Application of Right Size Testing, Regulatory Responses and Future Directions

Barry Cherney – Amgen Inc.

WCBP January 2016

Overview

• QbD and risk evaluations in an integrated control strategy

• Application of right size testing for legacy products and regulatory responses

• Achieving a future state with newer technologies

• Achieving a future state through risk based decision making

2

An effective and efficient testing strategy utilizes QbD concepts

Characterize

attributes

and perform

risk

assessment

Characterize

process.

Measure the

critical

quality

attribute at

the point

where it is

controlled

Eliminate

redundant

testing

A lean and targeted control strategy eliminates “noise” and puts

focus on what is important to control

Develop

method(s) for

critical

quality

attributes

E.g. MS Based method

(Future State)

A1

A3

A5Attribute

DS Process

Step

1 2 DS

CQA 1 X X

Control strategy integrates all aspects of process and product controls

Production Process

Procedural controls(facility, equipment and operational parameters)

Input controls(raw materials and components)

In-process testing(validation, IPCs, process monitoring)

What is the right size and scope of end product testing?

(specifications, comparability, stability)

Control elements are well coordinated and integrated in

an effective control strategies

4

Scientific risk-based evaluations were used to create a streamlined and focused control strategy

• product characterization

• attribute knowledge

• safety and efficacy assessments

5

Risk Overall The potential

adverse

impact of the

PQA

Severity

The probability of

the PQA falling

outside of

acceptable

ranges due to

deviations

Occurrence

The detection

capability and

stringency of the

tests applied.

Detection

, ,

• process

characterization

• challenge studies

• degradation

studies

• historical data

• method sensitivity and

specificity

• testing frequency

(i.e. lot release vs.

comparability

• limits

• downstream

redundancy

General Principles for Revise Existing Specifications

• Removal of tests for impurities with well understood mechanisms for removal and proven process capability

• Removal of redundant tests that are performed at multiple points in the process

• Move testing to the point at which it is controlled• Apply Real-Time-Release-Testing (RTRT) where appropriate

• Removal of tests for quality attributes that are well controlled during manufacturing and where adequate detections are in place to identify issues

• Removal of stability tests for attributes that are not impacted by relevant mechanisms of product degradation

Maintain the ability to perform tests as needed

e.g., for comparability and non-conformance investigations

APPLICATION OF RIGHT SIZE TESTING AND REGULATORY

RESPONSES

7

GENERAL REGULATORY FEEDBACK

• Multiple regulatory applications were submitted to the US FDA (8)

• Limited number of submissions to EU, HC, TGA, CH, and Emerging Markets

• In General Amgen’s approach was well received by regulators (50-100% of proposals accepted)

• US FDA was more restrictive than the other jurisdictions which accepted most if not all of our proposals

• We were more successful eliminating DS testing rather than DP release and stability specifications

• Low probability of an occurrence did not trump the need for a sensitive method of detection

8

Removal of tests for process related impurities with well understood mechanisms for removal and proven process capability for clearance (eg., CHOP, DNA, Protein A)

CHO Protein in VI Pool

CHOP Challenge Study Demonstrates

Excess Clearance Capacity at Column 3

Process Step Normal Process Challenge

Study Results

Column 1 pool 2500 to 3300

ng/mg

2148 ng/mg

VI pool 1 to 2 ng/mg Skip

Column 2 pool 0 to 1 ng/mg Skip

Column 3 pool NT 8.3 ng/mg

(2.4 LRV)

Historical Data

PEGylated Protein:Removal of Redundant Tests for the Same Attributes

• SDS-PAGE gels and SE-HPLC are redundant for detecting the attributes of interest, detection assured through SE-HPLC

10

Regulatory response was mixed, FDA cited multiple cases where unexpected protein

clips occurred during routine manufacture. SEC was not sufficiently robust

Current Optimized

CQ

A1

Method

Release Stability

DS DP DS DP

SEC X X X X

Gel X X X X

CQ

A1

Method

Release Stability

DS DP DS DP

SEC X X X X

Gel

]

