The NanoAssemblr Platform: Microfluidics-Based Manufacture of Nanomedicines. Precision NanoSystems, Inc. September 2014
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1 The NanoAssemblr Platform: s-based Manufacture of Nanomedicines Precision NanoSystems, Inc. September 2014
2 Nanoparticle Development Process Conceptual Nanoparticle Bench-Scale Nanoparticle Formulation Manufacturing Process Robust Manufacturing Process Scale-up Nanomedicine Product 2
3 Nanoparticle Development Process Conceptual Nanoparticle Bench-Scale Nanoparticle Formulation Manufacturing Process Robust Manufacturing Process Scale-up Nanomedicine Product NanoAssemblr Benchtop Instrument 3
4 Nanoparticle Development Process Conceptual Nanoparticle Bench-Scale Nanoparticle Formulation Manufacturing Process Robust Manufacturing Process NanoAssemblr Benchtop Instrument Scale-up Nanomedicine Product Scale-Up Platform Accelerated Development of Nanoparticles 4
5 The NanoAssemblr Benchtop Instrument ü Proprietary microfluidics-based instrument ü Rapid nanoparticle prototyping ü Single step NanoAssemblr Benchtop Instrument ü Small sample volumes ü Software controlled, automated ü Intra- and inter-operator reproducibility ü Robust processing and variable manipulation Cartridge 5
6 The Magic is in the s aqueous organic Channel diameter: ~100 µm Staggered Herringbone s Nanoparticles Rapid & Controlled Mixing design by: Stroock et al., Science 2002 ü Laminar flow controlled mixing ü Rapid mixing (3 ms -1 ) ü Nanoliter Reaction volumes: ~ 14 nl ü Low energy input, low shear system ü Seamless scale-up Exquisite Control Over Manufacturing Process 6
7 The NanoAssemblr : Manufacture of Novel Nanoparticles O/W Nanoemulsions Liposomes Lipid Nanoparticles Polymer Nanoparticles 7
8 Nanoemulsion Droplet Size Dictated by Emulsion Composition Actual diameter Theoretical diameter Particle size, nm POPC/Triolein ratio, mol/mol Zhigaltsev, I.V. Et al., Langmuir 2012, 28,
9 Nanoemulsion Droplet Size Dictated by Manufacturing Process B 70 POPC/triolein (60/40 mol/mol) Particle size, nm Aqueous/ethanol flow rate ratio Zhigaltsev, I.V. Et al., Langmuir 2012, 28,
10 Liposome Size Dictated by Manufacturing process DSPC:Chol:DSPE-PEG (55:42:3) POPC:Chol:DSPE-PEG (55:42:3) Z-Ave (nm) Post-Dialysis Data All PDI < Flow Rate Ratio (Aqueous:Organic) Limit size Liposomes are dictated by the Flow Rate Ratio 10
11 Liposome Size Dictated by Lipid Composition Z-Ave [nm] POPC:Cholesterol:PEG-DSPE (3%) PDI Cholesterol content [mol%] Liposome size and size distribution is dependent on Cholesterol content 11
12 RNA-Lipid Nanoparticle Size Dictated by Lipid Composition Diameter (nm) PDI PEG-Lipid Content (mol %) 0.00 Limit Size is Dependent on RNA-Lipid Nanoparticle Composition 12
13 RNA-Lipid Nanoparticle Size Dictated by Manufacturing Process Changing Process sirna-lnp (Cationic Lipid:DSPC:Cholesterol:PEG) 100 Diameter (nm) Flow Rate (ml / min) RNA-Lipid Nanoparticles Reach Limit Size at High Flow Rates 13
14 Cellax TM Polymer-Drug conjugates 100nm 100nm NanoAssemblr TM Vortex 14
15 Nanoparticle Size Dictated by Polymer Concentration Cellax TM Polymer-Drug conjugates Z-Ave [nm] PDI concentration [mg/g] 0.0 Particle size is dictated by polymer concentration 15
16 Polymer Nanoparticle Size Distribution Dictated by Manufacturing Process 0.3 Cellax TM Polymer-Drug conjugates 0.2 PDI Flow-Rate [ml/min] Polydispersity is dependent on flow rate 16
17 Reproducible RNA-Lipid Nanoparticles Independent of Operator or Site Diameter (nm) PDI Sample Operator Automated Instrumentation Removes Operator Variability 17
18 Assessment of the Robustness of the Manufacturing Process Design of Experiment (DoE) variables Lipid Concentration Flow Rate Mixing Conditions Lipid:RNA Ratio Stable Results = Robust Process = Scalable Process 18
19 s Enables Seamless Scale-Up Aqueous (RNA) Solvent (Lipids) Continuous Flow Pumps Dilution Buffer Exchange & Nanoparticle Concentration RNA - Nanoparticles Parallelized s 19
20 s Enables Seamless Scale-Up Aqueous (RNA) Solvent (Lipids) Continuous Flow Pumps Dilution Buffer Exchange & Nanoparticle Concentration RNA - Nanoparticles Parallelized s 20
21 Continuous Flow Scale-up Manufacture of RNA-Lipid Nanoparticles Using Single Diameter (nm) PDI Cumulative Fraction (ml) Seamless Transfer of Optimized Manufacturing Parameters 21
22 s Enables Seamless Scale-Up Aqueous (RNA) Solvent (Lipids) Continuous Flow Pumps Dilution Buffer Exchange & Nanoparticle Concentration RNA - Nanoparticles Parallelized s 22
23 Manifold Parallelization Enables RNA-Lipid Nanoparticle Scale-Up Manufacture Diameter (nm) PDI X 2X 4X 12 ml/min 24 ml/min 48 ml/min Parallelization of s Enables Higher Throughput 23
24 On-Chip Parallelization Produces Equivalent RNA-LNP Diameter (nm) PDI 4X Chip 0 4X Manifold 4X On-Chip 48 ml/min 48 ml/min 0 Multiple options for increased throughput by parallelization 24
25 Design for GMP Manufacturing 8 x 12 ml/min Array 288 ml/min 24 ml/min Dilution Pump Solvent Metering Pump Dilution Reagent Bag Tee Pinch Valve 384 ml/min To Post-processing Solvent Reagent Bag 72 ml/min 96 ml/min Tee Aqueous Metering Pump Pinch Valve Aqueous Reagent Bag Sample Switch Waste 8X Scale-up System 25
26 8X Scale-Up Instrumentation Produces High-Quality RNA-LNP Diameter (nm) PDI Cumulative Fraction (ml) 8X Scale-Up System Processes 5.75 L/hr Further parallelized systems under development 26
27 GMP Program in Development ü Prototype instrumentation developed ü Disposable COC microfluidic chips developed ü Working with drug development partner to transfer technology to CMO for scale-up and GMP manufacturing ü Fully disposable fluid path using USP Class 5/6 materials ü Working with partners and to support development and push programs forward 27
28 Try the NanoAssemblr for your drug development program ü Worldwide distribution ü Demo Program Available ü Contact Colin Walsh: ü Contact Gesine Heuck: ü Development programs in place ü RNA delivery systems ü Liposomes ü Polymers ü Polymer-drug conjugates ü Other nanoparticle systems NanoAssemblr Benchtop Instrument 28
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