Supplementary Materials for
Identification of a small-molecule ligand that activates the neuropeptide
receptor GPR171 and increases food intake
Jonathan H. Wardman, Ivone Gomes, Erin N. Bobeck, Jennifer A. Stockert,
Abhijeet Kapoor, Paola Bisignano, Achla Gupta, Mihaly Mezei, Sanjai Kumar,
Marta Filizola, Lakshmi A. Devi*
*Corresponding author. Email: [email protected]
Published 31 May 2016, Sci. Signal. 9, ra55 (2016)
DOI: 10.1126/scisignal.aac8035
The PDF file includes:
Fig. S1. Binding of b-LEN–derived peptides and ligands of P2Y12 or P2Y14 to
GPR171.
Fig. S2. Sequence alignment between mouse GPR171 and P2Y12.
Fig. S3. Homology model of GPR171 superimposed with the P2Y12 crystal structure.
Fig. S4. Surface receptor abundance of mutant GPR171 constructs expressed in CHO
cells.
Fig. S5. Selectivity profile MS0015203.
Fig. S6. Effect of acute administration of MS0015203 on food intake in mice.
Fig. S7. Effect of GPR171 knockdown on receptor abundance in hypothalamus.
Table S1. Structures of compounds representative of each cluster.
Table S2. Displacement of radiolabeled b-LEN by a compound with chemical
similarity to MS0015203.
www.sciencesignaling.org/cgi/content/full/9/430/ra55/DC1
fig. S1. Binding of b-LEN–derived peptides and ligands of P2 Y12 or P2Y14 to GPR171. (A) Displacement of [125I]Tyr-b-LEN binding (3 nM) with b-LEN (LENPSPQAPARRLLPP), NPSPQAPARRLLPP, SPQAPARRLLPP, QAPARRLLPP, PARRLLPP, RRLLPP, RLLPP, LLPP, LPP (0-10 µM) using rat hypothalamic membranes. Binding in the absence of cold ligands is taken as 100%. (B) Displacement of [125I]Tyr-b-LEN binding (3 nM) in rat hypothalamic membranes by b-LEN (10 µM), P2Y ligands (10 µM) and dicarboxylic acid receptor ligands (10 µM). Data in A and B represent means ± SE (n=3-6).
fig. S2. Sequence alignment between mouse GPR171 and P2Y 12. Identical residues between GPR171 and P2Y12 that are located in the crystallographic binding pocket of the P2Y12 receptor are highlighted red. Annotations above conserved residues indicate Ballesteros-Weinstein generic numbering scheme for class A GPCRs, EL2 represents extracellular loop 2. Black stars indicate the most conserved residues in each transmembrane (TM) helix across the entire class A GPCR family. IL#, intracellular loop number; EL#, extracellular loop number.
fig. S3. Homology model of GPR171 superimposed with the P2Y 12 crystal structure. The homology model of GPR171 (gray) is constructed from PDB ID: 4PXZ of P2Y12 (cyan). (A) P2Y12 residue side chains within 3.5 Å of the crystal ligand (2MeSADP, shown as spheres on the left or sticks in the inset), are shown as lines. Residues are labeled using one-letter amino acid code and their sequence number, with the Ballesteros-Weinstein’s generic number reported as superscript. Residue side chains that are different in GPR171 are also shown as sticks, and labeled in red. (B) Predicted binding mode of the agonist MS0015203 (yellow) superimposed to the crystal ligand 2MeSADP (orange).
fig. S4. Surface receptor abundance of mutant GPR171 constru cts expressed in CHO cells. The amount of receptor at the cell surface receptor for DYKDDDDK-tagged mouse GPR171 (mGPR171) or the indicated mutants was determined by ELISA using antibodies that recognize the DYKDDDDK tag. Surface receptor abundance data are represented as the overall O.D. 490 nm values observed after subtraction of nonspecific binding and are the mean ± SEM of 2 independent experiments in triplicate.
fig. S5. Selectivity profile MS0015203. (A) The selectivity profile of MS15203 (10 µM) was carried out by Eurofins Cerep Panlabs using LeadProfileScreen 2 that included 80 transmembrane and soluble receptors, as well as ion channels and monoamine transporters. (B) Left: Surface abundance of mouse Gal1R (mGal1R) and mouse P2Y12 in CHO cells expressing DYKDDDDK-tagged versions of the receptor. Cell surface receptor abundance was determined by ELISA with antibodies to the DYKDDDDK tag. Surface receptor abundance data are represented as the overall O.D. 490 nm values observed after subtraction of nonspecific binding. Middle: The effect of M617 (Gal1R agonist) and MS0015203 (0-10 µM) and of M617 (100 nM) together with MS0015203 (0-10 µM) on cAMP concentrations in CHO cells expressing mGal1R. Right: The effect of 2MeSADP (P2Y12 agonist) and MS0015203 (0-100 µM) and of 2MeSADP (100 nM) together with MS0015203 (0-100 µM) on cAMP concentrations in CHO cells expressing mP2Y12. Data in B represent the mean ± SEM of 2 independent experiments in triplicate.
