Field of Invention
The present invention relates to a stable salicylate compound composition, e.g. aspirin
composition, in liquid form, in particular which is suitable for oral use, intravenous
administration or inhalation.
Background
Aspirin (acetylsalicylic acid) has been widely used as an analgesic to relieve minor
aches and pains, as an antipyretic to reduce fever, and as an anti-inflammatory
medication. Aspirin also has a 'COX' mediated antiplatelet effect by inhibiting the
production of thromboxane, which under normal circumstances binds platelet
molecules together to create a patch over damaged walls of blood vessels. Because
the platelet patch can become too large and also block blood flow, locally and
downstream, aspirin is also used long-term, at low doses, to help prevent heart
attacks, strokes, and blood clot formation in people at high risk of developing blood
clots. Also, low doses of aspirin may be given immediately after a heart attack to
reduce the risk of another heart attack or the death of cardiac tissue. There is also
growing evidence that aspirin can effectively prevent and slow the growth of cancers,
in particular gastrointestinal cancers.
For example, Garcia-Albenix et al. (incorporated herein by reference) demonstrated
that daily ingestion of aspirin over an 8-10 year period, at any dose, reduced the risk of
colorectal cancer ('CRC') by an impressive 24% and CRC-associated mortality by an
even more impressive 35%.
In addition, Lan et al. ["Antitumor effect of aspirin in glioblastoma cells by modulation of
b-catenin/T-cell factor-mediated transcriptional activity", J Neurosurg, 201 1, Vol. 15,
pp. 780-788, incorporated herein by reference] found that aspirin is a potent
antitumour agent which exerts its antineoplastic action by inhibition of the
b-catenin/TCF signalling pathway in glioma cells.
Other studies have also shown aspirin to be a potent antitumour agent in glioma cell
lines [M. W. Brown, 'Characterisation Of The Effects Of Chronic Aspirin Treatment On
The Viability And Proliferation Of Stage 4 Glioblastoma Cells', Diffusion: the UCLan
Journal of Undergraduate Research, Vol. 6, Issue 2, December 2013; Aas et al.,
'Growth inhibition of rat glioma cells in vitro and in vivo by aspirin', Journal of Neuro-
Oncology, 1995, Vol. 24, Issue 2, pp. 171-180; Ning e a/., Overexpression of S100A9
in human glioma and in-vitro inhibition by aspirin', European Journal of Cancer
Prevention, 2013, Vol. 22, Issue 6, pp. 585-595; Hwang e a/., 'Effect of aspirin and
indomethacin on prostaglandin E2 synthesis in C6 glioma cells', Kaohsiung J Med Sci,
2004, Vol. 20, pp. 1-5; Okada er at., 'Integration of epidemiology, immunobiology, and
transiationai research for brain tumors', Ann N Y Acad Sci, 2013, Vol. 1284, pp. 17-23;
all incorporated herein by reference].
Aspirin has often been called the 'wonder drug' or 'miracle cure'. For the last quarter
of a century it has been universally accepted that this Willow tree bark derivative is of
great benefit in cardiovascular disease prevention and cerebrovascular disease
prevention. Over the last decade there has been a growing 'evidence based'
realisation that it can prevent cancer (e.g. gastrointestinal or "Gl" tract cancer). This
anti-cancer effect has been shown to reduce cancer mortality by around 20% (Rothwell
et ai, 'Effect of daily aspirin on long-term risk of death due to cancer: analysis of
individual patient data from randomised trials', Lancet, 201 , Vol. 377, pp. 31-41 ,
incorporated herein by reference).
The main drawback of aspirin is its ability to cause gastric mucosal damage, which is
exacerbated by the presence of particulate aspirin. Lan e a/ (supra) identified these
side-effects as the limiting factor in the treatment of glioma with aspirin. Fully
solubilised, particulate free aspirin would be expected to reduce the local cytotoxic
irritant effect of aspirin.
All current 'stable' aspirin formulations, whether they are called dispersible, soluble or
effervescent, are simply dispersible particle formats, which will settle out into
particulate form soon after being dissolved.
K. D. Rainsford ('Aspirin and Related Drugs', 2004, incorporated herein by reference)
discusses the first definitive evidence of the direct gastric irritant actions of aspirin
particles on the gastric mucosa. This was shown in a variety of studies in the 930s,
which showed that particles of unsolubilised aspirin embedded in the gastric mucosa or
surrounding ulcer sites (Douthwaite and Lintott, 'Gastroscopic observations of the
gastric mucosa after use of aspirin', Lancet, 1938, Vol. 232, pp. 1222-1225; Hurst and
Lintott, 'Aspirin as a cause of hematemesis', Guy's Hosp Rep, 1939, p. 173, both
incorporated herein by reference). Further confirmation of this effect is seen in the
image taken by D. J. Levy ('An aspirin tablet and gastric ulcer', N Engl J Med, 2000,
Vol. 343, No. 12, p. 863, incorporated herein by reference).
Rainsford et al. ('Electronmicroscopic observations on the effects of orally
administered aspirin and aspirin-bicarbonate mixtures on the development of gastric
mucosal damage in the rat', Gut, 1975, Vol. 16(7), pp. 514-527, incorporated herein by
reference) noted that "the absence of particles of the drug in the aspirin and
bicarbonate mixtures is clearly a major factor. A consequence of this is that there
would be an increase of the spread of the drug over the mucosal and gastric pit,
thereby reducing the amount of drug in concentrated particles which seems important
in the development of focal damage and development of gastric mucosal damage".
This is further supported by three further studies by Jaiswal et al. (IUPHAR 9th Int.
Conference of Pharmacology, 1984, incorporated herein by reference), Liversage et al.
('Drug particle size reduction for decreasing gastric irritation and enhanced absorption
of Naproxen in rats', Int J Pharm, 1995, Vol. 125(2), pp. 309-313, incorporated herein
by reference) and Gyory et al. ('Effect of particle size on aspirin-induced
gastrointestinal bleeding', Lancet, Vol. 292, No. 7563, pp. 300-302, incorporated
herein by reference). These all clearly show that there is a definite correlation between
the size of the aspirin particle and the severity of the gastrointestinal irritation.
M. I . Grossman e a/. ('Fecal Blood Loss Produced By Oral And Intravenous
Administration Of Various Salicylates', Gastroenterology, 1961 , Vol. 40, pp. 383-388,
incorporated herein by reference) firmly supports the aspirin 'particle effect' as the
cause of Gl irritation. Grossman suggests that in order for intravenous aspirin to
cause Gl bleeding, there must be pre-existing damage. He states that "the gun must
be loaded in order for an explosion to occur when salicylates pull the trigger".
This is further supported by a series of seven studies, all looking at the effect of
intravenous, subcutaneous or intrajejunal aspirin on the gastric mucosa. Each of these
notes that the aspirin given at varying doses (some of which are extremely high) has a
significant effect on systemic COX and prostaglandin production. In each case it is
stated that despite having this systemic effect the only route of administration that
causes gastrointestinal problems is where there is direct contact with the gastric
mucosa. They all conclude that the effects seen are therefore due to direct contact
'particle' effect rather than 'COX' mediated systemic effects. The systemic COX effect
is not a factor causing the Gl side-effects of aspirin but rather exacerbates any
bleeding from a pre-existing lesion, due to the COX-mediated anti-platelet effect.
[Kevin et a/., 'Acute effect of systemic aspirin on gastric mucosa in man', Digestive
Diseases and Sciences, 1980, Vol. 25, Issue 2, pp 97-99; Cooke et al., 'Failure of
intravenous aspirin to increase gastrointestinal blood loss', British medical journal,
1969; Vol. 3(5666), pp. 330-332; Wallace et al., 'Adaptation of rat gastric mucosa to
aspirin requires mucosal contact' Am J Physiol, 1995, Vol. 268, G 134-8; Cryer et al.
'Effects of low dose daily aspirin therapy on gastric, duodenal and rectal prostaglandin
levels and on mucosal injury in healthy humans', Gastroenterology, 1999, Vol. 17, pp.
17-25; Lichtenberger et al., 'Where is the evidence that cyclooxygenase inhibition is
the primary cause of nonsteroidal anti-inflammatory drug (NSAID)-induced
gastrointestinal injury? Topical injury revisited', Biochemical Pharmacology, 2001 , Vol.
6 1, pp. 631-637; Ligumsky M et al., 'Aspirin can inhibit gastric mucosal cyclooxygenase
without causing lesions in the rat', Gastroenterology, 1983, Vol. 84, pp
756-61 ; and Zhao et al., 'Clinical Research Feasibility of intravenous administration of
aspirin in acute coronary syndrome', Journal of Geriatric Cardiology, 2008, Vol. 5 No.
4 , pp. 212-216, all incorporated herein by reference].
The presence of aspirin particles, in contact with the stomach lining, seems to be
essential for the causation of aspirin Gl irritation. This is not the case for the other
NSAID's where the systemic COX effect is sufficient to induce Gl injury (Mashita et al.,
Oral but not parenteral aspirin upregulates COX-2 expression in rat stomachs: a
relationship between COX-2 expression and PG deficiency', Digestion, 2006, Vol.
73(2-3), pp. 124-32, incorporated herein by reference).
The study by Mashita et al. {ibid.) shows that while both Indomethacin (a 'new' NSAID)
and aspirin both decrease PGE2 when given either orally or subcutaneously, only
Indomethacin caused gastric damage via both administration routes. Aspirin only
caused damage when given orally. They conclude that Aspirin caused damage due to
topical irritation, while Indomethacin caused damage due to its systemic effect. This
highlights the uniqueness of aspirin and distinguishes it from all other NSAIDs.
It is therefore clear that the direct topical irritation of aspirin particles on the gastric
mucosa causes the damage leading to the side effects seen when taking oral aspirin.
It follows that if the particles are removed by producing a stable, fully solubilised liquid
aspirin, the adverse G l side effects will be significantly reduced or even totally
eliminated.
Aspirin is generally only administered orally in solid form, i.e. tablets, granules etc.,
because of the instability of aspirin in solution or liquid form. Aspirin is not soluble and
stable in water, or in any of the common solvents used in oral pharmaceuticals, such
as propylene glycol, ethanol and glycerol. Aspirin is rapidly hydrolyzed to acetic and
salicylic acids which results in a major loss of its pharmacological activity. Thus, the
shelf life of aspirin in such solvents is far too short to permit the development of a
stable aspirin solution suitable for commercial use. Water alone is not the only
contributor to aspirin degradation in solution. Aspirin will also degrade by hydrolysis,
glycolysis and trans-esterification, all of which will be promoted by any pH higher than
about 3.5. There is a need for a stable and soluble aspirin composition in liquid form
which can be taken orally.
Furthermore, other modes of administration suited to liquid drug compositions are at
present not accessible by aspirin due to the instability in solution or liquid form and the
difficulty in achieving complete solubilisation. For example, it is desirable to provide a
stable aspirin composition, preferably containing completely solubilised aspirin, which
may be administered to the bloodstream, e.g. intravenously or intraarterially, or may be
administered to the lungs by inhalation, e.g. of a vapour or aerosol.
One particular advantageous use of a stable liquid aspirin composition is to orally give
to stroke or heart attack victims, e.g. immediately after an attack, where it is often
impossible for the patient to take aspirin in solid form. Other advantages include
convenience, speed and completeness of absorption (increased bioavailability) and
reduced gastric irritation. In addition, for elderly patients or patients suffering from
Dysphagia (swallowing difficulties), it may be impossible or dangerous to administer
solid aspirin orally. A stable liquid composition would address this problem.
Other salicylate compounds present problems associated with achieving a stable liquid
composition in which the salicylate compound is fully solubilised. For example, stable
liquid compositions of Diflunisal (2',4'-difluoro-4-hydroxybiphenyl-3-carboxylic acid),
Triflusal (2-acetyloxy-4-(trifluoromethyl)benzoic acid), Salsalate (2-(2-
hydroxybenzoyl)oxybenzoic acid) and salicylic acid itself (2-hydroxybenzoic acid) may
also be desirable.
WO 91/01761 describes shelf-stable solutions of aspirin in DEET, which may also
include glycerin triacetate as a co-solvent.
EP 0 920 862 describes solutions of aspirin with at least one substance selected from
an ester of an organic acid, a glycerol fatty acid ester, silicon oil and hydrocarbon oil.
US 6 306 843 describes stable aspirin solutions in a non-aqueous organic solvent,
which include a cyclic acid imide and/or a sulfaminic acid.
The compositions described in these documents however do not provide sufficient
stability and/or contain non-GRAS or non-pharmaceutically licensed products which
are unsuitable for human ingestion. There therefore remains a need for stable aspirin
solutions offering extended shelf-life with increased solubility and stability of the
dissolved compound, which are suitable for therapeutic use in humans, particularly by
oral administration.
Summary of the Invention
In one aspect of the present invention there is provided a liquid composition
comprising a salicylate compound, a glycerol derivative, and a saccharin compound.
The composition may further comprise a flavouring agent, which may comprise or
consist of mint oil.
The composition provides a stable, liquid form of fully solubilised salicylate compound.
