(Table & Formula Not Copy)

FORM 2
THE PATENTS ACT, 1970
As amended by the Patents (Amendment) Act, 2002
and
The Patents Rules, 2003
As amended by the Patents (Amendment) Rules 2016
COMPLETE SPECIFICATION
(Section 10 and Rule 13)
TITLE OF THE INVENTION
GLUCAGON-LIKE PEPTIDE 1 (GLP-1) RECEPTOR AGONIST
COMPOSITIONS
APPLICANT
ARECOR LIMITED, a British company, having its address at Chesterford
Research Park, Little Chesterford, Saffron Walden CB10 1XL, United
Kingdom
PREAMBLE TO THE DESCRIPTION
The following specification particularly describes the invention and the manner in
which it is to be performed
2
This invention relates to aqueous solution compositions of glucagon-like peptide1 (GLP-1) receptor agonists and their use in therapy.
Background
5 Glucagon-like peptide-1 (GLP-1) is an incretin derived from the transcription
product of the proglucagon gene. The biologically active forms of GLP-1 are:
GLP-1(7-37) and GLP-1(7-36)NH2, fragments resulting from selective cleavage
of the proglucagon molecule.
10 GLP-1 (in its various biologically active forms) is an agonist of the glucagon-like
peptide 1 receptor (GLP-1 receptor) and has potent insulinotropic effects in
stimulating insulin secretion and inhibiting glucagon secretion. These glucoselowering effects make GLP-1 an important molecule in the treatment of diabetes.
One of the potential advantages of the use of GLP-1 over insulin therapy is that
15 insulin production is only stimulated when blood glucose levels are elevated,
thereby reducing the risk of hypoglycaemia.
GLP-1 is not used as a therapeutic agent directly because once in circulation it
has a half-life of less than 2 minutes, due to rapid degradation by the enzyme
20 dipeptidyl peptidase-4 (DPP4). However, a number of alternative GLP-1 receptor
agonists have been developed with an increased circulating half-life.
GLP-1 receptor agonists approved by regulatory authorities include:
• albiglutide (marketed under the tradenames Eperzan/Tanzeum)
25 • dulaglutide (marketed under the tradename Trulicity)
• exenatide (marketed under the tradenames Byetta/Bydureon)
• liraglutide (marketed under the tradenames Victoza/Saxenda)
• lixisenatide (marketed under the tradename Lyxumia/Adlyxin)
30 Tanzeum Pen for injection (for subcutaneous use) contains 67 mg lyophilized
albiglutide and 0.65 mL water for injection designed to deliver a dose of 50 mg in
a volume of 0.5 mL after reconstitution. Inactive ingredients include 153 mM
mannitol, 0.01% (w/w) polysorbate 80, 10 mM sodium phosphate, and 117 mM
trehalose dihydrate.
35
Trulicity is a clear, colourless, sterile solution. Each 0.5 mL of Trulicity solution
contains 0.75 mg or 1.5 mg of dulaglutide. Each single-dose pen or prefilled
syringe contains 0.5 mL of solution and the following excipients: citric acid
anhydrous (0.07 mg), mannitol (23.2 mg), polysorbate 80 (0.10 mg) and trisodium
40 citrate dihydrate (1.37 mg) in water for injection.
Byetta (exenatide injection) is supplied for subcutaneous injection as a sterile,
preserved isotonic solution in a glass cartridge that has been assembled in a
pen-injector. Each 1 mL contains 250 µg synthetic exenatide, 2.2 mg m-cresol as
3
an antimicrobial preservative, mannitol as a tonicity-adjusting agent, and glacial
acetic acid and sodium acetate trihydrate in water for injection as a buffering
solution at pH 4.5.
5 Bydureon is a white to off-white powder that is available in a dosage strength of 2
mg exenatide per vial or per pen. Exenatide is incorporated in an extendedrelease microsphere formulation containing a 50:50 poly(D,L-lactide-co-glycolide)
polymer (37.2 mg per dose) along with sucrose (0.8 mg per dose). The powder
must be suspended in the diluent prior to injection. The diluent is composed of
10 sodium carboxymethylcellulose (19 mg), polysorbate 20 (0.63 mg), sodium
phosphate monobasic monohydrate (0.61 mg), sodium phosphate dibasic
heptahydrate (0.51 mg), sodium chloride (4.1 mg) and water for injection.
Victoza is a clear, colourless or almost colourless solution. Each 1 mL of Victoza
15 solution contains 6 mg of liraglutide and the following inactive ingredients:
disodium phosphate dihydrate (1.42 mg); propylene glycol (14 mg); phenol (5.5
mg); and water for injection.
Saxenda is a clear, colourless solution. Each 1 mL of Saxenda solution contains
20 6 mg of liraglutide and the following inactive ingredients: disodium phosphate
dihydrate (1.42 mg); propylene glycol (14 mg); phenol (5.5 mg); and water for
injection. Each pre-filled pen contains a 3 mL solution of Saxenda equivalent to
18 mg liraglutide (free-base, anhydrous).
25 Lyxumia/Adlyxin injection is a sterile, clear, colourless aqueous solution for
subcutaneous administration supplied in two single-patient use prefilled pens.
Each prefilled pen contains 3 mL solution and each mL contains 50 or 100 µg
lixisenatide. Inactive ingredients are glycerol 85% (54 mg), sodium acetate
trihydrate (10.5 mg), methionine (9.0 mg), m-cresol (8.1 mg), and water for
30 injection.
The GLP-1 receptor agonist products are presented in pre-filled devices (typically
pre-filled pens) and are self-administered by the patients. However, in order to
maintain the stability of the GLP-1 receptor agonist in the composition during
35 storage and during the in-use period (i.e. the period following the initial use of the
pre-filled device during which the pre-filled pen is used repeatedly by the patient
until the pen cartridge becomes empty) patients must ensure that the devices are
stored and used correctly.
40 All of the currently marketed GLP-1 receptor agonist products have to be stored
at 2-8 °C for their entire shelf-life except the in-use period. During the in-use
period, after the initial use of the Victoza/Saxenda (liraglutide) pen, the pen can
be stored at temperatures up to 30 °C for up to 30 days. Trulicity (dulaglutide)
and Lyxumia/Adlyxin (lixisenatide) pens can be stored at temperatures up to 30
4
°C for up to 14 days after the initial use. Byetta (exenatide) pen can be stored at
temperatures up to 25 °C for up to 30 days. Bydureon (exenatide) vial can be
stored at temperatures up to 30 °C for up to 4 weeks prior its use, but must be
used immediately after reconstitution. Eperzan/Tanzeum (albiglutide) pens can
5 be stored at temperatures up to 30 °C for up to 4 weeks during the in-use period.
In order to improve patients’ convenience and shipment logistics there is a need
to improve the stability of compositions of GLP-1 receptor agonists. Desirable
compositions would have one or more of the following properties:
• the temperature at which the product can be kept during the in-use period
10 is increased, e.g. to 40 °C
• the duration of the in-use period is increased, e.g. to 1 month, to 2
months, and preferably to 3 months
• the product can be stored at increased temperature, such as controlled
room temperature (20-25 °C) for part of the shelf-life, e.g. 3 months, 6
15 months or the entire shelf-life, whilst maintaining the in-use stability at 30
°C.
Thus, an object of the present invention is the provision of an aqueous solution
composition of a GLP-1 receptor agonist as active ingredient with improved
20 stability.
Summary of the invention
According to the invention, there is provided an aqueous solution composition
25 comprising a GLP-1 receptor agonist as an active ingredient and multivalent
anions having a charge of at least minus 2 as stabilising agent, wherein the total
concentration of multivalent anions in the composition having a charge of at least
minus 2 is at least 15 mM.
30 Figures
Fig. 1: Stability of liraglutide in the presence of propylene glycol, mannitol,
trehalose or sodium chloride as additives, assessed by RP-HPLC following
storage at 30 °C for 9 and 14 weeks (see Table 1 of Example 1).
35
Fig. 2: Stability of liraglutide in the presence of propylene glycol, mannitol or
histidine (with and without mannitol) as additives, assessed by RP-HPLC
following storage at 30 °C for 9 and 14 weeks (see Table 1 of Example 1).
40 Fig. 3: Stability of liraglutide in the presence of citrate anions (with and without
mannitol), assessed by RP-HPLC following storage at 30 °C for 9 and 14 weeks
(see Table 2 of Example 2).
5
Fig. 4: Stability of liraglutide in the presence of phosphate anions (with and
without mannitol) assessed by RP-HPLC following storage at 30 °C for 9 and 14
weeks (see Table 2 of Example 2).
5 Fig. 5: Stability of liraglutide in the presence of sulphate anions (with or without
mannitol) assessed by RP-HPLC following storage at 30 °C for 9 and 14 weeks
(see Table 2 of Example 2).
Description of the sequence listings
10 SEQ ID NO. 1: GLP-1(7-37)
SEQ ID NO. 2: GLP-1(7-36)NH2
SEQ ID NO. 3: exendin-4/Exenatide
15
SEQ ID NO. 4: exendin-3
SEQ ID NO. 5: Albiglutide
20 SEQ ID NO. 6: Dulaglutide
SEQ ID NO. 7: Liraglutide
SEQ ID NO. 8: Lixisenatide
25
SEQ ID NO. 9: artificial sequence
SEQ ID NO. 10: artificial sequence
30 SEQ ID NO. 11: artificial sequence
SEQ ID NO. 12: exemplary serum albumin sequence
SEQ ID NO. 13: exemplary FcIgG4 sequence
35
Detailed description of the invention
As used herein a GLP-1 receptor agonist is any insulinotropic peptide which fully
or partially activates the human GLP-1 receptor. In one embodiment, the GLP-1
receptor agonist is any peptide that binds to a GLP-1 receptor with an affinity
40 constant (KD) of below 1 µM, for example below 100 nM as measured by
methods known in the art (see for example WO98/08871, the contents of which is
herein incorporated by reference) and exhibits insulinotropic activity, where
insulinotropic activity may be measured in vivo or in vitro assays known to the
skilled person.
