U.S. patent application number 17/350363 was filed with the patent office on 2021-10-07 for controlled release peptide formulations.
This patent application is currently assigned to CAMURUS AB. The applicant listed for this patent is CAMURUS AB. Invention is credited to Markus Johnsson, Catalin Nistor, Fredrik Tiberg.
Application Number | 20210308226 17/350363 |
Document ID | / |
Family ID | 1000005657212 |
Filed Date | 2021-10-07 |
United States Patent
Application |
20210308226 |
Kind Code |
A1 |
Tiberg; Fredrik ; et
al. |
October 7, 2021 |
CONTROLLED RELEASE PEPTIDE FORMULATIONS
Abstract
The present invention relates to compositions forming a low
viscosity mixture of: TABLE-US-00001 a) 20-80 wt. % of at least one
diacyl glycerol and/or a tocopherol; b) 20-80 wt. % of at least one
phosphatidyl choline (PC); c) 5-20 wt. % of at least one
biocompatible, organic mono-alcoholic solvent; d) up to 20 wt. %
polar solvent e) at least one peptide active agent; f) optionally
at least one antioxidant; wherein the ratio of components a:b is in
the range 40:60 to 54:46; wherein the pre-formulation forms, or is
capable of forming, at least one liquid crystalline phase structure
upon contact with excess aqueous fluid. The invention further
relates to methods of treatment comprising administration of such
compositions, and to pre-filled administration devices and kits
containing the formulations.
Inventors: |
Tiberg; Fredrik; (Lund,
SE) ; Nistor; Catalin; (Lund, SE) ; Johnsson;
Markus; (Lund, SE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
CAMURUS AB |
Lund |
|
SE |
|
|
Assignee: |
CAMURUS AB
Lund
SE
|
Family ID: |
1000005657212 |
Appl. No.: |
17/350363 |
Filed: |
June 17, 2021 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
14117994 |
Feb 20, 2014 |
|
|
|
PCT/EP2012/059917 |
May 25, 2012 |
|
|
|
17350363 |
|
|
|
|
61489886 |
May 25, 2011 |
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 47/10 20130101;
A61K 47/14 20130101; A61K 38/31 20130101; A61K 9/0019 20130101;
A61K 38/00 20130101; A61K 9/0024 20130101; A61K 47/24 20130101;
A61K 9/1274 20130101 |
International
Class: |
A61K 38/31 20060101
A61K038/31; A61K 47/10 20060101 A61K047/10; A61K 9/00 20060101
A61K009/00; A61K 9/127 20060101 A61K009/127; A61K 47/14 20060101
A61K047/14; A61K 47/24 20060101 A61K047/24 |
Claims
1. A method of treating acromegaly, comprising administering to a
patient in need thereof a lipid composition comprising 20 mg of
octreotide or a halide thereof or a physiologically acceptable acid
thereof, wherein the lipid composition is administered to the
patient once every 28 days.+-.7 days or 30 days.+-.7 days.
2. The method of claim 1, wherein the lipid composition is
administered to the patient once every month.
3. The method of claim 1, wherein the lipid composition is
administered as a unit dose.
4. The method of claim 1, wherein the octreotide or the halide
thereof or the physiologically acceptable acid thereof is
octreotide chloride.
5. The method of claim 1, wherein the octreotide or the halide
thereof or the physiologically acceptable acid thereof is the sole
active agent in the lipid composition.
6. The method of claim 1, wherein the lipid composition further
comprises glycerol dioleate, phosphatidylcholine, and ethanol.
7. The method of claim 6, wherein the lipid composition further
comprises propylene glycol.
8. The method of claim 1, wherein the lipid composition consists of
a low viscosity mixture of: a) a lipid component consisting of: at
least 95 wt. % of: a1) 20-80 wt. % of at least one diacyl glycerol
and/or a tocopherol; and a2) 20-80 wt. % of at least one
phosphatidyl choline (PC); and 0-5 wt. % of at least one impurity
associated with components a1) and/or a2); b) 5-16 wt. % of
ethanol; c) 1-12 wt. % of propylene glycol (PG); d) the octreotide
or the halide thereof or the physiologically acceptable acid
thereof; and e) optionally 0.0005-0.2 wt. % at least one
antioxidant selected from the group consisting of ascorbic acid,
ethylenediaminetetraacetic acid (EDTA), and citric acid; wherein
the lipid composition has a viscosity of 10-750 mPas at 20.degree.
C.; wherein the ratio of components a1:a2 is in the range 40:60 to
54:46; wherein the ratio of components b:c is in the range 30:70 to
60:40; wherein the lipid composition forms, or is capable of
forming, at least one liquid crystalline phase structure upon
contact with excess aqueous fluid; and wherein the lipid
composition is for subcutaneous injection.
9. The method of claim 8, wherein component a1) comprises glycerol
dioleate (GDO).
10. The method of claim 8, wherein component a1) is present at a
level of 30-40% by weight.
11. The method of claim 8, wherein component a2) comprises soy
PC.
12. The method of claim 8, wherein component a2) is present at a
level of 30-40% by weight.
13. The method of claim 8, wherein the ratio of components a1:a2 is
in the range 45:55 to 54:46.
14. The method of claim 8, wherein component b) is present at a
level of 6-14 wt. %.
15. The method of claim 8, wherein component c) is present at a
level of 6-12 wt. %.
16. The method of claim 8, wherein component d) is octreotide,
octreotide chloride, or octreotide acetate.
17. The method of claim 8, wherein component d) is present at a
level of 0.1 to 10 wt. %.
18. The method of claim 8, wherein the value C.sub.max/C.sub.ave of
component d) is reduced by a factor of at least 1.2 relative to
administration of a corresponding lipid composition in which
component c) is absent.
19. The method of claim 8, wherein component e) is present.
20. The method of claim 8, wherein the ratio of components c:e is
in the range 1:50 to 1:1500.
21. The method of claim 8, wherein component a1) consists of
glycerol dioleate (GDO), component a2) consists of PC, component c)
consists of PG, component d) consists of octreotide chloride, and
component e) is present.
22. The method of claim 8, wherein components b) and c) are present
in approximately equal amounts.
23. The method of claim 22, wherein the ratio of components b:c is
50:50.
24. The method of claim 1, comprising administering the lipid
composition by a pre-filled administration device.
25. The method of claim 8, wherein the pre-filled administration
device is a syringe, a pre-filled syringe, or a needle-less
injector.
26. The method of claim 1, administering the lipid composition by
subcutaneous injection.
27. The method of claim 1, comprising administering the lipid
composition in a volume of about 1 mL.
28. The method of claim 1, wherein the patient has a maximum blood
plasma concentration (C.sub.max)/average blood plasma concentration
(C.sub.ave) of octreotide (at steady state) ratio of less than 50,
for each once every month administration.
29. The method of claim 1, wherein the patient has a maximum blood
plasma concentration (C.sub.max)/average blood plasma concentration
(C.sub.ave) of octreotide (at steady state) ratio of less than 15,
for each once every month administration.
30. The method of claim 1, wherein the patient has an average blood
plasma concentration (C.sub.ave)/minimum blood plasma concentration
(C.sub.min) of octreotide (at steady state) ratio of less than 50,
for each once every month administration.
31. The method of claim 1, wherein the patient has an average blood
plasma concentration (C.sub.ave)/minimum blood plasma concentration
(C.sub.min) of octreotide (at steady state) ratio of less than 15,
for each once every month administration.
32. The method of claim 1, wherein the patient has an area under a
plasma concentration against time curve during the first 24 hours
of the one-month dosing of less than 20% of the area under the
curve for the entire curve (measured or extrapolated from time 0 to
infinity or from time 0 to the last sampling time point), for each
once every month administration.
33. The method of claim 1, wherein the patient has an area under a
plasma concentration against time curve during the first 24 hours
of the one-month dosing of less than 15% of the area under the
curve for the entire curve (measured or extrapolated from time 0 to
infinity or from time 0 to the last sampling time point), for each
once every month administration.
34. The method of claim 1, wherein the patient has an area under a
plasma concentration against time curve during the first 24 hours
of the one-month dosing of less than 10% of the area under the
curve for the entire curve (measured or extrapolated from time 0 to
infinity or from time 0 to the last sampling time point), for each
once every month administration.
35. The method of claim 1, wherein the patient self-administers the
lipid composition.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional application of U.S.
application Ser. No. 14/117,994, filed on Feb. 20, 2014, which is a
.sctn. 371 application of International Application No.
PCT/EP2012/059917, filed May 25, 2012, which claims priority to
U.S. Application No. 61/489,886, filed on May 25, 2011, each of
which is incorporated herein by reference.
SEQUENCE LISTING
[0002] The Sequence Listing submitted herewith is an ASCII text
file (2021-06-17_Sequence_Listing.text, created on Jun. 16, 2021,
559 bytes) via EFS-Web is hereby incorporated by reference.
FIELD OF THE INVENTION
[0003] The present invention relates to formulation precursors
(pre-formulations) for the in situ generation of compositions for
the controlled release of peptide active agents, and methods of
treatment with such formulations. In particular, the invention
relates to high-loading pre-formulations of amphiphilic components
and at least one peptide active agent for parenteral application,
which undergo phase transition upon exposure to aqueous fluids,
such as body fluids, thereby forming a controlled release
composition.
BACKGROUND TO THE INVENTION
[0004] Many bioactive agents including pharmaceuticals, nutrients,
vitamins and so forth have a "functional window". That is to say
that there is a range of concentrations over which these agents can
be observed to provide some biological effect. Where the
concentration in the appropriate part of the body (e.g. locally or
as demonstrated by serum concentration) falls below a certain
level, no beneficial effect can be attributed to the agent.
Similarly, there is generally an upper concentration level above
which no further benefit is derived by increasing the
concentration. In some cases increasing the concentration above a
particular level results in undesirable or even dangerous
effects.
[0005] Some bioactive agents have a long biological half-life
and/or a wide functional window and thus may be administered
occasionally, maintaining a functional biological concentration
over a substantial period of time (e.g. 6 hours to several days).
In other cases the rate of clearance is high and/or the functional
window is narrow and thus to maintain a biological concentration
within this window regular (or even continuous) doses of a small
amount are required. This can be particularly difficult where
non-oral routes of administration (e.g. parenteral administration)
are desirable or necessary, since self-administration may be
difficult and thus cause inconvenience and/or poor compliance. In
such cases it would be advantageous for a single administration to
provide active agent at a therapeutic level over the whole period
during which activity is needed.
[0006] There is an enormous potential in the use of peptides
(including proteins) for treating various disease states, as well
as in prophylaxis and in improving general health and well-being of
subjects. However, the performance of administered peptide agents
is generally limited due to poor bioavailability, which in turn is
caused by the rapid degradation of peptides and proteins in
biological fluids. This increases the dose which must be
administered and in many cases restricts the effective routes of
administration. These effects are further exaggerated by the often
limited permeability of peptides and proteins across biological
membranes.
[0007] Peptides and proteins that are administered to the mammalian
body (e.g. orally, intramuscularly etc.) are subject to degradation
by various proteolytic enzymes and systems present throughout the
body. Well known sites of peptidase activity include the stomach
(e.g. pepsin), and the intestinal tract (e.g. trypsin,
chymotrypsin, and others) but other peptidases (e.g.
aminopeptidases, carboxypeptidases, etc.) are found throughout the
body. Upon oral administration, gastric and intestinal degradation
reduces the amount of peptide or protein which potentially could be
absorbed through the intestinal surface lining and thereby
decreases their bioavailability. Similarly, free peptides and
proteins in the mammalian blood stream are also subject to
enzymatic degradation (e.g. by plasma proteases etc.).
[0008] Some patients undergoing treatment will typically require a
therapeutic dose to be maintained for a considerable period and/or
ongoing treatment for many months or years. Thus a depot system
allowing loading and controlled release of a larger dose over a
longer period would offer a considerable advantage over
conventional delivery systems.
[0009] Peptides may be delivered by systems such as the Alkermes
Medisorb.RTM. delivery system consisting of microspheres of
biodegradable polymers. Such polymer microsphere formulations must
generally be administered by means of a sizable needle, typically
of 20-gauge or wider. This is necessary as a result of the nature
of the polymeric dosing systems used, which are typically polymer
suspensions.
[0010] Evidently, it would be an advantage to provide a system of
low viscosity, such as a homogeneous solution, dispersion of fine
particles, or L.sub.2 phase, which could be administered easily
through a narrow needle, thus decreasing the discomfort of the
patient during the procedure. This ease of administration is
particularly significant where patients will be on a
self-administration regime and may already be self-administering
several times each day. Providing a sustained formulation with a
duration of a few days, but which is sufficiently complex to
administer that it requires treatment by a healthcare professional
will not be an advantage to all patients over twice-daily or daily
self-administration, and is likely to be more costly. Providing a
formulation which gives sufficiently long duration to justify a
visit to a health professional for administration and/or a
preparation which can be self-administered, and reducing
preparation time of health-care professionals or patients prior to
the actual administration are all important issues.
[0011] The poly-lactate, poly-glycolate and
poly-lactate-co-glycolate polymers typically used for degrading
slow-release formulations are also the cause of some irritation in
at least some patients. In particular, these polymers typically
contain a certain proportion of acetic acid impurity, which will
irritate the injection site on administration. When the polymer
then breaks down, lactic acid and glycolic acid are the degradation
products so that further irritation is caused. As a result of the
combined effects of wide-needle administration and irritant
contents, discomfort at the site of administration and the
formation of connective scar tissue are greater than desirable.
[0012] From a drug delivery point of view, polymer depot
compositions generally have the disadvantage of accepting only
relatively low drug loads and having a "burst/lag" release profile.