Removal of redundant tests and movement of testing

to the point nearest where the attribute is controlled

11

CQA is determined early in DS process; does not change through further

purification, DP processing, or storage

Attribute

Process Step Stability

Col2

Pool

DS DP DS DP

CQA 1 X X X X X

Attribute

Process Step Stability

Col2

Pool

DS DP DS DP

CQA 1 X

Current Optimized

Column 2 Pool vs. DP DP Stability

Removal of redundant tests that are performed at

multiple points in the process and move testing to the

point at which it is controlled

12

CE-HPLC lot purity is determined early in the DS process

AttributeProcess Step

VI Pool DS DP

CQA 1 X X X

AttributeProcess Step

VI Pool DS DP

CQA 1 X

Current

Right Sized

VI Pool vs. DP

Regulatory response was positive for moving the test from DS release to in-process.

Removal from DP testing was questioned due to the assay’s ability to detect other

potential degradation products.

Move DP testing to in-process at the point at which it is last controlled (real time release)

• Protein concentration

• Deliverable volume

• pH

• Osmolality

• Polysorbate

• Appearance, Color, Clarity

• Product related variants

13

Proposed product potency testing removed from DP specifications

14

Overlay Plot of DS and DP Potency Histogram of DS and DP Potency Paired Differences

Summary of Testing Assessment

• DS is tested with no significant change in product potency between DS and DP

under normal processing conditions

• Characterization studies demonstrate that DP process stresses are not expected to

impact potency, critical operating parameters are well controlled

• For all products, changes in potency would be detected by assays that are retained

on the DP specification and monitoring program would initiate investigations if OOT.

For Product X Aggregation is the primary degradation

product with the potential to impact potency. Changes

are more readily detected by SE-HPLC

15

Impact of Force degradation at 60oC of mAb X

Assessed by SE-HPLC and Bioassays

0

20

40

60

80

100

120

0 1 2 3 4 5 6 7 8

Days at 60°C

% R

ela

tive P

ote

ncy

0

20

40

60

80

100

120

% M

ain

Peak

HTRF

Reporter Gene

TRAP

SEC

85

90

95

100

105

110

115

120

0 0.5 1 1.5 2 2.5 3 3.5

% R

ela

tive

Po

ten

cy

Time (Months)

HTRF Bioassay A

98.2

98.4

98.6

98.8

99

99.2

99.4

99.6

99.8

0 0.5 1 1.5 2 2.5 3 3.5

% M

ain

Pe

ak

Time (Months)

SE-HPLC Main Peak B

Impact of Accelerated Thermal Storage Condition at 37C of

mAb X Assessed by HTRF Assays (Panel A) and SE-HPLC

(Panel B)

Regulatory response was mixed, FDA cited regulatory requirements (21 CFR 610.10

and 600.3(s)) and the need to confirm protein conformation for a therapeutic DP

protein. The proposal was accepted by other jurisdictions.

Removal of stability tests

16

Use statistical tools to identify attributes that do not change during storage

% M

ain

Peak

65

70

75

80

85

90

Time (months)

0 6 12 18 24 30 36 42 48

AOFdata.CEXds

___ Predicted Mean __ __ 95% Confidence Bound on Mean

Lot Number

0010023946001003145900100556690010067117001006807600100683330010068338001008852800100893110010110845001011488100101183700010129046001014262400101478440010148022001014850400101557720010173317049C048022049C048023049C080952049D106926049D108163

Lower Spec = 65

Recommended

Storage -30 C

Accelerated

Storage 5 C

No practically important changes over time.