6
In one embodiment, the GLP-1 receptor agonist is an insulinotropic analogue or
derivative of GLP-1(7-37) or an insulinotropic analogue or derivative of GLP-1(7-
36)NH2. In a particular embodiment, the GLP-1 receptor agonist is an
5 insulinotropic analogue or derivative of GLP-1(7-37). GLP-1(7-37) has the
sequence set out in SEQ ID NO. 1. GLP-1(7-36)NH2 has the sequence set out in
SEQ ID NO. 2.
An exemplary derivative of GLP-1(7-37) or GLP-1(7-36)NH2 bears one or more
10 lipophilic substituents attached to the parent peptide (GLP-1(7-37) or GLP-1(7-
36)NH2), optionally via a linker.
Another exemplary derivative is a fused derivative wherein the parent peptide
(GLP-1(7-37) or GLP-1(7-36)NH2) is fused to a protein, e.g. comprising serum
albumin or an antibody fragment e.g. the Fc portion of an antibody such as an
15 immunoglobulin G (IgG) e.g. IgG4 optionally via a linker.
Another exemplary derivative is one in which a simple amide (CONH2) is formed
of the C terminal COOH group.
20 Analogues of GLP-1(7-37) or GLP-1(7-36)NH2 comprise sequence changes and
exemplary analogues include exendin-4, exendin-3, an exendin-4 analogue, an
exendin-3 analogue, an exendin-4 derivative or an exendin-3 derivative. Exendin4 has the sequence set out in SEQ ID NO. 3. Exendin-3 has the sequence set
out in SEQ ID NO. 4.
25
In one embodiment, the GLP-1 receptor agonist comprises or more suitably
consists of a sequence sharing 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%,
90%, 95%, 98% or greater sequence identity with SEQ ID NO. 1 or SEQ ID NO.
2, suitably with SEQ ID. NO. 1. For example, there are up to 5 e.g. at most 1 or 2
30 amino acid changes as compared to SEQ ID NO. 1 or SEQ ID NO. 2.
For the purposes of comparing two related polypeptide sequences, the “%
sequence identity" between a first polypeptide sequence and a second
polypeptide sequence may be calculated using NCBI BLAST v2.0, using
35 standard settings for polypeptide sequences (BLASTP).
In one embodiment, the GLP-1 receptor agonist has or comprises the sequence
set out below (SEQ ID NO. 9):
H-X
1
-X
2
-G-T-F-T-S-D-X
3
-S-X
4
-X
5
-X
6
-E-X
7
-X
8
-A-X
9
-X
10
-X
11
-F-I-X
12
-W-L-X
13
-X
14
-GX
15 40
wherein
X
1
is A, G or S;
X
2
is E or D;
X
3
is V or L;
7
X
4
is S or K;
X
5
is Y or Q;
X
6
is L or M;
X
7
is G or E;
X
8
5 is Q or E;
X
9
is A or V;
X
10 is K or R;
X
11 is E or L;
X
12 is A or E;
X
13 10 is V or K;
X
14 is K, R or N; and
X
15 is R or G;
or is a derivative thereof such as a derivative in which a simple amide (CONH2)
is formed of the C terminal COOH group and/or a side chain bears a lipophilic
15 substituent, optionally via a linker.
In a further embodiment, the GLP-1 receptor agonist has or comprises the
sequence set out below:
[H-X
1
-X
2
-G-T-F-T-S-D-X
3
-S-X
4
-X
5
-X
6
-E-X
7
-X
8
-A-X
9
-X
10
-X
11
-F-I-X
12
-W-L-X
13
-X
14
-GX
15]n-X
16 20
wherein X
1
-X
15 are as described above for SED ID NO. 9;
n is 1 or 2; and
X
16 is absent, G, PSSGAPPPS (SEQ ID NO. 10), PSSGAPPSKKKKKK (SEQ ID
NO. 11), a serum albumin sequence or an FcIgG4 sequence;
25 or is a derivative thereof such as a derivative in which a simple amide (CONH2)
is formed of the C terminal COOH group and/or a side chain bears a lipophilic
substituent, optionally via a linker.
An exemplary serum albumin sequence is:
30 DAHKSEVAHRFKDLGEENFKALVLIAFAQYLQQCPFEDHVKLVNEVTEFAKTCV
ADESAENCDKSLHTLFGDKLCTVATLRETYGEMADCCAKQEPERNECFLQHKD
DNPNLPRLVRPEVDVMCTAFHDNEETFLKKYLYEIARRHPYFYAPELLFFAKRY
KAAFTECCQAADKAACLLPKLDELRDEGKASSAKQRLKCASLQKFGERAFKAW
AVARLSQRFPKAEFAEVSKLVTDLTKVHTECCHGDLLECADDRADLAKYICENQ
35 DSISSKLKECCEKPLLEKSHCIAEVENDEMPADLPSLAADFVESKDVCKNYAEA
KDVFLGMFLYEYARRHPDYSVVLLLRLAKTYETTLEKCCAAADPHECYAKVFDE
FKPLVEEPQNLIKQNCELFEQLGEYKFQNALLVRYTKKVPQVSTPTLVEVSRNL
GKVGSKCCKHPEAKRMPCAEDYLSVVLNQLCVLHEKTPVSDRVTKCCTESLVN
RRPCFSALEVDETYVPKEFNAETFTFHADICTLSEKERQIKKQTALVELVKHKPK
40 ATKEQLKAVMDDFAAFVEKCCKADDKETCFAEEGKKLVAASQAALGL (SEQ ID
NO. 12) which may optionally be conjugated to the rest of the GLP-1 receptor
agonist sequence via a linker sequence of e.g. up to 10 amino acids and may
optionally bear a C terminal extension of e.g. up to 10 amino acids.
8
An exemplary FcIgG4 sequence is:
GGGGGSGGGGSGGGGSAESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMI
SRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSV
LTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMT
5 KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVD
KSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (SEQ ID NO. 13) which may
optionally be conjugated to the rest of the GLP-1 receptor agonist sequence via a
linker sequence of e.g. up to 10 amino acids and may optionally bear a C
terminal extension of e.g. up to 10 amino acids.
10
Lipophilic substituents as mentioned above include acyl groups such as a straight
chain or branched fatty acid. A suitable acyl group typically has the formula
CH3(CH2)nCO-, wherein n = 4-38, for example 12-38, such as palmitoyl (i.e.
hexadecanoyl). The lipophilic substituent may be attached to the parent peptide
15 (i.e. main GLP-1 receptor agonist sequence) via a linker. A suitable linker is an
amino acid, wherein the lipophilic substituent may be attached to the amino acid
via the side chain of the amino acid, via the N-terminus of the amino acid, or via
the C-terminus of the amino acid. Any amino acid is potentially capable of acting
as a linker via connections to both its C-terminus and to its N-terminus. Amino
20 acids may also act as a linker via their side chain moiety. Such amino acids have
suitable side chain functionality for forming the linkage e.g. functionality which
can react with an acyl groups to form a stable bond. Suitable amino acids
include, but are not limited to, lysine, aspartic acid, glutamic acid, serine,
threonine, cysteine and tyrosine. In one embodiment the linker is a glutamic acid
25 residue and is suitably linked at one end via the carboxylic acid of its side chain
functionality (i.e. the ɣ/gamma-substituent) and at the other end by its N-terminus
or C-terminus, suitably via its N-terminus. The linker may also be a short peptide
sequence e.g. of 2-5 amino acids. In a particular embodiment, the lipophilic
substituent (with linker) is a γ-E-palmitoyl group (i.e. a γ-E-hexadecanoyl group).
30 In embodiments wherein the parent peptide is or comprises (GLP-1(7-37) or
GLP-1(7-36)NH2) (SEQ ID NO. 1 or SEQ ID NO. 2, respectively), suitably the
lipophilic substituent is attached to the parent peptide via the lysine residue at
position 26, optionally via a linker.
35 In embodiments wherein the parent peptide comprises the sequence set out in
SEQ ID NO. 9, the lipophilic substituent is suitably connected via residue X10
,
which is suitably a lysine residue, wherein the attachment is optionally via a
linker.
40 In one embodiment, the GLP-1 receptor agonist comprises one or more
asparagine or glutamine residues, particularly one or more glutamine residues,
and in particular said residue(s) is(are) within the sequence of SEQ ID NO. 9.
In one embodiment, the GLP-1 receptor agonist is selected from the group
consisting of albiglutide, dulaglutide, exenatide, liraglutide and lixisenatide.
9
Suitably, the GLP-1 receptor agonist is liraglutide or lixisenatide, in particular
liraglutide.
Albiglutide is a GLP-1 dimer fused to human albumin, having the sequence as set
5 out in SEQ ID NO. 5. Marketed under the tradenames Eperzan/Tanzeum, it has a
circulating half-life of 4-7 days.
Dulaglutide is a GLP-1 analogue consisting of GLP-1(7-37) covalently linked to
an Fc fragment of human IgG4, having the sequence as set out in SEQ ID NO. 6.
