-
The NanoAssemblr™ Platform: Microfluidics-Based Manufacture
of
Nanomedicines
Precision NanoSystems, Inc. September 2014
-
Conceptual Nanoparticle
Scale-up Bench-Scale NanoparticleFormulation
Manufacturing Process
Robust Manufacturing
Process
Nanomedicine Product
Nanoparticle Development Process
2
-
Conceptual Nanoparticle
Scale-up Bench-Scale NanoparticleFormulation
Manufacturing Process
Robust Manufacturing
Process
Nanomedicine Product
Nanoparticle Development Process
3
NanoAssemblr™ Benchtop Instrument
-
Conceptual Nanoparticle
Scale-up Bench-Scale NanoparticleFormulation
Manufacturing Process
Robust Manufacturing
Process
Nanomedicine Product
Nanoparticle Development Process
4
NanoAssemblr™ Benchtop Instrument
Accelerated Development of Nanoparticles
Scale-Up Platform
-
5
ü Proprietary microfluidics-based instrument
ü Rapid nanoparticle prototyping
ü Single step
ü Small sample volumes
ü Software controlled, automated
ü Intra- and inter-operator reproducibility
ü Robust processing and variable manipulation
The NanoAssemblr™ Benchtop Instrument
Microfluidic Cartridge
NanoAssemblr™ Benchtop Instrument
-
6
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
The Magic is in the Microfluidics
Exquisite Control Over Manufacturing Process
organic aqueous
Rapid & Controlled Mixing
Mixer design by: Stroock et al., Science 2002
Channel diameter: ~100 µm
Staggered Herringbone
Mixers
Nanoparticles
-
The NanoAssemblr™: Manufacture of Novel Nanoparticles
O/W Nanoemulsions
7
Lipid Nanoparticles
Liposomes
Polymer Nanoparticles
-
8
0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 10 20 30 40 50 60 70 80 90
Actual diameter Theoretical diameter
Part
icle
siz
e, n
m
POPC/Triolein ratio, mol/mol
Zhigaltsev, I.V. Et al., Langmuir 2012, 28, 3633−3640
Nanoemulsion Droplet Size Dictated by Emulsion Composition
-
Nanoemulsion Droplet Size Dictated by Manufacturing Process
9
0 1 2 3 4 5 6 7 8 9 10
10
20
30
40
50
60
70
B POPC/triolein (60/40 mol/mol)
Par
ticle
siz
e, n
m
Aqueous/ethanol flow rate ratio
Zhigaltsev, I.V. Et al., Langmuir 2012, 28, 3633−3640
-
10
Liposome Size Dictated by Manufacturing process
“Limit size” Liposomes are dictated by the Flow Rate Ratio
1 2 3 40
20
40
60
80
100
Flow Rate Ratio (Aqueous:Organic)
Z-A
ve (n
m)
POPC:Chol:DSPE-PEG (55:42:3)
DSPC:Chol:DSPE-PEG (55:42:3)
Post-Dialysis DataAll PDI < 0.09
-
Liposome Size Dictated by Lipid Composition
11
0 10 25 35 4220
30
40
50
0.00
0.05
0.10
0.15
0.20
0.25
Cholesterol content [mol%]
Z-A
ve
[nm
]
PD
I
Liposome size and size distribution is dependent on Cholesterol
content
POPC:Cholesterol:PEG-DSPE (3%)
-
RNA-Lipid Nanoparticle Size Dictated by Lipid Composition
12
Dia
me
ter
(nm
)
PEG-Lipid Content (mol %)
10
20
30
1.0 2.5 5.00
40
50
60
PD
I
0.02
0.04
0.08
0.00
0.10
0.06
“Limit Size” is Dependent on RNA-Lipid Nanoparticle
Composition
-
13
RNA-Lipid Nanoparticle Size Dictated by Manufacturing
Process
siRNA-LNP (Cationic Lipid:DSPC:Cholesterol:PEG) Changing
Process
Dia
me
ter
(nm
)
Flow Rate (mL / min)
20
40
0 120
60
80
100
4 8 16 20 24
RNA-Lipid Nanoparticles Reach “Limit Size” at High Flow
Rates
-
CellaxTM Polymer-Drug conjugates
14
100nm 100nm
NanoAssemblr TM Vortex
-
4 8 15 20 30 35
60
80
100
120
0.0
0.1
0.2
0.3
concentration [mg/g]
Z-A
ve
[nm
]P
DI
Nanoparticle Size Dictated by Polymer Concentration
15
Particle size is dictated by polymer concentration
CellaxTM Polymer-Drug conjugates