fig. S6. Effect of acute administration of MS0015203 on food intake in mice. (A) Food intake on an hour-by-hour basis in fasted mice administered with MS0015203 (3 mg/kg; i.p., n=6) or vehicle (6% DMSO; n=5). **p< 0.01 vehicle vs. MS0015203 (Two-Way ANOVA with Bonferroni’s post-hoc comparisons; Interaction: F(3,27)=0.4844, p=0.6959; Time: F(3,27)=19.66, p<0.0001; Drug: F(1,9)=6.639, p=0.0299; Subjects (matching): F(9,27)=1.155, p<0.3614).
fig. S7. Effect of GPR171 knockdown on receptor ab undance in hypothalamus. (A) Western blot for GPR171 in lysates (~15 µg protein) from individual dorsal hypothalami of mice administered control virus (CV; n=4) or GPR171-knockdown virus (KDV; n=2 for 2 weeks after viral administration; n=2 for 3 weeks after viral administration) into PVN using anti-rabbit GPR171 antibody (green; 1:1000) and anti-mouse tubulin antibodies (red; 1:50,000). *p<0.05; One-Way ANOVA. (B) Western blot for GPR171 in lysates (~15 µg protein) from individual ventral hypothalami of mice administered control virus (CV; n=4) or GPR171-knockdown virus (KDV; n=2 for 2 weeks after viral administration; n=2 for 3 weeks after viral administration).
Table S1. Structures of compounds representative of each cluster. 2D structures of 502
representative compounds resulting from clustering of top-scoring 5842 compounds identified by
virtual screening for binding to GPR171. The name of the compound representative of the group, its
rank in binding based on energy score, and the energy score are included.
Cluster
id.
Structure
Name
Cluster
id.
Structure
Name
1
MS0014904
Rank:16
Score:
-38.26
2
MS0014903
Rank:793
Score:-25.48
3
MS0013451
Rank:51
Score:
-34.71
4
MS0019262
Rank:3096
Score:-19.12
5
MS0014370
Rank:546
Score:
-26.83
6
MS0019619
Rank:150
Score:-31
7
MS0015577
Rank:5804
Score:
-15.05
8
MS0015203
Rank:43
Score:-35.25
9
MS0014024
Rank:1107
Score:
-24.13
10
MS0015515
Rank:324
Score:-28.53
11
MS0012626
Rank:151
Score:
-30.98
12
MS0018906
Rank:497
Score:-27.17
13
MS0015921
Rank:1096
Score:
-24.16
14
MS0022421
Rank:54
Score:-34.41
15
MS0021249
Rank:340
Score:
-28.39
16
MS0013658
Rank:100
Score:-32.27
17
MS0012671
Rank:1055
Score:
-24.34
18
MS0012772
Rank:1373
Score:-23.18
19
MS0021865
Rank:46
Score:
-34.93
20
MS0022303
Rank:4598
Score:-16.79
21
MS0018844
Rank:41
Score:
-35.49
22
MS0020105
Rank:1302
Score:-23.42
23
MS0021152
Rank:3626
Score:
-18.22
24
MS0015240
Rank:4211
Score:-17.34
25
MS0014045
Rank:246
Score:-29.4 26
MS0012092
Rank:1908
Score:-21.66
27
MS0016191
Rank:341
Score:
-28.36
28
MS0022423
Rank:1444
Score:-22.94
29
MS0020188
Rank:3375
Score:
-18.64
30
MS0015547
Rank:1736
Score:-22.09
31
MS0015008
Rank:642
Score:
-26.25
32
MS0015460
Rank:1248
Score:-23.61
33
MS0017272
Rank:3261
Score:
-18.82
34
MS0012457
Rank:104
Score:-32.2
35
MS0019390
Rank:604
Score:
-26.46
36
MS0015601
Rank:199
Score:-30.18
37
MS0018894
Rank:3735
Score:
-18.05
38
MS0022084
Rank:2749
Score:-19.72
