U.S. patent application number 15/393026 was filed with the patent office on 2017-04-20 for pharmaceutical composition.
The applicant listed for this patent is FERRING B.V.. Invention is credited to Mattias Malm, Anders Nilsson, Britta Siekmann, Kazimierz Wisniewski.
Application Number | 20170106043 15/393026 |
Document ID | / |
Family ID | 43478408 |
Filed Date | 2017-04-20 |
United States Patent
Application |
20170106043 |
Kind Code |
A1 |
Nilsson; Anders ; et
al. |
April 20, 2017 |
PHARMACEUTICAL COMPOSITION
Abstract
The present invention relates to pharmaceutical compositions
having improved stability.
Inventors: |
Nilsson; Anders; (Copenhagen
S, DK) ; Malm; Mattias; (Copenhagen S, DK) ;
Wisniewski; Kazimierz; (San Diego, CA) ; Siekmann;
Britta; (Lomma, SE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
FERRING B.V. |
Hoofddorp |
|
NL |
|
|
Family ID: |
43478408 |
Appl. No.: |
15/393026 |
Filed: |
December 28, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13824132 |
May 3, 2013 |
9566311 |
|
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PCT/IB2011/002394 |
Sep 29, 2011 |
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15393026 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 47/12 20130101;
A61P 17/02 20180101; A61P 19/10 20180101; A61P 25/22 20180101; A61K
38/12 20130101; A61K 38/095 20190101; A61P 15/14 20180101; A61P
7/04 20180101; A61P 15/00 20180101; A61P 1/04 20180101; A61P 15/04
20180101; A61P 1/00 20180101; A61P 15/10 20180101; A61K 47/26
20130101; A61P 35/00 20180101; A61P 1/14 20180101; A61K 47/02
20130101; A61P 25/04 20180101; A61P 29/00 20180101; A61K 47/183
20130101; A61K 47/20 20130101; A61K 9/0019 20130101; A61P 5/10
20180101; A61P 15/08 20180101; A61P 25/24 20180101; A61P 25/00
20180101; A61P 25/18 20180101; A61P 31/04 20180101; A61P 43/00
20180101; A61P 1/10 20180101; A61P 7/06 20180101; A61K 9/0043
20130101 |
International
Class: |
A61K 38/11 20060101
A61K038/11; A61K 47/02 20060101 A61K047/02; A61K 47/12 20060101
A61K047/12; A61K 47/20 20060101 A61K047/20; A61K 47/18 20060101
A61K047/18 |
Foreign Application Data
Date |
Code |
Application Number |
Sep 30, 2010 |
EP |
10251690.3 |
Claims
1-37. (canceled)
38. A liquid composition, comprising: a pharmaceutically active
compound selected from: a compound having the formula (I):
##STR00006## wherein: n is selected from 0, 1 and 2; p is selected
from 0, 1, 2, 3, 4, 5 and 6; R.sub.1 is selected from aryl
optionally substituted with at least one OH, F, Cl, Br, alkyl or
O-alkyl substituent; R.sub.2 is selected from R.sub.4, H, alkyl,
cycloalkyl, aryl and 5- and 6-membered heteroaromatic ring systems;
R.sub.3 is selected from H and a covalent bond to R.sub.2, when
R.sub.2 is R.sub.4, to form a ring structure; R.sub.4 is C.sub.1-6
alkylene moiety substituted with at least one O-alkyl, S-alkyl or
OH substituent; W and X are each independently selected from
CH.sub.2 and S, but may not both be CH.sub.2; alkyl is selected
from C.sub.1-6 straight and C.sub.4-8 branched chain alkyl and
optionally has at least one hydroxyl substituent; aryl is selected
from phenyl and mono- or poly-substituted phenyl; with the proviso
that when R.sub.2 is H, p is 1, R.sub.3 is H, n is 1 and W and X
are both S, R.sub.1 is not 4-hydroxyphenyl, or a solvate or
pharmaceutically acceptable salt thereof; and a compound having
structure of formula (II): ##STR00007## or a pharmaceutically
acceptable salt thereof; and an anti-oxidant; wherein the pH of the
composition is from 5.0 to 6.0.
39. The composition of claim 38, wherein the pH of the composition
is from 5.1 to 6.
40. The composition of claim 38, wherein the pH of the composition
is from 5.2 to 5.65.
41. The composition of claim 38, wherein the pH of the composition
is from 5.26 to 5.8.
42. The composition of claim 38, further comprising a buffering
agent.
43. The composition of claim 38, wherein the anti-oxidant is
methionine, EDTA, or a combination of methionine and EDTA.
44. The composition of claim 38, further comprising an isotonicity
agent.
45. The composition of claim 38, wherein the pharmaceutically
active compound is the compound of formula (II): ##STR00008## or a
pharmaceutically acceptable salt thereof.
46. The composition of claim 45, wherein the pH of the composition
is from 5.0 to 5.9.
47. The composition of claim 45, wherein the pH of the composition
is from 5.1 to 5.9.
48. The composition of claim 45, wherein the pH of the composition
is from 5.2 to 5.8.
49. The composition of claim 45, further comprising a buffer.
50. The composition of claim 49, wherein the buffer is a
citrate/phosphate buffer.
51. The composition of claim 45, wherein the concentration of the
compound of formula (II) in the liquid composition is from 0.01 to
4 mg/mL.
52. The composition of claim 45, wherein the concentration of the
compound of formula (II) in the liquid composition is from 0.05 to
2 mg/mL.
53. The composition of claim 45, wherein the concentration of the
compound of formula (II) in the liquid composition is from 0.1 to
1.4 mg/mL.
54. The composition of claim 45, wherein the concentration of the
compound of formula (II) in the liquid composition is from 0.2 to
0.7 mg/mL.
55. The composition of claim 45, further comprising an
anti-oxidant.
56. The composition of claim 45, further comprising an isotonicity
agent.
57. The composition of claim 56, wherein the isotonicity agent is
NaCl.
58. A method of treatment of compromised lactation conditions,
labour induction impairment, uterine atony conditions, excessive
bleeding, inflammation, pain, abdominal pain, back pain, male and
female sexual dysfunction, irritable bowel syndrome (IBS),
constipation, gastrointestinal obstruction, autism, stress,
anxiety, depression, anxiety disorder, surgical blood loss,
post-partum haemorrhage, wound healing, infection, mastitis,
placenta delivery impairment, or osteoporosis, or a method for the
diagnosis of cancer and placental insufficiency, comprising
administration to a patient in need thereof the composition of
claim 38.
59. The method of claim 58, wherein the method is for the treatment
of uterine atony conditions or excessive bleeding.
60. The method of claim 59, wherein the method is for the treatment
of uterine atony conditions following vaginal delivery of the
infant or delivery of the infant by Caesarean section, or in a
patient who is at risk of developing post-partum haemorrhage.
61. The method of claim 59, wherein the method is for the treatment
of excessive bleeding following vaginal delivery.
62. A kit, comprising: a liquid pharmaceutical composition of claim
38; and a container for the composition.
63. The kit of claim 62, wherein the pharmaceutically active
compound is the compound of formula (II).
Description
[0001] The present invention relates to pharmaceutical
compositions, for example pharmaceutical compositions for the
treatment of post-partum haemorrhage (PPH) or other medical
applications. In particular, it relates to pharmaceutical
compositions having improved stability, for example at room or
ambient temperature.
[0002] Postpartum haemorrhage (PPH) is one of the leading causes of
pregnancy-related mortality and severe morbidity in developing
countries as well as in the industrialised world. It is a
potentially life-threatening condition which is demonstrated by
about 140,000 deaths annually, one every four minutes, the vast
majority among women who do not have access to adequate obstetric
healthcare. Although the problem is numerically important, not all
regions of the industrialised world are similarly affected and in
Europe, the rates of death associated with maternal haemorrhage
vary widely from one country to another, A population based survey
in 11 regions within European countries showed that rates of severe
haemorrhage ranged from 0.1% to 0.9% of pregnancies (MOMS-B group,
1999). There are good reasons to believe that differences in
clinical practice may be of great importance to the differences in
morbidity/mortality. In the United Kingdom, the Confidential
Enquiry into Maternal Deaths, covering the years 1994-1996, showed
that after action had been taken in all maternity units to
establish guidelines for management of third stage and postpartum
haemorrhage, there were no deaths from haemorrhage in uncomplicated
vaginal births (Department of Health et al, 1998). This achievement
supports the assumption that clinical management has a key role in
prevention of severe maternal haemorrhage.
[0003] PPH is difficult to manage because evaluation of blood loss
in the delivery unit is unreliable. Action is often taken in
response to the development of maternal signs such as hypotension
or malaise rather than on the basis of estimated blood loss. The
delayed action is responsible for many cases of severe haemorrhage,
and immediate surgery may be required because time spent using
other treatment methods would be dangerous for the patient. These
considerations speak in favour of the application of a policy of
routine prophylactic administration of a uterotonic to all
parturients. The case for such a policy is augmented by the fact
that uterine atony is the most significant contributor to PPH.
Uterine atony is a loss of tone in the uterine musculature.
Normally, contraction of the uterine muscle compresses the vessels
and reduces flow. This increases the likelihood of coagulation and
prevents bleeds. Thus, lack of uterine muscle contraction can cause
an acute haemorrhage. Clinically, 75-80% of postpartum hemorrhages
are due to uterine atony.
[0004] Reviews have shown compelling evidence in support of routine
prophylactic administration of a uterotonic, either in isolation or
as part of the entity Active Management of the Third Stage of
Labour (AMTSL); AMTSL is usually defined as an intervention with
three components: a prophylactic administration of a uterotonic
agent, early cord clamping and controlled cord traction. Further,
AMTSL is shown to be equally effective in "low-risk" and
"high-risk" women. Today, the use of a uterotonic drug as
prophylaxis in all vaginal hospital deliveries to prevent severe
haemorrhage is routine clinical management in most of Europe, and
the practice is increasing globally.
