U.S. patent application number 15/794537 was filed with the patent office on 2018-07-26 for acid containing lipid 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 | 20180207089 15/794537 |
Document ID | / |
Family ID | 38599087 |
Filed Date | 2018-07-26 |
United States Patent
Application |
20180207089 |
Kind Code |
A1 |
NISTOR; Catalin ; et
al. |
July 26, 2018 |
ACID CONTAINING LIPID FORMULATIONS
Abstract
The present invention relates to compositions forming a low
viscosity mixture of: i) a non-polymeric slow-release matrix ii) at
least one biocompatible, (preferably oxygen containing) organic
solvent; iii) at least one peptide active agent; and iv) at least
one lipid soluble acid. The invention further relates to methods of
treatment comprising administration of such compositions,
especially in treating diabetes, and to pre-filled administration
devices and kits containing the formulations.
Inventors: |
NISTOR; Catalin; (Lund,
SE) ; JOHNSSON; Markus; (Lund, SE) ; TIBERG;
Fredrik; (Lund, SE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
CAMURUS AB |
Lund |
|
SE |
|
|
Assignee: |
CAMURUS AB
Lund
SE
|
Family ID: |
38599087 |
Appl. No.: |
15/794537 |
Filed: |
October 26, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12674226 |
Jun 2, 2011 |
9820934 |
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PCT/GB2008/002857 |
Aug 22, 2008 |
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15794537 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 9/1274 20130101;
A61K 9/0019 20130101; A61P 3/10 20180101; A61K 47/14 20130101; A61K
9/127 20130101; A61K 9/06 20130101; A61K 47/24 20130101; A61K 38/26
20130101; A61K 9/0024 20130101; A61K 47/10 20130101 |
International
Class: |
A61K 9/00 20060101
A61K009/00; A61K 9/127 20060101 A61K009/127; A61K 9/06 20060101
A61K009/06; A61K 47/14 20060101 A61K047/14; A61K 47/10 20060101
A61K047/10; A61K 38/26 20060101 A61K038/26; A61K 47/24 20060101
A61K047/24 |
Foreign Application Data
Date |
Code |
Application Number |
Aug 22, 2007 |
GB |
0716385.0 |
Claims
1-21. (canceled)
22. A non-aqueous pre-formulation comprising a low viscosity
mixture of: i) a lipid-based slow-release matrix; ii) at least one
biocompatible, oxygen-containing organic solvent selected from the
group consisting of: an alcohol, a ketone, an ester, an ether, an
amide or a sulfoxide; iii) at least one peptide active agent; and
iv) at least one lipid soluble acid.
23. A non-aqueous pre-formulation as claimed in claim 22 comprising
a low viscosity mixture of: a) at least one neutral diacyl lipid
and/or a tocopherol; b) at least one phospholipid; c) at least one
biocompatible, oxygen-containing organic solvent selected from the
group consisting of: an alcohol, a ketone, an ester, an ether, an
amide or a sulfoxide; d) at least one peptide active agent; and e)
at least one lipid soluble acid; wherein the pre-formulation forms,
or is capable of forming, at least one liquid crystalline phase
structure upon contact with an aqueous fluid.
24. A non-aqueous pre-formulation as claimed in claim 22 comprising
a low viscosity mixture of: a) at least one diacyl glycerol; b) at
least one phosphatidyl choline; c) at least one oxygen-containing
organic solvent selected from the group consisting of: an alcohol,
a ketone, an ester, an ether, an amide or a sulfoxide; d) at least
one peptide active agent; and e) at least one lipid soluble acid;
wherein the pre-formulation forms, or is capable of forming, at
least one liquid crystalline phase structure upon contact with an
aqueous fluid.
25. A non-aqueous pre-formulation as claimed in claim 23 wherein
component a) is present at a level of 30-70% by weight.
26. A non-aqueous pre-formulation as claimed in claim 23 wherein
component b) is present at a level of 30-60% by weight.
27. A non-aqueous pre-formulation as claimed in claim 22 wherein
the oxygen-containing organic solvent is present at a level of 0.1
to 20% by weight.
28. A non-aqueous pre-formulation as claimed in claim 22 wherein
said lipid soluble acid is selected from the group consisting of:
benzoic acid, citric acid, a sulfonic acid or a hydrohalic
acid.
29. A non-aqueous pre-formulation as claimed in claim 22 wherein
said lipid soluble acid is selected from the group consisting of:
benzoic acid, citric acid, methane sulfonic acid, benzene sulfonic
acid, toluene sulfonic acid and HCl.
30. A non-aqueous pre-formulation as claimed in claim 22 wherein
said biocompatible, oxygen-containing organic solvent includes at
least one solvent selected from the group consisting of: a ketone,
an ester, an ether, an amide and a sulfoxide.
31. A non-aqueous pre-formulation as claimed in claim 22 wherein
said biocompatible, oxygen-containing organic solvent includes at
least one amide selected from the group consisting of: N-methyl
pyrrolidone (NMP), 2-pyrrolidone and dimethylacetamide (DMA).
32. A non-aqueous pre-formulation as claimed in claim 22 wherein
said biocompatible, oxygen-containing organic solvent includes
dimethylsulfoxide (DMSO).
33. A non-aqueous pre-formulation as claimed in claim 22 which is
capable of being dispensed through a needle of 19 awg by manual
pressure.
34. A method of delivery of a peptide active agent to a human or
non-human animal body, said method comprising parenterally
administering a non-aqueous pre-formulation comprising a low
viscosity mixture of: i) a non-polymeric slow-release matrix ii) at
least one biocompatible, oxygen-containing organic solvent selected
from the group consisting of: an alcohol, a ketone, an ester, an
ether, an amide or a sulfoxide; iii) at least one peptide active
agent; and iv) at least one lipid soluble acid.
35. A pre-filled administration device containing a pre-formulation
as claimed in claim 22.
36. A kit comprising an administration device as claimed in claim
35.
Description
[0001] The present invention relates to formulation precursors
(pre-formulations) for the in situ generation compositions for the
controlled release of peptide active agents such as
Glucagon-like-peptide-1 (GLP-1) and/or analogues thereof, and
methods of treatment with such formulations. In particular, the
invention relates to high-loading pre-formulations of amphiphilic
components and at least one GLP-1 or analogues active agent for
parenteral application, which undergo phase transition upon
exposure to aqueous fluids, such as body fluids, thereby forming a
controlled release matrix.
[0002] 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.
[0003] 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.
[0004] 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 wellbeing 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.
[0005] 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. the
carboxypeptidases A, B & C) 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 decrease their
bioavailability. Similarly, free peptides and proteins in the
mammalian blood stream are also subject to enzymic degradation
(e.g. by plasma carboxy peptidases etc.).
[0006] There are many peptide based active agents, some of which
are discussed herein below. Among these, one of particular interest
is GLP-1.
[0007] Glucagon-like peptide (GLP)-1 is a potent glucoregulatory
hormone that is released from intestinal L cells into the
circulation in response to nutrient ingestion and neural and
endocrine stimuli. Structurally, GLP-1 precursor (precursor to the
active forms) is a 37-amino acid peptide with a MW of 4.2 KDa,
having a sequence highly conserved between different species. After
post-translational cleavage of the first six amino acids of the
precursor, two equipotent active forms of GLP-1 ((7-37) and
(7-36)amide) are generated. GLP-1 is involved in modification of
glucose homeostasis through actions that include potentiation of
glucose-stimulated insulin secretion and biosynthesis and
suppression of glucagon secretion, gastric emptying, and food
intake. The abilities of GLP-1 to stimulate insulin secretion and
inhibit glucagon release are glucose-dependent; thus, the risk of
hypoglycemia with GLP-1 administration is low. GLP-1 also increases
beta-cell mass in preclinical models of diabetes through mechanisms
that include stimulation of beta-cell proliferation and neogenesis
and inhibition of beta-cell apoptosis. Studies in both animals and
humans indicate that GLP-1 may also play a protective role in the
cardiovascular system.
[0008] The combined actions of GLP-1 have generated substantial
interest in using this peptide as a therapeutic agent for the
treatment metabolic diseases, including type II diabetes and
obesity. However, the therapeutic potential of native GLP-1 is
limited by its very short plasma half-life (below 2 minutes). This
is due to both rapid inactivation by the proteolytic enzyme
dipeptidyl peptidase (DPP)-IV and renal clearance. Consequently,
long-acting, DPP-IV-resistant GLP-1 analogues have been developed
for clinical use, including exenatide (Byetta, Amylin-Lilly),
liraglutide (Novo Nordisk), CJC-1131 (ConjuChem), AVE010 (Zealand
Pharma--Sanofi-Aventis), LY548806 (Lilly), TH-0318
(TheraTechnologies), BIM 51077 (Ipsen-Roche). All these are once-
or twice-daily administration products; a controlled-release (one
week) exenatide product (Exenatide LAR Alkermes-Amylin-Lilly) is
currently under clinical investigation. These GLP-1 mimetics bind
to GLP-1 receptors with similar or higher affinity and produce
biological actions identical to those of native GLP-1 but are
resistant to DPP-IV-mediated inactivation and renal clearance.