39
MS0015541
Rank:5793
Score:
-15.07
40
MS0016247
Rank:5
Score:-41.86
41
MS0016513
Rank:1255
Score:
-23.58
42
MS0012840
Rank:243
Score:-29.45
43
MS0013767
Rank:62
Score:
-33.83
44
MS0016790
Rank:570
Score:-26.69
45
MS0012201
Rank:289
Score:
-28.92
46
MS0016616
Rank:934
Score:-24.86
47
MS0022681
Rank:306
Score:
-28.72
48
MS0021116
Rank:388
Score:-27.83
49
MS0020482
Rank:297
Score:
-28.85
50
MS0022643
Rank:1693
Score:-22.18
51
MS0015372
Rank:1729
Score:-22.1 52
MS0015917
Rank:34
Score:-36.08
53
MS0020548
Rank:2254
Score:
-20.78
54
MS0019723
Rank:351
Score:-28.24
55
MS0014639
Rank:709
Score:
-25.87
56
MS0015081
Rank:8
Score:-40.56
57
MS0021830
Rank:203
Score:
-30.11
58
MS0011965
Rank:89
Score:-32.5
59
MS0020337
Rank:144
Score:
-31.2
60
MS0022537
Rank:643
Score:-26.25
61
MS0014095
Rank:311
Score:
-28.67
62
MS0012755
Rank:3590
Score:-18.29
63
MS0021012
Rank:64
Score:
-33.79
64
MS0012187
Rank:1120
Score:-24.11
65
MS0015197
Rank:2447
Score:
-20.35
66
MS0021081
Rank:321
Score:-28.59
67
MS0012011
Rank:3008
Score:
-19.29
68
MS0017530
Rank:18
Score:-38.05
69
MS0018456
Rank:229
Score:
-29.62
70
MS0015400
Rank:164
Score:-30.76
71
MS0020163
Rank:146
Score:
-31.14
72
MS0021853
Rank:159
Score:-30.9
73
MS0014947
Rank:31
Score:
-36.61
74
MS0014822
Rank:107
Score:-32.18
75
MS0012585
Rank:5837
Score:
-15.01
76
MS0011872
Rank:679
Score:-26.06
77
MS0014428
Rank:1304
Score:-23.4 78
MS0014274
Rank:927
Score:-24.88
79
MS0014609
Rank:1698
Score:
-22.18
80
MS0012536
Rank:5529
Score:-15.42
81
MS0016494
Rank:2221
Score:
-20.84
82
MS0021119
Rank:1632
Score:-22.38
83
MS0020190
Rank:525
Score:
-26.99
84
MS0014707
Rank:857
Score:-25.18
85
MS0021049
Rank:5589
Score:
-15.34
86
MS0019339
Rank:2572
Score:-20.04
87
MS0021059
Rank:496
Score:
-27.17
88
MS0022252
Rank:1014
Score:-24.47
89
MS0013522
Rank:1884
Score:
-21.71
90
MS0018766
Rank:20
Score:-37.91
91
MS0014591
Rank:71
Score:
-33.44
92
MS0013493-
imp1
Rank:1181
Score:-23.88
93
MS0021951
Rank:1317
Score:
-23.36
94
MS0015882
Rank:526
Score:-26.98
95
MS0019128
Rank:180
Score:
-30.59
96
MS0014722
Rank:909
Score:-24.96
97
MS0021713
Rank:3539
Score:
-18.38
98
MS0020571
Rank:2222
Score:-20.84
99
MS0017157
Rank:666
Score:
-26.11
100
MS0020954
Rank:4084
Score:-17.53
101
MS0021010
Rank:2441
Score:
-20.37
102
MS0019123
Rank:49
Score:-34.8
103
MS0018296
Rank:1114
Score:
-24.11
104
MS0018654
Rank:128
Score:-31.67
105
MS0123522
Rank:3902
Score:
-17.82
106
MS0021912
Rank:309
Score:-28.68
107
MS0017289
Rank:1588
Score:
-22.53
108
MS0011629
Rank:4596
Score:-16.79
109
MS0123924
Rank:4853
Score:
-16.43
110
MS0013115
Rank:972
Score:-24.65
111
MS0022208
Rank:19
Score:
-37.97
112
MS0021607
Rank:3912
Score:-17.8
113
MS0021391
Rank:214
Score:
-29.96
114
MS0019137
Rank:13
Score:-38.49
115
MS0020515
Rank:3348
Score:
-18.68
116
MS0021505
Rank:50
Score:-34.75
117
MS0020534
Rank:118
Score:
-31.84
118
MS0020533
Rank:133
Score:-31.58
119
MS0015484
Rank:264
Score:
-29.19
120
