Critical Aspects of Synthetic Lipids Drug Development

Merck KGaA
Darmstadt, Germany
Judit Huarte Ciganda, PhD
Technical Product Manager Synthetic Lipids
Critical aspects and strategies

The life science business of
Merck KGaA, Darmstadt, Germany
operates as MilliporeSigma
in the U.S. and Canada.
Successful drug development with synthetic lipids: Critical aspects and strategies | May 24, 20182

01 Lipids: role in liposomal formulation
02 Synthetic lipids: process development
03 Regulatory aspects
04 Troubleshooting: examples
05 Enhanced surface characteristics
06 Overview of facilities and capabilities
3

4

Introduction
Lipids: What makes them special?
5 Successful drug development with synthetic lipids: Critical aspects and strategies | May 24, 2018
hydrophilic head hydrophobic tail
1,3-GDO
(R)-DODMA
DPPC

Liposome rigidity
Phase transition temperature
Liposome leakage
Liposome shape
Introduction
Lipid-based structures in an aqueous environment
BILAYERED MONOLAYERED
LIQUID CORE
SOLID CORE

Surface charge, polarity
Liposome shape
Liposome size
Cell affinity
Introduction
Lipid-based structures in an aqueous environment
Non-targeted Targeted
Non-PEGylated
PEGylated

Introduction
Liposome manufacture
Lipids in organic solvent
(and hydrophobic drug)
Drying step Hydration
Addition of water
(and hydrophilic drug)
Stirring
Sonication
Extusion
Homogenization
Multilamellar vesicles Unilamellar vesicles
Lipid film / cake
+ Purification
Mechanical methods
• Shaking of phospholipid dispersions
• Extrusion through a filter (low/medium
pressure)
• Extrusion through a French press cell
(Microfluidizer technique)
• High-pressure homogenization
• Ultrasonic irritation
• Bubbling of gas
Methods based on organic
solvent replacement by
aqueous media
• Organic solvent removal
• Use of water-inmiscible solvents
• Ethanol injection method
• Ether infusion (solvent vaporization)
• Reverse-phase evaporation
Methods based on detergent
removal
• Gel exclusion chromatography
• Slow dialysis
• Fast dilution
• Other related techniques
Wagner and Vorauer-Uhl. „Liposome technology for industrial Purposes“ Journal of Drug Delivery 2011(2010).
Challenge: scalable + robust + efficient processes

Liposomal formulations in the market
9
Product
(Approval Year)
Admin.
route
Active Agent Indication
Doxil® (1995) i.v. Doxorubicin Ovarian, breast cancer, Kaposi’s sarcoma
DaunoXome®
(1996)
i.v. Daunorubicin AIDS-related Kaposi’s sarcoma
Depocyt® (1999) Spinal Cytarabine/Ara-C Neoplastic meningitis
Myocet® (2000) i.v. Doxorubicin Combination therapy with cyclophosphamide in metastatic breast
cance
Mepact® (2004) i.v. Mifamurtide High-grade, resectable, non-metastatic osteosarcoma
Marqibo® (2012) i.v. Vincristine Acute lymphoblastic leukaemia
Onivyde™ (2015) i.v. Irinotecan Combination therapy with fluorouracil and leucovorin in metastatic
adenocarcinoma of the pancreas
Abelcet® (1995) i.v. Amphotericin B Invasive severe fungal infections
Ambisome® (1997) i.v. Amphotericin B Presumed fungal infections
Amphotec® (1996) i.v. Amphotericin B Severe fungal infections
Visudyne® (2000) i.v. Verteporphin Choroidal neovascularisation
DepoDur™ (2004) Epidural Morphine sulfate Pain management
Exparel® (2011) i.v. Bupivacaine Pain management
Epaxal® (1993) i.v. Inactivated hepatitis A virus
(strain RGSB)
Hepatitis A
Inflexal® V (1997) i.v. Inactivated hemaglutinine of
Influenza virus strains A and B
Influenza
Adapted from Bulbake and col. “Liposomal Formulations in Clinical Use: An Updated Review” Pharmaceutics 2017, 9, 12.