These compounds are able to exert more sustained GLP-1-like
activity for longer periods of time in vivo. An alternative
therapeutic approach for prolonging the action of native GLP-1 is
to inhibit DPP-IV activity, thereby preventing GLP-1 degradation.
Several orally active agents that inhibit DPP-IV activity are also
being evaluated for the treatment of type II diabetes.
[0009] The structures and sequences of GLP-1 and some known
analogues are shown below starting with two equipotent naturally
occurring forms. A straightforward system is used to describe
fragments and analogues of GLP-1. For example,
Arg.sup.34-GLP-1(7-37) designates an analogue of GLP-1 formally
derived from GLP-1 precursor by deleting the amino acid residues
Nos. 1 to 6 and substituting the naturally occurring amino acid
residue in position 34 (Lys) by Arg.
[0010] Native (human) GLP-1(7-37):
[0011]
His.sup.7-Ala-Glu-Gly.sup.10-Thr-Phe-Thr-Ser-Asp.sup.15-Val-Ser-Ser-
-Tyr-Leu.sup.20-Glu-Gly-Gln-Ala-Ala.sup.25-Lys-Glu-Phe-Ile-Ala.sup.30-Trp--
Leu-Val-Lys-Gly-Arg-Gly.sup.37
[0012] Native (human):
[0013] GLP-1(7-36)amide
[0014] NovoNordisk (Liraglutide)
[0015]
Arg.sup.34Lys.sup.26-(N-.epsilon.-(.gamma.-Glu(N-.alpha.-hexadecano-
yl)))-GLP-1(7-37)
[0016] Conjuchem (CJC-1131)
[0017]
D-Ala.sup.8Lys.sup.37-(2-(2-(2-maleimidopropionamido(ethoxy)ethoxy)-
acetamide))-GLP-1(7-37)
[0018] Sanofi-Aventis/Zealand (AVE-010 (ZP10))
[0019]
His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln-Met-Glu-Glu-Glu-
-Ala-Val-Arg-Leu-Phe-Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-Pro-Ser-Ser-Gly-Ala-P-
ro-Pro-Ser-Lys-Lys-Lys-Lys-Lys-Lys
[0020] Eli Lilly (Exenatide)
[0021]
His.sup.7-Gly-Glu-Gly.sup.10-Thr-Phe-Thr-Ser-Asp.sup.15-Leu-Ser-Lys-
-Gln-Met.sup.20-Glu-Glu-Glu-Ala-Val.sup.25-Arg-Leu-Phe-Ile-Glu.sup.30-Trp--
Leu-Lys-Asn-Gly-Gly-Pro.sup.37-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-amide
[0022] As used herein, "native GLP-1" indicates human GLP-1(7-37)
and/or human GLP-1 (7-36)amide and the terms "Liraglutide",
"CJC-1131", "AVE-010", "exenatide" are used to indicate the
respective actives above, including their physiologically
acceptable salts, esters and derivatives where context allows. All
of these, including the native GLP-1 sequences are included in the
term "GLP-1 analogues" as used herein. Other suitable GLP-1
analogues are described in e.g. Knudsen et al. J. Med. Chem. 2000,
43, 1664-1669; Knudsen J. Med. Chem. 2004, 47, 4128-4134; Hui et
al. Diabetes Metab. Res. Rev. 2005, 21, 313-331 and Holz and
Chepumy Curr. Med. Chem. 2003, 10, 2471-2483. These citations are
incorporated herein by reference in their entireties, and although
specific passages are referred to herein, all GLP-1 analogue
sequences and all GLP-1 receptor agonists referred to in any of
these documents are suitable for use in the present invention.
GLP-1 receptor agonists as referred to herein includes all GLP-1
analogues as described above and in the references cited above.
[0023] With regard to administration, conditions such as type II
diabetes are ongoing, and any treatment regime will typically
involve long-term, ongoing therapy, for periods of months or years.
Currently available GLP-1 therapies are typically injectables which
require administration around twice a day for the period of
treatment. This will generally be by patient self-administration.
Since frequent injection over a long period is not an optimal
administration strategy, there is clearly scope for GLP-1 users to
benefit from long-acting, sustained formulations, which might be
administered much less frequently.
[0024] The only long-acting GLP-1 product known to be in
development is Exenatide LAR, developed by a collaboration of
Alkermes, Amylin and Lilly. This uses the Alkermes Medisorb.RTM.
delivery system consisting of microspheres of biodegradable
polymers. The release system comprises a poly(DL-lactide) (PDLL)
polymer microsphere formulation suspended in water, which entraps
the GLP-1 analogue exenatide. The loading level of exenatide in
Exenatide LAR is typically 0.8 to 2 mg per dose administered
weekly. Evidently, since patients undergoing treatment with a GLP-1
receptor agonist will typically require ongoing treatment for many
months or years, a depot system allowing loading and controlled
release of a larger dose over a longer period would offer a
considerable advantage.
[0025] 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.
Evidently, it would be an advantage to provide a system of low
viscosity, such as a homogeneous solution, dispersion of fine
particles, or L2 phase, which could be administered easily through
a narrow needle, thus decreasing the discomfort of the patient
during the procedure. In the case of type II diabetes, this ease of
administration is particularly significant because most patients
will currently be on a self-administration regime. 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.
[0026] The poly-lactate, poly-glycolate and
poly-lactate-co-glycolate polymers typically used for degrading
slow-release formulations, and which are used in the only known
GLP-1 sustained release product, 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, the discomfort at the site of administration and
the formation of connective scar tissue are greater than
desirable.
[0027] 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.
[0028] Evidently, in the case of GLP-1 analogues, it is important
that the "burst" period, immediately after administration, is not
so pronounced that it causes hypoglycaemia in the subject. GLP-1 is
much safer in this respect than insulin, but clinical trials of
some GLP-1 analogues have shown hypoglycaemic effects with
non-sustained release formulations, and the dose injected when a
formulation is designed to last for several weeks will be
correspondingly higher. It would therefore be a considerable
advantage to minimise the immediate "burst" effect upon
administration of a GLP-1 analogue composition.
[0029] 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.
[0030] 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.
[0031] An alternative, lipid-based, slow-release composition of
GLP-1 and analogues thereof is described in WO2006/131730. This is
a highly effective formulation, but the concentration of GLP-1
analogue which can be included in the formulation is limited by the
solubility of the (peptide) active agent. Evidently, a higher
concentration of active agent allow 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 GLP-1 or GLP-1 analogues
could be included in a lipid-based depot formulation.
[0032] The present inventors have now established that by providing
a non-aqueous pre-formulation comprising a non-polymeric
slow-release vehicle, at least one peptide active agent (such as at
least one GLP-1 receptor agonist), at least one lipid soluble acid
and a biologically tolerable solvent in a low viscosity phase, such
as molecular solution, a pre-formulation may be generated
addressing many of the shortfalls of known depot formulations, and
which may be applied to provide a GLP-1 receptor agonist depot. In
particular, the pre-formulation is easy to manufacture, may be
sterile-filtered, has low viscosity (allowing easy and less painful
administration typically through a narrow needle), allows a higher
level of bioactive agent to be incorporated than has previously
been demonstrated (thus potentially allowing a smaller amount of
composition to be used), requires shallow injection and/or forms a
desired non-lamellar depot composition in vivo having a
controllable "burst" or "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. Evidently, these
advantages apply equally to other suitable active agents, and in
particular to peptides.
[0033] In a first aspect, the present invention thus provides a
non-aqueous pre-formulation comprising a low viscosity mixture
of:
[0034] i) a non-polymeric slow-release matrix
[0035] ii) at least one biocompatible, (preferably oxygen
containing) organic solvent;
[0036] iii) at least one peptide active agent; and
[0037] iv) at least one lipid soluble acid;
[0038] In one preferred aspect, the peptide active agent is a GLP-1
receptor agonist and the non-polymeric slow-release matrix is a
lipid based slow-release matrix. In a second aspect, the invention
therefore provides a non-aqueous pre-formulation comprising a low
viscosity mixture of:
[0039] i) a lipid-based slow-release matrix
[0040] ii) at least one biocompatible, (preferably oxygen
containing) organic solvent;
[0041] iii) at least one peptide active agent, such as a GLP-1
receptor agonist; and
[0042] iv) at least one lipid soluble acid;
[0043] In a further aspect, the invention provides a non-aqueous
pre-formulation comprising a low viscosity mixture of:
[0044] a) at least one neutral diacyl lipid and/or a
tocopherol;
[0045] b) at least one phospholipid;
[0046] c) at least one biocompatible, (preferably oxygen
containing) organic solvent;
[0047] d) at least one peptide active agent, such as a GLP-1
receptor agonist; and
[0048] e) at least one lipid soluble acid;
[0049] wherein the pre-formulation forms, or is capable of forming,
at least one liquid crystalline phase structure upon contact with
an aqueous fluid.