The nature of the polymeric matrix, especially when applied as a
solution or pre-polymer, causes an initial burst of drug release
when the composition is first administered. This is followed by a
period of low release, while the degradation of the matrix begins,
followed finally by an increase in the release rate to the desired
sustained profile. This burst/lag release profile can cause the in
vivo concentration of active agent to burst above the functional
window immediately following administration, and then drop back
through the bottom of the functional window during the lag period
before reaching a sustained functional concentration for a period
of time. Evidently, from a functional and toxicological point of
view this burst/lag release profile is undesirable and could be
dangerous. It may also limit the equilibrium concentration which
can be provided due to the danger of adverse effects at the "peak"
point. The presence of a lag phase may furthermore require
supplementary dosing with repeat injections during the start-up
period of depot treatment in order to maintain a therapeutic dose
while the concentrations of active provided from the depot are
sub-functional. For certain polypeptides in particular, it would be
advantageous to minimise the immediate "burst" effect upon
administration of a composition in order to avoid side effects such
as hypoglycaemia.
[0013] One class of peptide hormones which benefits particularly
from a very "low burst", stable in vivo concentration are
Somatostatin analogues. In vivo testing suggests that these
peptides are particularly beneficial when maintained at a steady
plasma concentration. This not only suggests that a depot
composition would be an advantage to avoid "spikes" in
concentration upon administration and/or repeated daily dosing, but
furthermore that such a depot composition should have as flat a
release profile as possible during the therapeutic period.
[0014] Controlled-release formulations are typically generated from
bio-compatible polymers in the form of, for example, implants or
injectable beads. Polymer microsphere formulations must generally
be administered by means of a sizable needle, typically of 20-gauge
or wider. This is necessary as a result of the nature of the
polymeric dosing systems used, which are typically polymer
suspensions. It would be an advantage to provide a system of low
viscosity, such as a homogeneous solution, dispersion of fine
particles, or L.sub.2 phase, which could be administered easily
through a narrow needle, thus decreasing the discomfort of the
patient during the procedure. In the case of diabetic patients,
whether for daytime or nightly use, this ease of administration is
particularly significant because most patients will be frequently
self-administering. Providing a sustained formulation which can
prevent or reduce the risk of hypoglycemia (especially nocturnal
hypoglycemia), but which is sufficiently complex to administer that
it requires treatment by a healthcare professional is unlikely to
be successful, because the lifestyle disruption involved with such
complex administrations, as well as the costs involved would be too
great. Providing a formulation which can be self-administered, and
which is sufficiently straightforward and painless to administer
that patient compliance is not adversely affected is greatly needed
for such situations.
[0015] The manufacture of PLGA microbeads and suspensions is
additionally a considerable difficulty with certain existing depot
systems. In particular, since the beads are particulate, and
polymers clog membranes, they cannot generally be sterile-filtered
and furthermore, since the PLGA copolymer melts at around
40.degree. C., they cannot be heat-treated for sterility. As a
result, a complex manufacturing process must all be conducted under
conditions of high sterility.
[0016] Further issues with biodegradable polymer microspheres
include complex reconstitution prior to injection and limited
storage stability, due both to aggregation and degradation of the
delivery system and/or active.
[0017] A lipid-based, slow-release composition is described in
WO2006/131730 for GLP-1 and analogues thereof. This is a highly
effective formulation, but the concentration of active agent which
can be included in the formulation is limited by its solubility.
Evidently, a higher concentration of active agent allows for the
possibility of longer duration depot products, products maintaining
a higher systemic concentration, and products having a smaller
injection volume, all of which factors are of considerable
advantage under appropriate circumstances. It would thus be of
considerable value to establish a way by which higher
concentrations of active agents could be included in a lipid-based
depot formulation.
[0018] The present inventors have now established that by providing
a pre-formulation comprising at least one neutral diacyl glycerol
and/or a tocopherol, at least one phosphatidyl choline, at least
one biocompatible organic mono-alcoholic solvent, at least one
polar solvent, at least one peptide active agent and optionally at
least one antioxidant in a low viscosity phase, such as molecular
solution or L.sub.2 (reversed micellar) phase, a pre-formulation
may be generated addressing many of the shortfalls of known depot
formulations, and which may be applied to provide a controlled
release of peptide active agent. By use of specific components in
carefully selected ratios, and in particular with a mixture of an
alcohol and a polar solvent, a depot formulation can be generated
having a combination of properties exceeding the performance of
even the known lipid controlled-release compositions.
[0019] In particular, the pre-formulation shows a highly
advantageous release profile, is easy to manufacture, may be
sterile-filtered, has low viscosity (allowing easy and less painful
administration typically through a narrow needle), allows a high
level of bioactive agent to be incorporated (thus potentially
allowing a smaller amount of composition and/or active agent to be
used), requires shallow injection and/or forms a desired
non-lamellar depot composition in vivo having a "non-burst" release
profile. The compositions are also formed from materials that are
non-toxic, biotolerable and biodegradable, which can be
administered by i.m., or s.c. and are suitable for
self-administration. The pre-formulation may additionally have a
very low level of irritation on injection and in preferred cases
causes no irritation at the injection site (including transient
irritation).
[0020] Certain of the formulations of the present invention
generate a non-lamellar liquid crystalline phase following
administration. The use of non-lamellar phase structures (such as
liquid crystalline phases) in the delivery of bioactive agents is
now relatively well established. A most effective lipid depot
system is described in WO2005/117830, and a highly preferred lipid
depot is described in that document. However, there remains scope
for achieving depot formulations having improved performance in
several respects.
[0021] Advantages of the compositions of the present invention over
polymer formulations, such as PLGA spheres, include the ease of
manufacture (including sterilization), handling and use properties
combined with low initial release ("non-burst profile") of active
agent. This may be defined such that the area under a plasma
concentration against time the curve during the first 24 hours of a
one-month dosing period is less than 20% of the area under the
curve for the entire curve (measured or extrapolated from time 0 to
infinity or from time 0 to the last sampling time point), more
preferably less than 15% and most preferable less than 10%. This
applies particularly to the acyl saccharide and lipid aspects of
the invention and is discussed in more detail in WO 2005/117830.
Furthermore, it may be defined such that the maximum plasma
concentration of active agent in vivo following injection of the
pre-formulation (Cmax) is no more than 10 times, preferably no more
than 8 times and most preferably no more than 5 times the average
plasma concentration during the therapeutic period (Cave).
SUMMARY OF THE INVENTION
[0022] The present invention provides a pharmaceutical formulation
comprising an appropriate combination of lipid excipients, organic
alcoholic solvent, polar solvent, peptide active agent and certain
optional components, that can be used as a depot-precursor
formulation (referred to herein for brevity as a pre-formulation)
to address one or more of the needs described above.
[0023] In a first aspect, the invention therefore provides a
pre-formulation comprising a low viscosity mixture of:
TABLE-US-00002 a. 20-80 wt. % of at least one diacyl glycerol
and/or a tocopherol; b. 20-80 wt. % of at least one phosphatidyl
choline (PC); c. 5-20 wt. % of at least one biocompatible, organic
mono-alcoholic solvent; d. up to 20 wt. % polar solvent e. at least
one peptide active agent; f. optionally at least one
antioxidant;
wherein the ratio of components a:b is in the range 40:60 to 54:46;
wherein the pre-formulation forms, or is capable of forming, at
least one liquid crystalline phase structure upon contact with
excess aqueous fluid.
[0024] Such compositions will preferably comprise GDO, ethanol,
water/propylene glycol and/or EDTA as components a), c), d) and f)
respectively. Component e) is preferably at least one somatostatin
analogue, as described herein.
[0025] In a second embodiment, the invention correspondingly
provides a process for the formation of a pre-formulation suitable
for the administration of a peptide bioactive agent to a
(preferably mammalian) subject, said process comprising forming a
low viscosity mixture of:
TABLE-US-00003 a) 20-80 wt. % of at least one diacyl glycerol
and/or a tocopherol; b) 20-80 wt. % of at least one phosphatidyl
choline (PC); c) 5-20 wt. % of at least one biocompatible, organic
mono-alcoholic solvent; d) up to 20 wt. % polar solvent e) at least
one peptide active agent; f) optionally at least one
antioxidant;
wherein the ratio of components a:b is in the range 40:60 to 54:46;
and dissolving or dispersing at least one peptide active agent
(preferably a somatostatin analogue) in the low viscosity mixture,
or in at least one of components a), b), c), d) and optionally f)
prior to forming the low viscosity mixture. Such a pre-formulation
will typically be one as described herein.
[0026] The preformulations are highly useful for the controlled and
sustained release of peptide active, especially those requiring or
benefiting from a very flat release profile and/or minimal "burst"
upon administration. In a corresponding embodiment, the invention
therefore provides for the use of a low viscosity mixture of:
TABLE-US-00004 a) 20-80 wt. % of at least one diacyl glycerol
and/or a tocopherol; b) 20-80 wt. % of at least one phosphatidyl
choline (PC); c) 5-20 wt. % of at least one biocompatible, organic
mono-alcoholic solvent; d) up to 20 wt. % polar solvent e) at least
one peptide active agent; f) optionally at least one
antioxidant;
wherein the ratio of components a:b is in the range 40:60 to 54:46;
in the manufacture of a pre-formulation for use in the sustained
administration of said peptide active agent. Such a low viscosity
mixture will preferably be one described herein.
[0027] The peptide active agents in the formulations of the present
invention are preferably pharmaceutically active. That is to say
that they provide a therapeutic, palliative and/or prophylactic
effect when administered to a suitable subject (typically being one
in need of such an effect). IN a further embodiment, the invention
therefore provides a method for the treatment of a human or
non-human mammalian subject comprising administering to said
subject a pre-formulation as described herein.
[0028] Such a method may be for the treatment of a human or
non-human mammalian subject in need thereof to combat, (e.g. cure,
improve, prevent or ameliorate the symptoms of) at least one
condition selected from acromegaly, cancers, carcinomas, melanomas,
tumours expressing at least one somatostatin receptor,
sst(2)-positive tumours, sst(5)-positive tumours, prostate cancers,
gastro-entero-pancreatic neuroendocrine (GEP NE) tumours, carcinoid
tumours, insulinomas, gastrinomas, vasoactive intestinal peptide
(VIP) tumours and glucagonomas, elevated growth hormone (GH),
elevated insulin-like growth factor I (IGF-I), varicial bleeding
(especially espohageal), chemotherapy induced gastro intestinal
problems (such as diarrhea), lymphorrhea, diabetic retinopathy,
thyroid eye disease, obesity, pancreatitis, and related conditions.
Such methods are particularly applicable where component e) is at
least one somatostatin analogue, as described herein. The
preformulations as described herein for use in such methods form a
further aspect of the invention.
[0029] Correspondingly, in a further aspect, the present invention
provides the use of a low viscosity mixture of:
TABLE-US-00005 a) 20-80 wt. % of at least one diacyl glycerol
and/or a tocopherol; b) 20-80 wt. % of at least one phosphatidyl
choline (PC); c) 5-20 wt. % of at least one biocompatible, organic
mono-alcoholic solvent; d) up to 20 wt. % polar solvent e) at least
one peptide active agent; f) optionally at least one
antioxidant;
wherein the ratio of components a:b is in the range 40:60 to 54:46;
in the manufacture of a low viscosity pre-formulation medicament
for use in the in vivo formation of a depot for treatment of at
least one condition selected from acromegaly, cancers, carcinomas,
melanomas, tumours expressing at least one somatostatin receptor,
sst(2)-positive tumours, sst(5)-positive tumours, prostate cancers,
gastro-entero-pancreatic neuroendocrine (GEP NE) tumours, carcinoid
tumours, insulinomas, gastrinomas, vasoactive intestinal peptide
(VIP) tumours and glucagonomas, elevated growth hormone (GH),
elevated insulin-like growth factor I (IGF-I), varicial bleeding
(especially espohageal), chemotherapy induced gastro intestinal
problems (such as diarrhea), lymphorrhea, diabetic retinopathy,
thyroid eye disease, obesity, pancreatitis, and related conditions.
Such uses are particularly applicable where component e) is at
least one somatostatin analogue, as described herein.
[0030] Certain peptide active agents have benefits which are
cosmetic rather than (or in addition to) therapeutic in nature.
Such effects include weight-loss and/or hunger suppression as well
as control over skin or hair pigmentation, hair growth etc. The
present invention therefore additionally provides a method of
cosmetic treatment of a human or non-human mammalian subject
comprising administering to said subject a pre-formulation as
described herein. Such a cosmetic method will generally not be a
method of therapy (i.e. will not have therapeutic or medical
benefit).
[0031] One of the advantages of the formulations of the present
invention over many other controlled-release compositions is that
they are stable to storage in their final form and thus little or
no preparation is required at the time of administration. This
allows the pre-formulations to be ready-to-administer and also to
be supplied in convenient, ready-to-administer form. In a further
aspect, the invention therefore provides a pre-filled
administration device containing a pre-formulation as described
herein. Such a device will generally provide either a single
administration or multiple administrations of a composition which
will deliver, for example, a dosage of active agent in the range of
1 .mu.g to 5 mg/day.
[0032] In a further aspect the invention provides a kit comprising
said administration device according to the invention.
[0033] The kit can optionally contain instructions for subcutaneous
or intramuscular administration of said composition. All
compositions described herein are suitable for use in such a kit
and may thus be contained therein.
[0034] The kits of the invention can optionally include additional
administration components such as needles, swabs, and the like and
will optionally contain instructions for administration.
BRIEF SUMMARY OF THE ATTACHED FIGURES
[0035] FIG. 1a. IVR profile of formulations 911 to 918
[0036] FIG. 1b. IVR profile of formulations 1006, 1007, and
1010.
[0037] FIG. 2: Peptide Content and Purity (expressed as % of the
corresponding values obtained for the reference samples stored at
<-15.degree. C.) after storage of formulations G and H for 7
days at 70.degree. C.