As a result, the Gl irritant side-effects discussed above are dramatically reduced,
avoided, ameliorated or eliminated. Furthermore the liquid form provides a very easily
administered composition. The components of the composition are pharmaceutically
acceptable and provide a composition safe for use in the therapeutic treatment of
humans.
The salicylate compound may be selected from the group consisting of: aspirin,
triflusal, diflunisal, salsalate and salicylic acid. In preferred embodiments the salicylate
compound is aspirin.
The glycerol derivative may be a glycerol derivative as described herein. In preferred
embodiments the glycerol derivative is glycerin triacetate.
The saccharin compound may be a saccharin compound as described herein. In
preferred embodiments the saccharin compound is saccharin.
In some embodiments the composition comprises or consists of a salicylate
compound, glycerin triacetate, and saccharin. In some preferred embodiments the
composition comprises or consists of aspirin, glycerin triacetate, and saccharin. In
some embodiments the composition may comprise or consist of aspirin, glycerin
triacetate, saccharin and a flavouring agent.
In some embodiments the concentration of the salicylate compound, preferably aspirin,
may be 0.5 to 3 wt% and/or the concentration of the glycerol derivative, preferably
glycerin triacetate, may be 94 to 99 wt%. In some embodiments the concentration of
saccharin compound, preferably saccharin, may be 0.1 to 3 wt%.
For the avoidance of doubt, the components are present in the composition such that
the total amount is equal to 100 wt%. For example, for a composition consisting of
salicylate compound, glycerol derivative and saccharin compound, there may be 0.5 to
3 wt% salicylate compound, preferably aspirin, and 0.1 to 3 wt% saccharin compound,
preferably saccharin. The glycerol derivative will then make up the balance of the
composition to 00 wt%.
The glycerol derivative, e.g. glycerin triacetate, may be obtained or obtainable by a
purification process. This process may be suitable to remove water or other impurities
from the glycerol derivative. For example, the glycerol derivative may be treated by
distillation and/or by passing through activated earth.
The glycerol derivative, e.g. glycerin triacetate, may be obtained or be obtainable by
passing through activated earth.
In some embodiments the composition may have a salicylate compound, e.g. aspirin,
degradation rate at 25°C of less than 0.04%/day and/or less than 0.006 mg/g/day. In
some embodiments the composition may have a salicylate compound, e.g. aspirin,
degradation rate at 25°C of less than 0.02%/day and/or less than 0.004 mg/g/day.
The composition may be free of particulates. The salicylate compound may be
completely soluble in the liquid composition. In some embodiments, greater than 90 %
of the total amount of salicylate compound, preferably aspirin, in the composition is
fully dissolved, for example at least 95 %, at least 96 %, at least 97 %, at least 98 %,
at least 99 % or at least 99.9 %. In some embodiments 100 % of the total salicylate
compound is fully dissolved.
The composition may be suitable for oral use, or formulated for intravenous or intraarterial
administration or formulated for inhalation or insufflation administration. The
composition may be a pharmaceutical composition or medicament.
!n another asoect of the es t invention a m p thn nf n n rin or n rod in a liauid
composition is provided, the method comprising admixing a salicylate compound,
glycerol derivative, and a saccharin compound. The method may further comprise
admixing a flavouring agent, e.g. mint oil.
The method may comprise comprising the steps of:
(a) adding the salicylate compound and saccharin compound to the glycerin
derivative;
(b) optionally adding a flavouring agent; and
(b) mixing until fully dissolved.
The mixing may be performed by sonication of the liquid composition.
The salicylate compound may be selected from the group consisting of: aspirin,
triflusal, diflunisal, salsalate and salicylic acid. In preferred embodiments the salicylate
compound is aspirin.
The glycerol derivative may be a glycerol derivative as described herein. In preferred
embodiments the glycerol derivative is glycerin triacetate.
The saccharin compound may be a saccharin compound as described herein. In
preferred embodiments the saccharin compound is saccharin.
In some embodiments the method comprises admixing aspirin, glycerin triacetate and
saccharin.
In some embodiments the concentration of the salicylate compound may be 0.5 to 3
wt% and/or the concentration of the glycerol derivative, preferably glycerin triacetate,
may be 94 to 99 wt%.
In some embodiments the concentration of saccharin compound, preferably saccharin,
is 0.1 to 3 wt%.
In another aspect of the present invention a packaged article comprising the liquid
composition of the present invention is sealed therein is provided. The packaged
article may be a bottle, pipette, syringe, vial, sachet, stick shot and liquid gel capsule.
The Blow-Fill-Seal (BFS) method may be used to manufacture the packaged article.
In another aspect of the present invention a method of taking aspirin which comprises
orally administering a stable liquid composition according to the present invention is
provided.
In another aspect of the present invention a method of taking aspirin which comprises
oral, rectal, nasogastric, parenteral (e.g. intravenous or intrarterial), inhalation or
insufflation administration of the liquid composition according to the present invention
is provided.
In another aspect of the present invention a liquid composition according to the present
invention is provided for use in a method of medical treatment.
In another aspect of the present invention the use of a salicylate compound or glycerol
derivative or saccharin compound in the manufacture of a medicament or
pharmaceutical composition according to the present invention for use in a method of
medical treatment is provided.
In another aspect of the present invention a method of treatment of a disease or
disorder is provided, the method comprising administering to a subject in need of
treatment the composition according to the present invention, thereby treating the
subject.
In some embodiments the method of medical treatment involves oral, rectal,
nasogastric, parenteral (e.g. intravenous or intrarterial), inhalation or insufflation
administration of the liquid composition.
In some embodiments the method of medical treatment involves the treatment or
prevention of a cardiovascular disease, cerebrovascular disease or cancer.
The cardiovascular disease may be selected from angina pectoris; heart failure (HF);
left or right ventricular failure; pulmonary heart disease; ischaemic heart disease (IHD);
cardiomyopathy; cardiac dysrhythmia; stenosis of a heart valve; hypertrophic
cardiomyopathy (HCM); coronary heart disease; paediatric cardiovascular disease
(e.g. Kawasaki disease); and congenital heart disease.
The cerebrovascular disease may be selected from thrombotic and embolic stroke,
cerebral ischaemia, brain ischaemia, transient ischaemic attack (TIA), and vascular
dementia.
The cancer may be a cancer occurring in the central nervous system, a brain cancer or
glioma.
The cancer may be a cancer of the gastrointestinal tract.
We have now surprisingly discovered an aspirin composition which overcomes or
significantly reduces at least one of the aforementioned problems.
Accordingly, in one aspect the present invention provides a stable liquid composition
comprising aspirin, glycerine triacetate, saccharin and an optional flavouring agent.
In another aspect the invention further provides a packaged article comprising a stable
liquid composition comprising aspirin, glycerine triacetate, saccharin and an optional
flavouring agent, which is sealed therein
In another aspect the invention yet further provides a method of taking aspirin which
comprises orally administering a stable liquid composition comprising aspirin, glycerine
triacetate, saccharin and an optional flavouring agent.
In another aspect the invention still further provides a stable liquid composition
comprising aspirin, glycerine triacetate, saccharin and an optional flavouring agent for
use by oral administration as an analgesic, antipyretic, anti-inflammatory and/or
antiplatelet.
In another aspect the invention even further provides the use of a composition
comprising or consisting of glycerine triacetate, saccharin and an optional flavouring
agent to produce a stable aspirin solution.
The following numbered paragraphs contain statements of broad combinations of the
inventive technical features herein disclosed:-
. A stable liquid composition comprising aspirin, glycerine triacetate, saccharin
and an optional flavouring agent.
2. The composition according to paragraph 1 wherein the concentration of the
aspirin is 0.5 to 3% and/or the concentration of the glycerin triacetate is 94 to 99%.
3. The composition according to either one of paragraphs 1 and 2 wherein the
concentration of the saccharin is 0.1 to 3%.
4 . The composition according to any one of the preceding paragraphs wherein the
flavouring agent comprises or consists of mint oil.
5. The composition according to any one of the preceding paragraphs wherein the
composition consists of aspirin, glycerine triacetate, saccharin and an optional
flavouring agent.
6 . The composition according to any one of the preceding paragraphs wherein the
glycerine triacetate is obtainable by passing through activated earth.
7. The composition according to any one of the preceding paragraphs having an
aspirin degradation rate at 25°C of less than 0.04%/day and/or less than 0.006
mg/g/day.
8. The composition according to paragraph 7 having an aspirin degradation rate
at 25°C of less than 0.02%/day and/or less than 0.004 mg/g/day.
9. The composition according to any one of the preceding paragraphs which is
free of particulates and/or the aspirin is completely soluble.
10. The composition according to any one of the preceding paragraphs which is
suitable for oral use.
11. A packaged article comprising the composition as defined in any one of the
preceding paragraphs which is sealed therein.
2 . The packaged article according to paragraph 1 selected from the group
consisting of a bottle, pipette, syringe, vial, sachet, stick shot and liquid gel capsule.
13. A method of taking aspirin which comprises orally administering a stable liquid
composition comprising aspirin, glycerine triacetate, saccharin and a optional
flavouring agent.
14. A stable liquid composition comprising aspirin, glycerine triacetate, saccharin
and an optional flavouring agent for use by oral administration as an analgesic,
antipyretic, anti-inflammatory and/or antiplatelet.
15. The use of a composition comprising or consisting of glycerine triacetate,
saccharin and an optional flavouring agent to produce a stable aspirin solution.
Description
The inventors have discovered that by formulating a salicylate compound together with
a glycerol derivative and a saccharin compound that a highly stable liquid salicylate
formulation can be provided.
Salicylate compound
The term "salicylate compound" as used herein refers to compounds according to
Formula (I):
R1 is a negative charge, or is independently selected from the group consisting of
-H,
-RA
-COH, -COR , -COOH, -COOR ,
-IMH2, -NHR , -NR 2 and -NR 2;
is independently selected from the group consisting of
-H,
-R ,
-COH, -COR , -COOH, -COOR ,
-NH2, -NHR , -NR 2, -NR 2, and
-Q;
-R6 are each independently selected from the group consisting of
-H,
-F, -CI, -Br, -I,
-RA,
-COH, -COR , -COOH, -COOR ,
-NH2, -NHR , -NR 2, -NR 2,
-SO3H, -S(0)R and -S(0 )R ;
Q is
and P is independently selected from -H, linear or branched Ci-4alkyl, alkenyl or
alkynyl, or -COR ;
wherein -R is independently selected from the group consisting of
linear or branched alkenyl or alkynyl,
phenyl optionally substituted with one or more groups -RD,
benzyl optionally substituted with one or more groups -R ,
-COOH, -COOR , -C(0)R c , -NH2, -NHR C or -NRC
2;
-RD is independently selected from
linear or branched C -4alkyl, alkenyl or alkynyl,
-F, -CI, -Br, -I,
-CN, -NO2,
-COH, -COR c , -COOH, -COOR c ,
-NH2 -NHR , -NR 2, -NR 2,
-S(0)R and -S(0 2)Rc ;
-Rc is independently selected from linear or branched C -4alkyl, alkenyl or alkynyl;
-NRB2 is independently selected from the group consisting of azetidino, imidazolidino,
pyrazolidino, pyrrolidino, piperidino, piperazino, N-Ci -4alkyl-piperazino, morpholino,
azepino or diazepino, optionally substituted with one or more groups selected from
linear or branched alkenyl or alkynyl, phenyl and benzyl; and
[C+] is an optional counter-cation selected from the group consisting of alkali metal
ions, alkaline earth metal ions, transition metal ions, Al +, ammonium or substituted
ammonium ion and N0 2
+.
The group -R1
In some embodiments, R is a negative charge, or is independently selected from the
group consisting of
-H,
-R
-COH, -COR , -COOH, -COOR ,
-NH2, -NHR , -NR 2and -NRB .
In some embodiments, R is a negative charge, or is independently selected from the
group consisting of -H and -R .
In some embodiments, R is a negative charge, or is independently selected from the
group consisting of
-H,
linear or branched C -4aik l, alkenyl or alkynyl,
phenyl optionally substituted with one or more groups -R ,
benzyl optionally substituted with one or more groups -RD,
and -C(0)R c .
In some embodiments, R1 is a negative charge, -H, or linear or branched Ci-4alkyl,
alkenyl or alkynyl.
In some embodiments, R is a negative charge.
In some embodiments, R1 is -H.
When R is a negative charge, counter-cation [C +] is present.
The group -R2
In some embodiments, R2 is independently selected from the group consisting of
-H,
-R ,
-COH, -COR , -COOH, -COOR , and
-Q.
In some embodiments, R2 is independently selected from the group consisting of -H, -
COH, -CORA and -Q.
In some embodiments, R2 is independently selected from the group consisting of -H,
-CORc , and -Q.
In some embodiments, R2 is independently selected from the group consisting of -H,
-C(0)CH 3, and -Q.
In some embodiments, R2 is independently -H.
In some embodiments, R2 is independently -C(0)CH3.
In some embodiments, R2 is independently -Q.