MS0019925
Rank:622
Score:-26.39
121
MS0016654
Rank:2091
Score:
-21.15
122
MS0019411
Rank:78
Score:-32.86
123
MS0015641
Rank:1132
Score:
-24.08
124
MS0019868
Rank:3326
Score:-18.71
125
MS0018813
Rank:1056
Score:
-24.34
126
MS0019449
Rank:467
Score:-27.36
127
MS0020578
Rank:1377
Score:
-23.15
128
MS0013120
Rank:2662
Score:-19.88
129
MS0014696
Rank:4164
Score:
-17.42
130
MS0018017
Rank:1479
Score:-22.83
131
MS0019906
Rank:2962
Score:
-19.36
132
MS0021451
Rank:1018
Score:-24.46
133
MS0021722
Rank:2097
Score:
-21.14
134
MS0012783
Rank:3012
Score:-19.27
135
MS0021382
Rank:255
Score:-29.3 136
MS0021943
Rank:1470
Score:-22.86
137
MS0011604
Rank:4984
Score:
-16.22
138
MS0015067
Rank:5101
Score:-16.04
139
MS0022054
Rank:123
Score:
-31.78
140
MS0016679
Rank:748
Score:-25.65
141
MS0013329
Rank:795
Score:
-25.47
142
MS0012503
Rank:1971
Score:-21.52
143
MS0020267
Rank:3285
Score:
-18.79
144
MS0012244
Rank:4740
Score:-16.6
145
MS0018843
Rank:809
Score:
-25.39
146
MS0018450
Rank:147
Score:-31.14
147
MS0020288
Rank:1439
Score:
-22.98
148
MS0016078
Rank:981
Score:-24.6
149
MS0019591
Rank:283
Score:-
28.98
150
MS0016252
Rank:2123
Score:-21.06
151
MS0021991
Rank:1265
Score:
-23.55
152
MS0012240
Rank:2655
Score:-19.89
153
MS0018663
Rank:101
Score:
-32.26
154
MS0013620
Rank:982
Score:-24.6
155
MS0019545
Rank:522
Score:
-26.99
156
MS0019836
Rank:3381
Score:-18.62
157
MS0013741
Rank:1937
Score:
-21.59
158
MS0015954
Rank:1165
Score:-23.92
159
MS0013706
Rank:6
Score:
-40.94
160
MS0019653
Rank:664
Score:-26.12
161
MS0017260
Rank:9
Score:
-39.54
162
MS0017782
Rank:1057
Score:-24.34
163
MS0012794
Rank:30
Score:
-36.65
164
MS0017341
Rank:335
Score:-28.42
165
MS0015556
Rank:218
Score:
-29.86
166
MS0019849
Rank:1718
Score:-22.13
167
MS0017540
Rank:3178
Score:
-18.97
168
MS0015193
Rank:7
Score:-40.81
169
MS0012854
Rank:63
Score:
-33.83
170
MS0018346
Rank:112
Score:-32.03
171
MS0014869
Rank:14
Score:
-38.32
172
MS0020999
Rank:3749
Score:-18.03
173
MS0019149
Rank:69
Score:
-33.66
174
MS0019186
Rank:1053
Score:-24.35
175
MS0015102
Rank:4634
Score:
-16.75
176
MS0018820
Rank:40
Score:-35.53
177
MS0011733
Rank:2457
Score:
-20.32
178
MS0013809
Rank:869
Score:-25.13
179
MS0017708
Rank:2933
Score:-19.4 180
MS0016527
Rank:91
Score:-32.48
181
MS0015425
Rank:4414
Score:
-17.04
182
MS0017600
Rank:847
Score:-25.23
183
MS0019336
Rank:97
Score:
-32.35
184
MS0012491
Rank:1804
Score:-21.91
185
MS0020953
Rank:1229
Score:
-23.66
186
MS0015315
Rank:698
Score:-25.95
187
MS0022206
Rank:1015
Score:
-24.47
188
MS0022178
Rank:1524
Score:-22.71
189
MS0019378
Rank:271
Score:
-29.11
190
MS0020253
Rank:5480
Score:-15.48
191
MS0019721
Rank:823
Score:
-25.32
192
MS0014073
Rank:27
Score:-37.18
193
MS0014444
Rank:2113
Score:
-21.09
194
MS0016755
Rank:1323
Score:-23.34
195
MS0019435
Rank:261
Score:
-29.23
196
MS0017710
Rank:1336
Score:-23.31
197
MS0014824
Rank:891
Score:
-25.03
198
MS0012358
Rank:840
Score:-25.25
199
MS0019207
Rank:4804