[0038] FIG. 3. PK-11-413, dose normalized
[0039] FIG. 4: PK-11-425, leuprolide plasma concentration versus
time over 21 days for formulations 49 and 50.
DETAILED DESCRIPTION OF THE INVENTION
[0040] The formulations of the present invention generate a
non-lamellar liquid crystalline phase following administration. The
use of non-lamellar phase structures (such as liquid crystalline
phases) in the delivery of bioactive agents is now relatively well
established. A most effective lipid depot system is described in
WO2005/117830, and a suitable lipid matrix for use in the present
invention is described in that document, the full disclosure of
which is hereby incorporated herein by reference. For a description
of the most favourable phase structures of such formulations,
attention is drawn to the discussion in WO2005/117830 and
particularly to page 29 thereof.
[0041] All % are specified by weight herein throughout, unless
otherwise indicated. Furthermore, the % by weight indicated is the
% of the total pre-formulation including all of the components
indicated herein. The pre-formulations can optionally consist of
essentially only the components indicated herein (including where
appropriate additional optional components indicated herein below
and in the attached claims) and in one aspect consist entirely of
such components.
[0042] The lipid-based systems described herein comprise lipid
components a) and b), plus organic mono-alcoholic solvent (c),
polar solvent (d), peptide active agent (e) and optional
antioxidant (f) components.
[0043] Preferably the pre-formulation according to the invention
has an L.sub.2 phase structure. Preferably the pre-formulation
forms a non-lamellar (e.g. liquid crystalline) phase following
administration.
[0044] The present inventors have now surprisingly established that
by appropriate choice of types, absolute amounts and ratios of
lipid components along with a peptide active agent and at least two
solvents including an alcohol and at least one polar solvent, the
release properties of the depot compositions formed from the
pre-formulations of the invention can be rendered highly
advantageous. In particular, by using a mixture of an alcohol and a
polar solvent (especially at the ratios close to 1:1 described
herein), the advantages of the alcohol solvent on the release
profile can be maintained while other properties such as the
comfort on administration and/or the viscosity of the formulation
can be improved. Alternatively or in addition to this, the release
profile of the active agent can be made remarkably level, with the
maximum plasma concentration in vivo being only a small multiple of
the average or even minimum concentration during the dosing period.
Such advantages apply even in comparison with other lipid depot
compositions, which in themselves offer previously unobtainable
standards in controlled release.
[0045] It is important, particularly with certain peptide active
agents, such as somatostatin analogues, to control the peak
concentration (Cmax) of drug in the plasma to a level equal to or
less than that tolerable to the subject, for example to avoid
side-effects such as flushing or severe nausea, while providing or
achieving a therapeutically effective level over the desired period
of release. Generally, the average concentration during the period
of release before the next dose is administered, Cave, falls within
the therapeutic range. Control over the maximal (Cmax) and minimum
(Cmin) concentrations is also important in order to achieve the
desired treatment over time. In one embodiment, the initial burst
is not the Cmax of the release profile.
[0046] Whether or not the initial burst is also the Cmax,
preferably the Cmax/Cave ratio is less than 50, preferably less
than or equal to 15, more preferably less than or equal to 10, even
more preferably less than or equal to 5. Furthermore, it is
preferred that the Cave/Cmin ratio is not more than 50, preferably
less than or equal to 15, more preferably less than or equal to 10,
even more preferably less than or equal to 5. Cmax is defined as is
known in the art, as the peak or maximal plasma concentration
observed during the period of release before the next dose is
administered and Cave is defined as the average plasma
concentration during that period of release. Cmin is
correspondingly the minimal concentration during that period. Cave
can be calculated by calculating the drug present in the plasma as
area under the curve (AUC) over the selected period of time,
generally the entire period of release before the administration of
the next dose, and dividing by that period of time.
Component a)--Diacyl Glycerol
[0047] Preferable ranges for component a) are 20-80 wt. %,
preferably 30-70 wt. %, more preferably 33-60% (e.g. 43-60%),
particularly 38 to 43%. Preferable ranges of component b) are 20-80
wt. %, preferably 30-70 wt. %, more preferably 33-55% (e.g.
35-55%), particularly 38 to 43%.
[0048] Ratios of a:b are typically 40:60 to 70:30, preferably 45:55
to 55:45 and more preferably 40:60 to 54:46. Ratios of around 50:50
(e.g. .+-.2) are highly effective.
[0049] Component "a" as indicated herein is preferably at least one
diacyl glycerol (DAG) and thus has two non-polar "tail" groups. The
two non-polar groups may have the same or a differing number of
carbon atoms and may each independently be saturated or
unsaturated. Examples of non-polar groups include C.sub.6-C.sub.32
alkyl and alkenyl groups, which are typically present as the esters
of long chain carboxylic acids. These are often described by
reference to the number of carbon atoms and the number of
unsaturations in the carbon chain. Thus, CX:Z indicates a
hydrocarbon chain having X carbon atoms and Z unsaturations.
Examples particularly include lauroyl (C12:0), myristoyl (C14:0),
palmitoyl (C16:0), phytanoyl (C16:0), palmitoleoyl (C16:1),
stearoyl (C18:0), oleoyl (C18:1), elaidoyl (C18:1), linoleoyl
(C18:2), linolenoyl (C18:3), arachidonoyl (C20:4), behenoyl (C22:0)
and lignoceroyl (C24:9) groups. Thus, typical non-polar chains are
based on the fatty acids of natural ester lipids, including
caproic, caprylic, capric, lauric, myristic, palmitic, phytanic,
palmitolic, stearic, oleic, elaidic, linoleic, linolenic,
arachidonic, behenic or lignoceric acids, or the corresponding
alcohols. Preferable non-polar chains are palmitic, stearic, oleic
and linoleic acids, particularly oleic acid.
[0050] Mixtures of any number of diacyl lipids may be used as
component a). Preferably this component will include at least a
portion of C18 lipids (e.g. DAG having one or more C18:0, C18:1,
C18:2 or C18:3 non-polar groups), such as glycerol dioleate (GDO)
and/or glycerol dilinoleate (GDL). A highly preferred example is
DAG comprising at least 50%, preferably at least 80% and even
comprising substantially 100% GDO.
[0051] Since GDO and other diacyl glycerols are products derived
from natural sources, there is generally a certain proportion of
"contaminant" lipid having other chain lengths etc. In one aspect,
GDO as used herein is thus used to indicate any commercial grade of
GDO with concomitant impurities (i.e. GDO of commercial purity).
These impurities may be separated and removed by purification but
providing the grade is consistent this is rarely necessary. If
necessary, however, "GDO" may be essentially chemically pure GDO,
such as at least 80% pure, preferably at least 85% pure and more
preferably at least 90% pure GDO.
Component b)--Phosphatidyl Choline
[0052] Component "b" in the preferred lipid matrices of the present
invention is at least one phosphatidyl choline (PC). As with
component a), this component comprises a polar head group and at
least one non-polar tail group. The difference between components
a) and b) lies principally in the polar group. The non-polar
portions may thus suitably be derived from the fatty acids or
corresponding alcohols considered above for component a. As with
component a), the PC will contain two non-polar groups. Again, C18
groups are preferred and may be combined with any other suitable
non-polar group, particularly C16 groups.
[0053] The phosphatidyl choline portion, even more suitably than
any diacyl glycerol portion, may be derived from a natural source.
Suitable sources of phospholipids include egg, heart (e.g. bovine),
brain, liver (e.g. bovine) and plant sources including soybean.
Such sources may provide one or more constituents of component b,
which may comprise any mixture of phospholipids. Any single PC or
mixture of PCs from these or other sources may be used, but
mixtures comprising soy PC or egg PC are highly suitable. The PC
component preferably contains at least 50% soy PC or egg PC, more
preferably at least 75% soy PC or egg PC and most preferably
essentially pure soy PC or egg PC.
[0054] In one embodiment applicable to all aspects of the
invention, component b) comprises PC. Preferably the PC is derived
from soy. Preferably the PC comprises 18:2 fatty acids as the
primary fatty acid component with 16:0 and/or 18:1 as the secondary
fatty acid components. These are preferably present in the PC at a
ratio of between 1.5:1 and 6:1. PC having approximately 60-65%
18:2, 10 to 20% 16:0, 5-15% 18:1, with the balance predominantly
other 16 carbon and 18 carbon fatty acids is preferred and is
typical of soy PC.
[0055] In an alternative but equally preferred embodiment, the PC
component may comprise synthetic dioleoyl PC. This is believed to
provide increased stability and so will be particularly preferable
for compositions needing to be stable to long term storage, and/or
having a long release period in vivo. In this embodiment the PC
component preferably contains at least 50% synthetic dioleoyl PC,
more preferably at least 75% synthetic dioleoyl PC and most
preferably essentially pure synthetic dioleoyl PC. Any remaining PC
is preferably soy or egg PC as above.
[0056] Since the pre-formulations of the invention are to be
administered to a subject for the controlled release of a peptide
active agent, it is important that the components are
biocompatible. In this regard, the preferred lipid matrices for use
in the pre-formulations of the present invention are highly
advantageous since both PC and DAGs are well tolerated and are
broken down in vivo into components that are naturally present in
the mammalian body.
[0057] Synthetic or highly purified PCs, such as dioleoyl
phosphatidy choline (DOPC) are highly appropriate as all or part of
component b). The synthetic dioleoyl PC is most preferably
1,2-dioleoyl-sn-glycero-3-phosphocholine, and other synthetic PC
components include DDPC
(1,2-Didecanoyl-sn-glycero-3-phosphocholine);
DEPC(1,2-Dierucoyl-sn-glycero-3-phosphocholine);
DLOPC(1,2-Dilinoleoyl-sn-glycero-3-phosphocholine);
DLPC(1,2-Dilauroyl-sn-glycero-3-phosphocholine);
DMPC(1,2-Dimyristoyl-sn-glycero-3-phosphocholine);
DOPC(1,2-Dioleoyl-sn-glycero-3-phosphocholine);
DPPC(1,2-Dipalmitoyl-sn-glycero-3-phosphocholine);
DSPC(1,2-Distearoyl-sn-glycero-3-phosphocholine);
MPPC(1-Myristoyl-2-palmitoyl-sn-glycero 3-phosphocholine);
MSPC(1-Myristoyl-2-stearoyl-sn-glycero-3-phosphocholine); PMPC
(1-Palmitoyl-2-myristoyl-sn-glycero-3-phosphocholine);
POPC(1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine); PSPC
(1-Palmitoyl-2-stearoyl-sn-glycero-3-phosphocholine); SMPC
(1-Stearoyl-2-myristoyl-sn-glycero-3-phosphocholine);
SOPC(1-Stearoyl-2-oleoyl-sn-glycero-3-phosphocholine); and SPPC
(1-Stearoyl-2-palmitoyl-sn-glycero-3-phosphocholine), or any
combination thereof.
[0058] In some circumstances, such as the absence of preserving
agents such as EDTA, the use of synthetic or highly purified PCs
(e.g. DOPC) may provide greater stability for the active agent in
the formulations. Thus in one embodiment, component b) may comprise
(e.g. may comprise at least 75%) synthetic or highly purified (e.g.
purity >90%) PCs (e.g. DOPC). This may particularly be in the
absence of chelating agents such as EDTA. In an alternative
embodiment, component b) may comprise (e.g. comprise at least 75%)
naturally derived PCs, such as soy PC or egg PC. This will
particularly be where at least one stabilising component (such as
an antioxidant, chelator etc) is included in the precursor
formulation.
[0059] A particularly favoured combination of components a) and b)
are GDO with PC, especially GDO with soy PC and/or DOPC.
Appropriate amounts of each component suitable for the combination
are those amounts indicated herein for the individual components in
any combination. This applies also to any combinations of
components indicated herein, where context allows.
[0060] The ratio of components a:b is in the range 40:60 to 54:46.
Preferably the a:b ratio is in the range 45:55 to 54:46, more
preferably 47:53 to 53:47. Most preferably the a:b ratio is
approximately 50:50.
Component c)--Organic Mono-Alcoholic Solvent
[0061] Component c) of the pre-formulations of the invention is an
organic mono-alcoholic solvent. Since the pre-formulation is to
generate a depot composition following administration (e.g. in
vivo), typically upon contact with excess aqueous fluid, it is
desirable that this solvent be tolerable to the subject and be
capable of mixing with the aqueous fluid, and/or diffusing or
dissolving out of the pre-formulation into the aqueous fluid.
Solvents having at least moderate water solubility are thus
preferred.
[0062] Most preferably component c) comprises or consists of
ethanol, propanol, ispropanol, or mixtures thereof. Most preferably
component c) comprises or consists of ethanol.
[0063] In a preferred embodiment, the solvent is such that a
relatively small addition to a mixture comprising a) and b) (i.e.
preferably below 15%) gives large viscosity reductions, of one
order of magnitude or more. As described herein, the addition of
10% organic mono-alcohol solvent can give a reduction of two or
more orders of magnitude in viscosity over the solvent-free
composition, or over a depot containing only a polar solvent such
as water, or glycerol.
[0064] The amount of component c) in the pre-formulation will have
a considerable effect upon several features. In particular, the
viscosity and the rate (and duration) of release will alter
significantly with the solvent level. The amount of solvent will
thus be at least sufficient to provide a low viscosity mixture but
will additionally be determined so as to provide the desired
release rate. This may be determined by routine methods in view of
the Examples below. Typically a level of 0.1 to 35%, particularly 5
to 25% solvent will provide suitable release and viscosity
properties. This will preferably be 5 to 16% (e.g. 6 to 14%) and an
amount of around 8% (e.g. 8.+-.2%) is highly effective.
[0065] As indicated above, the amount of component c) in the
pre-formulations of the invention will be at least sufficient to
provide a low viscosity mixture (e.g. a molecular solution, see
above) of components a), b), c) and d) and optionally f) and will
be easily determined for any particular combination of components
by standard methods.
