Surfactants: Raw Material and Drug Product Control Strategy · Surfactants: Raw Material and Drug Product Control Strategy Atanas Koulov Lonza Drug Product Services NBC | Boston |

1 | P HA RMA & B IOT E CH | P ROT E IN T HE RA P E UT ICS MA NUFA CT URING | MAY 20 – 22 , 2015

Pharma&Biotech

Surfactants:

Raw Material and Drug Product Control Strategy

Atanas Koulov

Lonza Drug Product Services

NBC | Boston | 16.05.2016


Forward-Looking Statements

Certain matters discussed in this presentation may constitute forward-looking

statements. These statements are based on current expectations and estimates

of Lonza Group Ltd, although Lonza Group Ltd can give no assurance that these

expectations and estimates will be achieved. Investors are cautioned that all

forward-looking statements involve risks and uncertainty and are qualified in their

entirety. The actual results may differ materially in the future from the forward-

looking statements included in this presentation due to various factors.

Furthermore, except as otherwise required by law, Lonza Group Ltd disclaims any

intention or obligation to update the statements contained in this presentation.


A good control strategy comprises of

Raw materials testing/ qualification

Product characterization throughout

development

Adherence to GMP

Manufacturing process validation

In-process control

Specifications (release, stability)

Stability testing

Based on ICH Q8 (R2), Q10, Q11


Role of surfactants in protein therapeutics

Non-ionic surfactants likely stabilize proteins by competitive interaction with interfaces

The required concentrations are typically derived from formulation studies with

(artificially) induced interfaces (e.g. shaking & freeze/thaw); thermal studies cannot

demonstrate the need to use surfactants for protein stabilization

In certain cases, interaction and colloidal stabilization of the protein by the surfactant

is also found to occur

0 % 0.005 % 0.01 % 0.02 % 0.05 % PS20

Khan, Mahler, Kishore (2015) EJPB 97, 60-67. ; Kiese, Pappenberger, Friess, Mahler (2008) J Pharm Sci 97, 4347-66.


Surfactants (in general)

Types

Ionic (e.g. SDS, CTAB)

Non-ionic (e.g. polysorbates, n-octyl-b-D-glucopyranoside,

Triton X-100)

Zwitterionic (e.g. CHAPS)

Important properties:

CMC (critical micelle concentration)

HLB (hydrophilic/ lipophilic balance)

Aggregation number

Cloud point

Kraft point

Most of these are not used in

parenteral formulations due to

safety-related limitations or

concerns


Surfactants used in parenteral formulations

Polysorbates Poloxamer

w+x+y+z=20

MWav: 1228; CMC = 0.007% (w/v) (0.06 mM); HLB = 16.7

MWav: 1310; CMC = 0.002% (w/v) (0.013 mM); HLB = 15

Avg mol wt: 8350; CMC = 0.064M; HLB = 29

x= 98, y= 67, z= 98


Polysorbate as a pharmaceutical excipient

PS are complex and heterogeneous

mixtures (synthesis uses some

precursors from natural products)

Manufacturing processes may vary/

change

Fatty acid ester PS20 PS80

Caproic

CH3(CH2)4COOH

≤1% -

Caprylic

CH3(CH2)6COOH

≤10% -

Capric

CH3(CH2)8COOH

≤10% -

Lauric

CH3(CH2)10COOH

40-60% -

Myristic

CH3(CH2)12COOH

14-25% ≤5%

Palmitic

CH3(CH2)14COOH

7-15% ≤16%

Palmitoleic

CH3(CH2)5CH=CH(CH2)7COOH

- ≤8%

Stearic

CH3(CH2)16COOH

≤7% ≤6%

Oleic

CH3(CH2)7CH=CH(CH2)7COOH

≤11% 58-85%

Linoleic

CH3(CH2)3(CH2CH=CH)2(CH2)7COOH

≤3% ≤18%

Linolenic

CH3(CH2CH=CH)3(CH2)7COOH

- ≤4%

European Pharmacopoeia, 01-2005:0426 and 80 01-2005:0428


Polysorbate as a pharmaceutical excipient

Borisov et al., J Pharm Sci, 104(3),1005–1018

PS are complex and heterogeneous

mixtures

PS raw materials recommended

storage – ambient, but, DP typically

stored at 2-8C°


Polysorbates may degrade via different mechanisms

Hydrolytic

Non-enzymatic

Enzymatic?