Score:-16.5 200
MS0017384
Rank:2005
Score:-21.41
201
MS0013047
Rank:44
Score:
-35.21
202
MS0015013
Rank:2852
Score:-19.52
203
MS0019954
Rank:1311
Score:
-23.38
204
MS0022076
Rank:1093
Score:-24.16
205
MS0020392
Rank:59
Score:
-33.87
206
MS0021670
Rank:61
Score:-33.83
207
MS0012724
Rank:1088
Score:
-24.18
208
MS0016129
Rank:663
Score:-26.13
209
MS0020988
Rank:1184
Score:
-23.87
210
MS0020075
Rank:1973
Score:-21.49
211
MS0019582
Rank:252
Score:
-29.32
212
MS0014445
Rank:3183
Score:-18.97
213
MS0015249
Rank:162
Score:
-30.84
214
MS0011890
Rank:600
Score:-26.51
215
MS0021883
Rank:294
Score:
-28.89
216
MS0012411
Rank:3927
Score:-17.78
217
MS0020543
Rank:2783
Score:
-19.64
218
MS0124232
Rank:2028
Score:-21.35
219
MS0015398
Rank:245
Score:
-29.41
220
MS0021996
Rank:17
Score:-38.13
221
MS0019119
Rank:810
Score:
-25.39
222
MS0015776
Rank:2034
Score:-21.34
223
MS0015775
Rank:4949
Score:
-16.26
224
MS0015531
Rank:5598
Score:-15.33
225
MS0014910
Rank:4114
Score:
-17.49
226
MS0011500
Rank:1585
Score:-22.53
227
MS0020436
Rank:2342
Score:
-20.59
228
MS0022664
Rank:4941
Score:-16.28
229
MS0013117
Rank:72
Score:
-33.31
230
MS0022728
Rank:422
Score:-27.63
231
MS0019333
Rank:3519
Score:-18.4 232
MS0022434
Rank:92
Score:-32.48
233
MS0016933
Rank:441
Score:
-27.46
234
MS0013049
Rank:201
Score:-30.17
235
MS0015480
Rank:1001
Score:
-24.53
236
MS0022083
Rank:1459
Score:-22.89
237
MS0016902
Rank:1138
Score:
-24.03
238
MS0019522
Rank:168
Score:-30.72
239
MS0014068
Rank:2229
Score:
-20.83
240
MS0011997
Rank:5062
Score:-16.1
241
MS0017635
Rank:55
Score:
-34.35
242
MS0012191
Rank:4480
Score:-16.95
243
MS0011819
Rank:95
Score:-32.4 244
MS0020927
Rank:236
Score:-29.52
245
MS0014712
Rank:1640
Score:
-22.35
246
MS0017549
Rank:421
Score:-27.64
247
MS0012122
Rank:1000
Score:
-24.53
248
MS0021172
Rank:700
Score:-25.94
249
MS0013315
Rank:1029
Score:
-24.42
250
MS0012257
Rank:1749
Score:-22.07
251
MS0022098
Rank:1761
Score:
-22.04
252
MS0017462
Rank:307
Score:-28.71
253
MS0012337
Rank:3813
Score:
-17.94
254
MS0013810
Rank:1089
Score:-24.17
255
MS0014434
Rank:1306
Score:
-23.39
256
MS0011749
Rank:3808
Score:-17.95
257
MS0011981
Rank:21
Score:
-37.89
258
MS0018746
Rank:451
Score:-27.43
259
MS0019066
Rank:578
Score:
-26.63
260
MS0018262
Rank:1309
Score:-23.38
261
MS0022062
Rank:2460
Score:
-20.32
262
MS0021435
Rank:5194
Score:-15.94
263
MS0022294
Rank:434
Score:
-27.51
264
MS0015058
Rank:486
Score:-27.26
265
MS0019492
Rank:1722
Score:
-22.12
266
MS0015301
Rank:4554
Score:-16.85
267
MS0016846
Rank:1403
Score:
-23.09
268
MS0019472
Rank:1784
Score:-21.95
269
MS0022215
Rank:28
Score:
-36.93
270
MS0021329
Rank:296
Score:-28.87
271
MS0022805
Rank:1993
Score:
-21.44
272
MS0014484
Rank:1922
Score:-21.62
273
MS0022611
Rank:5189
Score:
-15.94
274
MS0012512
Rank:4101
Score:-17.5
275
MS0012753
Rank:5174
Score:
-15.96
276
MS0012133
Rank:2635
Score:-19.93
277
MS0021898
Rank:602
Score:
-26.48
278
MS0019035
Rank:4118
