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Newsletter
Volume 4, Number 1
November 2009
Newsletter Volume 4, Number 1
November 2009
Page 2
Phospholipid Research Center News
Meeting of the Scientific Board, October 2, 2008
in Ludwigshafen
It should contain the following paragraphs:
1.
2.
3.
4.
5.
6.
7.
Introduction
Aim
Preliminary results
Conclusions
Future perspectives
Publications
References
Participants:
Prof. Alfred Blume (Scientific Board)
Prof. Gert Fricker (Scientific Board)
Dr. Frank Martin (Scientific Board)
Prof. Christel Müller-Goymann (Scientific Board)
Dr. Ralf-Olaf Quinkert (Scientific Board)
Dr. Herbert Rebmann (PRC)
Dr. Constanze Setzer (PRC)
Mr. Armin Wendel (PRC)
Dr. Jürgen Zirkel (PRC)
Announcement of new Managing Director
At the last meeting of the Scientific Board in
Ludwigshafen, it was decided to support the following submitted research proposals:
On October 1st 2009, Dr. Constanze Setzer took
over the position as Managing Director of the
Phospholipid Research Center.
For more information about the funding of projects,
please visit our website: www.phospholipids.net.
Martin Brandl, University of Southern Denmark:
“Oral bioavailability Screening of new Drug
Compounds: Comparison of the Phospholipid
Vesicle based Model with the Caco-2 Model”
(Supplemental funding)
To simplify the applications in future, a general
form was advised. The proposal (4-5 pages in
length) should be divided into the following parts:
1.
2.
3.
4.
5.
6.
7.
Abstract
Introduction to the topic
Aim of the project
Methods
Work plan
Timeline
Costs
Dr. Setzer holds a Diploma in Chemistry and a
Master in Business Administration. She has 15
years of management experience in Research and
Development and Technical Customer Service in
the chemical industry.
An annual report should reveal the progress of the
funded project.
Phospholipid
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Newsletter Volume 4, Number 1
November 2009
Page 3
Industry News
Lipid Therapeutics and Dr. Falk Pharma to develop a new treatment approach to ulcerative
colitis
Lipid Therapeutics GmbH and Dr. Falk Pharma
GmbH announced that they have entered into a
co-development and licensing agreement for product LT-02 applied in the treatment of ulcerative
colitis. Ulcerative colitis is a disabling inflammation
of the lower gut, affecting more than one million
people worldwide. Lipid Therapeutics’ LT-02 is a
specially formulated phospholipid that augments
the natural protective mucosal barrier in the lower
gut. With this treatment approach, Lipid Therapeutics has the first program worldwide that targets the
pathological changes in the lower gut barrier function of colitis patients, which are thought to be one
of the main underlying disease causes. This compelling mechanism of action, which has already
demonstrated activity in clinical trials, thus holds
great promise for the treatment of this disease.
Lipid Therapeutics and Dr. Falk Pharma will jointly
develop LT-02 through a phase IIb clinical trial.
MediGene reorganizes to focus on clinical programs
The German biotechnology company MediGene is
restructuring operations to focus on clinical R&D
programs. It will close its early-stage research
department and redeploy the staff and financial
resources to the company's clinical development
and manufacturing departments so the measures
will involve few redundancies. One of MediGene's
two clinical programs is for EndoTAG-1. EndoTAG1 is designed to destroy existing tumor blood vessels, targeting their negatively charged endothelial
cells with its cationic carrier liposomes. It is different from anti-angiogenic drugs, which block the
formation of new tumor blood vessels.
Phospholipid
Forschungszentrum e.V./
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The product is in Phase II testing for the treatment
of pancreatic cancer and breast cancer and in
Phase I for other oncology indications.
Transave's liposomal amikacin promising in
bronchiectasis
Transave currently develops an inhaled liposomal
formulation of the aminoglycoside antibiotic amikacin, Arikace. The US company has shown positive results in a Phase II trial in patients with bronchiectasis and pseudomonal lung infections. Amkacin has been widely available for a number of
years as an injectable agent for the treatment of
susceptible bacterial infections. There is currently
no antibiotic specifically approved for the treatment
of bronchiectasis or associated lung infections in
the US or EU.
