U.S. patent application number 17/416661 was filed with the patent office on 2022-03-10 for novel composition.
The applicant listed for this patent is ARECOR LIMITED. Invention is credited to David GERRING, Sarah HOWELL, Jan JEZEK.
Application Number | 20220072090 17/416661 |
Document ID | / |
Family ID | 69104798 |
Filed Date | 2022-03-10 |
United States Patent
Application |
20220072090 |
Kind Code |
A1 |
JEZEK; Jan ; et al. |
March 10, 2022 |
NOVEL COMPOSITION
Abstract
There is provided inter alia an aqueous solution composition of
pH in the range 5.0 to 8.0 comprising: --daptomycin or an analogue
thereof, or a salt thereof; --a divalent metal cation; and
--optionally one or more buffers being substances having at least
one ionisable group with a pK.sub.a in the range 3.0 to 9.0 and
which pK.sub.a is within 2 pH units of the pH of the composition;
wherein the buffers are present at a total concentration of 0-5
mM.
Inventors: |
JEZEK; Jan; (Saffron Walden,
GB) ; GERRING; David; (Saffron Walden, GB) ;
HOWELL; Sarah; (Saffron Walden, GB) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
ARECOR LIMITED |
Saffron Walden |
|
GB |
|
|
Family ID: |
69104798 |
Appl. No.: |
17/416661 |
Filed: |
December 20, 2019 |
PCT Filed: |
December 20, 2019 |
PCT NO: |
PCT/GB2019/053655 |
371 Date: |
June 21, 2021 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 38/063 20130101;
A61K 47/183 20130101; A61K 47/10 20130101; A61K 31/198 20130101;
A61K 47/38 20130101; A61K 9/0019 20130101; A61K 9/08 20130101; A61K
38/12 20130101; A61K 47/22 20130101; A61K 47/12 20130101; A61K
47/02 20130101; A61P 31/04 20180101; A61K 31/198 20130101; A61K
2300/00 20130101; A61K 38/063 20130101; A61K 2300/00 20130101; A61K
38/12 20130101; A61K 2300/00 20130101 |
International
Class: |
A61K 38/12 20060101
A61K038/12; A61K 9/08 20060101 A61K009/08; A61K 47/02 20060101
A61K047/02; A61K 47/22 20060101 A61K047/22; A61K 47/10 20060101
A61K047/10; A61K 47/18 20060101 A61K047/18 |
Foreign Application Data
Date |
Code |
Application Number |
Dec 21, 2018 |
GB |
1821149.0 |
Aug 13, 2019 |
GB |
1911581.5 |
Claims
1-35. (canceled)
36. An aqueous solution composition of pH in the range 5.0 to 8.0
comprising: a. daptomycin or an analogue thereof, or a salt
thereof, and b. a divalent metal cation.
37. The aqueous solution composition of claim 36, wherein the
concentration of divalent metal cation is 10-150 mM.
38. The aqueous solution composition of claim 36, wherein the
divalent metal cation is calcium ion or a magnesium ion.
39. The aqueous solution composition of claim 36, wherein the
aqueous solution composition is substantially free of buffer.
40. The aqueous solution composition of claim 36, further
comprising a stabilizer in a stabilizing amount.
41. The aqueous solution composition of claim 36, further
comprising an antioxidant.
42. The aqueous solution composition of claim 41, the antioxidant
is selected from the group consisting of methionine, glutathione,
ascorbate, butylated hydroquinone, lactate, nicotinamide,
nicotinate, tryptophan, phenylalanine and tyrosine.
43. The aqueous solution composition of claim 36, further
comprising a tonicity modifier.
44. The aqueous solution composition of claim 43, wherein the
tonicity modifier is an uncharged tonicity modifier and is selected
from the group consisting of sugars, sugar alcohols, other polyols
and polyethylene glycols.
45. The aqueous solution composition of claim 44, wherein the
uncharged tonicity modifier is selected from the group consisting
of glycerol, 1,2-propanediol, mannitol, sorbitol, sucrose,
trehalose, lactose, PEG300 and PEG400.
46. The aqueous solution composition of claim 44, wherein the
concentration of the uncharged tonicity modifier is 50-5000 mM.
47. The aqueous solution composition of claim 43, wherein the
tonicity modifier is a charged tonicity modifier, the charged
tonicity modifier is selected from the group consisting of sodium
chloride, sodium sulphate, and an amino acid.
48. The aqueous solution composition of claim 36, further
comprising one or more buffers being substances having at least one
ionisable group with a pKa in the range 3.0 to 9.0 and which pKa is
within 2 pH units of the pH of the composition, wherein the buffers
are present at a total concentration of 0-5 mM.
49. The aqueous solution composition of claim 48, wherein the one
or more buffers is selected from the group consisting of maleate,
sulphite, aspartame, aspartate, glutamate, tartrate, gluconate,
adenine, succinate, ascorbate, benzoate, phenylacetate, gallate,
cytosine, histidine, p-aminobenzoic acid, sorbate, acetate,
propionate, alginate, urate, 2-(N-morpholino)ethanesulphonic acid,
bicarbonate, bis(2-hydroxyethyl) iminotris(hydroxymethyl)methane,
A/-(2-acetamido)-2-iminodiacetic acid,
2-[(2-amino-2-oxoethyl)amino]ethanesulphonic acid,
piperazine-N,N'-bis(2-ethanesulphonic acid) (PIPES), phosphate,
A/,/V-bis(2-hydroxyethyl)-2-aminoethanesulphonic acid,
3-[A/,A/-bis(2-hydroxyethyl)amino]-2-hydroxypropanesulphonic acid,
triethanolamine, piperazine-A/.A/'-bis(2-hydroxypropanesulphonic
acid),
tris(hydroxymethyl)aminomethane,A/-tris(hydroxymethyl)glycine,
A/-tris(hydroxymethyl)methyl-3-am inopropanesulphonic acid, citrate
and salts thereof, and combinations thereof.
50. The aqueous solution composition of claim 36, wherein
composition comprises no more than 5% total impurities by total
weight of the daptomycin or an analogue thereof, or a salt thereof
in the aqueous solution composition, as measured by RP-HPLC,
following storage at 2-8.degree. C. for at least 6 months.
51. The aqueous solution composition of claim 36, wherein the
composition comprises no more than 5% total impurities by total
weight of daptomycin or an analogue thereof, or a salt thereof in
the aqueous solution composition, as measured by RP-HPLC, following
storage at 25.degree. C. for at least 2 weeks.
52. An aqueous solution composition of pH in the range 5.0 to 8.0
comprising: a. daptomycin or an analogue thereof, or a salt
thereof; b. a divalent metal cation; and c. a tonicity modifier, an
amino acid, or a combination thereof.
53. The aqueous solution composition of claim 52, wherein the
tonicity modifier is an uncharged tonicity modifier and is selected
from the group consisting of sugars, sugar alcohols, other polyols
and polyethylene glycols.
54. The aqueous solution composition of claim 52, wherein the
tonicity modifier is a charged tonicity modifier, the charged
tonicity modifier is selected from the group consisting of sodium
chloride, sodium sulphate, and an amino acid.
55. The aqueous solution composition of claim 52, wherein the amino
acid is selected from the group consisting of methionine, glycine,
proline, arginine, lysine, aspartic acid and glutamic acid.
56. The aqueous solution composition of claim 52, further
comprising one or more buffers being substances having at least one
ionisable group with a pKa in the range 3.0 to 9.0 and which pKa is
within 2 pH units of the pH of the composition, wherein the buffers
are present at a total concentration of 0-5 mM.
Description
[0001] This invention relates to aqueous solution compositions of
daptomycin, in particular at low buffer concentrations.
BACKGROUND
[0002] Daptomycin is a cyclic lipopeptide antibacterial agent
derived from the fermentation of Streptomyces roseosporus and is
produced by Merck & Co., Inc. as an antibacterial agent for use
in humans under the brand name of Cubicin.RTM.. Cubicin.RTM. is
supplied in a single-use vial as a sterile, preservative-free,
lyophilized cake containing 500 mg of daptomycin for intravenous
(IV) use following reconstitution with 0.9% sodium chloride
injection. Cubicin.RTM. is indicated for the treatment of the
complicated skin and skin structure infections (cSSSI), and for the
treatment of Staphylococcus Aureus bloodstream infections
(bacteremia).
[0003] Cubicin.RTM. should be administered intravenously either by
injection over a two minute period or by infusion over a thirty
minute period. The treatment regimen is typically once every 24
hours for 1 to 6 weeks, depending on the indication.
[0004] Lyophilized daptomycin is reconstituted shortly before use
because in currently available formulations, once reconstituted,
the daptomycin will degrade via, inter alia, hydrolysis. The need
to reconstitute the lyophilised preparation of daptomycin prior to
the IV injection or IV infusion is a considerable complication of
the administration procedure. Therefore, developing a stable liquid
composition of daptomycin is very desirable, potentially leading to
a significant simplification of the administration as well as
improving the safety of the delivery due to reduced likelihood of
error during the complex procedure for reconstitution and
preparation of the solution for infusion.
[0005] The liquid formulation of daptomycin must meet a number of
criteria in order to be approved for use in humans, including:
[0006] good physical stability (i.e. minimal aggregation and no gel
formation during storage and use) [0007] good chemical stability
(i.e. impurity levels within specified limits) [0008] minimal or no
injection site reaction (e.g. pain on injection, reddening of the
injection site) [0009] acceptable safety profile.
[0010] The objective of the present invention is the provision of a
liquid formulation of daptomycin, particularly at high strength
such as at 50 mg/ml, which has good physical and chemical stability
and is suitable for intravenous use in humans.
[0011] In particular, the objective of the present invention is the
provision of a stable liquid formulation of daptomycin (for
example, at 50 mg/ml) that is suitable for an IV delivery (either
as an IV injection or as an IV infusion).
[0012] WO2011/035108A1 and WO2011/062676A1 (both EAGLE
PHARMACEUTICALS, INC.) disclose compositions containing daptomycin
which are said to have long term storage stability.
[0013] WO2018/073269A1 (XELLIA PHARMACEUTICALS APS) discloses
formulations comprising daptomycin, one or more polar protic
solvents, and mixtures thereof.
[0014] US2002/0111311A1 (GOVARDHAN ET AL.) discloses methods of
preparing daptomycin in crystalline form.
SUMMARY OF THE INVENTION
[0015] Thus, according to the present invention, there is provided
an aqueous solution composition of pH in the range 5.0 to 8.0
comprising: [0016] daptomycin or an analogue thereof, or a salt
thereof; and [0017] a divalent metal cation; and [0018] optionally
one or more buffers being substances having at least one ionisable
group with a pK.sub.a in the range 3.0 to 9.0 and which pK.sub.a is
within 2 pH units of the pH of the composition;
[0019] wherein the buffers are present at a total concentration of
0-5 mM.
DETAILED DESCRIPTION OF THE INVENTION
[0020] Described herein are stable aqueous solution compositions of
daptomycin and a divalent cation, wherein the compositions have no
or very low concentration of buffer.
[0021] Daptomycin is a cyclic lipopeptide antibacterial agent with
chemical name
N-decanoyl-L-tryptophyl-D-asparaginyl-L-aspartyl-Lthreonylglycyl-L-ornith-
yl-L-aspartyl-D-alanyl-L-aspartylglycyl-D-seryl-threo-3-methyl-L-glutamyl--
3-anthraniloyl-L-alanine .epsilon.1-lactone. Daptomycin is also
known as LY 146032, and is a member of the factor A-219780.sub.0
type antibacterial agents of Streptomyces roseosporus. The chemical
structure of daptomycin is:
##STR00001##
[0022] The empirical formula of daptomycin is
C.sub.72H.sub.10N.sub.17O.sub.26 and its molecular weight is
1619.71.
[0023] Analogues of daptomycin include A-219780.sub.0 type
antibacterial agents.
[0024] Salts of daptomycin and its analogues thereof may be formed
with a suitable counter ion including, but not limited to, sodium,
potassium, magnesium, calcium, manganese and zinc.
[0025] Compositions containing a higher concentration of daptomycin
are of particular interest. Formulating aqueous peptide
therapeutics at higher concentrations is challenging due to
increased rate of molecular interactions between the peptide
molecules. The increased molecular interactions can lead to greater
rate of aggregation, fibril formation as well as other degradation
pathways that are due to inter-molecular interactions.