The groups - ?6
In some embodiments, R3-R6 are each independently selected from
-H,
-F, -CI, -Br,
-RA,
-OH, -ORA, -CF3 and -OCF3.
In some embodiments, R -R6 are each independently selected from
-H,
-F, -CI,
-R ,
-OH, -ORA and -CF3.
In some embodiments, R -R6 are each independently selected from
-H,
phenyl optionally substituted with one or more groups -RD,
and -CF3.
In some embodiments, R4 is independently -CF3, wherein R3, R5 and R6 are preferably
each independently -H.
In some embodiments, R5 is phenyl optionally substituted with one or more groups -RD,
wherein -RD is preferably -F, and R3, R4 and R6 are preferably each independently -H.
In some embodiments, each of R3-R6 is independently -H.
In some embodiments, the salicylate compound is aspirin (2-(acetoxy)benzoic acid),
according to Formula (la):
Formula (la)
In some embodiments, the salicylate compound is triflusal (2-acetyloxy-4-
(trifluoromethyl)benzoic acid), according to Formula (lb):
Formula (lb)
In some embodiments, the salicylate compound is diflunisal (2',4'-difluoro-4-
hydroxybiphenyl-3-carboxylic acid), according to Formula (Ic):
Formula (Ic)
In some embodiments, the salicylate compound is salsalate (2-(2-
hydroxybenzoyl)oxybenzoic acid), according to Formula (Id):
Formula (Id)
In some embodiments, the salicylate compound is salicylic acid (2-Hydroxybenzoic
acid), according to Formula (le):
Formula (le)
In preferred embodiments the salicylate compound is aspirin.
Glycerol derivative
The term "glycerol derivative" as used herein refers to compounds according to
Formula (II):
Formula (II)
wherein:
R -R9 are each independently selected from the group consisting of
-H,
-R ,
-COH, -COR , -COOH, -COOR ,
-NH2, -NHRA, -NR 2, -NRB
2,
-S(0)R and -S(0 2)R ;
wherein -R is independently selected from the group consisting of
linear or branched alkyl, alkenyl or alkynyl,
phenyl optionally substituted with one or more groups -RD,
benzyl optionally substituted with one or more groups -R ,
-COOH, -COOR , -C(0)R c , -NH2, -NHR or -NR 2;
-RD is independently selected from
linear or branched alkenyl or alkynyl,
-F, -CI, -Br, -I,
-OH, -OR , -CF3 -OCF3,
-CN, -N0 2,
-COH, -CORc , -COOH, -COOR ,
-NH2, -NHRC, -NRC2, -NR 2,
-S(0)R and -S(0 2)R ;
-Rc is independently selected from linear or branched Ci-4alkyl, alkenyl or alkynyl; and
-NR 2 is independently selected from the group consisting of azetidino, imidazolidino,
pyrazolidino, pyrrolidino, piperidino, piperazino, N-C 4alkyl-piperazino, morpholino,
azepino or diazepino, optionally substituted with one or more groups selected from
linear or branched C 4alkyl, alkenyl or alkynyl, phenyl and benzyl.
The groups R7-R
In some embodiments, R -R9 are each independently selected from the group
consisting of -H and -R .
In some embodiments, R -R9 are each independently selected from the group
consisting of -H, linear or branched C -4alkyl, alkenyl or alkynyl, and -C(0)R c .
In some embodiments, R -R9 are each independently selected from the group
consisting of -H, and -C(0)R c , wherein Rc is preferably methyl.
In some embodiments, one of R -R9 is -H and the other two are -C(0)R c , wherein Rc is
preferably methyl.
In some embodiments, one of R -R9 is -C(0)R c and the other two are -H, wherein Rc is
preferably methyl.
In some embodiments, R -R9 are each selected from -C(0)R c , wherein Rc is
preferably methyl.
In some preferred embodiments, the glycerol derivative is glycerin triacetate (also
known as glyceryl triacetate, GTA, or triacetin, or 1,3-Diacetyloxypropan-2-yl acetate)
according to Formula (Ha):
Formula (Ma.)
Glycerin triacetate is an FDA approved food additive with "generally regarded as safe"
status that has been tested for parenteral nutrition in a wide variety of species with no
adverse effects. Glycerin triacetate is also used as a vaporising agent in e-cigarettes
and is regarded as safe when used in that context.
The glycerin triacetate (also commonly known as glycerine triacetate, glyceryl
triacetate, or triacetin) used herein acts as a solvent for the aspirin or salicylate
compound and can give clear aspirin or salicylate compound solutions. Food grade
glycerin triacetate may be employed, but it is preferably subjected to further purification
processes, e.g. additional distillation steps. In one preferred embodiment, the glycerin
triacetate is passed through activated earth, such as a column or fixed bed thereof.
The viscosity of glycerin triacetate can be reduced by mixing with a suitable solvent,
such as ethanol, prior to passing through the activated earth. The solvent can then be
removed by using vacuum distillation followed by steam distillation, preferably to levels
below 1 ppm of solvent in the glycerine triacetate.
Saccharin compound
The term "saccharin compound" as used herein refers to compounds according to
Formula (111):
Formula (III)
wherein:
1 -R are each independently selected from the group consisting of
-COH, -COR A , -COOH, -COOR A ,
-NH 2, -NHR , -NR A2, -NR ,
-SO3H, -S(0)R , -S(0 2)R , and
-W;
X is independently selected from the group consisting of
-NQ, -NH and -NR ,
Y is independently selected from S(C>2);
wherein -R is independently selected from the group consisting of
linear or branched Ci-4alkyl, alkenyl or alkynyl,
phenyl optionally substituted with one or more groups -R ,
benzyl optionally substituted with one or more groups -R ,
-COOH, -COOR c , -C(0)R , -NH 2, -NHR or -NR C
2;
-R is independently selected from
linear or branched alkenyl or alkynyl,
-F, -CI, -Br, -I,
-CN, -N0 2,
-COH, -COR c , -COOH, -COOR c ,
-NH 2, -NHR , -NR 2, -NR B
2,
-S(0)R c and -S(0 2)R ;
-Rc is independently selected from linear or branched Ci-4alkyl, alkenyl or alkynyl;
-NR 2 is independently selected from the group consisting of azetidino, imidazolidino,
pyrazolidino, pyrrolidino, piperidino, piperazino, N-Ci-4alkyl-piperazino, morpholino,
azepino or diazepino, optionally substituted with one or more groups selected from
linear or branched Chalky!, alkenyl or alkynyl, phenyl and benzyl;
-W is the group
wherein L is independently selected from a 5- or 6-membered heteroaromatic
group and LB is independently selected from a 5- or 6-membered monosaccharide
moiety; and
[C+] is an optional counter-cation selected from the group consisting of alkali metal
ions, alkaline earth metal ions, transition metal ions, Al +, ammonium or substituted
ammonium ion and 0 2+.
The group X
In some embodiments, X is independently selected from NQ and NH.
In some embodiments, X is independently NQ and [C +] is a counter-cation selected
from alkali metal ions and alkaline earth metal ions.
In some embodiments, X is independently NQ and [C +] is a counter-cation selected
from sodium and calcium ions.
In some embodiments, X is independently NH.
The groups R10-R13
In some embodiments, R -R13 are each independently selected from
-H,
-F, -C , -Br,
-RA,
-COH, -COR , -COOH, -COOR ,
-NH2 > -NHR -NR 2 -NR 2, and
-W.
some embodiments, R 0-R13 are each independently selected from
-H,
-F, -CI, -Br,
-R ,
-OH, -OR , -CF3, -OCF3, and
-W.
In some embodiments, R -R13 are each independently selected from
-H,
-F, -CI,
- A,
-OH, -OR , and
-W.
In some embodiments, R -R13 are each independently selected from
-H,
-F, -CI,
linear or branched Ci-4alkyl, alkenyl, alkynyl,
-OH, and
-W.
In some embodiments, R -R13 are each independently selected from
-H,
-F, -CI,
linear or branched d^alkyl, alkenyl, alkynyl and
-OH.
In some embodiments, one of R -R 3 is independently -W and the remaining groups of
R 0-R13 are independently -H.
In some embodiments, R 1 is independently -W and R 0, R 2 and R 3 are each
independently -H.
The group LA
In some embodiments, L is a 5- or 6-membered heteroaromatic group independently
selected from imidazolyl, pyrazolyl, oxazolyl, isooxazolyl, thiazolyl, isothiazolyl,
dioxolanyl, dithiolanyl, triazolyl, furanyl, oxadiazolyl, thiadiazolyl, dithiazolyl, tetrazolyl,
pyridinyl, pyranyl, thiopyranyl, diazinyi, oxazinyi, thiazinyl, dioxinyl, dithiinyl, triazinyl and
tetrazinyl.
In some embodiments, L is a 5-membered heteroaromatic group independently
selected from imidazolyl, pyrazolyl, oxazolyl, isooxazolyl, thiazolyl, isothiazolyl,
dioxolanyl, dithiolanyl, triazolyl, furanyl, oxadiazolyl, thiadiazolyl, dithiazolyl and
tetrazolyl.
In some embodiments, L is a 5-membered heteroaromatic group independently
selected from triazolyl, furanyl, oxadiazolyl, thiadiazolyl and dithiazolyl.
In some embodiments, L is a 5-membered heteroaromatic group independently
selected from one of the followin triazolyl moieties:
In some embodiments, L i:s
The group LB
In some embodiments, LB is independently selected from ribofuranyl, glucopyranyl,
galactopyranyl, mannopyranyl and allopyranyl.
In some embodiments, L is the following 6-membered monosaccharide moiety:
In some preferred embodiments, the saccharin compound is saccharin (2 - 1l6,2-
benzothiazol-1 , ,3-trione), according to Formula (Ilia):
Formula (Ilia)
In some embodiments, the saccharin compound is a saccharide salt according to
Formula (lllb):
Formula (lllb)
wherein [C +] is a counter-cation selected from the group consisting of alkali metal
ions, alkaline earth metal ions, transition metal ions, Al + , ammonium or substituted
ammonium ion and N , more preferably sodium or calcium ion.
In some embodiments, the saccharin compound is a compound according to Formula
(lllc):
Formula (lllc)
Any combination of salicylate compound, glycerol derivative and saccharin compound
may be present, along with an optional flavouring agent, in the liquid compositions
according to a first aspect of the invention.
In some embodiments, the glycerol derivative is a compound according to Formula
( s ), the saccharin compound is a compound according to Formula (Ilia) or (lllb) and
the salicylate compound is one or more salicylate compounds according to Formula (I).
In some embodiments, the glycerol derivative is a compound according to Formula
(lla), the saccharin compound is a compound according to Formula (Ilia) or (lllb) and
the salicylate compound is one or more salicylate compounds according to Formulae
(la)-(le).
In some embodiments, the glycerol derivative is a compound according to Formula
(lla), the saccharin compound is a compound according to Formula (Ilia) or (lllb) and
the salicylate compound is a compound according to Formula (la).
Isomers
Certain of the compounds described above may exist in one or more particular
geometric, optical, enantiomeric, diastereoisomeric, epimeric, atropic, stereoisomeric,
tautomeric, conformational, or anomeric forms, including but not limited to, cis- and
trans-forms; E- and Z-forms; c-, t-, and r-forms; endo- and exo-forms; R-, S-, and
eso-forms; D- and L-forms; d- and I-forms; (+) and (-) forms; keto-, eno!-, and
eno!ate forms; syn- and anti-forms; synclinal- and anticlinal-forms: a- and b-forms;
axial and equatorial forms; boat-, chair-, twist-, envelope-, and halfchair-forms; and
combinations thereof, hereinafter collectively referred to as "isomers" (or "isomeric
forms").
A reference to a class of structures may well include structurally isomeric forms falling
within that class (e.g., Ci-7alkyl includes n-propyl and iso-propyl; butyl includes n-, iso-,
sec-, and tert-butyl; methoxyphenyl includes ortho-, meta-, and para-methoxyphenyl).
However, reference to a specific group or substitution pattern is not intended to include
other structural (or constitutional isomers) which differ with respect to the connections
between atoms rather than by positions in space. For example, a reference to a
methoxy group, -OCH3, is not to be construed as a reference to its structural isomer, a
hydroxymethyl group, -CH2OH. Similarly, a reference to ortho-chlorophenyl is not to
be construed as a reference to its structural isomer, meta-chlorophenyl.
The above exclusion does not pertain to tautomeric forms, for example, keto-,
and enolate forms, as in, for example, the following tautomeric pairs: keto/enol
(illustrated below), imine/enamine, amide/imino alcohol, amidine/amidine,
nitroso/oxime, thioketone/enethiol, N nitroso/hydroxyazo, and nitro/aci-nitro.
\ OH . , c-c === c=c === c=c
I \ \ keto enol enolate
Note that specifically included in the term "isomer" are compounds with one or more
isotopic substitutions. For example, H may be in any isotopic form, including 1H, 2H
(D), and 3H (T); C may be in any isotopic form, including 12C, 3C, and 4C; O may be in
any isotopic form, including 60 and 0 ; and the like.