[0050] In one preferred embodiment, this non-aqueous
pre-formulation will comprise a low viscosity mixture of:
[0051] a) at least one diacyl glycerol;
[0052] b) at least one phosphatidyl choline;
[0053] c) at least one oxygen containing organic solvent;
[0054] d) at least one peptide active agent, such as a GLP-1
receptor agonist; and
[0055] e) at least one lipid soluble acid;
[0056] wherein the pre-formulation forms, or is capable of forming,
at least one liquid crystalline phase structure upon contact with
an aqueous fluid.
[0057] Generally, the aqueous fluid will be a body fluid,
particularly extra-vascular fluid, extracellular fluid/interstitial
fluid or plasma, and the pre-formulation will form a liquid
crystalline phase structure when contacted with such a fluid (e.g.
in vivo). The pre-formulation of the invention will generally not
contain any significant quantity of water prior to
administration.
[0058] In a further aspect of the invention, there is also provided
a method of delivery of a peptide active agent (especially a GLP-1
receptor agonist) to a human or non-human animal (preferably
mammalian) body, this method comprising parenterally administering
(e.g. i.m. or preferably s.c.) a non-aqueous pre-formulation
comprising a low viscosity mixture of:
[0059] i) a non-polymeric slow-release matrix
[0060] ii) at least one biocompatible, (preferably oxygen
containing) organic solvent;
[0061] iii) at least one peptide active agent (preferably a GLP-1
receptor agonist); and
[0062] iv) at least one lipid soluble acid;
[0063] In a preferred aspect, the non-aqueous pre-formulation
comprises a low viscosity mixture of as described in a preferred
aspect or embodiment described herein.
[0064] In a further aspect, the present invention also provides a
method for the preparation of a depot composition comprising
exposing a non-aqueous pre-formulation comprising a low viscosity
mixture of:
[0065] i) a non-polymeric slow-release matrix
[0066] ii) at least one biocompatible, (preferably oxygen
containing) organic solvent;
[0067] iii) at least one peptide active agent (preferably a GLP-1
receptor agonist); and
[0068] iv) at least one lipid soluble acid;
[0069] to an aqueous fluid in vivo.
[0070] Preferably the pre-formulation administered is a
pre-formulation of the present invention as described herein in any
of the aspects of the invention.
[0071] In a still further aspect the present invention provides a
process for the formation of a non-aqueous pre-formulation suitable
for the administration of a peptide bioactive agent to a
(preferably a human or non-human mammalian) subject, said process
comprising forming a low viscosity mixture of
[0072] i) a non-polymeric slow-release matrix
[0073] ii) at least one biocompatible, (preferably oxygen
containing) organic solvent;
[0074] and dissolving or dispersing at least one peptide active
agent (preferably a GLP-1 receptor agonist); and GLP-1 receptor
agonist and at least one lipid soluble acid in the low viscosity
mixture, or in at least one of components i) or ii) prior to
forming the low viscosity mixture. Preferably the non-aqueous
pre-formulation so-formed is a formulation of the invention as
described herein, and in particular, component i) preferably
comprises a lipid matrix as described herein, especially a lipid
matrix comprising components a) and b) as indicated herein.
Preferably, the lipid soluble acid component is added prior to
addition of the peptide active agent (e.g. GLP-1 receptor agonist)
component.
[0075] In a yet still further aspect the present invention provides
the use of a low viscosity mixture of:
[0076] i) a non-polymeric slow-release matrix
[0077] ii) at least one biocompatible, (preferably oxygen
containing) organic solvent;
[0078] iii) at least one peptide active agent (preferably a GLP-1
receptor agonist); and
[0079] iv) at least one lipid soluble acid;
[0080] in the manufacture of a non-aqueous pre-formulation for use
in the sustained administration of said peptide active agent (e.g.
GLP-1 receptor agonist). Preferably the pre-formulation is as
described in the preferred aspects of the present invention, and
may be, for example a low viscosity mixture of:
[0081] a) at least one neutral diacyl lipid and/or a
tocopherol;
[0082] b) at least one phospholipid;
[0083] c) at least one biocompatible, (preferably oxygen
containing) organic solvent;
[0084] d) at least one peptide active agent, such as a GLP-1
receptor agonist; and
[0085] e) at least one lipid soluble acid;
[0086] wherein said pre-formulation is capable of forming at least
one liquid crystalline phase structure upon contact with an aqueous
fluid.
[0087] The invention also provides the use of a pre-formulation, or
depot composition of the invention in therapy, and in the
manufacture of a medicament for treatment of an appropriate medical
indication. In particular, in one aspect, the invention provides
for the use of a GLP-1 receptor agonist-containing composition as
described herein in the manufacture of a medicament for the
treatment of diabetes, especially type II diabetes, or for the
medical or cosmetic treatment of excess bodyweight and/or obesity.
In the case of medical treatment, the composition is typically
administered to a subject in medical need thereof (e.g. having
diabetes, excess bodyweight or obesity). In the case of cosmetic
treatment, the subject may not have an identifiable medical need
thereof, but may, for example, have a body mass index in the
slightly overweight, higher normal, or normal range, wherein the
benefit from weight loss is largely or solely cosmetic rather than
medical.
[0088] In a still further aspect, the present invention provides a
method for the treatment of a human or non-human mammalian subject
comprising administering to said subject any of the non-aqueous
pre-formulations described herein. Preferably, in such an aspect,
the present invention provides a method for the treatment of a
human or non-human mammalian subject in need thereof with a GLP-1
receptor agonist, said method comprising administering to said
subject a GLP-1 receptor agonist-containing pre-formulation as
described herein, preferably a non-aqueous pre-formulation
comprising a low-viscosity mixture of;
[0089] a) at least one neutral diacyl lipid and/or a
tocopherol;
[0090] b) at least one phospholipid;
[0091] c) at least one biocompatible, (preferably oxygen
containing) organic solvent;
[0092] d) at least one GLP-1 receptor agonist; and
[0093] e) at least one lipid soluble acid;
[0094] Preferably, the method of treatment is a method for the
treatment of at least one condition selected from diabetes, type I
diabetes, type II diabetes, excess bodyweight and obesity.
Alternatively the method may be a method of cosmetic treatment
(e.g. to aid in the reduction of body weight) of a healthy subject
(e.g. one having a normal BMI). Such a method of cosmetic treatment
may exclude medical treatment, and thus be a method of cosmetic but
not medical treatment.
[0095] The invention further provides a method of treatment
comprising administration of a GLP-1 receptor agonist composition
as described herein, especially in a subject in need thereof. The
method of treatment is particularly for the treatment of diabetes,
especially type II diabetes.
[0096] In a yet further aspect, the present invention provides the
use of;
[0097] a) at least one neutral diacyl lipid and/or a
tocopherol;
[0098] b) at least one phospholipid;
[0099] c) at least one biocompatible, (preferably oxygen
containing) organic solvent;
[0100] d) at least one GLP-1 receptor agonist; and
[0101] e) at least one lipid soluble acid;
[0102] in the manufacture of a low viscosity pre-formulation
medicament for use in the in vivo formation of a depot for
treatment of type I diabetes, type II diabetes, excess bodyweight
and/or obesity.
[0103] In a still further aspect, the invention provides for the
use of at least one lipid soluble acid in increasing the use of a
lipid soluble acid in the stabilisation of at least one peptide
active agent (e.g. a GLP-1 receptor agonist) in a lipid-based
composition, the lipid based composition comprising a low viscosity
mixture of;
[0104] i) a non-polymeric slow-release matrix
[0105] ii) at least one biocompatible, (preferably oxygen
containing) organic solvent;
[0106] iii) said at least one peptide active agent; and
[0107] iv) said at least one lipid soluble acid;
[0108] In a yet still further aspect, the invention provides a
method of stabilising at least at least one peptide active agent
(e.g. a GLP-1 receptor agonist) in a lipid-based composition, said
method comprising formulating said at least one peptide active
agent with at least one lipid soluble acid in a the lipid
composition, as a low viscosity mixture of,
[0109] i) a non-polymeric slow-release matrix
[0110] ii) at least one biocompatible, (preferably oxygen
containing) organic solvent;
[0111] iii) said at least one peptide active agent; and
[0112] iv) said at least one lipid soluble acid;
[0113] The preferred non-polymeric slow-release matrices are those
indicated herein in respect of any aspect of the invention.
