Lyophilization Validation:

A Regulatory Perspective

Ellen Huang
CBER/OCBQ/DMPQ
CASSS CMC Strategy Forum
July 19, 2016
1
 Overview
• Objective
• Definition of lyophilization
• Observations and challenges
• Lyophilization process validation
• Aseptic processing
• Cleaning and sterilization
• Orphan products
• Alternative containers
2
 Objective
• The objective of this presentation is to
present an overview of FDA’s expectations
for validating the lyophilization process

3
 What is Lyophilization
• A process in which water is removed from
a product after it is frozen and placed
under a vacuum, allowing the ice to
change directly from solid to vapor without
passing through a liquid phase.
• The process consists of three separate,
unique, and interdependent processes;
freezing, primary drying (sublimation), and
secondary drying (desorption). 4
 Annealing
• Optional step(s), that typically follows the
freeze step where the product is warmed up
to allow crystals to grow
• Performed because crystalline component
may not be completely crystallized
– Provides necessary cake structure or more
stability to the protein
• Facilitates faster sublimation, thus
optimizing the process
5
 Lyophilization
• In general, lyophilization is used for drug
products to provide for greater stability and
increase the product’s shelf life.
• Prior to use, the product is reconstituted
with the appropriate diluent.
• Lyophilization is sometimes used on drug
substance, especially for long-term
storage, e.g., multiple conjugates
6
 Observations and Challenges
• Establishing a commercial lyophilization
process can be challenging
• Successful validation requires robust
development studies, equipment
qualification, and process validation
• Not enough focus on developing the
freezing phase, the most important phase
of the cycle
7
 Observations and Challenges
• Inappropriate bracketing strategies (e.g., load
size or number of lyophilizers)
• Inadequate empty chamber shelf temperature
and product temperature mapping
• Insufficient sampling for quality and uniformity
– Sample locations unknown
• Worst-case load not repeated
• Scale-up or technical transfer issues
• Vial/container imperfections 8
 Equipment Qualification
• Empty chamber temperature mapping
– Thermocouples (TC) are typically placed in
the four corners and center of each shelf
– Identify cold and hot spots
– Expect limited variability on each shelf and
between shelves (usually about 1-2°C)
– Temperature range should exceed actual
lyophilization temperatures
• Leak rate test
9
• Condenser capacity
The Process Validation Continuum
Pre-IND IND Phase I Phase II Phase IIb Phase III BLA Post-marketing

Stage 3: Continued
Stage 2: Process
Stage 1: Process Design through Process Verification
Qualification that is
iterative risk assessment, quality  Continuous
sufficiently
characterization and process process and
comprehensive and
characterization cycles, with the product monitoring
robust to provide
objective of establishing a commercial conclusive evidence  Trending and
process with sufficient control to that the commercial annual product
consistently produce drug substance process as designed reviews
and drug product meeting defined consistently achieves  Deviation
specification  manufacturing phases specified product management
linked through comparability quality in the  Change control
commercial production  Validated process
environment improvements
 State of control
10
 Stage 1: Process Design
Process Design: The commercial manufacturing
process is defined during this stage based on
knowledge gained through development and
scale-up activities
• Understand the product and critical properties of
the formulation
– Thermal characterization (collapse temperature,
eutectic temperature, and/or glass transition
temperature)
– Stability of the product
– Properties of the excipients used 11
 Development Studies
• Laboratory, pilot, and at-scale scale
studies to support commercial cycle
• Design of experiment studies to
understand impact of parameters (e.g.,
shelf temperature, pressure, time, and
ramp rate) and design space for the
product

12
 Stage 2: Process Qualification
At this stage, the process design is evaluated to
determine if the process is capable of reproducible
commercial manufacturing.
– Combines qualified facility, equipment, utilities, and
trained personnel with the Process Performance
Qualification (PPQ) (may include at-scale engineering
runs)
• To demonstrate product uniformity and the ability
to repeat and consistently manufacture product.
• Product uniformity is demonstrated through product
temperature mapping and extended
13
sampling/testing of reconstituted product.
 Product Temperature Mapping
• TC are typically placed in the product in the four
corners and center of each shelf
– Can use “seeded” runs
– Use of an appropriate surrogate
• Temperature profile diagrams useful in
determining when primary and secondary drying
have completed
– Need to add additional time to primary drying since
containers with TC nucleate at higher temperature
• Usually performed as engineering runs since
using TC throughout the load is not aseptic
14
 Temperature Profile Diagram

