Note: Descriptions are shown in the official language in which they were submitted.
CA 022~328~ 1998-10-29
.
FIL~.~ ~'' ' ~. ' r
T~
S018-582J.200
Pharmaceutical Preparation in the form of Liposomes
The present invention relates to a pharmaceutical
preparation in the form of liposomes, suitable for the
solubilisation of poorly-soluble medicaments, especially
poorly-soluble neurokinine antagonists such as e.g. (+)-
camphor-3-carbonyl-(2S,4R)-4-hydroxypyrolyl(2S)-2-
naphthylalanyl-(2-methoxyphenyl)piperazide [BIIC 1996 BS]
or l-methylindol-3-yl-carbonyl-[4(R)-hydroxy]-L-prolyl-[3-
(2-naphthyl)]-L-alanine-N-benzyl-N-methylamide [FK 888].
Liposomes are generally ball-shaped structures - with a
diameter of 25 nm up to several ~m - comprising one or
more concentric lipid double layers with an aqueous
interior. This kind of lipid vesicle can be created by
mechanical fine distribution of phospholipids (e.g.
lecithin) in aqueous media. They do not only serve as
membrane models in biochemistry or molecular biology but
can also be used as carriers for medicaments.
A great number of active ingredients, which can be
incorporated in liposomes or can be absorbed by liposomes,
are known from the prior art. These kind of liposomal
systems are always preferred when the active ingredient is
to be released over a longer period of time, undesired
medicament effects are to be reduced, the active
ingredient is to be stabilised or liposomes are to serve
as target-selective medicament vehicles [P. Tyle and B.P.
Ram, Targeted Therapeutic Systems, Marcel Dekker Inc., New
York, 1990; G. Gregoriadis, Liposome Technology Vol. III,
Targeted Drug Delivery and Biological Interaction, CRC
Press Inc., Boca Raton, 1984]. The above mentioned
objectives were hitherto most adequately achieved by
enclosing water soluble medicaments in liposomes [M.J.
CA 022~328~ 1998-10-29
.,
Ostro, Liposome- From Biophysics to Therapeutics, Marcel
Dekker Inc., New York, 1987; G. Gregoriadis, Liposome
Technology Vol. II, Incorporation of Drugs, Proteins and
Genetic Material, CRC Press Inc., Boca Raton 1984].
In recent times, the aspect of medicament solubilisation
using liposomes has gained increasing significance.
Within the framework of preliminary research work, the
regularity of the inclusion of the liposomal medicament
has, however, remained hitherto relatively unresearched.
In general, the objective of the medicament solubilisation
is to have the medicament present in a mainly aqueous
formulation with a dispersed-molecular distribution. This
formulation can then be administered using known
application methods.
The following criteria should be met by medicament
preparations solubilised by liposomes:
The medicament should be included in liposome bilayers in
dispersed-molecular form with the proviso that the ratio
of included medicament to utilised quantity of lipids is
as high as possible. In the same way, the medicament
liposomes should have an active ingredient and stability
range which is adapted to the intended purpose.
For many application forms it is indispensable to be able
to produce sterile liposome preparations, for example by
sterilisation filtration with membrane filters with 0.2 ~m
pore diameter. Small liposomes, i.e. of liposome diameter
~200 nm are ideal for attaining satisfactory liposome
stability during mechanical agitation. This, for example,
is required during atomisation of aqueous liposome
preparations by means of pneumatic jet atomisers for
inhalative application. In order to avoid embolic
infusion incidents, it must be guaranteed that the size of
CA 022~328~ 1998-10-29
__
the injected liposomes on the one hand does not exceed a
critical value, and on the other hand the liposomes must
not form any aggregates of this critical size after
intravenous administration. Only physiologically-harmless
adjuvant substances are to be used as solubilising agents
for use with patients in general, and for intravenous
application in particular. The liposome components which
have been suggested by way of example are suitable as
natural bodily components, since for example on the one
hand they are not toxic and on the other they have a good
compatibility, and can be decomposed by the body without
leaving a residue.