[0005] The uterotonic drugs currently available are oxytocin,
ergometrine, Syntometrine.RTM. (a combination of oxytocin and
ergometrine) and misoprostol. However, these are not without
disadvantages. Misoprostol is administered orally or vaginally and
is less efficacious than injectable uterotonics; it is generally
recommended that it is not used where injectable uterotonics are
available. Syntometrine.RTM. is licensed in only a few countries in
Europe. It is possibly more effective than oxytocin alone but is
associated with more side effects, particularly nausea and
vomiting. Further, it is unsuitable for use in women with
hypertension, preeclampsia and heart disease, thereby reducing its
suitability for routine prophylactic usage. Oxytocin itself has the
disadvantage of a short half-life. Although this can be
circumvented by administration as a continuous intravenous infusion
to provide sustained uterotonic activity, this is more inconvenient
than a single injection. If administered as a single bolus dose,
whether as intravenous or intramuscular injection, close monitoring
of uterine tone is required and additional uterotonic medication
may be required due to the short half-life.
[0006] Carbetocin
[(1-desamino-1-monocarba-2(O-methyl)-tyrosine)oxytocin] is a
long-acting synthetic analogue of oxytocin, with agonist action.
Carbetocin (PABAL.RTM., DURATOCIN.RTM.) is currently approved for
the prevention of uterine atony following delivery of the infant by
Caesarean section under epidural or spinal anaesthesia. In its
present marketed usage, intravenous administration of carbetocin
provides a half-life of approximately 40 minutes, which is 4 to 10
times longer than the reported half-life of oxytocin (4 to 10
minutes). However, using intramuscular injection, carbetocin
reaches peak plasma concentrations in less than thirty minutes and
has a bioavailability of 80% (W Rath, European Journal of
Obstetrics and Gynaecology and Reproductive Biology 147 (2009)
15-20). Thus, carbetocin has the potential to be a near ideal drug
for routine postpartum haemorrhage prophylaxis, offered in all
hospital vaginal deliveries, because it is suitable for both
intramuscular injection and intravenous administration, offering
convenience and simple implementation; it has quick onset of
action; is long-acting, especially compared to oxytocin; is rarely
associated with adverse drug reactions; and has excellent
tolerability. In published clinical trials, carbetocin has shown
efficacy similar to oxytocin and Syntometrine.RTM. or even a trend
towards better efficacy, demonstrated on several outcomes: blood
loss (measured or estimated), incidence of blood loss >500 ml,
additional use of uterotonic medication or total uterotonic
interventions. Thus, carbetocin should offer an improvement over
currently available options for prevention of uterine atony and
excessive bleeding following vaginal delivery. In comparison with
oxytocin, its advantage is primarily that it would replace the need
for continuous infusion or additional uterotonic intervention. In
practice, carbetocin has the potential to replace 2-4 hours of
postpartum routine infusion therapy, and/or to improve outcome
versus oxytocin bolus injection. Further, any reduction in
additional interventions represents a favourable pharmacoeconomic
case for use of carcetocin. In comparison with Syntometrine.RTM.,
its advantage is primarily better tolerability and safety and lack
of important contraindications. In either case, carbetocin is
better suited for, and is likely to facilitate the implementation
of, routine prophylactic usage.
[0007] The current carbetocin formulation (PABAL.RTM. 100
micrograms/mL solution for injection, Ferring Pharmaceuticals
Limited) is not room temperature (RT) stable and requires
refrigerated storage at a temperature of 2-8.degree. C. There is,
therefore, a need for a formulation of carbetocin which is room
temperature stable (for example at 25.degree. C. and 60% relative
humidity) for up to two years, allowing, for example, use in
ambulances. This would provide an advantage in climatic zone I/II.
More importantly, there is a need for a formulation which may be
stored unrefrigerated in climatic zone III/IV (high temperature,
e.g. tropical) regions--that is meeting the temperature and
humidity stability requirements for these zones, for example, a
documented long-term temperature stability at 30.degree. C. and
relative humidity up to 75%. The climatic zone terminology is used
by the FDA and EMEA and is familiar to those skilled in the art.
Thus, climatic zone I is a temperate climate; climatic zone II is a
subtropical and mediterranean climate; climatic zone III is hot and
dry; and climatic zone IV is hot and humid.
[0008] According to the present invention there is provided a
liquid composition (e.g. a liquid pharmaceutical composition)
comprising carbetocin or a pharmaceutically active salt thereof;
wherein the pH of the composition is from 5.0 to 6.0. The pH of the
composition may be from 5.1 to 6.0, for example from 5.2 to 6, for
example from 5.26 to 6. The pH of the composition may be from 5.15
to 5.75, for example from 5.2 to 5.65. The pH of the composition
may be from 5.26 to 5.8, for example from 5.26 to 5.75, for example
from 5.26 to 5.7, for example from 5.26 to 5.65, for example 5.4 to
5.65. Preferably, the liquid composition (e.g. a liquid
pharmaceutical composition) comprises carbetocin.
[0009] Preferably the composition is an aqueous composition (e.g.
an aqueous pharmaceutical composition) comprising carbetocin or a
pharmaceutically active salt thereof; wherein the pH of the
composition is from 5.0 to 6.0. The pH of the composition may be
from 5.1 to 6.0, for example from 5.2 to 6, for example from 5.26
to 6. The pH of the composition may be from 5.15 to 5.75, for
example from 5.2 to 5.65. The pH of the composition may be from
5.26 to 5.8, for example from 5.26 to 5.75, for example from 5.26
to 5.7, for example from 5.26 to 5.65, for example 5.4 to 5.65. It
will be appreciated that the composition of the invention is
preferably an aqueous solution. Although water (e.g. water for
injection or WFI) is the preferred solvent, other solvents
(mixtures of water with other pharmaceutically acceptable solvents,
pharmaceutically acceptable alcohols, etc.) may be used.
[0010] The Applicants have found (Examples 1 to 3) that a
composition, e.g. a pharmaceutical composition, comprising
carbetocin or pharmaceutically active salt thereof, and having pH
within a defined specific pH range, may be stored at room
temperature (e.g. at 25.degree. C. and 60% relative humidity) for a
sustained period (e.g. up to 2 years). The composition may also
have a long-term temperature stability at 30.degree. C. and
40.degree. C. and relative humidity up to 75%, and therefore be
suitable for use in zone III/IV regions without requirement for
refrigeration.
[0011] The composition may include (comprise) a buffering agent,
for example a pharmaceutically acceptable buffering agent. Herein,
the term buffering agent is an agent which is capable of driving an
acidic or basic solution to a certain pH state, and then preventing
a change from that state; in other words a buffering agent is an
agent which is added to an already acidic or basic solution to
modify the pH and then maintain the pH at the modified level.
Generally a buffering agent is a weak acid or a weak base that
would be comprised in a buffer solution, and be responsible for the
buffering action seen in these solutions. The buffering agent may
be, for example, acetic acid, adipic acid, citric acid, maleic
acid, succinic acid or phosphate (e.g. sodium phosphate, e.g.
sodium phosphate dibasic dihydrate). Preferably, the buffering
agent is succinic acid. The composition may include a single
buffering agent (i.e. not include two or more buffering agents).
The composition may include two or more buffering agents (e.g.
citric acid and (e.g. sodium) phosphate.
[0012] In another aspect, the composition may include (comprise) a
buffer solution. Herein, the term buffer or buffer solution means a
solution including a mixture of a weak acid and its conjugate base
or a weak base an its conjugate acid, which has the property that
the pH of the solution changes very little when a small amount of
strong acid or base is added, such that the pH of the buffer
(solution) is maintained. The buffer (solution) may be, for
example, a citrate buffer (solution), comprising citric acid and a
citrate (e.g. sodium citrate); a succinate buffer (solution)
comprising succinic acid and a succinate (e.g. sodium succinate),
an acetate buffer (solution) comprising acetic acid and an acetate
(e.g. sodium acetate); a citrate/phosphate buffer (solution)
comprising citric acid and phosphate; or a phosphate buffer
(solution) comprising e.g. (monosodium) phosphate and its conjugate
base, (disodium phosphate). A preferred buffer is a succinate
buffer. The composition may include a single buffer (i.e. not
include two or more buffers). The composition may include two or
more buffers.
[0013] The Applicants have found that inclusion of succinic acid
buffering agent (or use of a succinate buffer) may provide
effective room temperature stability (e.g. at 25.degree. C. and 60%
relative humidity) while possibly conferring additional
advantages--for example, the use of a succinic acid buffering agent
or succinate buffer may contribute to reduced injection site
reactions and associated pain compared with other buffered
formulations.
[0014] The concentration of carbetocin in the liquid (composition
e.g. aqueous composition) may be from 0.01 to 55 mg/mL, for example
0.01 to 50 mg/mL, for example 0.01 to 10 mg/mL, for example 0.01 to
1.5 mg/mL, preferably 0.05 to 0.5 mg/mL, for example 0.1 mg/mL. The
concentration of carbetocin in the liquid (composition e.g. aqueous
composition) may be, for example, 1 mg/mL, 10 mg/mL, 50 mg/mL
etc.
[0015] The compositions of the invention may further comprise an
anti-oxidant. The anti-oxidant may be any anti-oxidant commonly
used in the art, for example any anti-oxidant approved for use as a
pharmaceutical excipient. For example, the anti-oxidant may be
methionine, EDTA, butylated hydroxy toluene, sodium metabisulfite
etc. Preferably the anti-oxidant is present in an amount of 0.01%
to 10% (w/v), for example 0.05% to 5% (w/v), most preferably 0.08%
to 1% (w/v). Preferably the anti-oxidant is methionine, EDTA, or a
combination of methionine and EDTA. For example, the antioxidant
may be methionine and present in an amount of 0.1% w/v (or 1
mg/mL--see Example 2).
[0016] The composition may further comprise an isotonicity agent.
Isotonicity agents, for example mannitol or NaCl, are well known in
the art. Preferably the isotonicity agent is present in an amount
sufficient to provide an isotonic composition (solution), for
example in an amount of 0.01% to 10% (w/v). Preferably the
isotonicity agent is mannitol. If the isotonicity agent is mannitol
it may be present in an amount of 0.5% to 7.5% (w/v), more
preferably 4.0% to 5.5% (w/v), for example 5.0% (w/v). If the
isotonicity agent is mannitol it may be present in an amount of
0.05% to 7.5% (w/v). If the isotonicity agent is NaCl, it may be
present in an amount of 0.05% to 1.2% (w/v), more preferably 0.08%
to 1% (w/v), for example 0.9% (w/v). The isotonicity agent may be
present in an amount of 0.1 to 100 mg/mL, for example 0.5 to 7
mg/mL, for example 1 to 5 mg/mL. For example, if the isotonicity
agent is mannitol it may be present in an amount of 5 to 75 mg/mL,
for example 40 to 55 mg/mL (see e.g. Table 3a). If the isotonicity
agent is NaCl it may be present in an amount of 0.5 to 12 mg/mL,
for example 8 to 10 mg/mL (see e.g. Table 3b), for example 7.5
mg/mL (see Example 6).
