1. Solid-State NMR Utility in API and Formulation Process Development Robert Wenslow VP Business Development Crystal Pharmatech www.crystalpharmatech.com

2. 2 Biopharmaceutics Organic Process Granulation Bulk API Characterization In Formulation Samples Tableting In vivo performance Excipient Interactions Produce Stable, Single Phase Bulk Engineering Processing Conditions Solvent Stystem Milling Issues Compaction Analytical Phys/Chem Stability Spec. Justification Areas of Application

3. 3  Salt Disproportionation  Polymorphs  Solvates and Hydrates  Amorphous dispersions  Motivation is regulation Challenging Pharmaceutical Issues

4. 4 Solid-State NMR http://www.dur.ac.uk/resources/SSNMR/Training_course_PH.pdf Molecular Tumbling Rigid Solid Orientation leads to CS difference

5. 5 Cross-Polarization (CP/MAS) J. Chem. Phys. 1973, 59, 569 1H1H 13 C Detect Decouple (90) Contact Time Signal Intensity Contact Time Signal Buildup through dipole coupling (D) 1H1H 13 C D µ 1/r 6

6. ** K-salt disproportionates in water to the free acid 19 F NMR was used to determine the kinetics Fast experiments, quantitative information Can also probe in formulation Structure: Disproportionantion 6

7. Lower decoupling obscures N’s connected to H Disappearance of 2 peak predicted Salt forms at 3 Nitrogen Contact time can also be used to discriminate 4 3 Useful information but long measuring times 7 Structure: Salt Formation

8. 19 F NMR spectra of I and II 19 F relaxation curves for I and II At 8.5 seconds FII shows zero signal 19 F relaxometry can be used 8 Polymorph Quantitation

9. CP discriminates against the more mobile regions DP discriminates against the more rigid regions Spectral editing combination is powerful to study solvates * Peaks from EtOH Split –CH 3 indicates multiple environments Straight forward measurements High information content Form II CP/MAS Form II DP/MAS 9 Solvate Identification

10. 19 F SSNMR was used, measurements done at 5 o C Formulation: API + PEG 600 At 40 mgs/mL API completely dissolved At 80 and 100 mgs/mL shoulder observed L454 Freebase 10 API in Drug Product

11. Expanded spectrum Shoulder at 80 and 100 mgs/mL due to crystalline freebase. Rapid measurements, quantitative estimation of solubility possible. 11 API in Drug Product

12. 1 H NMR was used Rigid: Gaussian Mobile: Lorentzian Fitting provides quantitation Amorphous content 22.5% Extremely rapid measurements, quantitative No chemical shift resolution 12 Amorphous API

13. Amorphous has a very short T 1 ~ 250 ms Crystal had a very long T 1 ~ 25 sec 31 P NMR Material stuck on pins ~ 12 wt% amorphous 10X compaction at 200 MPa (RT) ~ 5 wt% 10X compaction at 200 MPa (85 o C) ~ 2 wt% 13 Amorphous API

14. Broadening due to defects or phase separated amorphous?? -130-120-110-100-90 Time = 0 2 weeks ambient 2 weeks 25C/60%RH 2 weeks 40C/75%RH 14 Amorphous API

15. M(tau)/M0= 1-2*exp(-tau/T1) -130-120-110-100-90 Pure Crystalline T1 = 5 seconds Pure Amorphous T1 = 1 second Each Phase in multi-phase system will yield unique T1 value 15 Amorphous API

16. -130-120-110-100-90 End of Drying Sample (MAS) T1 Filter 2.55 sec Can detect amorphous content without any apriori knowledge of system. Was also used identify presence of multiple crystalline phases Monitoring in-process samples 16 Amorphous API

17. -130-120-110-100-90 Time = 0 2 weeks ambient 2 weeks 25C/60%RH 2 weeks 40C/75%RH Intensity directly proportional to amorphous content Qualitative amorphous content readily achieved LOD exceedingly low (limited only by NMR time) Quantitation requires calibration curve Monitoring stability samples 17 Amorphous API

18. 18 DECRA

19. 19 19 F CP/MAS DECRA Component 2 200msec T1rho 65% of total spectra Component 1 39msec T1rho 35% of total spectra -100-80-60-40 -20 Stability Sample Previous ID of multiple Phases by 1 H T 1rho Filter 1 H T 1rho DECRA

20. 20 Can we quantify second phase  API process involves desolvation to get the anhydrous form  Material forms amorphous on compaction  A second phase observed in 19 F SSNMR spectra for different batches  Similar XRPD and DSC 19 F CP/MASXRPD DECRA Polymorph ID

21. 21 -CF 3 F  1 H T 1  DECRA 19 F CP/MAS Component 2 6msec T1rho 20% of total spectra Component 1 20msec T1rho 80% of total spectra DECRA

22. 22 Wet milling in IPAc followed by Drying at 50 o C recommended DECRA Driving Process Definition

23. 23 1 H T 1  DECRA Through 19 F 1 H T 1  DECRA 13 C CP/MAS Component 2 91msec T1rho 22% of total spectra Component 1 193msec T1rho 78% of total spectra T 1rho Wt% Comp 113370 Comp 25530 DECRA My API doesn’t have a 19 F

24. Heteronuclear Dipolar Correlation  1 H- 13 C, 1 H- 15 N, and 1 H- 23 Na HETCOR spectra for a hydrated API Cryst. Growth & Des., 2006, 6, 2333-2354. Correlations indicate atoms near in space (~3 Å ) 13 C 15 N 23 Na 1H1H 24

25. 25 Amorphous Dispersions

26. 2D 1 H- 19 F Correlation  1 H- 19 F CP-HETCOR easily proves molecular association on the < 10 Å scale  Experiments such as these take 1-2 hours to perform for typical drug loads (20-60% w/w) 500  s 2 ms (spin diffusion) Mol. Pharmaceutics, 7, 1667–1691 (2010). n N O OH OHO FF PVP Solid amorphous solution Diflunisal

27. Amorphous Dispersions  A dispersion that greatly improves the dissolution of tenoxicam in water (via a high degree of supersaturation)  Contains four discrete components Polyvinylpyrrolidine Solid amorphous solution = tenoxicam (singly ionized) = L-arginine (singly ionized) = L-arginine (zwitterionized) J. Pharm. Sci. 2012, 101, 641-663.

28. Nanocrystallline dispersion Pharm. Res. 2012, 29, 1866-1881 Polymer Crystalline Domains ~50 nm

29. 29 Concluding Thoughts  Multitude of options to characterize API and drug product material  Relaxation methodology very powerful  Expanding into 2D offers significant structure information