Score:-17.49
279
MS0015409
Rank:1121
Score:-24.1 280
MS0019831
Rank:1659
Score:-22.29
281
MS0012954
Rank:658
Score:
-26.15
282
MS0013800
Rank:4516
Score:-16.91
283
MS0013835
Rank:5649
Score:
-15.26
284
MS0015549
Rank:746
Score:-25.67
285
MS0021727
Rank:1294
Score:
-23.46
286
MS0021613
Rank:4630
Score:-16.75
287
MS0022314
Rank:3014
Score:
-19.27
288
MS0022165
Rank:1637
Score:-22.36
289
MS0019914
Rank:3481
Score:
-18.47
290
MS0021147
Rank:3281
Score:-18.79
291
MS0017526
Rank:5427
Score:
-15.59
292
MS0019932
Rank:3016
Score:-19.27
293
MS0020991
Rank:4139
Score:
-17.45
294
MS0015473
Rank:5303
Score:-15.78
295
MS0021777
Rank:979
Score:
-24.61
296
MS0015519
Rank:228
Score:-29.65
297
MS0019793
Rank:861
Score:
-25.16
298
MS0021746
Rank:391
Score:-27.81
299
MS0022191
Rank:2289
Score:
-20.71
300
MS0022503
Rank:366
Score:-28.1
301
MS0018337
Rank:845
Score:
-25.23
302
MS0016528
Rank:2223
Score:-20.83
303
MS0016876
Rank:1906
Score:
-21.66
304
MS0022738
Rank:3153
Score:-19.03
305
MS0017295
Rank:2357
Score:
-20.56
306
MS0013105
Rank:2559
Score:-20.08
307
MS0021375
Rank:3857
Score:
-17.87
308
MS0018767
Rank:116
Score:-31.91
309
MS0022563
Rank:2550
Score:-20.1 310
MS0019190
Rank:914
Score:-24.93
311
MS0015962
Rank:1372
Score:
-23.19
312
MS0015122
Rank:3505
Score:-18.43
313
MS0015738
Rank:961
Score:
-24.69
314
MS0019292
Rank:3756
Score:-18.03
315
MS0017759
Rank:1187
Score:
-23.86
316
MS0018495
Rank:2078
Score:-21.19
317
MS0017571
Rank:2633
Score:
-19.94
318
MS0011912
Rank:2212
Score:-20.85
319
MS0012518
Rank:3353
Score:
-18.67
320
MS0020524
Rank:533
Score:-26.93
321
MS0021169
Rank:5835
Score:
-15.01
322
MS0012236
Rank:2311
Score:-20.65
323
MS0015964
Rank:5725
Score:
-15.17
324
MS0018001
Rank:827
Score:-25.29
325
MS0022031
Rank:3243
Score:
-18.85
326
MS0013537
Rank:2924
Score:-19.4
327
MS0021954
Rank:3040
Score:
-19.22
328
MS0016623
Rank:4781
Score:-16.54
329
MS0018003
Rank:1921
Score:
-21.62
330
MS0019290
Rank:3679
Score:-18.12
331
MS0021998
Rank:1502
Score:
-22.76
332
MS0019141
Rank:5722
Score:-15.17
333
MS0019143
Rank:233
Score:
-29.53
334
MS0019040
Rank:371
Score:-28.07
335
MS0021388
Rank:751
Score:
-25.64
336
MS0015705
Rank:4220
Score:-17.33
337
MS0018803
Rank:1879
Score:
-21.72
338
MS0011844
Rank:2986
Score:-19.31
339
MS0016642
Rank:3035
Score:
-19.23
340
MS0022752
Rank:3341
Score:-18.69
341
MS0022342
Rank:2074
Score:
-21.21
342
MS0021570_1
Rank:2197
Score:-20.88
343
MS0022678
Rank:192
Score:
-30.34
344
MS0015673
Rank:713
Score:-25.85
345
MS0020536
Rank:1619
Score:
-22.44
346
MS0020370
Rank:2664
Score:-19.88
347
MS0013312
Rank:555
Score:
-26.78
348
MS0018881
Rank:3518
Score:-18.4
349
MS0019477
Rank:2863
Score:
-19.51
350
MS0019065
Rank:5809
Score:-15.04
351
MS0017718
Rank:2397
Score:
-20.47
352
MS0017668
Rank:5814
Score:-15.03
353
MS0016600
Rank:357
Score:
-28.17
354
MS0016673
Rank:2069
Score:-21.22
355
MS0021543
Rank:1
Score:
-44.29
356
MS0021091
Rank:327
Score:-28.52
357