[0017] The composition may be for any route of drug administration,
e.g. oral, rectal, buccal, nasal, vaginal, transdermal (e.g. patch
technology); parenteral, intravenous, intramuscular or subcutaneous
injection; intracisternal, intravaginal, intraperitoneal, local
(powders, ointments or drops) or as a buccal or nasal spray.
Preferably the composition is an injectable composition or
injectable formulation. Injectable formulations can be supplied in
any suitable container, e.g. ampoule, vial, pre-filled syringe,
injection device (e.g. single use injection device such as that
sold under the mark Uniject by Becton Dickinson), injection
cartridge, ampoule, (multi-) dose pen and the like. Preferably the
composition is for intramuscular administration (e.g. intramuscular
injection) or intravenous administration (e.g. IV injection).
[0018] The composition may include an enhancer, an excipient which
enhances the effective dose (e.g. enhances the effective dose
following nasal administration). The enhancer may be any enhancer
commonly used in the art, for example any enhancer approved for use
as a pharmaceutical excipient. The enhancer may be, for example,
methyl-.beta.-cyclodextrin, Polysorbate 80, carboxymethylcellulose
or hydroxypropylmethylcellulose.
[0019] The compositions of the invention may be for use in (or in
the manufacture of medicaments for) the treatment or prevention of
uterine atony. The compositions may be for use in the treatment or
prevention of uterine atony following vaginal delivery of the
infant. The compositions may be for use in the treatment or
prevention of uterine atony following delivery of the infant by
Caesarean section, for example delivery of the infant by Caesarean
section under epidural or spinal anaesthesia. The compositions may
be for use in the treatment or prevention of uterine atony, for
example in a patient who is at risk of developing PPH. The
compositions may be for use in (or in the manufacture of
medicaments for) the treatment or prevention of bleeding (e.g.
excessive bleeding) following vaginal delivery (of the infant). The
compositions of the invention may be for use as a uteronic
formulation. The compositions of the invention may be for (e.g.
routine) administration following vaginal delivery of the
infant.
[0020] According to the present invention in a further aspect there
is provided a method of treatment or prevention of uterine atony
(for example following vaginal delivery of the infant or delivery
of the infant by Caesarean section, or in a patient who is at risk
of developing PPH) or a method of treatment or prevention of
excessive bleeding following vaginal delivery comprising, a step of
administration to a patient in need thereof a composition as set
out above.
[0021] It is preferred that the compositions of the invention do
not include a quaternary amine compound, such as benzalkonium
chloride. It is preferred that the compositions of the invention do
not include a parahydroxybenzoate preservative, or a combination of
parahydroxybenzoate preservative with a cosolvent. It is preferred
that the compositions of the invention have a content of divalent
metal ions of less than 2 mM, for example 0.195 mM or less, for
example 0.1 nM or less. It is preferred that compositions of the
invention do not include a solubilizer. It is preferred that
compositions of the invention do not include
methyl-.beta.-cyclodextrin.
[0022] In another aspect of the invention, there are provided
stabilised formulations of carbetocin or other pharmaceutically
active compounds (e.g. other pharmaceutically active peptides or
pharmaceutically active small molecules). Thus, according to the
invention in a further aspect there is provided a liquid (e.g.
aqueous) composition comprising: a pharmaceutically active compound
or salt thereof; and an anti-oxidant; wherein the pH of the
composition is from 5.0 to 6.0. The pH of the composition may be
from 5.1 to 6.0, for example from 5.2 to 6, for example from 5.26
to 6. The pH of the composition may be from 5.15 to 5.75, for
example from 5.2 to 5.65. The pH of the composition may be from 5.0
to 5.9, for example from 5.1 to 5.9, for example 5.2 to 5.8. The pH
of the composition may be from 5.26 to 5.8, for example from 5.26
to 5.75, for example from 5.26 to 5.7, for example from 5.26 to
5.65, for example 5.4 to 5.65. The pharmaceutically active compound
may be carbetocin. The composition may be for nasal administration.
The pharmaceutically active compound may be a compound having the
formula (I) or solvate or pharmaceutically acceptable salts
thereof:
##STR00001##
[0023] wherein: n is selected from 0, 1 and 2; p is selected from
0, 1, 2, 3, 4, 5 and 6; R.sub.1 is selected from aryl optionally
substituted with at least one OH, F, Cl, Br, alkyl or O-alkyl
substituent; R.sub.2 is selected from R.sub.4, H, alkyl,
cycloalkyl, aryl and 5- and 6-membered heteroaromatic ring systems;
R.sub.3 is selected from H and a covalent bond to R.sub.2, when
R.sub.2 is R.sub.4, to form a ring structure; R.sub.4 is C.sub.1-6
alkylene moiety substituted with at least one O-alkyl, S-alkyl or
OH substituent; W and X are each independently selected from
CH.sub.2 and S, but may not both be CH.sub.2; alkyl is selected
from C.sub.1-6 straight and C.sub.4-8 branched chain alkyl and
optionally has at least one hydroxyl substituent; aryl is selected
from phenyl and mono- or poly-substituted phenyl; with the proviso
that when R.sub.2 is H, p is 1, R.sub.3 is H, n is 1 and W and X
are both S, R.sub.1 is not 4-hydroxyphenyl. In the above and
herein, aryl denotes an aromatic group selected from phenyl and
mono- or polysubstituted phenyl; the substituent moieties, if
present, may be selected from fluorine (F), chlorine (Cl) and
bromine (Br) atoms and alkyl, hydroxy (--OH), alkoxy (--O-alkyl)
and alkylthio (--S-alkyl). Preferably the pharmaceutically active
compound is a compound according to formula (I) above with the
proviso that when R.sub.2 is H, p is 0, R.sub.3 is H, n is 1 and W
and X are both S, R.sub.1 is not 4-hydroxyphenyl. These compounds,
their medical uses and their methods of their preparation are
disclosed in WO2009/122285 (International Patent Application No.
PCT/IB2009/005351) of Ferring B.V.
[0024] Preferably the pharmaceutically active compound is
carba-1-[4-FBzlGly.sup.7]dOT, wherein 4-FBzlGly is
N-(4-fluorobenzyl)glycine. Carba-1-[4-FBzlGly.sup.7]dOT is an
oxytocin analogue also known as FE 202767. FE 202767 has the
structure of formula (II):
##STR00002##
[0025] FE 202767 is a selective oxytocin receptor agonist being
evaluated for clinical use. FE 202767 of formula (II), some medical
uses thereof, and methods of its preparation are disclosed in
WO2009/122285. FE202767 is Example 49 of WO2009/122285
(International Patent Application No. PCT/IB2009/005351) of Ferring
B.V.
[0026] Peptidic oxytocin agonists such as those disclosed in
WO2009/122285 are expected to be delivered by the intranasal route
(i.e. intranasal administration). Intranasal formulations of this
type are generally contained in (and administered using) spray
devices, where the drug remains in solution (e.g. at a
concentration of active compound of 0.05-2 mg/mL) under inert
atmosphere for an extended period of time (up to 2 years). There is
therefore a need for room temperature stable formulations of
oxytocin agonists/analogues such as carba-1-[4-FBzlGly.sup.7]dOT
(FE 202767), e.g. to avoid requirement for refrigeration.
[0027] The composition may comprise a buffering agent, for example
acetic acid, adipic acid, citric acid, maleic acid, succinic acid
or (e.g. sodium) phosphate. The composition may include a single
buffering agent. The composition may include more than one
buffering agent (e.g. may comprise citric acid and (e.g. sodium)
phosphate). The composition may comprise a buffer (solution), for
example, a citrate buffer (solution), comprising citric acid and a
citrate (e.g. sodium citrate); a succinate buffer (solution)
comprising succinic acid and a succinate (e.g. sodium succinate),
an acetate buffer (solution) comprising acetic acid and an acetate
(e.g. sodium acetate); a citrate/phosphate buffer (solution)
comprising citric acid and phosphate; or a phosphate buffer
(solution). It is preferred, however, that if the pharmaceutically
active compound is 1-deamino-8-D-arginine vasopressin
(desmopressin) the buffering agent is not malic acid (and/or the
composition does not include malic acid buffer).
[0028] The concentration of the pharmaceutically acceptable
compound in the liquid (aqueous) composition may be, for example,
0.01 to 5 mg/mL.
[0029] The concentration of carbetocin in the liquid (aqueous)
composition may be from 0.01 to 55 mg/mL, for example 0.01 to 50
mg/mL, for example 0.01 to 10 mg/mL, for example 0.01 to 1.5 mg/mL,
preferably 0.05 to 0.5 mg/mL, for example 0.1 mg/mL. The
concentration of carbetocin in the liquid (composition e.g. aqueous
composition) may be, for example, 1 mg/mL, 10 mg/mL, 50 mg/mL
etc.
[0030] The concentration of the compound of formula (II) in the
liquid (aqueous) composition may be from 0.01 to 4 mg/mL, for
example 0.05 to 2 mg/mL, more preferably 0.1 to 1.4 mg/mL, most
preferably 0.2 to 0.7 mg/mL.
[0031] The compositions of this aspect of the invention comprise an
anti-oxidant. The anti-oxidant may be any anti-oxidant commonly
used in the art, for example any anti-oxidant approved for use as a
pharmaceutical excipient. For example, the anti-oxidant may be
methionine, EDTA, butylated hydroxy toluene, sodium metabisulfite
etc. Preferably the anti-oxidant is present in an amount of 0.01%
to 10% (w/v), for example 0.05% to 5% (w/v), most preferably 0.08%
to 1% (w/v). Preferably the anti-oxidant is methionine, EDTA, or a
combination of methionine and EDTA. In an example, the antioxidant
is methionine and is present in an amount of 0.5% w/v. In an
example, the antioxidant is EDTA and is present in an amount of
0.1% w/v.
[0032] The composition may further comprise an isotonicity agent.