10 Marketed under the tradename Trulicity, it has a circulating half-life of
approximately 5 days.
Exenatide is a synthetic version of exendin-4, a GLP-1 analogue with 53%
sequence identity with GLP-1(7-36)NH2, having the sequence as set out in SEQ
15 ID NO. 3. Marketed under the tradenames Byetta/Bydureon, it has a circulating
half-life of 2.4 hours.
Liraglutide ((Arg(34), Lys(26)(N-(γ-Glu(N-palmitoyl)))-GLP-1(7-37)) is an acylated
GLP-1 analogue with 97% sequence identity with GLP-1(7-37), having the
20 sequence as set out in SEQ ID NO. 7. Marketed under the tradenames
Victoza/Saxenda, it has a circulating half-life of 13 hours.
Lixisenatide is an analogue of exendin-4, where proline is omitted at position 38
and the C-terminal has been modified to add six lysine residues, having the
25 sequence as set out in SEQ ID NO. 8. Marketed under the tradenames
Lyxumia/Adlyxin, it has a circulating half-life of 2-4 hours.
The concentration of GLP-1 receptor agonist in the composition is typically
between 10 µg/mL and 50 mg/mL, for example between 200 µg/mL and 10
30 mg/mL, or between 1 mg/mL and 10 mg/mL.
When the GLP-1 receptor agonist is albiglutide, suitably the concentration of
albiglutide in the composition is between 5 mg/mL and 200 mg/mL, for example
between 50 mg/mL and 150 mg/mL, such as around 100 mg/mL.
35
When the GLP-1 receptor agonist is dulaglutide, suitably the concentration of
dulaglutide in the composition is between 0.5 mg/mL and 20 mg/mL, for example
between 1 mg/mL and 5 mg/mL, such as around 1.5 mg/mL or around 3 mg/mL.
When the GLP-1 receptor agonist is exenatide, suitably the concentration of
40 exenatide in the composition is between 0.1 mg/mL and 0.5 mg/mL, for example
between 100 µg/mL and 500 µg/mL, such as around 250 µg/mL.
10
When the GLP-1 receptor agonist is liraglutide, suitably the concentration of
liraglutide in the composition is between 0.5 mg/mL and 20 mg/mL, for example
between 1 mg/mL and 10 mg/mL, such as around 6 mg/mL.
5 When the GLP-1 receptor agonist is lixisenatide, suitably the concentration of
lixisenatide in the composition is between 0.1 µg/mL and 250 µg/mL, for example
between 10 µg/mL and 50 µg/mL, such as around 17 µg/mL or around 33 µg/mL.
The composition of the invention contains multivalent anions as a stabilising
agent, wherein the multivalent anions have a charge of at least minus 2 (which
10 may also be written as “- 2” or “minus two”). The multivalent anions have a
charge of at least minus 2, such as minus 2 (“divalent anions”), minus 3 (“trivalent
anions”) or minus 4 (tetravalent anions”). In one embodiment, the multivalent
anions have a charge of minus 2 and are divalent anions. In another
embodiment, the multivalent anions have a charge of minus 3 and are trivalent
15 anions. In a further embodiment, the multivalent anions are a mixture of anions
having charge of minus 2 and anions having charge of minus 3, i.e. are a mixture
of divalent and trivalent anions. In another embodiment, the multivalent anions
are divalent anions, trivalent anions or a mixture thereof.
20 The multivalent anions are species which do not comprise any group capable of
forming a positive charge (e.g. by protonation) in the range of pH 4-9 at 25 °C.
Thus, the multivalent anions do not contain basic nitrogen centres, i.e. nitrogen
centres which are capable of being protonated. In particular, the multivalent
anions do not contain a quaternary ammonium group (i.e. a positively charged
25 tetra-substituted nitrogen atom). The multivalent anions do not contain
protonatable nitrogen centres with pKa between 5-10 or 3-11 at 25 °C. The
multivalent anions are not amino acids and particularly are not one of the 20
natural amino acids in L or D form or any mixture thereof (including a racemic
mixture). Thus, the multivalent anions are not glutamate, aspartate or a mixture
30 thereof. The multivalent anions are not peptides or proteins (i.e. molecules which
comprise two or more amino acid residues).
In one embodiment, the multivalent anions do not contain nitrogen atoms.
35 The multivalent anions are not nitrogen-containing chelating agents, and in
particular are not ethylenediaminetetraacetic acid (EDTA).
Suitably the multivalent anions have a molecular weight of less than 500 Da, for
example less than 400 Da, less than 300 Da or less than 200 Da.
40
If a molecular entity has more than one ionisable group, then it can exist in more
than one charged form (i.e. can exist as distinct species with differing charge). In
the context of the present invention each form with a different charge is
considered to be a separate species which may or may not be a suitable
11
multivalent anion within the context of the invention. For example, citric acid can
exist in the following charged states:
5 Only divalent citric-acid based species (“Species 2” in the figure above) with a
charge of minus 2 and trivalent citric-acid based species (“Species 3” in the figure
above) with a charge of minus 3 are multivalent anions according to the present
invention. Conversely, citric acid with a charge of 0, and monovalent citric acidbased species (“Species 1” in the figure above) with a charge of minus 1 are not
10 multivalent anions according to the present invention. In one embodiment, the
multivalent anions are divalent citrate anions. In another embodiment, the
multivalent anions are trivalent citrate anions. In a further embodiment, the
multivalent anions are selected from divalent citrate anions, trivalent citrate
anions and mixtures thereof. In a further embodiment, the multivalent anions are
15 a mixture of divalent and trivalent citrate anions.
Monovalent sulphate anions (also known as a bisulfate anions (HSO4
-
)) have a
charge of minus 1 therefore are not multivalent anions according to the present
invention. Divalent sulphate anions (SO4
2-
) have a charge of minus 2 and are
20 multivalent anions according to the present invention. In one embodiment the
multivalent anions are divalent sulphate anions.
Phosphoric acid ionizes to give the following species:
25 Only divalent phosphoric acid-based species (“Species 2” in the figure above)
with a charge of minus 2 and trivalent phosphoric acid-based species (“Species
3” in the figure above) with a charge of minus 3 are multivalent anions according
to the present invention. Conversely, phosphoric acid with a charge of 0, and
monovalent phosphoric acid-based species (“Species 1” in the figure above) with
30 a charge of minus 1 are not multivalent anions according to the present invention.
In one embodiment, the multivalent anions are divalent phosphate anions. In
another embodiment, the multivalent anions are trivalent phosphate anions. In a
further embodiment, the multivalent anions are selected from divalent phosphate
anions, trivalent phosphate anions and mixtures thereof. In a further embodiment,
35 the multivalent anions are a mixture of divalent and trivalent phosphate anions.
It should be noted that where multivalent anions with charge of at least minus 2
are present, this does not preclude other related species which are not
12
multivalent anions according to the invention from being present in the
composition. For example, in the case of the composition of the invention
containing divalent and/or trivalent phosphate anions (HPO4
2-
anions and/or PO4
3-
anions), phosphoric acid and/or monovalent phosphate anions (H2PO4
-
) may also
5 be present in the composition, but do not contribute to the “multivalent anions”
requirement of the invention.
Multivalent anions are conjugate bases of a conjugate acid-base pair. They can
be formed from the conjugate acid by addition of a strong base, resulting in a loss
10 of a proton. Therefore, in one embodiment the multivalent anions with charge of
at least minus 2 are added to the composition in the form of the conjugate acid
and pH is adjusted to a level which results in a loss of a proton and formation of
the conjugate base. For example, citric acid is added to the composition, followed
by adjustment of pH to a level where citrate multivalent anions are formed. More
15 commonly, the multivalent anions originate from a salt dissociation. Thus, in one
embodiment the multivalent anions with charge of at least minus 2 are added to
the composition in salt form, for example as a sodium salt, a potassium salt, a
calcium salt or a magnesium salt.
20 In one embodiment the multivalent anions are selected from divalent citrate
anions, trivalent citrate anions, divalent sulphate anions, divalent phosphate
anions, trivalent phosphate anions and mixtures thereof. In another embodiment,
the multivalent anions are selected from divalent citrate anions, trivalent citrate
anions, divalent sulphate anions, and mixtures thereof. In another embodiment,
25 the multivalent anions are selected from divalent citrate anions, trivalent citrate
anions, divalent phosphate anions, trivalent phosphate anions and mixtures
thereof. In another embodiment, the multivalent anions are selected from divalent
phosphate anions, trivalent phosphate anions, divalent sulphate anions and
mixtures thereof. Thus, reference to “multivalent anions” includes mixtures. In
30 one embodiment, the multivalent anions are a mixture of at least two different
multivalent anions having a charge of at least minus 2.
The multivalent anions are present in the composition at a total concentration of
at least 15 mM, for example at least 20 mM, at least 30 mM, at least 40 mM or at
35 least 50 mM. In one embodiment, the multivalent anions are present at a total
concentration of 15-150 mM, such as 15-120 mM, 20-120 mM, 20-100 mM, 20-
80 mM or around 50 mM. For the avoidance of doubt, where the multivalent
anions are a mixture of anions, the concentration is the total sum concentration of
every multivalent anion having a charge of at least minus 2 present in the
40 composition. Hence, the total concentration of multivalent anions may also be
described as the “combined concentration” of multivalent anions.
The concentration of multivalent anions in the composition is calculated based on
the concentration of the substance contributing the multivalent anions, the pH of
13
the solution and the pKa values of the ionisable groups on each species present
in the composition.