MS0016659
Rank:364
Score:
-28.11
358
MS0013378
Rank:1850
Score:-21.8
359
MS0019564
Rank:2008
Score:-21.4 360
MS0022323
Rank:131
Score:-31.61
361
MS0022196
Rank:405
Score:
-27.74
362
MS0019847
Rank:4842
Score:-16.45
363
MS0021395
Rank:36
Score:
-35.98
364
MS0017869
Rank:3698
Score:-18.1
365
MS0022425
Rank:1161
Score:
-23.93
366
MS0013831
Rank:1034
Score:-24.41
367
MS0020888
Rank:2322
Score:
-20.63
368
MS0021463
Rank:4459
Score:-16.98
369
MS0016833
Rank:1790
Score:
-21.93
370
MS0019735
Rank:3010
Score:-19.28
371
MS0020921
Rank:5403
Score:
-15.63
372
MS0013354
Rank:1404
Score:-23.09
373
MS0014280
Rank:2249
Score:
-20.78
374
MS0021414
Rank:1907
Score:-21.66
375
MS0021470
Rank:11
Score:
-38.94
376
MS0021274
Rank:403
Score:-27.74
377
MS0015458
Rank:22
Score:
-37.58
378
MS0015746
Rank:588
Score:-26.55
379
MS0018108
Rank:4185
Score:
-17.39
380
MS0017367
Rank:263
Score:-29.2
381
MS0014681
Rank:547
Score:
-26.83
382
MS0018351
Rank:2260
Score:-20.77
383
MS0017190
Rank:3137
Score:
-19.05
384
MS0015744
Rank:629
Score:-26.35
385
MS0014323
Rank:640
Score:
-26.26
386
MS0021522
Rank:1990
Score:-21.45
387
MS0021801
Rank:4882
Score:
-16.38
388
MS0019208
Rank:3256
Score:-18.84
389
MS0018016
Rank:2017
Score:
-21.38
390
MS0020177
Rank:4157
Score:-17.43
391
MS0013797
Rank:5168
Score:
-15.97
392
MS0012587
Rank:1135
Score:-24.06
393
MS0014893
Rank:2555
Score:
-20.09
394
MS0016487
Rank:5112
Score:-16.03
395
MS0020294
Rank:2124
Score:
-21.06
396
MS0015934
Rank:5336
Score:-15.73
397
MS0015169
Rank:1397
Score:-23.1 398
MS0012116
Rank:4962
Score:-16.24
399
MS0021642
Rank:2566
Score:
-20.06
400
MS0022529
Rank:3527
Score:-18.39
401
MS0022663
Rank:2666
Score:
-19.88
402
MS0012202
Rank:142
Score:-31.24
403
MS0021694
Rank:1273
Score:
-23.52
404
MS0021606
Rank:3028
Score:-19.25
405
MS0016871
Rank:3166
Score:
-18.99
406
MS0018626
Rank:852
Score:-25.2
407
MS0012397
Rank:1080
Score:
-24.21
408
MS0021137
Rank:2203
Score:-20.87
409
MS0022209
Rank:1732
Score:-22.1 410
MS0021568
Rank:1915
Score:-21.64
411
MS0020665
Rank:5454
Score:
-15.54
412
MS0015418
Rank:5573
Score:-15.37
413
MS0016183
Rank:5501
Score:
-15.46
414
MS0021398
Rank:4408
Score:-17.05
415
MS0016203
Rank:189
Score:
-30.43
416
MS0012509
Rank:4565
Score:-16.83
417
MS0016512
Rank:3607
Score:
-18.26
418
MS0018800
Rank:703
Score:-25.92
419
MS0012712
Rank:3027
Score:
-19.25
420
MS0012109
Rank:2523
Score:-20.16
421
MS0017621
Rank:3742
Score:
-18.04
422
MS0020110
Rank:1988
Score:-21.45
423
MS0011518
Rank:4043
Score:
-17.59
424
MS0011766
Rank:5733
Score:-15.16
425
MS0011711
Rank:24
Score:
-37.49
426
MS0011996
Rank:93
Score:-32.48
427
MS0021138
Rank:545
Score:
-26.84
428
MS0014488
Rank:1873
Score:-21.73
429
MS0012102
Rank:439
Score:
-27.47
430
MS0011541
Rank:2252
Score:-20.78
431
MS0012600
Rank:344
Score:
-28.32
432
MS0014542
Rank:12
Score:-38.68
433
MS0012333
Rank:187
Score:
-30.48
434
MS0015329
Rank:317
Score:-28.6
435
MS0022769
Rank:2
Score:
-43.58
436
