through vial impedance spectroscopy (tvis)€¦ · global pharmaceutical market 2015 and 2021...

Through Vial Impedance Spectroscopy

Through Vial Impedance Spectroscopy (TVIS)

A new method for the development of manufacturing processes for injectable drug product

Prof. Geoff Smith

10th International Workshop on Impedance Spectroscopy 28-29th September 2017, Chemnitz, Germany

2 Through Vial Impedance Spectroscopy

Pharmaceuticals (From drugs molecules to products)

Man-made drugs – small molecules (chemical synthesis) to large molecules (biotechnology)

Formulation development • Drug products (i.e. dosage form: tablets, injections) etc. • Healthcare and cosmetics product (i.e. nutrition)

Production

Product available in the market • Quality: Safety & Efficacy

Aspirin 21 atoms

Hormone ~3000 atoms

Antibody 25,000 atoms

QC Pharmacopoeial tests


Biopharmaceutical in Market from 1982-2014 (classified by therapeutic categories)

Antihemophilic factor, human recombinant

0

5

10

15

20

25

Blood factors Other bloodrelated

Hormones Growthfactors

IFN,IL &TNF Vaccines mAbs Other

Pro

du

ct a

pp

rova

l (%

) Hemoglobin

Growth hormone

GM-CSF

Interferon alfa

Varicella-zoster virus

Nivolumab


Global Pharmaceutical Market 2015 and 2021

Newrzella A. (2017) Pharma & Biotech 2017 – Review of Outsourced Manufacturing

0

200

400

600

800

1000

1200

1400

2015 2021

Mar

ket

size

/ $

bn

Year

Biosimilars

Biologics

OTC

Generics

Patented/Originatorsmal molecules

The biologics market increases rapidly from 16.6% in 2015 to 22.2% in 2021.


Monoclonal antibodies (mAbs) • A monospecific immunoglobulin

• Medicinal application of mAbs

o Diagnostic application (i.e. immunoassay, immunoscintigraphy), e.g. Prof.Abdelhamid

o Therapeutic applications (i.e. Cancer, Transplantation, Immune disease etc.)

Antibodies bind with

antigens on the surface of

target cells

T-cell bind with

antibodies

Lead to cell lysis or phagocytosis

target cells

are destroyed

Example of mAbs mechanism of action

6

Monoclonal antibodies (mAb)

Oncology disease

Immune disorder

Leukaemia

Breast cancer

Lung cancer

Colorectal cancer

Organ Transplantation

Crohn’s disease

Multiple Sclerosis

Rheumatoid arthritis

Psoriasis

7

Drug Product Development

http://phrma-docs.phrma.org/sites/default/files/pdf/biopharmaceutical-industry-profile.pdf

Freeze Drying Process

9

Advantages of Lyophilization

• Lyophilization commonly used for

o Large Molecule Drugs (e.g. proteins, DNA)

o Small Molecules Drugs (e.g. penicillin)

o Microorganisms (e.g. bacteria, virusus)

o Blood products

Lyophilization process Dry Powder

Increase Stability

Easy to Dissolving

Easy to transport Low moisture content

More surface area & porous

Low temperature

process

Zoster vaccine (Zostavax®) Azithromycin injection.

(Zithromax®)

https://www.pharmpro.com/article/2017/03/lyophilization-basics

“40% of biologically based products have to be freeze dried” “80% of the available products lyophilized in vial”

http://www.genengnews.com/gen-articles/lyophilization-growing-with-biotechnology/1083


Limitation of Lyophilization Technology

• Complicate

• Costly

• Long process

• Difficult to scale up

• Variation between batch


Lyophillization or Freeze Drying Process

• A technique which dries product at low temperature through sublimation process

• It consists of three main steps : Freezing, Primary drying and Secondary drying

Pre

ssu

re /

mb

ar

Tem

per

atu

re /

oC

Time / h

Product Temperature

Shelf Temperature

pressure

Freezing Annealing (option)