By way of illustration, phosphoric acid is a triprotic acid and dissociates in
5 solution in three steps as shown in the following formulae:
H3PO4 + H2O = H2PO4
-
+ H3O
+
Ka1 = [H2PO4
-
][H3O
+
] / [H3PO4]
H2PO4
-
+ H2O = HPO4
2-
+ H3O
+
Ka2 = [HPO4
2-
][H3O
+
] / [H2PO4
-
]
HPO4
2-
+ H2O = PO4
3-
+ H3O
+
10
Ka3 = [PO4
3-
][H3O
+
] / [HPO4
2-
]
The sum of the concentrations of the four phosphorus containing species after
equilibration at the appropriate pH is the same as the concentration of
15 phosphorus containing species added (this is a requirement for mass balance of
phosphorus). [H3O
+
] can be derived from the pH and the Ka1, Ka2 and Ka2 values
can be derived from the known values for pKa1, pKa2 and pKa3.
Thus, provided that the pH of the composition, the concentration of starting
20 materials and the pKa values of ionisable groups are known, a skilled person can
calculate the concentrations of each of the anions including the multivalent
anions.
For the avoidance of doubt, the multivalent anions cannot be the same species
25 as the GLP-1 receptor agonist.
In one embodiment, the aqueous solution composition of the invention does not
contain species which comprise groups capable of forming a positive charge in
the range of pH 4-9. In one embodiment, the aqueous solution composition of the
30 invention does not contain species comprising basic nitrogen centres i.e. nitrogen
centres which are capable of being protonated. For the avoidance of doubt, the
GLP-1 receptor agonist is not considered as a “species” in this context. In one
embodiment, the aqueous solution composition of the invention does not contain
species comprising protonatable nitrogen centres with pKa between 5-10 or 3-11
35 at 25 °C. Again for the avoidance of doubt, the GLP-1 receptor agonist is not
considered as a “species” in this context. In one embodiment, the aqueous
solution composition does not contain an amino acid. In another embodiment, the
aqueous solution composition does not contain a positively charged additive. In
one embodiment, the aqueous solution composition does not contain an additive
40 which bears a positive charge in the range of pH 4-9 at 25 °C. In one
embodiment, the aqueous solution composition does not contain propylene
glycol. In one embodiment, the aqueous solution composition does not contain
nitrogen-containing chelating agents, in particular EDTA.
14
The pH of the composition should be suitable for the particular GLP-1 receptor
agonist. For example, for a composition containing exenatide, the pH of the
composition is suitably between 4 and 5, for example about 4.5. For a
composition containing liraglutide, the pH of the composition is suitably between
5 7.5 and 8.5, for example about 8.1. In one embodiment, the pH of the
composition is between 4 and 9.
The compositions of the invention may additionally comprise a preservative such
as a phenolic or a benzylic preservative. The preservative is suitably selected
10 from the group consisting of phenol, m-cresol, chlorocresol, chlorophenol, benzyl
alcohol, propyl paraben, methyl paraben, benzalkonium chloride and
benzethonium chloride, in particular phenol, m-cresol and benzyl alcohol.
The concentration of preservative is typically 10-100 mM, for example 20-80 mM,
15 such as 25-50 mM. The optimal concentration of the preservative in the
composition is selected to ensure the composition passes the Pharmacopoeia
Antimicrobial Effectiveness Test (USP <51>, Vol. 32).
The composition may additionally comprise a surfactant. In one embodiment, the
20 surfactant is a non-ionic surfactant. In another embodiment, the surfactant is a
cationic surfactant.
A particularly suitable class of non-ionic surfactants is the polysorbates (fatty acid
esters of ethoxylated sorbitan), such as polysorbate 20. Polysorbate 20 is a
25 mono ester formed from oleic acid and polyoxyethylene (20) sorbitan in which the
number 20 indicates the number of oxyethylene groups in the molecule.
Polysorbate 20 is known under a range of brand names including in particular
Tween 20, and also Alkest TW 20. Other suitable polysorbates include
polysorbate 40, polysorbate 60 and polysorbate 80.
30
Another suitable class of non-ionic surfactants is the alkyl glycosides, especially
dodecyl maltoside. Other alkyl glycosides include dodecyl glucoside, octyl
glucoside, octyl maltoside, decyl glucoside, decyl maltoside, tridecyl glucoside,
tridecyl maltoside, tetradecyl glucoside, tetradecyl maltoside, hexadecyl
35 glucoside, hexadecyl maltoside, sucrose monooctanoate, sucrose mono
decanoate, sucrose monododecanoate, sucrose monotridecanoate, sucrose
monotetradecanoate and sucrose monohexadecanoate.
Another suitable class of non-ionic surfactants is block copolymers of
40 polyethylene glycol and polypropylene glycol, also known as poloxamers,
especially poloxamer 188, poloxamer 407, poloxamer 171 and poloxamer 185.
Poloxamers are also known under brand names Pluronics or Koliphors. For
example, poloxamer 188 is marketed as Pluronic F-68.
15
Another suitable class of non-ionic surfactants is alkyl ethers of polyethylene
glycol, especially those known under a brand name Brij, such as selected from
polyethylene glycol (2) hexadecyl ether (Brij 52), polyethylene glycol (2) oleyl
ether (Brij 93) and polyethylene glycol (2) dodecyl ether (Brij L4). Other suitable
5 Brij surfactants include polyethylene glycol (4) lauryl ether (Brij 30), polyethylene
glycol (10) lauryl ether (Brij 35), polyethylene glycol (20) hexadecyl ether (Brij 58)
and polyethylene glycol (10) stearyl ether (Brij 78).
Another suitable class of non-ionic surfactants are alkylphenyl ethers of
10 polyethylene glycol, especially 4-(1,1,3,3-tetramethylbutyl)phenyl-polyethylene
glycol, also known under a brand name Triton X-100.
Suitable cationic surfactants include benzalkonium and benzethonium salts. In
one embodiment, the cationic surfactant is selected from benzethonium salts e.g.
15 benzethonium halide such as benzethonium chloride. In another embodiment,
the cationic surfactant is selected from benzalkonium salts e.g. benzalkonium
halide such as benzalkonium chloride. In a further embodiment, the cationic
surfactant is a mixture of benzethonium salts and benzalkonium salts such as a
mixture of benzethonium chloride and benzalkonium chloride.
20
When included, the concentration of the surfactant in the formulation will typically
be in the range of 1-2000 µg/ml, e.g. 5-1000 µg/ml, e.g. 10-500 µg/ml, such as
10-200 µg/ml.
25 The compositions of the invention can have a wide range of osmolarity, and may
be hypotonic, isotonic or hypertonic. Suitably, the composition of the invention is
substantially isotonic. Suitably the osmolarity of the composition is selected to
minimize pain according to the route of administration e.g. upon injection.
Suitably the composition has an osmolarity of 50-1000 mOsm/L, such as 100-500
30 mOsm/L, for example about 300 mOsm/L.
The composition may additionally comprise a tonicity modifier, which may be
charged or uncharged. Examples of uncharged tonicity modifiers include glycerol,
mannitol, propylene glycol, trehalose, PEG300 and PEG400. When included, an
35 uncharged tonicity modifier is typically added at a concentration of 50-1000 mM,
for example 100-500 mM, such as about 300 mM. Examples of charged tonicity
modifiers include sodium chloride, sodium sulphate, sodium acetate, sodium
lactate, and amino acids such as glycine or arginine. When included, a charged
tonicity modifier is typically added at a concentration of 25-500 mM, for example
40 50-250 mM such as about 150 mM. In certain cases, if the concentration of
multivalent anions with a charge of at least minus 2 is sufficiently high then they
can also serve as a tonicity modifier.
16
The composition may comprise a buffer. Suitable buffers include acetate,
succinate, maleate, phosphate and TRIS. When included, the buffer is typically
present at a concentration of 0.5-50 mM, such as 1-20 mM, for example 2-5 mM.
In certain cases, the multivalent anions having a charge of at least minus 2 can
5 also serve as a buffer.
The composition may comprise additional active ingredients. In one embodiment,
the composition of the invention additionally comprises glucagon or a long acting
insulin. Examples of long acting insulins include insulin glargine and insulin
10 degludec. In one embodiment, the composition of the invention additionally
comprises peptide YY (peptide tyrosine tyrosine) or an analogue thereof. In one
embodiment, the GLP-1 receptor agonist is the only active ingredient in the
composition.
