1 Baxter Confidential Principles and Applications of Lyophilization to Biotechnology Michael J. Akers, Ph.D. Baxter BioPharma Solutions Bloomington, Indiana

1Baxter Confidential

Principles and Applications of Lyophilization to Biotechnology

Michael J. Akers, Ph.D. Baxter BioPharma

SolutionsBloomington, Indiana

BIOMAN 2010 ConferenceIvy Tech Community

CollegeBloomington, IndianaJuly 13, 2010

Organization of Presentation



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


Focus of Bloomington R&D Formulation Development of Small Volume Injectables

Liquid Lyophilized Extensive experience with

ProteinsMonoclonal AntibodiesSmall 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


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

Hea

t F

low

, W/g

Endotherm Up

-30 -28 -26 -24 -22 -20 -18 -16

2 W/g

20 W/g

Detectors

to

VacuumPump

Condenser~ - 80°C

Chamber

Front Door

TestSection

1

2

Vacuum

H-3236


Baxter BioPharma SolutionsPharmaceutical Research and DevelopmentBloomington, IN

Top: Tim Paul, Lisa Hardwick, Nathan Pease, Wendy Saffell-Clemmer, Larry CallahanMiddle: Rhonda Haley,Wei Kuu, Melissa Beard, Mike Hildreth, Karen Abram, Kevin O’Bryan, Elizabeth Oslos, Michael AkersBottom: Lindsay Stanford, Angie Kruszynski, Nathaniel Fair, Jackie Karty, Greg Sacha, Hoang Mitchell, Dan Staples, Kelly Roby, Steven Nail

Baxter Confidential 8

Market segment by therapeutic area and molecule class

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


9

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


Best Known (Best Selling) Lyophilized Biologics (Biopharmaceuticals)

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%).



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 solutionSterile 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% inspectionStorage at appropriate

temperature (usually 2-8ºC)

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


Today’s State of the Art Technology UsesAutomated Systems To Transfer ProductFrom Production Line Into the Freeze-Dryer

14

Freeze-Dryer Subsystems (What Must Be Operating Smoothly and Maintained for Lyophilization Process To Succeed?)


Baxter Pharmaceutical Solutions

• Heat Transfer System• Refrigeration System• Vacuum System• Stopper-in-Place System• Clean-in-Place System• Control System• Loading/Unloading System

• 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 $


Courtesy of Samir U. Sane, Ph.D., Genentech, Inc.


Product temperature lower than shelf

temperature during primary drying

because energy is consumed as ice

sublimes to vapor.


Flow of Vapor from Chamber to Condenser

T

Connecting Duct

Shelves with

Vials

Vacuum

Pump

Door

Water Vapor

Flow

T P

Product

ChamberCondenser

Chamber

Mushroom Valve

(open position)

Condenser Coils


Thermal Fluid Shelf

Frozen Solution

Sublimation Front

Dry Cake

Heat

Tra

nsfe

r

Mass T

ran

sfe

r

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


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


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


Common Lyophilized Products• Pharmaceuticals – large and small

molecules• Bacteria• Viruses• Vaccines• Plasma• Small Zoological Specimens (Taxidermy)• Fruit• Coffee• Flowers • Water-Damaged Documents


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 cycles24


Baxter Pharmaceutical Solutions

Challenges in Lyophilization of Protein Pharmaceuticals


• 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 –30oC. 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)

Te

mp

era

ture

(°C

)

0

50

100

150

200

250

300

Pre

ss

ure

(m

To

rr)

Thermocouples


• 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