[0026] In one embodiment, the concentration of daptomycin in the
composition is >25 mg/ml, such as .gtoreq.27 mg/ml, .gtoreq.30
mg/ml, .gtoreq.35 mg/ml, .gtoreq.40 mg/ml, .gtoreq.45 mg/ml or
.gtoreq.50 mg/ml and e.g. up to around 100 mg/ml. In one
embodiment, the concentration of daptomycin in the composition is
30-100 mg/ml such as 30-60 mg/ml, such as 35-60 mg/ml, 40-60 mg/ml
or 45-55 mg/ml. In one embodiment, the concentration of daptomycin
in the composition is about 50 mg/ml.
[0027] The term "aqueous solution composition", as used herein,
refers to a solution composition in water, preferably distilled
water, deionized water, water for injection, sterile water for
injection or bacteriostatic water for injection.
[0028] The composition of the invention comprises water in an
amount of at least 65% (w/v), such as at least 70% (w/v), at least
75% (w/v), at least 80% (w/v), at least 85% (w/v), or at least 90%
(w/v).
[0029] In one embodiment, the composition solvent comprises (based
on volume) at least 80% water, such as at least 85% water, at least
90% water, at least 95% water or at least 99% water. In one
embodiment, the solvent is water i.e. the only solvent present in
the composition is water.
[0030] The present inventors have discovered that the adjustment of
certain composition parameters and the addition of certain
additives to the composition, can provide daptomycin compositions
with improved chemical and/or physical stability.
[0031] Firstly, the present inventors have discovered that in
certain damptomycin compositions, the presence of a divalent metal
ion can improve the stability of the daptomycin. As can be seen in
Example 1, both calcium chloride and magnesium chloride improved
the stability of daptomycin, with calcium chloride showing the
largest effect.
[0032] In one embodiment, the divalent metal cation is selected
from calcium, magnesium, manganese and zinc ions. In one
embodiment, the divalent metal cation is calcium ion. In another
embodiment, the divalent metal cation is magnesium ion.
[0033] The divalent metal cation will typically be added to the
composition in the form of a salt e.g. as a chloride salt, a
nitrate salt, an acetate salt, a lactate salt or a sulfate salt.
The divalent metal cation can also be added to the composition as a
hydroxide.
[0034] In one embodiment, the concentration of divalent metal
cation in the composition is 10-150 mM, such as 20-130 mM, 30-120
mM, 40-100 mM, 40-80 mM or 50-70 mM.
[0035] The pH of the composition is between 5.0 and 8.0 and can be,
for example, between 5.5 and 7.5. As can be seen from Examples 1
and 2, the optimal pH for daptomycin in the presence of calcium
ions for the compositions tested was 6.0-7.0. Thus, in a preferred
embodiment, the pH of the composition is in the range 6.0 to 7.0.
All references herein to "pH" refer to the pH of a composition
evaluated at 25.degree. C. When the pH of the composition is in the
range 6.0 to 7.0, the composition optionally comprises one or more
buffers being substances having at least one ionisable group with a
pK.sub.a in the range 4.0 to 9.0 and which pK.sub.a is within 2 pH
units of the pH of the composition.
[0036] The present inventors have also discovered that in certain
daptomycin compositions, the presence of a buffer has a detrimental
impact on the stability of the daptomycin, in a
concentration-dependent manner. As can be seen in Example 1, the
presence of 50 mM of buffer (such as phosphate and ADA) resulted in
an impairment in stability of the compositions tested. Example 4
shows that histidine buffer can be deleterious to the stability of
the formulation at concentrations of 10 mM or more. Example 4 also
shows that ADA buffer can cause instability even at 20 mM. Thus,
the concentration of buffer in the composition should be limited as
much as possible.
[0037] Without wishing to be bound by theory, it is thought that
daptomycin, when formulated at the concentrations and pH ranges
described herein, is itself a relatively good buffer. This
self-buffering capacity of daptomycin means that the addition of a
buffer to the composition is not essential to ensure sufficient
buffering capacity, and the present inventors have discovered that
the addition of further buffer to the composition can actually be
destabilizing. It should be noted that the "total concentration" of
buffers in the claimed composition excludes the concentration of
daptomycin in the composition. However, when calculating the
concentration of buffer in the aqueous solution composition, any
counterions in salt forms of daptomycin with buffering capacity
should be included in the total concentration of buffer.
[0038] In one embodiment, the total concentration of buffers in the
solution is 0-5 mM, such as 0-4 mM, 0-3 mM, 0-2 mM, 0-1 mM, 0-0.5
mM, 0-0.4 mM, 0-0.3 mM, 0-0.2 mM or 0-0.1 mM. In one embodiment,
the total concentration of buffers in the solution is 0-4 mM, such
as 0-3 mM, 0-2 mM, 0-1 mM, 0-0.5 mM, 0-0.4 mM, 0-0.3 mM, 0-0.2 mM
or 0-0.1 mM. In one embodiment, the total concentration of buffers
in the solution is 0-3 mM, such as 0-2 mM, 0-1 mM, 0-0.5 mM, 0-0.4
mM, 0-0.3 mM, 0-0.2 mM or 0-0.1 mM. In one embodiment, the total
concentration of buffers in the solution is 0-2 mM, such as 0-1 mM,
0-0.5 mM, 0-0.4 mM, 0-0.3 mM, 0-0.2 mM or 0-0.1 mM. In one
embodiment, the total concentration of buffers in the solution is
0-1 mM, such as 0-0.5 mM, 0-0.4 mM, 0-0.3 mM, 0-0.2 mM or 0-0.1
mM.
[0039] In one embodiment, the total concentration of buffers in the
solution is 0.1-5 mM, such as 0.1-4 mM, 0.1-3 mM, 0.1-2 mM, 0.1-1
mM, 0.1-0.5 mM, 0.1-0.4 mM, 0.1-0.3 mM or 0.1-0.2 mM. In one
embodiment, the total concentration of buffers in the solution is
0.1-4 mM, such as 0.1-3 mM, 0.1-2 mM, 0.1-1 mM, 0.1-0.5 mM, 0.1-0.4
mM, 0.1-0.3 mM or 0.1-0.2 mM. In one embodiment, the total
concentration of buffers in the solution is 0.1-3 mM, such as 0.1-2
mM, 0.1-1 mM, 0.1-0.5 mM, 0.1-0.4 mM, 0.1-0.3 mM or 0.1-0.2 mM. In
one embodiment, the total concentration of buffers in the solution
is 0.1-2 mM, such as 0.1-1 mM, 0.1-0.5 mM, 0.1-0.4 mM, 0.1-0.3 mM
or 0.1-0.2 mM. In one embodiment, the total concentration of
buffers in the solution is 0.1-1 mM, such as 0.1-0.5 mM, 0.1-0.4
mM, 0.1-0.3 mM or 0.1-0.2 mM.
[0040] In one embodiment, the total concentration of buffers in the
solution is 0.2-5 mM such as 0.2-4 mM, 0.2-3 mM, 0.2-2 mM, 0.2-1
mM, 0.2-0.5 mM, 0.2-0.4 mM or 0.2-0.3 mM. In one embodiment, the
total concentration of buffers in the solution is 0.2-4 mM, such as
0.2-3 mM, 0.2-2 mM, 0.2-1 mM, 0.2-0.5 mM, 0.2-0.4 mM or 0.2-0.3 mM.
In one embodiment, the total concentration of buffers in the
solution is 0.2-3 mM, such as 0.2-2 mM, 0.2-1 mM, 0.2-0.5 mM,
0.2-0.4 mM or 0.2-0.3 mM. In one embodiment, the total
concentration of buffers in the solution is 0.2-2 mM, such as 0.2-1
mM, 0.2-0.5 mM, 0.2-0.4 mM or 0.2-0.3 mM. In one embodiment, the
total concentration of buffers in the solution is 0.2-1 mM, such as
0.2-0.5 mM, 0.2-0.4 mM or 0.2-0.3 mM.
[0041] In one embodiment, the total concentration of buffers in the
solution is 0.3-5 mM, such as 0.3-4 mM, 0.3-3 mM, 0.3-2 mM, 0.3-1
mM, 0.3-0.5 mM or 0.3-0.4 mM. In one embodiment, the total
concentration of buffers in the solution is 0.3-4 mM, such as 0.3-3
mM, 0.3-2 mM, 0.3-1 mM, 0.3-0.5 mM or 0.3-0.4 mM. In one
embodiment, the total concentration of buffers in the solution is
0.3-3 mM, such as 0.3-2 mM, 0.3-1 mM, 0.3-0.5 mM or 0.3-0.4 mM. In
one embodiment, the total concentration of buffers in the solution
is 0.3-2 mM, such as 0.3-1 mM, 0.3-0.5 mM or 0.3-0.4 mM. In one
embodiment, the total concentration of buffers in the solution is
0.3-1 mM, such as 0.3-0.5 mM or 0.3-0.4 mM.
[0042] In one embodiment, the total concentration of buffers in the
solution is 0.4-5 mM, such as 0.4-4 mM, 0.4-3 mM, 0.4-2 mM, 0.4-1
mM or 0.4-0.5 mM. In one embodiment, the total concentration of
buffers in the solution is 0.4-4 mM such as 0.4-3 mM, 0.4-2 mM,
0.4-1 mM or 0.4-0.5 mM. In one embodiment, the total concentration
of buffers in the solution is 0.4-3 mM, such as 0.4-2 mM, 0.4-1 mM
or 0.4-0.5 mM. In one embodiment, the total concentration of
buffers in the solution is 0.4-2 mM, such as 0.4-1 mM or 0.4-0.5
mM. In one embodiment, the total concentration of buffers in the
solution is 0.4-1 mM or 0.4-0.5 mM.
[0043] In one embodiment, the total concentration of buffers in the
solution is 0.5-5 mM, such as 0.5-4 mM, 0.5-3 mM, 0.5-2 mM or 0.5-1
mM. In one embodiment, the total concentration of buffers in the
solution is 0.5-4 mM such as 0.5-3 mM, 0.5-2 mM or 0.5-1 mM. In one
embodiment, the total concentration of buffers in the solution is
0.5-3 mM, such as 0.5-2 mM or 0.5-1 mM. In one embodiment, the
total concentration of buffers in the solution is 0.5-2 mM or 0.5-1
mM.
[0044] In one embodiment, the total concentration of buffers in the
composition is .ltoreq.5 mM, such as .ltoreq.4 mM, .ltoreq.3 mM,
.ltoreq.2 mM, .ltoreq.1 mM, .ltoreq.0.5 mM, 0.4 mM, 0.3 mM,
.ltoreq.0.2 mM or .ltoreq.0.1 mM.
[0045] In one embodiment, the composition is substantially free of
buffer. As used herein, "substantially free" means the total
concentration of buffers in the aqueous solution composition is
less than 0.1 mM. To reiterate, when considering the concentration
of buffer in the solution composition, any buffering capacity of
the daptomycin itself should be excluded. In one embodiment, the
composition does not contain buffer.
[0046] The buffer(s) where present will have buffering capacity at
the pH of the composition. Buffers typically comprise ionisable
groups with pK.sub.a within 1 pH unit of the pH of the composition,
however, a moiety which has ionisable groups with pK.sub.a more
than 1 pH unit greater or less than the pH of the composition may
also provide some buffering effect if present in a sufficient
amount. In one embodiment, the (or a) buffer comprises ionisable
groups with pK.sub.a within 1 pH unit of the pH of the composition.
In another embodiment, the (or a) buffer comprises ionisable groups
with pK.sub.a within 1.5 pH units of the pH of the composition
(such as between 1 and 1.5 pH units of the pH of the composition).
In a further embodiment, the (or a) buffer comprises ionisable
groups with pK.sub.a within 2 pH units of the pH of the composition
(such as between 1.5 and 2 pH units of the pH of the composition).
Buffering capacity is suitably determined at 25.degree. C.
[0047] Compositions which are substantially free of buffers are
suitably substantially free of substances having at least one
ionisable group with a pK.sub.a which is within 2 pH units of the
pH of the composition.
[0048] In an embodiment, the composition contains a single buffer.
In an embodiment, the composition contains two buffers.
[0049] In one embodiment, the composition contains a buffer with a
single ionisable group. In one embodiment, the composition does not
contain zwitterionic species.