Oxidative

Bates et al., 1973. J Pharm Pharmacol 25:470–477

Kishore et al., J Pharm Sci, 2011, 100:2, 721-731

LaBrenz, 2014, J Pharm Sci,103:2268–2277

Hall et al., J Pharm Sci. 2016 ,105(5):1633-42

Dixit et al., J Pharm Sci. 2016,105(5):1657-66

Donbrow et al., 1978, J Pharm Sci 67:1676–1681

Borisov et al., J Pharm Sci, 104(3),1005–1018;

Porter et al., 1995, Lipids 30: 277–290;

Yin and Porter, 2005, Antioxid Redox Signal 7:170–184;

Kerwin BA 2008. J Pharm Sci 97(8):2924-2935

Kishore et al., J Pharm Sci, 2011, 100:2, 721-731

10 | P HA RMA & B IOT E CH | P ROT E IN T HE RA P E UT ICS MA NUFA CT URIN G | MAY 20 – 22 , 2015

Mechanisms of PS degradation

Oxidation

Auto oxidation via radical mechanism

Initiation (hydrogen abstraction produces free radicals)

Formation of peroxy radicals (reaction with molecular oxigen)

Propagation – intra- or intermolecular hydrogen abstraction

Light or transition metals may accelerate these reactions

Temperature dependent, though significant degradation can

happen at 2-8°C as well

Can happen in formulation (during storage)!

Can happen in placebo

Likely concomitant protein oxidation

May be mitigated by antioxidants, radical scavenger or chelating

agents in the formulation(pat. # WO 2011089062 A2)


Mechanisms of PS degradation

Hydrolysis

Polysorbates (esters) are susceptible to hydrolysis (proteins have

aqueous formulations)

However, reaction seemingly slow under protein DP storage conditions

(2-8°C)

Enzyme-catalyzed hydrolysis

Assumed by esterase (<ppm) that is carried over (HCP)

If enzyme-based, should have pH and temperature optima

Data for enzyme-mediated PS hydrolysis are limited

Large variety of degradation products Kishore et al., Pharm Res, 2011, 28:1194–1210


Analytical methods for characterization of polysorbates

A number of analytical tools are available for PS quantification & PS

degradant characterization:

RP-HPLC/ mixed mode-HPLC assays

(often used in conjunction with FMA)

Detection:

MS

ELSD

CAD

Various gradients depending on the purpose

PDAM assay

Fluorescence micelle assays in various formats

Microspectroscopic methods

FTIR

Raman

GC-MS


Analytical methods for characterization of polysorbates

Availability of a sophisticated analytical toolbox is

key to understanding surfactant degradation:

Raw material testing

Monitoring of surfactant content and stability in formulations

Understanding of degradation pathways – degradation products

are diagnostic of the root cause(s)


Content assays

Fluorescence micelle assay

depends on micelle formation

HPLC or e.g. plate reader format

HPLC assays

RP or mixed mode

CAD, ELSD or MS detection

non-linear calibration curves; different LOD

NMR

Different measurements – different

results!