Compilation of literature: Constanze Setzer
Literature Report
Formulation and Evaluation of Liposomes of
Ketoconazole
Patel R.P., Patel H., Baria A.H., International
Journal of Drug Delivery Technology 1(1):16-23,
2009.
Multilamellar vesicles for topical application
encapsulating the antifungal drug Ketoconazole
were prepared using soya lecithin. Liposomes
were prepared by thin film method and
characteristics as well as in vitro release were
studied. It was found that drug release from the
liposomes primarily depends on the lipid
composition, drug to lipid ratio and also the volume
of hydration medium. Possible advantages of
encapsulation of Ketoconazole into liposomes are
Im Neuenheimer Feld 582
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Newsletter Volume 4, Number 1
November 2009
Page 4
less side effects and prolonged release of the active.
Production of Lipid Microparticles Magnetically
Active by a Supercritical Fluid-based Process
embodiments, the pharmaceutically active
compounds are ansamycins and the overall
formulation is substantially devoid of medium and
long chain triglycerides.
Vezzù, K., Campolmi, C., Bertucco, A.,
International Journal of Chemical Engineering
2009:1-9.
US Pat. Appl. 20090238865; September 24, 2009
Magnetite nanoparticles were coated with mixtures
of phospholipids, triestearin and oleic acid to increase their bioavailability. The magnetite particles
are manufactured by a technique based on gas
saturated solution process working with supercritical CO2.The characteristics of the particles and
encapsulation of the magnetite nanoparticles were
studied. The process leads to small lipid coated
nanoparticles, which can be trapped by an external
magnet producing a magnetic field compatible with
human application.
The invention concerns nanocapsules, in particular
with an average size less than 50 nm, consisting of
an essentially lipid core liquid or semiliquid at room
temperature, coated with an essentially lipid film
solid at room temperature having a thickness of
2-10 nm. The invention also concerns a method for
preparing same which consists in producing a reverse phase of an aqueous emulsion brought
about by several temperature raising and lowering
cycles. Said lipid nanocapsules are particularly
designed for producing a medicine.
US Pat. Appl. 20090258065; October 15, 2009
US Pat. Appl. 20090226525; September 10, 2009
Dermatological
compositions
avermectin nanocapsules
Self-assembling nanoparticle drug delivery system
comprising
Compositions and nanoemulsions containing lipid
nanocapsules dispersed in a hydrophilic phase,
such nanocapsules including at least one
avermectin compound, are useful for the treatment
of dermatological pathologies, e.g., rosacea.
US Pat. Appl. 20090238880; September 24, 2009
Phospholipid-based pharmaceutical formulations
and methods for producing and using same
Pharmaceutical formulations and methods of
producing and using the same are described and
claimed. The formulations are dispersions of
phospholipids and one or more pharmacologically
active compounds, pharmaceutically acceptable
salts thereof, or prodrugs thereof. In specific
Phospholipid
Forschungszentrum e.V./
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Lipid nanocapsules, preparation process and use
as medicine
A self-assembling nanoparticle drug delivery
system for the delivery of various bioactive agents
including peptides, proteins, nucleic acids or
synthetic chemical drugs is provided. The
self-assembling nanoparticle drug delivery system
described herein includes viral capsid proteins,
such as Hepatitis B Virus core protein, encapsulating the bioactive agent, a lipid layer or
lipid/cholesterol layer coat and targeting or
facilitating molecules anchored in the lipid layer. A
method for construction of the self-assembling
nanoparticle drug delivery system is also provided.
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Newsletter Volume 4, Number 1
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US Pat. Appl. 20090226509; September 10, 2009
Composition
disorders
for
treatment
of
inflammatory
A pharmaceutical composition for parenteral
administration, comprising liposomes composed of
non-charged vesicle-forming lipids, optionally
including not more than five (5) mole percent of
charged vesicle-forming lipids, the liposomes
having a selected mean particle diameter in the
size range between about 40-200 nm and containing a water soluble corticosteroid for the
site-specific treatment of inflammatory disorders, is
provided.