Unless otherwise specified, a reference to a particular compound includes all such
isomeric forms, including mixtures (e.g., racemic mixtures) thereof. Methods for the
preparation (e.g., asymmetric synthesis) and separation (e.g., fractional crystallisation
and chromatographic means) of such isomeric forms are known in the art or are
readily obtained by adapting known methods in a known manner.
Salts
As explained above, the salicylate compound and/or the saccharin compound can be
provided as salts. As such, in this specification the terms "salicylate compound" and
"saccharin compound" include salts thereof.
It may be convenient or desirable to prepare, purify, and/or handle a corresponding
salt of a compound, for example, a pharmaceutically-acceptable salt. Examples of
pharmaceutically acceptable salts are discussed in Berge et al., 1977,
"Pharmaceutically Acceptable Salts," J . Pharm. Sci., Vol. 66, pp. 1-19.
For example, when the salicylate compound or saccharin compound exists as an
anion, or has a functional group which may be anionic (e.g., -COOH may be -COO ) ,
then a salt may be formed with a suitable cation (such as [C +] described above).
Examples of suitable inorganic cations include, but are not limited to, alkali metal ions
such as Na+ and K+, alkaline earth cations such as Ca2+ and Mg2+, and other cations
such as Al + . Examples of suitable organic cations include, but are not limited to,
ammonium ion (i.e., NH +) and substituted ammonium ions (e.g., NHsR+, R2 ,
NHR +, NR +). Examples of some suitable substituted ammonium ions are those
derived from: ethylamine, diethylamine, dicyclohexylamine, triethylamine, butylamine,
ethylenediamine, ethanolamine, diethanolamine, piperazine, benzylamine,
phenylbenzylamine, choline, meglumine, and tromethamine, as well as amino acids,
such as lysine and arginine. An example of a common quaternary ammonium ion is
N(CH3) +.
If a compound is cationic, or has a functional group which may be cationic (e.g., -NH2
may be -NhV), then a salt may be formed with a suitable anion. Examples of suitable
inorganic anions include, but are not limited to, those derived from the following
inorganic acids: hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfurous, nitric,
nitrous, phosphoric, and phosphorous. Examples of suitable organic anions include,
but are not limited to, those derived from the following organic acids:
2-acetyoxybenzoic, acetic, ascorbic, aspartic, benzoic, camphorsulfonic, cinnamic,
citric, edetic, ethanedisulfonic, ethanesulfonic, fumaric, glucheptonic, gluconic,
glutamic, glycolic, hydroxymaleic, hydroxynaphthalene carboxylic, isethionic, lactic,
!actobionic, !auric, ma!eic, malic, methanesulfonic, mucin, oleic, oxalic, palmitic,
pamoic, pantothenic, phenylacetic, phenylsulfonic, propionic, pyruvic, salicylic, stearic,
succinic, sulfanilic, tartaric, toluenesulfonic, and valeric. Examples of suitable
polymeric organic anions include, but are not limited to, those derived from the
following polymeric acids: tannic acid, carboxymethyl cellulose.
Unless otherwise specified, a reference to a particular compound also includes salt
forms thereof.
Solvates and Hydrates
It may be convenient or desirable to prepare, purify, and/or handle a corresponding
solvate of a compound. The term "solvate" is used herein in the conventional sense to
refer to a complex of solute (e.g., compound, salt of compound) and solvent. If the
solvent is water, the solvate may be conveniently referred to as a hydrate, for example,
a mono-hydrate, a di hydrate, a tri-hydrate, etc.
Unless otherwise specified, a reference to a particular compound also includes solvate
and hydrate forms thereof.
Prodrugs
It may be convenient or desirable to prepare, purify, and/or handle a compound,
especially the salicylate compound, in the form of a prodrug. The term "prodrug," as
used herein, pertains to a compound which, when metabolised (e.g., in vivo), yields the
desired active compound. Typically, the prodrug is inactive, or less active than the
desired active compound, but may provide advantageous handling, administration, or
metabolic properties.
In one embodiment, the glycerin triacetate used herein may suitably comprise in the
range from 0 to 1 wt%, preferably 0 to 0.5 wt%, more preferably 0 to 0.1 wt%,
particularly 0 to 0.05 wt%, and especially 0 to 0.015% of glycerin monoacetate and/or
glycerine diacetate.
The glycerin triacetate may also suitably comprise in the range from 0 to 100 ppm,
preferably 0 to 20 ppm, more preferably 0 to 10 ppm, particularly 0 to 2 ppm, and
especially 0 to 1 ppm of glycerine monoacetate and/or glycerine diacetate.
In one embodiment, the glycerin triacetate may suitably comprise in the range from 0
to 50 ppm, preferably 0 to 10 ppm, more preferably 0 to 5 ppm, particularly 0 to 1 ppm,
and especially 0 to 0.5 ppm of glycerine.
The glycerin triacetate suitably comprises in the range from 0 to 100 ppm, preferably 0
to 20 ppm, more preferably 0 to 10 ppm, particularly 0 to 2 ppm, and particularly 0 to 1
ppm of non-polar materials, such as colour pigments and/or soaps.
In one preferred embodiment, the glycerin triacetate is substantially anhydrous,
suitably comprising in the range from 0 to 0.5 wt%, preferably less than 0.3 wt%, more
preferably less than 0.2 wt%, particularly less than 0.1 wt%, and especially less than
0.05 wt% of water.
A reduction in water content of the glycerin triacetate ensures a low water content in
the composition. This provides increased stability of the liquid composition because
salicylate compounds such as aspirin are vulnerable to hydrolysis in the presence of
water to produce salicylic acid and other acids as by-products. If an aspirin
composition contains 10 wt% salicylic acid with reference to the total amount of aspirin
and salicylic acid present, the composition is no longer deemed to be pharmaceutically
acceptable. Thus an increase in aspirin stability results in an extended shelf-life of the
pharmaceutical composition.
Preferably the composition comprises at least 5 wt% glycerin triacetate, for example at
least 10 wt%, at least 15 wt%, at least 20 wt% or at least 25 wt%. Most preferably, the
composition comprises at least 90 wt% glycerin triacetate, for example at least 9 1 wt%,
at least 92 wt%, at least 93 wt%, at least 94 wt%, at least 95 wt% or at least 96 wt%.
Preferably the composition comprises up to 99.9 wt% glycerin triacetate, for example
up to 99 wt%, up to 98 wt%, up to 97 wt% or up to 96.5 wt%.
The concentration of the glycerin triacetate in the composition according to the
invention is suitably in the range from 90 to 99%, preferably 94 to 98%, more
preferably 95 to 97.5%, particularly 95.5 to 97%, and especially 96 to 96.5% by weight
s d n tj-iQ total weight of the composition.
At this concentration of glycerin triacetate excellent solubilisation and stability of the
salicylate compound is observed and a shelf-life of over 1 year, preferably about 18
months or 2 years, may be achieved. As used herein, "stability" refers to the
resistance to degradation of the salicylate compound. Higher stability means that after
a given period of time lower levels of contaminants will accumulate in the composition.
These contaminants may be the products of the hydrolysis of the salicylate compound.
The stability of a composition may be measured using the method described below.
Preferably the composition comprises at least 0.1 wt% salicylate compound, e.g.
aspirin, for example at least 0.5 wt%, at least 1 wt%, at least 1.5 wt%, at least 2.0 wt%
or at least 2.5 wt%. In this way a pharmaceutically effective amount of salicylate
compound, e.g. aspirin may be provided in a small amount of liquid composition. Of
course the necessary dose will depend on e.g. the disorder being treated, the identity
of the patient etc. as explained further below.
Preferably the composition comprises up to 10 wt% salicylate compound, e.g. aspirin,
for example up to 9 wt%, up to 8 wt%, up to 7 wt%, up to 6 wt%, up to 5 wt%, up to 4
wt%, up to 3 wt% or up to 2.5 wt%. In this way the stability and solubility of the
salicylate compound, e.g. aspirin, in the composition is maximised.
When the concentration of salicylate compound, e.g. aspirin, exceeds this amount
there is an increased risk that not all the aspirin in the liquid composition will be fully
solubilised, which may lead to a granular precipitate having the disadvantages
associated with solid aspirin compositions.
The concentration of the salicylate compound, e.g. aspirin, in the composition
according to the invention is suitably in the range from 0.5 to 7%, preferably 1 to 5%,
more preferably 2 to 3%, particularly 2.3 to 2.7%, and especially 2.4 to 2.6% by weight
based on the total weight of the composition.
In one preferred embodiment, the salicylate compound is substantially anhydrous,
suitably comprising in the range from 0 to 0.5 wt%, preferably less than 0.3 wt%, more
preferably less than 0.2 wt%, particularly less than 0.1 wt%, and especially less than
0.05 wt% of water.
The composition may comprise at least 0.1 wt% saccharin, for example at least 0.2
wt%, at least 0.3 wt%, at least 0.4 wt%, at least 0.5 wt%, at least 0.6 wt%, at least 0.7
wt%, at least 0.8 wt% or at least 0.9 wt%. In this way, excellent stability and
solubilisation of the salicylate compound is achieved.
The composition may comprise up to 5 wt% saccharin, for example up to 4 wt%, up to
3 wt%, up to 2 wt%, up to 1.5 wt%, up to 1.2 wt% or up to 1. 1 wt%.
The concentration of the saccharin in the composition according to the invention is
suitably in the range from 0.1 to 4%, preferably 0.4 to 3%, more preferably 0.7 to
1.5%, particularly 0.8 to 1.2%, and especially 0.9 to 1.1% by weight based on the total
weight of the composition.
In one preferred embodiment, the saccharin compound is substantially anhydrous,
suitably comprising in the range from 0 to 0.5 wt%, preferably less than 0.3 wt%, more
preferably less than 0.2 wt%, particularly less than 0.1 wt%, and especially less than
0.05 wt% of water.
The composition according to the invention is preferably substantially anhydrous,
suitably comprising in the range from 0 to 0.5 wt%, preferably less than 0.3 wt%, more
preferably less than 0.2 wt%, particularly less than 0.1 wt%, and especially less than
0.05 wt% of water.
The composition according to the invention preferably comprises less than 0.5 wt%
water, for example less than 0.4 wt%, less than 0.3 wt%, less than 0.2 wt%, or less
than 0.1 wt% water. In some embodiments the composition according to the invention
comprises less than 000 ppm water, for example less than 900 ppm, less than 800
ppm, less than 700 ppm, less than 600 ppm or less than 500 ppm water.
As explained above, keeping the level of water in the composition low increases the
stability of the salicylate compound in the composition thereby increasing the shelf-life.
The liquid composition of the invention may optionally also comprise one or more other
pharmaceutically acceptable Ingredients well known to those skilled in the art,
including, but not limited to, pharmaceutically acceptable carriers, diluents, excipients,
adjuvants, fillers, buffers, preservatives, anti-oxidants, lubricants, stabilisers,
solubilisers, surfactants (e.g., wetting agents), masking agents, colouring agents,
flavouring agents, and sweetening agents. The formulation may further comprise
other active agents, for example, other therapeutic or prophylactic agents.
The term "pharmaceutically acceptable," as used herein, pertains to compounds,
ingredients, materials, compositions, dosage forms, etc., which are, within the scope of
sound medical judgment, suitable for use in contact with the tissues of the subject in
question (e.g., human) without excessive toxicity, irritation, allergic response, or other
problem or complication, commensurate with a reasonable benefit/risk ratio. Each
carrier, diluent, excipient, etc. must also be "acceptable" in the sense of being
compatible with the other ingredients of the formulation without exerting a detrimental
effect on the solubility or stability of the salicylate compound.
Suitable carriers, diluents, excipients, etc. can be found in standard pharmaceutical
texts, for example, Remington's Pharmaceutical Sciences , 18th edition, Mack
Publishing Company, Easton, Pa., 1990; and Handbook of Pharmaceutical Excipients ,
5th edition, 2005.
The composition may also comprise an optional masking or flavouring agent. Suitable
masking agents may mask or hide any objectionable flavour of the base composition.
The flavouring agent preferably comprises any natural or synthetically prepared fruit or
botanical flavouring agent, or mixtures thereof. Suitable natural or artificial fruit
flavouring agents include lemon, orange, grapefruit, strawberry, banana, pear, kiwi,
grape, apple, mango, pineapple, passion fruit, raspberry, and mixtures thereof. The
fruit flavouring agent is suitably in anhydrous form such as dried juice or oil, preferably
oil. Suitable botanical flavours include any member of the mint family such as
spearmint, peppermint, watermint, apple mint; and other flavours such as Jamaica,
marigold, chrysanthemum, tea, chamomile, ginger, valerian, yohimbe, hops,
eriodictyon, ginseng, bilberry, rice, red wine, mango, peony, lemon balm, nut gall, oak
chip, lavender, walnut, gentiam, luo han guo, cinnamon, angelica, aloe, agrimony,
yarrow, and mixtures thereof. The botanical flavouring agent is suitably an anhydrous
concentrate or an extract, for example in the form of oil or dried to form a powder.