Primary Drying

Secondary Drying

Freeze Dryer Shelf

Equilibrium product temperature of all vials

𝑇𝑚





Pre

ssu

re /

mb

ar

Tem

per

atu

re /

oC

Time / h

Product Temperature

Shelf Temperature

pressure


Primary Drying

Secondary Drying

𝑇𝑚

Freeze Dryer Shelf

Temperature of the product (liquid state) decreases (Tp) as shelf temperature decreases (Ts)





Pre

ssu

re /

mb

ar

Tem

per

atu

re /

oC

Time / h

Product Temperature

Shelf Temperature

pressure


Primary Drying

Secondary Drying

𝑇𝑚

Liquid product supercools below the melting point: Melting temperatures of ice in frozen solution would be less than that of pure water, owing to the

freezing point depression of the solutes

Freeze Dryer Shelf


Pre

ssu

re /

mb

ar

Tem

per

atu

re /

oC

Time / h

Product Temperature

Shelf Temperature

pressure


Primary Drying

Secondary Drying




Ice crystal growth from the bottom of the vial (typically takes less than 2 min).

Release of heat causes a spike in the product temperature

Freeze Dryer Shelf

Ice nucleation





Pre

ssu

re /

mb

ar

Tem

per

atu

re /

oC

Time / h

Product Temperature

Shelf Temperature

pressure


Primary Drying

Secondary Drying

Temperature hold to ensure complete solidification of ice

Freeze Dryer Shelf

Ice layer

Temperature of frozen solid (Tp) now in equilibrium with the shelf

but Tp > Ts by 2-3 oC





Pre

ssu

re /

mb

ar

Tem

per

atu

re /

oC

Time / h

Product Temperature

Shelf Temperature

pressure


Primary Drying

Secondary Drying

Crystal growth during annealing

Freeze Dryer Shelf

Ice layer





Pre

ssu

re /

mb

ar

Tem

per

atu

re /

oC

Time / h

Product Temperature

Shelf Temperature

pressure


Primary Drying

Secondary Drying

Freeze Dryer Shelf

Ice layer





Pre

ssu

re /

mb

ar

Tem

per

atu

re /

oC

Time / h

Product Temperature

Shelf Temperature

pressure


Primary Drying

Secondary Drying

Increasing shelf temperature (ramp), increases ice temperature and partial

pressure until 𝑃𝑖𝑐𝑒 = 𝑃𝑐 and drying (sublimation) starts

Freeze Dryer Shelf

Radiation

Convection

Direct

conduction

Convection

Ice layer

𝑃𝑖𝑐𝑒 = 𝑃𝑐





Pre

ssu

re /

mb

ar

Tem

per

atu

re /

oC

Time / h

Product Temperature

Shelf Temperature

pressure


Primary Drying

Secondary Drying

Once the ice is removed then self cooling stops and the product

temperature can now catch up with the shelf temperature.

When 𝑇𝑃 ≥ 𝑇𝑠; then drying is assumed to have stopped

Freeze Dryer Shelf

Dried layer





Pre

ssu

re /

mb

ar

Tem

per

atu

re /

oC

Time / h

Product Temperature

Shelf Temperature

pressure


Primary Drying

Secondary Drying

Freeze Dryer Shelf

Dried layer


Process Analytical Technologies

Challenging in development and manufacture of freeze-dried

biopharmaceuticals

Regulatory requirements

Characteristic of protein

therapeutic (i.e. unstable)

Process variation

(can affect productivity,

consistency & repeatability)

Process Analytical Technology (PAT)

Definition by US FDA:

A mechanism to design, analyze and control pharmaceutical manufacturing process through the measurement of Critical Process Parameters (CPP) which affect Critical Quality Attributes (CQA)).

- Manometric Temperature Measurement (MTM) - Tunable Diode Laser Absorption Spectroscopy (TDLAS) Limitation : - Batch method (representative parameter) → not

suitable for high variation batch (e.g. edge effect) - TDLAS is difficult to calibrate and costly


Introduction to the TVIS System • Impedance spectroscopy characterizes the ability of materials to conduct

electricity under an applied an oscillating voltage (of varying frequency)

• Impedance measurements across a vial rather than within the vial

• Hence “Through Vial Impedance Spectroscopy”

• Features

• Single vial “non-product invasive”