Isotonicity agents, for example mannitol or NaCl, are well known in
the art. Preferably the isotonicity agent is present in an amount
sufficient to provide an isotonic composition (solution), for
example in an amount of 0.01% to 10% (w/v). Preferably the
isotonicity agent is mannitol. If the isotonicity agent is mannitol
it may be present in an amount of 0.5% to 7.5% (w/v), more
preferably 4.0% to 5.5% (w/v), for example 5.0% (w/v). If the
isotonicity agent is mannitol it may be present in an amount of
0.05% to 7.5% (w/v). If the isotonicity agent is NaCl it may be
present in an amount of 0.05% to 1.2% (w/v), more preferably 0.08%
to 1% (w/v), for example 0.9% (w/v). The isotonicity agent may be
present in an amount of 0.1 to 100 mg/mL, for example 0.5 to 7
mg/mL, for example 1 to 5 mg/mL. For example, if the isotonicity
agent is mannitol it may be present in an amount of 5 to 75 mg/mL,
for example 40 to 55 mg/mL. If the isotonicity agent is NaCl it may
be present in an amount of 0.5 to 12 mg/mL, for example 8 to 10
mg/mL.
[0033] The composition may be for any route of drug administration,
e.g. oral, rectal, buccal, nasal, vaginal, transdermal (e.g. patch
technology); parenteral, intravenous, intramuscular or subcutaneous
injection; intracisternal, intravaginal, intraperitoneal, local
(powders, ointments or drops) or as a buccal or nasal spray.
Preferably the composition is for nasal administration, e.g. is a
nasal spray. As indicated above, compositions suitable for
intranasal administration are expected to remain stable in solution
at room temperature for an extended period of time.
[0034] The composition may include an enhancer, an excipient which
enhances the effective dose (e.g. enhances the effective dose
following nasal administration). The enhancer may be any enhancer
commonly used in the art, for example any enhancer approved for use
as a pharmaceutical excipient. The enhancer may be, for example,
methyl-.beta.-cyclodextrin, Polysorbate 80, carboxymethylcellulose
or hydroxypropylmethylcellulose.
[0035] It is preferred that the compositions of the invention do
not include a quaternary amine compound, such as benzalkonium
chloride. It is preferred that the compositions of the invention do
not include a parahydroxybenzoate preservative, or a combination of
parahydroxybenzoate preservative with a cosolvent. It is preferred
that the compositions of the invention have a content of divalent
metal ions of less than 2 mM, for example 0.195 mM or less, for
example 0.1 nM or less. It is preferred that compositions of the
invention do not include a solubilizer. It is preferred that
compositions of the invention do not include
methyl-.beta.-cyclodextrin.
[0036] According to the invention in a further aspect there is
provided a liquid (e.g. aqueous) composition comprising: a
pharmaceutically active compound according to formula (II):
##STR00003##
[0037] or (pharmaceutically acceptable) salt thereof; wherein the
pH of the composition is from 5.0 to 6.0. The pH of the composition
may be from 5.0 to 5.9, for example from 5.1 to 5.9, for example
5.2 to 5.8. The pH of the composition may be from 5.1 to 6.0, for
example from 5.2 to 6, for example from 5.26 to 6. The pH of the
composition may be from 5.15 to 5.75, for example from 5.2 to 5.65.
The pH of the composition may be from 5.26 to 5.8, for example from
5.26 to 5.75, for example from 5.26 to 5.7, for example from 5.26
to 5.65, for example 5.4 to 5.65. Preferably, the liquid (e.g.
aqueous) composition comprises a pharmaceutically active compound
according to formula (II):
##STR00004##
[0038] The composition may be for any route of drug administration,
e.g. oral, rectal, buccal, nasal, vaginal, transdermal (e.g. patch
technology); parenteral, intravenous, intramuscular or subcutaneous
injection; intracisternal, intravaginal, intraperitoneal, local
(powders, ointments or drops) or as a buccal or nasal spray.
Preferably the composition is for nasal administration, e.g. is a
nasal spray. As indicated above, compositions suitable for
intranasal administration are expected to remain stable in solution
at room temperature for an extended period of time. Remarkably, the
applicants have found that compositions of the compound of formula
(II) above may provide such stability at pH values which are
particularly suitable for nasal administration, without requirement
for inclusion of an anti-oxidant (see Example 7).
[0039] The composition may comprise a buffering agent, for example
acetic acid, adipic acid, citric acid, maleic acid, succinic acid
or (e.g. sodium) phosphate. The composition may include a single
buffering agent. The composition may include more than one
buffering agent (e.g. may comprise citric acid and (e.g. sodium)
phosphate). The composition may comprise a buffer (solution), for
example, a citrate buffer (solution), comprising citric acid and a
citrate (e.g. sodium citrate); a succinate buffer (solution)
comprising succinic acid and a succinate (e.g. sodium succinate),
an acetate buffer (solution) comprising acetic acid and an acetate
(e.g. sodium acetate); a citrate/phosphate buffer (solution)
comprising citric acid and phosphate; or a phosphate buffer
(solution).
[0040] Preferably the pharmaceutical composition comprises a
citrate/phosphate buffer and the pH is from 5.1 to 6.0, for example
from 5.2 to 6, for example from 5.26 to 6.
[0041] Preferably the pharmaceutical composition comprises a
succinate buffer or a citrate buffer and the pH is from 5.0 to 5.9,
for example 5.0 to 5.8, for example from 5 to 5.7.
[0042] The concentration of the compound of formula (II) in the
liquid (aqueous) composition may be from 0.01 to 4 mg/mL, for
example 0.05 to 2 mg/mL, more preferably 0.1 to 1.4 mg/mL, most
preferably 0.2 to 0.7 mg/mL.
[0043] The compositions of this aspect of the invention further
comprise an anti-oxidant. The anti-oxidant may be any anti-oxidant
commonly used in the art, for example any anti-oxidant approved for
use as a pharmaceutical excipient. For example, the anti-oxidant
may be methionine, EDTA, butylated hydroxy toluene, sodium
metabisulfite etc. Preferably the anti-oxidant is present in an
amount of 0.01% to 10% (w/v), for example 0.05% to 5% (w/v), most
preferably 0.08% to 1% (w/v). Preferably the anti-oxidant is
methionine, EDTA, or a combination of methionine and EDTA. In an
example, the antioxidant is methionine and is present in an amount
of 0.5% w/v. In an example, the antioxidant is EDTA and is present
in an amount of 0.1% w/v.
[0044] The composition may further comprise an isotonicity agent.
Isotonicity agents, for example mannitol or NaCl, are well known in
the art. Preferably the isotonicity agent is present in an amount
sufficient to provide an isotonic composition (solution), for
example in an amount of 0.01% to 10% (w/v). Preferably the
isotonicity agent is mannitol. If the isotonicity agent is mannitol
it may be present in an amount of 0.5% to 7.5% (w/v), more
preferably 4.0% to 5.5% (w/v), for example 5.0% (w/v). If the
isotonicity agent is mannitol it may be present in an amount of
0.05% to 7.5% (w/v). If the isotonicity agent is NaCl it may be
present in an amount of 0.05% to 1.2% (w/v), more preferably 0.08%
to 1% (w/v), for example 0.9% (w/v). The isotonicity agent may be
present in an amount of 0.1 to 100 mg/mL, for example 0.5 to 7
mg/mL, for example 1 to 5 mg/mL. For example, if the isotonicity
agent is mannitol it may be present in an amount of 5 to 75 mg/mL,
for example 40 to 55 mg/mL. If the isotonicity agent is NaCl it may
be present in an amount of 0.5 to 12 mg/mL, for example 8 to 10
mg/mL.
[0045] It is preferred that the compositions of the invention do
not include a quaternary amine compound, such as benzalkonium
chloride. It is preferred that the compositions of the invention do
not include a parahydroxybenzoate preservative, or a combination of
parahydroxybenzoate preservative with a cosolvent. It is preferred
that the compositions of the invention have a content of divalent
metal ions of less than 2 mM, for example 0.195 mM or less, for
example 0.1 nM or less. It is preferred that compositions of the
invention do not include a solubilizer. It is preferred that
compositions of the invention do not include
methyl-.beta.-cyclodextrin.
[0046] According to the present invention in a further aspect,
there is provided a method of treatment or prevention of uterine
atony [for example, treatment or prevention of uterine atony
following vaginal delivery of the infant, treatment or prevention
of uterine atony following delivery of the infant by Caesarean
section, for example delivery of the infant by Caesarean section
under epidural or spinal anaesthesia, or treatment or prevention of
uterine atony in a patient who is at risk of developing PPH], or a
method of treatment or prevention of bleeding (e.g. excessive
bleeding) following vaginal delivery (of the infant), comprising:
administration to a patient in need thereof a liquid (e.g. aqueous)
pharmaceutical composition comprising carbetocin or a
pharmaceutically active salt thereof; wherein the pH of the
composition is from 5.0 to 6.0. The pH of the composition may be
from 5.1 to 6.0, for example from 5.2 to 6, for example from 5.26
to 6. The pH of the composition may be from 5.15 to 5.75, for
example from 5.2 to 5.65. The pH of the composition may be from
5.26 to 5.8, for example from 5.26 to 5.75, for example from 5.26
to 5.7, for example from 5.26 to 5.65, for example 5.4 to 5.65.
[0047] According to the present invention in a further aspect,
there is provided a kit of parts comprising: a liquid (e.g.
aqueous) pharmaceutical composition comprising carbetocin or a
pharmaceutically active salt thereof wherein the pH of the
composition is from 5.0 to 6.0; and a container [e.g. ampoule,
vial, pre-filled syringe, injection device (e.g. single use
injection device such as that sold under the mark Uniject by Becton
Dickinson), injection cartridge, ampoule, multi-dose pen] for the
composition, optionally with separate injection means (e.g. if
required for administration), optionally with instructions for
administration of the composition. The pH of the composition may be
from 5.1 to 6.0, for example from 5.2 to 6, for example from 5.26
to 6. The pH of the composition may be from 5.15 to 5.75, for
example from 5.2 to 5.65. The pH of the composition may be from
5.26 to 5.8, for example from 5.26 to 5.75, for example from 5.26
to 5.7, for example from 5.26 to 5.65, for example 5.4 to 5.65.
[0048] According to the present invention in a further aspect,
there is provided a kit of parts comprising: a liquid (e.g.
aqueous) pharmaceutical composition comprising a pharmaceutically
active compound (e.g. carbetocin) or salt thereof and optionally an
anti-oxidant, wherein the pH of the composition is from 5.0 to 6.0;
and a container (e.g. vial, pre-filled syringe, injection device
[e.g. single use pre-filled injection device such as that sold
under the mark Uniject by Becton Dickinson), injection cartridge,
ampoule, multi-dose pen] for the composition, optionally with
separate injection means (e.g. if required for administration),
optionally with instructions for administration of the composition.