Removal of potentially stability-indicating assays was questioned by FDA even when

no change observed at the recommended or accelerated conditions to monitor for rare

unexpected events

Removal of stability tests for attributes that are not impacted by the known mechanisms of product degradation and do not change over time

Current State:

(Red—assays to remove)

• Appearance, color, clarity

• CE-HPLC

• SE-HPLC

• rCE-SDS/SDS PAGE

• Bioassay

• pH

• Protein Concentration

Optimized:

(Blue- not approved in all regions)

• SE-HPLC

• rCE-SDS

• Bioassay

• pH

• Appearance

18

Stability Testing for Drug Substance

Optimized*:

(Blue- not approved in all regions)

• Appearance (Occasionally retained )

• Purity CE-HPLC; RP-HPLC

• SE-HPLC

• rCE-SDS/SDS PAGE

• pH (Occasionally retained)

*Lyo product optimized

Removal of stability tests for attributes that are not impacted by the known mechanisms of product degradation and do not change over time

Current State:

(Red—assays to remove)

• Appearance, color, clarity

• CE-HPLC

• SE-HPLC

• rCE-SDS

• Bioassay

• pH

• Protein Concentration

Optimized:

(Blue- not approved in all regions)

• SE-HPLC

• rCE-SDS

• Bioassay

• pH

• Appearance

19

Stability Testing for Drug Product

Optimized:

(Blue- not approved in all regions)

• Appearance

• CE-HPLC/ RP-HPLC

• SE-HPLC

• rCE-SDS

• Bioassay

• pH

Introducing Newer Technologies Achieving a Future State

20

Achieving a Future State for Biologics

• Knowledge of attribute criticality is improving through scientific studies and clinical experience

• High resolution analytical capabilities such as MS provide detailed characteristics of critical protein attributes

• Use of attribute specific control levers to control critical product attributes during manufacturing

• Through more direct measurement and control during processing, we will deliver safe and efficacious biopharmaceuticals with well defined attributes

21

Thermo Exactive (Orbitrap MS)

• Thermo Exactive (Orbitrap Family)

• Hi resolution

• Fast scan speed

• Small footprint

• Technology allows for “Plug and Play” peptide map analysis

• Pinpoint Software provides the ability to generate a comprehensive attribute target list and automated quantification.

• Alignment of method and instrumentation for process development, PAT control and product disposition

. 22

Replacing non-attribute specific assays with methods capable of specifically detecting and measuring critical attributes.

CEX separation

70% Potency

Attribute 1

Attribute 2

Attribute 3

Attribute 4

Current

Release Method

150% Potency

Attribute 5

100% Potency

Main peak

Product

UnderstandingFuture

Release Method

Sub-fraction & BRQA

Assessment

CEX separation

MS Based method

A1

A3

A5A1, A2

2 x A3

A3, A4

A5

Attribute Potency

*

Safety

Main

peak

100% 1

A1 50% 3

A2 110% 3

A3 95% 7

A4 102% 1

A5 150% 5

Regulatory Argument:

Replacing CEX monitoring

of pre-peaks with more

specific method monitoring

relevant attributes:

• A1 (efficacy)

• A3 (safety)

• A5 (safety and efficacy)

Single Multi-Attribute method is able to directly monitor more attributes than all other conventional methods combined