[0050] The pH of an aqueous solution decreases if an acid is added
and increases if a base is added. At a given temperature and
atmospheric pressure, the magnitude of the pH decrease on addition
of an acid or the magnitude of the pH increase on addition of a
base depends on (1) the amount of the acid or the base added, (2)
the starting pH of the aqueous solution (i.e. prior to the addition
of the acid or the base) and (3) the presence of a buffer. Thus,
(1) starting from a given pH, the addition of a greater amount of
an acid or a base will result in greater magnitude of pH change,
(2) addition of a given amount of an acid or a base will result in
the greatest pH change at neutral pH (i.e. pH 7.0) and the
magnitude of the pH change will decrease as the starting pH moves
away from pH 7.0 and (3) the magnitude of the pH change, starting
from a given pH, will be smaller in the presence of a buffer than
in the absence of a buffer. A buffer thus has the ability to reduce
the change in pH if an acid or a base is added to the solution.
[0051] Suitably, a substance is considered to be a buffer if it is
capable of reducing the magnitude of the pH change of a solution to
75%, preferably 50%, most preferably to 25%, compared with an
identical solution that does not comprise the buffer, when either
strong acid or a strong base is added resulting in 0.1 mM increase
of the acid or the base in the solution.
[0052] Conversely, suitably, a substance is not considered to be a
buffer if it is not capable of reducing the magnitude of the pH
change of a solution to 75%, preferably 50%, most preferably to
25%, compared with an identical solution that does not comprise the
substance, when either strong acid or a strong base is added
resulting in 0.1 mM increase of the acid or the base in the
solution.
[0053] In one embodiment, the or a buffer is an amino acid. In
another embodiment, the or a buffer is not an amino acid.
[0054] In one embodiment, the composition comprises a buffer or
buffers selected from the group consisting of maleate, sulphite,
glyoxylate, aspartame, glucuronate, aspartate, glutamate, tartrate,
gluconate, lactate, glycolic acid, adenine, succinate, ascorbate,
benzoate, phenylacetate, gallate, cytosine, p-aminobenzoic acid,
sorbate, acetate, propionate, alginate, urate,
2-(N-morpholino)ethanesulphonic acid, bicarbonate,
bis(2-hydroxyethyl) iminotris(hydroxymethyl)methane,
N-(2-acetamido)-2-iminodiacetic acid,
2-[(2-amino-2-oxoethyl)amino]ethanesulphonic acid,
piperazine-N,N'-bis(2-ethanesulphonic acid) (PIPES), phosphate,
N,N-bis(2-hydroxyethyl)-2-aminoethanesulphonic acid,
3-[N,N-bis(2-hydroxyethyl)amino]-2-hydroxypropanesulphonic acid,
triethanolamine, piperazine-N,N'-bis(2-hydroxypropanesulphonic
acid), 2-[(2-amino-2-oxoethyl)-(carboxymethyl)amino]acetic acid
(ADA), tris(hydroxymethyl)aminomethane,
N-tris(hydroxymethyl)glycine and N-tris(hydroxymethyl)methyl-3-am
inopropanesulphonic acid, and salts thereof, and combinations
thereof. Such buffers contain ionisable groups with pK.sub.a in the
range 3.0-9.0.
[0055] In one embodiment, the composition comprises a buffer or
buffers selected from the group consisting of maleate, sulphite,
aspartame, aspartate, glutamate, tartrate, adenine, succinate,
ascorbate, benzoate, phenylacetate, gallate, cytosine,
p-aminobenzoic acid, sorbate, acetate, propionate, alginate, urate,
2-(N-morpholino)ethanesulphonic acid, bicarbonate,
bis(2-hydroxyethyl) iminotris(hydroxymethyl)methane,
N-(2-acetamido)-2-iminodiacetic acid,
2-[(2-amino-2-oxoethyl)amino]ethanesulphonic acid,
piperazine-N,N'-bis(2-ethanesulphonic acid) (PIPES), phosphate,
N,N-bis(2-hydroxyethyl)-2-aminoethanesulphonic acid,
3-[N,N-bis(2-hydroxyethyl)amino]-2-hydroxypropanesulphonic acid,
triethanolamine, piperazine-N,N'-bis(2-hydroxypropanesulphonic
acid), 2-[(2-amino-2-oxoethyl)-(carboxymethyl)amino]acetic acid
(ADA), tris(hydroxymethyl)aminomethane,
N-tris(hydroxymethyl)glycine, N-tris(hydroxymethyl)methyl-3-am
inopropanesulphonic acid, citrate and salts thereof, and
combinations thereof. Such buffers contain ionisable groups with
pK.sub.a in the range 4.0-9.0.
[0056] In one embodiment, the buffer is selected from the group
consisting of maleate, tartrate, lactate, benzoate, acetate,
bicarbonate, phosphate, tris(hydroxymethyl)aminomethane and
citrate, in particular lactate, acetate, phosphate,
tris(hydroxymethyl)aminomethane and citrate, especially
phosphate.
[0057] It should be noted that all references to "pK.sub.a" refer
to the pK.sub.a of an ionisable group evaluated at 25.degree. C.
(see CRC Handbook of Chemistry and Physics, 79.sup.th Edition,
1998, D. R. Lide).
[0058] The addition of one or more stabilizers to the composition
can provide further stability benefits. A stabilizer will be
employed in a stabilizing amount.
[0059] At certain pH ranges the stabilizers, or indeed any other
additives described herein (including chelating agents, amphipathic
species etc) may have buffering capacity, e.g. in a composition of
pH in the range 5.0-8.0, the stabilizer or additive may have at
least one ionisable group with a pK.sub.a in the range 3.0 to 9.0
and which pK.sub.a is within 2 pH units of the pH of the
composition. For the avoidance of doubt, the concentration of such
stabilizers or additives should be included in the total
concentration of buffers. Such stabilizers or additives are
therefore present in the composition at a concentration which is
sufficiently low such that the total buffer concentration
limitation is met e.g. does not exceed 5 mM.
[0060] In one embodiment, the stabilizer is selected from amino
acids, particularly natural amino acids, such as .alpha.-amino
acids. In one embodiment, the amino acid is selected from the group
consisting of methionine, glycine, proline, arginine, lysine,
aspartic acid and glutamic acid, and in particular is selected from
the group consisting of consisting of glycine, proline, methionine,
arginine, lysine and aspartic acid e.g. is selected from the group
consisting of consisting of glycine, proline, methionine, arginine
and lysine. Suitably, the amino acid is present at a concentration
of 1-200 mM, such as 1-100 mM, 1-50 mM, 1-20 mM, 1-10 mM, 1-5 mM,
1-4 mM, 1-3 mM or 1-2 mM.
[0061] In one embodiment, the amino acid is not a buffer at the
given pH of the composition i.e. the amino acid does not comprise
ionisable groups with pK.sub.a within 2 pH units, such as 1.5 pH
units or 1 pH unit of the pH of the composition. In one embodiment
the amino acid is not histidine.
[0062] When the pH of the composition is in the range of 5.0 to
8.0, when present, the amino acid suitably does not have side
chains with pK.sub.a in the range 3.0 to 9.0. In one embodiment, an
amino acid is selected from the group consisting of consisting of
glycine, proline, methionine, arginine and lysine.
[0063] When the pH of the composition is in the range 6.0 to 7.0,
when present, the amino acid suitably does not have side chains
with pK.sub.a in the range 4.0 to 9.0. In one embodiment, the amino
acid is suitably selected from the group consisting of glycine,
proline, methionine, arginine, lysine and aspartic acid.
[0064] In one embodiment, an amino acid is present in the
composition and is glutamic acid, at a concentration of 5 mM or
less. In another embodiment, the amino acid is present in the
composition and is selected from aspartic acid, at a concentration
of 5 mM or less.
[0065] The composition may comprise a tonicity modifier, which may
be charged or uncharged. In certain embodiments, the tonicity
modifier can have a stabilizing effect. Examples of uncharged
tonicity modifiers include sugars (such as sucrose, trehalose and
lactose), sugar alcohols (such as mannitol and sorbitol), other
polyols (such as glycerol and 1,2-propanediol) and polyethylene
glycols (such as PEG300 and PEG400). In one embodiment, the
uncharged tonicity modifier is selected from the group consisting
of glycerol, 1,2-propanediol, mannitol, sorbitol, sucrose,
trehalose, lactose, PEG300 and PEG400. When included, an uncharged
tonicity modifier is typically employed in the composition at a
concentration of 50-5000 mM, for example 100-1000 mM, such as
100-500 mM, such as 200-400 mM, such as about 300 mM. Examples of
charged tonicity modifiers include sodium chloride, sodium
sulphate, and amino acids such as glycine or arginine. In one
embodiment, the charged tonicity modifier is selected from the
group consisting of sodium chloride, sodium sulphate, and amino
acids such as glycine or arginine, and in particular selected from
sodium chloride and sodium sulphate. In one embodiment, the charged
tonicity modifier is sodium chloride. Amino acids which have
buffering capacity within the pH range of the composition are
suitably employed at a concentration which is sufficiently low such
that the total buffer concentration does not exceed 5 mM. When
included, a charged tonicity modifier such as sodium chloride is
typically employed in the composition at a concentration of 50-2000
mM, for example 50-1000 mM, such as 50-500 mM such as 100-200 mM,
such as about 150 mM; or at a concentration of 0.1-5 mM such as
0.1-4 mM, 0.1-3 mM, 0.1-2 mM or 0.1-1 mM. Alternatively, a charged
tonicity modifier such as sodium chloride may be employed in the
composition at a concentration of 300 mM or more e.g. 500 mM or
more e.g. 500-3000 mM e.g. 1000-2500 mM e.g. e.g. 1500-2500 mM e.g.
around 2000 mM. As shown in Example 2 and 3, high concentrations of
sodium chloride significantly increased the stability of the
compositions of the invention.
[0066] The composition may optionally comprise a surfactant,
suitably at a concentration of 0.01-10 mg/ml, such as 0.05-2
mg/ml.
[0067] In one embodiment, the surfactant is a non-ionic surfactant.
A particularly 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 glucoside,
hexadecyl maltoside, sucrose monooctanoate, sucrose mono decanoate,
sucrose monododecanoate, sucrose monotridecanoate, sucrose
monotetradecanoate and sucrose monohexadecanoate.
[0068] Another suitable class of non-ionic surfactants is the
polysorbates (fatty acid esters of ethoxylated sorbitan), such as
polysorbate 20 or polysorbate 80. Polysorbate 20 is a mono ester
formed from lauric acid and polyoxyethylene (20) sorbitan in which
the number 20 indicates the number of oxyethylene groups in the
molecule. Polysorbate 80 is a 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. Polysorbate 80 is known under a range of
brand names including in particular Tween 80, and also Alkest TW
80. Other suitable polysorbates include polysorbate 40 and
polysorbate 60.
[0069] 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
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).
[0070] Another suitable class of non-ionic surfactants is block
copolymers of 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.
[0071] Another suitable class of non-ionic surfactants are
alkylphenyl ethers of polyethylene glycol, especially
4-(1,1,3,3-tetramethylbutyl)phenyl-polyethylene glycol, also known
under a brand name Triton X-100.
[0072] In one embodiment, the non-ionic surfactant is selected from
the group consisting of an alkyl glycoside, a polysorbate, an alkyl
ether of polyethylene glycol, a block copolymer of polyethylene
glycol and polypropylene glycol, and an alkylphenyl ether of
polyethylene glycol. In one embodiment, the concentration of the
non-ionic surfactant in the composition is 10-10,000 .mu.g/ml, such
as 10-8,000 .mu.g/ml, 10-5,000 .mu.g/ml, 10-3,000 .mu.g/ml, 10-2000
.mu.g/ml, 50-1000 .mu.g/ml, 100-500 .mu.g/ml or about 200 .mu.g/ml.
In one embodiment, the non-ionic surfactant is an alkyl glycoside
at a concentration of 10-10,000 .mu.g/ml, such as 100-10,000
.mu.g/ml, 1,000-10,000 .mu.g/ml or 5,000-10,000 .mu.g/ml.
[0073] In one embodiment, the surfactant is a cationic surfactant.
Suitable cationic surfactants include benzalkonium and benzethonium
salts. In one embodiment, the cationic surfactant is selected from
benzethonium salts e.g. 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. In one embodiment, the concentration of the cationic
surfactant in the composition is 10-2000 .mu.g/ml, such as 50-1000
.mu.g/ml, 100-500 .mu.g/ml or about 200 .mu.g/ml.
[0074] In one embodiment, the surfactant is an anionic surfactant.