15 The following specific embodiments of the invention are envisaged. In one
embodiment is provided an aqueous solution composition comprising a GLP-1
receptor agonist which has or comprises the sequence set out in SEQ ID NO. 9
(wherein X1
-X
15 are as defined above) as an active ingredient, and multivalent
anions selected from divalent citrate anions, trivalent citrate anions, divalent
20 sulphate anions, divalent phosphate anions, trivalent phosphate anions and
mixtures thereof, as stabilising agent, wherein the total concentration of
multivalent anions in the composition is at least 15 mM, at least 20 mM, at least
30 mM, at least 40 mM or at least 50 mM. In another embodiment is provided an
aqueous solution composition comprising a GLP-1 receptor agonist selected from
25 albiglutide, dulaglutide, exenatide, liraglutide and lixisenatide as an active
ingredient, and multivalent anions selected from divalent citrate anions, trivalent
citrate anions, divalent sulphate anions, divalent phosphate anions, trivalent
phosphate anions and mixtures thereof, as stabilising agent, wherein the total
concentration of multivalent anions in the composition is at least 15 mM, at least
30 20 mM, at least 30 mM, at least 40 mM or at least 50 mM. In another
embodiment is provided an aqueous solution composition comprising a GLP-1
receptor agonist which is liraglutide as an active ingredient, and multivalent
anions selected from divalent citrate anions, trivalent citrate anions and mixtures
thereof as stabilising agent wherein the total concentration of multivalent anions
35 in the composition is at least 15 mM, at least 20 mM, at least 30 mM, at least 40
mM or at least 50 mM. In another embodiment is provided an aqueous solution
composition comprising a GLP-1 receptor agonist which is liraglutide as an active
ingredient, and multivalent anions which are divalent sulphate anions as
stabilising agent wherein the total concentration of multivalent anions in the
40 composition is at least 15 mM, at least 20 mM, at least 30 mM, at least 40 mM or
at least 50 mM. In another embodiment is provided an aqueous solution
composition comprising a GLP-1 receptor agonist which is liraglutide as an active
ingredient, and multivalent anions which are divalent sulphate anions as
stabilising agent, at a concentration of at least 15 mM, at least 20 mM, at least 30
17
mM, at least 40 mM or at least 50 mM. In another embodiment is provided an
aqueous solution composition comprising a GLP-1 receptor agonist which is
liraglutide as an active ingredient, and multivalent anions selected from divalent
phosphate anions, trivalent phosphate anions and mixtures thereof as stabilising
5 agent wherein the total concentration of multivalent anions in the composition is
at least 15 mM, at least 20 mM, at least 30 mM, at least 40 mM or at least 50
mM..
The presently claimed invention derives from the surprising observation that
10 compositions containing a GLP-1 receptor agonist as active ingredient, in
particular liraglutide, are stabilized by the addition of multivalent anions,
particularly divalent and trivalent anions. Having recognized the need to improve
the stability of currently marketed formulations containing GLP-1 receptor
agonists, the present inventors began their investigations by using a composition
15 based on Victoza (liraglutide), wherein the tonicity modifier propylene glycol was
replaced with various alternative tonicity modifiers. Stability (primarily chemical
stability) was determined with respect to product purity assessed by reversedphase HPLC. As explained in Example 1, replacement of the propylene glycol
with mannitol, trehalose or sodium chloride was observed to have minimal impact
20 on stability of the liraglutide. The addition of histidine was observed to impair the
stability of liraglutide. The tonicity modifier propylene glycol was then replaced
with various multivalent anions. As can be seen from Example 2 and Figures 3-5,
the presence of multivalent anions with a charge of at least minus 2 led to a
considerable improvement in the stability of liraglutide following storage at 30 °C
25 for 9 weeks and 14 weeks.
GLP-1 receptor agonists share very similar instability pathways. Whilst the
physical instability of these products comprises formation of soluble aggregates
and insoluble aggregates (non-specifically structured particles and structured
30 amyloid fibrils), the sites of chemical instability are limited to asparagine (N) or
glutamine (Q) residues that are prone to the formation of cyclic imide structures,
which are in turn cleaved to asparte or isoaspartate residues (in the case of
asparagine) or glutamate or isoglutamate residues (in the case of glutamine).
These processes are referred to as aspartate/glutamate deamidation. Although
35 the cyclic imide structure is an intermediate in this process, it is often a relatively
stable impurity for this type of therapeutic peptide. The chemically related
impurities of GLP-1 receptor agonists are thus typically represented by cyclic
imides, and molecules where glutamine/asparagine have turned into the
respective acids or iso-acids. Without being bound by theory, it is believed that
40 the surprising stabilising effect of multivalent anions is due at least in part to their
binding to the structure of the GLP-1 receptor agonist, resulting in formation of a
tighter peptide structure which in turn limits the exposure of the deamidation
sights to the solvent, thus limiting the rate of deamidation. Given the high degree
of homology shared amongst the GLP-1 receptor agonists, very similar
18
interactions are expected between their respective structures and the multivalent
anions. Consequently, it is considered likely that the improvement in stability
observed for liraglutide upon addition of multivalent anions with charge of at least
minus 2 will also be observed for GLP-1 receptor agonists generally, in particular
5 exenatide, dulaglutide, lixisenatide and albiglutide.
Chemical stability of a GLP-1 receptor composition can be evaluated by reversed
phase HPLC (RP-HPLC) to determine the proportion of chemically related GLP-1
receptor agonist species (i.e. species generated during storage or other stress
10 conditions by chemical modification of GLP-1 receptor agonist, including
deamidation or cyclic imide formation) as described in the General Procedures.
This type of RP-HPLC experiment can be used to determine the purity of the
GLP-1 receptor agonist species i.e. the amount of unaltered GLP-1 receptor
agonist (e.g. as a % by total weight of the original GLP-1 receptor agonist)
15 following storage or other stress conditions.
Suitably, a composition of the invention retains at least 90%, e.g. at least 91%, at
least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at
least 98% or at least 99% purity (i.e. native GLP-1 receptor agonist (by total
20 weight of GLP-1 receptor agonist in the composition at time T = 0)) following
storage at 15-30 °C e.g. 30 °C, for at least 3 weeks, for example for 3 weeks, 4
weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12
weeks, 13 weeks or 14 weeks.
25 Suitably, a composition of the invention retains at least 95%, e.g. at least 96%, at
least 97%, at least 98% or at least 99% purity (i.e. native GLP-1 receptor agonist
(by total weight of GLP-1 receptor agonist in the composition at time T = 0))
following storage for 9 weeks at 30 °C.
30 Suitably, a composition of the invention retains at least 90%, e.g. at least 91%, at
least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at
least 98% or at least 99% purity (i.e. native GLP-1 receptor agonist (by total
weight of GLP-1 receptor agonist in the composition at time T = 0)) following
storage for 14 weeks at 30 °C.
35
Suitably, a composition of the invention retains at least 90%, e.g. at least 91%, at
least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at
least 98% or at least 99% purity (i.e. native GLP-1 receptor agonist (by total
weight of GLP-1 receptor agonist in the composition at time T = 0)) following
40 storage at 32 °C or higher, for example at 32 °C, 34 °C, 35 °C, 36 °C, 37 °C, 38
°C or 40 °C, for at least 3 weeks, for example for 3 weeks, 4 weeks, 5 weeks, 6
weeks, 7 weeks, 8 weeks, 10 weeks, 12 weeks or 14 weeks.
19
Suitably, a composition of the invention retains at least 90%, e.g. at least 91%, at
least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at
least 98% or at least 99% purity (i.e. native GLP-1 receptor agonist (by total
weight of GLP-1 receptor agonist in the composition at time T = 0)) following
5 storage at 25 °C for at least 3 months, such as 3 months, 6 months, 12 months or
24 months; and also retains at least 90%, e.g. at least 91%, at least 92%, at least
93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at
least 99% purity (i.e. native GLP-1 receptor agonist (by total weight of GLP-1
receptor agonist in the composition at time T = 0)) following an in-use period of
10 28 days at 30 °C, starting immediately after the end of the storage period.
Suitably, a composition of the present invention should exhibit an increase in
chemically related GLP-1 receptor agonist species during storage which is at
least 10% lower, preferably at least 25% lower, more preferably at least 50%
15 lower, than a composition lacking multivalent anions having charge of at least
minus 2 as a stabilising agent, but otherwise identical, following storage under
the same conditions and length of time.
Suitably, the proportion of total chemically related GLP-1 receptor agonist
20 species remains below 10% (by weight), e.g. below 9%, 8%, 7%, 6%, 5%, 4%,
3%, 2%, or below 1% during storage at 15-30 °C e.g. 30 °C, for at least 4 weeks,
for example 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks,
11 weeks, 12 weeks, 13 weeks or 14 weeks.
25 Suitably, the proportion of total chemically related GLP-1 receptor agonist
species remains below 5% (by weight), e.g. below 4%, 3%, 2%, or below 1%
during storage at 30 °C for 9 weeks.
Suitably, the proportion of total chemically related GLP-1 receptor agonist
30 species remains below 10% (by weight), e.g. below 9%, 8%, 7%, 6%, 5%, 4%,
3%, 2%, or below 1% during storage at 30 °C for 14 weeks.
Suitably, the proportion of total chemically related GLP-1 receptor agonist
species remains below 5% (by weight), e.g. below 4%, 3%, 2%, or 1%, during
35 storage for at least 3 weeks such as at least 4 weeks, at temperatures greater
than 30 °C, for example during storage for 3 weeks, 4 weeks, 5 weeks, 6 weeks,
7 weeks, 8 weeks or 9 weeks at 32 °C, 34 °C, 35 °C, 37 °C, 38 °C or 40 °C.
Suitably, the proportion of total chemically related GLP-1 receptor agonist
40 species remains below 10% (by weight), e.g. below 9%, 8%, 7%, 6%, 5%, 4%,
3%, 2%, or 1% during storage at least 32 °C or higher, for example at 32 °C,
34 °C, 35 °C, 37 °C, 38 °C or 40 °C, for at least 4 weeks, for example for 4
weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12
weeks, 13 weeks or 14 weeks.
20
In another aspect of the invention, there is provided the use of multivalent anions
having a charge of at least minus 2 and suitably wherein the total concentration
of multivalent anions in the composition having a charge of at least minus 2 is at
least 15 mM, for stabilizing an aqueous solution composition of a GLP-1 receptor
5 agonist. In particular, the chemical stability of the GLP-1 receptor agonist is
improved.
