© Pfizer Limited Cap and CASE: a step forward towards automatic structure elucidation Anna Codina, Ruth Boetzel, Ingrid Hamernig and Don Richards Structure Elucidation, ARD, PGRD
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3 Good ResultsMicro + Training
© Pfizer Limited Introduction 2.Applications/Examples: Identification of Impurities Challenging molecules: Arylpyrazoles Regulatory documents generation/checking 3.Conclusions
© Pfizer Limited Structure Elucidation LC - NMR ( 1 H) Structure ~> 0.5% Isolation 2D NMR + MS Report / Database LC-MS Low level impurities identification Proof of Structure / Structure confirmation for regulatory documentation
© Pfizer Limited NMR: How can we work faster? acquisition processing analysis results/report/LNB database
© Pfizer Limited Acquisition - Sensitivity of NMR spectroscopy MSlow nanogram UVlow nanogram NMR microgram
© Pfizer Limited NMR signal to noise
© Pfizer Limited Improving sensitivity High magnetic fields Performance enhancement of RF probe and electronics < V detection coil (microcoils) Cold probes and preamplifiers Coil built from low- loss materials (superconductors) Energy levels & population distribution Hyperpolarisation (DNP)
© Pfizer Limited Microcoils 5 mm Probe CapNMR TM Microcoil
© Pfizer Limited Microcoils and microflow NMR Flow cell
© Pfizer Limited Automation - OMNMR
© Pfizer Limited Analysis: Computer Assisted Structure Elucidator NMR: pp 2D MS: MF Generate MCD Generate Structures From MCD/s Check MCD Find the Best Structure ! Check data/ add information
© Pfizer Limited Introduction 2.Applications/Examples: Identification of Impurities Challenging molecules: Arylpyrazoles Regulatory documents generation/checking 3.Conclusions
© Pfizer Limited Structure elucidation of low level impurities ? LC - NMR ( 1 H) Structure ~> 0.5% Isolation 2D NMR + MS Report / Data base LC-MS
© Pfizer Limited CapNMR data for 1 of the impurities 9 mg/60ul injection ug in the active volume 13 C 27 h HMBC 4 h
© Pfizer Limited full data sets to analyse!
© Pfizer Limited Computer Assisted Structure Elucidation (CASE) Molecular Connectivities Diagram (MCD) GENERATION of STRUCTURES from MCD
© Pfizer Limited /97953 – 6h 52 m THE BEST STRUCTURE d_{N}(^{13}C) d_{N}(^{1}H) d_{A}(^{13}C+^{1}H) 1.570
© Pfizer Limited R R R
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© Pfizer Limited Introduction 2.Applications/Examples: Identification of Impurities Challenging molecules: Arylpyrazoles Regulatory documents generation/checking 3.Conclusions
© Pfizer Limited Arylpyrazoles
© Pfizer Limited CASE of Example mg C 15 H 8 O 1 N 4 F 8 Cl 2 1 H, 13 C, 1 H- 13 C HSQC, 1 H- 13 C/N- HMBC
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© Pfizer Limited No Filter: 3325/3409 – 2 minFilter: C CS and internal bad list 3/3409 – 2 min
© Pfizer Limited CASE of Example 2 – 0.5 mg C 15 H 7 O 2 N 3 F 8 Cl 2 1 H, 13 C, 1 H- 13 C HSQC, 1 H- 13 C/ 15 N- HMBC, 1 H 19 F-HSQC
© Pfizer Limited From MCD 26: 1476 – 11 s ACD + Arylpyrazole fragments
© Pfizer Limited CASE of Arylpyrazoles – challenging molecules Less information More Human intervention
© Pfizer Limited Introduction 2.Applications/Examples: Identification of Impurities Challenging molecules: Arylpyrazoles Regulatory documents generation/checking 3.Conclusions
© Pfizer Limited Cap and CASE in regulatory Regulatory documentation is required for drugs to go through clinical trials Structure confirmation or full prove of structure Not sample limited Cap is ideal: ‘all in one’ ( 13 C) and automation CASE: –Used as a checking tool for H and C assignments: data are peak picked and contain 13 C direct –Can be used as a automatic structure verification and assignment tool and maybe for proof of structure documents
© Pfizer Limited CASE and regulatory data
© Pfizer Limited Conclusions CASE can be used to elucidate the structure of unknowns, such as impurities. Training systems are a powerful tool, especially for molecules with low number of connectivities. Good data required and limited overlapping. Not a easy tool to use. Generation of structures from an unbiased position, avoiding wrong assumptions. The combination of capillary microcoil NMR and CASE significantly decreases the level of human intervention in SE.
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© Pfizer Limited Acknowledgements SE team: Ingrid Hamernig Protasis: Dean Olson, Bob Albrecht, Tim Peck and David Strand ACD labs: Ryan Sasaki
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