[0114] By "stabilising" is indicated an increase in solubility or
dispensability of a component (especially an active agent) in the
non-polymeric (e.g. lipid-based) matrix, 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 lipid soluble acid
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
lipid soluble acid. Equally, an increase in stability may be
demonstrated by the chemical and/or physical stability of a peptide
active agent in a matrix for a greater period than would be
observed in the absence of a lipid soluble acid. 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.
[0115] In all aspects of the present invention, the preferred
non-polymeric slow-release matrix component i) is preferably a
lipid-based or acyl-saccharide-based matrix, and in particular,
"non-polymeric" is used to indicate that the matrix does not
contain any significant quantity of poly-lactate, poly-glycolate or
poly-glycolate-co-lactate polymer (e.g. no more than 1% by
weight).
[0116] A preferred acyl-saccharide-based matrix is acylated
sucrose, particularly sucrose acetate isobutyrate, including the
fully-acylated sucrose ester having two acetate groups to six
isobutyrate groups sold by the DURECT corporation of California,
USA as "SABER". The matrix component may consist of at least 80%
acyl-saccharides and will preferably consist essentially of such in
this embodiment.
[0117] The most preferred non-polymeric slow-release matrix in all
aspects of the invention is a lipid-based matrix, which is to say a
matrix comprising at least 80% and preferably consisting
essentially of lipid (i.e. amphiphilic) components.
[0118] In the aspects of the invention relating to lipid-based
systems, the lipid matrix will preferably be components a) and b),
as indicated herein. Components a) and b) will preferably be:
[0119] a) at least one diacyl glycerol and/or at least one
tocopherol;
[0120] b) at least one phosphatidyl choline;
[0121] In all aspects, component c) will preferably be at least one
oxygen containing organic solvent.
[0122] 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.
[0123] 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 non-aqueous
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.
[0124] 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 (e.g. at least one GLP-1 receptor agonist), 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.
[0125] The invention provides for a pre-filled administration
device as indicated herein and a kit as indicated herein comprising
a non-aqueous pre-formulation as described herein.
[0126] 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 i) and ii) (e.g. components a) to
c)), as described here, and a second containing a measured dose of
at least one peptide active agent (e.g. at least one GLP-1 receptor
agonist) as described herein. The lipid soluble acid iv) (component
e)) may be formulated with the active agent, or more preferably as
part of the low viscosity mixture, which will then comprise
components i), ii) and iv) (e.g. a) to c) and e)).
[0127] Such a "two component kit" may comprise the peptide active
agent (e.g. GLP-1 analogue) as a powder formulation (optionally
including component iv)) in one vial or pre-filled syringe and the
matrix and solvent components of the formulation (i.e. components
i), and ii, with or without iv), e.g. components a) to c) (and
preferably e))) 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 peptide 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 i) 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.
[0128] In this aspect, the invention therefore provides a two
component kit comprising
[0129] i) a first vessel containing a low viscosity mixture of
components i) and ii) (preferably a) to c)) as described
herein;
[0130] ii) a second vessel containing at least one peptide active
agent (preferably at least one GLP-1 receptor agonist),
[0131] iii) a lipid soluble acid optionally in a third vessel,
preferably in the second vessel, or most preferably in the first
vessel;
[0132] iv) optionally and preferably at least one of: [0133] 1) at
least one syringe (which may be one or both of said first and
second vessels); [0134] 2) a needle for administration, such as
those described herein; [0135] 3) instructions for generation of a
composition of the invention from the contents of the first and
second vessels; [0136] 4) instructions for administration, whereby
to form a depot as described herein.
[0137] 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
matrix for use in the present invention is that 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.
[0138] 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 at 20.degree. C.
[0139] It has been observed that by the addition of small amounts
of low viscosity 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.
[0140] In one preferred aspect, the present invention provides a
pre-formulation comprising components a, b, c, e and at least one
GLP-1 receptor agonist as indicated herein. The amounts of these
components will typically be in the range 30-70% a), 30-60% b) and
0.1-20% c), with the GLP-1 receptor agonist present at 0.01% to
10%, (such as 40-70% a), 30-60% b) and 0.1-10% c), with the GLP-1
receptor agonist present at 0.1% to 10%).
[0141] Typically, component e) is present at a peptide active agent
(e.g. GLP-1 receptor agonist) to lipid soluble acid molar ratio of
1:1 to 1:30, preferably 1:1 to 1:20, and most preferably 1:1 to
1:15, e.g. 1:2 to 1:10. Since typical lipid soluble acid are of
lower molecular weight than peptides such as the GLP-1 receptor
agonist, the proportion by weight of lipid soluble acid may be
relatively small. For example, with a small molecular weight pH
adjuster (e.g. less than 500 amu), 0.1 to 5% of the composition may
be lipid soluble acid, preferably 0.2 to 2%.
[0142] All % are specified by weight herein throughout, unless
otherwise indicated. The formulations may consist of essentially
only these components and in one aspect consist entirely of such
components.
[0143] Preferable ranges for component a) are 33-60% (e.g. 43-60%),
particularly 35-55% (e.g. 40-55%) and preferable ranges of
component b) are 33-55% (e.g. 35-55%), particularly 35-50% (e.g. 40
to 50%).
[0144] Ratios of a:b are typically 40:60 to 70:30, preferably 45:55
to 60:40 and more preferably 48:52 to 55:45. Ratios of around 50:50
are highly effective.
[0145] The amount of solvent component ii) (e.g. component c)) in
the non-aqueous 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 20%, particularly 0.1 to 15% solvent will provide
suitable release and viscosity properties. This will preferably be
2 to 15% (e.g. 2 to 12%) and an amount of around 10% is highly
effective.
[0146] One advantage of the compositions of the present invention
over polymer formulations, such as PLGA spheres, is the 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 is 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), more
preferably less than 10% and most preferable less than 7%. This
applies particularly to the acyl saccharide and lipid aspects of
the invention and is discussed in more detail in WO
2005/117830.
[0147] 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 i) and ii) (a, b and c for the lipid aspect)
and will be easily determined for any particular combination of
components by standard methods.
[0148] Where a lipid matrix is used, the phase behaviour may be
analysed by techniques such as visual observation in combination
with polarized light microscopy, nuclear magnetic resonance, and
cryo-transmission electron microscopy (cryo-TEM) to look for
solutions, L2 or L3 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.
[0149] The preferred lipid-based matrix systems described herein
comprise lipid components a) and b), plus solvent (c), active agent
(d) and lipid-soluble acid (e) components. 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 caproyl (C6:0),
capryloyl (C8:0), capryl (C10:0), 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.
[0150] Mixtures of any number of diacyl lipids may be used as
component a. Preferably this component will include at least a
portion of glycerol dioleate (GDO). A highly preferred example is
DAG comprising at least 50%, preferably at least 80% and even
comprising substantially 100% GDO.
[0151] 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.
[0152] 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.
[0153] 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.
[0154] 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.
[0155] 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.
[0156] A particularly favoured combination of components a and b
are GDO with PC, especially GDO with soy PC. 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.
[0157] Component iii) (including component c) as appropriate) of
the pre-formulations of the invention is an organic solvent,
preferably an oxygen containing organic solvent. Since the
pre-formulation is to generate a depot composition following
administration (e.g. in vivo), typically upon contact with an
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.
[0158] In a preferred version, the solvent is such that a
relatively small addition to the matrix composition (e.g. 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% solvent can give a reduction
of two, or more orders of magnitude in viscosity over the
solvent-free composition, or over a matrix containing an unsuitable
solvent such as water, or glycerol.
[0159] Typical solvents suitable for use in the invention include
at least one solvent selected from alcohols, ketones, esters
(including lactones), ethers, amides and sulphoxides. Alcohols are
particularly suitable and form the preferred class of solvents.
Examples of suitable alcohols include ethanol, isopropanol, benzyl
alcohol and glycerol formal. Ethanol is most preferred. Monools are
preferred to diols and polyols. Where diols or polyols are used,
this is preferably in combination with an at least equal amount of
monool or other preferred solvent. Alternatively, diols such as
propylene glycol may be used alone or more preferably with at least
one fifth of the amount of mono-ol, especially ethanol (by weight).
Examples of ketones include acetone, and propylene carbonate.
Suitable ethers include diethylether, glycofurol, diethylene glycol
monoethyl ether, dimethylisobarbide, and polyethylene glycols.
Suitable esters include ethyl acetate, benzyl benzoate and
isopropyl acetate and dimethyl sulphide is as suitable sulphide
solvent. Suitable amides include n-methyl pyrrolidone (NMP),
2-pyrrolidone and dimethylacetamide (DMA). Sulphoxides include
methylsulphoxide and dimethylsulphoxide (DMSO).
