excipients: critical components of the biomanufacturing ...€¦ · meets shelf-life objectives ......

Jeffrey L. Shumway

MilliporeSigma

Excipientfest – Providence, RI

April 26, 2017

ExcipientS: Critical Components of the BiomanufacturingProcess

Agenda

The Bioprocess Perspective1

Administration & Drivers of Biomolecule Therapeutics2

Functionality of Excipients: Stability3

Aggregation & Contributing Factors4

Sources of Impurities & Risk Mitigation5

Additional Process & Excipient Considerations6

Practical Considerations7

Summary & Discussion8

CLASSIC PERSPECTIVE

4

The Bioprocess Perspective

FORMULATION

PURIFICATION

A DIFFERENT PERSPECTIVE

CONTINUOUS FORMULATION

5

Chemistries of the recombinant/mAb process and formulation

Protein refolding

Virus Inactivation Chromatography Formulation

Urea

Ultra/Diafiltration

Glutathione

Arginine/Cysteine

Ammonium sulfate

Acetate

Citrate

Glycine

Triton™ X-100

Tri-N-Butylphosphate

Tris/Tris HCl

Glycine

Ammonium sulfate

Sodium Chloride

Urea

Ethanol (storage)

Sorbitol/Mannitol

Sucrose

Sodium Chloride

Histidine

Polysorbate 20/80

Citrate

Acetate Acetate

Citrate

Phosphate Phosphate

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7

Administration of biomolecule therapeutics

Parenteral Administration• Intravenous• Direct Injection

• Infusion

• Subcutaneous

• Intramuscular

• Intradermal

• Intraspinal

• Intrathecal

• Intra-arterial

• Others

8

Administration advantages and challenges of biomolecules

Administration Form Advantages Challenges

Intravenous (IV) • Suitable for substances that may cause irritation

• Large volume administration

• Trained personnel; qualified environment• Port system requirement• Location of peripheral cannula• Time-consuming administration• Risk of systemic infection

Subcutaneous (SQ) • Short clinical visit• Lower resource burden• Less invasive (as compared to IV)• Self-administration possible

• Pain-free administration of larger volumes• Minimization of effects at injection site• Adsorption and bioavailability• Administration of exact dose

Geburtshilfe Frauenheilkd. 2014 Apr; 74(4): 343–349.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4078128/

High Concentration Drivers

High patient doses required for biological products: ~0.1-3mg/kG body weight

Intravenous (IV) infusion most common delivery method due to high administered

doses and low protein concentration

Subcutaneous administration using prefilled syringes preferred by patients

Ease of Use

Fast

Low cost

Dani, 2007

10

Formulation Criteria

Simplicity

Manufacturing-friendly

Robust

Safe (use appropriate excipients)

Meets shelf-life objectives

Satisfy target profile

Meets time-lines

“Informed Compromise”

Rational/Defensible

Reference: “Introduction to the Stabilisation and Formulation of Proteins and Peptides”; Legacy BioDesign, LLC

Agenda









Stability

Conformational

•Native vs. Denatured states

•Chemical Denaturation

•Thermal Denaturation

Colloidal

•Soluble vs. Insoluble proteins

•Distinctly different from aggregation

•Native-state association

Interfacial

•Protein Adsorption

•Binding results in physical instability

•May include interaction with air-water interface

12

Conformational Stability

Stabilizers in solution (i.e. polyols, sugars, etc)

Sugars are also known to stabilize protein conformations against dehydration stresses by substituting surrounding water molecules through hydrogen bonding

Excluded solvent behaviors

13

Sugar/Polyol Initial Concentration Typical Range

Trehalose 0.5 M 0-1.0 M

Mannitol 2% w/v

Sorbitol 0.5 M 0.2-1.0 M

Sucrose 0.5 M 0.2-1.0 M

Amino Acids

Glycine 250 mM 0.25-2.0 M

L-Histidine HCl 20 mM

L-Histidine 20 mM

Lebendiker & Danieli, FEBS Letters 588 (2014) 236–246

Colloidal Stability

Proteins exhibit colloidal properties in solution Attractive or repulsive interactions

Interactions effect solution properties

Solubility, viscosity, and crystallization – can contribute to aggregation

14

Tonicity Agents/Mineral Salts Initial Concentration Typical Range

Sodium Chloride 300 mM 0-1 M

Sodium Sulfate 500 mM 0-0.5 M

Potassium Chloride 200 mM 0-1 M

Ammonium Sulfate 50 mM 0-0.2 M

Amino Acids

L-Arginine HCl 125 mM 0-2 M

L-Arginine 125 mM 0-2 M


Interfacial Stability

Surfactants

General interaction: Lower the surface tension between two media (liquid/liquid, liquid/solid, liquid/gas)

Non-ionic Detergents

Characterized by non-ionic, hydrophilic headgroups

Limits interaction with ions, hydrophobic environments in solution

15

Surfactant Initial Concentration Typical Range

Polysorbate 20 0.1% 0.01-0.5%

Polysorbate 80 0.1% 0.01-0.5%


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17

What are Aggregates?

1. Mahler, H.C.; Jiskoot, W.; “Analysis of Aggregates and Particles in Protein Pharmaceuticals. Hoboken, NJ: John Wiley & Sons, 2012.

