We will introduce an excipient platform that makes it possible to combine excipients in ways that can reduce protein viscosity to a greater extent.

1. The life science business of Merck KGaA,
Darmstadt, Germany operates as
MilliporeSigma in the U.S. and Canada.
The Viscosity Reduction Platform:
Enabling Subcutaneous Delivery
Darmstadt, March 22nd 2022
Dr. Tobias Rosenkranz

2. The life science business
of Merck KGaA, Darmstadt,
Germany operates as
MilliporeSigma in the U.S.
and Canada

3. Challenges arising from
elevated protein viscosity
What we offer
Our solution: The Viscosity
Reduction Platform

4. Challenges arising from
protein viscosity

5. Increasing protein concentrations can lead to high viscosity levels
beyond injectability
Viscosity Reduction Platform
Market Need
 Most therapeutic mAbs are currently only
available for intravenous (IV) administration
(64%)
 Subcutaneous (SC) formulations are the
preferred route, as they offer higher patient
convenience and the opportunity to reduce
healthcare cost
Challenge
 SC formulations can only reach volumes in the
range of 1-2 mL
➢Need for high protein concentrations to reach
therapeutic dose
 High protein concentrations tend to exhibit
viscosity levels beyond injectability
Webinar - The Viscosity Reduction Platform: Enabling Subcutaneous Delivery | March 22, 2022
5

6. Effect of viscosity in highly concentrated antibody formulations
Viscosity Reduction Platform
Numbers
The required injection force depends on various
parameters other than viscosity
η: viscosity [mPa*s]
l: length of needle [mm]
Q: flow rate/injection rate [mL/s]
Rb: radius of syringe/barrel [mm]
Rn: inner radius of needle [mm]
Typically, needles thinner than 27G are used for sc
administration.*
*Garindel & Presser, Lyophilization of High Concentrated Protein Solution, Book
Chapter: Lyophylization of Pharmaceuticals and Biologics, Springer Protocols 2019
6
Injection force is
dependent on
solution viscosity
Webinar - The Viscosity Reduction Platform: Enabling Subcutaneous Delivery | March 22, 2022

7. 7 Webinar - The Viscosity Reduction Platform: Enabling Subcutaneous Delivery | March 22, 2022
Viscosity Reduction Platform
Effect of protein viscosity on injection force for mAb vs. water
27 Gauge Needle
Water: 1 N corresponds roughly
to the force of 100 g
27 Gauge Needle
mAb: 80 N corresponds roughly
to the force of 8 kg
A thinner needle would additionally amplify the required
injection force !

8. Viscosity Reduction Platform
Several mAbs show a critical viscosity vs. concentration profile
In-house mAbs and marketed mAbs identified as models with viscosity issues
Webinar - The Viscosity Reduction Platform: Enabling Subcutaneous Delivery | March 22, 2022
8

9. Viscosity Reduction Platform
Molecular mechanisms causing viscosity in proteins: Fab-Fab interactions
Antibody regions Viscosity at protein concentrations < 200 mg/ml
 Caused by protein-protein interactions (PPI)
 PPI can occur between different regions of the protein,
however Fab-Fab interactions seem predominant*
 Interactions are specific, but transient
 PPI are identical to intra-molecular interactions that stabilize
proteins
*S. Kanai et al. Journal of Pharmaceutical Sciences, Vol. 97, No. 10, October 2008
Antibodies comprise of different regions
Fab: Fragment antigen binding
Fc: Fragment crystallizable
9 Webinar - The Viscosity Reduction Platform: Enabling Subcutaneous Delivery | March 22, 2022
Fab
Fc

10. Webinar - The Viscosity Reduction Platform: Enabling Subcutaneous Delivery | March 22, 2022
Single excipients often not sufficiently able to reduce protein viscosity
Viscosity Reduction Platform
A single excipient
may not reduce
viscosity
sufficiently well,
even when increasing
the excipient
concentration
10
BM: Benchmark BSAcid: Benzene sulfonic acid
Orn: L-Ornithine HCl Pyr: Pyridoxin
Phe: L-Phenylalanine TMP: Thiamine phosphoric acid ester

11. Webinar - The Viscosity Reduction Platform: Enabling Subcutaneous Delivery | March 22, 2022
High single excipient concentrations can destabilize the protein
Viscosity Reduction Platform
Forced degradation study
• 120 mg/ml Infliximab; 170 mg/ml Evolocumab
• 40°C, 75% rel. hum., 28 days
• Reference formulation vs. formulations with
excipients spiked in
Higher excipient concentrations can
destabilize the antibody
11

12. Our solution: Viscosity
reducing Excipients

13. Use of excipient combinations to reduce protein viscosity efficiently
Viscosity Reduction Platform
Our viscosity reducing excipient platform:
A set of excipients to help you manage protein viscosity
• Numerous excipients for reducing viscosity are already
known and used in marketing formulations, e.g. certain
amino acids
• Known excipients are not always able to reduce protein
viscosity sufficiently well
• Used in combination, excipients may develop synergistic
effects enabling improved viscosity reduction and a better
balance of viscosity vs. stability
Webinar - The Viscosity Reduction Platform: Enabling Subcutaneous Delivery | March 22, 2022
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14. Webinar - The Viscosity Reduction Platform: Enabling Subcutaneous Delivery | March 22, 2022
Viscosity Reduction Platform
Effect of protein viscosity on required injection force for mAb
Infliximab
27 Gauge Needle, 0.2 ml/s flow rate
Aspiration Time: 75 s
Injection Force: 19 N
Evolocumab
27 Gauge Needle, 0.2 ml/s flow rate
Aspiration Time: 116 s
Injection Force: 29 N
Typical flow rates 0.15 ml/s – 0.45 ml/s
Usach, I et al.; Subcutaneous Injection of Drugs: Literature Review of Factors
Influencing Pain Sensation at the Injection Site; Adv Ther (2019) 36:2986–2996
14
Aspiration time Injection Force Aspiration time Injection Force