[0160] A highly preferred combination for the lipid matrix aspect
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.
[0161] It is preferable that little or none of component c contains
halogen substituted hydrocarbons since these tend to have lower
biocompatibility. Where a portion of halogenated solvent such as
dichloromethane or chloroform is necessary, this proportion will
generally be minimised.
[0162] 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.
[0163] 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.
[0164] The pre-formulations of the present invention typically do
not contain significant amounts of water. Since it is essentially
impossible to remove every trace of water from a lipid composition,
this is to be taken as indicating that only such minimal trace of
water exists as cannot readily be removed. Such an amount will
generally be less than 1% by weight, preferably less that 0.5% by
the weight of the pre-formulation. In one preferred aspect, the
pre-formulations of the invention do not contain glycerol, ethylene
glycol or propylene glycol and contain no more than a trace of
water, as just described. The possible exception to this is when a
larger amount of water is balanced by an appropriate amount of
water-soluble organic solvent and is described above.
[0165] The pre-formulations of the present invention contain one or
more peptide active agents. Such 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.
[0166] Typical peptide actives will be in the range of 500 to
100,000 amu in molecular weight and evidently include protein
active agents. In one embodiment, the peptides 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 are
particularly preferred and therefore in one embodiment of the
invention, the active agent is a peptide acetate. In an alternative
embodiment, the peptide active agent may be essentially neutral,
and may have an isoelectric point of between pH5 and pH8,
preferably between pH5.2 and pH 7.5.
[0167] 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).
[0168] In one preferred embodiment of the present invention, the
peptide active agent will not be a somatostatin, or any analogue or
derivative thereof.
[0169] Most preferred active agents, which are used by way of
example throughout the present specification are GLP-1 receptor
agonists. Since GLP-1 is a peptide hormone, typical GLP-1 receptor
agonists will be the native GLP-1 and its analogues. Generally,
these will be peptides, especially of around 30 amino acids, e.g.
20 to 48, especially 25 to 45 (e.g. 25 to 38). Preferably such
peptides will be structurally related to GLP-1, extendin-4 and/or
one or more of the known analogues, including those listed here.
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., L-
or D-amino acids) and their analogues and derivatives. Preferred
amino acids include those listed above as constituents of the known
GLP-1 analogues.
[0170] 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).
[0171] By "GLP-1 receptor agonist", as used herein is indicated any
peptide agonist of the GLP-1 receptor, including all modified and
non-natural peptides such as those described herein, explicitly
including those described herein in the background section.
Preferably, these will be GLP-1 analogues or analogues of the
naturally occurring agonist "exendin-4", including naturally
occurring forms of GLP-1 and exendin-4, either human or from any
other species. These analogues are peptides, peptide derivatives or
peptide mimics. Peptide derived GLP-1 receptor agonists are most
preferred, such as those indicated above and especially
GLP-1(7-37), GLP-1(7-36) amide, Liraglutide, AVE-010 (ZP10),
TH0318, BIM 51077, NN2211, CJC-1131, LY315902 and Exenatide
(exendin-4). The specific sequences of several preferred GLP-1
receptor agonists are shown herein above, and these are all highly
suitable. Additional suitable GLP-1 receptor agonists are also
provided in the literature, and in particular these include the
peptides and acyl peptides listed in J Med Chem 43, 1664-1669
(2000) (especially the GLP-1 analogues shown in table 1 on page
1665); the derivatives made possible by the work provided in J Med
Chem. 47, 4128-4134 (2004), and in particular the structures
conforming to the essential amino acids, sites derivatisable with
fatty acids, and sites modifiable for improved peptidase resistance
as summarised in FIG. 3 on page 4130; the known analogues and all
analogues made available from the work described in Diabetes Metab.
Res. Rev. 21, 313-331 (2005), including the analogues described on
pages 322 to 323, plus analogues derivatised or formulated for
peptidase resistance as described on pages 323 to 325; and all
agonists described in Curr. Med. Chem. 10, 2471-2483 (2003), in
particular those with sequences and modifications described on page
2477. These documents, and in particular the parts indicated form
part of the disclosure of the present invention and are thus
explicitly incorporated herein by reference.
[0172] In one typical embodiment, the peptide active agent (e.g.
GLP-1 receptor agonist) 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% and more preferably 0.5 to 6%. These
levels may be applied to all aspects of the invention, where
context allows.
[0173] In a related embodiment, the peptide active agent (e.g.
GLP-1 receptor agonist) will be formulated at a level which cannot
easily be achieved in the absence of the lipid soluble acid
component of the mixture. In such an embodiment, the peptide active
agent (e.g. GLP-1 receptor agonist) content is typically at least
0.7%, preferably at least 1%, more preferably 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 GLP-1 receptor agonist), 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
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.
[0174] In one embodiment, the compositions of the present invention
rely upon the effect of the lipid soluble acid to allow for a
loading of peptide active agent (e.g. GLP-1 receptor agonist) at a
level above that which could be achieved in the absence of that
component. Obviously, a high loading level is highly advantageous
and is has been surprisingly established by the present inventors
that by including the lipid soluble acids specified herein, in the
amounts indicated, a much higher loading of peptide active agents
(particularly GLP-1 receptor agonists) can be obtained (see
examples). The level of peptide active agent (e.g. GLP-1 receptor
agonist) which could be loaded in a composition is easily
established by equilibration of the composition with excess active
agent (e.g. by slow end-over-end rotation for 5 days at 25.degree.
C.--see Examples). The present compositions can and preferably do
contain a greater amount of peptide active agent (particularly
GLP-1 receptor agonist) than can be achieved by equilibration in
the absence of the lipid soluble acid component. This can apply in
all aspects of the present invention, where context allow.
[0175] Where the peptide active agent is a GLP-1 receptor agonist,
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
P80 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 for a duration of 1 to 24
weeks, preferably 2 to 16 (e.g. 8, 10 or 12) weeks. 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, such as one of the GLP-1 receptor agonists indicated
herein. Evidently also, the biological half-life of the specific
active will be particularly important. The half-life of native
human GLP-1 (GLP-1(7-37) and GLP-1(7-36)amide), which is one
preferred active, 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 exenatide, 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.
[0176] It is a remarkable development of the present formulations
that very short half-life peptide active agents, including native
human GLP-1 (GLP-1(7-37) and GLP-1(7-36)amide) 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). Such a high performance in
delivery of a short half-life active is not otherwise known and no
other lipid depot system capable of sustained release of native
human GLP-1 has been reported. Thus, in one embodiment, the active
agent has a half-life of less than 1 hour, e.g. less than 15
minutes (such as GLP-1(7-37)) 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.
[0177] The "lipid soluble acid" component "e)" as used herein is
generally a low molecular weight compound which would form an
acidic solution in an aqueous medium (i.e. in water). Although
referred to as an "acid" herein, and acting as an acid in aqueous
solutions, this component does not generally act as a typical acid
in the pre-formulations of the invention, since these are
non-aqueous. In a preferred embodiment, such a lipid soluble acid
has a molecular weight of less than 500 amu, e.g. less than 300 amu
and more preferably less than 200 amu. Organic and mineral acids
form preferred lipid soluble acids, especially those having low
molecular weight as indicated. The lipid soluble acids will
generally be those having a pKa of lower than 5, preferably lower
than 4.7 and most preferably lower than 4.5. The acids must also be
suitable for dissolution at the required level in the chosen matrix
system. As the matrices are generally hydrophobic or amphiphilic,
suitable acids are referred to herein as "lipid soluble". The
suitability of any acid in any particular matrix system will be
established by one of ordinary skill by simple routine testing.
Since the lipid soluble acids are to be administered as part of a
parenteral drug-release system, biocompatibility in the relevant
quantities is also necessary. Particularly preferred lipid soluble
acids are selected from benzoic acid, citric acid, sulphonic acids
(e.g. methane sulphonic acid, benzene sulphonic acid or toluene
sulphonic acid) and hydrohalic acids (e.g. hydrochloric acid,
hydrobromic acid or hydoriodic acid). Most preferred lipid soluble
acids are benzoic acid, citric acid, methane sulphonic acid,
benzene sulphonic acid, toluene sulphonic acid and HCl.
[0178] In one alternative embodiment of the invention, the lipid
soluble acid is not a hydrohalic acid (e.g. not HCl, not HBr and/or
not HI). In this embodiment it is preferred that the lipid soluble
acid is benzoic acid, citric acid or a sulphonic acid.
[0179] The lipid soluble acids are referred to herein as "acids"
and in one preferred aspect they are formulated as at least
essentially consisting of the acid in free acid form. In an
alternative aspect, however, the lipid soluble acid may be the salt
of the corresponding acid as described herein, wherein the
counter-ion is a physiologically acceptable ion such as an
alkali-metal or alkaline earth metal cation, an ammonium ion or a
substituted ammonium ion. A mixture of such ions is evidently also
suitable. In one corresponding embodiment, the counter-ion is the
cation of the peptide active agent, or a mixture of ions including
the cation of the peptide active agent.
