a pharmaceutical industry perspective: a new era in...
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Kelly Zhang
Genentech Inc., 1 DNA Way, South San Francisco, CA 94080, USA
Input to NAS Study:A Research Agenda for a New Era in Separations Science
May 7-8, 2018, Irvine, CA
A Pharmaceutical Industry Perspective:a New Era in Separation Science
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Background: Pharmaceutical R&D productivity
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Pammolli et al, Nature Rev. Drug Discov., 2011, 10, 428
10 years in 1990s
14 years after 2000
Longer development time
Avg. time for products launched
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Complex emerging drug discovery paradigm
3Valeur et al, Angew. Chem. Int. Ed. 2017, 56, 1029
Examples…
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Complex drug delivery technologies
Kolate et. al. J Control. Release, 2014, 192, 67
4
Chang et al, J Pharm Sci., 2015, 104, 3404
Long Acting drug Release Fab-PLGA
Procko et al, Cell, 2014, 157, 1644
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It takes too many tools and too long to characterize one sample – Need multi-attributes characterization simultaneously
5
And more…
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Challenges in pharmaceutical HPLC analysis
K. Zhang et. al., Am. Pharm. Rev., 2013, 16 (7), 39-44.
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Better Separation Science
• Material
• Instrumentation
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Material: e.g. mixed-mode, “total separation”
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Ideally, a column or such a material that…
Although not yet to make this total separation, mixed-mode column technology has been greatly advanced in the past few years.
Mixed-Mode ChromatographySample Mixed-mode Stationary Phase
K Zhang, X, Liu, JPBA, 2016, 128, 73
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Mixed-Mode Stationary Phases
8X. Liu, C. Pohl, J. Sep.Sci. 33 (2010) 779–786.
RP/IEX bimodal mixed-mode phases
RP/AEX/CEX trimodal mixed-mode phases Restricted access
GE Capto Core 700 particle
BimodalRP/AEXRP/CEXHILIC/AEXHILIC/CEXTrimodalRP/AEX/CEX HILIC/AEX/CEX
K Zhang, X, Liu, JPBA, 2016, 128, 73
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Generic method for 25 common counterions using mix-mode column coupled with CAD
Ca
Lact
ate
Proc
aine
TRIS
K
NaChol
ine
Meg
lum
ine
NO3
Mal
eate
Gluc
onat
e
BrCl
Tosy
late
Succ
inat
e
Besy
late
Zn
Mal
ate
PO4
Fum
arat
eTa
rtra
teCi
trat
e
Mes
ylat
eSO4
Mg
9K Zhang, L Dai, N Chetwyn, J. Chromatogr. A. 1217 (2010) 5776-5784
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Applications in drug, counterion and trace level residual metal analysis
10L. Dai, L. Wigman, K. Zhang, J Chromatogr. A, 2015, 1408, 87
API
Li
LOQ: 8 ppb (w/w) in sample matrix
API
Fumarate
NaproxeneNa
Adenine
Cl
Separation of API and counterions
K Zhang, L Dai, N Chetwyn, J. Chromatogr. A. 2010, 1217, 5776
Sensitive quantification of Lithium in drug substance by HILIC separation
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Instrumentation:On-line multidimensional UHPLC with hyphenated detection
K Zhang, et al, J. Sep. Sci. 2013, 36, 2986
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Analysis of unconjugated small molecules in ADC by 2D-LC/MS
Y. Li et al, J. Chromatogra. A, 2015, 1393, 81–88.12
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2D-LC/CAD-MS characterization and stability study of polysorbate in mAb formulation
13Li, Y.; Hewitt, D.; Lentz, Y. K.; Ji , J. A.; Zhang, T. Y.; Zhang, K.;, Anal Chem, 2014, 86, 5150.
2D-LC• Mixed mode (AEX/RP)–RP • CEX-RP
POE isosorbide
POE
POE sorbitan
mAb: 10-100 mg/mL polysorbate: 0.02-0.05%
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2D-LC/MS study of the degradation product and mechanism in microdose PEG formulation
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Microdosing for Bioavailability Study
3µg API in 100mL IV bag: 10% Degradation ~ 3 ppb (w/v) level4% PEG in saline
PVC infusion tubing
L. Dai et al, JPBA, 2017, 127, 182
Co-eluting impurity
IV device
1st Dimension
01
23
45
67
89
Time (min)
10 20 30 40 50 60 70 80 90
100
Relative Abundance
N
O
H3N
R1
Dehydrochlorination on PVC tubing surface and PEG auto-oxidation formed an oxidative environment.
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Trace analysis: genotoxic impurity hydrazine by derivatization
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H2N NH2
O
H
OH+
OH
NN
HO
HNA
406
230
Wavelength (nm)200 300 400 500 600
Inte
nsity
(mA
U)
Ultraviolet Visible
DerivativeStrong Absorbanceλmax = 406 nm
GNE API
AU
0.000
0.002
0.004
Minutes0.00 2.00 4.00 6.00 8.00
Background of drug matrixUV = 260 nm
AU
0.000
0.002
0.004
Minutes0.00 2.00 4.00 6.00 8.00
UV = 406 nm
High sensitivity High resolution No matrix interference
10 ppm Hydrazine
HN
A
API
260
Wang, et al, JPBA, 2016, 126, 141
Matrix interference
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Separation of Compounds with Multiple Chiral Centers by 2DLC
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Drug with 3 chiral centers
2n Rule
e.g, n=3, 8 chiral isomers
It is highly challenging to separate compounds with multiple chiral centers 2DLC strategy
1st D: Well-developed achiral reversed-phase method (typically QC assay/impurity method) separate diastereomers and other impurities 2nd: Chiral method to separate each enantiomers
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Reaction screening, Scavenger
High Throughput Experiment and Automation
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Gas phase reaction
Informatics
Form and Salt selection Crystallization
Excipient Compatibility/Forced Degradation
Solubility
• HPLC analysis is the bottleneck
Absorbent Screening
Chiral,Product isolation
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Summary
• Fast, sensitive, selective, robust separation methods
• New separation materials and instrumentation
• Complex modality characterization that can predict drug activity
• Characterization multiple attributes simultaneously
• Platform technology, less compound specific method development
• Miniaturization
• Green chemistry
• Intelligent software
• Well-trained scientists in separation science
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Acknowledgements
Genentech• Peter Yehl
• Larry Wigman
• Joe Pease
• Colin Masui
• Geoffrey Yeh
• Francis Gosselin
• Nik Chetwyn
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