The objective of the present invention is henceforth to
provide a medicament formulation on the basis of liposomes
for the solubilisation of poorly-soluble medicaments,
especially for neurokinine (tachykinine) antagonists, as
disclosed in the general formula, the preferred ranges and
in the embodiment examples of WO 95/30687. A principal
objective of the present invention is to provide a
medicament formulation on the basis of liposomes for the
solubilisation of the active ingredients (+)-camphor-3-
carbonyl-(2S,4R)-4-hydroxyprolyl-(2S)-2-naphthylalanyl-(2-
methoxy-phenyl)piperazide [BIIC 1996 BS] and 1-
methylindol-3-yl-carbonyl-[4(R)-hydroxy]-L-prolyl-[3-(2-
naphthyl)]-L-alanine-N-benzyl-N-methylamide [FK 888] (cf.
Ann. Drug. Data Rep. 15 (1993) 252). In addition,
liposomal inclusion compounds with the suggested compounds
according to the invention can also be produced with
corticoids - such as e.g. flunisolide hemihydrate or
beclomethasone dipropionate.
BIIC 1996 is a non-selective neurokinine antagonist which
binds to the NK1, NK2 and NK3 receptor with affinities in
the nanomolar range. Surprisingly, it was discovered that
the liposome formulations, according to the invention, for
CA 022~328~ 1998-10-29
the solubilisation of pharmaceutical active ingredients,
which are produced with the liposome components described
in the following text, have special advantages. The
liposome preparations which contain active substances
according to the invention, hence solve the objective as
stated above and also represent considerable progress in
the solubilisation of poorly water-soluble lipophile
medicaments.
The liposome compositions, according to the invention,
distinguish themselves e.g. in the way that liposome-
forming amphiphilic adjuvant substances, in which the
physico-chemical character of the hydrophilic molecular
portion is determined by glycerol or inositol, are
especially suitable for incorporating large quantities of
poorly-soluble medicament especially the neurokinine
antagonists BIIC 1996 BS and FK 888, as mentioned above -
in liposomes.
According to the invention, the liposome-forming adjuvant
substances correspond to general formulas I and II of
diagram 1, in which R1 and R2 in both general formulas,
independently from one another, have a branched or
unbranched alkyl group of a naturally, semi or fully
synthetically producable fatty acid - of saturated or
unsaturated character - with 1-30 carbon atoms.
Examples for saturated unbranched fatty acids are: formic
acid, acetic acid, propionic acid, butyric acid, valeric
acid, hexanoic acid, oenanthic acid, caprylic acid,
pelargonic acid, capric acid, undecanoic acid, lauric
acid, tridecanoic acid, myristic acid, pentadecanoic acid,
palmitic acid, margaric acid, stearic acid, nonadecanoic
acid, arachic acid, behenic acid, lignoceric acid, cerotic
acid, melissic acid.
CA 022~328~ 1998-10-29
The following are an example of saturated branched fatty
acids: isobutyric acid, isovaleric acid, tubercolostearic
acid.
The following are examples of mono-unsaturated unbranched
fatty acids: acrylic acid, crotonic acid, palmitoleic
acid, oleic acid, erucic acid.
The following are examples of di-saturated unbranched
fatty acids: sorbic and linoleic acid.
Examples of tri-saturated unbranched fatty acids are
linolenic acid and elaostearic acid.
Examples for tetra- and penta-unsaturated unbranched fatty
acids are arachidonic acid and clupanodonic acid
respectively.
Docosahexanoic acid is an example of a hexa-unsaturated
unbranched fatty acid.
The use of phosphatidylglycerols and phosphatidylinositols
in the form of the free acids or their salts - especially
in the form of the alkali salts - is preferred; use of the
sodium salts is especially preferred.