[0066] The phase behaviour may be analysed by techniques such as
visual observation in combination with polarized light microscopy,
X-ray scattering and diffraction techniques, nuclear magnetic
resonance, and cryo-transmission electron microscopy (cryo-TEM) to
look for solutions, L.sub.2 or L.sub.3 phases, or liquid
crystalline phases or as in the case of cryoTEM, dispersed
fragments of such phases. Viscosity may be measured directly by
standard means. As described above, an appropriate practical
viscosity is that which can effectively be syringed and
particularly sterile filtered. This will be assessed easily as
indicated herein. Typical organic mono-alcoholic solvents suitable
for use in the invention include at least one solvent selected from
ethanol, propanol, isopropanol, and benzyl alcohol, particularly
ethanol.
[0067] A highly preferred combination for components a), b) and c)
is soy PC, GDO and ethanol. As indicated above, appropriate amounts
of each component suitable for the combination are those amounts
indicated herein for the individual components, in any
combination.
[0068] It is preferable that little or none of component c)
contains halogen substituted hydrocarbons since these tend to have
lower biocompatibility.
[0069] Component c) as used herein may be a single solvent or a
mixture of suitable solvents but will generally be of low
viscosity. This is important because one of the key aspects of the
present invention is that it provides pre-formulations that are of
low viscosity and a primary role of a suitable solvent is to reduce
this viscosity. This reduction will be a combination of the effect
of the lower viscosity of the solvent and the effect of the
molecular interactions between solvent and lipid composition. One
observation of the present inventors is that the oxygen-containing
solvents of low viscosity described herein have highly advantageous
and unexpected molecular interactions with the lipid parts of the
composition, thereby providing a non-linear reduction in viscosity
with the addition of a small volume of solvent.
[0070] The viscosity of the "low viscosity" solvent component c)
(single solvent or mixture) should typically be no more than 18
mPas at 20.degree. C. This is preferably no more than 15 mPas, more
preferably no more than 10 mPas and most preferably no more than 7
mPas at 20.degree. C.
Component d)--Polar Solvent
[0071] Some of the particular benefits of the compositions of the
present invention come through the unexpected finding that the use
of an alcohol solvent in combination with a polar solvent such as a
diol or water allows a significant improvement in the performance
of certain lipid-based controlled-release compositions. In
particular, the addition of a diol, such as propylene glycol or
water has been observed to reduce the viscosity of a
lipid/alcohol/active agent formulation without adversely affecting
the release profile of the active agent and/or allows the
proportion of alcohol to be increased without adversely affecting
the release profile and/or allows an improvement in the release
profile. By "adversely affecting the release profile" is intended
to indicate that the ratio of Cmax/Cave is increased and/or the
ratio of Cmax/Cmin is increased (for example increased by a factor
of at least 1.2). Similarly an improvement in the release profile
indicates that the ratio of Cmax/Cave and/or Cmax/Cmin is decreased
(e.g. decreased by a factor of at least 1.2.)
[0072] Although it has previously been suggested that lipid
controlled-release compositions should be formulated substantially
in the absence of water, in order to avoid the conversion to
high-viscosity liquid crystalline phases, it has now furthermore
been established that a small and carefully controlled amount of a
polar solvent such as water can provide considerable benefits. In
particular, the inclusion of this polar solvent (preferably
comprising water) allows further improvements in controlling the
initial release of active agent, allows higher stable loading of
some peptide active agents, provides faster depot formation and/or
provides further reduced discomfort upon injection. Any one of
these factors potentially provides a significant improvement in the
context of therapeutic drug delivery, patient health and/or patient
compliance.
[0073] The pre-formulations of the present invention must thus also
contain a polar solvent, component d). A suitable amount will
typically be greater than 1% by weight of the pre-formulation, for
example 1-30 wt. %, particularly 1.2-20 wt. %, especially 2-18 wt.
%. More preferably component d) is present in the range 5-15 wt. %,
especially 6-12 wt. %. Component d) is preferably water, propylene
glycol or mixtures thereof. In one preferred aspect, the
pre-formulations of the invention contain ethanol as component c)
with water and/or propylene glycol as component d).
[0074] In one embodiment the preformulation comprises at least 1.5%
(e.g. at least 4.5%) water as part of component d) (by weight of
the total composition) with the remainder being propylene glycol.
At least 5% water with the balance of component d) being PG is
preferred. Component d) may comprise or consist of water.
[0075] In an alternative embodiment, component d) may comprise or
consist of propylene glycol.
[0076] Preferably the total level of components c) and d) is not
more than 35 wt. %, preferably not more than 30 wt. %, preferably
10-30 wt. %, most preferably 12-25%.
[0077] The ratio of components c) and d) will also have potential
advantages in the compositions of the invention. In particular, by
inclusion of some polar solvent which is miscible with the
mono-alcohol component (especially water), the slight sensation
that may be caused at the injection site from the alcohol content
can be substantially eliminated. Thus, in one embodiment, the ratio
of components c):d) may be in the range 30:70 to 70:30, more
preferably 40:60 to 60:40. In one embodiment, the amount of alcohol
component c) by weight is no greater than the amount of polar
solvent d). Ratios of c):d) ranging from 30:70 to 50:50 are thus
appropriate in such an embodiment. Approximately equal amounts of
components c) and d) are highly appropriate.
[0078] A highly preferred combination for the lipid matrix aspect
is soy PC, GDO, ethanol, and water/propylene glycol or mixtures
thereof. As indicated above, appropriate amounts of each component
suitable for the combination are those amounts indicated herein for
the individual components, in any combination.
Component e)--Peptide Active Agent
[0079] The pre-formulations of the present invention contain one or
more peptide active agents. Suitable peptide active agents are
disclosed and discussed in detail in US WO 2006/075124 and the
disclosures of that document are incorporated herein by reference.
Suitable peptides for use in the necessary peptides may be
naturally occurring or derived from natural peptides, or may be
chemically modified or wholly synthetic peptide molecules. Any
amino acids may be comprised in the peptides including those
described herein, and the peptides may be chemically or
enzymatically modified.
[0080] Typical peptide actives will be in the range of 500 to
100,000 amu in molecular weight and can evidently include protein
active agents. In one embodiment, the polypeptides can have at
least one cationic charge at neutral and/or physiological pH, and
most preferably will require at least one anionic counter-ion at pH
6.5 or above, preferably at pH 7.5 or above. This counter-ion will
be physiologically acceptable, and may thus be a halide or the ion
of a physiologically acceptable acid. Acetate counter ions and/or
chloride ions are particularly preferred and therefore in one
embodiment of the invention, the active agent is a peptide acetate
and/or chloride.
[0081] Examples of suitable classes of peptides include peptide
hormones and synthetic analogues (such as luteinizing-hormone
releasing hormone (LHRH) and analogues (eg, leuprorelin, goserelin,
buserelin, tryptorelin, degarelix), incretins and incretin mimetics
(such as GLP-1 & analogues or glucose-dependent insulinotropic
peptide (GIP)), glucagon, insulin and analogues, interferons,
vasopressins, calcitonins, etc.), cytokines, antibody fragments
(FAbs; scVFs), antimicrobial peptides (g, corticostatins,
defensins, histatins), specific targeting peptides (e.g., as the
examples described in Current Opinion Genetics & Development
10, 71-77 (2006)), venom peptides (e.g., conopeptides), and
immunogenic peptides (e.g., fragments of proteins used for
vaccination purposes).
[0082] In one embodiment, LHRH analogues (also known as GnRH
analogues) form a preferred group of active agents for use in the
present invention. Preferably such peptides will be structurally
related to GnRH I, II and/or III, and/or one or more of the known
analogues, including those listed here.
[0083] Particularly preferred GnRH analogues are constrained
peptides of 6 to 12 alpha-amino acids, of which particular examples
include those indicated above, and particularly leuprolide and
goserelin, of the sequences indicated above.
[0084] In a further embodiment, GLP-1 and its analogues form a
further preferred group of active agents. GLP-1 analogues will be
peptides, especially of around 30 amino acids, e.g. 20 to 45,
especially 25 to 38. Preferably such peptides will be structurally
related to GLP-1 and/or one or more of the known analogues,
including those listed here. By "GLP-1 analogue", as used herein is
indicated any GLP-1 receptor agonist (or less preferably
antagonist), including naturally occurring forms of GLP-1, either
human or from any other species. These analogues are preferably
peptides, peptide derivatives or peptide mimics. Peptide derived
GLP-1 agonists are most preferred, especially GLP-1(7-37),
GLP-1(7-36)amide, Liraglutide (Novo Nordisk), AVE-010
(ZP10--Zealand Pharma--Sanofi-Aventis), TH0318 (TheraTechnologies),
CJC-1131 (ConjuChem), LY548806 (Lilly), Exenatide. (Byetta,
Amylin-Lilly) and their derivatives.
[0085] In the peptide actives of the present invention, peptides
may contain only amino acids selected from those 20 .alpha.-amino
acids indicated in the genetic code, or more preferably may contain
their isomers and other natural and non-natural amino acids,
(generally .alpha., .beta. or .gamma. amino acids) and their
analogues and derivatives.
[0086] Amino acid derivatives are especially useful at the termini
of the peptides, where the terminal amino or carboxylate group may
be substituted by or with any other functional group such as
hydroxy, alkoxy, carboxy (on the N-terminal end), ester, amide,
thio, amido, amino (on the C-terminal end), alkyl amino, di- or
tri-alkyl amino, alkyl (by which is meant, herein throughout
C.sub.1-C.sub.20 alkyl, preferably C.sub.1-C.sub.18 alkyl e.g.
methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-, sec- or t-butyl
etc.), aryl (e.g phenyl, benzyl, napthyl etc), heteroaryl, or other
functional groups, preferably with at least one heteroatom and
preferably having no more than 20 atoms in total, more preferably
no more than 10 and most preferably not more than 6 atoms
(optionally excluding hydrogens).
[0087] In one preferred embodiment of the present invention, the
peptide active agent will comprise a somatostatin, or any analogue
or derivative thereof.
[0088] Somatostatin has two active forms produced by alternative
cleavage of a single preproprotein: one of 14 amino acids, the
other of 28 amino acids. Somatostatin 1-14 is a cyclic peptide
hormone having the sequence
Ala-Gly-Cys-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys (SEQ ID NO:
1), where the two cystine residues are connected by a disulphide
bridge to generate a type II .beta.-turn at the key binding
sequence of Phe-Trp-Lys-Thr (SEQ ID NO: 2). Somatostatin is a
natural peptide hormone also known as Growth Hormone Release
Inhibiting Factor and has a role as an antagonist of insulin,
glucogen and certain other hormones in the release of somatotrophin
(Human Growth Hormone). The biological half-life of natural
Somatostatin is very short (1-3 minutes) and so in itself is
difficult to formulate as a viable therapeutic. However, the lipid
depot compositions of the present invention are highly effective
for short-lived active agents and an increasing number of
somatostatin analogues are becoming available with higher
activities and/or longer clearance times in vivo.
[0089] Somatostatin analogues, such as octreotide, lanreotide,
vapreotide, pasireotide (SOM 230) and related peptides, are used or
indicated in the treatment of a variety of conditions where they
are typically administered over an extended period.
[0090] Octreotide, for example, is the synthetic octa-peptide with
sequence D-Phe-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-ol (2-7 disulphide
bridge) and is typically administered as the acetate salt. Several
clinical studies also feature the octreotide pamoate. This
derivative retains the key Phe-(D)Trp-Lys-Thr .beta.-turn but, in
contrast to the natural hormone, has a terminal half-life of around
1.7 hours. Octreotide is used in treatment of conditions including
carcinoid tumours and acromegaly, and after an initial dose is
typically given over a sustained period of weeks, or more commonly
many months or years. In addition, somatostatin analogues are
indicated in the treatment of many cancers since a wide variety of
tumours are found to express somatostatin receptors. Of particular
interest are those expressing the "sst(2)" and/or "sst(5)"
receptor.
[0091] The most common "simple" formulation of Octreotide is
"Sandostatin".RTM. from Novartis. This is a solution for
subcutaneous (s.c) injection and a 100 .mu.g dose reaches a peak
concentration of 5.2 ng/ml at 0.4 hours post injection. The
duration of action can be up to 12 hours but s.c. dosing is
generally carried out every 8 hours. Evidently, s.c. injection 3
times daily for periods of months or years is not an ideal dosing
regime.
[0092] In order to avoid the need for multiple daily injections of
octreotide, a further formulation is available; "Sandostatin
LAR".RTM., again from Novartis. This is a formulation of octreotide
in poly lactic co-glycolic acid microspheres which, after
resuspension, may be administered by intra muscular (i.m.)
injection.
[0093] Carcinoid tumours are intestinal tumour arising from
specialised cells with paracrine functions (APUD cells). The
primary tumour is commonly in the appendix, where it is clinically
benign. Secondary, metastatic, intestinal carcinoid tumours secrete
excessive amounts of vasoactive substances, including serotonin,
bradykinin, histamine, prostaglandins, and polypeptide hormones.
The clinical result is carcinoid syndrome (a syndrome of episodic
cutaneous flushing, cyanosis, abdominal cramps, and diarrhea in a
patient with valvular heart disease and, less commonly, asthma and
arthropathy). These tumours may grow anywhere in the
gastrointestinal tract (and in the lungs) with approximately 90% in
the appendix. The remainder occurs in the ileum, stomach, colon or
rectum. Currently, treatment of carcinoid syndrome starts with i.v.
bolus injection followed by i.v. infusion. When sufficient effect
on symptoms has been established, treatment with a depot
formulation of octreotide formulated in ploy lactic-co-glycolic
acid (PLGA) microspheres is started. However, during the first two
weeks or more after injection of the depot, daily s.c. injections
with octreotide are recommended to compensate for the slow release
from the PLGA spheres.