Compilation of literature: Torsten Kromp
Conference Report
First Symposium on “Phospholipids in Pharmaceutical Research” on May 10-11, 2009 in
Heidelberg, Germany.
On May 10-11, 2009, the 1st Symposium on Phospholipids in Pharmaceutical Research took place at
the TP Conference Center Technologiepark Heidelberg, Germany.
The aim of the meeting was to provide a platform
for discussion and contact throughout the spectra
of interest of phospholipid scientists.
Following the address of welcome by Dr. Herbert
Rebmann, the Chairman of the Phospholipid Research Center, Prof. Christel Müller-Goymann,
Braunschweig, a member of the Scientific Board of
the Phospholipid Research Center and one of the
organizers, opened the scientific program. The
latter consisted of 15 presentations and more than
50 posters covering many different aspects such
as production and analysis of phospholipids, their
Phospholipid
Forschungszentrum e.V./
Research Center
physical properties and the experience with their
use in pharmaceutical products as useful additives.
The poster session took place in the morning of
the second day of the meeting.
Three oral presentations dealt with research subjects that were part of projects funded by the
Phospholipid Research Center. One, entitled: Tetraether Lipid based Liposomes as oral Drug Delivery System. “, was given by Prof. Gert Fricker,
Heidelberg. Prof. Natasa Skalko-Basnet, Tromsø
reported on the recent results of her research
study focused on “Phospholipid-based Delivery
Systems for Phytochemicals”. The third talk given
by Dr. Mona Tawab covered “Drug-PhospholipidComplexes: Their pharmaceutical Relevance and
structural Properties.
The Phospholipid Research Center presented one
poster of a funded project. It was together with
Prof. Fricker, Heidelberg, about the “Oral Bioavailability of pharmaceutical Actives by NanoSolve”.
In addition three posters were presented that introduced research projects funded by the Phospholipid Research Center. One poster was from Dr.
Peters, Freiburg, about the project “Phosphatidylcholines in Anticancer Drug Delivery: Mere innocent Bystanders?” Another introduced latest findings in the project “Oral Bioavailability Screening:
Comparison of the Phospholipid Vesicle-based
Permeation Barrier with Caco-2 Cells”. It was presented by Sarah Fischer, Heidelberg. The third
poster by Jan Hüsch dealt with “DrugPhospholipid-Complexes: Their pharmaceutical
Relevance and structural Properties.”
A reception as well as the conference dinner took
place in the Castle of Heidelberg. Italian as well as
regional delicacies were served.
The 130 participants from all over the world perceived the meeting as informative in a relaxed
Im Neuenheimer Feld 582
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Newsletter Volume 4, Number 1
November 2009
Page 6
atmosphere. A number of fruitful discussions took
place after the oral presentations as well as during
the poster session and the breaks. The pleasant
social program with the guided tour through the
Castle of Heidelberg was well received by the participants.
Lysopholipids occur in almost every biological
membrane in mammalians. In living cells, LPL
results from membrane phospholipids through the
enzymatic action of a phospholipase A2 (PLA2),
which cleaves the fatty acid from the 2-position of
the glycerol backbone.
In blood, LPL is generated by PLA2 or by the lecithin-cholesterol-acyl-transferase (LCAT) from
phospholipids present in lipoproteins. LCAT transfers a fatty acid from phosphatidylcholine (PC) to
cholesterol, which results in a cholesterolester and
lysophosphatidylcholine (LPC).i
The audience from all over the world during the lectures.
Constanze Setzer
Information about a special
Phospholipid
Lysophosphatidylcholine
Lysophospholipids (LPLs) are phospholipids that
are missing one of their two O-acyl chains. They
have a 3-carbon glycerol (or –sphingoid) backbone
on which a single carbonyl chain of varied length
and saturation is attached. One prominent LPL is
lysophosphatidylcholine (LPC).