The flavouring agent preferably comprises, consists essentially of, or consists of, mint
flavouring agent, more preferably in the form of oil, particularly spearmint and/or
peppermint oil, and especially spearmint oil.
The composition may comprise at least 0 wt%, for example at least 0.05 wt%, at least
0.06 wt%, at least 0.07 wt%, at least 0.08 wt%, at least 0.09 wt% or at least 0.1 wt%
flavouring agent, preferably oil, more preferably mint oil.
In this way, an additional stability enhancement of the salicylate compound may be
observed.
The composition may comprise up to 3 wt%, for example up to 2.5 wt%, up to 2 wt%,
up to 1.5 wt%, up to 1 wt%, up to 0.5 wt%, up to 0.2 w†%, up to 0.18 wt% or up to 0.16
wt% flavouring agent, preferably oil, more preferably mint oil.
The concentration of flavouring agent, preferably oil, in the composition is suitably in
the range from 0 to 3%, preferably 0.05 to 1%, more preferably 0.1 to 0.2%,
particularly 0.12 to 0.18%, and especially 0.14 to 0.16% by weight based on the total
weight of the composition.
The composition may also comprise an optional, additional natural or artificial
sweetener or sweetening agent (in addition to saccharin). Suitable sweeteners are
natural sugars which may be granulated or powdered, and include sucrose, fructose,
dextrose, maltose, lactose, xylitol, polyols, and mixtures thereof.
In other embodiments, artificial sweeteners may be utilized in the composition.
Suitable optional artificial sweeteners (in addition to saccharin) include, for example,
cyclamates, sucralose, acesulfam-K, L-aspartyl-L-phenylalanine lower alkyl ester
sweeteners (e.g. aspartame), L-aspartyl 1-D -alanine amides, L-aspartyl-D-serine
amides, L-aspartyl-L-1-hydroxymethylalkaneamides, L-aspartyl-1-
hydroxyethyalkaneamides, L-aspartyl-D-phenylglycine esters and amides.
The concentration of the optional, preferably artificial, sweetener (not including
saccharin) in the composition is suitably in the range from 0 to 5%, preferably 0.4 to
2%, more preferably 0.7 to 1.3%, particularly 0.8 to 1.2%, and especially 0.9 to 1.1%
by weight based on the total weight of the composition.
The composition according to the present invention may also contain an antioxidant.
Suitable examples of antioxidants include a phenolic compound, a plant extract, or a
sulphur-containing compound. The antioxidant may be ascorbic acid or a salt thereof,
vitamin E, CoQIO, tocopherols, lipid soluble derivatives of more polar antioxidants such
as ascorbyl fatty acid esters (e.g. ascorbyl palmitate), plant extracts (e.g. rosemary,
sage and oregano oils, green tea extract), algal extracts, and synthetic antioxidants
(e.g., butylated hydroxytoluene (BHT), tert-butylhydroquinone (TBHQ), butylated
hydroxyanisole (BHA), ethoxyquin, alkyl gallates, hydroquinones, tocotrienols), and
combinations thereof.
The concentration of the antioxidant in the composition is preferably in the range from
0 to 2%, more preferably 0.01 to 1.5%, particularly 0.1 to 1%, and especially 0.15 to
0.5% by weight based on the total weight of the composition.
The composition according to one embodiment of the invention comprises, consists
essentially of, or consists of, aspirin, glycerin triacetate, saccharin and an optional
flavouring agent.
The aspirin is preferably completely soluble in the composition and the composition is
more preferably free of particulate material, as described above. The composition
exhibits surprisingly improved stability to degradation. We have discovered that the
flavouring agent, particularly mint oil, can also surprisingly contribute to the improved
stability to degradation.
The composition according to the present invention is preferably stable, measured as
described herein, suitably having a salicylate compound, e.g. aspirin degradation rate
at 25°C of less than 0.05%/day, preferably less than 0.04%/day, more preferably less
than 0.03%/day, particularly less than 0.02%/day, and especially less than
0.015%/day.
The composition suitably has a salicylate compounds, e.g. aspirin degradation rate in
mg per gram of composition per day at 25°C, measured as described herein, of less
than 0.01 mg/g/day, preferably less than 0.007 mg/g/day, more preferably less than
0.006 mg/g/day, particularly less than 0.005 mg/g//day, and especially less than 0.004
mg/g/day.
The liquid composition described herein is particularly suitable for oral use as an
alternative to normal aspirin tablets. One particular use is to orally administer to
stroke or heart attack victims immediately after the attack, e.g. suitably within 12 hours,
preferably within 8 hours, more preferably within 4 hours, particularly within 2 hours,
and especially within 1 hour of the heart attack, e.g. in the ambulance on the way to
hospital.
Formulations
Liquid compositions according to the present invention are preferably formulated as a
pharmaceutical composition or medicament.
Thus, in some embodiment the liquid composition is a pharmaceutical composition or
medicament. Methods of making a pharmaceutical composition or medicament
comprising admixing the salicylate compound, glycerol derivative and saccharin
compound are provided, which methods may optionally further comprise admixing
together with one or more other pharmaceutically acceptable ingredients well known to
those skilled in the art, e.g., carriers, adjuvants, excipients, etc. If formulated as
discrete units (e.g., tablets / capsules, etc.), each unit contains a predetermined
amount (dosage) of the active compound(s), i.e. the salicylate compound and/or
glycerol derivative and/or saccharin compound.
The term "pharmaceutically acceptable" as used herein pertains to compounds,
ingredients, materials, compositions, dosage forms, etc., which are, within the scope of
sound medical judgment, suitable for use in contact with the tissues of the subject in
question (e.g., human) without excessive toxicity, irritation, allergic response, or other
problem or complication, commensurate with a reasonable benefit/risk ratio. Each
carrier, adjuvant, excipient, etc. must also be "acceptable" in the sense of being
compatible with the other ingredients of the formulation.
Suitable carriers, adjuvants, excipients, etc. can be found in standard pharmaceutical
texts, for example, Remington's Pharmaceutical Sciences , 18th edition, Mack
Publishing Company, Easton, Pa., 1990; and Handbook of Pharmaceutical Excipients ,
2nd edition, 1994.
Pharmaceutical formulations may be prepared by any methods well known in the art of
pharmacy. Such methods include the step of bringing into association the active
compound with a carrier which constitutes one or more accessory ingredients. In
general, the formulations are prepared by uniformly and intimately bringing into
association the active compound with carriers (e.g., liquid carriers, finely divided solid
carrier, etc.), and then shaping the product, if necessary.
The formulation may be prepared to provide for rapid or slow release; immediate,
delayed, timed, or sustained release; or a combination thereof.
Formulations may suitably be in the form of liquids, elixirs, syrups, mouthwashes,
drops, capsules (including, e.g., gel capsules, hard and soft gelatin capsules),
ampoules, sprays, mists, aerosols or vapours. Liquid formulations may be suitable for
oral administration, e.g. in the form of a gel capsule filled with the liquid composition, or
for intravenous or intraarterial administration, e.g. by injection. Liquid formulations may
also be suitable for inhalation through the nose or mouth, e.g. when delivered as an
aerosol or vapour.
Formulations suitable for inhalation administration, include, for example, nasal spray,
nasal drops, or by aerosol administration by nebuliser. Formulations suitable for
pulmonary administration (e.g., by inhalation or insufflation therapy) include those
presented as an aerosol spray from a pressurised pack, with the use of a suitable
propellant, such as dichlorodifluoromethane, trichlorofluoromethane, dichorotetrafluoroethane,
carbon dioxide, or other suitable gases.
Formulations may suitably be provided as a patch, adhesive plaster, bandage,
dressing, or the like which is impregnated with the liquid composition and optionally
one or more other pharmaceutically acceptable ingredients, including, for example,
penetration, permeation, and absorption enhancers.
Methods of Treatment
The liquid compositions described herein are useful in the treatment or prevention of
diseases or disorders including, for example, those which are known to be treated with,
or known to be treatable with salicylate compounds (e.g. aspirin).
In some aspects of the present invention the liquid composition is provided for use in a
method of treatment of the human or animal body by therapy, for example, for use in a
method of treatment of a disease or disorder.
Preferably, the beneficial therapeutic effects of the treatment outweigh any potentially
harmful effects of the salicylate compound. For example, some salicylate compounds
were thought to be associated with Reye's syndrome in children. In reality Reye's
syndrome has only been observed rarely in children with Kawasaki Disease treated
with high dose aspirin (Buck ML., 'Use of Aspirin in children with cardiac disease',
Pediatr Pharm 2007;13(1), incorporated herein by reference). There is in fact no
convincing evidence linking the ingestion of Aspirin in children with the cause of Reye's
syndrome. Most authorities now attribute Reye's syndrome to a viral aetiology. The
implication of aspirin as a causative agent was due to the confounding factor that most
of these children had a fever and were administered aspirin. Of course, the trained
medical practitioner can assess such risk balanced against the benefits of the
particular treatment. All children with Kawasaki disease are currently prescribed
aspirin (often in high dose) as the risk: enefit ratio vastly favours benefit over risk.
In some aspects of the present invention a method of treatment is provided for the
prevention or treatment of a disease or disorder, the method comprising administering
to a patient n need of treatment a therapeutically effective amount of a liquid
composition according to the present invention.
Another aspect of the present invention pertains to a method of treatment, for
example, of a disorder (e.g., a disease) as described herein, comprising administering
to a patient in need of treatment a therapeutically effective amount of a liquid
composition, as described herein, preferably in the form of a pharmaceutical
composition, and one or more (e.g., 1, 2, 3, 4) additional therapeutic agents, as
described herein, preferably in the form of a pharmaceutical composition. In some
embodiments the additional therapeutic agent may be administered in a separate
composition, such administration being simultaneous, separate or sequential to the
liquid composition of the present invention. The liquid composition of the present
invention and additional therapeutic agent may be presented together in the form of a
kit, optionally with instructions for their use. In other embodiments the additional
therapeutic agent may be formulated together with the liquid composition of the
present invention. In some embodiments where the disease being treated is a cancer
an additional therapeutic agent may be a chemotherapeutic agent, e.g. temozolomide.
The term "treatment," as used herein in the context of treating a condition, pertains
generally to treatment and therapy, whether of a human or an animal (e.g., in
veterinary applications), in which some desired therapeutic effect is achieved, for
example, the inhibition of the progress of the condition, and includes a reduction in the
rate of progress, a halt in the rate of progress, alleviation of symptoms of the condition,
amelioration of the condition, and cure of the condition. Treatment as a prophylactic
measure (i.e., prophylaxis) is also included. For example, use with patients who have
not yet developed the condition, but who are at risk of developing the condition, is
encompassed by the term "treatment."
For example, treatment of glioblastoma multiforme includes the prophylaxis of
glioblastoma multiforme, reducing the incidence of glioblastoma multiforme, reducing
the severity of glioblastoma multiforme, alleviating the symptoms of glioblastoma
multiforme, etc.
The term "therapeutically-effective amount," as used herein, pertains to that amount of
a compound, or a material, composition or dosage form comprising a compound,
which is effective for producing some desired therapeutic effect, commensurate with a
reasonable benefit/risk ratio, when administered in accordance with a desired
treatment regimen.
Diseases or disorders to be treated or prevented may include any disease or disorder
for which a salicylate compound, glycerol derivative or saccharin compound has a
treatment effect, which may be a protective, preventative or prophylactic effect.
Diseases or disorders to be treated may include fever, pain, inflammation, or swelling.
Methods of treatment may also involve prevention of formation of blood clots, bloodthinning,
or reduction of clot formation, prevention or reduction of heart attack, stroke
or ischaemia, prevention or treatment of cancer.
In some embodiments a disease or disorder to be treated or prevented may be a
cardiovascular disease, cerebrovascular disease or a cancer.
Salicylate compounds are known for the treatment or prevention of a range of
cardiovascular and cerebrovascular disorders (e.g. see Hall, SL; Lorenc, T ( 1 February
2010). "Secondary prevention of coronary artery disease". American family physician
8 1 (3): 289-96, incorporated herein by reference), for use in the treatment of
prevention of cancer (e.g. see Fengming Lan et al., Antitumor effect of aspirin in
glioblastoma cells by modulation of b-catenin/T-cell factor-mediated transcriptional
activity. J Neurosurg 115:780-788, 201 1, incorporated herein by reference), and as
non-steroidal anti-inflammatory (NSAID) agents (e.g. see Morris et al., Effects of Low-
Dose Aspirin on Acute Inflammatory Responses in Humans. The Journal of
Immunology. 2009; 183:2089-2096, incorporated herein by reference).
Aspirin is a well-known salicylate drug widely used as an analgesic, antipyretic, anti
inflammatory and anti-platelet and therefore finds application in the treatment or
prevention of pain, fever, and inflammation. Its anticoagulant action has made it
popular in the treatment or prevention of cardiovascular diseases such as heart attack,
stroke, and ischaemia.
The anti-cancer effect of aspirin was first reported in 1972 (Gasic Gl, et al. Lancet.
