Double effect molecular switch leads to a novel potent negative allosteric modulator of metabotropic glutamate receptor 5

X. Gómez-Santacanaa,b,c X. Rovirab,d,e, J. Daltonb, C. Goudetd,e, J. P. Pin d,e,, P. Gorostiza c,f,g, J. Giraldob and A. Llebaria*a  

a Laboratory of Medicinal Chemistry, Institute for Advanced Chemistry of Catalonia (IQAC-CSIC), 08034 Barcelona, Spain. E-mail: amadeu.llebaria@cid.csic.es b Laboratory of Molecular Neuropharmacology and Bioinformatics, Institut de Neurociències and Unitat de Bioestadística, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain. c Institute for Bioengineering of Catalonia (IBEC), 08028 Barcelona, Spain. d Institut de Génomique Fonctionnelle, CNRS, UMR-5203, Université de Montpellier, F-34000 Montpellier, France. e INSERM, U661, F-34000 Montpellier, France. f Network Biomedical Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN) g Catalan Institution for Research and Advanced Studies (ICREA), Barcelona, Spain.

Supplementary Information

1. Nuclear Magnetic Resonance (NMR)

2. High Performance Liquid Chromatography Coupled to Photodiode Array (HPLC/PDA)

3. Liquid Chromatography Coupled to Mass Spectrum (LC/MS)

4. Elemental Composition by High Resolution Mass Spectroscopy (HRMS)

Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications.This journal is © The Royal Society of Chemistry 2014

1. Nuclear Magnetic Resonance (NMR) spectra

1.1. Compound 16

1.1.1. 1H-NMR (400 MHz, CDCl3)

1.1.2. 13C-NMR (101 MHz, CDCl3)

1.2. Compound 17

1.2.1. 1H-NMR (400 MHz, CDCl3)

1.2.2. 13C-NMR (101 MHz, CDCl3)

1.3. Compound 20

1.3.1. 1H-NMR (400 MHz, CDCl3)

1.3.2. 13C-NMR (101 MHz, CDCl3)

1.4. Compound 21

1.4.1. 1H-NMR (400 MHz, CDCl3)

1.4. Compound 24

1.4.1. 1H-NMR (400 MHz, CDCl3)

1.5. Compound 25

1.5.1. 1H-NMR (400 MHz, CDCl3)

2. High Performance Liquid Chromatography Coupled to Photodiode Array (HPLC/PDA)

2.1. Compound 16

Multi Sample Summary Project Name: mGluReported by User: System

Page: 1 of 1Report Method: Multi Sample Summary Printed 3:01:56 PM 3/6/2014

S A M P L E I N F O R M A T I O N

Acquired By: System Sample Name: Compund 16Date Acquired: 12/19/2013 7:41:54 PM Sample Type: Unknown Acq. Method Set: XGS RC_MT 1 Vial: 58 Date Processed: 3/6/2014 3:00:45 PM Injection #: 1 Processing Method: xgs_254 Injection Volume: 5.00 ulChannel Name: Wvln Ch1 Run Time: 12.0 MinutesProc. Chnl. Descr.: PDA 254.0 nm Sample Set Name: XGS

AU

0.00

0.10

0.20

0.30

0.40

0.50

0.60

Minutes1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00

3.36

1

4.18

24.

420

4.92

9

1

2

3

4

RT Area % Area Height

3.361

4.182

4.420

4.929

26425

3571148

98646

47209

0.71

95.40

2.64

1.26

6893

611254

19064

8423

2.2. Compound 17

Multi Sample Summary Project Name: mGluReported by User: System

Page: 1 of 1 Report Method: Multi Sample Summary Printed 3:07:35 PM 3/6/2014

S A M P L E I N F O R M A T I O N

Acquired By: System Sample Name: Compund 17 Date Acquired: 12/19/2013 2:59:20 PM Sample Type: Unknown Acq. Method Set: XGS RC_MT 1 Vial: 36 Date Processed: 3/6/2014 3:06:45 PM Injection #: 1 Processing Method: xgs_254 Injection Volume: 5.00 ulChannel Name: Wvln Ch1 Run Time: 12.0 MinutesProc. Chnl. Descr.: PDA 254.0 nm Sample Set Name: XGS

AU

0.00

0.20

0.40

0.60

0.80

1.00

1.20

1.40

1.60

Minutes1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00

3.26

5

3.84

7

1

2

RT Area % Area Height

3.265

3.847

6336952

21508

99.66

0.34

1579560

6304

3. Liquid Chromatography coupled to Mass Spectrum (HPLC/MS)

3.1. Compound 16

3.1.1. UV-Vis absorbance chromatogram and TIC chromatogram

Time0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00

%

-18

82

0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00

AU

0.0

5.0e-2

1.0e-1

1.5e-1

2.0e-1

Compound 16 5: Diode Array Range: 2.865e-1

Area9.08

98.0%31154

Compound 16 1: TOF MS ES+ TIC

2.22e59.11

0.958.67

9.60

12.2010.50

11.48

Time0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00

%

-16

84

0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00

AU

0.0

5.0e-2

1.0e-1

1.5e-1

2.0e-1

Blank XGS 5: Diode Array Range: 3.676e-2

0.68 1.50 12.00

Blank XGS 1: TOF MS ES+ TIC

2.50e5

9.620.95 8.68

12.21

Compound 16

RT Height Area % Area

9.083 271822 31153.928 97.5%

9.317 3284 247.251 0.8%

9.583 5605 548.625 1.7%

3.1.2. TOF Mass Spectrum

mGlu

m/z100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600 1700

%

0

100Compound 16 185 (9.158) 1: TOF MS ES+

3.49e4363

359279208143124

725

365

366

391 404 706661573454

!509 601

727

728

729

730

1483731 1089778 822!