• Both freezing and drying characterised in a single technique

• Non-perturbing to the packing of vials

• Stopper mechanism unaffected

SV product temperature

SV sublimation rate

SV end point


Through Vial Impedance Spectroscopy (TVIS) Introduction


Freeze drying chamber

Stimulating voltage

Resultant current

LyoViewTM analysis software

LyoDEATM measurement software

Junction box

TVIS system (I to V convertor)

Pass through

TVIS measurement vial


Impedance Analyzer for Lyophilization Process

• Through Vial Multi Channel Impedance Analyzer

• Impedance measurement specially optimized for lyophilization experiments (contact method)

• Five sequentially measuring impedance channels

• All five channels share a common excitation signal

• Automatic voltage excitation amplitude adjustment

• Current Gain 109 (1 Gigaohm trans impedance amplifier gain)

• Five synchronized type K thermocouple measuring ports


Equivalent electrical circuit model

• An equivalent electrical circuit model is created by combining the circuit elements which includes the solution resistance (𝑅𝑠) and the capacitances of the glass-solution interface (𝐶𝐺) and the solution (𝐶𝑠) in an appropriate configuration of series and parallel elements.

CG is the capacitance of the glass-solution interface, CS and RS are the capacitance and resistance of the solution

𝑹s

𝑪G 𝑪s


Impedance to Complex Capacitance • The impedance of the model can be calculated from the following equation

𝑍∗Total = 𝑍∗ 𝐶G + [

1

𝑍∗ 𝑅S

+ 1

𝑍∗ 𝐶S

]

𝑍∗Total =

1

𝑖𝜔𝐶𝐺+

𝑅𝑆

1 + 𝑖𝜔𝑅𝑆𝐶𝑆

which re-arranges to

𝑍∗Total =

1 + 𝑖𝜔𝑅𝑆(𝐶𝐺 + 𝐶𝑆)

𝑖𝜔𝐶𝐺 + 𝑖𝜔2𝑅𝑆𝐶𝐺𝐶𝑆

• Impedance can be expressed in terms of a complex capacitance

𝐶∗Total =

1

𝑖𝜔𝑍∗Total

=𝐶𝐺 + 𝑖𝜔𝑅𝑆𝐶𝐺𝐶𝑆

1 + 𝑖𝜔𝑅𝑆 𝐶𝐺 + 𝐶𝑆

• The complex capacitance can also be expressed in form of real part and imaginary part

𝐶∗ = 𝐶′ + 𝑖𝐶″

• From the complex capacitance formula, the expressions for real and imaginary capacitance can be calculated to explain the origin of interfacial polarization peak. This achieved by multiplying the nominator and denominator by the complex conjugate of the denominator and by grouping the real (𝐶′) and imaginary (𝐶″) parts

𝐶′ =𝐶𝐺+𝜔2𝑅𝑆

2𝐶𝐺𝐶𝑆 𝐶𝑆+𝐶𝐺

1+ 𝜔𝑅𝑆 (𝐶𝑆+𝐶𝐺2 and 𝐶″= −

𝜔𝑅𝑆𝐶𝐺2

1+(𝜔𝑅𝑆((𝐶𝑆+𝐶𝐺))2


Dielectric loss spectrum of frozen water at -27 oC

• A frequency of

• If 𝐶𝐺 > 𝐶𝑆 then

• Which explains the sensitivity of 𝐶″𝑃𝐸𝐴𝐾 to

the height of the ice layer

𝐶″𝑃𝐸𝐴𝐾 =𝐶𝐺

2

2(𝐶𝑆+𝐶𝐺)

𝐹𝑃𝐸𝐴𝐾 =1

2𝜋𝑅S (𝐶𝑆 + 𝐶𝐺)

𝐶″𝑃𝐸𝐴𝐾 ≅ 𝐶𝐺

𝑅s

𝐶G 𝐶s

0.00

0.10

0.20

0.30

1 2 3 4 5 6

-C″

/pF

Log Frequency

𝐶″𝑃𝐸𝐴𝐾 = 0.294 𝑝𝐹

𝐹𝑃𝐸𝐴𝐾 = 3.08

0.618 pF 0.031 pF

2.02 x 108 Ω


Imaginary Part of Capacitance Real Part of Capacitance

High frequency

Liquid state Liquid state Frozen solid

Frozen solid

low frequency

Annealing = Re-heating and Re-cooling

Re-heating Re-heating

Intermediate frequency

Re-cooling

Re-cooling

low frequency

Primary drying Primary drying

low frequency

TVIS Response Surface (3D-Plot)