[0180] The function of the lipid soluble acids in the compositions
of the present invention is not immediately evident. Since the
compositions are essentially free of water, the aqueous hydrogen
ion concentration, which is the normal basis of pH, does not
directly apply, and the lipid soluble acids must have an additional
effect in these systems. Without being bound by theory, it is
believed that the ions of the lipid soluble acids serve to
stabilise the dissolution of the peptide(e.g. GLP-1 receptor
agonist) active agents, which are typically also formulated as
salts (generally the acetate salt). It has, however, been observed
by the present inventors that the free acid form of the
lipid-soluble acids is significantly more effective in stabilising
dissolution of high levels of active agent than the corresponding
salt. This is thought to be the result of the lower lipid
solubility of the ionic form, especially where the positive
counter-ion is poorly lipid-soluble. As a result, it is preferred
that the lipid soluble acids are used in their free-acid form, or
where a poly-acid is used, that all acid groups are in the
free-acid form. This may be the case in all embodiments of the
invention, but applies particularly to GLP-1 receptor agonists.
Where an acid salt is used, the counter-ion must evidently be
biotolerable, but it is preferable that this is has an organic
counter ion, such as an ammonium ion (e.g. R.sub.4N.sup.+ where the
four R groups are each H or C.sub.1 to C.sub.6 organic (e.g.
hydrocarbyl or heterocyclic) groups, which may joint to form rings,
and preferably no more than three R groups are H). The lipid
soluble acids preferably do not comprise metal ions, (e.g. alkali
metal or alkaline earth metal ions), such as sodium, potassium,
magnesium or calcium ions. Sodium ions of the lipid soluble acids
(e.g. sodium citrate) are preferably not present in the
formulations, and/or are not added thereto or formulated therewith.
Again, this may be the case in all embodiments of the invention,
but applies particularly to GLP-1 receptor agonists.
[0181] In all aspects of the invention, the lipid soluble acid
(component iv)/e)) is typically present at a molar ratio of peptide
active agent to lipid soluble acid of 1:1 to 1:30, preferably 1:1
to 1:20, e.g. 1:1 to 1:15 and most preferably 1:2 to 1:10. Since
typical lipid soluble acid are of lower molecular weight that the
peptide active agent, the proportion by weight of lipid soluble
acid may be relatively small. For example, with a small molecular
weight pH adjuster (e.g. less than 500 amu), 0.1 to 5% of the
composition may be lipid soluble acid, preferably 0.2 to 2%.
[0182] A sugar component may be present in the compositions of the
present invention, and this may also serve to increase the loading
and stability of the active agent. Preferred sugar components
include lactose, and more preferably sucrose or trehalose. Where
present, the sugar component may be present at 0.1 to 20%,
preferably 0.5 to 10% by weight, more preferably 1 to 5% by
weight.
[0183] Alternatively, there may be no sugar component present, for
example, there may be no sucrose, no lactose, and/or no trehalose
present in the composition. These may also be present at a low
level, such that the sugars (e.g. sucrose, lactose, and/or
trehalose) may each independently be present at 0 to 1%, preferably
0 to 0.5%.
[0184] Where the lipid soluble acid is citric acid or a citrate
salt, the stabilisation of GLP-1 is very effective. In one
embodiment, citric acid allows compositions of GLP-1 in the absence
of any sugar component. Thus, where GLP-1 and citrate are used, it
can be that there is no sugar component present, and in particular
no sucrose, lactose or trehalose present. Alternatively, where
present, the sugar component is at less than 1 wt %, preferably
less than 0.5 wt %.
[0185] In an alternative enbodiment, there may be a sugar (such as
sucrose, lactose or preferably trehalose), present in the
composition and in particular may be present at greater than 1%,
for example at 1.5 to 5%. In such compositions, it is preferred
that where the peptide active agent is a GLP-1 receptor agonist
(especially a human GLP-1), the level of peptide active agent will
be greater than 2% by weight, preferably greater than 2.5% by
weight, e.g. at least 3% by weight. Suitable ranges include 2 to
15% by weight, e.g. 2.5 to 12% by weight, or 3 to 10% by weight of
the total composition. Thus, where citrate, GLP-1 and a sugar, such
as sucrose, lactose and/ore trehalose, (at a level of at least 1%,
e.g. 1-3%) are present in the formulation, the GLP-1 will
preferably be formulated at greater than 2% by weight.
[0186] In one particularly preferred embodiment of the present
invention, the compositions (preformulations and resulting depots)
may include at least one biocompatible polyethyleneoxide or
poly(ethylene glycol) (PEG) fragmentation agent, such as a PEG
grafted lipid and/or surfactant. These agents are useful in all
compositions, and are believed to increase the stability of the
peptide active agent (e.g. GLP-1 receptor agonist), even at low
concentrations. In a particularly advantageous embodiment, however,
they may be highly useful for providing lipid depots with shorter
duration (e.g. 5 to 30 days, especially 7 to 21 days). This is
because such a component will tend to fragment the depot into
smaller pieces in situ and thus the degradation of the depot will
not only be biodegradation but also "physical" erosion, thus
enabling faster release (but still without any significant burst).
These are most preferable with the lipid matrices described
herein.
[0187] If included in the lipid-based pre-formulation, the content
of such a fragmentation agent component, would be 0.1-30%, more
preferably 0.5-25% and most preferably 2-20%. In particular, 0.1 to
1% (preferably 0.2 to 0.7%) is particularly useful for stabilising
the active agent, such as GLP-1 receptor agonist, and 1 to 25%,
preferably 5 to 20% is beneficial in controlling the depot release
period. Another advantage of including a fragmentation agent is
that it may be beneficial from a chronic use point of view. Users
of GLP-1 receptor agonist depot products, as well as users of many
other peptide depot productsare typically long-term users, and such
a depot erodes faster and thus the depot will vanish quicker from
the injection site, allowing earlier re-use of the site and causing
a lesser build-up of connective tissue around the sites of
injection. Furthermore, the inclusion of such an agent may even
improve the already good biotolerability/biocompatibility.
[0188] The most preferred fragmentation agent is Polysorbate 80
(P80). Other useful agents include 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.
[0189] The peptide active (e.g. GLP-1 receptor agonist) 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),
small amounts of surface active agents (e.g. P80--see above),
antioxidants (such as ascorbic acid, EDTA and citric acid), amino
acids (such as methionine, glutamate, lysine etc.) and anionic
lipids and surface active agents (such as dioleoyl phosphatidyl
glycerol (DOPG), palmitoyloleoyl phosphatidylglycerol (POPG) and
oleic acid (OA)).
[0190] One preferred additive agent is a thiol-based antioxidant.
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.
[0191] Unfortunately, many common antioxidants are not highly
compatible with lipid systems. Indeed, the present inventors have
surprisingly 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.
[0192] 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
particularly class of antioxidants is unusually well suited for use
in these systems.
[0193] The present inventors have now established that thiolated
antioxidants, particularly mono-thioglycerol (MTG), cysteine, and
cysteine analogues such as N-acetyl cysteine, are highly effective
in lipid based systems, especially in combination with the lipid
soluble acids as indicated herein. Thus in a preferred embodiment
of the present invention, an antioxidant component is included
comprising a thiolated antioxidant, preferably thiolated sugar,
thiolated amino acid, a thiolated amino ester, or a thiolated
polyol. Mono-thioglycerol, N-acetyl cysteine or cysteine are
preferred thiolated antioxidants.
[0194] The antioxidant component is generally included in the range
0.01 to 2.0% by weight of the total pre-formulation. This is most
preferably 0.05 to 1.0%, and around 0.2 to 0.5% of antioxidant
(particularly MTG) is particularly preferred, especially in
combination with the other preferred components and ranges
indicated herein above and below.
[0195] The reason for the utility of thiolated antioxidants in
general and MTG in particular is not known. Without being bound by
theory, it is believed that MTG acts as an effective chain-breaking
donating antioxidant according to established mechanisms whereby
peroxyl radicals (ROO.) are neutralized. Their quenching by the
thiolated antioxidant breaks the cycle of further oxidative
degradation. Thiols such as MTG and N-acetyl cysteine may also
regenerate certain components from their oxidized forms.
[0196] Stability data using a number of different antioxidants
demonstrate that thiolated antioxidants are surprisingly more
efficient than other antioxidants in suppressing the oxidative
degradation of bioactive agents. Thiolated antioxidants can also
show a synergistic effect in combination with the lipid soluble
acids of the present invention, in maintaining the chemical and
physical stability of the peptide active agent and complete
pre-formulation.