1972;2:932-937) and is well known. Recently, Fengming Lan et al; (supra) describe
aspirin to be a potent antitumor agent, capable of inhibiting glioma cell proliferation and
invasive ability and inducing apoptotic cell death. Low dose, long term aspirin use is
proposed as having a future role in preventing glioma or as an adjuvant therapeutic
agent in the treatment of glioma (Matthew W. Brown., Characterisation of the effects of
chronic aspirin treatment on the viability and proliferation of stage 4 glioblastoma cells.
Diffusion: the UCLan Journal of Undergraduate Research Volume 6 Issue 2 December
2013, incorporated herein by reference).
See also the additional references cited in the background section, above.
Glycerin triacetate freely crosses the blood-brain barrier and has been shown to be
effective as an adjuvant in the chemotherapeutic treatment of glioma slowing the
growth of glioma stem cells in culture but not affecting normal glial cells or neural stem
cells (e.g. see Tsen et al., Triacetin-based acetate supplementation as a
chemotherapeutic adjuvant therapy in glioma. Int J Cancer. 2014 March 15;
134(6): 1300-1 310; and Long et a/., 'Acetate Supplementation Induces Growth Arrest of
NG2/PDGFRa-Positive Oligodendroglioma-Derived Tumor-Initiating Cells', PLoS One,
2013, e80714, both incorporated herein by reference). Glycerin triacetate enhanced
the efficacy of temozolomide against glioma when the two agents were administered in
combination. The inventors expected glycerin triacetate to provide a similar effect in
salicylate compound, e.g. aspirin, therapy, to enhance the delivery of the salicylate
compound across the blood-brain barrier when the two are administered in
combination, as in the present liquid compositions. A synergistic effect may thus be
observed for the salicylate compound treatment of cerebrovascular disorders such as
stroke or dementia with the present compositions. A synergistic effect may also be
observed for the combination of aspirin, saccharin and glycerin triacetate.
Furthermore, since glycerin triacetate transports aspirin across the blood-brain barrier,
the efficacy of the treatment will increase.
Given that both glycerin triacetate and aspirin have been shown to have
chemotherapeutic properties, the liquid compositions of the invention containing these
compounds provide a highly efficacious combination therapy against cancer, in
particular brain cancer, especially primary brain cancer and gliomas such as
glioblastoma multiforme. The stable and fully solubilised nature of the compositions
means that such combination therapy can be very easily provided. Additionally, since
Tsen et al. (supra) demonstrated that glycerin triacetate enhances temozolomide
chemotherapeutic efficacy, a similar effect is expected to be observed in the
compositions of the invention, with an enhancement in the chemotherapeutic effect of
aspirin being provided by glycerin triacetate. The present compositions may therefore
provide aspirin as a chemotherapeutic agent with enhanced efficacy when compared
with the administration of aspirin alone, especially for the treatment of glioma. The
aspirin is stable and solubilised in the compositions and is therefore much more easily
administered with fewer side-effects.
Saccharin based compounds have been reported to bind to and deactivate carbonic
anhydrase IX which is found in several aggressive cancers. Saccharin based
compounds have recently been proposed as a new class of anti-cancer agent (Mahon
et al., Saccharin: a lead compound for structure-based drug design of carbonic
anhydrase IX inhibitors. Bioorg Med Chem 2015 Feb 15;23(4):849-54, incorporated
herein by reference). Proescholdt e al. ('Function of carbonic anhydrase IX in
glioblastoma multiforme', Neuro Oncol, 2012, Vol. 14, pp. 1357-1366) suggest that
inhibition of carbonic anhydrase IX is a potential metabolic target for the treatment of
glioblastoma patients.
Cardiovascular diseases or disorders that may be treated or prevented include angina
pectoris; heart failure (HF); left or right ventricular failure; pulmonary heart disease;
ischaemic heart disease (IHD); cardiomyopathy; cardiac dysrhythmia; stenosis of a
heart valve; hypertrophic cardiomyopathy (HCM); and coronary heart disease.
In some embodiments, the treatment is treatment of angina pectoris (also known as
angina), for example, angina pectoris caused by coronary heart disease; angina
pectoris caused by ischaemia; severe angina pectoris; or unresponsive or refractory
angina pectoris.
In some embodiments, the treatment is treatment of heart failure (HF), for example,
heart failure caused by ischaemia; congestive heart failure; chronic heart failure;
moderate heart failure; systolic heart failure; diastolic heart failure; or diastolic heart
failure with left ventricular injury.
In some embodiments, the treatment is treatment of left or right ventricular failure, for
example, of various aetiologies.
In some embodiments, the treatment is treatment of pulmonary heart disease, for
example, pulmonary heart disease caused by pulmonary hypertension; pulmonary
heart disease caused by chronic obstructive lung disease; or pulmonary heart disease
caused by emphysema.
In some embodiments, the treatment is treatment of ischaemic heart disease (IHD), for
example, ischaemic heart disease caused by coronary heart disease; ischaemic heart
disease caused by obstruction of the coronary artery; ischaemic heart disease caused
by spasm of the coronary artery; severe ischaemic heart disease (e.g., in a patient
awaiting coronary revascularisation); or refractory ischaemic heart disease (e.g., in a
patient with ischaemic symptoms refractory to other therapeutic measures).
In some embodiments, the treatment is treatment of cardiomyopathy, including, for
example, cardiomyopathy due to ischaemic heart disease; or cardiomyopathy due to
hypertension.
In some embodiments, the treatment is treatment of cardiac dysrhythmia (also known
as cardiac arrhythmia or irregular heartbeat), for example, cardiac dysrhythmia caused
by ischaemia.
In some embodiments, the treatment is treatment of stenosis of a heart valve, for
example, aortic stenosis, for example, inoperable aortic stenosis.
In some embodiments, the treatment is treatment of hypertrophic cardiomyopathy
(HCM), for example, symptomatic non-obstructive hypertrophic cardiomyopathy.
In some embodiments, the treatment is treatment of coronary heart disease.
In some embodiments, the treatment is of paediatric cardiovascular disease, for
example Kawasaki disease.
In some embodiments, the treatment is of congenital heart disease.
Cerebrovascular disorders or disorders that may be treated or prevented include
stroke (e.g. thrombotic or embolic stroke), cerebral ischeamia, brain ischaemia,
transient ischaemic attack (TIA) and vascular dementia.
In some embodiments, the treatment is treatment of stroke (e.g. thrombotic or embolic
stroke), transient ischaemic attack (TIA) or vascular dementia.
In some embodiments, the treatment is treatment of stroke (e.g. thrombotic or embolic
stroke), cerebral ischemia or brain ischaemia.
In some embodiments, the treatment is treatment of vascular dementia.
A cancer may be any unwanted cell proliferation (or any disease manifesting itself by
unwanted cell proliferation), neoplasm or tumor or increased risk of or predisposition to
the unwanted cell proliferation, neoplasm or tumor. The cancer may be benign or
malignant and may be primary or secondary (metastatic). A neoplasm or tumor may
be any abnormal growth or proliferation of cells and may be located in any tissue.
Examples of tissues include the adrenal gland (cortex and/or medulla), , anus,
appendix, bladder, blood, bone, bone marrow, brain, breast, caecum (and/or any part
of the large intestine), central nervous system (including or excluding the brain)
cerebellum, duodenum and/or any other part of the small intestine, epithelial cells,
gallbladder, oesophagus, glial cells, heart, kidney, lacrimal gland, larynx, liver, lung,
lymph, lymph node, lymphoblast, maxilla, mediastinum, mesentery, myometrium,
nasopharynx, omentum, oral cavity, ovary, pancreas, parotid gland, peripheral nervous
system, peritoneum, pleura, prostate, salivary gland, , skin, , soft tissues, spleen,
stomach, testis, thymus, thyroid gland, tongue, tonsil, trachea, uterus (endo- and
myometrium), cervix uteri, vulva, blood cells (leukoid and/or myeloid lines).
Cancers that may be treated or prevented include acute myeloid leukaemia; adrenal
gland cancer; biliary tract cancer; bladder cancer; bone cancer; bowel cancer; brain
cancer; breast cancer; colon cancer; colorectal cancer; endometrial cancer;
gastrointestinal cancer; genito-urinary cancer; glioma; glioblastoma; gynaecological
cancer; head cancer; Hodgkin's disease; Kaposi's sarcoma; kidney cancer; large
bowel cancer; leukaemia; liver cancer; lung cancer; lymphoma; lymphocytic leukaemia
(lymphoblastic leukaemia); malignant melanoma; mediastinum cancer; melanoma;
myeloma; myelogenous leukaemia (myeloid leukaemia); nasopharyngeal cancer; neck
cancer; nervous system cancer; non-Hodgkin's lymphoma; non-small cell lung cancer;
oesophagus cancer; osteosarcoma; ovarian cancer; pancreatic cancer; prostate
cancer; rectal cancer; renal cell carcinoma; sarcoma; skin cancer; small bowel cancer;
small cell lung cancer; soft tissue sarcoma; squamous cancer; stomach cancer;
testicular cancer; and thyroid cancer.
In some embodiments, the treatment is treatment of cancer of the gastro-intestinal
tract.
Tumors to be treated may be nervous or non-nervous system tumors. Nervous system
tumors may originate either in the central or peripheral nervous system, e.g. glioma,
gliobastoma multiforme, medulloblastoma, meningioma, neurofibroma, ependymoma,
Schwannoma, neurofibrosarcoma, astrocytoma and oligodendroglioma. Tumors may
be of the central nervous system, e.g. brain tumors, and may be primary or secondary
(metastatic). Non-nervous system cancers/tumors may originate in any other nonnervous
tissue, examples include melanoma, mesothelioma, lymphoma, myeloma,
leukemia, Non-Hodgkin's lymphoma (NHL), Hodgkin's lymphoma, chronic
myelogenous leukemia (CML), acute myeloid leukemia (AML), myelodysplasia
syndrome (MDS), cutaneous T-cell lymphoma (CTCL), chronic lymphocytic leukemia
(CLL), hepatoma, epidermoid carcinoma, prostate carcinoma, breast cancer, lung
cancer , colon cancer, ovarian cancer, pancreatic cancer, thymic carcinoma, NSCLC,
haematologic cancer and sarcoma.
In some embodiments, the treatment is of cancer of the central nervous system, e.g.
brain. The cancer to be treated may be a primary cancer or tumor. In some preferred
embodiments the cancer is a glioma, glioblastoma multiforme, high grade glioma,
diffuse intrinsic pontine glioma, medulloblastoma, meningioma, neurofibroma,
ependymoma, Schwannoma, neurofibrosarcoma, astrocytoma or oligodendroglioma.
Subjects
The subject to be treated may be any animal or human. The subject is preferably
mammalian, more preferably human. The subject may be a non-human mammal, but
is more preferably human. The subject may be male or female. The subject may be a
patient. A subject may have been diagnosed with a cardiovascular disease,
cerebrovascular disease or a cancer, or be suspected of having a cardiovascular
disease, cerebrovascular disease or a cancer. In some embodiments the subject may
be an adult, i.e. of the age 18 or over. In some embodiments a subject may be a child,
e.g. a subject under the age of 18, under the age of 16, or under the age of 12.
Routes of Administration
The liquid pharmaceutical composition comprising the salicylate compound may be
administered to a subject by any convenient route of administration, whether
systemically/peripherally or topically (i.e., at the site of desired action).
Routes of administration include, but are not limited to, oral (e.g., by ingestion); buccal;
sublingual; transdermal (including, e.g., by a patch, plaster, etc.); transmucosal
(including, e.g., by a patch, plaster, etc.); intranasal (e.g., by nasal spray, drops or from
an atomiser or dry powder delivery device); ocular (e.g., by eyedrops); pulmonary
(e.g., by inhalation or insufflation therapy using, e.g., an aerosol or vapour,
e.g., through the mouth or nose); rectal (e.g., by suppository or enema); vaginal
(e.g., by pessary); parenteral, for example, by injection, including subcutaneous,
intradermal, intramuscular, intravenous, intraarterial, intracardiac, intrathecal,
intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal, intratracheal,
subcuticular, intraarticular, subarachnoid, and intrasternal; by implant of a depot or
reservoir, for example, subcutaneously or intramuscularly.
In one preferred embodiment, the route of administration is oral (e.g., by ingestion).
In one preferred embodiment, the route of administration is parenteral (e.g. , by
injection).
In one preferred embodiment, the route of administration is rectal.
In one preferred embodiment, the route of administration is nasogastric.
In one preferred embodiment, the route of administration is pulmonary (e.g. by
inhalation or insufflation therapy using e.g. an aerosol or vapour through e.g. the
mouth or nose).
The invention provides a composition which is easily administered e.g. by the patient
themselves orally or by inhalation or insufflation. Furthermore the liquid composition
may be more easily administered to a patient in need thereof who may be unable to
accept a solid formulation.