Phase Separation in freezing step 5%w/v Lactose solution (frozen)

0.0

0.1

0.2

0.3

0.4

0.5

1 2 3 4 5 6

-C″

/ p

F

Log Frequency

Water (frozen)

0.0

0.2

0.4

0.6

0.8

1.0

1 2 3 4 5 6

-C″

/ p

F

Log Frequency

ice Unfrozen Fraction

Add the equivalent circuit here Add the equivalent circuit here

Ice peak

Ice peak

Unfrozen fraction peak


Impedance and Capacitance Spectrum

5%w/v Lactose solution

+ 20.3oC

0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

1 2 3 4 5 6

-C″

/pF

C′ /

pF

Log Frequency

-90-80-70-60-50-40-30-20-100

2

4

6

8

10

12

1 2 3 4 5 6

Thet

a

Log

|Z|

Log Frequency

Liquid state



0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

1 2 3 4 5 6

-C″

/pF

C′ /

pF

Log Frequency

-90-80-70-60-50-40-30-20-100

2

4

6

8

10

12

1 2 3 4 5 6

Thet

a

Log

|Z|

Log Frequency

Solid (frozen state) lower temp


-30.4 oC



0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

1 2 3 4 5 6

-C″

/pF

C′ /

pF

Log Frequency

-90-80-70-60-50-40-30-20-100

2

4

6

8

10

12

1 2 3 4 5 6

Thet

a

Log

|Z|

Log Frequency

Solid (frozen state) high temp


-20.4 oC



0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

1 2 3 4 5 6

-C″

/pF

C′ /

pF

Log Frequency

-90-80-70-60-50-40-30-20-100

2

4

6

8

10

12

1 2 3 4 5 6

Thet

a

Log

|Z|

Log Frequency

Dry state


-10.8 oC


Through Vial Impedance Spectroscopy (TVIS)

• TVIS measurement relate to both the electrical resistance and electrical capacitance of the vial contents.

0.0

0.2

0.4

0.6

0.8

1 2 3 4 5 6

- C

″/ p

F

Log Frequency

FPEAK FPEAK

Liquid

State

Solid

State

0.00.10.20.30.40.50.6

1 2 3 4 5 6

-C

″ /

pF

Log Frequency

C″PEAK

incr

ease

Drying time

0.0

0.2

0.4

0.6

0.8

1 2 3 4 5 6

-C″

/pF

Log Frequency

-38 oC

-18 oC

Monitoring Phase Behaviour (ice nucleation temperature and solidification end points

FPEAK temperature calibration for predicting temperature of the product in primary drying

Drying rate surrogate (from dC″PEAK /dt )

C′ (real part of the complex capacitance) is highly sensitive to low ice volumes; therefore

it could be used for determination end point of primary drying


0.0

0.2

0.4

0.6

0.8

1.0

1 2 3 4 5 6

C″/

pF

Log Frequency

• FPEAK profile during annealing has ‘similar’ profile with product temperature.

• Assuming thermal equivalence between the thermocouple (TC) vial and TVIS vial, then the temperature calibration from annealing might be employed for the prediction of temperature during primary drying

Temperature Calibration

-40 oC

-60-50-40-30-20-100

2.0

2.5

3.0

3.5

4.0

4.5

5 6 7 8 9

Tem

per

atu

re/o

C

Log

F PEA

K

Time / h

-20 oC

y = -4.7474x2 + 56.64x - 160.16 R² = 0.9998

-50

-40

-30

-20

-10

2.6 2.8 3.0 3.2 3.4 3.6 3.8

Tem

per

atu

re /

oC

Log FPEAK

Shelf Temp. Product Temp.