• Aggregates are multiple drug molecules held together by covalent bonds or intermolecular forces

• Aggregates larger than pentamers and large particles are of particular interest

• Measurement via multiple methods1

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Therapeutics with high aggregate concentration administered by IV or subcutaneous injection can cause an immune reaction

- Safety Concern – Critical Quality Attribute

- Stresses weakened patient

- Can lead to reduced therapeutic efficacy

Regulatory requirements:

- Testing for visible and subvisible particles

- Maximum specifications

Why are aggregates important?

Rosenberg, A. S. “Effects of Protein Aggregates: An Immunologic Perspective” AAPS Journal 2006, 8 (3).

Carpenter, J. F.; Randolph, T. W.; Jiskoot, W.; Crommelin, D. J.; Middaugh, C. R.; Winter, G.; Fan, Y.-X.; Kirshner, S.; Verthelyi, D.; Kozlowski, S.; Clouse, K. A.; Swann, P. G.; Rosenberg, A.; Cherney, B. “Overlooking Subvisible Particles in Therapeutic Protein Products: Gaps that may Compromise Product Quality” Journal of Pharmaceutical Sciences2009, 98 (4), 1201–1205.

19

Factors Influencing Aggregation

Stresses on protein:

• pH and salt

• Mechanical/Shear

• Raw material quality

Can be improved by selecting the

appropriate excipients, of the appropriate

quality

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Drug Substance Drug Product

Formulation: Drug Substance + Excipients = Drug Product

Formulation

www.fda.gov

Source of Impurities:

Bulk Drug Substance purification process

Risk-Mitigation Strategies

22

Quality of Raw Materials

• Quality of raw materials can influence protein stability and aggregation

• Stringent quality control can reduce raw material-associated risks

• All specification parameters tested on every raw material batch

• Test methods are thoroughly validated

• Raw material storage stability tested

Impurity Profile• Elemental Impurity Profile is not

required of excipients

• Elemental Impurity Profile is requiredof Drug Product

• ICH Q3D: The applicants risk assessment can be facilitated with information about potential elemental impurities (e.g. by supplier of drug substance and excipients)

• Example shows all 24 elements of ICH Q3D

• Levels above option one limit (calculated to a daily intake of ten gram of drug)

Example: ICH Q3D Elemental Impurity information

23

Example

PDA Formalized Risk Assessment

Excipient Support Documents

• Product quality report [4.1 *]

• Elemental impurity information

• Analytical procedure

In line with CTD chapter 3 quality (adapted for excipients) [2.3*]

• General information

• Manufacture

• Characterization

• Control of drug substance

• Reference standard

• Materials

• Container closure system

• Stability

• Quality Self Assessment [2.6*]

• Audit report summary [2.6*]

• Supply chain Information [2.3*]

• Stability data [2.3*]

[*] No. of supported chapter of Guideline on formalized risk assessment for excipients EU/C 95/210 24

Material qualification

Risk assessmentProcess optimization

Minumum information required to begin material assessment

Support information during Formalized Risk Assessment

Support for Process Optimization and evolving

Regulatory landscape

Formalized Risk Assessment

EU 2015/C 95/02

“… the holder of the manufacturing authorization shall ensure that the excipients

are suitable for use ... by verifying the appropriate GMP…”

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Ultrafiltration & Diafiltration Considerations

Ultrafiltration• IS: Increase in macromolecule concentration

• DOES: Typical concentration increases 5-50x

• MEANS: Anything ≥ MWCO will also increase in concentration

Diafiltration• IS: Exchange of solvent/buffer/excipient conditions

• DOES: Requires 7-10 diavolumes in practice

• MEANS: Anything ≥ MWCO will also increase in concentration

Contaminants such as pyrogens (MW 10kDa-100kDa) can increase in concentration as much as 12-60x in worst case scenario

Retentate

PermeateFeed

Diafiltrate

10/30kDa MWCO

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Mechanical/Shear Stress

Mechanical/Shear stress can result in largely increased liquid-air interface (bubbles, foam)

Exposure to the liquid-air interface is an aggregation risk for the therapeutic protein!

Stabilizers can protect therapeutic proteins from such stresses in process and final formulated product

Stabilizer options:

Sugars, Polyols and Amino Acids• Conformational stabilizers: Reduce sensitivity to

stress at interfaces by forcing the protein into tight

conformation

Surfactants• Interfacial stabilizers: Reduce stress exposure

of the protein at liquid-air/solid interface

• Almost exclusively applied in the final formulation

due to process challenges

Stirring PumpingShaking

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Practical Considerations for Bioprocessing Excipients

Process-friendly

Specialized requirements

• Microbial qualification

• Endotoxin qualifcation

• Low levels of contaminants

• Example: Tris, low in polyoxymethylene (POM)

• GMP qualified production

• Example: sodium caprylate

• Paper-free packaging

• Storage stability

• Clumping/flocculant excipients

• Custom packaging

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Agenda









31

Biologics require special consideration

Biopharmaceuticals• Trend toward higher concentration biologic

therapeutics

• Formulation excipients can prevent protein aggregation, both in the drug product and process

• Excipient grade formulation additives are higher value – rational design of formulations is a must

• Effective chemistries prevent product-product interactions or prevents shear damage

• Trace contaminants such as heavy metals can effect protein stability (elemental impurities)

CLASSIC PERSPECTIVE

32

The Bioprocess Perspective

FORMULATION

PURIFICATION

A DIFFERENT PERSPECTIVE

CONTINUOUS FORMULATION