The pH of the composition may be from 5.1 to 6.0, for example from
5.2 to 6, for example from 5.26 to 6. The pH of the composition may
be from 5.15 to 5.75, for example from 5.2 to 5.65. The pH of the
composition may be from 5.26 to 5.8, for example from 5.26 to 5.75,
for example from 5.26 to 5.7, for example from 5.26 to 5.65, for
example 5.4 to 5.65.
[0049] According to the present invention in a further aspect,
there is provided a method of treatment or prevention of
compromised lactation conditions, labour induction impairment,
uterine atony conditions, excessive bleeding, inflammation, pain,
abdominal pain, back pain, male and female sexual dysfunction,
irritable bowel syndrome (IBS), constipation, gastrointestinal
obstruction, autism, stress, anxiety, depression, anxiety disorder,
surgical blood loss, post-partum haemorrhage, wound healing,
infection, mastitis, placenta delivery impairment, osteoporosis,
and a method for the diagnosis of cancer and placental
insufficiency, comprising: administration to a patient in need
thereof a liquid (e.g. aqueous) pharmaceutical composition
comprising a pharmaceutical compound according to formula (I) or
(II) as defined above or a pharmaceutically active salt thereof;
wherein the pH of the composition is from 5.0 to 6.0. According to
the present invention in a still further aspect, there is provided
a liquid (e.g. aqueous) pharmaceutical composition comprising a
pharmaceutical compound according to formula (I) or (II) as defined
above or a pharmaceutically active salt thereof; wherein the pH of
the composition is from 5.0 to 6.0; for use in (or in the
manufacture of a medicament for) the treatment or prevention of
compromised lactation conditions, labour induction impairment,
uterine atony conditions, excessive bleeding, inflammation, pain,
abdominal pain, back pain, male and female sexual dysfunction,
irritable bowel syndrome (IBS), constipation, gastrointestinal
obstruction, autism, stress, anxiety, depression, anxiety disorder,
surgical blood loss, post-partum haemorrhage, wound healing,
infection, mastitis, placenta delivery impairment, osteoporosis, or
for use in (or in the manufacture of a substance for) the diagnosis
of cancer or placental insufficiency. The pH of the composition may
be from 5.1 to 6.0, for example from 5.2 to 6, for example from
5.26 to 6. The pH of the composition may be from 5.0 to 5.9, for
example from 5.1 to 5.9, for example 5.2 to 5.8. The pH of the
composition may be from 5.15 to 5.75, for example from 5.2 to 5.65.
The pH of the composition may be from 5.26 to 5.8, for example from
5.26 to 5.75, for example from 5.26 to 5.7, for example from 5.26
to 5.65, for example 5.4 to 5.65.
[0050] According to the present invention in a further aspect,
there is provided a kit of parts comprising: a liquid (e.g.
aqueous) pharmaceutical composition comprising carbetocin or a
pharmaceutically active compound according to formula (II):
##STR00005##
or a pharmaceutically active salt thereof wherein the pH of the
composition is from 5.0 to 6.0; and a container [e.g. ampoule,
vial, pre-filled syringe, injection device (e.g. single use
injection device such as that sold under the mark Uniject by Becton
Dickinson), injection cartridge, ampoule, multi-dose pen] for the
composition, optionally with separate injection means (e.g. if
required for administration), optionally with instructions for
administration of the composition. The pH of the composition may be
from 5.0 to 5.9, for example from 5.1 to 5.9, for example 5.2 to
5.8. The pH of the composition may be from 5.1 to 6.0, for example
from 5.2 to 6, for example from 5.26 to 6. The pH of the
composition may be from 5.15 to 5.75, for example from 5.2 to 5.65.
The pH of the composition may be from 5.26 to 5.8, for example from
5.26 to 5.75, for example from 5.26 to 5.7, for example from 5.26
to 5.65, for example 5.4 to 5.65.
[0051] According to the present invention in a further aspect,
there is provided a kit of parts comprising: a liquid (e.g.
aqueous) pharmaceutical composition comprising a pharmaceutically
active compound (e.g. carbetocin, a compound of formula (I) or (II)
above) or salt thereof and optionally an anti-oxidant, wherein the
pH of the composition is from 5.0 to 6.0; and a container (e.g.
vial, pre-filled syringe, injection device [e.g. single use
pre-filled injection device such as that sold under the mark
Uniject by Becton Dickinson), injection cartridge, ampoule,
multi-dose pen] for the composition, optionally with separate
injection means (e.g. if required for administration), optionally
with instructions for administration of the composition. The pH of
the composition may be from 5.1 to 6.0, for example from 5.2 to 6,
for example from 5.26 to 6. The pH of the composition may be from
5.0 to 5.9, for example from 5.1 to 5.9, for example 5.2 to 5.8.
The pH of the composition may be from 5.15 to 5.75, for example
from 5.2 to 5.65. The pH of the composition may be from 5.26 to
5.8, for example from 5.26 to 5.75, for example from 5.26 to 5.7,
for example from 5.26 to 5.65, for example 5.4 to 5.65.
DETAILED DESCRIPTION OF THE INVENTION
[0052] The present invention will now be illustrated with reference
to the attached drawings in which:
[0053] FIG. 1 shows a UPLC chromatogram of an impurity mix of
carbetocin and degradation products;
[0054] FIG. 1a shows the chemical formulae of carbetocin and
degradation products;
[0055] FIG. 2 shows the content of degradation (hydrolysis) product
[Gly.sup.9OH]carbetocin (see FIG. 1a) in the antioxidant study
samples as a function of time (constant pH);
[0056] FIG. 3 shows the content of degradation (oxidation) product
sulfoxide II carbetocin (see FIG. 1a) in the antioxidant study
samples as a function of time (constant pH);
[0057] FIG. 4 shows individual degradation products at different pH
(pH study, constant antioxidant);
[0058] FIG. 5 shows the sum of degradation products at different pH
(pH study, constant antioxidant),
[0059] FIG. 6 shows the Stability of FE 202767 in selected buffers;
and
[0060] FIG. 7 shows the sum of impurities of various carbetocin
formulations after 12 months at 40.degree. C. and 75% relative
humidity (R.H.), as described in Experiment 3A.
ANALYTICAL METHOD
[0061] This is the analytical method for the carbetocin examples
(Examples 1 to 6), below.
[0062] All solutions were analysed on a Waters Acquity UPLC
(Ultra-high Pressure Liquid chromatography) system using isocratic
conditions. The Mobile phase was 20% acetonitrile (JT Baker, Ultra
Gradient Grade) in 5 mM unbuffered ammonium acetate (Fluka, Ultra
.gtoreq.99.0%). The column was a Waters Acquity UPLC BEH Shield
RP18, 2.1*100 mm, 1.7 .mu.m (Flow: 0.5 ml/min, Column Temp:
50.degree. C.). The injection volume was 20 .mu.l. Detection was
performed by UV at 220 nm. The different impurities were evaluated
as area % of total area.
[0063] FIG. 1 shows a chromatogram of an impurity mix of carbetocin
and its degradation products. The solution contained, carbetocin,
the hydrolysis products [Gly.sup.9OH], [Asp.sup.5] and
[Glu.sup.4]carbetocin, the oxidation products sulfoxide I and
sulfoxide II-carbetocin, the alkaline degradation products [.beta.
Asp.sup.5] and [D-Asn.sup.5]carbetocin and the synthesis related
impurity [D-Cys.sup.8]carbetocin. The chemical formulae of
carbetocin and the degradation products (hydrolysis products,
oxidation products and alkaline degradation products) are shown in
FIG. 1a. The "[Glu.sup.4]carbetocin" type nomenclature is well
known in the art. The resolutions between all peaks were
.gtoreq.2.0.
Example 1: Formulation Antioxidant Study (Constant pH)
[0064] 5.0 grams of D(-)-Mannitol (Ph Eur, Prolabo) was dissolved
in 1000 ml of milliQ-water. This solution was adjusted with acetic
acid (Ph.Eur., Merck) to pH 5.2. This solution was then divided
into four 200 ml aliquots. To aliquot 1, 0.2 gram of EDTA disodium,
dihydrate (Fluke) was added and dissolved. To aliquot 2, 1.0 gram
of L-methionine (Sigma, non-animal source) was added and dissolved.
To aliquot 3, 0.2 gram of EDTA disodium, dihydrate and 1.0 g of
L-methionine was added and dissolved. Nothing was added to aliquot
4. The pH of aliquots 1-3 was adjusted with acetic acid to pH
5.2.+-.0.1. 1 mg of carbetocin (Polypeptide Laboratories) was
transferred to four 10 ml volumetric flasks. Aliquots 1-4 were used
to dissolve the substance and for dilution to volume (0.1 mg/ml
carbetocin). The solutions were transferred to 25 ml blue cap
flasks, and placed in a cabinet at 40.degree. C. and 75% RH. A
sample of the current PABAL.RTM. formulation, pH 3.9 (measured),
was placed in the same cabinet for comparison.
[0065] The solutions were analysed after 2, 6, 12, 22 and 33 weeks
at 40.degree. C. The largest impurities of this study was found to
be the hydrolysis product [Gly.sup.9OH]carbetocin and the oxidation
product [sulfoxide II]carbetocin. The pH in this study (pH 5.2) was
not high enough to start any alkaline degradation of carbetocin.
The content % (w/w) of the major impurity formed by hydrolysis,
[Gly.sup.9OH]carbetocin, and by oxidation, sulfoxide II-carbetocin,
are shown in FIGS. 2 and 3. The product specification allowed for
each impurity is also plotted in each figure as a reference. Thus
it can be seen from FIG. 2 that if the concentration of
[Gly.sup.9OH] carbetocin increases above 1.5% the sample is "out of
specification", that is the sample has degraded such that is no
longer suitable for administration.
