Principles and Applications of Lyophilization to Biotechnology

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6 Δεκ 2012 (πριν από 4 χρόνια και 9 μήνες)

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1

Baxter Confidential


Principles and Applications of
Lyophilization to Biotechnology

Michael J. Akers, Ph.D.

Baxter
BioPharma

Solutions

Bloomington, Indiana

BIOMAN 2010 Conference

Ivy Tech Community College

Bloomington, Indiana

July 13, 2010

Organization of Presentation

Baxter Briefs

Biologics
Market

Lyophilization

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Baxter Confidential

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Baxter Confidential

Bloomington, Indiana
-

Capabilities Overview


Prefilled syringes


Lyophilization


Aseptic liquid vials


Cartridges


Diluent

syringes

Technologies Services


Formulation development


Process development


Lyo

cycle development


Analytical development


Regulatory support


Labeling and Packaging


Clinical to Commercial

Other Interesting Facts about

Baxter BioPharma in Bloomington


Produce more prefilled syringes than any
other manufacturer in North America


Small molecules, vaccines, proteins,
diluents


Produce
injectables

marketed throughout
the world


No history of FDA Warning Letters


308 employees in 2001; 993 end of 2009


Newest manufacturing building won ISPE
facility of the year in 2006

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Baxter Confidential

Focus of Bloomington R&D


Formulation Development of Small Volume Injectables


Liquid


Lyophilized


Extensive experience with



Proteins


Monoclonal Antibodies


Small Molecules


Analytical Method Development


Method development and validation services available


Responsible for validation of transfer of all incoming QC
methods for Bloomington facility


Expertise in cleaning validation limit calculations and method
development/validation


Development stability studies


Lyophilization Process Optimization Research Programs


Approximately 80%

of projects in past 2 y

We characterize APIs by thermal analyses and
freeze
-
dry microscopy to develop formulations
and
lyophilization

cycles

We use freeze
-
dry microscopy and FTIR to
evaluate formulation effects on protein stability

We coordinate transfer and scale up of lab
-
developed product to production freeze
-
dry
operations.

Our research programs include mechanisms of
heat and mass transfer during freeze
-
drying,
nucleation optimization, process monitoring by
Tunable Diode Laser Absorption Spectroscopy

Since 2001, 35 peer
-
review research articles, 3
books, 5 book chapters, dozens of short courses,
and 80 presentations at conferences and
universities

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Baxter Confidential

Bloomington R&D Known As Baxter
Lyophilization Center of Excellence

DSC Thermogram of EC90
-60
-55
-50
-45
-40
-35
-30
-25
-20
-15
-10
-5
0
5
10
15
20
25
Temperature, °C
Heat Flow, W/g
Endotherm Up
-30
-28
-26
-24
-22
-20
-18
-16
2 W/g
20 W/g

D
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Baxter Confidential



Baxter BioPharma Solutions


Pharmaceutical Research and Development

Bloomington, IN



Top: Tim Paul, Lisa Hardwick, Nathan Pease, Wendy Saffell
-
Clemmer, Larry Callahan

Middle: Rhonda Haley,Wei Kuu, Melissa Beard, Mike Hildreth, Karen Abram, Kevin O’Bryan, Elizabeth Oslos, Michael Akers

Bottom: Lindsay Stanford, Angie Kruszynski, Nathaniel Fair, Jackie Karty, Greg Sacha, Hoang Mitchell, Dan Staples, Kelly Rob
y,
Steven Nail


Baxter Confidential

8

Market segment by therapeutic area

and molecule class

Oncology
AIID
Diabetes and endocrinology
Infectious disease
Hematology
CNS
Cardiovascular
Women's health
Vaccines
Respiratory
Gastrointestinal
Other
0
5,000
10,000
15,000
20,000
25,000
30,000
35,000
40,000
45,000
Sales ($M)
Market Size of Biological
mAb Sales 2010 ($m)
Protein Sales 2010 ($m)
Total
Source: Baxter Analysis

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Baxter Confidential

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Current/Future Facts About Biologics


Significant growth


2005 sales: $33B


2005 predicted 2010 sales: $82B


1 out of 4 new drugs introduced in US/EU is a
biopharm

product


Monoclonal antibodies (
MoAbs
)