In a further aspect of the invention, there is provided a method of improving the
stability (i.e. chemical and/or physical stability) of an aqueous solution
10 composition comprising a GLP-1 receptor agonist as an active ingredient which
comprises adding multivalent anions having a charge of at least minus 2 to the
composition and suitably wherein the total concentration of multivalent anions in
the composition having a charge of at least minus 2 is at least 15 mM. In
particular, the chemical stability of the GLP-1 receptor agonist is improved.
15
In a further aspect of the invention, there is provided a method for increasing the
shelf-life of an aqueous solution composition comprising a GLP-1 receptor
agonist as an active ingredient which comprises adding multivalent anions having
a charge of at least minus 2 to the composition and suitably wherein the total
20 concentration of multivalent anions in the composition having a charge of at least
minus 2 is at least 15 mM.
The physical stability of a GLP-1 receptor agonist composition refers to the
tendency of the agonist molecule to form soluble aggregates or insoluble
25 aggregates (particles or fibrils) due to, inter alia, destabilizing interactions with
surfaces and interfaces, and temperature fluctuations. Physical stability of the
GLP-1 receptor agonist composition can be evaluated by methods known in the
art, including by visual inspection, size exclusion chromatography (SEC),
electrophoresis, analytical ultracentrifugation, light scattering, dynamic light
30 scattering, static light scattering or field flow fractionation. Fibril formation can be
assessed by Thioflavin T (ThT) assay. In the present Examples, all aqueous
solution compositions of the invention were observed to have good physical
stability as no aggregates (soluble or insoluble) were observed.
35 The chemical stability of a GLP-1 receptor composition refers in particular to the
stability with respect to formation of chemically related GLP-1 receptor agonist
species (i.e. species generated during storage or other stress conditions by
chemical modification of GLP-1 receptor agonist, including deamidation or cyclic
imide formation).
40
Suitably the composition of the invention remains as a clear solution for a longer
period of time compared to the current commercially available GLP-1 receptor
agonist products, allowing for a longer in-use period. Thus, in one embodiment,
the composition of the invention remains as a clear solution during storage at 15-
21
30 °C e.g. at 30 °C for longer than 4 weeks, for example for 5 weeks, 6 weeks, 7
weeks, 8 weeks, 9 weeks, 10 weeks or 12 weeks. By “clear solution” it is meant
that no visible precipitation is observed during storage.
5 Suitably the composition of the invention remains as a clear solution when
exposed to temperatures which are higher than those recommended for the
current commercially available GLP-1 receptor agonist products. Thus, in one
embodiment, the composition of the invention remains as a clear solution during
storage for 4 weeks, at temperatures greater than 30 °C, for example during
10 storage for 4 weeks at 32 °C, at 34 °C, at 35 °C, at 37 °C, at 38 °C or at 40 °C.
In one embodiment, the composition according to the invention is a clear solution
with low viscosity (e.g. dynamic viscosity of less than 20 cP, such as less than 10
cP, e.g. less than 5 cP at 25 °C measured using a microfluidics capillary
extrusion viscometer, such as m-VROC™, RheoSense Inc.).
15
Suitably the composition of the invention has improved storage stability at
increased temperature, while maintaining the in-use stability. Thus, in one
embodiment, the composition of the invention remains as a clear solution during
storage at 25 °C for at least 3 months, for example 3 months, 6 months, 12
20 months or 24 months; and also remains as a clear solution during an in-use
period of 14 days or 28 days at 30 °C, starting immediately after the end of the
storage period.
In one embodiment, a composition of the present invention exhibits an increase
25 in high molecular weight species during storage which is at least 10% lower,
preferably at least 25% lower, more preferably at least 50% lower, than a
composition lacking multivalent anions having a charge of at least minus 2 as a
stabilising agent, but otherwise identical (and particularly lacking multivalent
anions in the composition having a charge of at least minus 2 at a concentration
30 of at least 15 mM) following storage under the same conditions and length of
time.
The composition of the invention is a therapeutic composition (i.e. a composition
for use in therapy), and is particularly suitable for the treatment or prevention of a
35 disease or disorder caused by, associated with and/or accompanied by
disturbances in carbohydrate and/or lipid metabolism, e.g. for the treatment or
prevention of type 1 diabetes, type 2 diabetes, hyperglycaemia, impaired glucose
tolerance, obesity or metabolic syndrome. Thus, in one embodiment is provided a
method of treating or preventing a disease or disorder caused by, associated with
40 and/or accompanied by disturbances in carbohydrate and/or lipid metabolism,
e.g. for the treatment or prevention of type 1 diabetes, type 2 diabetes,
hyperglycaemia, impaired glucose tolerance, obesity or metabolic syndrome,
which comprises administering to a subject in need thereof a therapeutically
effective amount of a composition as described herein. There is also provided a
22
composition as described herein for use as a pharmaceutical, especially for use
in treating or preventing diseases or disorders caused by, associated with and/or
accompanied by disturbances in carbohydrate and/or lipid metabolism, e.g. for
the treatment or prevention of type 1 diabetes, type 2 diabetes, hyperglycaemia,
5 impaired glucose tolerance, obesity or metabolic syndrome.
The therapeutic compositions of the invention are suitably intended for a daily
administration, particularly for treatment of type 2 diabetes. The compositions
may also be used for less frequent administration, such as twice-weekly, weekly
10 or monthly administration.
There is also provided a container, for example made of plastics or glass,
containing one dose or a plurality of doses of the composition as described
herein. The container can be for example, a vial, or a cartridge designed to be a
15 replaceable item for use with an injection device.
The compositions of the invention may suitably be packaged for injection,
especially sub-cutaneous or intramuscular injection. Sub-cutaneous injection is
preferred. Injection may be by conventional syringe or more preferably via a pen
20 device adapted for use by diabetic subjects. Exemplary pen devices include
OptiClick®
, SoloSTAR®
and KwikPen®
.
An aspect of the invention is an injection device, particularly a device adapted for
subcutaneous or intramuscular injection, for single or multiple-use comprising a
25 container containing one dose or a plurality of doses of the composition of the
invention together with an injection needle. In an embodiment the container is a
replaceable cartridge which contains a plurality of doses. In an embodiment, the
needle is replaceable e.g. after each occasion of use. In one embodiment, the
injection device is in the form of a pen. In one embodiment, the injection device is
30 in the form of a pump. In one embodiment, the injection device comprises a
pump.
Another aspect of the invention is a medical device comprising a reservoir
comprising a plurality of doses of the composition of the invention and a pump
35 adapted for automatic or remote operation such that upon automatic or remote
operation one or more doses of the composition of the invention is administered
to the body e.g. subcutaneously or intramuscularly. Such devices may be worn
on the outside of the body or implanted in the body.
40 Further aspects of the invention include:
An aqueous solution composition comprising liraglutide as an active ingredient
and ions selected from citrate ions, phosphate ions and sulphate ions (e.g.
contributed by added sodium citrate, sodium phosphate and/or sodium sulphate)
at a concentration of at least 15 mM, 20 mM, 30 mM, 40 mM, 50 mM, or 100 mM
23
wherein the pH of the composition is 8-8.5 e.g. 8.1-8.2. For example, the
concentration of ions selected from citrate ions, phosphate ions and sulphate
ions does not exceed 150 mM e.g. does not exceed 120 mM.
5 An aqueous solution composition comprising liraglutide as an active ingredient
and citrate ions (e.g. contributed by added sodium citrate) at a concentration of at
least 15 mM, 20 mM, 30 mM, 40 mM, 50 mM, or 100 mM wherein the pH of the
composition is 8-8.5 e.g. 8.1-8.2. For example, the concentration of citrate ions
does not exceed 150 mM e.g. does not exceed 120 mM. The composition may
10 contain phosphate ions (e.g. contributed by added sodium phosphate) at a
concentration of e.g. 5-10 mM.
An aqueous solution composition comprising liraglutide as an active ingredient
and sulphate ions (e.g. contributed by added sodium sulphate) at a concentration
15 of at least 15 mM, 20 mM, 30 mM, 40 mM, 50 mM, or 100 mM wherein the pH of
the composition is 8-8.5 e.g. 8.1-8.2. For example, the concentration of sulphate
ions does not exceed 150 mM e.g. does not exceed 120 mM. The composition
may contain phosphate ions (e.g. contributed by added sodium phosphate) at a
concentration of e.g. 5-10 mM.
20
An aqueous solution composition comprising liraglutide as an active ingredient
and phosphate ions (e.g. contributed by added sodium phosphate) at a
concentration of at least 15 mM, 20 mM, 30 mM, 40 mM, 50 mM, or 100 mM
wherein the pH of the composition is 8-8.5 e.g. 8.1-8.2. For example, the
25 concentration of phosphate ions does not exceed 150 mM e.g. does not exceed
120 mM.
GLP-1 receptor agonist-containing compositions according to the invention are
expected to have one or more of the advantages of:
30  good stability, in particular chemical stability, during the in-use period at
temperatures up to 40 °C, e.g. up to 30 °C;
 good stability, in particular chemical stability, during an extended in-use
period e.g. up to 12 weeks;
 good storage stability, in particular chemical storage stability, at an
35 increased temperature e.g. 20-25 °C whilst retaining good in-use stability,
in particular chemical in-use stability;
Compositions according to the invention are expected to have good chemical
and/or physical stability as described herein, in particular good chemical stability.