Dosage
It will be appreciated by one of skill in the art that appropriate dosages of the
compositions comprising the salicylate compound can vary from patient to patient.
r Q† r -i h ootima! dosaoe wi!! Qenera!!v involve h balancin of h !eve! of
therapeutic benefit against any risk or deleterious side effects. The selected dosage
level will depend on a variety of factors including, but not limited to, the activity of the
particular salicylate compound, the route of administration, the time of administration,
the rate of excretion of the salicylate compound, the duration of the treatment, other
drugs, compounds, and/or materials used in combination, the severity of the condition,
and the species, sex, age, weight, condition, general health, and prior medical history
of the patient. The amount of salicylate compound and route of administration will
ultimately be at the discretion of the physician, veterinarian, or clinician, although
generally the dosage will be selected to achieve local concentrations at the site of
action which achieve the desired effect without causing substantial harmful or
deleterious side-effects.
Administration can be effected in one dose, continuously or intermittently (e.g., in
divided doses at appropriate intervals) throughout the course of treatment. Methods of
determining the most effective means and dosage of administration are well known to
those of skill in the art and will vary with the formulation used for therapy, the purpose
of the therapy, the target cell(s) being treated, and the subject being treated. Single or
multiple administrations can be carried out with the dose level and pattern being
selected by the treating physician, veterinarian, or clinician.
In general, a suitable dose of aspirin for the treatment of the disorders described
herein is in the range of about 1-30 mg per kilogram body weight of the subject per
day. Where the compound is a salt, an ester, an amide, a prodrug, or the like, the
amount administered is calculated on the basis of the parent compound and so the
actual weight to be used is increased proportionately.
For the compositions according to the invention an improved bioavailability and
absorption of the salicylate compound (e.g . aspirin) may be observed. Therefore, a
lower dose that necessary for standard compositions may be expected to achieve the
same level of therapeutic effect.
For the treatment or prevention of cardiovascular conditions, a suitable dose is typically
about 75mg or less, e.g. one of about 10 to 75mg, 10 to 65mg, 10 to 55mg, 10 to
45mg, or 10 to 20mg. Formulations may be of any kind , including for oral
administration. Administration of the dose may be once daily.
For the treatment of pain, e.g. headache, a suitable dose is typically in the range 300
to 600mg, e.g. one of about 300mg, about 325mg, about 350mg, about 400mg, about
450mg , about 500mg, about 550mg or about 600mg. Formulations may be of any kind,
including for oral administration.
For the treatment of cancer, a suitable dose is typically about 4g or less, based on the
maximum tolerated dose of aspirin for human patients being about 4g. For example,
each dose may be one of about 1g, 1.5g, 2g, 2.5g, 3g or 4g. In some preferred
embodiments, the dose may be in the range 1.5 to 3g, or 2 to 2.5g or 2 to 2.4g . In
some preferred embodiments, the dose may be one of about 2g, 2.1g, 2.2g, 2.3g, 2.4g
or 2.5g. Formulations may be of any kind , including for oral administration or
intravenous infusion. In preferred embodiments for the treatment of cancer,
formulation may be for intravenous infusion.
Packaging and Kits
One aspect of the invention pertains to a kit comprising (a) a liquid composition as
described herein, e.g., preferably provided in a suitable container and/or with suitable
packaging; and (b) instructions for use, e.g., written instructions on how to administer
the composition.
The kit may have at least one container having a predetermined quantity of the liquid
composition, e.g. predetermined dose or volume.
In one embodiment, the kit further comprises one or more (e.g., , 2, 3, 4) additional
therapeutic agents, as described herein.
The written instructions may also include a list of indications for which the active
ingredient is a suitable treatment.
In some embodiments the kit may also contain apparatus suitable to administer one or
more doses of the liquid composition, such apparatus preferably being provided in
sterile form.
In some embodiments, the kit may include packaging manufactured by the Blow-Fill-
Sea! (BFS) method.
In some embodiments a method of manufacturing a kit according to the present
invention may include the step of preventing water infiltration into the composition.
The liquid composition may be packaged into bottles for dispensing with a spoon or a
pipette. Alternatively the composition may be packaged into a syringe, vial, or in a
sachet or 'stick shot', such as a laminate 'stick shot'. The composition is suitably
packaged in individual 5 to 10 ml, preferably 5 ml servings.
The syringe may be a pre-filled syringe.
The liquid composition may also be incorporated in liquid gel capsules, e.g. in a dose
of 37.5 mg per capsule or in a dose of one of 10-50 mg per capsule, 20-40 mg per
capsule, or 30-40 mg per capsule. This preparation would be particularly applicable to
long term cardiovascular, cerebrovascular and cancer prevention uses.
Methods according to the present invention may be performed, or products may be
present, in vitro, ex vivo, or in vivo. The term "in vitro" is intended to encompass
experiments with materials, biological substances, cells and/or tissues in laboratory
conditions or in culture whereas the term "in vivo" is intended to encompass
experiments and procedures with intact multi-cellular organisms. "Ex vivo" refers to
something present or taking place outside an organism, e.g. outside the human or
animal body, which may be on tissue (e.g. whole organs) or cells taken from the
organism.
All of the features described herein may be combined with any of the above aspects of
the invention, in any combination. In addition, any upper or lower quantity or range
limit used herein may be independently combined.
The invention includes the combination of the aspects and preferred features
described except where such a combination is clearly impermissible or expressly
avoided.
The section headings used herein are for organizational purposes only and are not to
be construed as limiting the subject matter described.
Aspects and embodiments of the present invention will now be illustrated, by way of
example, with reference to the accompanying figures. Further aspects and
embodiments will be apparent to those skilled in the art. All documents mentioned in
this text are incorporated herein by reference.
Throughout this specification, including 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 or step or
group of integers or steps but not the exclusion of any other integer or step or group of
integers or steps.
It must be noted that, as used in the specification and the appended claims, the
singular forms "a," "an," and "the" include plural referents unless the context clearly
dictates otherwise. Ranges may be expressed herein as from "about" one particular
value, and/or to "about" another particular value. When such a range is expressed,
another embodiment includes from the one particular value and/or to the other
particular value. Similarly, when values are expressed as approximations, by the use
of the antecedent "about," it will be understood that the particular value forms another
embodiment.
Brief Description of the Figures
Embodiments and experiments illustrating the principles of the invention will now be
discussed with reference to the accompanying figures in which:
Figure 1 is a plot of the normalized % of salicylic acid present in a test composition
against the number of days elapsed since preparation of the composition, for a
composition of 90 wt% glycerol+20EO and 10 wt% aspirin. The test was conducted at
25°C.
Figure 2 is a plot of the normalized % of salicylic acid present in a test composition
against the number of days elapsed since preparation of the composition, for a
composition of 89 wt% giyceroi -20EO, 10 wt% aspirin and 1 w % saccharin. The test
was conducted at 25°C.
Figure 3 is a plot of the normalized % of salicylic acid present in a test composition
against the number of days elapsed since preparation of the composition, for a
composition of 96.5 wt% glycerin triacetate, 2.5 wt% aspirin and 1 wt% saccharin. The
test was conducted at 25°C.
Figures 4A to 4E. Charts showing percentage cell death with varying concentration
of drug (liquid aspirin (ASP); triacetin alone (ASP vehicle); temozoiomide alone (TMZ))
in ex vivo patient glioblastoma cell lines (A) SEBTA 023, (B) SEBTA 003, (C) UP 029,
(D) KNS42, (E) SNF188. Studies were conducted at 96 hours post-drug treatment and
show the average (+/- StDev) of three experiments conducted in triplicate. Due to
solubility issues, aspirin alone was not included in this study due to an inability to treat
at comparable concentrations.
Figure 5. Representative microscopy images of treated ex vivo biopsy-derived cell
lines. Images show non-treated (NT), liquid aspirin ("aspirin") and triacetin only
treatment. Contrast differences are the result of cell media pH changes following drug
treatment and not due to cell confluence variation.
Figures 6A to 6C. (A) Chart showing percentage cell death in ex vivo biopsyderived
UP029 cells with varying concentration of drug (saccharin-only (Sac), liquid
aspirin (Lqd Aspirin), triacetin-only, temozolomide-only (TMZ)). (B) and (C) Charts
showing synergy analysis of ex vivo biopsy-derived UP029 cells following treatment
with liquid aspirin, saccharin-only and triacetin-only. Synergy (of the liquid aspirin
compound) was noted (in order to kill 30%, 50% and 70%) compared to each agent
alone with the exception of triacetin-alone (at 90% reduced cell viability) at 96 hours
post-treatment, where the highest dosages of triacetin alone was reducing overall cell
viability.
Figures 7(A) to 7(F). Charts showing percentage cell death (MTS viability analysis) of
control and biopsy-derived cell lines 96 hours post-treatment with varying
concentration of drug (saccharin-only (Sac), liquid aspirin-only (Lqd Aspirin), triacetinonly,
or temozolomide-only (TMZ)). (A) Non-neoplastic astrocytes (CC2565) included
as a control, (B) SEBTA 003 (C) SEBTA 023 (D) KNS42 (E) SNF188 (F) SEBTA 025.
Each plot is representative of 3 studies conducted in triplicate.
In this specification the following test method was used:
Aspirin Stability
Aspirin undergoes hydrolysis to salicylic acid and acetic acid. The aspirin and salicylic
acid concentrations in the sample composition were determined for a minimum period
of at least 200 days, preferably up to a maximum of 300 days. The composition was
stored in a sealed glass vial in an oven at 25°C, and the concentration of aspirin and
salicylic acid measured weekly. The glass vial was opened, a sample removed for
testing every week and the glass vial resealed after purging with nitrogen. High
performance liquid chromatography with UV detection was used. The conditions were
as follows:
• Mobile phase: 40% of 1% acetic acid in water, 60% methanol.
• Column: Agilent Zorbax Eclipse XBD-C18. 4.6 mm x 50 mm with 5 micron particle
size.
• Column heater: 25°C.
• Sample concentration: 0.02 g made to 10 ml with mobile phase.
• Injection volume: 40 microlitre.
• Flow rate: 1 ml minute.
· Detection: UV at 280 nm.
The stability of the aspirin in the composition is defined as the aspirin degradation rate
which was calculated as (i) the average % aspirin degradation (based on the original
aspirin concentration) per day, and (ii) the average % aspirin degradation (based on
the original aspirin concentration) per gram of composition per day.
The invention is illustrated by the following non-limiting examples.
Examples
Example 1
Food grade glycerin triacetate (ex Sigma Aldrich) was mixed with ethanoi 40% w/v and
passed through a fixed bed of activated earth. The solvent was then removed using
vacuum distillation followed by steam distillation to levels below 1 ppm of ethanoi in the
glycerin triacetate.
Example 2
A composition was prepared by mixing 2.5 wt% aspirin (ex Sigma Aldrich), 96.5 wt%
glycerin triacetate (produced in Example 1), and 1 wt% saccharin (ex Sigma Aldrich).
The components were mixed in the appropriate ratios and sonicated to achieve
complete solution. Microscopy showed that the solution was free of any particulate
material. The aspirin stability in the composition was measured weekly as described
above. The aspirin degradation after 277 days was 6.9% of the original amount
present, which is equivalent to a degradation rate of 0.025%/day.
Example 3
The procedure of Example 2 was repeated except that the composition additionally
contained 0.15 wt% of spearmint oil (ex Quinessence) (and correspondingly 0.15 wt%
less of glycerin triacetate, i.e. 96.35 wt%). The aspirin degradation after 246 days was
5.7% of the original amount present, which is equivalent to a degradation rate of
0.023%/day.
Example 4
A composition was prepared by mixing 2.5 wt% aspirin (ex Sigma Aldrich), 96.5 wt%
glycerin triacetate (produced in Example 1), and 1 wt% saccharin (ex Sigma Aldrich).
The components were mixed in the appropriate ratios and sonicated to achieve
complete solution. Microscopy showed that the solution was free of any particulate
material. The aspirin stability in the composition was measured weekly as described
above.
The "normalised % salicylic acid" was calculated by finding the % salicylic acid in the
composition, as a percentage of the total % salicylic acid and aspirin in the
composition, according to the formula
Normalised % salicylic acid = (%salicylic acid)/(%aspirin + %salicylic acid)
The results are shown in Table 1 below, and in Figure 3 .
Table 1
The observed 5.35% degradation after 186 days is equivalent to a degradation rate of
0.0288%/day.
Extrapolating from these results, the predicted shelf life limit (the point in time after
initial preparation at which the normalised % salicylic acid reaches 10%) is 360 days.
Example 5
Compositions of aspirin with glycerin triacetate and saccharin were prepared according
to the method of Example 2 . Five compositions were prepared, with aspirin
concentrations of 2.5 wt%, 3.0 w†%, 3.5 wt%, 4.0 wt% and 4.5 wt% respectively. Each
composition also contained 1 wt% saccharin and the balance glycerin triacetate.
The compositions were tested for solubility at different temperatures. Compositions
were observed to determine whether the aspirin had fully dissolved to provide a clear
solution and whether the solution remained clear.
The results are shown in Table 2 below:
Table 2
The results demonstrate stable solubilisation of aspirin in the composition of the
present invention at 4 wt% concentration at 25°C. At a concentration of 2.5 wt% the
aspirin was stably solubilised at temperatures down to 4°C.