Suitable anionic surfactants include sodium dodecyl sulfate,
ammonium dodecyl sulfate, carboxylate salts (sodium or ammonium
salts of stearic or palitic acid), and ether phosphates.
[0075] In one embodiment, the surfactant is a zwitterionic
surfactant. Suitable zwitterionic surfactants include
phosphatidylcholine, phosphatidylserine, phosphatitylethanolamine
and 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate
(CHAPS).
[0076] Suitably, the surfactant is a non-ionic surfactant, as
described above.
[0077] In one embodiment, the composition further comprises a
surfactant and an uncharged tonicity modifier e.g. selected from
sugars, sugar alcohols, other polyols and polyethylene glycols. In
one embodiment, the composition further comprises a surfactant and
a charged tonicity modifier, such as sodium chloride or sodium
sulphate.
[0078] The composition may optionally comprise an antioxidant. In
one embodiment, the antioxidant is selected from the group
consisting of methionine, glutathione, ascorbate, butylated
hydroquinone, lactate, nicotinamide, nicotinate, tryptophan,
phenylalanine and tyrosine and is suitably selected from
methionine, glutathione, ascorbate and butylated hydroquinone, and
in particular is methionine. Suitably, the antioxidant is present
at a concentration of 1-100 mg/ml, such as 5-50 mg/ml. Without
being bound by theory, it is thought that the presence of an
antioxidant may improve the chemical and/or physical stability of
daptomycin, for example by eliminating free radicals that may play
a part in some of the degradation pathways. In one embodiment, the
composition comprises two antioxidants e.g. selected from the group
consisting of methionine, glutathione, ascorbate, butylated
hydroquinone, lactate, nicotinamide, nicotinate, tryptophan,
phenylalanine and tyrosine.
[0079] The composition may optionally comprise a chelating agent.
By chelating agent is meant an agent capable of complexing with
metal ions particularly ions of heavy metals such as iron or
copper. Trace amounts of these metals can accelerate chemical
degradation of daptomycin. Such metals can be introduced into the
formulation as trace contaminants of the desirable salts of
divalent metals. In one embodiment, the chelating agent is selected
from the group consisting of EDTA (ethylenediaminetetraacetic
acid), EGTA (ethylene glycol-bis(.beta.-aminoethyl
ether)-N,N,N',N'-tetraacetic acid) and citrate. In one embodiment
the chelating agent is not histidine.
[0080] In one embodiment, the chelating agent is not a buffer at
the given pH of the composition i.e. the chelating agent does not
comprise ionisable groups with pK.sub.a within 2 pH units, such as
1.5 pH units or 1 pH unit of the pH of the composition. The
chelating agent, if present, is at a significantly lower
concentration that the divalent metal cation. Suitably, the
chelating agent is present at a concentration of 0.1-20 mM, such as
0.1-15 mM, 0.1-10 mM, 0.1-5 mM or 0.1-1 mM. Without being bound by
theory, although perhaps counterintuitive to add a chelating agent
to a composition requiring the presence of a divalent metal cation,
it is thought that the presence of a chelating agent well below the
concentration of the divalent metal cation may improve the chemical
and/or physical stability of daptomycin, for example by eliminating
trace quantities of metals cations (in particular heavy metals,
e.g. Fe.sup.3+) that are known to catalyse various degradation
pathways. In one embodiment, the chelating agent is a multi-anion.
By multi-anion is meant a species which has at least two anionic
centres per molecule, at the particular pH of the solution. When
the chelating agent is a multi-anion it may be employed as a
suitable salt form (e.g. as a sodium salt), or as an acid form
which forms a multi-anion in solution.
[0081] The composition may optionally comprise a polyanion. In one
embodiment, the polyanion is selected from the group consisting of
carboxymethyl cellulose and dextran sulfate. Suitably, the
polyanion is present at a concentration of 1-10 mg/ml, such as 1-5
mg/ml.
[0082] The composition may optionally comprise a polycation. In one
embodiment, the polycation is an oligomer of ethyleneimine, in
particular an oligomer of ethyleneimine, wherein the number of
repeating units of ethyleneimine (n) in the oligomer is in the
range 2-12. In one embodiment, the polycation is an oligomer of
ethyleneimine selected from the group consisting of
diethylenetriamine, triethylenetetramine (TETA),
tetraethylenepentamine and pentaethylenehexamine. In one
embodiment, the polycation is TETA. Suitably, the polycation is
present at a concentration of 1-10 mg/ml, such as 1-5 mg/ml.
[0083] Without being bound by theory, it is thought that the
presence of polyions (i.e. polyanions and/or polycations) may
improve the physical and/or chemical stability of daptomycin, for
example by forming a weak complex and thus preventing interactions
between daptomycin molecules that lead to aggregation.
[0084] The composition may optionally comprise a charged
amphiphilic species. Such a species comprises a charged region and
a non-polar (hydrophobic) region. In one embodiment, the non-polar
region is a benzene ring. In another embodiment, the non-polar
region is an aliphatic chain of four or more carbon atoms. In one
embodiment, the charged amphiphilic species is selected from the
group consisting of benzoic acid, 2-hydroxybenzoic acid,
3-hydroxybenzoic acid, 4-hydroxybenzoic acid,
3,4,5-trihydroxybenzoic acid, 4-hydroxy-3-methoxybenzoic acid,
phthalic acid, isophthalic acid, terephthalic acid, 1-naphthoic
acid, 2-naphthoic acid, indole-3-acetic acid, phenylacetic acid,
3-phenylpropionic acid, trans-cinnamic acid, cis-cinnamic acid and
mandelic acid. In one embodiment, the charged amphiphilic species
is benzoic acid, particularly in its ionic form (benzoate ion). In
one embodiment, the charged amphiphilic species is not a buffer at
the given pH of the composition i.e. the charged amphiphilic
species does not comprise ionisable groups with pK.sub.a within 2
pH units, such as 1.5 pH units or 1 pH unit of the pH of the
composition. Suitably, the charged amphiphilic species is present
at a concentration of 1-100 mM, such as 5-20 mM; or at a
concentration of 0.1-5 mM such as 0.1-4 mM, 0.1-3 mM, 0.1-2 mM or
0.1-1 mM.
[0085] In one aspect of the invention is provided an aqueous
solution composition of pH 5.0-8.0, suitably pH 6.0-7.0, comprising
or consisting of: [0086] daptomycin or an analogue thereof, or a
salt thereof, at a concentration of 30-60 mg/ml, such as 35-60
mg/ml, 40-60 mg/ml or 45-55 mg/ml; [0087] a divalent metal cation
selected from calcium and magnesium; and [0088] optionally one or
more buffers being substances having at least one ionisable group
with a pK.sub.a in the range 3.0 to 9.0, suitably with pK.sub.a in
the range 4.0 to 9.0; and which pK.sub.a is within 2 pH units of
the pH of the composition;
[0089] wherein the buffers are present at a total concentration of
0-5 mM, such as 0-4 mM, 0-3 mM, 0-2 mM, 0-1 mM, 0-0.5 mM, 0-0.4 mM,
0-0.3 mM, 0-0.2 mM or 0-0.1 mM; or at a total concentration of
0.1-5 mM, such as 0.1-4 mM, 0.1-3 mM, 0.1-2 mM, 0.1-1 mM, 0.1-0.5
mM, 0.1-0.4 mM, 0.1-0.3 mM or 0.1-0.2 mM; or at a total
concentration of 0.2-5 mM, such as 0.2-4 mM, 0.2-3 mM, 0.2-2 mM,
0.2-1 mM, 0.2-0.5 mM, 0.2-0.4 mM or 0.2-0.3 mM; or at a total
concentration of 0.3-5 mM, such as 0.3-4 mM, 0.3-3 mM, 0.3-2 mM,
0.3-1 mM, 0.3-0.5 mM or 0.3-0.4 mM; or at a total concentration of
0.4-5 mM, such as 0.4-4 mM, 0.4-3 mM, 0.4-2 mM, 0.4-1 mM or 0.4-0.5
mM; or at a total concentration of 0.5-5 mM, such as 0.5-4 mM,
0.5-3 mM, 0.5-2 mM or 0.5-1 mM.
[0090] In one aspect of the invention is provided an aqueous
solution composition of pH 5.0-8.0, suitably pH 6.0-7.0, comprising
or consisting of: [0091] daptomycin or an analogue thereof, or a
salt thereof, at a concentration of 30-60 mg/ml, such as 35-60
mg/ml, 40-60 mg/ml or 45-55 mg/ml; [0092] a divalent metal cation
selected from calcium and magnesium; and
[0093] wherein the aqueous solution composition is substantially
free of buffers and particularly is substantially free of
substances having at least one ionisable group with a pK.sub.a in
the range 3.0 to 9.0, suitably with pK.sub.a in the range 4.0 to
9.0 and which pK.sub.a is within 2 pH units of the pH of the
composition.
[0094] In one aspect of the invention is provided an aqueous
solution composition of pH 5.0-8.0, suitably pH 6.0-7.0, comprising
or consisting of: [0095] daptomycin or an analogue thereof, or a
salt thereof, at a concentration of 30-60 mg/ml, such as 35-60
mg/ml, 40-60 mg/ml or 45-55 mg/ml; and [0096] a divalent metal
cation selected from calcium and magnesium;
[0097] wherein the aqueous solution composition is substantially
free of buffer, wherein buffer is a substance having at least one
ionisable group with a pK.sub.a in the range 3.0 to 9.0, suitably
with pK.sub.a in the range 4.0 to 9.0; and which pK.sub.a is within
2 pH units of the pH of the composition.
[0098] In one aspect of the invention is provided an aqueous
solution composition of pH 5.0-8.0, suitably pH 6.0-7.0, comprising
or consisting of: [0099] daptomycin or an analogue thereof, or a
salt thereof, at a concentration of 30-60 mg/mi, such as 35-60
mg/ml, 40-60 mg/ml or 45-55 mg/ml; [0100] a divalent metal cation
selected from calcium and magnesium; and [0101] optionally one or
more buffers being substances having at least one ionisable group
with a pK.sub.a in the range 3.0 to 9.0, suitably with pK.sub.a in
the range 4.0 to 9.0; and which pK.sub.a is within 2 pH units of
the pH of the composition;
[0102] wherein the total concentration of buffers in the
composition is .ltoreq.1.1 mM.
[0103] In one aspect of the invention is provided an aqueous
solution composition of pH 5.0-8.0, suitably pH 6.0-7.0, comprising
or consisting of: [0104] daptomycin or an analogue thereof, or a
salt thereof, at a concentration of 30-60 mg/ml, such as 35-60
mg/ml, 40-60 mg/ml or 45-55 mg/ml; [0105] a divalent metal cation
selected from calcium and magnesium; [0106] a surfactant, suitably
a non-ionic surfactant; and [0107] optionally one or more buffers
being substances having at least one ionisable group with a
pK.sub.a in the range 3.0 to 9.0, suitably with pK.sub.a in the
range 4.0 to 9.0; and which pK.sub.a is within 2 pH units of the pH
of the composition;
[0108] wherein the buffers are present at a total concentration of
0-5 mM, such as 0-4 mM, 0-3 mM, 0-2 mM, 0-1 mM, 0-0.5 mM, 0-0.4 mM,
0-0.3 mM, 0-0.2 mM or 0-0.1 mM; or at a total concentration of
0.1-5 mM, such as 0.1-4 mM, 0.1-3 mM, 0.1-2 mM, 0.1-1 mM, 0.1-0.5
mM, 0.1-0.4 mM, 0.1-0.3 mM or 0.1-0.2 mM; or at a total
concentration of 0.2-5 mM, such as 0.2-4 mM, 0.2-3 mM, 0.2-2 mM,
0.2-1 mM, 0.2-0.5 mM, 0.2-0.4 mM or 0.2-0.3 mM; or at a total
concentration of 0.3-5 mM, such as 0.3-4 mM, 0.3-3 mM, 0.3-2 mM,
0.3-1 mM, 0.3-0.5 mM or 0.3-0.4 mM; or at a total concentration of
0.4-5 mM, such as 0.4-4 mM, 0.4-3 mM, 0.4-2 mM, 0.4-1 mM or 0.4-0.5
mM; or at a total concentration of 0.5-5 mM, such as 0.5-4 mM,
0.5-3 mM, 0.5-2 mM or 0.5-1 mM. In one embodiment, the non-ionic
surfactant is an alkylglycoside such a dodecyl maltoside, suitably
at a concentration of 10-10,000 .mu.g/ml.
