ars.els-cdn.com€¦ · Web viewSupplementary Table 1. Supplier information, Glide XP GScores,...
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Supplementary Materials: Supplementary Table 1. Supplier information, Glide XP GScores, and ranking for each of the assessed compounds. Supplementary Table 2. Suppression of constitutive activity of RORγ by virtually screened compounds and reference ligands. Supplementary Figure S1. 2D structures of the first round hit compounds from virtual screening. Supplementary Figure S2: Single point activity result for the screened compounds at 50 μM using the AlphaScreen assay. Supplementary Figure S3: Predicted binding modes of additional ligands with the LBD. Supplementary Figure S4: Dose-response curves for 1
Transcript of ars.els-cdn.com€¦ · Web viewSupplementary Table 1. Supplier information, Glide XP GScores,...
Supplementary Materials:
Supplementary Table 1. Supplier information, Glide XP GScores, and ranking for
each of the assessed compounds.
Supplementary Table 2. Suppression of constitutive activity of RORγ by virtually
screened compounds and reference ligands.
Supplementary Figure S1. 2D structures of the first round hit compounds from
virtual screening.
Supplementary Figure S2: Single point activity result for the screened compounds at
50 μM using the AlphaScreen assay.
Supplementary Figure S3: Predicted binding modes of additional ligands with the
LBD.
Supplementary Figure S4: Dose-response curves for suppression of constitutive
activity of RORγ by T1317, 7j, 8c, 8k, and 8p in the AlphaScreen and reporter gene
assays.
Supplementary Figure S5: 1H-NMR and 13C-NMR spectra of compounds 6f, 7a, 7k,
8a, 8c, 8i, 8k, and 8s.
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Supplementary Table 1. Supplier information, Glide XP GScores, and ranking for each of the tested compounds.
Compound SPECS ID XP-GScore Ranking MW AlogPs1 AR-318/42871695 -9.09 23 364.80 2.94s2 AK-968/15254261 -9.06 24 457.93 3.96s3 AG-205/11218162 -10.54 4 559.41 5.35s4 AG-690/15438001 -9.29 15 460.50 3.37s5 AN-919/40868728 -9.26 18 565.66 4.15s6 AG-690/40751044 -10.67 2 539.57 5.45s7 AK-968/41170314 -9.96 8 531.51 5.10s8 AH-487/42145080 -9.18 20 492.57 3.36s9 AN-989/40872722 -10.61 3 540.49 2.34s10 AK-968/40642622 -9.61 10 380.82 3.82s11 AK-968/40709644 -9.37 13 406.84 2.90s12 AN-329/10088022 -9.12 22 480.55 6.95s13 AO-476/43362644 -9.15 21 463.96 3.00s14 AN-919/41152740 -9.51 12 582.65 3.25s15 AN-465/41377374 -9.27 17 378.79 3.43s16 AN-919/41439252 -9.35 14 472.62 4.06s17 AF-399/41692703 -9.80 9 590.67 2.70s18 AG-650/41069241 -10.35 6 456.40 0.53s19 AO-022/43453007 -9.29 16 450.89 3.37s20 AO-022/43454460 -10.43 5 476.50 3.65s21 AN-465/42784379 -10.73 1 430.95 4.18s22 AN-465/43369996 -10.08 7 390.88 3.51s23 AP-970/42224548 -9.58 11 418.31 5.07s24 AQ-390/43363961 -9.24 19 431.49 1.54
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Supplementary Table 2. Suppression of constitutive activity of RORγ by virtually screened compounds and reference ligands.
Cmpd
Alphascreen
Luciferase
RORγ-LBD Gal4-RORγ-LBD full-length-RORγ
IC50 (μM)IC50
(μM)Inhibitory rate at
10μM(%)IC50
(μM)Inhibitory rate at
10μM(%)T1317 2.50 ± 0.87 0.54 82 3.50 76
UA 9.41 ± 1.07 0.13 76 1.06 63SR221
13.54 ± 0.01 0.43 79 0.42 44
s1 — — <10 — <10s2 56.64±7.74 <30 <10s3 — — <10 — <10s4 20.27±6.26 11.84 79 7.56 77s5 11.14±0.24 — <40 — <20s6 11.64±1.19 — <20 — <10s7 — — <10 — <10s8 15.17±0.77 — <20 — <10s9 2.44±0.56 — <30 — <20
s10168.27±8.1
1— <10 — <10
s11 81.69±8.31 — <20 — <10s12 37.54±1.41 — <10 — <10s13 — — <30 — <30s14 46.87±9.37 — <30 — <10s15 — — <20 — <10s16 18.27±0.34 — <10 — <10
s17142.21±8.4
8— <10 — <10
s18 — — <10 — <10s19 9.05±2.00 — <10 — <10s20 — — <10 — <10s21 — — <10 — <10s22 — — <10 — <10s23 — — <10 — <10s24 — — <50 5.15 42
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Supplementary Figure S1. 2D structures of the first round hit compounds from virtual screening.