As already mentioned, lysophosphatidylcholine is a
naturally occurring component in lecithin and
phospholipids. It is produced by enzymatic cleavage of phosphatidylcholine by phospholipase A2.
Furthermore, it can result from spontaneous hydrolysis of phosphatidylcholine during manufacturing or storage.
Pure lysophosphatidylcholine is a white powder
that is soluble in ethanol and dispersable in water.
Compared to phosphatidylcholine, lysophosphatidylcholine has increased solubility in water and, in
blends with other phospholipids, greater emulsifying activity for the formation of oil-in-water emulsions.
Lysophosphatidylcholine, like LPLs in general, can
affect fundamental cellular functions, i. e. proliferation, differentiation, survival, migration, adhesion,
invasion and morphogenesis. These functions
influence many biological processes that include
neurogenesis, angiogenesis, wound healing, immunity and carcinogenesis. It accumulates in pathological tissues e. g. in the ischemic myocardium
and artheriosclerotic aortas and is participating in
inflammation processes.
Recently, a novel lipid complex to improve oral
delivery of drugs was developed. The lipid com-
Phospholipid
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Newsletter Volume 4, Number 1
November 2009
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plex, which is called LYM-X-SORB, is a highly
organized lipid matrix which consists of lysophosphatidylcholine, monoglyceride and fatty acid. It
was shown to improve the oral delivery of Fenretinide (N-(4-hydroxyphenyl) retinamide) in mice up
to 4-fold in plasma and up to 7-fold in tissue, compared to conventional delivery.
Lysophosphatidylcholine, like LPLs in general, was
also proven as adsorption enhancers for nasal
administration of drugs.
The Food and Drugs Administration (FDA) in the
USA classified lysophosphatidylcholine and LPLs
in general as GRAS (generally recognized as safe)
in human food.
LPLs were tested for skin tolerance and it was
concluded, that they can be declared of safe as
cosmetic agent
Constanze Setzer
Analytical Column
HPLC in Phospholipid Analytics
HPLC (High Performance Liquid Chromatography)
is a widely used technique for separation of chemical substances from nearly every field of chemistry,
pharmaceutics and life sciences. Although there
are many preparative applications of liquid chromatography, which are also relevant for phospholipids, in this column the analytical side of HPLC will
be illustrated.
Basically, there are two different phase systems in
liquid chromatography, the normal and the reversed phase system. In normal phase systems
the packing material of the column, the so-called
stationary phase, consists of highly porous material
with a polar surface, usually a silica gel. The secPhospholipid
Forschungszentrum e.V./
Research Center
ond part of the system, the liquid phase, consists
of a non polar solvent with variable amounts of a
polar modifier. Depending on the polarity of the
compounds to separate, typical mobile phases are
hexane/ethyl acetate, hexane/2-propanol, chloroform/methanol etc. The retention time of a specific
analyte is now dependent form the partition of the
analyte between the stationary and the mobile
phase. The more polar the analyte, the longer is
the retention time.
The reversed phase system works by the same
principles, though the phases show opposite behavior with respect to polarity of the analytes. As
stationary phase usually a silica gel is used, which
surface is modified by chemically bonding a nonpolar moiety. In most cases a C8 or C18 hydrocarbon chain is used. Such a modified surface is
highly hydrophobic in contrast to the hydrophilic
surface of the unmodified silica gel. The mobile
phase consists of a polar component, usually water and an organic modifier, often methanol or acetonitril. The higher the content of the organic modifier, the shorter is the retention time of the analyte.
In the case of phospholipids these two different
phase systems allow for the performance of two
different separation tasks. By using the normal
phase set up, separation behavior is dominated by
the polarity of the phospholipids. Hence, the polar
head group of a phospholipid has the major impact
on the retention characteristics. The non polar
hydrocarbon chains of the fatty acids, bound to the
polar head group, show nearly no effect on retention time. This is a big advantage for the analysis
of phospholipids from natural sources. An example
is the characterization of the PC content in lecithin,
which requires a method that separates PC from
PE, lyso-PC, Sphingomyelin etc. but does not
separate the PC species that differ in the fatty acid
pattern only.