[0094] Acromegaly is a rare chronic and insidious hormonal disorder
that occurs when the pituitary gland produces excess growth hormone
(GH). It most commonly affects middle-aged adults and may lead to
premature death.
[0095] Diabetes mellitus, hypertension, and increased risk of
cardiovascular disease are the most serious health consequences of
acromegaly. In addition, patients with acromegaly are at an
increased risk of developing colon polyps, which can become
cancerous. The prevalence of acromegaly is approximately 60 cases
per million population, and the incidence is 3.3 new cases per
million per year. The word acromegaly comes from the Greek words
for "extremities" (acro) and "great" (megaly), because one of the
most common symptoms of this condition is abnormal growth of the
hands and feet.
[0096] Acromegaly is caused by prolonged overproduction of growth
hormone (GH) and excessive production of insulin-like growth
factor-I (IGF-I). In 98 percent of cases, the overproduction of GH
is caused by a pituitary adenoma. The rate of GH production and the
aggressiveness of the tumour vary from patient to patient.
Generally, more aggressive tumours are seen in younger
patients.
[0097] Acromegaly is a severe disease often diagnosed late.
Morbidity and mortality rates are high, in particular, because of
associated cardiovascular, cerebrovascular, and respiratory
disorders and malignancies.
[0098] Treatment of acromegaly is initiated by a period of s.c.
injections three times per day (optimal daily dose=300 .mu.g
octreotide). After the last s.c. dose and providing a suitable
effect is observed then treatment with a depot formulation of
octreotide formulated in poly lactic-co-glycolic acid (PLGA)
microspheres is started. Dose adjustments are made after
measurement of biomarkers (HG and IGF-1), typically after around 3
months.
[0099] The existing octreotide slow release formulation relies upon
a well-established degrading-polymer type of depot formulation.
Typically such formulations are based on a biodegradable polymer
such poly (lactic acid) (PLA) and/or poly (lactic-co-glycolic acid)
(PLGA) and may be in the form of a solution in an organic solvent,
a pre-polymer mixed with an initiator, encapsulated polymer
particles or (as in the case of octreotide) polymer
microspheres.
[0100] In one typical embodiment, the peptide active agent (e.g.
somatostatin analogue) will generally be formulated as 0.02 to 12%
by weight of the total formulation. Typical values will be 0.1 to
10%, preferably 0.2 to 8%, more preferably 0.5 to 6% (e.g. 1 to
3%). These levels may be applied to all aspects of the invention,
where context allows. For octreotide, a further preferred range is
between 0.5 to 4 wt. %, more preferably 1-3 wt. %, and most
preferably 1.5-2.5 wt. %.
[0101] In a related embodiment, the peptide active agent may be
formulated at a level which cannot easily be achieved in the
absence of the polar solvent component of the mixture. In such an
embodiment, the peptide active agent (e.g. Somatostatin analogue)
content is typically at least 0.7%, preferably at least 1%, more
preferably at least 1.8% or at least 2% by weight of formulation.
Levels of at least 3% and at least 4% are achievable with the
present invention, as are loading levels up to 8, 10 or 12%. Such
compositions of the present invention typically not only contain a
very high level of peptide active agent (especially Somatostatin
analogue, e.g. octreotide), as indicated, but are additionally
stable to storage without loss or degradation of the active agent
for considerable periods, as indicated herein. Such periods will
generally be at least a month at 25.degree. C. or at least a month
at 5.degree. C., preferably at least 3 months, and more preferably
at least 6 months at 5.degree. C. or alternatively at 25.degree. C.
These degrees of stability are applicable to all aspects of the
invention, where context allows and relate to stability both of the
active agent and of the phase behaviour of the pre-formulation.
[0102] In a related embodiment, in the situation where a peptide
active agent is highly soluble in the alcohol component, it may be
an advantage to limit this solubility of this agent. Without being
bound by theory, it is thought that excessive solubility in this
alcohol component may result in the alcohol transporting a
significant quantity of active agent out of the depot composition
as it forms in vivo. Therefore, in one embodiment of the present
invention, the polar solvent is used to control the solubility of
the active agent in the preformulation so as aid control of the
release profile.
[0103] In one embodiment, the peptide active agent may be a peptide
which is not a somatostatin analogue (as defined herein). For
example, the peptide active agent may be a peptide which does not
interact as either agonist or antagonist at any of the SST(1) to
SST(5) receptors (especially the corresponding human
receptors).
[0104] In one embodiment, the peptide active agent may be a dual
receptor modulator, having a somatostatin analogue directly
conjugated to a receptor agonist or antagonist for another
receptor. These are referred to herein as "dual receptor agonists.
Dual receptor agonists as indicated herein are peptide compounds
having at least two distinct domains wherein one domain serves as
an agonist for the somatostatin receptor and another serves as an
agonist or antagonist for another biological receptor. Such dual
agonists are distinct from a single non-specific agonist in that,
although the domains may and preferably will be covalently bound
together, the domain serving as somatostatin receptor agonist
resides on a distinct portion of the peptide sequence from the
domain serving to affect the other receptor. That is to say, the
dual agonist is a compound in which a peptide sequence having
somatostatin receptor function and substantially no function at the
second receptor is chemically linked (directly or indirectly) to a
sequence having function at the second receptor and substantially
no somatostatin receptor agonist function.
[0105] In one embodiment, the active agent is not a dual amylin
receptor/GLP-1 receptor agonist compound.
[0106] In a further aspect, the present invention therefore
provides a method for controlling the solubility of a peptide
active agent (such as a somatostatin analogue as described herein)
in a low viscosity mixture comprising:
TABLE-US-00006 a) 20-80 wt. % of at least one diacyl glycerol
and/or a tocopherol; b) 20-80 wt. % of at least one phosphatidyl
choline (PC); c) 5-20 wt. % of at least one biocompatible, organic
mono-alcoholic solvent; e) at least one peptide active agent; f)
optionally at least one antioxidant;
by inclusion of a polar solvent component d) to form a depot
precursor formulation. Use of a polar solvent in such a method
forms a further aspect.
[0107] The pre-formulations and components of the mixture, as well
as their performance etc will evidently correspond to those
described herein for other aspects.
[0108] Similarly, the present invention provides a method for
improving the release profile of a peptide active agent (such as a
somatostatin analogue as described herein) from a depot composition
formed by injection of in a low viscosity mixture comprising:
TABLE-US-00007 a) 20-80 wt. % of at least one diacyl glycerol
and/or a tocopherol; b) 20-80 wt. % of at least one phosphatidyl
choline (PC); c) 5-20 wt. % of at least one biocompatible, organic
mono-alcoholic solvent; e) at least one peptide active agent; f)
optionally at least one antioxidant;
by inclusion of a polar solvent component d) in said low-viscosity
mixture to form a depot precursor formulation. Use of a polar
solvent in such a method forms a further aspect.
[0109] The pre-formulations and components of the mixture, as well
as their performance etc will evidently correspond to those
described herein for other aspects.
[0110] Corresponding methods and uses provide for the reduction of
injection-site discomfort, reduction of viscosity of the
pre-formulation, and/or reduction in initial "burst" release of a
low viscosity mixture comprising:
TABLE-US-00008 a) 20-80 wt. % of at least one diacyl glycerol
and/or a tocopherol; b) 20-80 wt. % of at least one phosphatidyl
choline (PC); c) 5-20 wt. % of at least one biocompatible, organic
mono-alcoholic solvent; e) at least one peptide active agent; f)
optionally at least one antioxidant;
by inclusion of a polar solvent component d) in said low-viscosity
mixture to form a depot precursor formulation. Use of a polar
solvent in such a method forms a further aspect.
[0111] All of the above uses and methods for improving the various
properties of the preformulation and/or the resulting depot
composition are preferably applied without negatively affecting the
release profile of the peptide active agent.
[0112] Where the peptide active agent comprises somatostatin
analogue, (e.g. octreotide), suitable doses for inclusion in the
formulation, and thus the volume of formulation used, will depend
upon the release rate (as controlled, for example by the solvent
type and amount used, the antioxidant content and so forth) and
release duration, as well as the desired therapeutic level, the
activity of the specific agent, and the rate of clearance of the
particular active chosen. Typically an amount of around 0.05 to 40
mg per week of depot duration, preferably 0.1 to 20 mg per week
duration (e.g. 1 to 10 mg per week) for a duration of 1 to 24
weeks, preferably 2 to 16 (e.g. 3, 4, 8, 10 or 12) weeks. In an
alternative embodiment the preformulation may be formulated for
dosing weekly (e.g. every 7.+-.1 days). A total dose of 0.05 to 250
mg per dose would be suitable for providing a therapeutic level for
between 7 and 168 days. This will preferably be 0.1 to 192 mg, e.g.
0.2 to 160 mg, 0.1 to 1.6 mg, 20 to 160 mg etc. Evidently, the
stability of the active and linearity of the release rate will mean
that the loading to duration may not be a linear relationship. A
depot administered every 30 days might have, for example 0.2 to 20
mg or a 90 day depot have 60 to 120 mg of active agent (e.g.
Somatostatin analogue, e.g. octreotide). Evidently also, the
biological half-life of the specific active will be particularly
important. The half-life of somatostatin, is less than 5 minutes,
and so for sustained release, a relatively large amount (e.g.
towards the higher end of the range) will be needed. For an
analogue such as octreotide, with a much longer half-life, the
amount needed will evidently be lower. Appropriate levels for other
actives will be established easily by those of skill in the art by
reference to the known therapeutic level, the desired duration of
action and the volume which is to be injected. A good base
calculation would be to multiply a typical daily dose of the active
agent by the number of day's duration of the depot. The formulation
can then be tested for linearity of release and adjusted as
appropriate.
[0113] It is a remarkable development of the present formulations
that very short half-life peptide active agents, comprising, e.g.
somatostatin and its analogues can be prepared and administered in
a depot precursor of the present invention, and will provide
controlled release over several days or even weeks. This is in
spite of the remarkably short biological half-life of the active
agent (e.g. less than 1 hour, preferably less than 15 minutes, e.g.
less than 5 minutes). Thus, in one embodiment, the active agent has
a half-life of less than 1 hour, e.g. less than 15 minutes and the
preformulation forms a depot which provides sustained release for
at least 7 days, preferably at least 14 days, more preferably at
least 28 days.
[0114] Like essentially all organic molecules, lipids and
biologically active agents are thermodynamically unstable to
oxidation. As a result, many lipid formulations, including those
comprising bioactive agents such as APIs are susceptible to
degradation upon storage, especially by oxidation.
[0115] In a highly preferred embodiment, the lipid matrix aspect is
soy PC, GDO, ethanol, and water/propylene glycol or mixtures
thereof, and the peptide active agent comprises somatostatin or a
somatostatin analogue. As indicated above, appropriate amounts of
each component suitable for the combination are those amounts
indicated herein for the individual components, in any
combination.
Optional Component f)--Antioxidant
[0116] Component f) is an antioxidant. Most preferably it is
selected from ascorbic acid, ethylenediaminetetraacetic acid (EDTA)
and citric acid.
[0117] In all aspects of the invention, component f) is typically
present at a weight ratio of antioxidant to peptide active agent of
1:50 to 1:1500, preferably 1:100 to 1:1300, and most preferably
1:150 to 1:1250. Since typical antioxidants are of lower molecular
weight that the peptide active agents, the proportion by weight of
antioxidant may be relatively small. For example, with a small
molecular weight pH adjuster (e.g. less than 500 amu), 0.0001 to
0.5% of the composition may be antioxidant, preferably 0.0005 to
0.2%, more preferably 0.0008 to 0.1%, e.g. 0.001 to 0.015%.
[0118] Unfortunately, many common antioxidants are not highly
compatible with lipid systems. Indeed, the present inventors have
previously established that some antioxidants commonly used in
previous systems can cause increased degradation of active agents
in a lipid system. This applies particularly to peptide active
agents. The present inventors have therefore analysed a variety of
potential antioxidant compounds and classes for use with lipid
based matrix systems and have surprisingly found that one
particular class of antioxidants is unusually well suited for use
in these systems.
[0119] The antioxidant component is generally included in the range
0.0001 to 0.5% by weight of the total pre-formulation. Around
0.0005 to 0.015% of antioxidant (particularly EDTA) is particularly
preferred, especially in combination with the other preferred
components and ranges indicated herein above and below.
[0120] Stability data using a number of different antioxidants
demonstrate that EDTA antioxidants are surprisingly more efficient
than other antioxidants in suppressing the oxidative degradation of
bioactive agents. EDTA as antioxidant can also show a synergistic
effect in combination with the antioxidants of the present
invention, in maintaining the chemical and physical stability of
the peptide active agent and complete pre-formulation. EDTA has a
stabilising effect on the active agent.
[0121] By "stabilising" is indicated an increase in solubility or
dispensability of a component (especially an active agent) in the
depot delivery system, or alternatively an increase in the
stability of the composition, especially with regard to the
physical and chemical stability of the dissolved or dispersed
active agent. An increase in stability may thus be demonstrated by
dissolution, dispersion or suspension of a greater amount of active
agent in the presence of the antioxidant than would be achieved by
equilibration, such as by agitation for a prolonged period (e.g. 5
days at 25.degree. C.), in the absence of antioxidant. Equally, an
increase in stability may be demonstrated by the chemical and/or
physical stability of a peptide active agent in a lipid formulation
for a greater period than would be observed in the absence of an
antioxidant. This would preferably be tested under conditions of
typical storage, such as 0-5.degree. C., 25.degree. C. and/or
ambient temperature. This is further described herein below.
Optional Additional Components
[0122] In one particularly preferred embodiment of the present
invention, the compositions (preformulations and resulting depots)
do not include fragmentation agents, such as polyethyleneoxide or
poly(ethylene glycol) (PEG) fragmentation agent, e.g. a PEG grafted
lipid and/or surfactant.