40
EXAMPLES
General procedures
Reversed phase high-performance liquid chromatography (RP-HPLC)
24
Ultra-high performance reverse phase chromatography was performed using the
Waters ACQUITY H-class Bio UPLC®
system with a 1.7 μm Ethylene Bridged
Hybrid particle, 130 Å pore resin trifunctionally immobilised with a C18 ligand in a
50 mm by 2.1mm column. Insulin samples were bound in an 82% w/v Na2SO4,
5 18% v/v acetonitrile, pH 2.3 mobile phase and eluted in 50% w/v Na2SO4, 50%
v/v acetonitrile gradient flow. 2 μl of sample was acidified with 0.01M HCl and
analysed at 0.61 mL/min, with 214 nm UV detection. All analyses were performed
at 40 °C.
10 Example 1 (comparative) – Effect of additives on the stability of liraglutide
The effect of various tonicity modifiers on the stability of liraglutide was
investigated by replacing propylene glycol in the composition of the marketed
liraglutide product (Victoza) with two alternative uncharged tonicity modifiers
(mannitol and trehalose), one charged tonicity modifier (sodium chloride) and one
15 amino acid (histidine) and assessing purity by RP-HPLC (see General
Procedures) following storage at 30 °C. The remainder of the composition was
identical to that of the Victoza product (liraglutide (6 mg/ml), sodium phosphate (8
mM), phenol (58.4 mM), pH 8.15). It was shown (Table 1 and Figure 1) that
replacing propylene glycol with mannitol, trehalose or sodium chloride had
20 minimal impact on the stability of liraglutide. Histidine, both at 10 mM and at 50
mM concentration, impaired the stability of liraglutide (Table 1 and Figure 2).
Table 1: Purity of liraglutide assessed by visual assessment following storage at
30 °C for 9 and 14 weeks. All formulations were adjusted to pH 8.15 and
25 contained liraglutide (6 mg/ml), sodium phosphate (8 mM), phenol (58.4 mM) and
additive(s) specified in the Table. Purity was assessed by RP-HPLC.
Additive MVA*
conc.
(mM)
Purity (assessed as % RPHPLC main peak)
0 weeks 9
weeks
14
weeks
Propylene glycol (184 mM) 7.3 99.75 94.05 90.91
Mannitol (184 mM) 7.3 99.70 94.15 90.02
Trehalose (184 mM) 7.3 99.79 94.01 90.39
Sodium chloride (150 mM) 7.3 99.69 94.45 89.33
Histidine (10 mM) + Mannitol (184
mM)
7.3 99.69 92.39 80.98
Histidine (50 mM) + Mannitol (184
mM)
7.3 99.81 90.62 78.53
*Multivalent anions having a charge of at least minus 2 (based on pKa values of
phosphate of 2.2, 7.2 and 12.3 (25 °C); CRC Handbook of Chemistry and
Physics, 79th Edition, 1998, D. R. Lide)
30
25
Example 2 – Effect of multivalent anions on the stability of liraglutide
The effect of multivalent anions on the stability of liraglutide was investigated by
adding the various anions to a composition comprising liraglutide (6 mg/ml),
sodium phosphate (8 mM), phenol (58.4 mM) at pH 8.15, and assessing purity by
5 RP-HPLC following storage at 30 °C. The effect was investigated in the presence
of mannitol as a tonicity modifier. In some cases, particularly if the concentration
of the multivalent anion was high, the effect was also investigated in the absence
of mannitol. Stability of the compositions containing the multivalent anions was
compared with that of the compositions containing mannitol only or propylene
10 glycol only (i.e. the composition of the marketed Victoza product).
It was shown (Table 2 and Figure 3) that the presence of citrate anion (from
sodium citrate) led to a considerable improvement in stability of liraglutide. The
effect appeared to be strongest at 50 mM citrate, but was also very apparent at
15 citrate concentrations of 10 mM and 100 mM. At 100 mM citrate concentration,
the effect appeared to be independent of the presence of mannitol. Similarly, the
presence of higher concentration of phosphate anion (from sodium phosphate)
had a stabilizing effect on liraglutide (Figure 4). Whilst the control formulations
(i.e. the first two formulations in Table 2) also contained a small amount of
20 phosphate anion as a buffer, the stability was improved by increasing the
concentration to either 50 mM or 100 mM. The presence of sulphate anion (from
sodium sulphate) also improved the stability of liraglutide, particularly at 50 mM
concentration (Figure 5). The stabilizing effect of 100 mM sulphate anion was
markedly less strong than that of 50 mM sulphate anion, but it was still
25 measurable versus the multivalent anion free compositions.
Table 2: Purity of liraglutide assessed by visual assessment following storage at
30 °C for 9 and 14 weeks. All formulations were adjusted to pH 8.15 and
contained liraglutide (6 mg/ml), sodium phosphate (8 mM), phenol (58.4 mM) and
30 additive(s) specified in the Table. Purity was assessed by RP-HPLC.
Additive MVA**
conc.
(mM)
Purity (assessed as % RPHPLC main peak)
0
weeks
9
weeks
14
weeks
Propylene glycol (184 mM) 7.3 99.75 94.05 90.91
Mannitol (184 mM) 7.3 99.70 94.15 90.02
Sodium citrate (10 mM) + Mannitol
(184 mM)
17.3 99.68 96.30 94.77
Sodium citrate (50 mM) + Mannitol
(184 mM)
57.3 99.75 96.83 95.66
Sodium citrate (100 mM) + Mannitol
(184 mM)
107.3 99.75 96.37 94.86
Sodium citrate (100 mM) 107.3 99.74 95.89 94.26
26
Phosphate (42 mM)* + Mannitol (184
mM)
45.6 99.83 96.01 93.34
Phosphate (92 mM)* + Mannitol (184
mM)
91.2 99.72 96.02 93.52
Phosphate (92 mM)* 91.2 99.76 96.12 93.43
Sodium sulphate (50 mM) + Mannitol
(184 mM)
57.3 99.71 96.33 94.61
Sodium sulphate (100 mM) + Mannitol
(184 mM)
107.3 99.70 95.42 92.77
Sodium sulphate (100 mM) 107.3 99.68 95.12 92.11
* This concentration does not include 8 mM phosphate that was added to all
formulations.
**Multivalent anion having a charge of at least minus 2 (based on pKa values of
phosphoric acid of 2.2, 7.2 and 12.3 (25 °C), pKa values of citric acid of 3.1, 4.8
5 and 6.4 (25 °C) and pKa values of sulphuric acid of -3.0 and 2.0 (25 °C); pKa
values obtained from CRC Handbook of Chemistry and Physics, 79th Edition,
1998, D. R. Lide (phosphoric acid and sulphuric acid) and Merck Index (citric
acid)).
10 Unless otherwise stated, pH, pKa and other physical parameters are determined
at 25 °C.
Throughout the specification and the claims which follow, unless the context
requires otherwise, the word ‘comprise’, and variations such as ‘comprises’ and
15 ‘comprising’, will be understood to imply the inclusion of a stated integer, step,
group of integers or group of steps but not to the exclusion of any other integer,
step, group of integers or group of steps.
All patents, patent applications and references mentioned throughout the
20 specification of the present invention are herein incorporated in their entirety by
reference.
The invention embraces all combinations of preferred and more preferred groups
and suitable and more suitable groups and embodiments of groups recited
25 above.
27
SEQUENCE LISTING
SEQ ID NO. 1 (GLP-1(7-37)): HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG
SEQ ID NO. 2 (GLP-1(7-36)NH2): HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRNH2
5 SEQ ID NO. 3 (exendin-4/Exexatide):
HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS-NH2
SEQ ID NO. 4 (exendin-3):
HSDGTFTSDLSKQMEEEAVRLFIEWLKNGGPSS
GAPPPS-NH2
10 SEQ ID NO. 5 (Albiglutide):
HGEGTFTSDVSSYLEGQAAKEFIAWLVKGRHGEGTFTSDVSSYLEGQAAKEFIA
WLVKGRDAHKSEVAHRFKDLGEENFKALVLIAFAQYLQQCPFEDHVKLVNEVT
EFAKTCVADESAENCDKSLHTLFGDKLCTVATLRETYGEMADCCAKQEPERNE
CFLQHKDDNPNLPRLVRPEVDVMCTAFHDNEETFLKKYLYEIARRHPYFYAPEL
15 LFFAKRYKAAFTECCQAADKAACLLPKLDELRDEGKASSAKQRLKCASLQKFGE
RAFKAWAVARLSQRFPKAEFAEVSKLVTDLTKVHTECCHGDLLECADDRADLA
KYICENQDSISSKLKECCEKPLLEKSHCIAEVENDEMPADLPSLAADFVESKDVC
KNYAEAKDVFLGMFLYEYARRHPDYSVVLLLRLAKTYETTLEKCCAAADPHECY
AKVFDEFKPLVEEPQNLIKQNCELFEQLGEYKFQNALLVRYTKKVPQVSTPTLV
20 EVSRNLGKVGSKCCKHPEAKRMPCAEDYLSVVLNQLCVLHEKTPVSDRVTKCC
TESLVNRRPCFSALEVDETYVPKEFNAETFTFHADICTLSEKERQIKKQTALVEL
VKHKPKATKEQLKAVMDDFAAFVEKCCKADDKETCFAEEGKKLVAASQAALGL
SEQ ID NO. 6 (Dulaglutide):
HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSAESKY
25 GPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQF
NWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKG
LPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALH
NHYTQKSLSLSLG
30 SEQ ID NO. 7 (Liraglutide): HAEGTFTSDVSSYLEGQAAK(ɣ-Epalmitoyl)EFIAWLVRGRG
SEQ ID NO. 8 (Lixisenatide):
HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPSKKKKKK-NH2
SEQ ID NO. 9 (artificial sequence): H-X
1
-X
2
-G-T-F-T-S-D-X
3
-S-X
4
-X
5
-X
6
-E-X
7
-X
8
-
A-X
9
-X
10
-X
11
-F-I-X
12
-W-L-X
13
-X
14
-G-X
15 35
SEQ ID NO. 10 (artificial sequence): PSSGAPPPS
SEQ ID NO. 11 (artificial sequence): PSSGAPPSKKKKKK
SEQ ID NO. 12 (exemplary serum albumin sequence):
DAHKSEVAHRFKDLGEENFKALVLIAFAQYLQQCPFEDHVKLVNEVTEFAKTCV
40 ADESAENCDKSLHTLFGDKLCTVATLRETYGEMADCCAKQEPERNECFLQHKD
DNPNLPRLVRPEVDVMCTAFHDNEETFLKKYLYEIARRHPYFYAPELLFFAKRY
KAAFTECCQAADKAACLLPKLDELRDEGKASSAKQRLKCASLQKFGERAFKAW
AVARLSQRFPKAEFAEVSKLVTDLTKVHTECCHGDLLECADDRADLAKYICENQ
DSISSKLKECCEKPLLEKSHCIAEVENDEMPADLPSLAADFVESKDVCKNYAEA
28
KDVFLGMFLYEYARRHPDYSVVLLLRLAKTYETTLEKCCAAADPHECYAKVFDE
FKPLVEEPQNLIKQNCELFEQLGEYKFQNALLVRYTKKVPQVSTPTLVEVSRNL
GKVGSKCCKHPEAKRMPCAEDYLSVVLNQLCVLHEKTPVSDRVTKCCTESLVN
RRPCFSALEVDETYVPKEFNAETFTFHADICTLSEKERQIKKQTALVELVKHKPK
5 ATKEQLKAVMDDFAAFVEKCCKADDKETCFAEEGKKLVAASQAALGL
SEQ ID NO. 13 (exemplary FcIgG4 sequence):
GGGGGSGGGGSGGGGSAESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMI
SRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSV
LTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMT
10 KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVD
KSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG
29
We claim:
1. An aqueous solution composition comprising a GLP-1 receptor agonist as
an active ingredient and multivalent anions having a charge of at least minus 2 as
stabilising agent, wherein the total concentration of multivalent anions in the
5 composition having a charge of at least minus 2 is at least 15 mM.