Liposomal formulations in clinical trials (1)
10
Product Admin.
route
Arikace Aerosol Amikacin Lung infections
Stimuvax s.c. Tecemotide Non-small cell lung cancer
T4N5 liposomal
lotion
Topical T4 endonuclease V Xeroderma pigmentosum
Liprostin i.v. Prostaglandin E-1 (PGE-1) Restenosis after angioplasty
ThermoDox i.v. Doxorubicin Hepatocellular carcinoma and also recurring chest wall breast cancer
Lipoplatin i.v. Cisplatin Non-small cell lung cancer
Aroplatin i.v. Platinum analogue Metastatic colorectal cancer
Liposomal
annamycin
i.v. Semi-synthetic doxorubicin
analogue annamycin
Relapsed or refractory acute
myeloid leukaemia
SPI-077 i.v. Cisplatin Lung, head and neck cancer
OSI-211 i.v. Lurtotecan Ovarian, head and neck cancer
S-CKD602 i.v. Potent topoisomerase I inhibitor Cancer
LE-SN38 i.v. Irinotecan’s active metabolite Advanced colorectal cancer
LEP-ETU i.v. Paclitaxel Cancer
Endotag-I i.v. Paclitaxel Breast and pancreatic cancers
Atragen i.v. All-trans retinoic acid Hormone-resistant prostate cancer, renal cell carcinoma and acute
myelogenous leukaemia

Liposomal formulations in clinical trials (2)
11
Product Admin.
route
LEM-ETU i.v. Mitoxantrone Various cancers
Liposomal
Grb-2
i.v. Antisense oligodeoxynucleotide growth factor receptor
boundprotein 2 (Grb-2)
Hematologic malignancies
INX-0125 i.v. Vinorelbine tartrate Advanced solid tumours
INX-0076 i.v. Topotecan Advanced solid tumours
TKM-080301 Hepatic intra-
arterial Admin.
PLK1 siRNA Neuroendocrine tumours
Atu027 i.v. PKN3 siRNA Pancreatic cancer
2B3-101 i.v. Doxorubicin Solid tumours
MTL-CEBPA i.v. CEBPA siRNA Liver cancer
ATI-1123 i.v. Docetaxel Solid tumours
LiPlaCis i.v. Cisplatin Advanced solid tumours
MCC-465 i.v. Doxorubicin Metastatic stomach cancer
SGT-53 i.v. p53 gene Various solid tumours
Alocrest i.v. Vinorelbine Breast and lung cancers

Agenda
12

NDA/BLASUBMITTED
Lipid development
Pharmaceutical product development
Graph adapted from Pharmaceutical Research and Manufacturers of America, Biopharmaceuticals
in Perspective, Facts and Figures 2012, (Washington, DC: PhRMA, March 2012).

Synthetic lipids for drug development
What is needed from them?
Convenient and reproducible behavior in formulation: particle characteristics, stability and release profile
High purity will influence:
• lipid stability
• lipid bilayer structure in formulation
• formulation stability
• formulation release profile
1

Lipids purity
How can it be optimized?
LIPID PURITY

Lipids purity
LIPID PURITY
Quality of
starting
materials
High and consistent quality raw materials
• Low level of by-products
• Defined stereochemistry (cis/trans)
• Low bioburden and endotoxin levels
• Plant-derived raw materials with
BSE/TSE and non-GMO certificates
• Use class II and III solvents

Lipids purity
LIPID PURITY
Manufacturing
process
Quality of
starting
materials
The final GMP process needs to be scalable and
reproducible regarding yield and product quality
• Avoid reaction conditions which could lead
to isomerization
• Consider scalability from the very beginning
• Economy of scale, batch size definition
• Reduce number of steps, define GMP steps

Lipids purity
LIPID PURITY
Purification
process
Purification steps must be scalable as well
• Use crystallization if possible
• Employ liquid/liquid extraction methods
• Avoid chromatography
• Convert chromatography into
a filtration over silica gel
Manufacturing
process
Quality of
starting
materials

Consistent quality: In every step of the development
• Avoiding variability in the formulation development process
• Avoiding bridging toxicity studies
Saving resources
1
2
High purity.