[0123] For example, the copmositions preferably do not include
fragmentation agents such as Polysorbate 80 (P80), or other
Polysorbates (e.g. Polysorbate 20), PEGylated phospholipids
(PEG-lipids such as DSPE-PEG(2000), DSPE-PEG(5000), DOPE-PEG(2000)
and DOPE-PEG(5000)), Solutol HS 15, PEGylated fatty acids (e.g.
PEG-oleate), block co-polymers such as Pluronic.RTM. F127 and
Pluronic.RTM. F68, ethoxylated castor oil derivatives (e.g.
Chremophores), PEGylated glyceryl fatty acid esters (such as
TMGO-15 from Nikko Chemicals) and PEGylated tocopherols (such as
d-alpha tocopheryl poly(ethylene glycol) 1000 succinate known as
Vitamin E TPGS from Eastman.
[0124] However, the polypeptide active as a powder (e.g. in the kit
of the invention), as well as active agent dissolved in the lipid
formulation, may gain stability (both storage and in vivo
stability) by certain stabilising additives. Such additives include
sugars (e.g. sucrose, trehalose, lactose etc.), polymers (e.g.
polyols such as carboxy methyl cellulose), amino acids (such as
methionine, glutamate, lysine etc.), lipid-soluble acid components
such as HCl, anionic lipids and/or surface active agents (such as
dioleoyl phosphatidyl glycerol (DOPG), palmitoyloleoyl
phosphatidylglycerol (POPG) and oleic acid (OA)).
[0125] Single-dose formats must remain stable and potent in storage
prior to use, but are disposable after the single use. In one
embodiment, a single dose format is stable at refrigerated
conditions (e.g. 0-5.degree. C.) for at least 12 months.
Furthermore such a preformulation may be stable at room temperature
(e.g. 25.degree. C.) for at least 12 months. Multi-dose formats
must not only remain stable and potent in storage prior to use, but
must also remain stable, potent and relatively free of bacteria
over the multiple-dose use regimen administration period after the
first use in which a seal has been compromised. For this reason
multi-dose formats often require a anti-microbial or
microbial-static agent, e.g. bacteriostatic agent,
preservative.
[0126] However, the production of preserved pharmaceutical
preparations containing protein or peptide actives has often proven
difficult, as when preservatives are used, these give rise to
stability problems. Often the proteins are inactivated and
aggregates are formed, which may sometimes lead to reported
injection site intolerance or immunogenicity to the active. This
can be further aggravated by additional excipients or formulation
components.
[0127] In one aspect each of the embodiments herein can optionally
contain an antimicrobial or microbial-static agent, which includes
bacteriostatic agents and preservative. Such agents include
benzalkonium chloride, m-cresol, benzyl alcohol or other phenolic
preservatives. Typical concentrations as known in the art can be
used.
[0128] However, surprisingly it has been found that the present
formulations with a peptide active agent do not require an
additional preservative, anti-microbial or microbial-static agent,
e.g. bacteriostatic or bacteriocide or additional amount of such
agent to provide a multi-use format. The formulations as described
herein provide a preservative effect with an acceptable peptide
stability and formulation stability. They can be used for
single-dose as well as for multiple-dose use. In this regard,
preferred formulations herein for multi-use format can contain
ethanol, propylene glycol, citric acid and/or EDTA as described,
preferably in sufficient concentrations to not only provide their
primary benefit as taught herein but also at sufficient
concentration, either alone or in any combination, to provide the
preservative effect while maintaining stability of the active and
the formulation.
[0129] Additional components above those mentioned as components a)
to f) will, where present at all, preferably be present in an
amount of 0 to 5% (e.g. 0.01% to 5%) by weight, preferably no more
than 2% by weight and more preferably no more than 1% by
weight.
[0130] In one embodiment, components a) and b) (allowing for any
impurity inherent in the nature of these components) make up at
least 95% of the lipid components of the composition. Preferably at
least 99% of the total lipid content of the pre-formulation
consists of components a) and b). Preferably the lipid component of
the pre-formulation consists essentially of components a) and
b).
Administration
[0131] The pre-formulations of the present invention are generally
formulated to be administered parenterally. This administration
will generally not be an intra-vascular method but will preferably
be subcutaneous (s.c.), intracavitary or intramuscular (i.m.).
Typically the administration will be by injection, which term is
used herein to indicate any method in which the formulation is
passed through the skin, such as by needle, catheter or needle-less
(needle-free) injector. It is, however, possible to take advantage
of the high loading and other beneficial characteristics of the
present formulation in non-parenteral applications, including
topical or systemic application to skin, mucous membranes, nasal,
buccal and/or oral cavities. Preferably, such non-parenteral
administration is for topical use.
[0132] Preferred parenteral administration is by i.m or s.c.
injection, most preferably by deep s.c. injection. An important
feature of the composition of the invention is that it can be
administered both by i.m. and s.c. and other routes without
toxicity or significant local effects. It is also suitable for
intracavital administration. The deep s.c. injection has the
advantage of being less deep and less painful to the subject than
the (deep) i.m. injection used for some current depots and is
technically most suitable in the present case as it combines ease
of injection with low risk of local side effects. It is a
surprising observation of the present inventors that the
formulations provide sustained release of active agent over a
predictable time period by both subcutaneous and intramuscular
injection. This therefore allows the site of injection to be varied
widely and allows the dose to be administered without detailed
consideration of the tissue depth at the site of injection.
[0133] The preferred lipid pre-formulations of the present
invention provide non-lamellar liquid crystalline depot
compositions upon exposure to aqueous fluids, especially in vivo.
As used herein, the term "non-lamellar" is used to indicate a
normal or reversed liquid crystalline phase (such as a cubic or
hexagonal phase) or the L3 phase or any combination thereof. The
term liquid crystalline indicates all hexagonal, all cubic liquid
crystalline phases and/or all mixtures thereof. Hexagonal as used
herein indicates "normal" or "reversed" hexagonal (preferably
reversed) and "cubic" indicates any cubic liquid crystalline phase
unless specified otherwise. The skilled reader will have no
difficulty in identifying those compositions having appropriate
phase behaviour by reference to the description and Examples
provided herein, and to WO2005/117830, but the most favoured
compositional area for phase behaviour is where ratio of components
a:b are in the region of 40:60 to 70:30, preferably 45:55 to 55:45
and more preferably 40:60 to 54:46. Ratios of around 50:50 (e.g.
.+-.2) are highly preferred, most preferably around 50:50.
[0134] It is important to appreciate that the pre-formulations of
the present invention are of low viscosity. As a result, these
pre-formulations must not be in any bulk liquid crystalline phase
since all liquid crystalline phases have a viscosity significantly
higher than could be administered by syringe or similar injecting
dispenser. The pre-formulations of the present invention will thus
be in a non-liquid crystalline state, such as a solution, L.sub.2
or L.sub.3 phase, particularly solution or L.sub.2. The L.sub.2
phase as used herein throughout is preferably a "swollen" L.sub.2
phase containing greater than 5 wt %, preferably greater than 7%,
and most preferably greater than 9% of organic mono-alcoholic
solvent (component c) having a viscosity reducing effect. The
preformulations of the invention which are in L.sub.2 phase form
one preferred set of preformulations and these will generally
contain at least 2% water as polar solvent.
[0135] As used herein, the term "low viscosity mixture" is used to
indicate a mixture which may be readily administered to a subject
and in particular readily administered by means of a standard
syringe and needle arrangement. This may be indicated, for example
by the ability to be dispensed from a 1 ml disposable syringe
through a small gauge needle. Preferably, the low viscosity
mixtures can be dispensed through a needle of 19 awg, preferably
smaller than 19 gauge, more preferably 23 awg (or most preferably
even 27 gauge) needle by manual pressure. In a particularly
preferred embodiment, the low viscosity mixture should be a mixture
capable of passing through a standard sterile filtration membrane
such as a 0.22 .mu.m syringe filter. A typical range of suitable
viscosities would be, for example, 0.1 to 5000 mPas, preferably 1
to 1000 mPas, more preferably 10 to 750 mPas and most preferably 25
to 500 mPas at 20.degree. C.
[0136] It has been observed that by the addition of small amounts
of low viscosity organic mono-alcoholic solvent, as indicated
herein, a very significant change in viscosity can be provided. For
example, the addition of only 5% solvent to a lipid mixture can
reduce viscosity 100-fold and addition of 10% may reduce the
viscosity up to 10,000 fold. In order to achieve this non-linear,
synergistic effect in lowering viscosity it is important that a
solvent of appropriately low viscosity and suitable polarity be
employed. Such solvents include those described herein infra.
Preferred low-viscosity mixtures include molecular solutions,
including dispersions of the peptide active agent in a molecular
solution of the other components.
[0137] Upon administration, the preferred lipid-based
pre-formulations of the present invention undergo a phase structure
transition from a low viscosity mixture to a high viscosity
(generally tissue adherent) depot composition. Generally this will
be a transition from a molecular mixture, swollen L.sub.2 and/or
L.sub.3 phase to one or more (high viscosity) liquid crystalline
phases such as normal or reversed hexagonal or cubic liquid
crystalline phases or mixtures thereof. Further phase transitions
may also take place following administration. Obviously, complete
phase transition is not necessary for the functioning of the
invention but at least a surface layer of the administered mixture
will form a liquid crystalline structure. Generally this transition
will be rapid for at least the surface region of the administered
formulation (that part in direct contact with air, body surfaces
and/or body fluids). This will most preferably be over a few
seconds or minutes (e.g. from 1 second up to 30 minutes, preferably
up to 10 minutes, more preferably 5 minutes of less). The remainder
of the composition may change phase to a liquid crystalline phase
more slowly by diffusion and/or as the surface region
disperses.
[0138] Without being bound by theory, it is believed that upon
exposure to excess aqueous fluid, the pre-formulations of the
invention lose some or all of the organic solvent included therein
(e.g. by diffusion) and take in aqueous fluid from the bodily
environment (e.g. the in vivo environment). For lipid
pre-formulations, at least a part of the formulation preferably
generates a non-lamellar, particularly liquid crystalline phase
structure. In most cases these non-lamellar structures are highly
viscous and are not easily dissolved or dispersed into the in vivo
environment. The result is a monolithic "depot" generated in vivo
with only a limited area of exposure to body fluids. Furthermore,
because the non-lamellar structure has large polar, apolar and
boundary regions, the lipid depot is highly effective in
solubilising and stabilising active agents such as peptides and
protecting these from degradation mechanisms. As the depot
composition formed from the pre-formulation gradually degrades over
a period of days, weeks or months, the active agent is gradually
released and/or diffuses out from the composition. Since the
environment within the depot composition is relatively protected,
the pre-formulations of the invention are highly suitable for
active agents with a relatively low biological half-life (see
above).
[0139] By incorporation of at least 10% of a polar solvent
(especially at least 5% water) into the pre-formulations, it is
believed that the rate of phase transition to a non-lamellar (e.g.
liquid crystalline) phase at the surface of the injected
pre-formulation can be enhanced in comparison with compositions
containing organic solvents in the substantial absence of water.
The performance of the resulting depot is thus improved and further
control over the release of active agent achieved.
[0140] The depot systems formed by the formulations of the present
invention are highly effective in protecting the active agent from
degradation and thus allow an extended release period. The
formulations of the invention thus may provide in vivo depots of
peptide active agents which require administration only once every
5 to 90 days preferably 5 to 60 days, more preferably 6 to 32.
Evidently, a longer stable release period is desirable for patient
comfort and compliance, as well as demanding less time from health
professionals if the composition is not to be self-administered.
Where the composition is to be self-administered, patient
compliance may be aided by a weekly (e.g. every 7 days, optionally
.+-.1 day) or monthly (e.g. every 28 or 30 days (optionally .+-.7
days) administration so that the need to administer is not
forgotten.
[0141] A considerable advantage of the depot precursors of the
present invention is that they are stable homogeneous phases. That
is to say, they may be stored for considerable periods (preferably
at least 6 months) at room or refrigerator temperature, without
phase separation. As well as providing advantageous storage and
facile administration, this allows for the dose of peptide active
agent (e.g. Somatostatin analogue, e.g. octreotide) to be selected
by reference to the species, age, sex, weight, and/or physical
condition of the individual subject, by means of injecting a
selected volume.
[0142] The present invention thus provides for methods comprising
the selection of a dosing amount specific to an individual,
particularly by subject weight. The means for this dose selection
is the choice of administration volume.
[0143] In one preferred aspect, the present invention provides a
pre-formulation comprising components a), b), c), d), f) and at
least one peptide active agent (e.g. somatostatin analogue, e.g.
octreotide) as indicated herein. The amounts of these components
will typically be in the range 30-70% a), 30-60% b), 5-20% c) and
0.1-20% d), with the peptide active agent (e.g. somatostatin
analogue, e.g. octreotide) present at 0.01% to 10%, (such as 36-44%
a), 36-44% b), 3-18% c) and 5-18% d) (preferably including at least
2% water), with the peptide active agent (e.g. somatostatin
analogue, e.g. octreotide) present at 1% to 3%), wherein the ratio
of a:b is in the range 40:60 to 54:46.
[0144] Typically, component f) is present at an antioxidant to
peptide active agent molar ratio of 1:50 to 1:1500, preferably
1:100 to 1:1300, and most preferably 1:150 to 1:1250. Since typical
antioxidants are of lower molecular weight than peptide active
agent (e.g. somatostatin analogue, e.g. octreotide), the proportion
by weight of antioxidant may be relatively small. For example, with
a small molecular weight pH adjuster (e.g. less than 500 amu),
0.001 to 5% of the composition may be antioxidant, preferably 0.002
to 2%, more preferably 0.002 to 0.15%, e.g. 0.002 to 0.015%.
