25 VERISEQ Nucleation

TLAA Linde ProTech Training 200

Chemistry & Energy / Pharma Fine & Spec
Steven Finley
Shanghai, China The Linde Applications Academy
July 2013 The Linde Applications Academy 1
 Key concepts

What is nucleation?
The process by which ice crystals (seed crystals) are formed from originating sites
called nuclei

How does it occur naturally?

Suspended particles and/or container defects create nucleation sites

What factors influence nucleation in Bio-Pharma?

In pure systems nuclei form from water, but only after significant super cooling

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What Factors Impact Nucleation?

What is supercooling?
The cooling of a liquid below its freezing temperature without the formation of the solid
phase

How does supercooling affect ice crystals?

Supercooling defines the minimum size of an ice crystal; the greater the supercooling,
the smaller the ice crystal

Why is this important?

The ice crystals formed in freezing create the pathways for vapour flow in subsequent
steps

Name some strategies for controlling supercooling?

Prolonged soaking, vial etching, annealing, etc.

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Nucleation vs. Solidification

What is solidification?

The fusion of ice from liquid into solid form

How does solidification differ from nucleation?

Solidification occurs after nucleation, when the crystal size is already fixed

How do Bio-Pharma formulations solidify?

Solvents crystallization similar to water

API and Excipients crystallization or glass formation at colder temperatures

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Nucleation vs. Solidification

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Impact of supercooling in lyophilisation

High vial to vial variability, non-uniform product attributes- broken vials, cracked cakes, etc.
Significant supercooling = smaller ice crystal
Smaller ice crystals = higher resistivity to vapour flow
High resistivity = longer cycles = higher costs!

Problem: Non-uniform ice nucleation due to

supercooling leads to vial-to-vial non-uniformity

Damaged
product

Undesirable product attributes (valued at > 1M/batch), high operating costs

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Thermograph #1

Exercise: Identify Nucleation

Identify Supercooling

Identify Solidification

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Thermograph #2

Exercise: Identify Nucleation

Identify Supercooling

Identify Solidification

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Advantages?

Discuss the advantages of Thermograph #2

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VERISEQ Nucleation
Lindes proprietary technology for induced nucleation

Ability to control nucleation and induce freezing

Eliminates vial-to-vial variability, enhances product attributes
Shortened processing time, reducing operating costs
Enhances control, robustness and scalability of lyophilisation processes

VERISEQ Nucleation
Sanitary/sterilisable design
Easy to scale and integrate with lyophilisers
Can be retrofitted to existing lyophilisers
Lab versions (non-aseptic) also available

General Schematic for aseptic (sterile) operation

Lyophiliser

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Sanitary Design

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Flexible Integration

Two 50 mm ports required

Nitrogen and clean steam provided by house supply

Stand alone control

Full system integration

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 Description of VERISEQ Nucleation

Ice fog is produced and distributed inside lyophilizing chamber at

desired temperature
Ice fog provides seed crystals for product to nucleate and freeze
Product is exposed only to WFI (trace amounts) and sterile nitrogen
Entire process occurs within minutes

Real time image w/ glycine formulations

showing ice fog induced nucleation

Process occurring within lyophiliser

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 Test results- Evidence of uniform vial nucleation

Successful testing of multiple technical approaches with IMA Life - Lindes OEM partner for process
implementation
Successful testing on model compounds for improved product and process attributes

Mannitol natural nucleation Mannitol VERISEQ nucleation

Vertical lines show individual vial nucleation Variability eliminated
High variability and supercooling observed Supercooling minimized

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Test results- Improvement in pore structure due to larger
ice crystal formation

Mannitol SEM after complete lyo cycle

Pore structure natural nucleation Pore structure-VERISEQ Nucleation

- Large pores indicative of reduced
supercooling, shorter cycles

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Structural Comparison

Pore structure with Natural Pore structure with Induced

nucleation nucleation

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Test results- Improved visual appearance

Mannitol final visual Mannitol final visual

appearance natural appearance VERISEQ
nucleation Nucleation

Better cosmetic appearance

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Test results- Improved drying profiles

VERISEQ Nucleation

Mass flow of water indicating speed of drying

Higher mass flow observed with VERISEQ Nucleation indicating faster speeds of drying

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Attribute Comparison

Inspection Results

Batch Broken/ Collapsed/ Other Cycle

Cracked Deformed Cake Duration
Uncontrolled Nucleation 0 0 0 119 Hours

Ice Fog Nucleation 0 0 0 119 Hours

Ice Fog Nucleation with cycle 0 0 0 80 Hours

optimization
Reconstitution Results
Batch Test 1 Test 2 Test 3

Uncontrolled Nucleation 145.03 sec 208.66 sec 130.33 sec

Ice Fog Nucleation 120.47 sec 120.53 sec 128.31 sec

Ice Fog Nucleation with cycle 29.72 sec 37.31 sec 30.61 sec
optimization
Moisture Results Specification = < 5%
Batch Natural Nucleation Ice Fog Ice Fog Enhanced

Percent Moisture by KF 3.76% 3.96% 3.49%

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Summary

VERISEQ Nucleation, Lindes proprietary technology for induced nucleation, can be

utilized on laboratory, pilot, and production scale freeze dryers

Induced nucleation/solidification results in larger and more consistent crystal

structures than natural nucleation allows

Product attributes are significantly improved

Drying rate is increased leading to shorter operating cycles

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 Product Service Offering One-Pager
VERISEQ Nucleation

- Develop Ice fog nucleation technology with sterile LIN to achieve

freezing uniformity during lyophilization (freeze drying)
Project goal

- Develop both non-aseptic (lab) system for customer trials and

aseptic (commercial) system for mass-production applications
- Establish strategic partnerships w/ leading OEM(s) to penetrate
high growth pharma market
Customer Benefits:
Increase product yield and process efficiency- savings worth
millions to pharma drug manufacturer
Lyophilizing chamber

- Non-aseptic (lab) system constructed and is Sales and Marketing plan (key points):

Commercialization plan
being tested in preparation for customer trials Individual and/or joint marketing of the System with OEM
- Aseptic (commercial) system prototype has (Commercial Agreement signed);
Project status

been designed, built and successfully tested.
Test results are being used to design and build
an actual aseptic system for customer trials and
certification
- Strategic partnership w/OEM (IMA Life North
America) has been established and commercial
agreement signed
- Gate 4 pending (Q1-2014)
Project & Global marketing (conferences, workshops)
All Systems sales to be made through OEM
OEM to exclusively use and offer to customers the Linde
Technology for controlled nucleation in freeze drying
Linde to exclusively supply the Linde Technology to OEM in
the field of pharmaceutical freeze drying
Market Potential: 14.2M (Cumulative sales)
NPV: 4.7 M

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 Questions?

Contact:
Steven Finley
Head of Pharma, Fine and Spec

Linde Gases Division, 575 Mountain Avenue,

Murray Hill, NJ 07974, USA

+1 908-771-1961
steve.finley@linde.com

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Intellectual Property

All training material is internal, protected intellectual property of Linde AG.

Do not forward, send or hand out information to unauthorized people!

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Thank you very much

for your attention!

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