2. An aqueous solution composition according to claim 1, wherein the GLP-1
receptor agonist is an insulinotropic analogue or derivative of GLP-1(7-37) (SEQ
ID NO. 1) or an insulinotropic analogue or derivative of GLP-1(7-36)NH2 (SEQ ID
10 NO. 2).
3. An aqueous solution composition according to claim 1 or claim 2, wherein
the GLP-1 receptor agonist has or comprises the sequence set out below (SEQ
ID NO. 9):
H-X
1
-X
2
-G-T-F-T-S-D-X
3
-S-X
4
-X
5
-X
6
-E-X
7
-X
8
-A-X
9
-X
10
-X
11
-F-I-X
12
-W-L-X
13
-X
14 15 -GX
15
wherein
X
1
is A, G or S; X2
is E or D; X3
is V or L; X4
is S or K; X5
is Y or Q; X6
is L or M;
X
7
is G or E; X8
is Q or E; X9
is A or V; X10 is K or R; X11 is E or L; X12 is A or E;
X
13 is V or K; X14 is K, R or N; and X15 20 is R or G;
or is a derivative thereof such as a derivative in which a simple amide (CONH2)
is formed of the C terminal COOH group and/or a side chain bears a lipophilic
substituent, optionally via a linker.
25 4. An aqueous solution composition according to any one of claims 1 to 3,
wherein the GLP-1 receptor agonist is selected from the group consisting of
albiglutide, dulaglutide, exenatide, liraglutide and lixisenatide.
5. An aqueous solution composition according to claim 4, wherein the GLP-1
30 receptor agonist is liraglutide or lixisenatide
6. An aqueous solution composition according to claim 5, wherein the GLP-1
receptor agonist is liraglutide.
35 7. An aqueous solution composition according to any one of claims 1 to 6,
wherein the concentration of GLP-1 receptor agonist in the composition is
between 10 µg/mL and 50 mg/mL, such as between 200 µg/mL and 10 mg/mL,
or between 1 mg/mL and 10 mg/mL.
40 8. An aqueous solution composition according to any one of claims 1 to 7,
wherein the multivalent anions as stabilizing agent have a charge of minus 2.
9. An aqueous solution composition according to any one of claims 1 to 7,
wherein the multivalent anions as stabilizing agent have a charge of minus 3.
30
10. An aqueous solution composition according to any one of claims 1 to 7,
wherein the multivalent anions are a mixture of anions having charge of minus 2
and anions having charge of minus 3.
5 11. An aqueous solution composition according to any one of claims 1 to 7,
wherein the multivalent anions having a charge of at least minus 2 are selected
from divalent citrate anions, trivalent citrate anions, divalent sulphate anions,
divalent phosphate anions, trivalent phosphate anions and mixtures thereof.
10 12. An aqueous solution composition according to any one of claims 1 to 11,
wherein the multivalent anions are a mixture of at least two different multivalent
anions having a charge of at least minus 2.
13. An aqueous solution composition according to claim 11, wherein the
15 multivalent anions having a charge of at least minus 2 are selected from divalent
citrate anions, trivalent citrate anions and mixtures thereof.
14. An aqueous solution composition according to any one of claims 1 to 13,
wherein the total concentration of multivalent anions in the composition having a
20 charge of at least minus 2 is at least 20 mM, such as at least 30 mM, at least 40
mM or at least 50 mM.
15. An aqueous solution composition according to any one of claims 1 to 14,
which additionally comprises a preservative such as a phenolic or benzylic
25 preservative.
16. An aqueous solution composition according to claim 15, wherein the
preservative is selected from the group consisting of phenol, m-cresol,
chlorocresol, chlorophenol, benzyl alcohol, propyl paraben, methyl paraben,
30 benzalkonium chloride and benzethonium chloride.
17. An aqueous solution composition according to claim 16, wherein the
concentration of preservative is 10-100 mM, for example 20-80 mM, such as 25-
50 mM.
35
18. An aqueous solution composition according to any one of claims 1 to 17,
further comprising a surfactant.
19. An aqueous solution composition according to claim 18, wherein the
40 surfactant is a non-ionic surfactant such as a polysorbate.
20. An aqueous solution composition according to claim 18, wherein the
surfactant is a cationic surfactant such as benzethonium salt or benzalkonium
salt.
31
21. An aqueous solution composition according to any one of claims 18 to 20,
wherein the concentration of surfactant is 1-2000 µg/ml, e.g. 5-1000 µg/ml, e.g.
10-500 µg/ml, such as 10-200 µg/ml.
5 22. An aqueous solution composition according to any one of claims 1 to 21,
further comprising a tonicity modifier.
23. An aqueous solution composition according to claim 22, wherein the
tonicity modifier is an uncharged tonicity modifier and is suitably selected from
10 glycerol, mannitol, propylene glycol, trehalose, PEG300 and PEG400.
24. An aqueous solution composition according to claim 23, wherein the
concentration of uncharged tonicity modifier is 50-1000 mM, for example 100-500
mM, such as about 300 mM.
15
25. An aqueous solution composition according to claim 22, wherein the
tonicity modifier is a charged tonicity modifier and is suitably selected from
sodium chloride, sodium sulphate, sodium acetate, sodium lactate, and amino
acids such as glycine or arginine.
20
26. An aqueous solution composition according to claim 25, wherein the
concentration of charged tonicity modifier is 25-500 mM, for example 50-250 mM
such as about 150 mM.
25 27. An aqueous solution composition according to any one of claims 1 to 24,
wherein the composition does not contain an amino acid.
28. An aqueous solution composition according to any one of claims 1 to 27,
wherein the composition comprises an additional active ingredient, for example
30 selected from glucagon, peptide YY, and a long acting insulin such as insulin
glargine or insulin degludec.
29. An aqueous solution composition according to any one of claims 1 to 28,
which is a therapeutic composition.
35
30. An aqueous solution composition according to any one of claims 1 to 29,
for use in the treatment or prevention of a disease or disorder caused by,
associated with and/or accompanied by disturbances in carbohydrate and/or lipid
metabolism.
40
31. A method of treating or preventing a disease or disorder caused by,
associated with and/or accompanied by disturbances in carbohydrate and/or lipid
metabolism comprising administering to a subject in need thereof a
32
therapeutically effective amount of an aqueous solution composition according to
any one of claims 1 to 29.
32. The aqueous solution composition for use according to claim 30, or the
5 method according to claim 31, wherein the disease or disorder is selected from
type 1 diabetes, type 2 diabetes, hyperglycaemia, impaired glucose tolerance,
obesity and metabolic syndrome.
33. Use of multivalent anions having a charge of at least minus 2 for
10 stabilizing an aqueous solution composition of a GLP-1 receptor agonist.
34. A method of improving the stability of an aqueous solution composition
comprising a GLP-1 receptor agonist as active ingredient, which comprises
adding multivalent anions having a charge of at least minus 2 to the composition.
15
35. A container containing one dose or a plurality of doses of an aqueous
solution composition according to any one of claims 1 to 29.
36. A container according to claim 35, which is a vial.
20
37. An injection device for single or multiple-use comprising a container
according to claim 35 together with an injection needle.
38. An injection device according to claim 37, in the form of a pen.
25
39. An injection device according to claim 37, in the form of a pump.