[0145] The pre-formulations of the present invention are highly
advantageous in that they are stable to prolonged storage in their
final "administration ready" form. As a result, they may readily be
supplied for administration either by health professionals or by
patients or their carers, who need not be fully trained health
professionals and may not have the experience or skills to make up
complex preparations. This is particularly important in
long-duration, slow-effecting diseases such as diabetes.
Devices
[0146] In a yet further aspect, the present invention provides a
disposable administration device (which is also to include a device
component) pre-loaded with a measured dose of a pre-formulation of
the present invention. Such a device will typically contain a
single dose ready for administration, and will generally be
sterile-packed such that the composition is stored within the
device until administration. Suitable devices include cartridges,
ampoules and particularly syringes and syringe barrels, either with
integral needles or with standard (e.g. luer) fittings adapted to
take a suitable disposable needle.
[0147] The pre-filled devices of the invention may also suitably be
included in an administration kit, which kit also forms a further
aspect of the invention. In a still further aspect, the invention
thus provides a kit for the administration of at least one peptide
active agent, said kit containing a measured dose of a formulation
of the invention and optionally an administration device or
component thereof. Preferably the dose will be held within the
device or component, which will be suitable for i.m. or preferably
s.c. administration. The kits may include additional administration
components such as needles, swabs, etc. and will optionally and
preferably contain instructions for administration. Such
instructions will typically relate to administration by a route as
described herein and/or for the treatment of a disease indicated
herein above.
Kits
[0148] The invention provides for a pre-filled administration
device as indicated herein and a kit as indicated herein comprising
a pre-formulation as described herein.
[0149] In an alternative aspect of the present invention, the "kit"
may contain at least two vessels, a first containing a low
viscosity mixture of components a) to d), as described here, and a
second containing a measured dose of at least one peptide active
agent as described herein. The antioxidant component f) may be
formulated with the active agent, or more preferably as part of the
low viscosity mixture, which will then comprise components a) to d)
and f).
[0150] Such a "two component kit" may comprise the peptide active
agent as a powder formulation in one vial or pre-filled syringe and
components a) to d) (and optionally f)) in a second vial or
pre-filled syringe. In the case of two syringes, before injection,
the pre-filled syringes are connected and the powder comprising
active agent is mixed with the matrix formulation by moving the
syringe barrels back and forth, forming a solution or suspension
which is injected. Alternatively, the liquid lipid formulation is
drawn from one vial, or is pre-filled into a syringe, and is
injected into a vial containing peptide powder. This formulation
may subsequently be mixed by hand shaking or other suitable
reconstitution method (e.g. vortex mixing etc.). The solvent
component may be present in either or both vessels (e.g. vials or
syringes). Where the solvent is at least partially constituted with
the active agent, this will generally be in the form of a solution
or suspension.
[0151] In this aspect, the invention therefore provides a two
component kit comprising
i) a first vessel containing a low viscosity mixture of components
a) to d) as described herein; ii) a second vessel containing at
least one peptide active agent, iii) an antioxidant component f)
optionally in a third vessel, preferably in the second vessel, or
most preferably in the first vessel; iv) optionally and preferably
at least one of: [0152] 1) at least one syringe (which may be one
or both of said first and second vessels); [0153] 2) a needle for
administration, such as those described herein; [0154] 3)
instructions for generation of a composition of the invention from
the contents of the first and second vessels; [0155] 4)
instructions for administration, whereby to form a depot as
described herein.
Preferred Features and Combinations
[0156] In combination with the features and preferred features
indicated herein, the pre-formulations of the invention may have
one or more of the following preferred features independently or in
combination:
[0157] All proportions indicated herein may optionally be varied by
up to 10% of the amount specified, optionally and preferably by up
to 5%;
[0158] Component a) comprises, consists essentially of or
preferably consists of GDO;
[0159] Component b) comprises, consists essentially of or
preferably consists of soy PC;
[0160] Component c) comprises, consists essentially of or
preferably consists of a 1, 2, 3 or 4 carbon alcohol, preferably
isopropanol or more preferably ethanol;
[0161] Component d) comprises, consists essentially of or
preferably consists of a polar solvent such as water, propylene
glycol, or mixtures thereof;
[0162] Component f) comprises, consists essentially of or
preferably consists of ascorbic acid, ethylenediaminetetraacetic
acid (EDTA), and/or citric acid;
[0163] The pre-formulation contains at least one peptide active
agent, preferably a Somatostatin analogue such as Octreotide;
[0164] The pre-formulation contains at least one somatostatin
analogue (as described herein) such as at least one peptide which
has agonistic and/or antagonistic effect at at least one of the
SST(1)-SST(5) receptors (e.g. in humans).
[0165] The pre-formulation does not contain any somatostatin
analogue (as described herein); The pre-formulation has a low
viscosity as indicated herein.
[0166] The pre-formulation comprises forms a liquid crystalline
phase as indicated herein upon in vivo administration.
[0167] The pre-formulation generates a depot following in vivo
administration, which depot releases at least one active agent at a
therapeutic level over a period of at least 7 days, preferably at
least 21 days, more preferably at least 28 days.
[0168] The pre-formulation has a higher loading of peptide active
agent (e.g. Somatostatin analogue, e.g. octreotide) than is stable
in the same formulation in the absence of component e).
[0169] The pre-formulation has a higher loading of peptide active
agent (e.g. Somatostatin analogue, e.g. octreotide) than is
obtainable by equilibration at 25.degree. C. of the same
formulation in the absence of component f).
[0170] In combination with the features and preferred features
indicated herein, the method(s) of treatment of the present
invention may have one or more of the following preferred features
independently or in combination:
[0171] The method comprises the administration of at least one
formulation with one or more preferred features as indicated
above;
[0172] The method comprises the administration of at least one
formulation as indicated herein by i.m., s.c. or preferably deep
s.c. injection;
[0173] The method comprises administration by means of a pre-filled
administration device as indicated herein;
[0174] The method comprises administration through a needle no
larger than 20 gauge, preferably smaller than 20 gauge, and most
preferably 23 gauge or smaller;
[0175] The method comprises a single administration every 5 to 90
days, preferably 6 to 32 days (for example 7 days or 28-31
days).
[0176] In combination with the features and preferred features
indicated herein, the use(s) of the pre-formulations indicated
herein in the manufacture of medicaments may have one or more of
the following preferred features independently or in
combination:
[0177] The use comprises the use of at least one formulation with
one or more preferred features as indicated above;
[0178] The use comprises the manufacture of a medicament for
administration of at least one formulation as indicated herein by
i.m., s.c. or preferably deep s.c. injection;
[0179] The use comprises the manufacture of a medicament for
administration by means of a pre-filled administration device as
indicated herein;
[0180] The use comprises the manufacture of a medicament for
administration through a needle no larger than 20 gauge, preferably
smaller than 20 gauge, and most preferably 23 gauge or smaller;
[0181] The use comprises the manufacture of a medicament for
administration once every 5 to 90 days, preferably 5 to 60 days,
more preferably 6 to 32 days.
[0182] In combination with the features and preferred features
indicated herein, the pre-filled devices of the invention may have
one or more of the following preferred features independently or in
combination:
[0183] They contain a preferred formulation as indicated
herein;
[0184] They comprise a needle smaller than 20 gauge, preferably no
larger than 23 gauge;
[0185] They contain a single dose of 1 to 2000 mg of peptide active
agent (e.g. Somatostatin analogue, e.g. octreotide), preferably 0.1
to 100 mg and more preferably 1-50 mg, most preferably 5-35 mg
[0186] They contain peptide active agent Somatostatin analogue
(e.g. octreotide or exenatide) at around 1 to 100 mg.
[0187] They contain a homogeneous mixture of a composition of the
invention in ready-to-inject form.
[0188] They contain a formulation of components a) to c) for
combination with a peptide active agent whereby to form a
preformulation of the invention.
[0189] They contain a peptide active agent for combination with a
formulation of components a) to c) and optionally e), whereby to
form a preformulation of the invention.
[0190] They contain a total volume for administration of no more
than 5 ml, preferably no more than 3 ml more preferably no more
than 1.5 ml.
[0191] In combination with the features and preferred features
indicated herein, the kits of the invention may have one or more of
the following preferred features independently or in
combination:
[0192] They contain a preferred formulation as indicated
herein;
[0193] They contain a pre-filled device as indicated herein;
[0194] They contain a needle smaller than 20 gauge, preferably no
larger than 23 gauge;
[0195] They contain a single dose of 1 to 200 mg of peptide active
agent (e.g. Somatostatin analogue, e.g. octreotide), preferably 1
to 100 mg and more preferably 1-50 mg;
[0196] They contain peptide active agent. Somatostatin analogue,
e.g. octreotide, at around 1 to 100 mg;
[0197] They contain a "two compartment kit" comprising at least two
vessels containing a lipid formulation of the invention and a
peptide active agent (e.g. Somatostatin analogue, e.g. octreotide)
powder, respectively.
[0198] They contain a total volume for administration of no more
than 5 ml, preferably no more than 3 ml more preferably no more
than 1.5 ml.
[0199] They contain instructions for administration by a route
and/or at a frequency as indicated herein;
[0200] They contain instructions for administration for use in a
method of treatment as described herein.
[0201] The Invention will now be further illustrated by reference
to the following non-limiting Examples and the attached
Figures.
EXAMPLES
TABLE-US-00009 [0202] Abbreviations OCT(Cl) Octreotide
hydrochloride (PolyPeptide Labs., USA) SOM(Ac) Somatostatin 1-14
acetate (PolyPeptide Labs., USA) LEU(Ac) Leuprolide acetate
(PolyPeptide Labs., USA) TTA Triptorelin acetate (Bachem,
Switzerland) TTP Triptorelin pamoate (Bachem, Switzerland) SPC Soy
phosphatidylcholine (Lipoid, Germany) GDO Glycerol dioleate
(Danisco, Denmark) DOPC Dioleoyl phosphatidylcholine (NOF, Japan)
EtOH Ethanol (99.5 vol %, Ph. Eur., USP) PG Propylene glycol (Ph.
Eur., USP)
Example 1: Manufacturing of OCT-Containing Products
TABLE-US-00010 [0203] TABLE 1 Composition of OCT-containing
products. Formu- EDTA- lation Ingredient OCT(Cl) SPC GDO EtOH PG
water .sup.1) A (wt %) 2.44 43.78 43.78 5.00 5.00 -- B (wt %) 2.44
42.28 42.28 6.50 6.50 -- C (wt %) 2.44 45.53 45.53 6.50 -- -- D (wt
%) 2.44 38.78 38.78 10.00 -- 10.00 E (wt %) 2.44 33.78 33.78 15.00
-- 15.00 .sup.1) The concentration in the solution is 0.10 mg
EDTA/mL; this solution was prepared by mixing 10 mg EDTANa.sub.2 in
water.
[0204] Depot precursors with the compositions presented in Table 1
were manufactured by first preparing a peptide stock, by weighing
the ingredients as described in Table 2 and mixing on a shaking
table (250-300 rpm) to homogeneous solutions.
TABLE-US-00011 TABLE 2 Preparation of peptide stock solutions.
Formulation Ingredient OCT(Cl) EtOH PG EDTA-water A (g) 2.44 5.00
5.00 -- B (g) 2.44 6.50 6.50 -- C (g) 2.44 6.50 -- -- D (g) 2.56 --
-- 10.50 E (g) 0.28 -- -- 1.73
[0205] For manufacturing of A, B and C, the following amounts
(Table 3) of SPC and GDO were weighted directly into the recipient
containing the OCT(Cl) stock solution.
TABLE-US-00012 TABLE 3 Amounts of SPC and GDO added for preparation
of CAM2029-BP, -BR, and -BU. Formulation Ingredient SPC GDO A (g)
43.78 43.78 B (g) 42.28 42.28 C (g) 45.53 45.53
[0206] The mixtures were then placed on a shaking table (250-300
rpm) until homogeneous solutions were obtained.
[0207] For manufacturing of formulation D, a lipid stock was
prepared by mixing (shaking table (250-300 rpm)) 88.58 g SPC, 22.84
g EtOH and 88.58 g GDO to a homogeneous solution. The final
formulation was then obtained by combining 87.56 g lipid stock and
12.44 g peptide stock solution and mixing (shaking table (250-300
rpm)) to homogeneous.
[0208] For manufacturing of formulation E, a lipid stock was
prepared by mixing (shaking table (250-300 rpm)) 4.09 g SPC, 1.82 g
EtOH and 4.09 g GDO to a homogeneous solution. The final
formulation was then obtained by combining 8.26 g lipid stock and
1.74 g peptide stock solution and mixing (shaking table (250-300
rpm)) to homogeneous.
Example 2: In-Vitro Release from OCT-Containing Products
[0209] Formulations with the composition presented in Table 4 were
manufactured by first preparing the corresponding OCT(Cl) stock
solutions in EtOH, EtOH:PG mixture or respectively water (as
described in Example 1 above), followed by mixing with the other
components until homogeneous solutions were obtained.
TABLE-US-00013 TABLE 4 Composition of OCT-containing products
evaluated in the accelerated in vitro release (IVR) experiment.