1183 1505

!1588 !

1744

3.1.3. Elemental Composition Analysis

Elemental Composition Report Page 1

Single Mass Analysis Tolerance = 10.0 PPM / DBE: min = -1.5, max = 50.0Selected filters: None

Monoisotopic Mass, Even Electron Ions111 formula(e) evaluated with 1 results within limits (up to 50 closest results for each mass)Elements Used:C: 10-25 H: 15-20 N: 0-5 O: 0-5 Cl: 0-5

m/z100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600 1700

%

0

100

mGluCompound 16 185 (9.158) 1: TOF MS ES+

3.49e4363

359279208143124

725365

366426 706661573555 728 14831089778 866 1183 1505 1588 1744

Minimum: -1.5Maximum: 5.0 10.0 50.0

Mass Calc. Mass mDa PPM DBE i-FIT Formula

363.0897 363.0900 -0.3 -0.8 14.5 76.1 C21 H16 N2 O2 Cl

3.1. Compound 17

3.1.1. UV-Vis absorbance chromatogram and TIC chromatogram

Time0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00

%

-8

92

0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00

AU

0.0

5.0e-2

1.0e-1

1.5e-1

2.0e-1

2.5e-1

3.0e-1

Compound 17 5: Diode Array Range: 4.144e-1

Area8.02

99.5%63817

Compound 17 1: TOF MS ES+ TIC

5.02e58.01

0.9512.219.57 11.49

Time0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00

%

-16

84

0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00

AU

0.0

5.0e-2

1.0e-1

1.5e-1

2.0e-1

Blank XGS 5: Diode Array Range: 3.676e-2

0.68 1.50 12.00

Blank XGS 1: TOF MS ES+ TIC

2.50e5

9.620.95 8.68

12.21

Compound 17

RT Height Area % Area

8.017 411373 63816.559 100%

3.1.2. TOF Mass Spectrum

mGlu

m/z100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600 1700

%

0

100Compound 17 163 (8.097) 1: TOF MS ES+

1.06e5363

238208143

!90

359245

365

366725

404426 475691591

!539

!657

7281111

!837787 937 1151 1471

3.1.3. Elemental Composition Analysis

Elemental Composition Report Page 1

Single Mass Analysis Tolerance = 10.0 PPM / DBE: min = -1.5, max = 50.0Selected filters: None

Monoisotopic Mass, Even Electron Ions111 formula(e) evaluated with 1 results within limits (up to 50 closest results for each mass)Elements Used:C: 10-25 H: 15-20 N: 0-5 O: 0-5 Cl: 0-5

m/z100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600 1700 1800

%

0

100

mGluCompound 17 163 (8.097) 1: TOF MS ES+

1.06e5363

23820814390 359327

365

725404 426 475 691591 747 1111837 937 1151 1471

Minimum: -1.5Maximum: 5.0 10.0 50.0

Mass Calc. Mass mDa PPM DBE i-FIT Formula

363.0906 363.0900 0.6 1.7 14.5 61.0 C21 H16 N2 O2 Cl

4. Elemental Composition by High Resolution Mass Spectroscopy (HRMS)

4.1. Compound 21

Elemental Composition Report Page 1

Single Mass Analysis Tolerance = 10.0 PPM / DBE: min = -1.5, max = 50.0Selected filters: None

Monoisotopic Mass, Even Electron Ions28 formula(e) evaluated with 1 results within limits (all results (up to 1000) for each mass)Elements Used:C: 10-20 H: 10-20 N: 0-5 O: 0-5 Cl: 1-1

m/z100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600

%

0

100

mGluCompound 21 21 (1.033) 1: TOF MS ES+

54.7258

21479 179 359303 515394431 850541 645667 703 773 1073867908 981 1150 1308 1339 1453 1566

Minimum: -1.5Maximum: 5.0 10.0 50.0

Mass Calc. Mass mDa PPM DBE i-FIT Formula

258.0685 258.0686 -0.1 -0.4 9.5 14.0 C15 H13 N O Cl

4.2. Compound 24

Elemental Composition Report Page 1

Single Mass Analysis Tolerance = 10.0 PPM / DBE: min = -1.5, max = 50.0Selected filters: None

Monoisotopic Mass, Even Electron Ions14 formula(e) evaluated with 1 results within limits (all results (up to 1000) for each mass)Elements Used:C: 10-15 H: 10-15 N: 0-3 O: 0-3 Cl: 0-1

m/z100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600 1700

%

0

100

mGluCompound 24 15 (0.721) 1: TOF MS ES+

1.12e3255

79 20815780256 305 391 509432 645554 702 776 850 908 925 1047 1129

Minimum: -1.5Maximum: 5.0 10.0 50.0

Mass Calc. Mass mDa PPM DBE i-FIT Formula

255.0769 255.0770 -0.1 -0.4 10.5 1.2 C14 H11 N2 O3

4.3. Compound 25

Elemental Composition Report Page 1

Single Mass Analysis Tolerance = 10.0 PPM / DBE: min = -1.5, max = 50.0Selected filters: None

Monoisotopic Mass, Even Electron Ions15 formula(e) evaluated with 1 results within limits (all results (up to 1000) for each mass)Elements Used:C: 10-15 H: 10-15 N: 0-3 O: 0-3 Cl: 0-1

m/z100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600 1700

%

0

100

mGluCompound 25 16 (0.772) 1: TOF MS ES+

813225

21079 124691476432388226 349 520 673564 693 850776 888 927 1129983 12051222 1369 1388

Minimum: -1.5Maximum: 5.0 10.0 50.0

Mass Calc. Mass mDa PPM DBE i-FIT Formula

225.1027 225.1028 -0.1 -0.4 9.5 8.8 C14 H13 N2 O