Evaluation of Container Closure Systems

at Low Temperatures

Alejandra Nieto-Pena, Helen Brown,

Holger Röhl Pharmaceutical Development & Supplies, PTD Biologics Europe (PTDE-P)

Frozen storage of Drug Product

At early stages of development frozen Drug Products are sometimes utilized

Some commercial products (e.g. vaccines) are also stored frozen:

VARIVAX

HZV (Herpes zoster vaccine)

MMR(V) (Masern, Mumps and Röteln; (Windpocken /Varizellen))

Storage conditions: -50°C to -15°C

Note: Shipment of frozen product is often performed using dry ice (-80°C)

Considerations in order to ensure Quality & Safety of a Frozen Drug Product include

Stability upon freezing/thawing

Process variability & consistency of product freezing and thawing

Process availability & reproducability of thawing product (e.g. on clinical site).

Ability/capability to store and handle frozen product in a Clinical Hospital or Study center

As handling (thawing) is performed prior administration, what is the ability to inspect product prior use for e.g. particulates, turbidity or any other liability from the freezing/thawing process?

Impact of low temperatures on the Container Closure System (CCS) = Impact on Container Closure Integrity (CCI)? Because glass transition temperature Tg of standard stoppers is in the range of -50°C to -65°C

Is CCI still guaranteed during storage and transportation?

Why should we care about CCI at low temperatures?

EP: “In all cases the container and closure are required to maintain the sterility of the product throughout its shelf life.”

Any breach in the integrity could lead to a serious risk to the product and to the patients or healthy volunteers.

Could e.g. lead to

Compromise of sterility / microbiological quality

e.g.

Microbiological ingress

pH changes

Increased API oxidation

Increased level / new types of impurities/degradants Potency changes

CO2 ingress (dry ice shipment)

Oxygen ingress

Loss of vacuum / Overpressure

How can we assess CCI for a frozen Drug Product?

• Sterility testing itself cannot replace CCI testing

• CCI testing should use the «frozen» product, not the product after thawing, given that the thawing procedure may «re-seal» of leak that existed in the frozen state.

• The ensurance of CCI needs to consider variability of processes, packaging materials, probabilistic nature of leaks and microbiological ingress

• Microbiologal ingress (mCCI) testing would not work with a frozen product

• Current ideas to test CCI for a frozen Product

– Laser-based Headspace Analysis?

– He based Analysis?

Recent publications:

Mainly based upon Headspace (Lighthouse)

Laser-based Headspace Analysis

• Based upon frequency modulated spectroscopy (FMS)

• Measures changes in internal headspace: O2, CO2, water vapor,

pressure

• Non-destructive

• Rapid (short measurement times)

• Sensitivity

But

• Indirect (no direct assessment of leak)

• Not sufficiently sensitive = Waiting time (days to weeks, depending

upon leak size)

• Need for preparing reference samples

• Lack of correlation to mCCI

6

Recent innovation:

“West” method to directly assess CCI at low

temperatures

• He MS Direct measurement

• Non-destructive

• No need to remove product

• Sensitivity

But

• Total measurement time per sample ? Diffusion controlled

measurements / time to reach equilibrium?

• For liquid filled vials: He absorption into liquid?

• If He diffuses out of the vials this will slow down diffusion

• Lack of correlation to mCCI

“Testing Closure Integrity at Low Temperatures".

P. Cummings, B. Jacobs. West Pharmaceutical. Web: contract pharma on 1.Apr.2014

http://www.contractpharma.com/issues/2014-04-01/view_features/testing-closure-integrity-at-low-temperatures/

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Frozen Product CCI… own development

• We developed a technology to evaluate pCCI in

frozen products ourselves (ThermCCI)

– Highest sensitivity (used pCCI He leakge

testing with excellent sensitivity)

– Direct measurement

– He leak: Well established method

– Quick

– Can adjust temperature (and test ramps)

– Correlation to mCCI already established,

acceptance criteria available

Comparison of microbial ingress vs flow rate

in pCCI/He leakage: deriving acc.crit. for pCCI

- Artificial leaks by copper wire

- Different CCS configurations

Microbial ingress occurred for most samples for leak rates >10 -5 mbar*L/s

(comparable to Kirsch et al) but for one sample even at ca 5x 10 -6

=> Acceptance criteria = definition of «CCI tightness»

for He leakage (pCCI) set to «< 1.0 x 10 -7 mbar*L/s»

Microbial ingress

Experimental Set up – Evolution He-leak test method

Thermo Chamber

“Passive Cooling”

Thermo Chamber

“Active Cooling”

2014 (1st Generation) 2015 (2nd Generation) 2004

Testing Device

“Room Temperature”

10

ThermCCI “Passive Cooling” @ -20 °C

THERMO CHAMBER WORKS Target temperature is maintained at the

surface in contact with vial for ca. 8 hours

Custom-made adapters

for vial neck size (13 & 20

mm)

Temperature monitoring

of both brine solution

and vial neck area Brine (r.t.) is pre-

conditioned @ -80

°C

Frozen

salt

solution

-20±5 °C

Thermometer 1

Vial Temp (°C)

Thermometer 2

Bath Temp (°C)

8 hours

Tem

pera

ture

, °C

Time, h Confidential 11

Vial Temp. -63±5 °C

ThermCCI Active Cooling below -20 °C

DIRECT He-leak test @-59.5

°C

O-ring area at -60 °C (left)

and at + 25 °C (right)