15. Combinations of excipients enable SC delivery
Viscosity Reduction Platform
Webinar - The Viscosity Reduction Platform: Enabling Subcutaneous Delivery | March 22, 2022
Several useful combinations depending on the properties of the
protein under the formulation conditions
15

16. Webinar - The Viscosity Reduction Platform: Enabling Subcutaneous Delivery | March 22, 2022
Viscosity Reduction Platform
Effect of protein viscosity on required injection force for mAb
Infliximab
27 Gauge Needle, 0.2 ml/s flow rate
Aspiration Time: 42 s - 44%
Injection Force: 15 N - 21%
Typical flow rates 0.15 ml/s – 0.45 ml/s
Usach, I et al.; Subcutaneous Injection of Drugs: Literature Review of Factors
Influencing Pain Sensation at the Injection Site; Adv Ther (2019) 36:2986–2996
16
Aspiration time Injection Force

17. Webinar - The Viscosity Reduction Platform: Enabling Subcutaneous Delivery | March 22, 2022
Viscosity Reduction Platform
Effect of protein viscosity on required injection force for mAb
Infliximab
27 Gauge Needle, 0.2 ml/s flow rate
Aspiration Time: 42 s - 44%
Injection Force: 15 N - 21%
Evolocumab
27 Gauge Needle, 0.2 ml/s flow rate
Aspiration Time: 37 s - 68%
Injection Force: 15 N - 48%
17
Aspiration time Injection Force
Aspiration time Injection Force
Typical flow rates 0.15 ml/s – 0.45 ml/s
Usach, I et al.; Subcutaneous Injection of Drugs: Literature Review of Factors
Influencing Pain Sensation at the Injection Site; Adv Ther (2019) 36:2986–2996

18. Webinar - The Viscosity Reduction Platform: Enabling Subcutaneous Delivery | March 22, 2022
Viscosity Reduction Platform
Excipient combinations show an improved stability
Use of excipient combinations can counteract the destabilizing effect of
strong viscosity reducing excipients !
18
Forced degradation study
• 120 mg/ml Infliximab; 170 mg/ml Evolocumab
• 40°C, 75% rel. hum., 28 days
• Reference formulation vs. formulations with excipients spiked in

19. Webinar - The Viscosity Reduction Platform: Enabling Subcutaneous Delivery | March 22, 2022
Viscosity Reduction Platform
Excipient combinations show an improved stability
Storage and accelerated storage study:
• At 2-8°C high degree of stability for the relevant excipient combinations
19

20. Webinar - The Viscosity Reduction Platform: Enabling Subcutaneous Delivery | March 22, 2022
Viscosity Reduction Platform
Excipient combinations show an improved stability
Storage and accelerated storage study:
• At 2-8°C high degree of stability for the relevant excipient combinations
• At 25°C degradation of protein with TMP observed
• Likely reason is light/temperature sensitivity of excipient
20

21. Benefit of our platform of synergistic excipient combinations
Viscosity Reduction Platform
Viscosity reduction
Improved administration
• Reduced injection volume / less frequent injections
• Switch to more convenient route of administration (SC)
Increased patient
convenience and compliance
Webinar - The Viscosity Reduction Platform: Enabling Subcutaneous Delivery | March 22, 2022
21
Protein stability

22. Viscosity Reduction Platform
Webinar - The Viscosity Reduction Platform: Enabling Subcutaneous Delivery | March 22, 2022
Enable
subcutaneous
delivery
Increase
dose per volume
ratio
Improve
process
economics
22

23. What we offer

24. Webinar - The Viscosity Reduction Platform: Enabling Subcutaneous Delivery | March 22, 2022
How to evaluate our
technology
• Test-Kit available on request
(technical grade)
• Detailed user guide describing
proof-of-concept screening
available
24

25. Formal Paper
Toxicological Profile
and
Safety Evaluation
by European
Registered Toxicologist
In-vitro
skin irritancy test
(HET-CAM)
No irritancy
detected
Available on request
25
Regulatory and toxicological considerations positive
Viscosity Reduction Platform
Well characterized safety profile
Toxicological evaluation Prior in-human usage
Excipient candidates have all
been used in parenteral
formulations:
• Parenteral nutrition
(here considered
actives)
• Counter ions to APIs
• pH-adjustment
Webinar - The Viscosity Reduction Platform: Enabling Subcutaneous Delivery | March 22, 2022

26. 26 Webinar - The Viscosity Reduction Platform: Enabling Subcutaneous Delivery | March 22, 2022
Summary
Enabling subcutaneous delivery
• Excipient platform can specifically target viscosity challenges
• Use of excipient combinations is more efficient than using single
excipients
• Aspiration time and extraction force can be significantly reduced
• Use of excipient combinations offers additional stability benefits
• Dose concentration can be increased
• A reduced protein viscosity can yield improved process efficiency
For more information, please visit:
sigmaaldrich.com/viscosity-reduction

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