The use of the following phosphatidylglycerols is
especially preferred:
sodium salts of dimyristoylphosphatidylglycerol (DMPG-Na),
dipalmitoylphosphatidylglycerol (DPPG-Na) and
distearoylphosphatidylglycerol (DSPG-Na).
Use of the following compounds as phosphatidylinositols
are especially preferred:
CA 022~328~ 1998-10-29
sodium salts of dimyristolphosphatidylinositol (DMPI-Na),
dipalmitoylphosphatidylinositol (DPPI-Na) and
distearoylphosphatidylinositol (DSPI-Na).
For the purposes of clarification of the structural
composition of phosphatidylglycerols and
phosphatidylinositols, Fig. 1 shows especially the
structural formula of the sodium salt of
phosphatidylglycerols and phosphatidylinosltols.
The liposome components according to the invention are
characterised in that they can be added to any other
liposome components in proportions of greater than 0 mol.
~ up to 100 mol. ~.
The liposomes produced by means of the liposome component
parts according to the invention - especially BIIC 1996-
containing liposomes - have the following advantages:
1. The content of pharmaceutically-active components can
be strongly increased by addition of the liposome
components according to the invention. For this
reason, a high active ingredient concentration can be
obtained by dispensing with the cost-intensive
concentration increase of the remaining liposome
constituents.
2. By variation of the molar proportion of liposome
components according to the invention, the maximum
inclusion of the medicament can be controlled over a
wide area.
3. The liposome components according to the invention are
indispensable for the simple production of liposomes
which contain active ingredient in a size range of
<200 nm.
CA 022~328~ 1998-10-29
4. The concentration of the solubilised medicament can be
controlled by variation of the total lipid content.
5. The medicament enclosed in the liposome - especially
the neurokinine antagonist BIIC 1996 BS - stabilises
the liposomes with regard to their increase in size.
6. Liposomes with sub-maximum quantities of enclosed
medicament can be stabilised with regard to their size
increase by the addition of cholesterol.
7. Medicament containing liposomes show no tendency to
release enclosed medicaments either in the case of
mechanical agitation or extended storage.
In the following text, the term "lipid" or "lipids" refers
to all component parts of the liposome bilayers with the
exception of the medicament enclosed in the liposome.
The liposomes, comprising the liposome components
suggested according to the invention, can be produced
according to various processes. The following examples
explain the processes for the manufacture of liposomes
with lipophilic medicaments for the lipid compositions
according to the invention, and are known per se from the
prior art. [M. J. Ostro, Liposomes, Marcel Dekker, New
York, 1983; G. Gregroiadis, Liposome Technology Vol. I,
Preparation of Liposomes, CRC Press Inc., Boca Raton,
1984]:
Mechanical Methods
The hydration of the lipid medicament mixture extracted
from organic media, or hydration of lipids and medicament
CA 022~328~ 1998-10-29
without prior "film formation" from an organic solution,
is carried out using aqueous dispersion means.
The liposomes which result from spontaneous vesicle
formation during hydration can be varied in their size.
This dispersion can take place by mechanical energy input
of various types - for example, simple shaking by hand,
the use of agitation apparatus, by extrusion through a
membrane filter with exactly-defined pore size, extrusion
under high pressure through a variety of narrow jets, by
the use of a variety of agitation and mixing appliances
and by subjecting to ultrasonic treatment.
Removal of Solvent
Removal of an organic solvent from W/O emulsions,
comprising organic solvents, water, lipids and medicament.
Injection Method
Injection of organic lipid-medicament solutions in aqueous
dispersion means, with or without subsequent removal of
the organic solvent.
Detergent Method
Detergent removal from lipid-medicament-detergent
miscellae, for example via dialysis.
Freeze-Dry Method
Liposomes containing the active ingredient result from
spontaneous vesicle formation of freeze-dried lipid-
medicament mixtures during hydration. Liposomes
containing the medicament are reconstituted during the
hydration of freeze-dried liposomes.