$30B of 2010 sales expected to be
MoAbs


> 200
MoAbs

in clinical development right now


About half of current and new biologic products require
lyophilization


Many of these products will be self
-
administered


Obviously, there is a huge market potential for simple, cost
-
effective devices to be used in home health care to prepare and
deliver lyophilized biologics (and other
lyo

products)


Especially if product is for chronic health care


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Baxter Confidential

Best Known (Best Selling)

Lyophilized

Biologics (Biopharmaceuticals)

Product Brand
(Indication)

Company

2009 U.S. Sales

Enbrel

(Arthritis)

Amgen

$6,580,000,000

Remicade

(Arthritis,
Crohn’s
)

J&J


$5.934,000,000

Herceptin

(Breast cancer)

Genentech


$4.890,000,000

7 Human Growth Hormone
Brands
(Growth hormone
deficiency)

Pfizer, Novo, Lilly,
Genentech,
Serono
,
Sandoz,
Ipsen


$2,886,000,000

Avonex

(Multiple sclerosis)

Biogen

Idec


$2,323,000,000

Advate

(Hemophilia)

Baxter


$2,058,000,000

Betaferon

(
Muliple

sclerosis)

Berlex
/Bayer/Merck


$1,649,000,000

Total Biologic Sales

---

$91,780,000,000

Lyophilization


The terms “lyophilization” and “freeze
-
drying” used
interchangeably


Started in the 1930s, importance grew in WWII


Lyophilization is the conversion of a liquid to a solid
through the process of sublimation


A sterile solution is prepared, filled into primary
containers, placed into a freeze
-
dryer, and frozen


The solvent in the solution is removed by directly
converting it from frozen ice to water vapor.


What remains in the container are the solute
components in the solid state containing very low
residual moisture (typically 0.5
-
2.0%).


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Baxter Confidential

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Baxter Confidential

Dispense raw materials

(active and excipients)

Prepare solution in appropriate

mixing tank (add ingredients to
Water For Injection)

Wash and sterilize

primary containers
and closures

Thaw and pool

active biopharmaceutical

Add active to solution,

pH adjustment, final QS

This is formulation bulk solution

Sterile filter formulated

bulk solution

Aseptically fill formulated bulk solution

into primary package and stopper

(partial stopper if product is to be freeze
-
dried

Transfer to Freeze
-
dryers

and lyophilize

Fully insert stopper, remove
from freeze dryer

Apply aluminum overseal

100% inspection

Storage at appropriate

temperature (usually 2
-
8
ºC)

Label, sec package,
storage, distribution

ISO 8, Grade D,

Class 100,000

ISO 5, Grade A

Class 100

Schematic Overview of Processing
Solution and Freeze
-
Dried
Biopharmaceutical Dosage Forms


Preparing Product for Lyophilization


Prepare the sterile solution

compound,
mix, filter


Fill into containers, typically vials


Partially insert a special designed rubber
closure onto the vials


Aseptically load the vials into a freeze dry
chamber


Freeze every single solution in every vial
below a pre
-
determine critical temperature


Using appropriate application of
temperature and pressure, sublime the ice
from the product


Using further application of temperature and
pressure, remove the necessary amount of
bound water from the product


Automatically stopper the vials, neutralize
the chamber


Aseptically remove the vials from the
chamber and apply aluminum seals


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Baxter Confidential

Today’s State of the Art Technology Uses

Automated Systems To Transfer Product

From Production Line Into the Freeze
-
Dryer

14

Freeze
-
Dryer Subsystems (What Must Be

Operating Smoothly and Maintained for

Lyophilization Process To Succeed?)

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Baxter Confidential

Baxter Pharmaceutical Solutions


Heat Transfer System


Refrigeration System


Vacuum System


Stopper
-
in
-
Place System


Clean
-
in
-
Place System


Control System


Loading/Unloading System

Typical Batch of a Lyophilized Biopharmaceutical Product
Worth Several Million $

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Baxter Confidential

Courtesy of
Samir

U. Sane, Ph.D., Genentech, Inc.

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Baxter Confidential

Product temperature
lower than
shelf temperature

during primary
drying because energy is consumed
as ice sublimes to vapor.