[0066] As shown in FIG. 2, the antioxidant study (constant pH)
showed that the formation of hydrolysis products, mainly
[Gly.sup.9OH]carbetocin, was very fast in the current PABAL
formulation (pH 3.9). With regard to the content of
[Gly.sup.9OH]carbetocin the current formulation was quickly
out-of-specification (>1.5%), after 6 weeks at 40.degree. C. All
formulations at pH 5.2 were well below the specification limit
after 33 weeks at 40.degree. C. (0.4-0.6%), indicating that these
formulations are stable for at least 6 months at 40.degree. C. and
75% RH, which is generally accepted to indicate a likely stability
for at least 24 months at 25.degree. C. and 60% RH (i.e. a stable
RT formulation) The results applied for all three hydrolysis
products. In FIG. 3, the addition of antioxidant was shown to be
very effective in slowing down the oxidation of carbetocin, despite
the increased pH of the formulations. The formulation at pH 5.2
that did not contain any additives was out-of-specification
(>0.8%) with regard to the content of sulfoxide II-carbetocin
after approx 20 weeks at 40.degree. C. The formulations containing
methionine or EDTA were all well below the specification limit
after 33 weeks (0.2-0.4%). The formulation containing a combination
of EDTA and methionine did not show any increase of oxidation
products at all, compared to the levels found in the substance
batch. Due to the low pH, the current formulation was not prone to
degradation by oxidation (pH 3.9). The results are shown in
numerical form in the following table (Table 1).
TABLE-US-00001 TABLE 1 Individual and sum of degradation products
(%) after 33 weeks at 40.degree. C. (constant pH). Sulfoxide D- Sum
of Formulation Gly.sup.9OH Asp.sup.5 Glu.sup.4 Sulfoxide I II
.beta.Asp5 Asn.sup.5 impurities Current formulation 6.43 1.15 5.41
0.42 0.38 0.13 0.15 16.4 Mannitol pH 5.2 0.53 0.14 0.42 0.51 0.93
0.13 0.12 3.5 Mannitol pH 0.63 0.20 0.51 0.14 0.25 0.15 0.16 3.2
5.2 + methionine Mannitol pH 0.52 0.15 0.38 0.31 0.39 0.15 0.10 2.9
5.2 + EDTA Mannitol pH 0.44 0.13 0.35 0.10 0.16 0.19 0.17 2.4 5.2 +
methionine + EDTA
[0067] Table 1 includes the sum of degradation products for all
samples, and after 33 weeks the effect of EDTA is clearer. Further,
the sample containing both methionine and EDTA is clearly better
than the others. The evidence points to a linear degradation:
assuming this is indeed the case, the mannitol pH
5.2+methionine+EDTA sample is likely to be in specification--i.e.
suitable for use--for a remarkable 86 weeks at 40.degree. C. This
is based, as is well known in the art, on a linear extrapolation of
the increase in impurity over time to determine when the amount of
impurity would be sufficiently high for the formulation to be "out
of specification".
Example 2: Formulation pH Study (Constant Antioxidant)
[0068] 1.2 grams of succinic acid (Sigma-Aldrich, .gtoreq.99%) and
1.0 g of L-methionine (Sigma, non-animal source) were dissolved in
1000 ml of milliQ water (10 mM). This solution was adjusted, in
aliquots, with diluted NaOH (Ph.Eur., Merck) to pH 4.0, 4.5, 5.2,
5.65, 6.1, 6.5 and 7.0. 55 mg of carbetocin (Polypeptide
Laboratories) was dissolved in 50 ml of milliQ water (1.1 mg/ml).
1.0 ml of the carbetocin solution (1.1 mg/ml) was mixed with 10 ml
of each buffer (0.1 mg/ml carbetocin). 0.55 g of mannitol (5%) was
added to each solution and dissolved. The solutions were
transferred to 15 ml glass vials with screw lid and placed in a
40.degree. C. cabinet at 75% RH.
[0069] The solutions were analysed after 12 and 52 weeks at
40.degree. C. The content of the individual degradation products
and the sum of degradation products is presented in Tables 2a and
2b and FIGS. 4 and 5.
TABLE-US-00002 TABLE 2a Individual and sum of degradation products
(%) at different pH after 12 weeks at 40.degree. C. (pH study,
constant antioxidant). Sample Hydrolysis products Oxidation
products Alkaline impurities pH [Gly.sup.9OH] [Asp.sup.5]
[Glu.sup.4] Sulfoxide I Sulfoxide II [D-Asn.sup.5] [.beta.
Asp.sup.5] Unknown 2.0 min Sum 4.0 2.22 0.50 1.78 N.D N.D N.D. N.D.
N.D. 4.50 4.5 0.84 0.19 0.67 N.D N.D N.D. N.D. N.D. 1.70 5.2 0.23
0.10 0.27 N.D N.D 0.06 0.05 0.09 0.80 5.65 0.15 0.04 0.12 N.D 0.03
0.12 0.08 0.16 0.70 6.1 0.14 0.09 0.13 0.03 0.03 0.30 0.22 0.32
1.26 6.5 0.25 0.13 0.12 0.03 0.03 0.54 0.30 0.48 1.88 7.0 0.52 0.20
0.17 0.02 0.02 1.27 0.47 0.62 3.29
TABLE-US-00003 TABLE 2b Individual and sum of degradation products
(%) at different pH after 52 weeks at 40.degree. C. (pH study,
constant antioxidant). Sample Hydrolysis products Oxidation
products Alkaline impurities pH [Gly.sup.9OH] [Asp.sup.5]
[Glu.sup.4] Sulfoxide I Sulfoxide II [DAsn.sup.5] [.beta.
Asp.sup.5] Unknown 2.0 min Sum 4.0 7.55 1.09 6.23 0.04 0.07 0.00
0.19 0.94 19.3 4.5 2.92 0.57 2.49 0.04 0.05 0.00 0.19 0.40 7.8 5.2
0.78 0.26 0.72 0.05 0.14 0.26 0.25 0.22 3.6 5.65 0.48 0.22 0.52
0.02 0.02 0.45 0.40 0.40 3.8 6.1 0.56 0.35 0.34 0.08 0.11 1.06 0.73
0.77 5.9 6.5 0.88 0.49 0.32 0.03 0.06 2.30 1.25 1.31 8.8 7.0 1.53
0.71 0.43 0.05 0.04 4.10 1.68 1.65 12.9
[0070] As discussed below with reference to Example 3a, the
specification limit for sum of impurities (for the current
PABAL.RTM. formulation) is .ltoreq.5%. As can be seen from Table 2b
("Sum"), the Samples at pH 5.2 and 5.65 (examples of the invention)
are still within specification after 52 weeks (1 year) at
40.degree. C., while all other samples are out of specification
after 52 weeks (1 year) at 40.degree. C.
[0071] The results of the pH study (FIGS. 4, 5) confirmed the
observations of the antioxidant study. The formation of hydrolysis
products ([Gly.sup.9OH], [Asp.sup.5] and [Glu.sup.4]carbetocin) was
effectively reduced by an increase in pH from pH 4.0 to about pH
5.65. At higher pH values (pH 6.1-7.0) the content of hydrolysis
products was again increased. The effectiveness of the antioxidant
at about 1 mg/ml concentration was also confirmed (in the
antioxidant study the concentration of antioxidant was 5 mg/ml).
However, if oxidation is limited in the drug or drug solution (e.g.
if the drug or drug solution is not prone to oxidation), the amount
of antioxidant may be reduced, or use of antioxidant may not be
necessary. Due to the antioxidant, the oxidation of carbetocin was
negligible, regardless of pH. The upper pH limit of an optimised
formulation was instead limited by the alkaline degradation of
carbetocin. The main impurity by alkaline degradation was
[D-Asn.sup.5]carbetocin, which was rapidly increased at pH values
above pH 6.1. The formation of two other minor impurities at high
pH was also observed, [.beta. Asp.sup.5]carbetocin and one unknown
impurity eluting early in the chromatogram (tR: 2.0 min).
[0072] The U-shape of the pH vs. sum of degradation products curve
illustrates the stability plateau of carbetocin at pH 5.0-6.0. At
pH 5.2 the sum of degradation products was found to be only 16% of
the sum of degradation products at pH 4.0 (current formulation).
The optimal pH was found to be somewhere between pH 5.1 to 6, for
example between around pH 5.2 and 5.65.
[0073] Examples 1 and 2 give a very strong indication that
formulations of the invention are room temperature stable for up to
two years.
Example 3: Formulation Study of Isotonicity Agents, NaCl Vs.
Mannitol, at 30.degree. C., 40.degree. C.
[0074] 4.22 grams of citric acid monohydrate (Merck, pro analysi)
was dissolved in 2000 ml of milliQ water (20 mM). This solution was
divided into ten 200 ml aliquots. 1.8 g of sodium chloride (Merck,
pro analysi) was added to five of the flasks, to the other five
flasks 10 g of mannitol (VWR, Ph Eur) was added. According to an
experimental design, 0.2, 0.6 or 1.0 g of L-methionine (Sigma,
non-animal source) was added and the pH was adjusted with 1% NaOH
(Merck, pro analysi) to pH 5.2, 5.65 or 6.1, see Table 3a and 3b. 2
mg of Carbetocin (Polypeptide Laboratories,) was transferred to
twelve 20 ml volumetric flasks and the substance was dissolved in
each buffer (0.1 mg/ml carbetocin). The samples containing 3 mg/ml
methionine were prepared in duplicate, see Table 3a and 3b.
[0075] Two mL of each solution was transferred to LC-vials and
placed in a 30.degree. C./75% RH cabinet. The remaining solutions
were transferred to 25 ml blue cap flasks and placed in a
40.degree. C./75% R.H. cabinet. The level of impurities after 25
weeks in 30.degree. C./75% R.H are shown in the following Tables 3a
and 3b.
TABLE-US-00004 TABLE 3a Sulfoxide Unknown D- D- Formulation
Gly.sup.9OH Asp.sup.5 Glu.sup.4 Sulfoxide I II 2 min
.beta.Asp.sup.5 Asn.sup.5 Cys.sup.6 Sum** Mannitol, pH 0.18 0.04
0.18 0.04 0.03 N.D. N.D. N.D. 0.09 0.80 5.2, 1 mg/mL methionine
Mannitol, pH 0.07 0.03 0.07 0.03 N.D. 0.17 0.15 0.01 0.12 0.95 6.1,
1 mg/mL methionine Mannitol, pH 0.10 0.03 0.08 0.04 0.05 0.09 0.07
0.05 0.12 0.78 5.65, 3 mg/mL methionine, sample 1 Mannitol, pH 0.09
0.02 0.11 0.03 N.D. 0.08 0.04 N.D. 0.11 0.63 5.65, 3 mg/mL
methionine, sample 2 Mannitol, pH 0.17 0.03 0.20 0.02 N.D. N.D.