CA 022~328~ 1998-10-29
Freeze-Thaw Method
Repeated freezing and thawing of aqueous lipid-medicament
systems results in the formation of liposome dispersions.
Individual application forms of the liposome formulations
according to the invention, are listed in the following
text without limiting the variety of possibilities for
using the system according to the invention. Liposomes
produced with the liposome components, according to the
invention, can be administered p.o., i.v., s.c., dermally,
intrapulmonary, nasally, i.p., rectally, i.m., ocularly or
intercerebrally.
The described invention will be explained again in the
following Example. Different formations will be apparent
to the skilled person from the present description.
However, it should be expressly noted that the Examples
and the description are only provided for the purposes of
explanation and are not to be seen as a restriction of the
invention.
CA 022~328~ 1998-10-29
.~_
- 10 -
EXAMPLES
Foreword:
General production methods for the following Examples:
The lipids in question were weighed in a round flask with
a long neck and were dissolved in a suitable mixture of
chloroform and methanol, optionally using heat.
Medicament dissolved in methanol was added by pipette.
The solvent mixture was evaporated down in a rotary
evaporator and dried. The lipid film was re-dried in a
vacuum oven for complete removal of residual solvent.
After degassing with nitrogen, the lipid film was stored
at -18~C until it was hydrated. In order to produce the
liposomes, lipid film was hydrated with glucose solution
5~ (m/V). Hydration was carried out for approximately 4
hours at 75~C, wherein approximately 10-13 glass balls
with a diameter of 6 mm were added to the mixture to
promote easier dispersion and hydration of the lipid-
medicament mixture. From time to time, the round flask
was agitated strongly by hand during hydration. The
resulting liposomes were subjected to ultrasound treatment
to reduce their size. For this reason, the liposome
dispersion was decanted into thick-walled acoustic
irradiation glasses and was irradiated with the Branson
Sonifier B15 Cell Disruptor. The following apparatus
parameters were selected for the standard mixture size of
10 ml: sonic rod 1/2~'; irradiation time 20 minutes, type of
irradiation pulsed; duty cycle 70~; output control 8;
temperature bath 50~C. In order to remove metal dust,
larger liposomes and medicament which was not enclosed in
the liposomes, the cooled preparation was firstly
centrifuged for 15 minutes at 1900 g, and following this
the supernatent was pressed through a membrane filter with
a pore width of 0.2 ~m. The liposome size was determined
CA 022~328~ 1998-10-29
- 11 -
in the filtrate and the preparation was stored for 24
hours at 4~C and 24 hours at room temperature. The
average liposome size was always below 100 nm. After
repeated filtration through a membrane filter with a pore
width of 0.2 ~m, the liposome size, the medicament content
and the lipid content were determined in filtrate. For
tests over the storage time, the batches were divided into
two and following "filtration through a bacteria-retaining
filter" (DAB 10) were stored at temperatures 4~C and room
temperature with the exclusion of light.
Table 1
Abbreviations of the utilised lipids
Abbreviation Lipid
DSPG-Na Distearoylphosphatidylglycerol,
Na-salt
HSPC Hydrated soya-phosphatidylcholine
(Trade name: Phospholipon~ 100 H,
Company Nattermann, Cologne (D))
CH Cholesterol
PI-Na Phosphatidylinositol, Na-salt
DPPS Dipalmitolylphosphatidylserine
25 DPPA Dipalmitolylphosphatidylic acid
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,._~ ,_
- 12 -
Example 1
The influence of various lipids on the liposomal inclusion
of BIIC 1996 BS
The four lipids DSPG-Na, PI-Na, DPPS and DPPA were tested
in a combination with 30 mol. ~ HSPC for their inclusion
capacity with regard to BIIC 1996 BS. A lipid
concentration of 5 ~mol/ml and a medicament concentration
of 10.2 mg/ml were used. Diagram 2 shows the
corresponding result, wherein only the glycerol and
inositol-containing lipids showed a satisfactory result.
