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: lakshmi.devi@mssm.edu

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.