introducing milligram screening reactions into the kilogram world of chemical development simon...
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Introducing Milligram Screening Reactions into the Kilogram World of Chemical Development
Simon Yates, AstraZeneca.
FreeSlate European User Meeting24th September 2013
Our Journey so far…
• We produce a scalable process, not just a compound
• From a few grams in the lab to multi-kilo plant campaigns
• Emphasis on SELECT criteria• Safety• Economics • Legal• Environment• Control• Throughput
What is Chemical Development?
In number of metal catalysed reactions in Med Chem
The time to develop process for early compounds
Development time by using scavengers
100’SOf scavengers and carbons to choose from
Time, effort and missed opportunity by automated screening
Reproducibility by screening under inert conditions
Scavenging Challenge?
Transition Metals are toxic!
Conventional work up takes time to develop
But what kit?
• 10 reactions at a time• Minimum volume ~2mL
Integrity 10
MT Flexiweigh 2x MT Mini Mapper
But what kit?
• Inerted Glovebox • 20mL reaction tubes• 100mg reactions• 24 wells
• Very manual• Lots of programming
In number of reactions we could run (up from 24)
Manual intervention, to make this as routine as possible
Ease of programming and data analysis
‘Future proof’ our investment
Reaction scale
Wanted to….
Purchased a Symyx CM2
Solid Dispensing
9 heat-stir bays
Vial gripper16G to 20G needle (with N2 pressure)- 1mL & 500uL syringes attached to 10 off deck solvents
10G needle(5mL syringe)
Bespoke filter equipment- Symyx filter too small
Scavenging Workflow
96 x 1mL vials ~40 different scavengers
Seal, heat, stir 16 hours
Stock of reaction solution
Centrifuge for5 min
Scavenging Workflow
Air GapBacking Solvent
Overshoot
ICP sample
HPLC sample
RAS
ICP = Inductively Coupled Plasma
• We have run 20+ screens and saved projects time and money
• Numerous examples of where scavengers used in all scales of manufacture
• Pd is most common metal scavenged, now have a generic plate of 22 scavengers.
• Improvements in whole workflow reduced time from 5 working days to 2 days.
Scavenging today
Cross Coupling
B
Br
+
OH
OH
Pd Ligand Base(Internal Std)
R1-M R2-X R1-R2
Background
Original plan was to run X-Coupling
Timing forced by closure of our existing facility
In number of reactions we could runcover more experimental space, quicker.
Reaction scale
Apply our previous learning
Pre-screening~10 reactions
Optimisation of continuous parameters
DoE (24-plate)
96-well plate screening
Discovery
Life Cycle Managemen
tDevelopment
96-well plate screening
Optimisation of continuous parameters
DoE (24-plate)
What we developed
18
• Developed ‘Generic plates’ for 5 different reaction types • Based on literature and in house expertise
Generic Reaction Plates
19
Manual Pipette
Evaporate off carrier solvent
Stock solutions of- Ligands- Metals - Internal std
Generic plate formation
-or - Automated dispensing and store
Running a project…
Define:Bases and Solvents
Add in solid(s)Reactants
Add in reaction solvent
Quickly add in Aq. Base‘Start of reaction’
Seal upHeat and Stir
2 x Manual Sample Prep ~2hrs and 20hrs
Running a project…
1st sample will always be manual 2nd sample could be automated – middle of night.
Future improvements
Reactions run on 200-400uL scale at 10C below bp
Solvent loss / corrosion of sealing material
Fully closed plate
Can’t sample by CM2
Need a pierceable, but re-sealable, membrane
Any ideas welcome!!
Material consumption
24 well - DoE type
Expanding on 96 well hit(s)
Continuous variables
BA
Discrete variables
Profile reactions Statistical Analysis
Catalyst Plate
Reaction Plate
24 well – DoE type
24 x 4mL vials Weigh inLigandsMetal
Add reaction solvent
Cap, heat and stir, 60min.
24 x 4mL vials Weigh inReactants, BasesInternal Standard
Add reaction solvent/liquid reagents
Reaction Plate
24 well – DoE type
Transfer from Catalyst plate to reaction plate
Heat and Stir Automatically sample 4 times over 16 hours
Run LC-Mass Spec on samples
Future improvements
1 Temperature (plate) per runLose a key factor
Multiple Plates / run sampled into 1 HPLC plate
Future improvements
Sampling always run at the end, not good for fast reactions
Allow sampling as part of ‘dispense’
And/or move vials to heat zonesto ‘start’ reactions(Josh Denette and Kristin Price at Pfizer)
Data Handling
RAS
MDB
29
Asymmetric Hydrogenation
Successfully built capability in Sodertalje, Sweden
site end of 2012
Opportunity to purchase new equipment
CM3 (in glovebox) and SPR Off deck hydrogenation – make use of CM3 during long reactions
Knowledge of LEA / CM2
CM2 / CM3 used as 1 resource for X-Coupling and Asymm. Hydrog.
Commissioning - NOW
Conclusion
We have come along way since 2007
Scavenger was the warm up act
• Scavenging and X-Coupling saved millions of dollars already • Success led the way to keep Asymm. Hydrog. in house • Asymm. Hydrog. will have similar if not bigger impact
Continue to develop workflows
Keep control of the data96 x 2 x 5 x 50 = 48K data points / year
Conclusion
User community carry on sharing and learning What’s good for me, could be good for you too.
Acknowledgments
AstraZenecaJohn Leonard, Gair Ford, Barney SquiresPhil Hogan, Keith Mulholland, Andy CampbellRachel Munday, Kevin LeslieSarah Thompson, Andy Poulton
Ex AstraZenecaPaul Murray and co. (catsci.com)Per Ryberg and co. (SP Technical Research Institute of Sweden)
Symyx/FreeSlateSteve Yemm, Zack Hogan, Colin Masui, Anny Tangkilisan, Rob Rosen, Eric CarlsonPeter Huefner, Grant Gavranovic, Justin Fisher, Jonathan HarrisPeter Gravil, Jos De Keijzer, Rick Sidler, Tony Mani, Guillaume Magan, Ludovic Edvard
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