High-resolution probes for 19 F applications. Aitor Moreno Bruker BioSpin, Switzerland BeNeLux NMR User Meeting 2016, Brussels 25 th of November PDF Free Download

High-resolution probes for F applications Aitor Moreno Bruker BioSpin, Switzerland BeNeLux NMR User Meeting 206, Brussels 25 th of November 206 Fluorite (CaF 2 ), lat. fluores Innovation with Integrity

2 Why F NMR? I=/2, 00% natural abundance γ F = γ H 94.09% Large chemical shift range (+440 to -450 ppm) High sensitivity of F on chemical environment Reference substance: CCl 3 F (δ F =0ppm) Use of F in drugs: Higher metabolic stability Increased membrane permeability Tune affinity to target enzyme F containing insecticides / herbicides / fungicides F containing synthetic polymers

F NMR applications Probe design (inner / outer coil) Double resonance applications BBO H&F BBFO SmartProbe Triple resonance applications TXO probe TBO probe CryoProbes for highest F sensitivity TCI H&F cryoprobe QCIF cryoprobe 3

4 RF design & RF efficiency: B / I coil S N a U I U N a w. M o. V. h. (B / I coil) 4. k. Df. R. T How can the RF efficiency be improved? Optimize RF coil design & RF coil materials Reduce RF coil diameter

Probehead Type Classification Proton inner coil: INVERSE X-nucleus inner coil: OBSERVE H (Proton) X-nucleus broad-band { 09 Ag 3 P} Double resonance broad-band inverse BBI Double resonance broad-band observe BBO 5

Probehead Type Classification Proton inner coil: INVERSE X-nucleus inner coil: OBSERVE H (Proton) X-nucleus broad-band X-nucleus broad-band H (Proton) Double resonance broad-band inverse BBI Double resonance broad-band observe BBO 6

H sensitivity of BBI and BBO BBI (Proton inner coil) H performance: sensitivity and resolution: BBO (Proton outer coil) BBI BBO 8.8 8.6 8.4 8.2 8.0 7.8 7.6 7.4 7.2 7.0 ppm 8.8 8.6 8.4 8.2 8.0 7.8 7.6 7.4 7.2 7.0 ppm H spectra, Cyclosporin 7

3 C sensitivity of BBI and BBO 50mM cyclosporin A BBI ( 3 C outer coil) 50mM cyclosporin A 80 70 60 50 40 30 20 0 00 90 80 70 60 50 40 30 20 ppm Current Data Parameters NAME BBI_Performance EXPNO 4 PROCNO F2 - Acquisition Parameters Date_ 20040629 Time 3. INSTRUM av400 PROBHD 5 mm BBI H-BB PULPROG deptsp35 TD 65536 SOLVENT C6D6 NS 256 DS 2 SWH 8382.354 Hz FIDRES 0.280492 Hz AQ.7826564 sec RG 6384 DW 27.200 usec DE 6.00 usec TE 300.5 K CNST2 35.0000000 D 2.00000000 sec d2 0.00370370 sec d2 0.00002000 sec DELTA 0.0000553 sec MCREST 0.00000000 sec MCWRK 0.0500000 sec ======== CHANNEL f ======== NUC 3C P 2.20 usec P2 2000.00 usec PL0 20.00 db PL -6.00 db SFO 00.62208 MHz SP2 0.43 db SPNAM2 Crp60comp.4 SPOFF2 0.00 Hz ======== CHANNEL f2 ======== CPDPRG2 waltz6 NUC2 H P3 0.50 usec p4 2.00 usec PCPD2 80.00 usec PL2 -.00 db PL2 6.64 db SFO2 400.36005 MHz F2 - Processing parameters SI 3072 SF 00.627690 MHz WDW EM SSB 0 LB.00 Hz GB 0 PC.40 BBO ( 3 C inner coil) 3 C DEPT35, Cyclosporin 80 70 60 50 40 30 20 0 00 90 80 70 60 50 40 30 8 20

0 BBO design Extension to F BBO H&F BBFO SmartProbe BB { 09 Ag 3 P} BBF { 09 Ag 3 P, F} H / F H H or F

BBO H&F Tuned to H BBO H&F H NMR 2 J H,F BB { 09 Ag 3 P} F H O F 3 C F F 2 J H,F H / F Tuned to H,,2,3,3,3-hexafluoropropyl-ethyl-ether in CDCl 3

BBO H&F Tuned to F BBO H&F F NMR 2 J H,F BB { 09 Ag 3 P} 2 J H,F F H O F 3 C F F H / F Tuned to F,,2,3,3,3-hexafluoropropyl-ethyl-ether in CDCl 3 2

BBFO SmartProbe BBFO SmartProbe H NMR BBF: F H 2 J H,F H H{ F} BBF: tuned to F 3

4 BBFO SmartProbe D F NMR with H BB decoupling F 3700: 8060: F{ H} F H O F 3 C F F

F sensitivity - with and without H decoupling TFT Hexafluoropropylethyl-ether Fluoro-uracil F H O F 3 C F F F F{ H} SNR ratio (decoupled/coupled) 2.7 2.2 5.6 400MHz, lb=0.5hz, ppm noise range 5

BBFO SmartProbe F - H correlations 2D HOESY BF 4 - BF 4 - In CD 2 Cl 2 6

7 SmartProbe F background signal SmartProbe allround probe for many different applications With F background reduction Standard measures Probe