Consistent quality.
1
2
3
Good material characteristics: solubility, cristallinity, stability, flowability
 influence on drug product GMP manufacturing process
High purity.

by courtesy of Büchi Labortechnik AG, Flawil, Switzerland
Lipids characteristics
How can they be optimized for an easier formulation process?
The material’s surface can be enhanced by:
• Crystallization
• Lyophilization
• Spray drying
• Milling
• Solubility improvements
• Higher purity
• Enhanced stability
• Easier handling
characteristics
Enabling an easier
formulation
process
GMP relevantAmorphous vs. cristalline material

Lipid development
Process development behind lipid-based formulations
The right choice of raw material will reduce the risk during product development
PROCESS
RESEARCH
• Feasability
• Finding a cost-
effective route
• Initial amounts
manufacture
From months up
to 1 year
PROCESS
OPTIMIZATION
• Economics of process,
yield
• Adjust process to major
safety aspects
• Reliable analytical
methods
• Determination of critical
raw materials
• Initial product stability
studies
1-2 Years
PROCESS
MATURATION
• Process Validation
• Fix raw material
suppliers
• Quality risk analysis
• Final development
reports
• Stability studies for
intermediates
1-2 Years

Regulatory aspects
Successful approval of a lipid-based drug product: which guidelines
should be taken into account?
“[…] The quality of lipid components, including
modified lipids (e.g., polyethylene glycol (PEG)
modified lipids), can affect the quality and
performance of the liposome drug product.
In cases of novel lipid components, the level of
detail provided in the submission should be
comparable to that for a drug substance.[…] see
ICH Q11 Development and Manufacture of Drug
Substances […]”
“[…] The quality and purity of the lipid
starting materials is essential for the later
quality of the drug product, therefore the
appropriate characterization and specification
of the lipid starting material is considered as
vital. […]
The level of information to be provided
with the relevant submission depends on
complexity of the excipients. […]”
“[…] Because the quality of liposome components
such as lipids can affect the quality of whole
liposome drug products, the quality of liposome
components should be appropriately controlled […]
In liposome drug products, lipid components […] and
molecules for liposome modification […] contribute to
an improvement in the in vivo stability,
pharmacokinetics, and intracellular behavior of the
active substance. Therefore, liposome components,
especially ligands (targeting moiety) and antibodies
that have a significant impact on the function of
the drug product, should be evaluated and
controlled to a greater extent than general
excipients to ensure their intended
properties.[…]”

Regulatory aspects
Successful approval of a lipid-based drug product: which guidelines
should be taken into account?
FDA (April 2018)
Liposome Drug
Products: Chemistry,
Manufacturing, and
Controls; Human
Pharmacokinetics and
Bioavailability; and
Labeling
Documentation.
Guidance for Industry.
EMA (2013)
Reflection paper on the
data requirements for
intravenous liposomal
products developed with
reference to an innovator
liposomal product.
NIHS (2016)
Guideline for the
Development of
Liposome Drug
Products.
Site
implementation
ICH Q7 (API GMP) and other ICH Guidelines for Drug
Product and Drug Substance

Regulatory aspects, our approach.
Quality-related deliverables for synthetic lipids

Agenda
27

Troubleshooting throughout the years
Are all those painpoints such a game changer?
Convenient and reproducible behavior in formulation: particle characteristics, stability and release
profile
Consistent quality.
1
2
High purity.

Purity & Consistent quality
Lipids developed with the application in mind: 1,3-GDO
• Very high isomeric purity
leading to defined and
controlled behaviour
Commercially available sample
Our high purity 1,3-GDO
Our high purity 1,3 & 1,2-GDO
• HPLC purity > 99% 1,3-GDO
• Highly defined
isomeric mixture (1,3 & 1,2)

Troubleshooting throughout the years
Are all those painpoints such a game changer?
Convenient and reproducible behavior in formulation: particle characteristics, stability and release profile.
Consistent quality.
1
2
3
Good material characteristics: solubility, cristallinity, stability, flowability
 influence on drug product GMP manufacturing process
High purity.