0.04 N.D. 0.11 0.75 5.2, 5 mg/mL methionine Mannitol, pH 0.10 0.05
0.05 0.04 N.D. 0.23 0.18 0.11 0.12 1.11 6.1, 5 mg/mL methionine
Years to OOS* 8 12 7 "infinity" "infinity" 6 12 "infinity" N/A 4.2
for 3 mg/mL, pH 5.65 sample 2. *Out of specification **of
degradation products
TABLE-US-00005 TABLE 3b (NaCl) Sulfoxide Unknown Beta D- D-
Formulation Gly.sup.9OH Asp.sup.5 Glu.sup.4 Sulfoxide I II 2 min
Asp.sup.5 Asn.sup.5 Cys.sup.6 Sum** NaCl, pH 5.2, 0.21 0.06 0.15
0.02 0.03 ND 0.02 0.12 0.82 1 mg/mL methionine NaCl, pH 6.1, 0.08
0.05 0.04 0.04 0.04 0.2 0.12 0.12 0.11 0.99 1 mg/mL methionine
NaCl, pH 5.65, 0.09 0.03 0.07 0.01 ND 0.12 0.04 0.05 0.11 0.74 3
mg/mL methionine sample 1 NaCl, pH 5.65, 0.09 0.04 0.08 0.02 0.03
0.10 0.05 0.14 0.11 0.78 3 mg/mL methionine sample 2 NaCl, pH 5.2,
0.17 0.04 0.15 0.02 0.04 ND 0.05 0.14 0.86 5 mg/mL methionine NaCl,
pH 6.1, 0.09 0.04 0.06 0.03 ND 0.23 0.17 0.14 0.11 1.11 5 mg/mL
methionine **of degradation products
[0076] Tables 3a and Tables 3b show that there is very little
degradation in all samples. This level of degradation corresponds
to that seen after 6 weeks at 40.degree. C.
[0077] The results indicate that the best samples are likely to be
stable for 5 years at 30.degree. C. As seen in Table 3a, rows 5 and
8, results for the methionine 3 mg/mL, pH 5.65 sample 2 indicate
that this sample would remain in specification for more than 4
years at 30.degree. C. and 75% RH. This is based, as is well known
in the art, on a linear extrapolation of the increase in impurity
over time to determine when the amount of impurity would be
sufficiently high for the formulation to be "out of specification"
(OOS). It was also found that the optimum pH at 30.degree. C. is
higher than at 40.degree. C. (results not shown). The differences
are small but pH 5.65 is slightly superior to pH 5.2 at 30.degree.
C. (vice versa at 40.degree. C.). These results indicate there is a
good margin for obtaining a climate zone III/IV stable
formulation.
[0078] The applicants found that increase in methionine leads to
more degradation, mainly by increase of [BetaAsp5]carbetocin. A
concentration of about 1 mg/ml appears to be sufficient to provide
effective stabilisation without significant degradation.
Experiment 3a--the Stability of Carbetocin at Different pH and
Using Different Antioxidants
[0079] This study was designed to give a broader picture of the
stability of carbetocin at different pH and using different
antioxidants.
[0080] 1.2 grams of succinic acid (Sigma-Aldrich, 99%) was
dissolved in 1000 ml of milliQ water (10 mM). This solution was
adjusted, in aliquots, with diluted NaOH (Ph.Eur., Merck) to pH
4.0, 4.5, 5.2, 5.65, 6.1, 6.5 and 7.0. 55 mg of carbetocin
(Polypeptide Laboratories, Strasbourg) was dissolved in 50 ml of
milliQ water (1.1 mg/ml). 1.0 ml of the carbetocin solution (1.1
mg/ml) was mixed with 10 ml of each buffer (0.1 mg/ml carbetocin).
0.55 g of mannitol (5%) was added to each solution and dissolved.
The solutions were transferred to 15 ml glass vials with screw lid
and placed in the 40.degree. C./75% R.H. cabinet.
[0081] The same procedure was repeated; with the exception that 1.0
g of L-methionine (Sigma, non-animal source) was added to the 1000
ml of milliQ water, giving duplicate samples containing 1 mg/ml
methionine at all pH-levels. The solutions were transferred to 15
ml glass vials with screw lid and placed in the 40.degree. C./75%
R.H. cabinet.
[0082] The buffers at pH 5.65, 6.1 and 6.5 were also divided into
aliquots to which EDTA disodium, dihydrate (Fluka) was added. These
samples were stored at 40.degree. C./75% for 12 months before
analysis.
[0083] The sum of impurities after 12 months at 40.degree. C./75%
R.H are shown in FIG. 7. The Figure also shows the "spec limit",
above which the sum of impurities is such that the formulation is
out of specification.
[0084] All formulations at pH 5.2 and pH 5.65 were within
specification after 12 months at 40.degree. C./75% R.H.
[0085] The positive effect of methionine was visible also in this
study. All samples containing methionine showed very low amounts of
oxidation products, regardless of composition and pH. This points
to inclusion of methionine in a robust formulation, where (for
example) metal ion content of the active ingredient carbetocin,
which can vary with production batch and which, if high, may lead
to increased oxidation, will not be a controlled parameter.
[0086] The most stable formulation was the formulation at pH 5.2
containing 1 mg/ml of methionine (results not shown). The parameter
that was closest to the specification limit after 12 months at
40.degree. C./75% R.H. was the sum of impurities (FIG. 7). The
specification limit for sum of impurities (for the current
PABAL.RTM. formulation) is .ltoreq.5% (i.e. 5.5%). It can be
assumed that the degradation is linear over time, and it can
therefore be calculated that the formulation at pH 5.2 containing 1
mg/ml of methionine would be out-of-specification after
approximately 80 weeks at 40.degree. C./75% R.H, based on the
specification for the current PABAL.RTM. formulation.
[0087] A commonly used guide, supported by the Arrhenius equation,
is that the rate of most chemical reactions doubles for every
10.degree. C. increase of temperature. If we apply this
relationship to the formulation at pH 5.2 containing 1 mg/ml of
methionine, the estimated shelf-life of a new formulation will be
160 weeks at 30.degree. C., i.e. slightly more than 3 years, again
based on the specification for the current PABAL.RTM. formulation.
This is likely to be an underestimation, since the reported "sum of
impurities" in this experiment included every peak on the baseline,
including synthesis related impurities and peaks below the
reporting limit (<0.05%). The synthesis related impurities
consist mainly of [DCys.sup.6] and [desGln.sup.4]carbetocin, which
do not increase during storage. The substance batch contained 0.9%
impurities according to the supplier. Thus, it is likely that a
shelf-life of more than 3 years at 30.degree. C./75% R.H. would be
achieved for this formulation.
Example 4--Formulation in Succinate Buffer
[0088] The following preparation and decanting was performed in a
pharmaceutical room under germ free conditions. 47 grams of
mannitol, 1.2 grams of succinic acid buffering agent and 1.0 g of
L-methionine was dissolved in about 900 ml of milliQ water (10 mM).
The pH of the solution was adjusted with 5M NaOH to pH 5.4. The
solution was transferred to a 1000 ml volumetric flask and diluted
to volume with WFI.
[0089] 50 mg of carbetocin (Polypeptide Laboratories) was
transferred to a 500 ml volumetric flask and dissolved and diluted
to volume with the mannitol/succinic acid/methionine buffer pH 5.4.
The solution was filtered through a 0.22 .mu.m filter and filled in
glass vials with rubber stoppers (1.1 ml per vial). Each vial
included an aqueous composition comprising carbetocin (0.1 mg/mL),
and the pH of the composition was 5.4 (i.e. from 5.0 to 6.0). The
aqueous composition also included succinate buffer (succinic acid
buffering agent), methionine (anti-oxidant) and mannitol (isotonic
agent). In a further Example (Example 4A, not shown) a solution was
made up exactly as Example 4 and EDTA (0.1% w/v) added. The
osmolality of the solutions in Example 4 and 4A was found to be
300.+-.20 mOsmol/kg.
[0090] The formulation of Example 4 (and that of Example 4A) is
suitable for injection to a patient with uterine atony.
Example 5--Formulation in Succinate Buffer
[0091] The following preparation and decanting was performed in a
pharmaceutical room under germ free conditions. 1.2 grams of
succinic acid buffering agent (Sigma-Aldrich, 99%) and 1.0 g of
L-methionine (Sigma, non-animal source) were dissolved in 1000 ml
of milliQ water (10 mM) to provide a succinate buffer of pH 5.4,
the pH being adjusted to this value with NaOH solution.
[0092] 0.55 g of mannitol (5%) was dissolved in 10 ml of succinate
buffer. Methionine 0.5% (w/v) was added to the solution and
dissolved. Carbetocin (Polypeptide Laboratories) was dissolved in
the solution so the concentration of carbetocin was 0.1 mg/mL, and
the pH adjusted to 5.4 using NaOH solution. The solution was
divided into 1 mL quantities and sealed in ampoules. Each ampoule
included an aqueous composition comprising carbetocin (0.1 mg/mL),
and the pH of the composition was 5.4 (i.e. from 5.0 to 6.0). The
aqueous composition also included succinate buffer (succinic acid
buffering agent), methionine (anti-oxidant) and mannitol (isotonic
agent). It will be appreciated that the composition may be made
with water for injection (WFI). The formulation of Example 5 is
suitable for injection to a patient with uterine atony.
Example 6--Formulation with Citrate/Phosphate Buffer
[0093] The formulation set out in the following table was made up
by similar methods to those set out in Examples 4 and 5 above.
TABLE-US-00006 TABLE 4 Component Amount per mL Function Carbetocin
10 mg Active ingredient Sodium phosphate dibasic 3.24 mg Buffering
agent dihdrate Citric acid monohydrate 1.43 mg Buffering agent NaCl
7.5 mg Isotonicity agent HCl q.s. adjust to pH 5.5 pH adjustment
NaOH q.s. adjust to pH 5.5 pH adjustment Water for Injection Adjust
to 1 mL Solvent
[0094] The composition is suitable for nasal administration.
[0095] Optionally, an antioxidant (e.g. methionine at a
concentration of 1.0 mg/mL may be included in the formulation). The
anti-oxidant may be any anti-oxidant commonly used in the art.