[0197] 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.
[0198] 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.
[0199] The preferred lipid matrix-based 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 equality (e.g. around 35:65 to 65:35,
preferably 42:58 to 58:42, most preferably 46:54 to 54:46).
[0200] 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 spray 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 10 wt % of solvent (component c) having a
viscosity reducing effect. This is in contrast to a "concentrated"
or "unswollen" L.sub.2 phase containing no solvent, or a lesser
amount of solvent, or containing a solvent (or mixture) which does
not provide the decrease in viscosity associated with the
oxygen-containing, low viscosity solvents specified herein.
[0201] 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.
[0202] Without being bound by theory, it is believed that upon
exposure (e.g. to body fluids), 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). In the case of an
acyl-saccharide matrix, this with cause a rapid increase in
viscosity as solvent is lost, and in the case of a lipid matrix, 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).
[0203] 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. Comparative
tests have been carried out between the known PLGA slow-release
product and preferred formulations of the present invention
containing GDO, soy PC, ethanol and active agents. These indicate
that formulations of the present invention give lesser degradation
under simulated in vivo conditions than known compositions. The
formulations of the invention thus may provide in vivo depots of
GLP-1 receptor agonists which require administration only once
every 7 to 360 days (e.g. 20 to 360 days), preferably 30 to 240
days (e.g. 30 to 168 days), more preferably 60 to 180 days (e.g.
around 90 days, such as 60 to 120 or 90.+-.7 days). Alternatively,
in a further preferred embodiment, the durations are somewhat
shorter, preferably 10 to 240 days (e.g. 20 to 168 days), more
preferably 14 to 180 days (e.g. around 60 days, such as 6 to 10
weeks). 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.
[0204] 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 GLP-1 receptor
agonist 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.
[0205] 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.
[0206] 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:
[0207] All proportions indicated herein may optionally be varied by
up to 10% of the amount specified, optionally and preferably by up
to 5%;
[0208] Component a) comprises, consists essentially of or
preferably consists of GDO;
[0209] Component b) comprises, consists essentially of or
preferably consists of soy PC;
[0210] Component c) comprises, consists essentially of or
preferably consists of a 1, 2, 3 or 4 carbon alcohol, preferably
isopropanol or more preferably ethanol;
[0211] Component e) comprises, consists essentially of or
preferably consists of a sulphonic acid or hydrohalic acid,
preferably methane sulfonic acid (MeSulf), benzene sulfonic acid
(BzSulf), toluene sulphonic acid (TSulf), benzoic acid, citric acid
or anhydrous hydrogen chloride;
[0212] The pre-formulation contains at least one GLP-1 receptor
agonist selected from those described or referred to herein,
preferably GLP-1(7-37), GLP-1(7-36)amide, Liraglutide, AVE-010,
TH-0318, LY548806 or exenatide;
[0213] The pre-formulation has a low viscosity as indicated
herein.
[0214] The pre-formulation comprises a lipid matrix and forms a
liquid crystalline phase as indicated herein upon in vivo
administration.
[0215] The pre-formulation generates a depot following in vivo
administration, which depot releases at least one GLP-1 receptor
agonist at a therapeutic level over a period of at least 7 days,
preferably at least 21 days, more preferably at least 30 days.
[0216] The pre-formulation has a higher loading of peptide active
agent (e.g. GLP-1 receptor agonist) than is stable in the same
formulation in the absence of the pH adjusting (lipid soluble acid)
component e).
[0217] The pre-formulation has a higher loading of peptide active
agent (e.g. GLP-1 receptor agonist) than is obtainable by
equilibration at 25.degree. C. of the same formulation in the
absence of the pH adjusting component e).
[0218] 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:
[0219] The method comprises the administration of at least one
formulation with one or more preferred features as indicated
above;
[0220] The method comprises the administration of at least one
formulation as indicated herein by i.m., s.c. or preferably deep
s.c. injection;
[0221] The method comprises administration by means of a pre-filled
administration device as indicated herein;
[0222] The method comprises administration through a needle no
larger than 20 gauge, preferably smaller than 20 gauge, and most
preferably 23 gauge or smaller;
[0223] The method comprises a single administration every 7 to 360
days, preferably 7 to 120 days, more preferably 14 to 60 days.
[0224] The method comprises a single administration every 14 to 180
days, preferably around 60 days.
[0225] 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:
[0226] The use comprises the use of at least one formulation with
one or more preferred features as indicated above;
[0227] 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;
[0228] The use comprises the manufacture of a medicament for
administration by means of a pre-filled administration device as
indicated herein;
[0229] 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;
[0230] The use comprises the manufacture of a medicament for
administration once every 7 to 360 days, preferably 7 to 120 days,
more preferably 14 to 60 days.
[0231] 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:
[0232] They contain a preferred formulation as indicated
herein;
[0233] They comprise a needle smaller than 20 gauge, preferably no
larger than 23 gauge;
[0234] They contain a single dose of 0.05 to 250 mg of GLP-1
receptor agonist, preferably 0.1 to 100 mg and more preferably 1-50
mg;
[0235] They contain GLP-1(7-37), GLP-1(7-36)amide, TH-0318,
Liraglutide, exenatide or AVE-010, at around 0.05 to 250 mg;
[0236] They contain a homogeneous mixture of a composition of the
invention in ready-to-inject form.
[0237] They contain a formulation of components a) to c) for
combination with a GLP-1 receptor agonist whereby to form a
preformulation of the invention.
[0238] They contain a GLP-1 receptor agonist for combination with a
formulation of components a) to c), whereby to form a
preformulation of the invention.
[0239] They contain a total volume for administration of no more
than 5 ml, preferably no more than 3 ml more preferably no more
than 2 ml.
[0240] 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:
[0241] They contain a preferred formulation as indicated
herein;
[0242] They contain a pre-filled device as indicated herein;
[0243] They contain a needle smaller than 20 gauge, preferably no
larger than 23 gauge;
[0244] They contain a single dose of 0.05 to 250 mg of GLP-1
receptor agonist, preferably 0.1 to 100 mg and more preferably 1-50
mg;
[0245] They contain GLP-1(7-37), GLP-1(7-36)amide, TH-0318,
Liraglutide or AVE-010, at around 0.05 to 250 mg;
[0246] They contain a "two compartment kit" comprising at least two
vessels containing a lipid formulation of the invention and a GLP-1
receptor agonist powder, respectively.
[0247] They contain a total volume for administration of no more
than 5 ml, preferably no more than 3 ml more preferably no more
than 2 ml.
[0248] They contain instructions for administration by a route
and/or at a frequency as indicated herein;
[0249] They contain instructions for administration for use in a
method of treatment as described herein.
[0250] The Invention will now be further illustrated by reference
to the following non-limiting Examples and the attached Figures, in
which;
[0251] FIG. 1 shows the viscosity reducing effect on addition of
solvents.
[0252] FIG. 2 displays stability data supporting the highly
favourable storage stability obtained with formulation compositions
of the invention.
[0253] FIG. 3 shows the level of breakdown products in a
composition of the invention stored for four weeks at 5.degree.
C.
EXAMPLES
Example 1
Availability of Various Liquid Crystalline phases in the Depot by
Choice of Composition
[0254] Injectable formulations containing different proportions of
phosphatidyl choline (PC) ("SPC"--Lipoid S100) and glycerol
dioleate (GDO) and with ethanol (EtOH) as solvent were prepared to
illustrate that various liquid crystalline phases can be accessed
after equilibrating the depot precursor formulation with excess
water.
[0255] Appropriate amounts of PC, GDO and EtOH were weighed in
glass vials and the mixture was placed on a shaker until the PC
completely dissolved to form a clear liquid solution. GDO was then
added to form an injectable homogenous solution.
[0256] Each formulation was injected in a vial and equilibrated
with excess water. The phase behaviour was evaluated visually and
between crossed polarizes at 25.degree. C. Results are presented in
Table 1.
TABLE-US-00001 TABLE 1 Formulation PC (wt %) GDO (wt %) EtOH (wt %)
Phase in H.sub.2O A 22.5 67.5 10.0 L.sub.2 B 28.8 61.2 10.0 I.sub.2
C 45.0 45.0 10.0 I.sub.2/H.sub.II D 63.0 27.0 10.0
H.sub.II/L.sub..alpha. L.sub.2 = reversed micellar phase I.sub.2 =
reversed cubic liquid crystalline phase H.sub.II = reversed
hexagonal liquid crystalline phase L.sub..alpha. = lamellar
phase
Example 2
Viscosity in PC/GDO (5:5) or PC/GDO (4:6) on Addition of Solvent
(EtOH, PG and NMP)
[0257] A mixture of PC/GDO/EtOH with approximately 25% EtOH was
manufactured according to the method in Example 1. All, or nearly
all, of the EtOH was removed from the mixture with a rotary
evaporator (vacuum, 40.degree. C. for 1 h followed by 50.degree. C.
for 2 h) and the resulting mixture was weighed in a glass vial
after which 1, 3, 5, 10 or 20% of a solvent (EtOH, propylene glycol
(PG) or n-methyl pyrrolidone (NMP)) was added. The samples were
allowed to equilibrate several days before the viscosity was
measured with a CarriMed CSL 100 rheometer equipped with automatic
gap setting.