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Supplementary Figure S2: Single point activity result for the screened compounds at 50 μM using the AlphaScreen assay.
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Supplementary Figure S3: Predicted binding modes of additional ligands with the RORγ LBD. (A) RORγ-6a, (B) RORγ-6d, (C) RORγ-7a, (D) RORγ-7k, (E) RORγ-7m, (F) RORγ-8g.
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Supplementary Figure S4: Dose-response curves for suppression of constitutive activity of RORγ by T1317, 7j, 8c, 8k, and 8p in the AlphaScreen and reporter gene assays. Demonstrated curves are from one of the three experiments. For the Alphascreen assay, the IC50 values were 2.01, 3.59, 73.25, 5.88 μM; for Gal4-RORγLBD, the IC50 values were 0.54, 0.14, 0.04, 0.05, 0.09 μM; and for full length RORγ, the IC50 values were 3.5, 0.15, 0.05, 0.02, and 0.08 μM respectively.
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Supplementary Figure S5: 1H-NMR and 13C-NMR spectra of compounds 6f, 7a, 7k, 8a, 8c, 8i, 8k, and 8s.
1-ethyl-N-(isoquinolin-7´-yl)-2-oxo-1,2-dihydrobenzo[cd]indole-6-sulfonamide(6f).
200 180 160 140 120 100 80 60 40 20 0 ppm
14.47
35.50
104.85
114.67
120.78
124.80
124.85
125.76
126.17
126.98
128.22
128.90
129.39
129.91
131.69
132.64
134.77
137.32
142.79
144.42
152.46
167.57
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1-ethyl-N-(4´-morpholinophenyl)-2-oxo-1,2-dihydrobenzo[cd]indole-6-sulfonamide
(7a).
200 180 160 140 120 100 80 60 40 20 0 ppm
14.5
4
35.5
1
49.3
2
66.8
8
104.
90
116.
39123.
45125.
06125.
65126.
02126.
88129.
20129.
54130.
31131.
34133.
95143.
91149.
16
167.
63
9
(E)-1-ethyl-2-oxo-N-(4´-(phenyldiazenyl)phenyl)-1,2-dihydrobenzo[cd]indole-6-
sulfonamide(7k).
200 180 160 140 120 100 80 60 40 20 0 ppm
14.01
14.59
35.28
104.41
119.86
122.78
124.32
128.57
131.25
131.51
134.04
141.25
144.24
148.68
152.77
167.30
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Tert-butyl(3´-(1-ethyl-2-oxo-1,2-dihydrobenzo[cd]indole-6-
sulfonamido)phenyl)carbamate(8a).
11
200 180 160 140 120 100 80 60 40 20 0 ppm
14.5
4
28.9
8
35.5
2
79.9
8
104.
8511
0.00
113.
7711
4.38
124.
9612
5.76
126.
0712
6.96
128.
8313
0.03
130.
2013
1.51
134.
4813
8.83
141.
1214
4.16
153.
44
167.
66
N-(4´-acetylphenyl)-1-ethyl-2-oxo-1,2-dihydrobenzo[cd]indole-6-sulfonamide(8c).
12
200 180 160 140 120 100 80 60 40 20 0 ppm
13.9
9
26.9
5
35.2
6
104.
39
119.
2812
4.37
124.
5512
4.76
125.
6312
6.73
128.
6812
9.73
130.
0713
1.15
133.
8113
8.44
138.
7014
4.12
167.
30
197.
57
N-(2´-(tert-butyl)phenyl)-1-ethyl-2-oxo-1,2-dihydrobenzo[cd]indole-6-sulfonamide
(8i).
13
200 180 160 140 120 100 80 60 40 20 0 ppm
14.0
5
32.2
835
.31
35.9
0
104.
5912
4.96
125.
5312
5.71
126.
6512
6.72
127.
9212
8.31
129.
9213
0.72
130.
7913
1.90
132.
9413
4.99
143.
6214
8.58
167.
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N-(3´,4´dimethoxyphenyl)-1-ethyl-2-oxo-1,2-dihydrobenzo[cd]indole-6-sulfonamide
14
(8k).
200 180 160 140 120 100 80 60 40 20 0 ppm
14.0
8
35.2
9
56.2
956.6
1
104.
49
113.
56114.
47125.
00125.
58126.
70129.
12130.
08130.
98131.
20133.
68143.
83147.
09149.
74
167.
39
N-(2´,5´-dichlorobenzyl)-1-ethyl-2-oxo-1,2-dihydrobenzo[cd]indole-6-sulfonamide
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(8s).
200 180 160 140 120 100 80 60 40 20 0 ppm
14.1
3
35.2
5
42.2
1
104.
32
124.
9712
5.40
126.
5312
8.83
130.
5013
0.58
130.
7313
2.42
133.
0913
5.99
143.
54
167.
48
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