Formu- lation Ingredient OCT(Cl) SPC GDO EtOH PG Water 911 (wt %)
2.43 43.72 43.79 5.02 5.04 912 (wt %) 2.43 42.08 42.13 6.56 6.81
913 (wt %) 2.43 41.26 41.23 7.51 7.58 914 (wt %) 2.43 45.41 45.60
6.56 916 (wt %) 2.43 33.70 33.73 15.09 15.06 917 (wt %) 2.44 38.70
38.68 10.04 10.14 918 (wt %) 3.64 33.08 33.07 15.08 15.13 1006 (wt
%) 2.00 41.48 41.48 7.55 7.49 1007 (wt %) 2.00 36.49 36.49 12.50
12.53 1010 (wt %) 2.00 38.96 38.95 10.12 9.97
[0210] Evaluation of accelerated IVR of OCT from each of the
formulations presented above was carried out by injecting
approximately 100 mg (.+-.20%) into a glass vial containing 5 mL of
phosphate buffered saline:acetonitrile 85:15 (v/v) mixture. The
vials were sealed, and incubated at room temperature for up to 48
h. Sampling was carried out at different time points from the
initiation of the experiment, by slowly pulling out 0.2 mL of the
aqueous phase, which was collected directly into a 0.3 mL HPLC
polypropylene vial. The analysis was performed by HPLC-UV using an
analytical column (ACE-5 C18, 50.times.3.0 mm) with gradient
elution (mobile phase A: 0.1 vol. % trifluoroacetic acid (TFA) in
water; mobile phase B: 0.1 vol. % in 95 vol. % methanol, 5 vol. %
water) and UV detection at 282 nm.
[0211] The results obtained are presented in FIGS. 1 (a and b)
Example 3: Stability of OCT-Containing Products with Vs. Without
EDTA
[0212] A formulation (batch size 110 g) with the composition
OCT(Cl)/SPC/GDO/EtOH 3.74/43.13/43.13/10.00 (all in wt %) was
manufactured by first dissolving 4.114 g OCT(Cl) in 11.000 g EtOH,
followed by sequential addition of 47.433 g SPC and 47.433 g GDO,
and mixing to a homogeneous solution (91).
[0213] One sample (G) containing 3.37 wt % OCT(Cl) (approximately
2.98 wt % OCT base) by mixing 0.9 g formulation 91 with 0.1 mg of a
solution containing 0.1 wt % of EDTA in HPLC-grade water.
[0214] One sample (H) containing 3.37 wt % OCT(Cl) was prepared by
mixing 0.9 g formulation 91 with 0.1 mg HPLC-grade water.
[0215] The samples were divided each into two aliquots of about 0.4
g/vial; one aliquot/sample was incubated at 70.degree. C., whereas
the other was placed at <-15.degree. C. (reference). All samples
were analysed after 7 days of incubation in the above-mentioned
conditions by using a normal-phase HPLC (analytical column
LiChrospher Diol 5 .mu.m, 250.times.3.2 mm) UV/DAD--based
analytical method for quantification of OCT and relative
determination of OCT-related substances. The presence of EDTA in
the water phase considerably improved the stability of OCT in the
lipid matrix, as shown in the results presented in FIG. 2.
Example 4: Injectability of OCT-Containing Products
[0216] The injectability is here defined as the flow rate of the
evaluated fluid from a syringe (specified by its volume and design)
through a needle (specified by its needle gauge and length)
subjected to a constant force against atmospheric pressure.
[0217] For filling purposes, a thick needle was preferred, e.g. an
18 G needle. When the syringe was filled with the necessary amount
of formulation, the thick needle was exchanged for the needle to be
examined. By pressing on the plunger with the new needle in place,
the entrapped air was removed. The excess formulation was wiped off
with a paper tissue and the starting weight (grams) of the filled
syringe was measured. The syringe was then mounted in a vertical
position using a metallic stand with holder and with the needle
facing down. The ejected fluid was collected directly in a glass
vial.
[0218] A 20N weight was placed centered on the plunger and the
timer was started when the weight and the plunger come into
contact. The time to empty the syringe (seconds) was then
monitored. After the syringe has been emptied, its final weight
(grams) was measured. At least two repeat measurements for each
sample and type of needle were performed.
[0219] The injectability was calculated by use of the following
equation:
Injectability = ( Starting .times. .times. weight - Final .times.
.times. weight ) Injection .times. .times. time .times. ( mg s )
##EQU00001##
[0220] The injectability of several OCT-containing products is
presented in Table 5.
TABLE-US-00014 TABLE 5 Injectability (mg formulation/s) of
OCT-containing products through 23 G thin-wall (Terumo Neolus
NN-2316R), respective 25 G thin-wall (Terumo Neolus NN-2516R) 16
mm-long needles. The syringes used were BD 1 mL plast Luer-Lock
(#309628). Injectability (mg/s) Injectability (mg/s) Formulation
through 23 G needle through 25 G needle 91 71 25 911 49 17 912 86
38 913 126 49 914 32 12 916 252 109 917 126 49 918 308 115
Example 5: In Vivo PK Studies in Rats
Animals and Source
[0221] Male SPF Sprague-Dawley rats (NTAC:SD) from M&B Taconic
Europe A/S (Ejby, Denmark) were used in the studies. At arrival the
rats were 8 to 9 weeks old, with a bodyweight in the range from 275
to 300 g. An acclimatization period of at least 5 days was allowed
before dosing.
Housing
[0222] The rats will be kept in pairs in transparent polycarbonate
cages (Macrolon.RTM. type III; Scanbur BK A/S, Karlslunde, Denmark)
with a floor area of 810 cm.sup.2. Aspen wooden chopping (Tapvei
Aspen Bedding, Tapvei Oy, Kortteinen, Finland) were used for
bedding material. Wood wool for nest building (PM 90 L
"Bobyggnadsmaterial", Tapvei) and a piece of wood ("Gnagpinne
medium", Tapvei) were used as environmental enrichment. Complete
pelleted rodent diet (Labfor R70, Kimstad, Sweden) and water were
available ad libitum.
Dosing
[0223] The animals were dosed according to Camurus internal
standard operating procedure (SOP PK12-3). In brief, dosing was
performed by subcutaneous injections between the scapulae under
light isofluran anesthesia, using a 1-mL Luer-lock syringe and a
25-mm 23 G needle.
Blood Sampling
[0224] Blood samples were collected from awake animals by
sub-lingual bleeding. Sampling time points was pre-dose, and 1
hour, 6 hours, 1 day, 2 days, 5 days, 8 days, 14 days, 21 days, 28
days and 35 days after dosing. Blood was collected into
EDTA-treated test tubes (Capiject 3T-MQK, Terumo Medical
Corporation), placed on ice immediately after collection. After
centrifugation (approximately 1500.times.g, at 5.degree. C. for 10
min) the plasma was transferred new test tubes and stored below
-70.degree. C. until analysis.
Bioanalysis
[0225] Analysis of OCT--The plasma samples was analysed with the
ELISA kit S-1275 (Bachem/Peninsula Laboratories) "Octreotide--EIA
Kit, Host: Rabbit, High Sensitivity", adapted for analysis of OCT
in rat EDTA plasma.
Results
[0226] Pharmacokinetic (PK) profiles for formulations 91, 911 and
912 are shown in FIG. 3.
Example 6: Manufacturing of Formulations Containing Leuprolide
Acetate (Leuprorelin-LEU(Ac))
[0227] Leuprolide compositions according to the invention were
prepared with the compositions as indicated in Table 6. The
formulations were prepared by first dissolving the LEU(Ac) in the
EtOH, WFI and/or PG components, whereafter the lipid components
were added sequentially, starting with SPC and followed by GDO. The
final formulations were mixed on a shaking table at 250-300 rpm
until clear and homogenous liquid solutions were obtained. The
formulations were finally subjected to sterile filtration (0.2
.mu.m sterile PVDF filter from Millipore) under 2 bar nitrogen
pressure.
TABLE-US-00015 TABLE 6 Composition (wt %) of leuprolide acetate
(LEU(Ac)) formulations. Formulation# LEU(Ac) SPC DOPC GDO EtOH WFI
PG 49 2.70.sup.1) 43.65 -- 43.65 10.00 -- -- 50 2.70.sup.1) 37.65
-- 37.65 12.00 10.00 -- 51 1.62.sup.2) 38.19 -- 38.19 12.00 10.00
-- 52 2.70.sup.1) 33.65 -- 33.65 15.00 15.00 -- 53 2.70.sup.1)
42.15 -- 42.15 6.50 -- 6.50 54 2.70.sup.1) 41.15 -- 41.15 7.50 --
7.50 55 2.70.sup.1) 38.65 -- 38.65 10.00 -- 10.00 56 1.62.sup.2)
41.69 -- 41.69 7.50 -- 7.50 57 2.70.sup.1) -- 41.15 41.15 7.50 --
7.50 .sup.1)Corresponding to 25 mg leuprolide acetate per mL when
corrected for peptide purity and content and formulation density.
.sup.2)Corresponding to 15 mg leuprolide acetate per mL when
corrected for peptide purity and content and formulation
density.
Example 7: Manufacturing of Formulations Containing Triptorelin
Acetate (TTA) and Triptorelin Pamoate (TTP)
[0228] Triptorelin acetate and pamoate compositions according to
the invention were prepared with the compositions as indicated in
Table 7. The formulations were prepared by first mixing the TTA or
TTP in the EtOH and PG components, whereafter the lipid components
were added sequentially, starting with SPC and followed by GDO. The
final formulations were mixed on a shaking table at 250-300 rpm
until clear and homogenous liquid solutions were obtained. The
formulations were finally subjected to sterile filtration (0.2
.mu.m sterile PVDF filter from Millipore) under 2 bar nitrogen
pressure.
TABLE-US-00016 TABLE 7 Composition (wt %) of triptorelin acetate
(TTA) and triptorelin pamoate (TTP) formulations. Formulation# TTA
TTP SPC GDO EtOH PG 58 3.00.sup.1) -- 41.00 41.00 7.50 7.50 59
3.00.sup.1) -- 38.50 38.50 10.00 10.00 60 -- 3.50.sup.1) 40.75
40.75 7.50 7.50 61 -- 3.50.sup.1) 38.25 38.25 10.00 10.00
.sup.1)Corresponding to 25 mg triptorelin free base per mL when
corrected for peptide purity and content and formulation
density.
Example 8: Manufacturing of Further Formulations Containing
Octreotide Chloride (OCT(Cl))
[0229] Octreotide compositions according to the invention were
prepared with the compositions as indicated in Table 8. The
formulations were prepared by first dissolving the OCT(Cl) in the
EtOH, WFI and/or PG components, whereafter the lipid components
were added sequentially, starting with SPC and followed by GDO. The
final formulations were mixed on a shaking table at 250-300 rpm
until clear and homogenous liquid solutions were obtained. The
formulations were finally subjected to sterile filtration (0.2
.mu.m sterile PVDF filter from Millipore) under 2 bar nitrogen
pressure.
TABLE-US-00017 TABLE 8 Composition (wt %) of octreotide chloride
(OCT(Cl)) formulations. Formulation# OCT(Cl) SPC GDO EtOH WFI 81
(91) 3.65.sup.1) 43.18 43.18 10 -- 82 2.44.sup.2) 43.78 43.78 10 --
83 (917) 2.44.sup.2) 38.78 38.78 10 10 84 1.46.sup.3) 39.27 39.27
10 10 Corresponding to .sup.1)30 mg, .sup.2)20 mg, and .sup.3)12 mg
octreotide free base per mL when corrected for peptide purity and
content and formulation density.
Example 9: Manufacturing of Formulations Containing Somatostatin
1-14 Acetate (SOM(Ac)) and Somatostatin 1-14 Hydrochloride
(SOM(Cl))
[0230] Somatostatin (1-14) acetate (SOM(Ac)) and hydrochloride
(SOM(Cl)) compositions according to the invention were prepared
with the compositions as indicated in Table 9. The hydrochloride
salt, SOM(Cl), was prepared from the acetate salt via an
ion-exchange chromatography process followed by lyophilisation of
the peptide solution by freeze-drying. Complete counter-ion
exchange was confirmed by HPLC. The formulations were prepared by
first mixing the lipid components, SPC and GDO, with the EtOH and
PG components, followed by mixing on a shaking table at 250-300 rpm
to form homogenous lipid solutions. To the lipid solution, the
respective SOM(Ac) and SOM(Cl) drug powders were added in the
required amount. The final formulations were mixed by end-over-end
rotation at ambient room temperature until clear and homogenous
liquid solutions were obtained. The formulations were finally
subjected to sterile filtration (0.2 .mu.m sterile PVDF filter from
Millipore) under 2 bar nitrogen pressure.
TABLE-US-00018 TABLE 9 Composition (wt %) of somatostatin 1-14
acetate (SOM(Ac)) and somatostatin 1-14 hydrochloride (SOM(Cl))
formulations. Formulation# SOM(Ac) SOM(Cl) SPC GDO EtOH PG 9 --
3.00 43.50 43.50 5 5 11 -- 4.00 38.00 38.00 10 10 14 3.00 -- 38.50
38.50 10 10
Example 10: In Vivo Studies of Leuprolide Formulations in Rats
[0231] For general aspects, see Example 5. Dosing of the rats was
performed by subcutaneous injection of formulation 49 and 50,
respectively (see Table 6).
Blood Samples for Pharmacokinetics
[0232] Blood for pharmacokinetics were collected pre-dose, and 1
hour, 4 hours, 10 hours, 1 day, 2 days, 3 days, 5 days, 7 days, 14
days and 21 days after dosing. The factual time points for sampling
were calculated as the difference between time for sampling and
time of dosing. A deviation of .+-.10% from the nominal time was
accepted.
Bioanalysis
[0233] Analysis of Leuprolide was performed using the (Des-Gly10,
D-LEU6, Pro-NHEt9)-LHRH (Leuprolide) high sensitivity EIA kit
(S-1174, Bachem/Peninsula Laboratories) adapted for analysis of LEU
in rat EDTA plasma.
Results
[0234] Pharmacokinetic (PK) profiles for formulations 49 and 50 are
shown in FIG. 4.
Sequence CWU 1
1
2114PRTHomo sapiens 1Ala Gly Cys Lys Asn Phe Phe Trp Lys Thr Phe
Thr Ser Cys1 5 1024PRTHomo sapiens 2Phe Trp Lys Thr1
* * * * *