[0258] This example clearly illustrates the need for solvent with
certain depot precursors in order to obtain an injectable
formulation (see FIG. 1). The viscosity of solvent-free PC/GDO
mixtures increases with increasing ratio of PC. Systems with low
PC/GDO ratio (more GDO) are injectable with a lower concentration
of solvent.
Example 3
Degradation of Depot Formulation in the Rat
[0259] Various volumes (1, 2, 6 ml/kg) of the depot precursor (36%
wt PC, 54% wt GDO, and 10% wt EtOH) were injected in the rat and
were removed again after a period of 14 days. It was found that
substantial amounts of the formulations were still present
subcutaneously in the rat after this time, see Table 3.
TABLE-US-00002 TABLE 3 Dose (ml/kg) Mean diameter day 3 (mm) Mean
diameter day 14 (mm) 1 (n = 3) 15.8 12.5 2 (n = 3) 18.5 15.3 6 (n =
3) 23.3 19.3
Example 4
Preparation of a GLP-1 Formulation without pH-Adjusting Agent
[0260] The GLP-1 substance and the excipients used in example 4 to
8 are presented in the Table below.
TABLE-US-00003 GLP-1 substance and excipients used in the Examples
4 to 8: Name Abbreviation Supplier GLP-1(7-36)amide, acetate
GLP-1(Ac) PolyPeptide Laboratories, salt Inc., CA, USA
Phosphatidylcholine, soy SPC Lipoid, Germany Glycerol dioleate GDO
Danisco, Denmark Ethanol (99.5%) EtOH Kemetyl, Sweden Propylene
glycol PG Apoteket, Sweden
[0261] A lipid formulation comprising 1.08 g SPC, 1.08 g GDO, 0.08
g EtOH and 0.26 g PG was mixed in a 5 mL glass vial (composition:
SPC/GDO/EtOH/PG=43.2/43.2/3.2/10.4 wt %). The vial was placed on a
mixing table (end-over-end mixing) for approximately 2 hours at RT.
A transparent and homogenous formulation was obtained.
[0262] 0.02 g of GLP-1(Ac) was weighed into a 2 mL glass vial and
1.98 g of the lipid formulation, prepared as above, was added
(giving a total GLP-1(Ac) load of 1 wt %). The formulation was
mixed on a vortex mixer (to disperse the GLP-1(Ac) powder in the
formulation) and then placed on a mixing table at room temperature
for constant end-over-end mixing. After 5 days the sample still
contained a lot of undissolved GLP-1(Ac) as assessed visually and
the sample was therefore centrifuged at 5000 rpm for 15 minutes to
obtain a clear supernatant.
[0263] The GLP-1 concentration in the supernatant was assayed by a
normal phase (NP) HPLC method using UV detection.
[0264] The assayed GLP-1 (equivalents base--GLP-1(0)) concentration
in the sample was 6.25 mg/g (0.625 wt %). Because of the long
equilibration time used (5 days) this value is taken as the maximum
GLP-1 concentration achievable in the lipid formulation without the
addition of pH-adjusting agent.
Example 5
Preparation of GLP-1 Formulations with Methane Sulfonic Acid
(MeSulf) as pH-Adjusting Agent
[0265] A lipid formulation comprising SPC, GDO, EtOH, PG and MeSulf
(Sigma-Aldrich, Sweden) was prepared as described in Example 4. The
lipid composition was the following:
SPC/GDO/EtOH/PG/MeSulf=43.2/43.2/3.0/10.0/0.5 wt %.
[0266] The required amount of GLP-1(Ac) powder was weighed into 2
mL glass vials followed by addition of the lipid formulation in an
amount appropriate for achieving nominal drug loads of
approximately 3 to 6 wt % GLP-1(0). The samples were briefly
vortexed followed by continuous end-over-end rotation at room
temperature until completely homogenous and transparent samples
were obtained (1-3 days).
[0267] The concentration of GLP-1 (expressed as equivalents GLP-1
base =GLP-1(0)) in the respective formulations as determined by
HPLC is given in the Table below.
TABLE-US-00004 GLP-1 drug load in lipid formulations containing
MeSulf Assayed Excess Nominal (HPLC) GLP-1 load GLP-1(0) GLP-1(0)
compared with Formulation conc./wt % conc./wt % Example 4* A 3.00
3.07 4.91 B 3.73 3.77 6.03 C 4.61 4.58 7.33 D 5.43 5.36 8.54
*Calculated as the ratio between the assayed GLP-1(0) concentration
with MeSulf as pH-adjusting agent and the concentration found in
the formulation without MeSulf (Example 4)
Example 6
Preparation of GLP-1 Formulation with Anhydrous Hydrogen Chloride
(HCl) as pH-Adjusting Agent
[0268] A lipid formulation comprising SPC, GDO, PG and EtOH.HCl
(1.25M HCl in EtOH from Fluka, Sweden) was prepared as described in
Example 4. The lipid composition was the following:
SPC/GDO/PG/EtOH.HCl=43.0/43.0/10.0/4.0 wt %.
[0269] The required amount of GLP-1(Ac) powder was weighed into a 2
mL glass vial followed by addition of the lipid formulation. The
sample was briefly vortexed followed by continuous end-over-end
rotation at room temperature until a completely homogenous and
transparent sample was obtained (1 day).
[0270] The concentration of GLP-1 (expressed as GLP-1
base=GLP-1(0)) in the formulation as determined by HPLC is given in
the Table below.
TABLE-US-00005 GLP-1 drug load in lipid formulations containing HCl
Assayed Excess Nominal (HPLC) GLP-1 load GLP-1(0) GLP-1(0) compared
with Formulation conc./wt % conc./wt % Example 4* E 3.68 3.66 5.86
*Calculated as the ratio between the assayed GLP-1(0) concentration
with HCl as pH-adjusting agent and the concentration found in the
formulation without HCl (Example 4)
Example 7
Preparation of GLP-1 Formulations Containing Polysorbate 80 (P80)
and with Methane Sulfonic Acid (MeSulf) as pH-Adjusting Agent
[0271] A lipid formulation comprising SPC, GDO, P80 (Croda, USA),
EtOH, PG and MeSulf was prepared as described in Example 4. The
lipid composition was the following:
SPC/GDO/P80/EtOH/PG/MeSulf=41.0/41.0/5.0/3.0/10.0/0.5 wt %.
[0272] The required amount of GLP-1(Ac) powder was weighed into 2
mL glass vials followed by addition of the lipid formulation in an
amount appropriate for achieving nominal drug loads of 3, 4, 5 and
6 wt % GLP-1(0). The samples were briefly vortexed followed by
continuous end-over-end rotation at room temperature until
completely homogenous and transparent samples were obtained (1-3
days) indicating complete dissolution of GLP-1 in the lipid
formulation.
Example 8
Stability of GLP-1 in Formulations Containing MeSulf as
pH-Adjusting Agent
[0273] Lipid formulations containing MeSulf were prepared as
described in Example 4.
[0274] The required amount of GLP-1(Ac) powder was weighed into 6
mL glass vials followed by addition of the lipid formulation in an
amount appropriate for achieving nominal drug loads of 3 wt %
GLP-1(0). The samples were briefly vortexed followed by continuous
end-over-end rotation at RT until completely homogenous and
transparent samples were obtained (1-3 days). The nominal
composition of the samples is given in the Table below.
TABLE-US-00006 Nominal composition (wt %) of formulations for
stability study GLP- Formulation 1(0)* SPC DOPC** GDO PG EtOH
MeSulf F 3.0 41.7 -- 41.7 10.0 3.0 0.5 G 3.0 -- 41.7 41.7 10.0 3.0
0.5 *equivalents GLP-1 free base (GLP-1(0)). **Synthetic Dioleoyl
Phosphatidylcholine (DOPC) from Lipoid, Germany.
[0275] The samples were filled in 1 mL glass vials, capped with
Teflon-coated rubber stoppers and stored at 5.degree. C. in a
Termak climate chamber. After 4 weeks, the samples were taken out
for analysis of GLP-1 content, ID and degradation products using a
normal phase HPLC assay and UV detection (214 nm). The results
displayed in FIGS. 2 and 3 reveal highly favourable storage
stability of the formulations with essentially no degradation
(within the error limits of the assay) of GLP-1 during the
investigated time period.
* * * * *