[0109] In one aspect of the invention is provided an aqueous
solution composition of pH 5.0-8.0, suitably pH 6.0-7.0, comprising
or consisting of: [0110] daptomycin or an analogue thereof, or a
salt thereof, at a concentration of 30-60 mg/ml, such as 35-60
mg/ml, 40-60 mg/ml or 45-55 mg/ml; [0111] a divalent metal cation
selected from calcium and magnesium; [0112] an antioxidant,
suitably methionine; and [0113] optionally one or more buffers
being substances having at least one ionisable group with a
pK.sub.a in the range 3.0 to 9.0, suitably with pK.sub.a in the
range 4.0 to 9.0; and which pK.sub.a is within 2 pH units of the pH
of the composition;
[0114] wherein the buffers are present at a total concentration of
0-5 mM, such as 0-4 mM, 0-3 mM, 0-2 mM, 0-1 mM, 0-0.5 mM, 0-0.4 mM,
0-0.3 mM, 0-0.2 mM or 0-0.1 mM; or at a total concentration of
0.1-5 mM, such as 0.1-4 mM, 0.1-3 mM, 0.1-2 mM, 0.1-1 mM, 0.1-0.5
mM, 0.1-0.4 mM, 0.1-0.3 mM or 0.1-0.2 mM; or at a total
concentration of 0.2-5 mM, such as 0.2-4 mM, 0.2-3 mM, 0.2-2 mM,
0.2-1 mM, 0.2-0.5 mM, 0.2-0.4 mM or 0.2-0.3 mM; or at a total
concentration of 0.3-5 mM, such as 0.3-4 mM, 0.3-3 mM, 0.3-2 mM,
0.3-1 mM, 0.3-0.5 mM or 0.3-0.4 mM; or at a total concentration of
0.4-5 mM, such as 0.4-4 mM, 0.4-3 mM, 0.4-2 mM, 0.4-1 mM or 0.4-0.5
mM; or at a total concentration of 0.5-5 mM, such as 0.5-4 mM,
0.5-3 mM, 0.5-2 mM or 0.5-1 mM.
[0115] In one aspect of the invention is provided an aqueous
solution composition of pH 5.0-8.0, suitably pH 6.0-7.0, comprising
or consisting of: [0116] daptomycin or an analogue thereof, or a
salt thereof, at a concentration of 30-60 mg/ml, such as 35-60
mg/ml, 40-60 mg/ml or 45-55 mg/ml; [0117] a divalent metal cation
selected from calcium and magnesium; [0118] a chelating agent,
suitably selected from EDTA, EGTA and citrate; and [0119]
optionally one or more buffers being substances having at least one
ionisable group with a pK.sub.a in the range 3.0 to 9.0, suitably
with pK.sub.a in the range 4.0 to 9.0; and which pK.sub.a is within
2 pH units of the pH of the composition;
[0120] wherein the buffers are present at a total concentration of
0-5 mM, such as 0-4 mM, 0-3 mM, 0-2 mM, 0-1 mM, 0-0.5 mM, 0-0.4 mM,
0-0.3 mM, 0-0.2 mM or 0-0.1 mM; or at a total concentration of
0.1-5 mM, such as 0.1-4 mM, 0.1-3 mM, 0.1-2 mM, 0.1-1 mM, 0.1-0.5
mM, 0.1-0.4 mM, 0.1-0.3 mM or 0.1-0.2 mM; or at a total
concentration of 0.2-5 mM, such as 0.2-4 mM, 0.2-3 mM, 0.2-2 mM,
0.2-1 mM, 0.2-0.5 mM, 0.2-0.4 mM or 0.2-0.3 mM; or at a total
concentration of 0.3-5 mM, such as 0.3-4 mM, 0.3-3 mM, 0.3-2 mM,
0.3-1 mM, 0.3-0.5 mM or 0.3-0.4 mM; or at a total concentration of
0.4-5 mM, such as 0.4-4 mM, 0.4-3 mM, 0.4-2 mM, 0.4-1 mM or 0.4-0.5
mM; or at a total concentration of 0.5-5 mM, such as 0.5-4 mM,
0.5-3 mM, 0.5-2 mM or 0.5-1 mM.
[0121] In one aspect of the invention is provided an aqueous
solution composition of pH 5.0-8.0, suitably pH 6.0-7.0, comprising
or consisting of: [0122] daptomycin or an analogue thereof, or a
salt thereof, at a concentration of 30-60 mg/ml, such as 35-60
mg/ml, 40-60 mg/ml or 45-55 mg/ml; [0123] a divalent metal cation
selected from calcium and magnesium; [0124] a polyanion, suitably
selected from the group consisting of carboxymethyl cellulose and
dextran sulfate; and [0125] optionally one or more buffers being
substances having at least one ionisable group with a pK.sub.a in
the range 3.0 to 9.0, suitably with pK.sub.a in the range 4.0 to
9.0; and which pK.sub.a is within 2 pH units of the pH of the
composition;
[0126] wherein the buffers are present at a total concentration of
0-5 mM, such as 0-4 mM, 0-3 mM, 0-2 mM, 0-1 mM, 0-0.5 mM, 0-0.4 mM,
0-0.3 mM, 0-0.2 mM or 0-0.1 mM; or at a total concentration of
0.1-5 mM, such as 0.1-4 mM, 0.1-3 mM, 0.1-2 mM, 0.1-1 mM, 0.1-0.5
mM, 0.1-0.4 mM, 0.1-0.3 mM or 0.1-0.2 mM; or at a total
concentration of 0.2-5 mM, such as 0.2-4 mM, 0.2-3 mM, 0.2-2 mM,
0.2-1 mM, 0.2-0.5 mM, 0.2-0.4 mM or 0.2-0.3 mM; or at a total
concentration of 0.3-5 mM, such as 0.3-4 mM, 0.3-3 mM, 0.3-2 mM,
0.3-1 mM, 0.3-0.5 mM or 0.3-0.4 mM; or at a total concentration of
0.4-5 mM, such as 0.4-4 mM, 0.4-3 mM, 0.4-2 mM, 0.4-1 mM or 0.4-0.5
mM; or at a total concentration of 0.5-5 mM, such as 0.5-4 mM,
0.5-3 mM, 0.5-2 mM or 0.5-1 mM.
[0127] In one aspect of the invention is provided an aqueous
solution composition of pH 5.0-8.0, suitably pH 6.0-7.0, comprising
or consisting of: [0128] daptomycin or an analogue thereof, or a
salt thereof, at a concentration of 30-60 mg/ml, such as 35-60
mg/ml, 40-60 mg/ml or 45-55 mg/ml; [0129] a divalent metal cation
selected from calcium and magnesium; [0130] a polycation such as an
oligomer of ethyleneimine e.g. selected from the group consisting
of diethylenetriamine, triethylenetetramine (TETA),
tetraethylenepentamine and pentaethylenehexamine; and [0131]
optionally one or more buffers being substances having at least one
ionisable group with a pK.sub.a in the range 3.0 to 9.0, suitably
with pK.sub.a in the range 4.0 to 9.0; and which pK.sub.a is within
2 pH units of the pH of the composition;
[0132] wherein the buffers are present at a total concentration of
0-5 mM, such as 0-4 mM, 0-3 mM, 0-2 mM, 0-1 mM, 0-0.5 mM, 0-0.4 mM,
0-0.3 mM, 0-0.2 mM or 0-0.1 mM; or at a total concentration of
0.1-5 mM, such as 0.1-4 mM, 0.1-3 mM, 0.1-2 mM, 0.1-1 mM, 0.1-0.5
mM, 0.1-0.4 mM, 0.1-0.3 mM or 0.1-0.2 mM; or at a total
concentration of 0.2-5 mM, such as 0.2-4 mM, 0.2-3 mM, 0.2-2 mM,
0.2-1 mM, 0.2-0.5 mM, 0.2-0.4 mM or 0.2-0.3 mM; or at a total
concentration of 0.3-5 mM, such as 0.3-4 mM, 0.3-3 mM, 0.3-2 mM,
0.3-1 mM, 0.3-0.5 mM or 0.3-0.4 mM; or at a total concentration of
0.4-5 mM, such as 0.4-4 mM, 0.4-3 mM, 0.4-2 mM, 0.4-1 mM or 0.4-0.5
mM; or at a total concentration of 0.5-5 mM, such as 0.5-4 mM,
0.5-3 mM, 0.5-2 mM or 0.5-1 mM.
[0133] In one aspect of the invention is provided an aqueous
solution composition of pH 5.0-8.0, suitably pH 6.0-7.0, comprising
or consisting of: [0134] daptomycin or an analogue thereof, or a
salt thereof, at a concentration of 30-60 mg/ml, such as 35-60
mg/ml, 40-60 mg/ml or 45-55 mg/ml; [0135] a divalent metal cation
selected from calcium and magnesium; [0136] a charged amphiphilic
species suitably selected from the group consisting of benzoic
acid, 2-hydroxybenzoic acid, 3-hydroxybenzoic acid,
4-hydroxybenzoic acid, 3,4,5-trihydroxybenzoic acid,
4-hydroxy-3-methoxybenzoic acid, phthalic acid, isophthalic acid,
terephthalic acid, 1-naphthoic acid, 2-naphthoic acid,
indole-3-acetic acid, phenylacetic acid, 3-phenylpropionic acid,
trans-cinnamic acid, cis-cinnamic acid and mandelic acid; in
particular benzoic acid; and [0137] optionally one or more buffers
being substances having at least one ionisable group with a
pK.sub.a in the range 3.0 to 9.0, suitably with pK.sub.a in the
range 4.0 to 9.0; and which pK.sub.a is within 2 pH units of the pH
of the composition;
[0138] wherein the buffers are present at a total concentration of
0-5 mM, such as 0-4 mM, 0-3 mM, 0-2 mM, 0-1 mM, 0-0.5 mM, 0-0.4 mM,
0-0.3 mM, 0-0.2 mM or 0-0.1 mM; or at a total concentration of
0.1-5 mM, such as 0.1-4 mM, 0.1-3 mM, 0.1-2 mM, 0.1-1 mM, 0.1-0.5
mM, 0.1-0.4 mM, 0.1-0.3 mM or 0.1-0.2 mM; or at a total
concentration of 0.2-5 mM, such as 0.2-4 mM, 0.2-3 mM, 0.2-2 mM,
0.2-1 mM, 0.2-0.5 mM, 0.2-0.4 mM or 0.2-0.3 mM; or at a total
concentration of 0.3-5 mM, such as 0.3-4 mM, 0.3-3 mM, 0.3-2 mM,
0.3-1 mM, 0.3-0.5 mM or 0.3-0.4 mM; or at a total concentration of
0.4-5 mM, such as 0.4-4 mM, 0.4-3 mM, 0.4-2 mM, 0.4-1 mM or 0.4-0.5
mM; or at a total concentration of 0.5-5 mM, such as 0.5-4 mM,
0.5-3 mM, 0.5-2 mM or 0.5-1 mM.
[0139] In one aspect of the invention is provided an aqueous
solution composition of pH 5.0-8.0, suitably pH 6.0-7.0, comprising
or consisting of: [0140] daptomycin or an analogue thereof, or a
salt thereof, at a concentration of 30-60 mg/ml, such as 35-60
mg/ml, 40-60 mg/ml or 45-55 mg/ml; [0141] a divalent metal cation
selected from calcium and magnesium; [0142] a tonicity modifier;
and [0143] optionally one or more buffers being substances having
at least one ionisable group with a pK.sub.a in the range 3.0 to
9.0, suitably with pK.sub.a in the range 4.0 to 9.0; and which
pK.sub.a is within 2 pH units of the pH of the composition;
[0144] wherein the buffers are present at a total concentration of
0-5 mM, such as 0-4 mM, 0-3 mM, 0-2 mM, 0-1 mM, 0-0.5 mM, 0-0.4 mM,
0-0.3 mM, 0-0.2 mM or 0-0.1 mM; or at a total concentration of
0.1-5 mM, such as 0.1-4 mM, 0.1-3 mM, 0.1-2 mM, 0.1-1 mM, 0.1-0.5
mM, 0.1-0.4 mM, 0.1-0.3 mM or 0.1-0.2 mM; or at a total
concentration of 0.2-5 mM, such as 0.2-4 mM, 0.2-3 mM, 0.2-2 mM,
0.2-1 mM, 0.2-0.5 mM, 0.2-0.4 mM or 0.2-0.3 mM; or at a total
concentration of 0.3-5 mM, such as 0.3-4 mM, 0.3-3 mM, 0.3-2 mM,
0.3-1 mM, 0.3-0.5 mM or 0.3-0.4 mM; or at a total concentration of
0.4-5 mM, such as 0.4-4 mM, 0.4-3 mM, 0.4-2 mM, 0.4-1 mM or 0.4-0.5
mM; or at a total concentration of 0.5-5 mM, such as 0.5-4 mM,
0.5-3 mM, 0.5-2 mM or 0.5-1 mM.