Triple resonance applications simultaneous BB decoupling on two nuclei Double Resonance: Triple Resonance: F{ H}, H{ F} 3 C{ H, F}, F{ H, 3 C}, H{ F, 3 C} F 3 C H BB decoupling H BB decoupling F BB decoupling SmartProbe 2 RF channels Proton amplifier (QNP switch) TXO or TBO probe 3 RF channels 2 Proton amplifiers

Triple resonance F applications TXO probes ( 3 C, F, H) F{ H}, H{ F} and 3 C{ H, F}, F{ H, 3 C}, H{ F, 3 C} TXO probes avialable in two «flavours»: 3 C Priority on F (F/C/H) Priority on 3 C (C/F/H) VT range: -50 to +50 C F background reduction measures Full automation for all experiments with C, F & H H & F Priority on 3 C: TXO C/F/H 20

400MHz TXO probe Priority on F F sensitivity with H decoupling SiNo: 827: (lb=0.5 Hz; ppm noise region) F Sensitivity: 0.05% Trifluorotoluene in CDCl 3 2

22 400MHz TXO probe Priority on F D H NMR F H O F 3 C F F

23 400MHz TXO probe Priority on F D H NMR with F and 3 C decoupling H H{ F} *Impurity H{ F, 3 C} * * * *

24 400MHz TXO probe Priority on F D F NMR with H BB decoupling F F{ H} F H O F 3 C F F

25 400MHz TXO probe Priority on F D F NMR with H BB decoupling F F{ H} F H O F 3 C F F

26 400MHz TXO probe Priority on F D F NMR with H and 3 C BB decoupling F F{ 3 C} F H O F 3 C F F F{ 3 C, H}

27 400MHz TXO probe Priority on F H, 3 C HMBC F H O F 3 C F F

28 400MHz TXO probe Priority on F H, 3 C HMBC with F decoupling in F F H O F 3 C F F

30 400MHz TXO probe Priority on 3 C D 3 C NMR F priority (F/C/H) with H and F BB decoupling 3 C priority (C/F/H)

3 5mm TBO BB-F/H Probe Optimized sensitivity for X nuclei detection Triple resonance: X-F-H with X broadband BB { 5 N 3 P} BB broadband ( 3 P- 5 N) Full automation for all experiments with X, F & H H & F TBO BB-F/H

32 5mm TBO BB-F/H Probe F, 3 C HSQC 0 mg fluorouracil in 0.5 ml DMSO-d 6 (with H BB decoupling) ns = 2k x 28 data points 2.5 min. experimental time

34 CryoProbe: RF coil temperature S/N versus temperature (K) 4 S N a U I U N a w. M o. V. h. (B /I coil) 4. k. Df. R. T 3 2 25K 293K CryoProbe (He, ~20K) CryoProbe Prodigy (N 2, ~85K)

Helium H&F CryoProbes TCI H&F CryoProbes with H coil tuneable to F Probes with highest F sensitivity (without H decoupling) H / F F{ H} and H{ F} experiments not possible Good for F if no alternative available 3 C & 5 N Suitable for F with broad lines (PrOF NMR) PrOF NMR: Protein Observed Fluor NMR TCI H&F 35

36 QCI HFCN CryoProbe H/F/C/N probe, fixed frequency for Proton, Fluor, Carbon and Nitrogen Probes with highest F sensitivity (with H decoupling) H & F All F{ H} and H{ F} experiments possible Allow for any combination of nuclei with 4 channel spectrometer 3 C & 5 N Useful for F ligand based screening QCI HFCN

F labeled ligand in protein complex PROTEIN: 8kDa, < mm F LIGAND: 2-fold excess D F{ H} NMR Spin Echo CPMG Free ligand Linewidth = 4Hz Bound ligand Linewidth = 8Hz QCIF CryoProbe 37

38 Summary Probes for F NMR applications Probe Field (MHz) Benefits Applications BBFO Smartprobe QCI (HFCN) Cryoprobe 400 600 F{ H}, H{ F} and X{ H}, H{X} VT Range -50 to +80 C 500 800 F{ H}, H{ F} and full range of HFCN 4 channel experiments Small Molecules and Polymers Proteins, Nucleic Acids, Drug Discovery TXO and TBO RT probes 400 800 F{ H}, H{ F} and X{ H, F}, H{ F,X} VT Range -50 to +80 C H&F BBO 400 700 H Channel tunable to F (no H decoupling)* Small Molecules, Polymers, Proteins, Drug Discovery Quick F experiments no decoupling TCI H&F Cryoprobe 400-000 H Channel tunable to F (no H decoupling)* Proteins, Nucleic Acids with broad F signals Cryoprobe Prodigy 400-700 H Channel tunable to F (no H decoupling)* Proteins, Nucleic Acids with broad F signals * Not available in automation with nanobay consoles

Acknowledgements Application colleagues Bruker Biospin Rainer Kümmerle, Helena Kovacs, Clemens Anklin R&D Bruker BioSpin Thank you!

Innovation with Integrity

High-resolution probes for 19 F applications. Aitor Moreno Bruker BioSpin, Switzerland BeNeLux NMR User Meeting 2016, Brussels 25 th of November 2016