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Newsletter Volume 4, Number 1
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Analyzing phospholipids under reversed phase
conditions show totally different chromatograms.
The polar head group is also important for the retention behavior. However, in this case the nonpolar hydrocarbon chains of the fatty acids show
an important effect on retention. Apart from the
length of the hydrocarbon chains, the number of
double bonds has an impact on the retention time
of a specific phospholipid. Hence, HPLC analysis
of phospholipids from natural sources shows rather
complex chromatograms with many overlapping
peaks.
Besides the phase system, the choice of the detector plays an important role in HPLC analysis of
phospholipids. The most commonly used detector
in HPLC is the UV or UV-VIS detector. It uses the
absorption characteristics of the analyte at a specific wavelength. To select an optimal wavelength
two aspects have to be considered. Firstly, the UVor UV-VIS spectrum of the analyte should show an
absorption band with a high absorption coefficient.
Secondly, the mobile phase should exhibit absorption as low as possible at this specific wavelength.
This implies for phospholipids that UV detection is
limited to the region below 230 nm and to mobile
phases such as water, acetonitrile and methanol
respectively, which are usually applied under reversed phase conditions. Additionally, due to low
absorption coefficients, the sensitivity of UV detection is rather low for phospholipids.
ammonia, tri-ethylamine etc. The second requirement can be met easily as phospholipids have
sufficiently high molecular masses. Consequently,
the ELSD should be the ideal detector for phospholipids, however, there are some disadvantages
too. Although the sensitivity is better than for UV
detectors, it is often difficult to detect small
amounts of by-products with a concentration of
less than 0.5 %. In this cases thin layer chromatography is still the method of choice, because it is
reliable and fast. A second disadvantage is due to
the measuring principle. Especially for low concentrations, there is no linear relation between the
amount of analyte and the detector signal.
A third detector with a fast growing importance is
the mass spectrometer. Since the development of
efficient coupling techniques between HPLC and
mass spectrometry it becomes more and more
important, especially for the detection of trace
amounts of contaminants and for structure determination. In one of the next “analytical columns” a
more detailed review on phospholipids and mass
spectroscopy will be provided.
Ralf-Olaf Quinkert
An often used alternative to the UV-detector for the
detection of Phospholipds and other substance
with low UV absorption is the Evaporative Light
Scattering Detector (ELSD). Principle requirements
are a mobile phase which is completely volatile
and an analyte which is not volatile. The first requirement excludes the use of phosphoric acid and
its salts for adjusting pH, but in most cases it can
be substituted by other buffering substances such
as acetic acid, formic acid, tri-fluoro acetic acid,
Phospholipid
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Newsletter Volume 4, Number 1
November 2009
Page 9
Contact
Phospholipid Research Center
Im Neuenheimer Feld 582
69120 Heidelberg
Germany
Phone: +49 (0)6221 / 588 8360
Fax:
+49 (0)6221 / 651 5665
E-Mail: [email protected]
Web:
www.phospholipids.net
Disclaimer
This newsletter is provided “as is” and without warranty,
express or implied. All warranties with regard to the accuracy, reliability, timeliness, usefulness or completeness of the
information contained in the newsletter are expressly disclaimed. All implied warranties of merchantability and fitness
for a particular use are herby excluded. None of the information provided in the newsletter constitutes, directly or indirectly, the practice of medicine, the dispensing of medical
services, the recommendation to buy or use a product.
External links are provided in the newsletter solely as a
convenience and not as an endorsement of the content on
such third-party websites. The Phospholipid Research Center
is not responsible for the content of linked third-party
websites and does not make any representations, warranties
or covenants regarding the content or accuracy of materials
on such third-party websites. If you decide to access linked
third-party websites, you do so at your own risk.
Phospholipid
Forschungszentrum e.V./
Research Center
Im Neuenheimer Feld 582
D-69120 Heidelberg
VR 3192
Phone +49 (0) 6221 / 588 8360
Fax +49 (0) 6221 / 651 5665
www.phospholipids.net