MS0012680
Rank:581
Score:-26.61
437
MS0015622
Rank:2936
Score:-19.4 438
MS0012162
Rank:636
Score:-26.32
439
MS0011787
Rank:1204
Score:
-23.76
440
MS0016716
Rank:1140
Score:-24.03
441
MS0014543
Rank:3
Score:
-42.27
442
MS0019880
Rank:755
Score:-25.63
443
MS0018725
Rank:77
Score:
-32.92
444
MS0022360
Rank:149
Score:-31.02
445
MS0016263
Rank:2641
Score:
-19.92
446
MS0015373
Rank:1443
Score:-22.95
447
MS0014034
Rank:1494
Score:
-22.78
448
MS0020769
Rank:3571
Score:-18.33
449
MS0021508
Rank:691
Score:
-25.98
450
MS0012703
Rank:2108
Score:-21.09
451
MS0020359
Rank:4674
Score:-16.7 452
MS0014729
Rank:3664
Score:-18.15
453
MS0022560
Rank:791
Score:
-25.49
454
MS0011911
Rank:1954
Score:-21.56
455
MS0018005
Rank:5486
Score:
-15.47
456
MS0012169
Rank:1772
Score:-22
457
MS0011831
Rank:1384
Score:
-23.13
458
MS0021161
Rank:81
Score:-32.84
459
MS0021956
Rank:281
Score:-29 460
MS0014872
Rank:186
Score:-30.48
461
MS0021246
Rank:4181
Score:-17.4 462
MS0021770
Rank:474
Score:-27.31
463
MS0016542
Rank:1352
Score:
-23.23
464
MS0012776
Rank:242
Score:-29.45
465
MS0015051
Rank:2312
Score:
-20.65
466
MS0019196
Rank:125
Score:-31.74
467
MS0018564
Rank:310
Score:
-28.67
468
MS0018145
Rank:4313
Score:-17.18
469
MS0021108
Rank:1972
Score:
-21.51
470
MS0014646
Rank:3241
Score:-18.86
471
MS0012437
Rank:5053
Score:
-16.11
472
MS0016592
Rank:4780
Score:-16.54
473
MS0022230
Rank:1442
Score:
-22.95
474
MS0021799
Rank:1695
Score:-22.18
475
MS0012264
Rank:2105
Score:
-21.11
476
MS0016619
Rank:1984
Score:-21.47
477
MS0017254
Rank:3726
Score:
-18.06
478
MS0020461
Rank:2892
Score:-19.46
479
MS0012630
Rank:3252
Score:
-18.84
480
MS0014756
Rank:58
Score:-34.08
481
MS0011672
Rank:1067
Score:
-24.29
482
MS0012728
Rank:1166
Score:-23.92
483
MS0011645
Rank:4820
Score:
-16.48
484
MS0011653
Rank:2839
Score:-19.54
485
MS0012641
Rank:345
Score:
-28.32
486
MS0019147
Rank:1573
Score:-22.57
487
MS0019122
Rank:2101
Score:
-21.12
488
MS0014761
Rank:4058
Score:-17.57
489
MS0019897
Rank:3294
Score:
-18.77
490
MS0014674
Rank:4212
Score:-17.34
491
MS0014402
Rank:5440
Score:
-15.57
492
MS0021126
Rank:1711
Score:-22.15
493
MS0016367
Rank:5087
Score:
-16.06
494
MS0015773
Rank:4453
Score:-16.99
495
MS0016519
Rank:4759
Score:
-16.58
496
MS0012741
Rank:1755
Score:-22.05
497
MS0011793
Rank:2553
Score:
-20.09
498
MS0012583
Rank:2987
Score:-19.31
499
MS0011416
Rank:2906
Score:
-19.44
500
MS0014376
Rank:3586
Score:-18.3
501
MS0014325
Rank:3993
Score:
-17.66
502
MS0019243
Rank:4930
Score:-16.3
Table S2. Displacement of radiolabeled b-LEN by a compound with chemical similarity to MS0015203.
MS0016574 is a structurally related compound to MS0015203, both of which are in cluster 8. Rat
hypothalamic membranes were incubated with 3 nM [125I]b-LEN in the presence or absence of the
indicated compounds (0-10 M), percent displacement of bound b-LEN and IC50 were calculated. Data
are mean ± SE of 2 independent experiments performed in triplicate.