Cristallinity & Stability
Lipids developed with the application in mind: DOPC
Benefits
• Crystalline material
• Enhanced stability: more
than 7 years at 25 °C / 60% rH
• Fast dissoluton rate
• Free flowing powder: easy
weighing & portioning
• Patented:
WO 2009/146779Crystalline DOPC
DMF type II
available
Amorphous DOPC

Lipids developed with the application in mind: (R,S)-DOTAP Cl
Cristallinity & Solubility…and definition
Crystalline DOTAP
Amorphous, wax-like DOTAP
DMF type II available

Solubility & Flowability
Lipids developed with the application in mind: DOPE
Outstanding solubility, easier handling in comparison to conventional, amorphous material
of wax-like consistency.
• Free flowing powder
• Fast and complete dissolution
• DOPE appeared as lumps, gel & foam
• limited solubility even after lyophilization
Powder DOPEWax-like DOPE

Agenda
34

Customized lipids
Enhanced liposome surface characteristics
PARTICLE SURFACE MODIFICATION
… through conjugated lipids with multiple functionalities
• Lipidic nanocarriers have demonstrated their suitability for the delivery of
different APIs.
• Certain needs remain still unmet
• STEALTH particles
• TARGETED particles
• Avoid accumulation in undesired locations, off-target side effects, toxicity
• Improve efficacy, reduce dose, improve economy of treatment

In-solution lipidation of peptides
 Functional groups in the side chain lead to by-products
 Incompatibility of solubility result in low yields
 Expensive product
 Hardly scalable process
 Not suitable for GMP production
R RR R R R R RR R R R
SPPS
Resin
+ //
Customized Lipids: Our technology
Solid phase synthesis
 Start with a lipidated amino acid
 Uses large excess of reagents
 Fast work-up simply by washing the resin
 Can be automated
 Cheaper products
 Scalable process
 Suitable for GMP production
many functionalties are present in
the side chains of the targeting peptide
aqueous solubility aprotic solubility

Agenda
37

38
Where is our team located?
A global setup
Develop. & Launch Site
Schaffhausen
Global setup:
 Additional European site: Arklow (Ireland)
 US sites: St. Louis, Madison/Verona, Sheboygan (Wisconsin)
 Risk mitigation (multiple production sites)
Production Site
Altdorf
Buchs
Contract Manufacturing
We seek the needed
balance:
flexibility of
small teams
dedicated to Drug
Delivery
compounds and
the
global reach
with multiple
alternatives

A GMP manufacturing process that is scalable and reproducible in quality and yield facilitates
formulation and is a prerequisite for a consistent quality of the final product.
 We specialize in cGMP manufacturing that follows the concepts of ICH Q7
 Scale-up processes, from lab to kilo-lab to a production scale of up to 4,000 liters
The quality of lipids has a major impact on the performance of the liposomal formulation.
 We offer a consistently high product quality from grams to tons.
High regulatory requirements that are close to the ones for APIs.
 We support you with regulatory expertise and counsel from a dedicated team through all phases of
clinical development and commercialization
 Covering all aspects of quality assurance and documentation
Successful drug development with synthetic lipids
Summary
Our high purity lipids and our expertise in cGMP manufacture and regulatory matters
facilitate your formulation and support you from development to commercialization.

Judit Huarte Ciganda, PhD
judit.huarte-ciganda@emdgroup.com
The Vibrant M is a trademark of Merck KGaA, Darmstadt, Germany or its affiliates. All other trademarks are
the property of their respective owners. Detailed information on trademarks is available via publicly
accessible resources. © 2018 Merck KGaA, Darmstadt, Germany and/or its affiliates. All Rights Reserved.

Critical Aspects of Synthetic Lipids Drug Development

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