[0096] Optionally, the composition may include an enhancer. The
enhancer may be any enhancer commonly used in the art, for example
any enhancer approved for use as a pharmaceutical excipient. The
enhancer may be, for example, methyl-.beta.-cyclodextrin,
Polysorbate 80, carboxymethylcellulose or
hydroxypropylmethylcellulose.
Example 7--the Stability of FE 202767 in Citrate and
Citrate-Phosphate Buffers (pH 5.0, 5.5, and 6.0) at 40.degree. C.
for a Six Month Period
[0097] Materials and Methods
[0098] FE 202767 (Ferring) was synthesised by the method set out in
WO2009/122285. FE 202767 was dissolved at a concentration of 0.2
mg/ml in either 25 mM citrate buffer (isotonic to saline) or 25 mM
citrate-phosphate buffer (isotonic with saline) at varying pH (pH
5.0, 5.5, 6.0), by methods known in the art. The solutions were
incubated at 40.degree. C. for 176 days, with samples taken at day
0, 15, 30, 84, and 176.
[0099] Samples were evaluated by HPLC to determine the amount of
intact peptide remaining at the various time points, by methods
well known in the art, comparing the % Area. of the intact peptide
peak on the sampling day vs. % Area on Day 0.
[0100] The HPLC method used an Agilent 1200 instrument. The mobile
phases where HPLC Buffers A (A=0.01% TFA in water) and B (B=0.01%
TFA in [70% v/v acetonitrile and 30% v/v water]) with the gradient
15% B for 1 min, then 15 to 95% B in 30 min, then 95 to 100% B in 3
min, then 100% B for 5 min and 100% B to 15% B in 1 min at flow
rate 0.3 mL/min. The Phenomenex MAX-RP C18, 2.0.times.150 mm, 4
.mu.m, 80 {acute over (.ANG.)} column was at temperature 40 with UV
detection at 210 nm. The injection volume was 10 .mu.L.
[0101] The results are shown in the following Table 5, and on the
attached FIG. 6. In Table 5 and FIG. 6, CP50 is citrate phosphate
buffer at pH 5.0; CP55 is citrate phosphate buffer at pH 5.5; and
CP60 is citrate phosphate buffer at pH 6.0; CT50 is citrate
phosphate buffer at pH 5.0; CT55 is citrate phosphate buffer at pH
5.5; and CT60 is citrate phosphate buffer at pH 6.0.
TABLE-US-00007 TABLE 5 % Intact Peptide Remaining (normalised to
day 0) Day CP50 CP55 CP60 CT50 CT55 CT60 0 100.00 100.00 100.00
100.00 100.00 100.00 6 99.97 99.83 99.65 99.66 99.75 100.00 15
99.66 99.31 99.54 99.51 99.71 99.40 30 99.47 99.30 99.26 99.29 n.a.
99.30 84 98.38 97.44 97.78 98.05 98.61 97.94 176 96.98 95.60 95.48
95.19 97.34 73.36 Notes: % Intact Peptide Remaining expressed
relative to % Area on Day 0. n.a. = data point excluded due to
aberrant peak in HPLC chromatogram. CP = citrate-phosphate buffer;
CT = citrate buffer.
[0102] Conclusion
[0103] FE 202767 showed good stability in citrate-phosphate buffers
in the pH range tested (pH 5.0, 5.5, and 6.0), with >95%
remaining after 176 days in each condition. It was also very stable
(>95% remaining) in citrate buffer at pH 5.0 and 5.5; however,
there was significant degradation after 176 days in pH 6.0 citrate
buffer.
[0104] In general, a formulation suitable for nasal administration
is expected to be of pH between 5.0 and 6.0, include the minimum
number of reagents (e.g. no anti-oxidant). It is also preferred
that the formulation is room temperature stable. Example 7
demonstrates that formulations along the lines above may be
suitable for nasal administration, because they have appropriate pH
and are room temperature stable without requirement for
anti-oxidant or other additives that might adversely affect the
nasal mucosa.
Example 8--the Stability of FE 202767 in Various Buffers at
40.degree. C. for One and Three Months
[0105] Materials and Methods
[0106] The method was similar to Example 7. FE 202767 (Ferring) was
synthesised by the method set out in WO2009/122285. The FE 202767
was dissolved at a concentration of 0.2 mg/ml in either 25 mM
citrate buffer (citric acid/Na citrate), 10 mM acetate buffer
(acetic acetate/Na acetate) or 10 mM succinate buffer (1 mM
succinic acid+NaOH to relevant pH) at varying pH (pH 5.0, 5.2, 5.5,
5.65, 5.8, 6.0), by methods known in the art. As set out in the
table below, the various samples also included isotonicity agent
(NaCl, 7 mg/mL or mannitol 47 mg/mL) to achieve isotonicity. Some
of the samples included oxidant (methionine 1 mg/mL, EDTA 1 mg/mL,
or combination of EDTA 1 mg/mL and methionine 1 mg/mL). Each
formulation (see Table below) was filled in a 10R glass vial sealed
with a rubber stopper and an aluminium cap.
[0107] The solutions were incubated at 40.degree. C. at 75% RH,
with samples taken at day 30 (1 month), and day 90 (3 months).
[0108] Samples were evaluated by HPLC to determine the amount of
intact peptide remaining at the various time points, by methods
well known in the art, comparing the % Area of the intact peptide
peak on the sampling day vs. % Area on Day 0.
[0109] The HPLC method used an Agilent 1100 instrument. The mobile
phases where HPLC Buffers A (A=0.1% TFA in water) and B (B=0.1% TFA
in acetonitrile) with the gradient 20 to 30% B in 40 min, then 30
to 60% B in 15 min, then 60 to 20% B in 1 min and then 20% B for 10
min at flow rate 0.5 mL/min. The Zorbax 300SB C18, 3.0.times.150
mm, 3.5 .mu.m, 300 {acute over (.ANG.)} column was at temperature
25 with UV detection at 214 nm. The injection volume was 15
.mu.L
[0110] The results are shown in the following Table.
TABLE-US-00008 TABLE 6 Initial Peptide Peptide peptide conc. conc.
Sample Isotonicity conc. (mg/mL) (mg/mL) Number Buffer pH agent
Antioxidant (mg/mL) at 30 days at 90 days 1 Citrate 6 NaCl No 0.186
0.187 0.182 2 Citrate 5.65 NaCl No 0.187 0.187 0.182 3 Citrate 5.8
NaCl No 0.187 0.187 0.182 4 Citrate 5 NaCl Methionine 0.187 0.183
0.162 5 Citrate 5.5 NaCl Methionine 0.187 0.187 0.171 6 Citrate 6
NaCl Methionine 0.186 0.187 0.181 7 Citrate 6 NaCl EDTA 0.187 0.188
0.183 8 Citrate 6 NaCl Methionine 0.187 0.187 0.182 and EDTA 8
placebo Citrate 6 NaCl Methionine 0.000 0.000 0.000 and EDTA 9
Citrate 5 Mannitol No 0.187 0.186 0.173 10 Succinate 6 Mannitol No
0.188 0.186 0.180 11 Succinate 5 NaCl No 0.186 0.187 0.183 12
Succinate 5.2 NaCl No 0.187 0.188 0.184 13 Succinate 5.65 NaCl No
0.187 0.188 0.183 14 Succinate 6 NaCl No 0.187 0.187 0.182 15
Succinate 5 NaCl Methionine 0.187 0.186 0.182 16 Succinate 5.2 NaCl
Methionine 0.187 0.185 0.182 17 Succinate 5.65 NaCl Methionine
0.187 0.186 0.180 18 Succinate 6 NaCl Methionine 0.187 0.188 0.181
19 Succinate 5 Mannitol No 0.188 0.186 0.176 20 Succinate 6
Mannitol No 0.187 0.099 0.124 21 Succinate 5 Mannitol Methionine
0.187 0.180 0.012 21 placebo Succinate 5 Mannitol Methionine 0.000
0.000 0.000 22 Succinate 6 Mannitol Methionine 0.188 0.023 0.174 23
Acetate 5.2 NaCl No 0.186 0.186 0.183 24 Acetate 5.65 NaCl No 0.187
0.187 0.184 24 placebo Acetate 5.65 NaCl No 0.000 0.000 0.000
[0111] Conclusion
[0112] FE 202767 showed good stability in citrate and acetate
buffers in the pH range tested after 30 days in each condition. It
was also very stable in succinate buffer at pH 5.0 to 5.65;
however, there was significant degradation after 30 days in some pH
6.0 succinate samples (sample 20, 22). The presence or absence of
antioxidant seemed unimportant on a 30 day timescale.
[0113] FE 202767 also showed good stability in citrate and acetate
buffers in the pH range tested after 90 days in each condition,
with the best results being shown at the upper end of the pH range
(e.g. between pH 5.5 and 6, see samples 1 to 6). It was also stable
in succinate buffer at pH 5.0 to 5.65 after 90 days. The 30 and 90
day results for samples 21 and 22 suggest a mix up in analysis.
[0114] Again, the presence or absence of antioxidant seemed
unimportant on a 90 day timescale.
[0115] The results indicate that NaCl is a better isotonicity agent
than mannitol.
[0116] As indicated above, a formulation suitable for nasal
administration is expected to be of pH between 5.0 and 6.0, include
the minimum number of reagents (e.g. no anti-oxidant). It is also
preferred that the formulation is room temperature stable. Example
8 demonstrates that formulations along the lines above may be
suitable for nasal administration, because they have appropriate pH
and are room temperature stable without requirement for
anti-oxidant or other additives that might adversely affect the
nasal mucosa.
Example 9--Formulation of FE 202767 with Citrate/Phosphate
Buffer
[0117] FE 202767 (Ferring) was synthesised by the method set out in
WO2009/122285. The formulation set out in the following table was
made up by similar methods to those set out in Examples 4 and 5
above.
TABLE-US-00009 TABLE 7 Component Amount per mL Function
carba-1-[4-FBzlGly7]dOT 0.7 mg Active ingredient (FE 202767) Sodium
phosphate dibasic 3.24 mg Buffering agent dihdrate Citric acid
monohydrate 1.43 mg Buffering agent NaCl 7.5 mg Isotonicity agent
HCl q.s. adjust to pH 5.5 pH adjustment NaOH q.s. adjust to pH 5.5
pH adjustment Water for Injection Adjust to 1 mL Solvent
The composition is suitable for nasal administration.
[0118] Optionally, an antioxidant (e.g. methionine at a
concentration of 1.0 mg/mL may be included in the formulation).
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