[0145] In one aspect of the invention is provided an aqueous
solution composition of pH 5.0-8.0, suitably pH 6.0-7.0, comprising
or consisting of: [0146] daptomycin or an analogue thereof, or a
salt thereof, at a concentration of 30-60 mg/ml, such as 35-60
mg/ml, 40-60 mg/ml or 45-55 mg/ml; [0147] a divalent metal cation
selected from calcium and magnesium; [0148] a charged tonicity
modifier, such as sodium chloride, at a concentration of 300 mM or
more e.g. 500 mM or more e.g. 500-3000 mM e.g. 1000-2500 mM e.g.
e.g. 1500-2500 mM e.g. around 2000 mM; and [0149] optionally one or
more buffers being substances having at least one ionisable group
with a pK.sub.a in the range 3.0 to 9.0, suitably with pK.sub.a in
the range 4.0 to 9.0; and which pK.sub.a is within 2 pH units of
the pH of the composition;
[0150] wherein the buffers are present at a total concentration of
0-5 mM, such as 0-4 mM, 0-3 mM, 0-2 mM, 0-1 mM, 0-0.5 mM, 0-0.4 mM,
0-0.3 mM, 0-0.2 mM or 0-0.1 mM; or at a total concentration of
0.1-5 mM, such as 0.1-4 mM, 0.1-3 mM, 0.1-2 mM, 0.1-1 mM, 0.1-0.5
mM, 0.1-0.4 mM, 0.1-0.3 mM or 0.1-0.2 mM; or at a total
concentration of 0.2-5 mM, such as 0.2-4 mM, 0.2-3 mM, 0.2-2 mM,
0.2-1 mM, 0.2-0.5 mM, 0.2-0.4 mM or 0.2-0.3 mM; or at a total
concentration of 0.3-5 mM, such as 0.3-4 mM, 0.3-3 mM, 0.3-2 mM,
0.3-1 mM, 0.3-0.5 mM or 0.3-0.4 mM; or at a total concentration of
0.4-5 mM, such as 0.4-4 mM, 0.4-3 mM, 0.4-2 mM, 0.4-1 mM or 0.4-0.5
mM; or at a total concentration of 0.5-5 mM, such as 0.5-4 mM,
0.5-3 mM, 0.5-2 mM or 0.5-1 mM.
[0151] In one aspect of the invention is provided an aqueous
solution composition of pH 5.0-8.0, suitably pH 6.0-7.0, comprising
or consisting of: [0152] daptomycin or an analogue thereof, or a
salt thereof, at a concentration of 30-60 mg/ml, such as 35-60
mg/ml, 40-60 mg/ml or 45-55 mg/ml; [0153] a divalent metal cation
selected from calcium and magnesium; [0154] an amino acid, suitably
selected from the group consisting of consisting of glycine,
proline, methionine, arginine, lysine and aspartic acid; and [0155]
optionally one or more buffers being substances having at least one
ionisable group with a pK.sub.a in the range 3.0 to 9.0, suitably
with pK.sub.a in the range 4.0 to 9.0; and which pK.sub.a is within
2 pH units of the pH of the composition;
[0156] wherein the buffers are present at a total concentration of
0-5 mM, such as 0-4 mM, 0-3 mM, 0-2 mM, 0-1 mM, 0-0.5 mM, 0-0.4 mM,
0-0.3 mM, 0-0.2 mM or 0-0.1 mM; or at a total concentration of
0.1-5 mM, such as 0.1-4 mM, 0.1-3 mM, 0.1-2 mM, 0.1-1 mM, 0.1-0.5
mM, 0.1-0.4 mM, 0.1-0.3 mM or 0.1-0.2 mM; or at a total
concentration of 0.2-5 mM, such as 0.2-4 mM, 0.2-3 mM, 0.2-2 mM,
0.2-1 mM, 0.2-0.5 mM, 0.2-0.4 mM or 0.2-0.3 mM; or at a total
concentration of 0.3-5 mM, such as 0.3-4 mM, 0.3-3 mM, 0.3-2 mM,
0.3-1 mM, 0.3-0.5 mM or 0.3-0.4 mM; or at a total concentration of
0.4-5 mM, such as 0.4-4 mM, 0.4-3 mM, 0.4-2 mM, 0.4-1 mM or 0.4-0.5
mM; or at a total concentration of 0.5-5 mM, such as 0.5-4 mM,
0.5-3 mM, 0.5-2 mM or 0.5-1 mM.
[0157] The presently claimed invention derives from the surprising
observation that compositions comprising daptomycin are stabilized
by the addition of divalent metal cations. Such solutions may be
further stabilized by minimising the buffer concentration.
[0158] Suitably the composition of the invention remains as a clear
solution following storage at 2-8.degree. C. for an extended period
of time, such as at least 6 months, preferably at least 12 months,
most preferably at least 18 months.
[0159] Suitably the composition of the invention remains as a clear
solution following storage at 25.degree. C. for at least 2 weeks,
preferably at least 4 weeks, most preferably at least 8 weeks.
[0160] Suitably the composition of the invention has improved
storage stability either at 2-8.degree. C. or at increased
temperature than in an equivalent composition that comprises higher
concentration of the same buffer or buffers.
[0161] Suitably the composition of the invention has improved
storage stability either at 2-8.degree. C. or at increased
temperature than in an equivalent composition that does not
comprise divalent metal cations.
[0162] In one embodiment, the composition of the invention
comprises no more than 5% total impurities, such as no more than
4%, such as no more than 3%, such as no more than 2% total
impurities (by total weight of daptomycin in the composition, as
measured by RP-HPLC (Reversed-Phase High-Performance Liquid
Chromatography) or a similar suitable technique) following storage
at 2-8.degree. C. for at least 6 months, preferably at least 12
months, most preferably at least 18 months.
[0163] In one embodiment, the composition of the invention
comprises no more than 5% total impurities, such as no more than
4%, such as no more than 3%, such as no more than 2% total
impurities (by total weight of daptomycin in the composition, as
measured by RP-HPLC or a similar suitable technique) following
storage at 25.degree. C. for at least 2 weeks, preferably at least
4 weeks, most preferably at least 8 weeks.
[0164] In one embodiment, the composition of the invention
comprises lower level of impurities than a commercially available
composition comprising daptomycin (as measured by RP-HPLC or a
similar suitable technique) following storage at 2-8.degree. C. for
at least 6 months, preferably at least 12 months, most preferably
at least 18 months. In one embodiment, the commercially available
composition is Cubicin.RTM..
[0165] In one embodiment, the composition of the invention
comprises lower level of impurities than a commercially available
composition comprising daptomycin (as measured by RP-HPLC or a
similar suitable technique) following storage at 25.degree. C. for
at least 2 weeks, preferably at least 4 weeks, most preferably at
least 8 weeks. In one embodiment, the commercially available
composition is Cubicin.RTM..
[0166] In a further aspect of the invention, there is provided a
method of improving the stability of an aqueous solution
composition comprising daptomycin of pH in the range 5.0 to 8.0,
which comprises adding to the composition a divalent metal cation
and maintaining a low buffer concentration, wherein the buffer(s)
is(are) a substance having at least one ionisable group with a
pK.sub.a in the range 3.0 to 9.0 and which pK.sub.a is within 2 pH
units of the pH of the composition; and wherein the buffer(s)
is(are) present at a total concentration of 0-5 mM.
[0167] In a further aspect of the invention, there is provided the
use of divalent metal cations for improving the stability of an
aqueous solution composition comprising daptomycin of pH in the
range 5.0 to 8.0, wherein the composition optionally comprises one
or more buffers being substances having at least one ionisable
group with a pK.sub.a in the range 3.0 to 9.0 and which pK.sub.a is
within 2 pH units of the pH of the composition; and wherein the
buffers are present at a total concentration of 0-5 mM.
[0168] In a further aspect of the invention, there is provided the
use of a minimal amount of buffer for improving the stability of an
aqueous solution composition of daptomycin and a divalent metal
cation of pH in the range 5.0 to 8.0, wherein composition comprises
one or more buffers being substances having at least one ionisable
group with a pK.sub.a in the range 3.0 to 9.0 and which pK.sub.a is
within 2 pH units of the pH of the composition; and wherein the
buffers are present at a total concentration of 0-5 mM.
[0169] In an embodiment, the composition of the invention is a
composition for use in therapy. In an embodiment, the composition
of the invention is a pharmaceutical composition.
[0170] All embodiments described above with respect to the aqueous
solution composition apply equally to methods and uses of the
invention.
[0171] 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, a pre-filled syringe, a pre-filled infusion bag,
or a cartridge designed to be a replaceable item for use with an
injection device. In one embodiment, there is provided a vial
containing a composition as described herein, suitably a 10 mL
vial, suitably comprising 500 mg of daptomycin.
[0172] The compositions of the invention may suitably be packaged
for injection, especially intravenous infusion or intravenous
injection.
[0173] A desirable presentation of a stable liquid daptomycin
formulation is a 50 mg/ml product formulated either in a vial or in
a pre-filled syringe. The product will be ready to administer by IV
injection without a further dilution (e.g. over a 2 minute period).
Alternatively, the product will be administered by an IV infusion
(e.g. a 30 minute infusion) following dilution to a required
concentration. The daptomycin concentration in the diluted solution
may vary depending on the indication and the patient receiving the
infusion, e.g. between 0.5-10 mg/ml, e.g. 1-5 mg/ml. According to
certain embodiments, compositions of the invention are hypertonic
but may nevertheless be tolerable for direct injection without
dilution. Compositions of the invention containing a concentration
of charged tonicity modifier (e.g. sodium chloride) of about 500 mM
or more, particularly 1 M or more are suitably diluted before
administration. Example diluents include water for injection,
isotonic saline and isotonic dextrose solution.
[0174] The compositions of the invention may have a wide range of
osmolarity values, including hypotonic, isotonic and hypertonic
compositions. When intended for direct administration (e.g. IV
injection without dilution), suitably, the compositions of the
invention are substantially isotonic. In one embodiment, the
compositions of the invention are isotonic. Suitably, the
osmolarity of the compositions of the invention is selected to
minimize pain according to the route of administration e.g. upon
injection. Suitable compositions of the invention when intended for
direct administration have an osmolarity (excluding any
contribution to osmolarity from daptomycin) in the range of about
200 mOsm/L to about 500 mOsm/L. Suitably, the osmolarity (excluding
any contribution to osmolarity from daptomycin) is in the range of
about 250 mOsm/L to about 350 mOsm/L, more suitably, about 300
mOsm/L.
[0175] Compositions of the invention are useful for the treatment
of bacterial infections e.g. complicated skin and skin structure
infections (cSSSI), or Staphylococcus Aureus bloodstream infections
(bacteremia).
[0176] Compositions of the invention are expected to have good
physical and chemical stability as described herein.
Additional Clauses of the Invention
[0177] 1. An aqueous solution composition of pH in the range 5.0 to
8.0 comprising: [0178] daptomycin or an analogue thereof, or a salt
thereof; [0179] a divalent metal cation; and [0180] optionally one
or more buffers being substances having at least one ionisable
group with a pK.sub.a in the range 3.0 to 9.0 and which pK.sub.a is
within 2 pH units of the pH of the composition;
[0181] wherein the buffers are present at a total concentration of
0-5 mM.
[0182] 2. An aqueous solution composition according to clause 1,
wherein the concentration of daptomycin, or the analogue thereof,
or the salt thereof, is >25 mg/ml, such as .gtoreq.27 mg/ml,
.gtoreq.30 mg/ml, .gtoreq.35 mg/ml, .gtoreq.40 mg/ml, .gtoreq.45
mg/ml or .gtoreq.50 mg/ml.
[0183] 3. An aqueous solution composition according to clause 1,
wherein the concentration of daptomycin, or the analogue thereof,
or the salt thereof, is 30-100 mg/ml, such as 30-60 mg/ml, such as
35-60 mg/ml, 40-60 mg/ml or 45-55 mg/ml.