The quotient D/L represents the molar ratio of BIIC 1996
BS contained in the liposomes and lipid.
Example 2
Influence of phosphatidylglycerol on the liposomal
inclusion of BIIC 1996 BS
HSPC/DSPG-Na in varying compositions as a lipid mixture
was tested.
Table 2 summarises the tested lipid mixtures:
CA 022~328~ l998-l0-29
- 13 -
Table 2
Tested lipid mixtures
5 HSPC DSPG-Na CH
[mol-~] [mol-~] [mol-~]
0
1050 50 0
0
0 100 0
49 21 30
1521 49 30
0 70 30
0 85 15
0 55 45
0 40 60
2027.75 64.75 7.5
25. 5 59.5 15
16.5 38.5 45
12 28 60
25 The more DSPG-Na added to the various lipid mixtures, the
more active ingredient (BIIC 1996 BS) could be included in
the liposomes. No preparations with liposomes of <200 nm
could be produced with the considered lipid concentration
of 45 ~mol/ml with the applied production methods without
30 DSPG-Na addition.
Figures 3 - 6 show the results in graphic form.
CA 022~328~ 1998-10-29
Example 3
Influence of lipid concentration on the liposomal
inclusion of BIIC 1996 BS.
The lipid concentrations of 0 ~mol/ml to 75 ~mol/ml were
varied for the lipid compositions HSPC/DSPG-Na/CH
(21/49/30) and HSPC/DSPG-Na (30/70). Diagram 7 shows that
the quantity of BIC 1996 BS increases with increasing
lipid concentration, and hence the medicament content can
be controlled.
Example 4
Influence of the applied concentration of medicament on
the liposomal inclusion of BIIC 1996 BS.
The medicament concentrations were varied between 0 mg/ml
to 10.2 mg/ml for the lipid compositions HSPC/DSPG-Na
(30/70) and HSPC/DSPG-Na/CH (21/49/30).
Figures 8 and 9 show the results in graphic form; they
show that the quantity of liposomally-included medicament
increases in a linear fashion until it reaches the
saturation threshold with increasing quantities of applied
medicament. In this way, the quantity of included BIIC
1996 BS can be controlled in an excellent manner.
Example 5
Influence of storage conditions and applied concentration
on the medicament content of BIIC 1996 BS liposomes
The medicament concentrations were varied between 0 mg/ml
to 10.2 mg/ml, for the lipid compositions HSPC/DSPG-Na
(30/70) and HSPC/DSPG-Na/Cl (21/49/30), and the inclusion
CA 022~328~ l998-l0-29
." . ._
- 15 -
rates were tested at 2 days after production and 28 days
after production. The storage conditions were 4~C and
room temperature (19-24~C), with light being excluded.
Figures 10 and 11 represent the results in graphic form,
5 these show that the medicament inclusion did not
significantly alter during the period of the test.
Example 6
Influence of storage conditions and applied concentration
on the size of BIIC 1996 BS liposomes
The medicament concentrations were varied between O mg/ml
and 10. 2 mg/ml for the lipid compositions HSPC/DSPG-Na
(30 / 70) and HSPC/DSPG-Na/CH (21 / 49 / 30), and the liposome
size was tested after production, 2 days after production
and 28 days after production. The storage conditions were
4~C and room temperature (19-24~C), light being excluded.
20 Increasing quantities of BIIC 1996 BS stabilised the
liposome size (Fig. 12) . An addition of 30 mol~
cholesterol stabilised liposomes with less than the
maximum inclusion of medicament with regard to the
increase in size (Fig. 13) .