Cooling fluid inside

Thermo Chamber

Chiller Temp. -67±5 °C Confidential

THERMO CHAMBER WORKS Target temperature is maintained at the

surface in contact with vial for as long as desired

-60 °C +25 °C

Inlet

Outlet

System is tight

Therm CCI: Feasibility Study

Impact of … on CCI of

frozen vials

- Storage time

- Rubber stopper

composition

- Artificial leaks

Sample preparation

- 2 mL vials (stoppered and crimped) filled with He (no

liquid)

- Two rubber formulations :

A (Tg = -66.5°C) & B (Tg = -105°C)

- Storage for 1 week at RT and -75°C

- 10 vials per configuration

- Artificial leaks with Cu wires (∅ = 20 µm and 60 µm)

CO2

Headspace

Analytics

He-leak tests

@ RT

@ -20±5°C

@ -60±5°C CT Scans

Therm CCI: Experimental runs (Stopper A)

Storage

conditions

Copper wire

diameter (mm)

(artificial leak

creation)

Storage

time

(week)

CO2

Headspace

% Leaking

vials

He-leak

@RT

% Leaking

vials

He-leak

@-20 °C

% Leaking

vials

He-leak

@-60 °C

% Leaking

vials

RT

no wire 1 0 0 0 0

20 1 0 0 0 0

60 1 30 * 100 100 100

Dry ice

no wire 1 0 0 0 0

20 1 20 (11) 20 20

60 1 100 100 100 100

Headspace Analysis : Vials are considered tight if P (CO2) <0.07 % atm

He-leak test: Vials are considered tight if leak rate is <1*10-7 mbar*L/sec

* Vials stored in same room than dry ice samples; ingress of CO2 in ambient air? Values measured are in the range 0f

0.07-0.10%

Therm CCI: Experimental runs (Stopper A)

Storage

conditions

Copper wire

diameter (mm)

(artificial leak

creation)

Storage

time

(week)

CO2

Headspace

% Leaking

vials

He-leak

@RT

% Leaking

vials

He-leak

@-20 °C

% Leaking

vials

He-leak

@-60 °C

% Leaking

vials

RT

no wire 1 0 0 0 0

20 1 0 0 0 0

60 1 30 * 100 100 100

Dry ice

no wire 1 0 0 0 0

20 1 20 (11) 20 20

60 1 100 100 100 100

CONCLUSIONS

The Therm CCI He method was more sensitive than the CO2 Headspace method.

At Drug Product Storage at dry ice, increasing number of frozen vials showed leakage

also with a 20 µm wire diameter, sugesting these vials are not sufficiently tight any

more.

Frozen vials exhibit (expected) leakage with copper wires artifical leaks (60 µm wire).

Vials without inserted Cu wires are tight stored at 60°C under static conditions.

Therm CCI: Experimental runs (Stopper B)

Storage

conditions

Copper wire

diameter (mm)

(artificial leak

creation)

Storage

time

(week)

CO2

Headspace

% Leaking

vials

He-leak

@RT

% Leaking

vials

He-leak

@-20 °C

% Leaking

vials

He-leak

@-60 °C

% Leaking

vials

RT

no wire 1 0 0 0 100

20 1 0 0 100 100

60 1 30* 100 100 100

Dry ice

no wire 1 10 0 0 100

20 1 100** 33 100 100

60 1 100*** 100 100 100

Headspace Analysis : Vials are considered tight if P (CO2) <0.07 % atm

He-leak test: Vials are considered tight if leak rate is <1*10-7 mbar*L/sec

** All vials with 20µm wires exhibit overpressure (> 1 atm)

*** All vials with 60µm wires do not exhibit overpressure

Therm CCI: Experimental runs (Stopper B)

Storage

conditions

Copper wire

diameter (mm)

(artificial leak

creation)

Storage

time

(week)

CO2

Headspace

% Leaking

vials

He-leak

@RT

% Leaking

vials

He-leak

@-20 °C

% Leaking

vials

He-leak

@-60 °C

% Leaking

vials

RT

no wire 1 0 0 0 100

20 1 0 0 100 100

60 1 30* 100 100 100

Dry ice

no wire 1 10 0 0 100

20 1 100** 33 100 100

60 1 100*** 100 100 100

CONCLUSIONS

The Therm CCI He method was more sensitive than the CO2 Headspace method.

Frozen vials showed leakage

Decreasing temperature is leading to an increase of the number of leaky vials even

with no artificial leaks or with an inserted Cu wire of 20 µm

Results are an indicator for poor fit and/or crimping of the CCS => unsuitable CCS

Stopper B Stopper A

CT scans of vials crimped with stopper A and

B

CONCLUSIONS

CT scans reveal reveal differences between vials with stopper A and B which cannot be

detected by visible inspection

Potential root cause for differences in CCI behavior

Overall conclusions

ThermCCI: Feasibility studies

• ThermCCI set up is able to measure CCI via He-leak test

method directly at target temperature (down to -60 C)

• Very sensitive method, and more sensitive and suitable to

Headspace analysis

• If leaks are present the impact on CCI seems to be higher

at lower storage temperatures

Therefore, selection of stopper is very important

(fitting)!

• Both CCS tested did not fulfill CCI requirements when

stored frozen, thus, frozen DP storage/shipment of these

CCS is not suggested

Outlook

• Broaden data base

• Include more rubber stopper formulations into studies

• FE calculations to assess artificial leak sizes induced by

Cu wires with varying diameter

• Additional investigations of effect of

• Capping

• Shipping stress /Drop test

• Freeze/Thaw cycles

• Improvement of system (to reach temperatures below -

70°C)

• Publication of work

Acknowledgements

Hanns-Christian Mahler

Michael Adler

Miriam Printz

Monica Gisin

Jonas Nikoloff

Sascha Dreher

Yutaka Fukazawa (Daikyo Seiko)

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