24

Multi-Attribute Method Conventional Release Methods

Antibody PQA Pep Map-MS SEC CEX rCE-SDS nrCE-SDS HILIC ID ELISA HCP ELISA

Aggregate Assessment N Y Indirect Y Y N N N

Deamidation (Isomerization) Assessment Y N Indirect N N N N N

Disulfide Isoform Assessment maybe N Indirect N Y N N N

Glycation Assessment Y N N Y Y N N N

High Mannose Assessment Y N N N N Y N N

Methionine Oxidation Assessment Y N N N N N N N

Signal Peptide Assessment Y N N N N N N N

Unusual Glycosylation Assessment Y N Indirect maybe maybe Y N N

CDR Tryptophan Degradation Assessment Y Indirect N N N N N N

Non-consensus Glycosylation Assessment Y N N maybe maybe N N N

N-terminal pyroGlutamate Assessment Y N Indirect N N N N N

C-terminal Lysine Assessment Y N Y N N N N N

Galactosylation Assessment Y N N N N Y N N

Dimer Assessment N Y N N N N N N

Fragmentation (peptide bond) Assessment Y maybe N Y Y N N N

Disulfide Reduction (DS Fragmentation) Assessment maybe N N N Y N N N

Host Cell Protein Assessment Y N N N N N N Y

Mutations/Misincorporations Assessment Y N N N N N N N

Hydroxylysine Assessment Y N N N N N N N

Thioether Assessment Y N N N N N N N

Trisulfide Assessment maybe N N N N N N N

Non-glycosylated Heavy Chain Y N N N N N N N

DNA Assessment no N N N N N N N

Cysteine Adducts Assessment maybe N maybe N N N N N

C-terminal Amidation Assessment Y N Indirect N N N N N

CDR Conformers (HIC Isoform) Assessment no N Indirect N N N N N

O-linked glycans Assessment maybe N N N N N N N

Fucosylation Assessment Y N N N N N N N

Residual Protein A Y N N N N N N N

Identity Y N Y N N N Y N

which PQA’s to monitor and control based on risk to PQ

Multi-Attribute Method (MAM)

• Provides direct monitoring of PQA’s compared to indirect monitoring by conventional methods (ie CEX)

• Allows one to focus on critical PQA’s thus assessing the risk associated with biologically relevant attributes

• Eliminates conventional release assays

• CEX

• CE-SDS

• ID test

• HPLC

• HCP ELISA

• PrA ELISA

Attribute Focused Acceptance Criteria

*Average purity for historical lots

Many product specifications report %main peak - ambiguous with regards to the

CQA’s being controlled. Proposal to focus on the CQAs

Attribute Current State Parameter Future State Parameter

Size

Variants

SEC %Main peak Not reported

SEC % HMW SEC %HMW (dimer* and

aggregate*)

rCE-SDS %HC +%LC rCE-SDS %LMW+%MMW

Charge

Variants

by CEX or

cIEF**

%Main peak Not reported

% Acidic peaks: ‘report’ or

‘comparable to RS’

% Acidic peaks*: specify range

% Basic peaks: ‘report’ or

‘comparable to RS’

% Basic peaks*: specify range

Achieving a “future state: through risk based decision making and reliance on

the PQS

27

Even in the era of QbD, non-Risked based rejections are still common

• Setting limits well below a possible adverse impact to PQ as it relates to S & E (eg., endotoxin)

• Setting limits for variants that are different for each SKU (eg., metox AC varies from 5% to 17%)

• Setting limits for new peaks without any understanding of the risk

• Setting limits based on non specific information

• SVPs by HIAC (no info on nature of the SVP)

• HCP by Elisa (no info on nature of the protein

• CE-HPLC (multiple variants with various level of risk

Investigational limits to determine the actual risk to PQ should be established when additional information can inform a decision.

28

Embracing a science and risked based control strategy

• QbD is making an impact but specification setting has lagged behind – mostly were still setting limits based on process capability with input from clinical experience

• Monitoring process capability is an important tool for continuous process understanding and improvement but should not be the basis for a product rejection

• Industry and regulators have to fully embrace risked based control strategies and move away from reliance on limited clinical experience and/or process consistency

29

Conclusions

• An integrated, risk-based control strategy utilizing PQA understanding and process knowledge offers the opportunity for right size testing providing control of critical attributes at the appropriate point in the process

• High resolution analytics will make many current methods less informative and redundant

• Implementation of QbD (including Q10) permits increased reliance on PQS to investigate and make appropriate decisions for patients rather than reject product that is suitable for its intended use

30

Acknowledgements

• Jay Higgins

• Bob Bailey

• Tony Mired-Sluis

• Karen Miller

• Chantal Cazeault/ Darrin Cowley

• Julie King/ John Rolf/ Simon Szeto /Barbara Rellahan

• Roger Hart

• Tom Monica

• Wendy Laderach

31