All content assays are impacted differently by

surfactant degradation

Kishore et al., J Pharm Sci 2011, 100: 2

Hvattum et al. J Pharm Biomed Anal 2012, 62 7– 16


Composition assays

Degradation products of polysorbates

Borisov et al., J Pharm Sci, 104(3),1005–1018 Hvattum et al. J Pharm Biomed Anal 2012, 62 7– 16

Li et al., Anal. Chem. 2014, 86, 5150−5157Siska et al., 2015, J Pharm Sci, 104:447–456


Methods to characterize PS degradation products

Polysorbate degradation products – e.g. particles (FFAs and FA

esters)

FTIR microspectroscopy Raman microspectroscopyCao et al., J Pharm Sci , 2015, 104:433–446 Saggu et al, Pharm Res, 2015,32:2877–2888





development

Adherence to GMP


In-process control


Stability testing


PS are complex and heterogeneous mixtures

POE

POE sorbitan

di- and tri-esters

Isosorbide polyethoxylates

(IPE)

Hewitt D et al., J Chromatogr A. 2011


Importance of raw material testing and storage

Batch-to-batch and supplier variability

Impact of impurities present in the raw material

Free fatty acids present in PS – presumably unreacted starting material

(Siska et al., 2015, J Pharm Sci, 104:447–456)

Presence of peroxides in PS

Singh et al., AAPS PharmSciTech, Vol. 13, No. 2, June 2012

Wasylaschuk et al., 2007, J Pharm Sci, 96(1)

Ha et al., J Pharm Sci. 2002 Oct;91(10):2252-64.

Improper storage and handling can result in oxidative

degradation of PS. Proteins can be oxidized too, with

potential impairment of binding/efficacy

Trp oxidation as a result of improper storage and handling of PS

(Lam et al., Pharm Res. 2011, 28(10):2543-55)


Importance of raw material testing and storage

Sourcing

Supplier qualification

Batch control

Handling and storage recommendations

Store at 2-8 °C

Protect from air

Protect from light

Single-use containers (avoid re-use after opening)

Kishore et al., 2011, Pharm Res., 28:1194





development

Adherence to GMP


In-process control


Stability testing


What could happen?

Decrease in surfactant content during storage

Loss of protection against interfacial stress

Not necessarily major impact on protein stability – some

degradation products also surface active

Effect of surfactant degradation products on the

protein

Protein oxidation – the presence of oxidative species may

result in oxidation of e.g. Met, Trp

Possible impact on protein stability by e.g. free fatty acids

A lot!


Impact of PS degradation on protein formulations

Degradation products of PS still show surface activity even after 60% loss of content by micelle assay



What could happen?

Particles!

Poorly soluble degradation products (e.g. fatty acids) may form

particles

>3 ppm lauric acid triggered Protein Particle formation and increase

in soluble aggregates






development

Adherence to GMP


In-process control


Stability testing


Process development, validation and in-process control

Surfactants can adsorb to contact surfaces e.g.

manufacturing equipment (filters, tubing, etc.), leading to

significant losses

Importance of process development and IPC testing

Mahler et al., (2010) J. Pharm. Sci.





development

Adherence to GMP


In-process control


Stability testing


Specifications, release and stability testing

“Quality should be built into the product, and testing alone

cannot be relied on to ensure product quality”(FDA Guidance for Industry: Quality Systems Approach to Pharmaceutical CGMP Regulations)

Know your product!

Surfactant testing for release and stability typically done in the

“extended characterization” assay panel provided that:

Raw material qualification and control is performed

Raw material has been properly stored and handled

Careful and sound drug product and manufacturing process

development has been done – mitigate the risks!

Careful and sound drug product and manufacturing process

characterization has been done

In order to support all of the above…


…extensive analytical toolbox and expertise

are needed


Thank you

Acknowledgements

Roman Mathäs, Dorothee Lay, Susanne Jörg, Monika

Geiger, Dorota Roberts, Stephanie Ferrell, Karen Fallen,

Hanns-Christian Mahler

URL www.lonza.com/DrugProduct | Email [email protected]

http://www.lonza.com/DrugProduct

mailto:[email protected]

Documents

Surfactants: Raw Material and Drug Product Control Strategy · Surfactants: Raw Material and Drug Product Control Strategy Atanas Koulov Lonza Drug Product Services NBC | Boston |