Example 7
Stability of BIIC 1996 BS liposomes
30 Liposome dispersions of lipid composition HSPC/DSPG-Na
(30/70) with the medicament concentration 3.5 mg/ml and
HSPC/DSPG-Na/CH (21/49/30) with the medicament
concentration 3.0 mg/ml were atomised with a lipid
concentration of 45 ~mol/ml with the Respirgard II ~ 10-
35 atomising system. The jet gas flow was 6 l/min,atomisation duration was 22.53 min + 2.13 min (x + SD).
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- 16 -
Before and after the atomising, the atomiser solution in
the reservoir was tested for medicament content. Captured
aerosol was also tested by means of the Andersen 1-AFCM
Non-Viable Ambient Particle Sizing Sampler Mark II, driven
by water steam-saturated air. Fig. 14 clarifies the high
content stability of the examined liposomes in the case of
very strong mechanical load, as occurs during jet
atomising.
Explanation of the Diagrams
Fig. 1: shows the structural composition of
phosphatidylglycerols and phosphatidyl-inositols -
especially the structural formula of the sodium salt of
phosphatidyl- glycerols and phosphatidylinositols [G.
Cevc, D. Marsh, Phospholipid Bilayers, Physikal Principles
and Models, John Wiley & Sons Inc., New York, 1987; D.
Arndt, I. Fichtner, Liposomen, Darstellung - Eigenschaften
- Anwendung, Akademie Verlag Berlin 1986].
Fig. 2: influence on various lipids on the liposomal
inclusion of BIIC 1996 BS (Example 1).
Inclusion rates 2 days after production.
Lipid composition: HSPC/Lipid (30/70)
Applied concentration: BIIC 1996 BS >10 mg/ml
Lipid: 5 ~mol/ml
The data shown refers to the arithmetic mean value from 3
tests.
The error bar represents the standard deviation.
Fig. 3: shows the influence of phosphatidylglycerol on
the liposomal inclusion of BIIC 1996 BS (Example 2).
Inclusion rates 2 days after production.
CA 022~328~ l998-l0-29
,_~
- 17 -
Utilised concentrations: BIIC 1996 BS > 10 mg/ml
Lipid 45 ~mol/ml
The data shown refers to the arithmetic mean value from 3
tests.
The error bar represents the standard deviation.
Fig. 4: shows the influence of phosphatidylglycerol on
the liposomal inclusion of BIIC 1996 BS in three component
lipid systems with constant cholesterol proportions
(Example 2).
Inclusion rates 2 days after manufacture.
Utilised concentrations: BIIC 1996 BS > 10 mg/ml
Lipid 45 ~mol/ml
The data shown refers to the arithmetic mean value from 3
tests.
The error bar represents the standard deviation.
Fig. 5: shows the influence of phosphatidylglycerol on
the liposomal inclusion of BIIC 1996 BS in two component
lipid systems with cholesterol (Example 2).
(-) D/L = D/DSPG-Na
(~) D/L = D/(DSPG-Na + CH)
Inclusion rates 2 days after production.
Utilised concentrations: BIIC 1996 BS > 10 mg/ml
Lipid 45 ~mol/ml
The data shown refers to the arithmetic mean value from 3
tests.
The error bar represents the standard deviation.
CA 022~328~ 1998-10-29
- 18 -
Fig. 6: shows the influence of phosphatidylglycerol on
the liposomal inclusion of BIIC 1996 BS in three component
systems with variable cholesterol proportions (Example 2).
(-) D/L = D/HSPC-Na
(~) D/L = D/(HSPC-Na + DSPG-Na + CH)
Inclusion rates 2 days after production.
Utilised concentrations: BIIC 1996 BS ~ 10 mg/ml
Lipid 45 ~mol/ml
The data shown refers to the arithmetic mean value from 3
tests.
The error bar represents the standard deviation.
Fig. 7: shows the influence of the lipid concentration on
the liposomal inclusion of BIIC 1996 BS (Example 3).
(-) HSPC/DSPG-Na (30/70)
(~) HSPC/DSPG-Na/CH (21/49/30)
Inclusion rates 2 days after production.