[0184] 4. An aqueous solution composition according to any one of
clauses 1 to 3, wherein the composition comprises water in an
amount of at least 65% (w/v), such as at least 70% (w/v), at least
75% (w/v), at least 80% (w/v), at least 85% (w/v), or at least 90%
(w/v).
[0185] 5. An aqueous solution composition according to any one of
clauses 1 to 4, wherein the concentration of divalent metal cation
is 10-150 mM, such as 20-130 mM, 30-120 mM, 40-100 mM, 40-80 mM or
50-70 mM.
[0186] 6. An aqueous solution composition according to any one of
clauses 1 to 5, wherein the divalent metal cation is selected from
calcium, magnesium, manganese and zinc ions.
[0187] 7. An aqueous solution composition according to clause 6,
wherein the divalent metal cation is calcium ion.
[0188] 8. An aqueous solution composition according to clause 6,
wherein the divalent metal cation is magnesium ion.
[0189] 9. An aqueous solution composition according to any one of
clauses 1 to 8, wherein the total concentration of buffers is 0-4
mM such as 0-3 mM, 0-2 mM, 0-1 mM, 0-0.5 mM, 0-0.4 mM, 0-0.3 mM,
0-0.2 mM or 0-0.1.
[0190] 10. An aqueous solution composition according to any one of
clauses 1 to 8, wherein the aqueous solution composition is
substantially free of buffer.
[0191] 11. An aqueous solution composition according to any one of
clauses 1 to 9, wherein the buffer or buffers is/are selected from
the group consisting of maleate, sulphite, aspartame, aspartate,
glutamate, tartrate, gluconate, adenine, succinate, ascorbate,
benzoate, phenylacetate, gallate, cytosine, p-aminobenzoic acid,
sorbate, acetate, propionate, alginate, urate,
2-(N-morpholino)ethanesulphonic acid, bicarbonate,
bis(2-hydroxyethyl) iminotris(hydroxymethyl)methane,
N-(2-acetamido)-2-iminodiacetic acid,
2-[(2-amino-2-oxoethyl)amino]ethanesulphonic acid,
piperazine-N,N'-bis(2-ethanesulphonic acid) (PIPES), phosphate,
N,N-bis(2-hydroxyethyl)-2-aminoethanesulphonic acid,
3-[N,N-bis(2-hydroxyethyl)amino]-2-hydroxypropanesulphonic acid,
triethanolamine, piperazine-N,N'-bis(2-hydroxypropanesulphonic
acid), tris(hydroxymethyl)aminomethane,
N-tris(hydroxymethyl)glycine,
N-tris(hydroxymethyl)methyl-3-aminopropanesulphonic acid, citrate
and salts thereof, and combinations thereof.
[0192] 12. An aqueous solution composition according to clause 11,
wherein the buffer is selected from the group consisting of
maleate, tartrate, lactate, benzoate, acetate, bicarbonate,
phosphate, tris(hydroxymethyl)aminomethane and citrate, especially
phosphate.
[0193] 13. An aqueous solution composition according to any one of
clauses 1 to 12, wherein the pH is in the range 6.0 to 7.0, and the
composition optionally comprises one or more buffers being
substances having at least one ionisable group with a pK.sub.a in
the range 4.0 to 9.0 and which pK.sub.a is within 2 pH units of the
pH of the composition.
[0194] 14. An aqueous solution composition according to any one of
clauses 1 to 13, further comprising a stabilizer in a stabilizing
amount.
[0195] 15. An aqueous solution composition according to any one of
clauses 1 to 14, further comprising a surfactant.
[0196] 16. An aqueous solution composition according to clause 15,
wherein the surfactant is present at a concentration of 0.01-10
mg/ml, such as 0.05-2 mg/ml.
[0197] 17. An aqueous solution composition according to clause 15
or clause 16, wherein the surfactant is a non-ionic surfactant.
[0198] 18. An aqueous solution composition according to clause 17,
wherein the non-ionic surfactant is selected from the group
consisting of an alkyl glycoside, a polysorbate, an alkyl ether of
polyethylene glycol, a block copolymer of polyethylene glycol and
polypropylene glycol, and an alkylphenyl ether of polyethylene
glycol.
[0199] 19. An aqueous solution composition according to clause 17
or clause 18, wherein the non-ionic surfactant is present at a
concentration of 10-10,000 .mu.g/ml, such as 10-8,000 .mu.g/ml,
10-5,000 .mu.g/ml, 10-3,000 .mu.g/ml, 10-2000 .mu.g/ml, 50-1000
.mu.g/ml, 100-500 .mu.g/ml or about 200 .mu.g/ml.
[0200] 20. An aqueous solution composition according to clause 15
or clause 16, wherein the surfactant is a cationic surfactant.
[0201] 21. An aqueous solution composition according to clause 20,
wherein the cationic surfactant is selected from benzalkonium and
benzethonium salts.
[0202] 22. An aqueous solution composition according to clause 15
or clause 16, wherein the surfactant is an anionic surfactant such
as sodium dodecyl sulfate, ammonium dodecyl sulfate, a carboxylate
salt (sodium or ammonium salt of stearic or palitic acid), or an
ether phosphate.
[0203] 23. An aqueous solution composition according to clause 15
or clause 16, wherein the surfactant is a zwitterionic surfactant
such as phosphatidylcholine, phosphatidylserine,
phosphatitylethanolamine or
3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate
(CHAPS).
[0204] 24. An aqueous solution composition according to any one of
clauses 1 to 23, further comprising an antioxidant.
[0205] 25. An aqueous solution composition according to clause 24,
wherein the antioxidant is selected from the group consisting of
methionine, glutathione, ascorbate, butylated hydroquinone,
lactate, nicotinamide, nicotinate, tryptophan, phenylalanine and
tyrosine; and in particular is methionine.
[0206] 26. An aqueous solution composition according to clause 24
or clause 25, wherein the antioxidant is present at a concentration
of 1-100 mg/ml, such as 5-50 mg/ml.
[0207] 27. An aqueous solution composition according to any one of
clause 1 to 26, further comprising a chelating agent.
[0208] 28. An aqueous solution composition according to clause 27,
wherein the chelating agent is selected from the group consisting
of EDTA, EGTA and citrate.
[0209] 29. An aqueous solution composition according to clause 27
or clause 28, wherein the chelating agent is present at a
concentration of 0.1-20 mM, such as 0.1-15 mM, 0.1-10 mM, 0.1-5 mM
or 0.1-1 mM.
[0210] 30. An aqueous solution composition according to any one of
clauses 1 to 29, further comprising a polyanion.
[0211] 31. An aqueous solution composition according to clause 30,
wherein the polyanion is selected from the group consisting of
carboxymethyl cellulose and dextran sulfate.
[0212] 32. An aqueous solution composition according to clause 31,
wherein the polyanion is present at a concentration of 1-10 mg/ml,
such as 1-5 mg/ml.
[0213] 33. An aqueous solution composition according to any one of
clauses 1 to 32, further comprising a polycation.
[0214] 34. An aqueous solution composition according to clause 33,
wherein the polycation is an oligomer of ethyleneimine selected
from the group consisting of diethylenetriamine,
triethylenetetramine (TETA), tetraethylenepentamine and
pentaethylenehexamine; and in particular is TETA.
[0215] 35. An aqueous solution composition according to clause 33
or clause 34, wherein the polycation is present at a concentration
of 1-10 mg/ml, such as 1-5 mg/ml.
[0216] 36. An aqueous solution composition according to any one of
clauses 1 to 35, further comprising a charged amphiphilic
species.
[0217] 37. An aqueous solution composition according to clause 36,
wherein the charged amphiphilic species is selected from the group
consisting of benzoic acid, 2-hydroxybenzoic acid, 3-hydroxybenzoic
acid, 4-hydroxybenzoic acid, 3,4,5-trihydroxybenzoic acid,
4-hydroxy-3-methoxybenzoic acid, phthalic acid, isophthalic acid,
terephthalic acid, 1-naphthoic acid, 2-naphthoic acid,
indole-3-acetic acid, phenylacetic acid, 3-phenylpropionic acid,
trans-cinnamic acid, cis-cinnamic acid and mandelic acid, and in
particular is benzoic acid.
[0218] 38. An aqueous solution composition according to clause 36
or clause 37, wherein the charged amphiphilic species is present at
a concentration of 1-100 mM, such as 5-20 mM.
[0219] 39. An aqueous solution composition according to any one of
clauses 1 to 38, further comprising a tonicity modifier.
[0220] 40. An aqueous solution composition according to clause 39,
wherein the tonicity modifier is an uncharged tonicity modifier and
is selected from the group consisting of sugars, sugar alcohols,
other polyols and polyethylene glycols.
[0221] 41. An aqueous solution composition according to clause 40,
wherein the uncharged tonicity modifier is selected from the group
consisting of glycerol, 1,2-propanediol, mannitol, sorbitol,
sucrose, trehalose, lactose, PEG300 and PEG400.
[0222] 42. An aqueous solution composition according to clause 40
or clause 41, wherein the concentration of the uncharged tonicity
modifier is 50-5000 mM, such as 100-1000 mM, such as 100-500 mM,
such as 200-400 mM, such as about 300 mM.
[0223] 43. An aqueous solution composition according to clause 39,
wherein the tonicity modifier is a charged tonicity modifier and is
suitably selected from sodium chloride, sodium sulphate, and an
amino acid e.g. glycine or arginine especially sodium chloride.
[0224] 44. An aqueous solution composition according to clause 43,
wherein the concentration of the charged tonicity modifier is
50-2000 mM, 50-1000 mM, such as 50-500 mM such as 100-200 mM, such
as about 150 mM.
[0225] 45. An aqueous solution composition according to clause 43,
wherein the concentration of the charged tonicity modifier is 300
mM or more e.g. 500 mM or more e.g. 500-3000 mM e.g. 1000-2500 mM
e.g. e.g. 1500-2500 mM e.g. around 2000 mM 46. An aqueous solution
composition according to any one of clauses 1 to 45, further
comprising an amino acid.
[0226] 47. An aqueous solution composition according to clause 46,
wherein the amino acid is selected from the group consisting of
glycine, proline, methionine, arginine, lysine, aspartic acid and
glutamic acid, in particular selected from the group consisting of
glycine, proline, methionine, arginine, lysine and aspartic
acid.
[0227] 48. An aqueous solution composition according to clause 46
or clause 47, wherein the amino acid is present at a concentration
of 1-200 mM, such as 1-100 mM, 1-50 mM, 1-20 mM, 1-10 mM, 1-5 mM,
1-4 mM, 1-3 mM or 1-2 mM.
[0228] 49. An aqueous solution composition according to any one of
clauses 1 to 48, wherein composition comprises no more than 5%
total impurities, such as no more than 4% total impurities, no more
than 3% total impurities, or no more than 2% total impurities (by
total weight of daptomycin in the composition, as measured by
RP-HPLC) following storage at 2-8.degree. C. for at least 6 months,
preferably at least 12 months, most preferably at least 18
months.
[0229] 50. An aqueous solution composition according to any one of
clauses 1 to 49, wherein the composition comprises no more than 5%
total impurities, such as no more than 4% total impurities, no more
than 3% total impurities, or no more than 2% total impurities (by
total weight of daptomycin in the composition, as measured by
RP-HPLC) following storage at 25.degree. C. for at least 2 weeks,
preferably at least 4 weeks, most preferably at least 8 weeks.
[0230] 51. An aqueous solution composition of pH in the range 5.0
to 8.0 comprising: [0231] daptomycin or an analogue thereof, or a
salt thereof; [0232] a divalent metal cation; and [0233] optionally
one or more buffers being substances having at least one ionisable
group with a pK.sub.a in the range 3.0 to 9.0 and which pK.sub.a is
within 2 pH units of the pH of the composition;
[0234] wherein the buffers are present at a total concentration of
0-5 mM; wherein the concentration of daptomycin, or the analogue
thereof, or the salt thereof, in the composition is 30 mg/ml and
wherein the composition comprises water in an amount of at least
90% (w/v).
[0235] 52. An aqueous solution composition according to clause 51
wherein the pH is in the range 6.0 to 7.0.
[0236] 53. An aqueous solution composition according to clause 51
or clause 52 wherein the aqueous solution composition is
substantially free of buffer.
[0237] 54. An aqueous solution composition according to clause 51
or clause 52 wherein the total concentration of buffers is 0.5-5
mM, such as 0.5-4 mM, 0.5-3 mM, 0.5-2 mM or 0.5-1 mM.