Freezing Primary Drying Secondary

Drying
Trappler, E. H. (2007). Validation of Lyophilization. In F. J. James P. Agalloco, 15
Validation of Pharmaceutical Processes (p. 388). CRC Press.
 Extended Sampling
• Sample vials typically from four corners
and center of each shelf
• Sample locations should be known
• Testing often includes residual moisture,
reconstitution time, cake appearance (no
collapsed cake or melt back), and
reconstitution appearance,
• Can be performed as part of PPQ /
conformance runs 16
 Cake Appearance
Discernible Effects - Discernible Effects -
Syringes Vials

PDA Course No. 282, Validation of Lyophilization (May 2015)

17
 Boundary Studies
• Studies where shelf temperature and
chamber pressure were offset from
commercial set-points
– High and low shelf temperature
– High and low chamber pressure
• Sometimes time is also varied during
these studies
• Not required for fixed cycles but provides
confidence in lyophilization process 18
 How many runs?
• It depends
– Typically three runs are performed on the
maximum load and one run is performed on
the minimum load
– How much supportive development work
– How many factors there are (e.g., number of
lyophilizers, dosage strength, etc.) and how
study is bracketed

19
 Bracketing Strategy
• Load size (maximum and minimum)
• Multiple lyophilizers
– Demonstrate equivalence between the units
– Typically 3+1+1
• Multiple dosage strengths
– Consider thermal characteristics between
dosage strength
• Fill volume
• Different vials size or manufacturers 20
 Stage 3: Continuous Process
Verification (CPV)
CPV: Ongoing assurance is gained during routine
production that the process remains in a state of
control.
• Continued monitoring and sampling of process
parameter and quality attributes
– Analyze and trend the data
– Once sufficient data has been gathered, can reduce
monitoring with justification
– Monitor changes in process inputs, including materials
and container/closure components
• Imperfections in the vial can have negative impact on crystal 21
formation during freezing.
 Aseptic Processing
• Transportation of vials from the filling line to
the lyophilizer should be done aseptically
since vials are partially stoppered
• Media challenges should include
transportation, loading, holding, partial
vacuum, stoppering, and unloading the
lyophilizer, however:
– Media should not be frozen as this may kill
organisms
– Hold time does not need to be the actual duration
22
of lyophilization cycle
 Cleaning
• Perform between each run
• Clean-In-Place (CIP) or manual cleaning
– CIP cycle: initial rinse, recirculation, final rinse,
drying
– CIP CV should demonstrate total chamber
coverage (riboflavin)
• WFI is preferred
– If cleaning agent is used, must demonstrate
removal from the chamber
• Cleaning process should be validated
– Use worst-case soil
23
– Procedure for potential spills
 Sterilization
• Lyophilizer should be sterilized after
cleaning
• Typically observe steam sterilization (SIP)
(overkill approach)
• Sterilization process should be validated
– Heat distribution and biological indicators
– Demonstrate Sterility Assurance Level of 10-6
• Chamber should be dry after SIP
24
 Orphan Products
• Use of an appropriate surrogate or
perform with seeded runs
• Less PPQ runs may be submitted in
submission since product is manufactured
infrequently
– Perform additional runs in the future to
support the PPQ
• No exemption from CGMP compliance
25
 Alternative Containers: Trays
• Membrane Trays (e.g., LYOGUARD®)
– Tray with a PTFE membrane
– Usually has low fill depth -> Larger shelf
contact area
– Temperature mapping requires more than one
TC per container
– Typically seen used for drug substance

26
Alternative Containers: Syringes
• Pre-Filled Syringes and Dual Chamber
Syringes
• Syringe has less/no contact with the shelf, so
heat transfer will be different than a vial.
Therefore, syringes respond more slowly to
shelf temperature changes.
• Consider type of syringe holder and loading
configuration
• Validation expectation same as vial
27
 Acknowledgements
• Nicole Trudel
• Randa Melhem, PhD
• Qiao Bobo, PhD
• Tony Lorenzo
• Debbie Trout
• Jay Eltermann
• Laurie Norwood
• Mary Malarkey 28
 Definitions
• Collapse Temperature (Tc) – The temperature
above which the frozen formulation undergoing
lyophilization loses macroscopic structure and
collapses; below this temperature, the
solute/solvent mixture is able to retain its
structure, even when it is not supported by
surrounding ice, which is removed via
sublimation during primary drying.
• Eutectic Temperature (Te) – The point at which
the product only exists in the solid phase,
representing the minimum melting temperature.
29
Not applicable to amorphous material.
 Definitions
• Glass Transition Temperature (Tg or Tg’) – the
temperature above which the formulation
undergoing lyophilization
• Metastable – pertaining to a body or system
existing at an energy level above that of a more
stable state and requiring the addition of a small
amount of energy to induce a transition to the
more stable state.
• Sublimation – The transition of a substance
(such as ice) from a solid state directly to the
vapor state (such as water vapor) without first 30
passing through an intermediate liquid phase.