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Baxter Confidential

Flow of Vapor from Chamber to Condenser

T

Connecting Duct

Shelves with
Vials

Vacuum
Pump

Door

Water
Vapor
Flow

T

P

Product
Chamber

Condenser
Chamber

Mushroom Valve

(open position)

Condenser Coils

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Baxter Confidential

Thermal Fluid

Shelf

Frozen Solution

Sublimation Front


Dry Cake

Heat Transfer

Mass Transfer

Schematic of Heat and Mass Transfer

in the Freeze Dryer

Condenser

Vacuum Pump

Thermal fluid circulates within the shelves to
control temperature in chamber

Temperature difference between chamber

and condenser

and pressure differential

between solution in vials and vacuum pump

drives ice out of vial and onto the condenser

Conversion of

solid
(ice) to vapor in
chamber called
sublimation

Pressure gradient between

sublimation front and chamber

ΔP

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Baxter Confidential

Desired Freeze Dried Product Characteristics

Intact cake

Sufficient strength

Uniform color

Sufficiently dry

Sufficiently porous

Sterile

Free of
pyrogens

Free of particulates

Chemically stable

both in dry

state and after reconstitution

Advantages of Lyophilization


Removal of water at low
temperature


Ease of reconstitution


Compatible with aseptic operations


More precise fill weight control


Done properly, the freeze
-
dried
solid has relatively high specific
surface area, which promotes rapid,
complete reconstitution


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Baxter Confidential

Disadvantages of Lyophilization


The drug may not be stable as a
freeze
-
dried solid


Many biological molecules are
damaged by the stresses
associated with freezing, freeze
-
drying, or both


Not all solutes can be freeze
-
dried
to form a pharmaceutically
acceptable cake


Cost may be an issue, depending
on the product


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Baxter Confidential

Common Lyophilized Products


Pharmaceuticals


large and small
molecules


Bacteria


Viruses


Vaccines


Plasma


Small Zoological Specimens
(Taxidermy)


Fruit


Coffee


Flowers


Water
-
Damaged Documents


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Baxter Confidential

Main Challenges of Lyophilization


Development of formulation that meets all the necessary
critical product attribute requirements (quality appearance,
potency, stability, recon time, etc)


Accurate determination of the “critical temperature” of final
formulation necessary to determine conditions of
lyophilization

cycle (
Tg
’, Te,
Tc
)


Establishment of temperature, pressure, and time cycle
settings that can achieve best product quality in shortest
possible time


Transfer and scale
-
up of lab
-
developed process to
production scale process


Keep all the equipment running smoothly


Special challenges in product development of lyophilized
biologic formulations and cycles


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Baxter Confidential

Baxter Pharmaceutical Solutions

Challenges in Lyophilization of Protein
Pharmaceuticals

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Baxter Confidential


Measurable differences in
recovery of activity can be
associated with differences in
the thermal history of freezing


Some proteins can be subject to
overdrying
; that is dryer is not
necessarily better


Stability of the freeze dried solid
is often a concern. Most freeze
dried proteins require
refrigerated storage.


Solid state stability can be
affected by small differences in
residual moisture content.

Challenges in Lyophilization of Protein
Pharmaceuticals

Why Does Lyophilization Take So Long?


Sometimes, the properties of the
formulation require that the
temperature be very low, often
below

30
o
C. This decreases the
driving force


The heat required is very high, and
heat transfer in freeze drying is very
inefficient


Resistance to mass transfer


transport of water vapor from the
sublimation front through the porous
bed of partially dried solids


can be
significant.


Huge batch sizes, takes time to
complete the 3 stages of
lyophilization

Process Data for Representative Freeze-Dry Cycle
-70
-60
-50
-40
-30
-20
-10
0
10
20
30
0
10
20
30
40
50
60
70
Time (hours)
Temperature (°C)
0
50
100
150
200
250
300
Pressure (mTorr)
Thermocouples
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Baxter Confidential


Roughly 50% of all commercial biologic (therapeutic
protein products) are lyophilized.


Lyophilization

technology and the expertise to use it
are vital to the ability of the biopharmaceutical
industry to prepare and market life
-
saving
injectable

medicines


Lyophilization

is the most challenging (and most
expensive) of all sterile product manufacturing unit
operations


Final Comments

Acknowledgements




Thanks to all my Baxter R&D colleagues for
the great collegiality we have and what
makes my job so enjoyable.


Thanks to the following individuals for
providing some of the slides I showed


Dr. Steven Nail




Ms. Lisa Hardwick


Ms. Wendy Saffell
-
Clemmer


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Baxter Confidential