89.5
y = -0.57x + 0.515 R² = 0.9099
-0.1
0
0.1
0.2
0.3
0.4
0.5
0 0.5 1 1.5