TVIS

Temperature FPEAK

37

Temperature Prediction in Primary Drying

• Good agreement between product temperature (by TC) and T(FPEAK)

-50

-40

-30

-20

-10

0

12 13 14 15 16

Tem

per

atu

re /

oC

Time / h

• Temperature calibration curve selected for temperature prediction in primary drying : T(FPEAK)

y = -4.7474x2 + 56.64x - 160.16 R² = 0.9998

-50

-40

-30

-20

-10

2.6 2.8 3.0 3.2 3.4 3.6 3.8

Tem

per

atu

re /

oC

Log FPEAK

Shelf Temperature (𝑇𝑆)

Thermocouple T(FPEAK)

Before drying

during drying

TC T(FPEAK)

𝑇(𝐹𝑃𝐸𝐴𝐾) > 𝑇𝑠

Before drying

𝑑𝑄

𝑑𝑡

TC T(FPEAK)

𝑇(𝐹𝑃𝐸𝐴𝐾) < 𝑇𝑠

during drying

𝑑𝑄

𝑑𝑡

𝑑𝑄

𝑑𝑡= 𝐿 ∙

𝑑𝑚

𝑑𝑡


Compensation of C″PEAK by T(FPEAK)

y = -8E-05x2 - 0.0016x + 0.8962 R² = 0.9993

0.80

0.82

0.84

0.86

0.88

0.90

-45 -40 -35 -30 -25 -20

C″ P

EAK

Temperature /oC

0.4

0.5

0.6

0.7

0.8

0.9

1.0

12 14 16 18

C″ P

EAK /

pF

Time / h

Ĉ″PEAK

C″PEAK

13.67 h

-44

-42

-40

-38

-36

-34

-32

-30

12 14 16 18

Tem

per

atu

re /

oC

Time / h

13

.67

h d

ryin

g st

arts

Thermocouple

T(FPEAK) 195 µbar

0.0

0.1

0.2

0.3

0.4

-50 -45 -40 -35 -30 -25

Pre

ssu

re /

mb

ar

Temperature / oC

195 µbar

−36

oC

T(FPEAK) during primary drying is used for compensation


0.0

0.2

0.4

0.6

0.8

1.0

12 14 16 18 20 22 24

C″ P

EAK /

pF

Time / h

• Drying rate (g/h) for Ĉ″PEAK

Drying rate calculation

)()()(

)()(

ˆ)

ˆˆ(

initialPEAKinitialend

endPEAKinitialPEAK

C

regionelectrodewithinmassice

TimeTime

CCrateDrying

152.083.0

69.3)

3.144.17

47.083.0(

hgrateDrying

(0.83 pF, 14.3 h)

(0.47 pF, 17.4 h)


Summary

• Temperature calibration of the TVIS parameter (𝐹𝑃𝐸𝐴𝐾) for ice during an additional temperature cycling stage applied to a prediction of ice temperatures during the initial (few hours) of primary drying

• Temperature compensation of TVIS parameter (𝐶𝑃𝐸𝐴𝐾″ ) allows for an accurate

estimation of ice mass during primary drying as evidenced by comparable results of drying rate between the determined by TVIS and that determined (gravimetrically) by loss weight

Non-invasive real time information for characterising the freeze drying process


Future Work

• Development mapping a drying characteristics from lab scale to production

o Determination of heat transfer coefficients (𝐾𝑉)

o Determination of dry layer resistance (𝑅𝑃) to predict drying efficiency

• Investigation the molecular dynamic of the unfrozen fraction

o Monitoring product stability

o Examine the mechanical strength of the freeze dried product (i.e. collapse behaviour)

• Develop (new) continuous drying technologies


• De Montfort University, School of Pharmacy

o Evgeny Polygalov: co-inventor of TVIS instrument

o Yowwares Jeeraruangrattana. PhD student

o Irina Ermolina. Senior Lecturer

• Sciospec Scientific Instruments

Commercial Development of TVIS instrument

o Martin Bulst

o Sebastien Wegner

Acknowledgements, Recent Projects & Collaborators

Biopharmaceutical Stability at Room Temperature

Analytical Technologies for the Stabilization of Biopharmaceuticals

Government Support for industry

LyoDEA Lyophilization process analytics By dielectric analysis

http://www.nibsc.org/

through vial impedance spectroscopy (tvis)€¦ · global pharmaceutical market 2015 and 2021...

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