Utilised concentrations: BIIC 1996 BS > 10 mg/ml.
The data shown refers to the arithmetic mean value from 3
tests.
The error bar represents the standard deviation.
Fig. 8: shows the influence of the utilised concentration
of the medicament on the liposomal inclusion of BIIC 1996
BS (Example 4).
Inclusion rates 2 days after production.
Lipid composition: HSPC/DSPG-Na (30/70)
Utilised concentration: Lipid 45 ~mol/ml
CA 022~328~ l998-l0-29
- 19 -
The data shown refers to the arithmetic mean value from 3
tests.
The error bar represents the standard deviation.
5 Fig. 9: shows the influence of the utilised
concentrations of the medicament on the liposomal
inclusion of BIIC 1996 BS (Example 4) .
Inclusion rates 2 days after production.
Lipid composition: HSPC/DSPG-Na/CH (21/49/30)
Utilised concentrations: Lipid 45 ~mol/ml
The data shown refers to the arithmetic mean value from 3
tests.
15 The error bar represents the standard deviation.
Fig. 10: shows the influence of the utilised
concentrations of the medicament on the liposomal
inclusion of BIIC 1996 BS (Example 5) .
Inclusion rates 2 days after production and 28 days after
storage at 4~C and room temperature, light being excluded.
Lipid composition: HSPC/DSPG-Na/CH (30/70)
Utilised concentrations: Lipid 45 ~mol/ml
The data shown refers to the arithmetic mean value from 3
tests.
The error bar represents the standard deviation.
30 Fig. 11: shows the influence of storage conditions and
utilised concentration of the medicament on the liposomal
inclusion of BIIC 1996 BS (Example 5) .
Inclusion rates 2 days after production and 28 days after
35 storage at 4~C and room temperature, light being excluded.
Lipid composition: HSPC/DSPG-Na/CH (21/49/30)
CA 022~328~ 1998-10-29
_ _
- 20 -
Utilised concentrations: Lipid 45 ~mol/ml
The data shown refers to the arithmetic mean value from 3
tests.
The error bar represents the standard deviation.
Fig. 12: shows the influence of storage conditions and
utilised concentrations of the medicament on the size of
BIIC 1996 BS liposomes (Example 6).
Liposome sizes after production (-), after 2 days (~),
after 28 days, after storage at 4~C (-) and after 28 days
storage at room temperature (19-24~C) (O), light being
excluded.
Lipid composition: HSPC/DSPG-Na (30/70)
Utilised concentrations: Lipid 45 ~mol/ml
The data shown refers to the arithmetic mean value from 3
tests.
The error bar represents the standard deviation.
Fig. 13: shows the influence of storage conditions and
utilised concentrations of the medicament on the size of
BIIC 1996 BS liposomes (Example 6).
Liposome sizes after production (-), after 2 days (~),
after 28 days, after storage at 4~C (-) and after 28 days
storage at room temperature (19-24~C) (O), light being
excluded.
Lipid composition: HSPC/DSPG-Na (21/49/30)
Utilised concentrations: Lipid 45 ~mol/ml
The data shown refers to the arithmetic mean value from 3
tests.
CA 022~328~ 1998-10-29
__
The error bar represents the standard deviation.
Fig. 14: shows the atomisation stability of BIIC 1996 BS
liposome dispersions (Example 7).
The liposome dispersions were atomised with the Respirgard
II ~ 10-System.
Utilised concentrations:
BIIC 1996 BS HPSC/DSPG-Na (30/70) 3.5 mg/ml
HSPC/DSPG-Na/CH (21/49/30) 3.0 mg/ml
Lipid concentration: 45 ~mol/ml
Jet gas flow 6 l/min; atomisation duration 22.53 + 2.13
min (x + SD).
The data shown refers to the arithmetic mean value from 5
tests.
The error bar represents the standard deviation.