-
16
10 12 14 16 18 20 22 24 26 28 30 320.0
0.1
0.2
0.3
Flow-Rate [ml/min]
PD
I
Polymer Nanoparticle Size Distribution Dictated by Manufacturing
Process
Polydispersity is dependent on flow rate
CellaxTM Polymer-Drug conjugates
-
Reproducible RNA-Lipid Nanoparticles Independent of Operator or
Site
17
Automated Instrumentation Removes Operator Variability
Dia
me
ter
(nm
)
Sample
20
40
1 40
60
2 3 5 6
PD
I
0.04
0.12
0.00
0.16
0.08
Operator
-
Assessment of the Robustness of the Manufacturing Process
18
Stable Results = Robust Process = Scalable Process
Design of Experiment (DoE) variables – Lipid Concentration –
Flow Rate – Mixing Conditions – Lipid:RNA Ratio
-
19
Continuous Flow Pumps
Parallelized Microfluidic Mixers
RNA -Nanoparticles
Aqueous (RNA)
Solvent (Lipids)
Microfluidic Mixer
Microfluidic Mixer
Microfluidic Mixer
Microfluidic Mixer
Microfluidic Mixer
Dilution Buffer Exchange
& Nanoparticle
Concentration
Microfluidics Enables “Seamless Scale-Up”
-
20
Continuous Flow Pumps
Parallelized Microfluidic Mixers
RNA -Nanoparticles
Aqueous (RNA)
Solvent (Lipids)
Microfluidic Mixer
Microfluidic Mixer
Microfluidic Mixer
Microfluidic Mixer
Microfluidic Mixer
Dilution Buffer Exchange
& Nanoparticle
Concentration
Microfluidics Enables “Seamless Scale-Up”
-
Continuous Flow Scale-up Manufacture of RNA-Lipid Nanoparticles
Using Single Mixer
21
Seamless Transfer of Optimized Manufacturing Parameters
0.00
0.02
0.04
0.06
0.08
0.10
0
10
20
30
40
50
60
PD
I
Dia
me
ter
(nm
)
Cumulative Fraction (mL)
-
22
Continuous Flow Pumps
Parallelized Microfluidic Mixers
RNA -Nanoparticles
Aqueous (RNA)
Solvent (Lipids)
Microfluidic Mixer
Microfluidic Mixer
Microfluidic Mixer
Microfluidic Mixer
Microfluidic Mixer
Dilution Buffer Exchange
& Nanoparticle
Concentration
Microfluidics Enables “Seamless Scale-Up”
-
-0.02
0
0.02
0.04
0.06
0.08
0.1
0
10
20
30
40
50
60
1X Mixer 2X Mixer 4X Mixer
PD
I
Dia
me
ter
(nm
)
Manifold Microfluidic Parallelization Enables RNA-Lipid
Nanoparticle Scale-Up Manufacture
23
12 mL/min 24 mL/min 48 mL/min
Parallelization of Microfluidic Mixers Enables Higher
Throughput
-
On-Chip Microfluidic Parallelization Produces Equivalent
RNA-LNP
24
0
0.02
0.04
0.06
0.08
0.1
0
10
20
30
40
50
60
4X Manifold 4X On-Chip
PD
I
Dia
me
ter
(nm
)
Multiple options for increased throughput by parallelization
48 mL/min 48 mL/min 4X Mixer Chip
-
25
Aqueous Metering Pump
Solvent Reagent Bag
Aqueous Reagent Bag
Solvent Metering
Pump
Dilution Pump
Dilution Reagent Bag
Pinch Valve
To Post-processing
Tee
Tee
Pinch Valve
Sample Switch Waste
288 mL/min
384 mL/min
8X Scale-up System
8 x 12 mL/min
24 mL/min
72 mL/min 96 mL/min
Microfluidic Mixer Array
Design for GMP Manufacturing
-
8X Scale-Up Instrumentation Produces High-Quality RNA-LNP
26
0.00
0.02
0.04
0.06
0.08
0.10
0
10
20
30
40
50
60
PD
I
Dia
me
ter
(nm
)
Cumulative Fraction (mL)
• 8X Scale-Up System Processes 5.75 L/hr • Further
parallelized systems under development
-
GMP Program in Development
27
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
-
Try the NanoAssemblr™ for your drug development program
ü Worldwide distribution
ü Demo Program Available ü Contact Colin Walsh:
[email protected] ü Contact Gesine Heuck:
[email protected]
ü Development programs in place
ü RNA delivery systems ü Liposomes ü Polymers ü Polymer-drug
conjugates ü Other nanoparticle systems
28
NanoAssemblr™ Benchtop Instrument