[0238] 55. An aqueous solution composition according to any one of
clauses 51, 52 and 54 wherein the buffer is phosphate.
[0239] 56. An aqueous solution composition according to any one of
clauses 51 to 55 comprising a charged tonicity modifier such as
sodium chloride at a concentration of 300 mM or more e,g, 500 mM or
more
[0240] 57. An aqueous solution composition according to any one of
clauses 1 to 56, which is a composition for use in therapy.
[0241] 58. An aqueous solution composition according to any one of
clauses 1 to 57, which is a pharmaceutical composition.
[0242] 59. An aqueous solution composition according to any one of
clauses 1 to 58 for use in the treatment of bacterial
infections.
[0243] 60. A method of treatment of bacterial infections in a
patient, such as a human patient, which comprises administering to
a patient in need thereof an effective amount of an aqueous
solution composition according to any one of clauses 1 to 58.
[0244] 61. Use of an aqueous solution composition according to any
one of clauses 1 to 58 for the manufacture of a medicament for the
treatment of bacterial infections.
EXAMPLES
[0245] General Methods
[0246] Reversed-Phase Chromatography (RP-HPLC)--Method 1
[0247] Ultra-high performance reverse phase chromatography was
performed using the Waters ACQUITY H-class Bio UPLC.RTM. system
with a 5 .mu.m Zorba Eclipse XDB-C18 150.times.4.6 mm column.
Mobile Phase A (50 mM sodium phosphate in MQW) and Mobile Phase B
(50% CAN, 25 mM sodium phosphate in MQW) were used in gradient
elution. Injection volume was 10 .mu.l and flow rate was 1 mL/min,
with 214 nm UV detection. All analyses were performed at 60.degree.
C.
[0248] Reversed-Phase Chromatography (RP-HPLC)--Method 2
[0249] Ultra-high performance reverse phase chromatography was
performed using the Thermo-scientific Ultimate 3000 UPLC.RTM.
system with a 5 .mu.m Kinetix C8 250.times.4.6 mm column. Isocratic
elution was used. The mobile phase consisted of 50 mM ammonium
dihydrogen phosphate in MQW, pH 5.0 (70%) and acetonitrile (30%).
Injection volume was 10 .mu.l and flow rate was 1.5 mL/min, with
214 nm UV detection. All analyses were performed at 25.degree.
C.
Example 1--Effect of Calcium Cations and Other Formulation
Conditions on the Stability of Daptomycin (50 mg/ml)
[0250] Stability of daptomycin (50 mg/ml) was assessed using the
RP-HPLC method described in General Methods, following incubation
at 25.degree. C. (8 weeks) and 2-8.degree. C. for 26 weeks in the
compositions shown in Table 1 and 2.
TABLE-US-00001 TABLE 1 Stability of daptomycin (50 mg/ml) following
incubation at 2-8.degree. C. for 26 weeks. RP- HPLC method 1 was
used and stability is expressed as % retention of the main peak vs.
time zero Calcium Magnesium Formulation Phosphate ADA* chloride
chloride Arginine % RP-HPLC main peak No. (mM) (mM) (mM) (mM) (mM)
pH (vs. time zero) 1-1 -- -- -- -- -- 6.0 84.62 1-2 -- -- 30 -- --
6.0 94.98 1-3 -- -- 60 -- -- 6.0 98.98 1-4 -- -- 120 -- -- 6.0
99.91 1-5 -- -- -- 30 -- 6.0 94.08 1-6 -- -- -- 60 -- 6.0 95.74 1-7
-- -- -- 120 -- 6.0 98.01 1-8 -- 50 60 -- -- 6.0 96.26 1-9 50 -- 60
-- -- 6.0 96.00 1-10 -- -- 60 -- 50 6.0 97.03 1-11 -- -- 60 -- --
7.0 98.86 1-12 -- -- 60 -- -- 8.0 95.11 1-13 -- -- 60 -- -- 5.0
93.12 1-14 -- -- 60 -- -- 4.0 81.35 *ADA =
2-[(2-amino-2-oxoethyl)-(carboxymethyl)amino]acetic acid.
TABLE-US-00002 TABLE 2 Stability of daptomycin (50 mg/ml) following
incubation at 25.degree. C. for 8 weeks. RP-HPLC method 1 was used
and stability is expressed as % retention of the main peak vs. time
zero. Calcium Magnesium % RP-HPLC main peak Formulation chloride
chloride (vs. time zero) No. (mM) (mM) pH 25.degree. C. (8 weeks)
1-1 -- -- 6.0 71.7 1-2 30 -- 6.0 84.2 1-3 60 -- 6.0 90.0 1-4 120 --
6.0 90.4 1-5 -- 30 6.0 79.6 1-6 -- 60 6.0 84.0 1-7 -- 120 6.0
87.7
[0251] It was shown that the presence of calcium chloride improved
the stability of daptomycin at pH 6.0. The magnitude of the
improvement increased with increasing concentration of calcium in
the composition. The presence of magnesium chloride also appeared
to improve the stability of daptomycin in a concentration-dependent
manner, although the magnitude of the improvement was lower than
that brought about by calcium at the same concentration.
[0252] Interestingly, it was also demonstrated that the presence of
a 50 mM buffer (phosphate or ADA) resulted in an impairment in
stability in the presence of 60 mM calcium. This is in contrast
with the results reported in WO2011/035108A1. Similarly, the
presence of arginine at 50 mM also led to an impairment of
daptomycin stability, which is in contrast with the results
reported in WO2011/062676A1.
[0253] Lastly, it was shown that the optimal pH for daptomycin (50
mg/ml) stability in the presence of calcium is approximately
6.0-7.0.
Example 2--Effect of pH and Additional Formulation Parameters on
the Stability of Daptomycin (50 mg/ml) in the Presence of Calcium
Chloride
[0254] Stability of daptomycin (50 mg/ml) was assessed using the
RP-HPLC method 2 described in General Methods, following incubation
at 25.degree. C. (8 weeks) in the compositions shown in Table
3.
TABLE-US-00003 TABLE 3 Stability of daptomycin (50 mg/ml) following
incubation at 25.degree. C. for 8 weeks. RP-HPLC method 2 was used
and stability is expressed as % retention of the main peak vs. time
zero. Calcium Sodium % RP-HPLC main peak Formulation chloride
chloride Glycerol 1,2-propanediol (vs. time zero) No. (mM) (mM)
(mM) (mM) pH 25.degree. C. (8 weeks) 2-1 60 -- -- -- 5.0 65.2 2-2
60 -- -- -- 6.0 89.4 2-3 60 -- -- -- 7.0 93.1 2-4 60 -- -- -- 8.0
82.3 2-5 60 150 -- -- 6.0 89.9 2-6 60 2000 -- -- 6.0 95.9 2-7 60 --
300 -- 6.0 89.1 2-8 60 -- -- 300 6.0 89.2 2-9 60 -- -- 2000 6.0
89.8
[0255] In this experiment it was shown that the pH optimum for
daptomycin stability in the presence of calcium chloride is around
7.0. The stability at pH 7.0 was better than at pH 6.0 and
considerably better than at pH 5.0 and 8.0. Addition of a polyol
(glycerol at 300 mM or 1,2-propanediol at 300 mM and 2000 mM) had a
limited impact on daptomycin stability in the presence of calcium
chloride at pH 6.0. Addition of 150 mM sodium chloride also had a
limited impact on daptomycin stability in the presence of calcium
chloride at pH 6.0. In contrast, addition of a higher concentration
(2000 mM) of sodium chloride resulted in further considerable
improvement in stability of daptomycin stability in the presence of
calcium chloride.
Example 3--Effect of pH and Additional Formulation Parameters on
the Stability of Daptomycin (50 mg/ml) in the Presence of Calcium
Chloride
[0256] Stability of daptomycin (50 mg/ml) was assessed using the
RP-HPLC method 2 described in General Methods, following incubation
at 25.degree. C. (8 weeks) in the compositions shown in Table
4.
TABLE-US-00004 TABLE 4 Stability of daptomycin (50 mg/ml) following
incubation at 25.degree. C. for 8 weeks. RP-HPLC method 2 was used
and stability is expressed as % retention of the main peak vs. time
zero. Calcium Sodium % RP-HPLC main peak Formulation chloride
chloride (vs. time zero) No. (mM) (mM) pH 25.degree. C. (8 weeks)
3-1 60 -- 6.0 92.6 3-2 60 -- 6.5 91.2 3-3 60 -- 7.0 88.9 3-4 60 150
6.0 92.9 3-5 60 500 6.0 93.9 3-6 60 1000 6.0 94.8 3-7 60 2000 6.0
94.9 3-8 60 150 7.0 88.8 3-9 60 500 7.0 89.6 3-10 60 1000 7.0 92.1
3-11 60 2000 7.0 93.8
[0257] In this experiment it was shown that the pH optimum for
daptomycin stability in the presence of calcium chloride is around
6.0. The stability at pH 6.0 was better than at pH 6.5 and at 7.0.
The results show that the stability of the formulation
progressively increased as the concentration of sodium chloride was
increased from 150 mM to 2000 mM.
Example 4--Effect of Buffers on the Stability of Daptomycin (50
mg/ml) in the Presence of Calcium Chloride
[0258] Study A
[0259] Stability of daptomycin (50 mg/ml) was assessed using the
RP-HPLC method 2 described in General Methods, following incubation
at 25.degree. C. (8 weeks) in the compositions shown in Table
5.
TABLE-US-00005 TABLE 5 Stability of daptomycin (50 mg/ml) following
incubation at 25.degree. C. for 8 weeks. RP-HPLC method 2 was used
and stability is expressed as % retention of the main peak vs. time
zero. Calcium Sodium % RP-HPLC main peak Formulation chloride
Histidine phosphate ADA PIPES (vs. time zero) No. (mM) (mM) (mM)
(mM) (mM) pH 25.degree. C. (8 weeks) 4-1 60 -- -- -- 6.0 89.4 4-2
60 20 -- -- 6.0 90.4 4-3 60 -- 20 -- 6.0 89.1 4-4 60 -- -- 20 6.0
88.9 4-5 60 -- -- -- 20 6.0 89.9
[0260] In this study, the presence of a buffer at a concentration
of 20 mM did not appear to impact on chemical stability to a very
significant effect. The formulation without buffer was very stable.
It was noted that the composition containing ADA at a concentration
of 20 mM contained visible particles after storage at 25.degree. C.
for 8 weeks whereas the other formulations presented in Table 5 did
not show any visible particles after corresponding storage.
Therefore, ADA appears to impair physical stability in this
composition.
[0261] Study B
[0262] A more thorough study was undertaken to investigate the
effect of histidine in formulations of daptomycin. The source of
daptomycin was different from that of Study A. Stability of
daptomycin (50 mg/ml) was assessed using the RP-HPLC method 2
described in General Methods, following incubation at 25.degree. C.
(8 weeks) in the compositions shown in Table 6.
TABLE-US-00006 TABLE 6 Stability of daptomycin (50 mg/ml) following
incubation at 25.degree. C. for 8 weeks. RP-HPLC method 2 was used
and stability is expressed as % retention of the main peak vs. time
zero. Calcium % RP-HPLC main peak Formulation chloride Histidine
(vs. time zero) No. (mM) (mM) pH 25.degree. C. (8 weeks) 4-6 60 --
6.0 92.6 4-7 60 10 6.0 91.0 4-8 60 20 6.0 88.0 4-9 60 60 6.0 84.7
4-10 60 -- 7.0 88.9 4-11 60 10 7.0 86.7 4-12 60 20 7.0 84.3 4-13 60
60 7.0 82.1
[0263] This study shows that the presence of histidine buffer is
deleterious to the stability of the formulation when used at a
concentration of 10 mM or higher.
[0264] Taking the results of Study A and Study B together: Buffer
free formulations are very stable. Histidine and ADA are generally
not tolerated in the formulation and their presence appears to
reduce stability. Sodium phosphate and PIPES appear to be tolerated
at a concentration of up to 20 mM.
[0265] Throughout the specification and the claims which follow,
unless the context requires otherwise, the word `comprise`, and
variations such as `comprises` and `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.
[0266] All patents, patent applications and references mentioned
throughout the specification of the present invention are herein
incorporated in their entirety by reference.
[0267] The invention embraces all combinations of preferred and
more preferred groups and suitable and more suitable groups and
embodiments of groups recited above.
* * * * *