The compositions of the invention are therefore particularly suitable for use in the
treatment of cardiovascular and cerebrovascular disorders and cancer, where low
doses of aspirin over extended periods are required.
For example, at 2.5 wt% aspirin, a 2 mL dose in a gel capsule would provide around 50
mg aspirin. A 1 mL dose would provide around 25 mg aspirin.
Comparative Example 1
A composition was prepared by mixing 10 wt% aspirin (ex Sigma Aldrich) and 90 wt%
glycerol+20EO (glycerol ethoxylated with 20 mol equivalents of ethylene oxide). The
components were mixed in the appropriate ratios and sonicated to achieve complete
solution. Microscopy showed that the solution was free of any particulate material.
The aspirin stability in the composition was measured weekly as described above.
The results are shown in Table 3 below, and in Figure 1.
Table 3
The average degradation rate is 0.102%/day. Extrapolation from these results gives a
predicted shelf life of 124 days.
Comparative Example 2
A composition was prepared by mixing 0 wt% aspirin (ex Sigma Aldrich), 1%
saccharin (ex Sigma Aldrich) and 89 wt% glycerol+20EO. The components were
mixed in the appropriate ratios and sonicated to achieve complete solution.
Microscopy showed that the solution was free of any particulate material. The aspirin
stability in the composition was measured weekly as described above.
The results are shown in Table 4 below, and in Figure 2.
Table 4
Day Normalised %salicylic acid
0 0.8
8 1.4
14 1.1
2 1 1.6
28 2.0
35 2.5
The average degradation rate is 0.071 4%/day. Extrapolation from these results gives
a predicted shelf life of 205 days.
The results of the above Examples and Comparative Examples are summarised in
Table 5 below:
Table 5
It can be seen that the degradation rate of aspirin and thereby the shelf-life of the liquid
composition is improved dramatically for compositions according to the invention.
Shelf life of a year or longer is possible. A comparison of Comparative Examples 1
and 2 shows that the presence of saccharin provides a significant increase in stability.
Furthermore, a comparison of Comparative Example 2 with Examples 2-4 reveals a
dramatic improvement in stability when the composition includes glycerin triacetate.
The above examples illustrate the improved properties of a composition according to
the present invention.
Example 6 - An in vitro study to investigate the anti-tumour effects of liquid
aspirin in adult glioblastoma, paediatric high grade glioma and medulloblastoma
Introduction
Soluble aspirins currently on the market are in fact dispersible and therefore still
contain grains that sit on the gastric mucosa causing gastric side effects. The
categorization as "soluble" is therefore not accurate. Alternative aspirin products are
powders that quickly disperse in water. No truly shelf stable liquid formulation of
acetylsalicylic acid (ASA) has been successfully produced, until now. The unique liquid
ASA described herein (referred to in this Example as 'liquid aspirin') is expected to
show a significant reduction in gastrointestinal side effects.
As described herein, liquid aspirin contains ASA and two excipients: glycerin triacetate
(triacetin) and saccharin (Sac). All three ingredients are pharmaceutically approved
and have been shown to have compelling anti-tumour properties.
Triacetin has been shown to significantly augment drug delivery across the blood brain
barrier (BBB), suggesting that this combination could be highly effective against
glioblastoma (GBM) [Van Tel!ingen et al., Overcoming the blood-brain tumor barrier for
effective glioblastoma treatment. Drug Resist. Updat. 19, 1-12 (2015)].
Saccharin based compounds have been proposed as a new class of anti-cancer agent
(Mahon et al., Saccharin: a lead compound for structure-based drug design of carbonic
anhydrase IX inhibitors. Bioorg Med Chem 2015 Feb 15;23(4):849-54, incorporated
herein by reference). Proescholdt et al. ('Function of carbonic anhydrase IX in
glioblastoma multiforme', Neuro Oncol, 2012, Vol. 14, pp. 1357-1366) suggest that
inhibition of carbonic anhydrase IX is a potential metabolic target for the treatment of
glioblastoma patients.
Of the three components, aspirin has demonstrated the most potent anti-tumour effect,
particularly against GBM. An initial in vivo study highiighied that the administration of
aspirin into an established Fischer 344 rat glioma model (Aas.A.T., Brun.A.,
Blennow.C, Stromblad.S., & Salford.L.G. The RG2 rat glioma model. J. Neurooncol.
23, 175-183 (1995)) significantly inhibited the growth of differentiated malignant
glioblastoma RG2 cells both when administered the day before tumour cell inoculation
as well as in established rat glioblastoma tumours (Aas.A.T., Tonnessen.T.I., Brun.A.,
& Salford.L.G. Growth inhibition of rat glioma cells in vitro and in vivo by aspirin. J.
Neurooncol. 24, 7 1- 80 ( 1995)).
Prostaglandin E2 (PGE2) has been shown to have an important role in both
immunosuppression and tumour growth. As a PGE2 inhibitor, aspirin has been shown
to reduce in vitro tumour cell proliferation. Aspirin dosage studies were conducted,
evaluating the effect of both high and low dose aspirin exposure on PGE2 synthesis in
the in vitro C6 glioma model. These studies revealed that aspirin directly inhibited
PGE2 synthesis in C6 cells and that critically, low-dose aspirin is as effective as highdose
aspirin in mediating this response (Hwang,S.L. et al. Effect of aspirin and
indomethacin on prostaglandin E2 synthesis in C6 glioma cells. Kaohsiung. J. Med.
Sci. 20, 1-5 (2004)).
Human A172 glioblastoma cells treated with aspirin induced significant apoptosis
(programmed cell death) [Kim.S.R. et al. Aspirin induces apoptosis through the
blockade of IL-6-STAT3 signaling pathway in human glioblastoma A 72 cells.
Biochem. Biophys. Res. Commun. 387, 342-347 (2009)]. The underlying mechanism
for this response was a reduction in the level of phosphorylated signal transducer and
activator of transcription 3 (STAT3), specifically pTyr-STAT3. STAT3 is a transcription
factor that is required for survival of A172 cells. This conclusion was supported by
measuring cyclin D1, XIAP, and Bcl-2 transcription that was notably attenuated after
aspirin treatment (Kim et al., supra). Implicating STAT3 further, the expression and
secretion of interleukin-6 (IL-6) (that induces STAT3 phosphorylation), was notably
inhibited by aspirin treatment (Kim et al., supra).
Drawing from these findings, it is known that hypoxia can activate STAT3 and
subsequently induce angiogenesis (the development of blood vessels) [Greten.F.R. &
Karin.M. Peering into the aftermath: JAKi rips STAT3 in cancer. Nat. Med. 16, 1085-
087 (2010)]. In most solid malignancies, persistent STAT3 signalling is triggered by
an autocrine-paracrine production of IL-6 that is noticeably higher in a hypoxic
environment (Song.Y.Y. et al. STAT3, p-STAT3 and HIF-1alpha are associated with
vasculogenic mimicry and impact on survival in gastric adenocarcinoma. Oncol. Lett. 8,
431-437 (2014)). Hypoxia is a hallmark of GBM, with tumours showing
pseudopalisades of neoplastic cells surrounding areas of frank necrosis as well as
signs of vascular proliferation. These areas of peri-necrotic hyper-cellularity have been
well characterized and are not the result of increased proliferation. As one would
predict, these regions have high levels of hypoxia-induced factor 1 alpha (HIF1a)
expression, resulting in pro-angiogenic vascular endothelial growth factor (VEGF)
secretion as well as elevated IL-6. This in turn drives vascular proliferation. However,
the vessels that are generated in response to VEGF within this environment are
severely malformed (Jain.R.K. Normalizing tumor microenvironment to treat cancer:
bench to bedside to biomarkers. J. Clin. Oncol. 31, 2205-2218 (2013)). The result is
deregulated vessel structure with gaps between endothelial cells and an absence of
pericytes. Due to this malformation and inherent leakiness, the interstitial pressure is
increased resulting in vascular stasis with corresponding exacerbation of hypoxia and
increased microvascular thrombosis (Jain et al., supra). Strikingly, it has been shown
that aspirin selectively suppresses inflammation, and specifically IL-6-induced T-helper
cell 7. This mediates the down regulation of acetyl-STAT3 expression as well as
blocking IL-17A-induced inflammation and IL-6 production. This reduction of IL-6
production will then result in a concomitant reduction in active (phosphorylated)
STAT3.
More recently, it has been shown that aspirin represses the transcriptional activity of
the b-catenin/TCF protein complex. As a consequence of this, aspirin directly inhibits
GBM proliferation and invasion as well as inducing apoptosis (Jin.T., George, F., I , &
Sun, . nt and beyond Wnt: multiple mechanisms control the transcriptional property
of beta-catenin. Cell Signal. 20, 1697-1704 (2008)). The results presented within this
study suggest that aspirin exerts its anti-neoplastic action by suppressing the b-
catenin/TCF signalling pathway in GBM. This is particularly striking as recent data has
highlighted that FoxM1 promotes the development and progression of GBM by
regulating key factors involved in cell division, epithelial to mesenchymal transition
(EMT), invasion, angiogenesis and upregulation of the Wnt/p-catenin signalling
network (Wang.Z., Zhang, S., Siu.T.L, & Huang,S. Glioblastoma multiforme formation
and EMT: roie of FoxM transcription factor. Curr. Pharm. Des 21, 1268-1271 (2015)).
A deregulated Wni/p-catenin network has been reported in GBM and it has been
suggested that this could also constitute a therapeutic target (Zhang, K. et al. ICAT
inhibits glioblastoma cell proliferation by suppressing Wnt/beta-catenin activity. Cancer
Lett. 357, 404-41 1 (2015)).
Experimental Method and Results
Many of the established glioma cell lines, such as C6, A172, U87, U373 and U251
have been grown/passaged in research laboratories around the world for many years
and, as a consequence, display a high degree of cellular heterogeneity rendering them
increasingly dissimilar to their original primary/early passage cultures and, indeed,
from the biopsy from which they were derived.
To address this issue, we used a panel of patient-derived (adult and paediatric) ex vivo
GBM low passage cell cultures, which have been characterized at the molecular level,
including DNA fingerprinting (Prof. G. Pilkington, Brain Tumour Research Centre,
University of Portsmouth, UK). These cells are truly representative of the patient GBM
and, as a result, any anti-tumour effect observed will have clinical relevance. This
extensive cell culture bio-bank allows screening of novel anti-GBM therapeutics prior to
in vivo studies and clinical trials.
It is also critical to compare these novel therapeutics against the currently prescribed
frontline chemotherapeutics. Consequently, our studies also include temozolomide,
which is a standard of care treatment for GBM.
Non-neoplastic astrocytes provide a control element within these studies. These are
non-cancerous and any proposed therapeutic will preferably have selective anti-cancer
action (i.e. not kill normal, non-tumor, cells).
Viability studies were conducted to compare the effect of liquid aspirin (ASP), triacetin
(ASP vehicle), and temozolomide (TMZ) in five adult GBM ex vivo cell lines from our
panel. Results are shown in Figures 4A to 4E. Liquid aspirin showed notably better
induction of cell death in all GBM cells tested.
Synergy studies were also conducted to directly address the specific anti-tumour
efficacy of each of aspirin, triacetin and saccharin (Sac) as individual components and
the triple-formulation of aspirin, triacetin and saccharin (liquid aspirin). We have
significant experience conducting this type of analysis and importantly can differentiate
between additive effects versus a synergistic response (Hallden.G. et al. Novel
immunocompetent murine tumor models for the assessment of replication-competent
oncolytic adenovirus efficacy. Mol. Ther. 8, 412-424 (2003); Cheong,S.C. et al. E1Aexpressing
adenoviral E3B mutants act synergistically with chemotherapeutics in
immunocompetent tumor models. Cancer Gene Ther. 15, 40-50 (2008)). Figures 6B
and 6C show synergy of liquid aspirin in order to kill 30%, 50% and 70% of UP 029
cells compared to use of each agent alone with the exception of triacetin-alone at 90%
reduced cell viability, where the highest dosages of triacetin alone reduced overall cell
viability.
We also confirmed that liquid aspirin demonstrates anti-cancer specificity, i.e. is not
toxic to non-neoplastic astrocytes. Results are shown in Figure 7A to 7F which show
liquid aspirin to have markedly better cell killing ability than triacetin or temozolomide;
liquid aspirin typically having at least one order of magnitude greater potency in
inducing cell death than triacetin or temozolomide. Liquid aspirin had significantly less
toxicity when added to non-neoplastic cells (Figure 7A). Indeed, the associated toxicity
noted within the astrocyte cell line could be associated with triacetin-alone (which
demonstrated a similar MTS-viability profile).
References
The following are incorporated herein by reference:
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CLAIMS
1. A liquid composition comprising a salicylate compound, a glycerol derivative,
and a saccharin compound.
2 . The composition according to claim 1 further comprising a flavouring agent.
3 . The composition according to claim 1 or 2 wherein the salicylate compound is
selected from the group consisting of: aspirin, triflusal, diflunisal, salsalate and salicylic
acid.
4 . The composition according to any one of claims 1 to 3 wherein the salicylate
compound is aspirin.
5. The composition according to any one of claims 1 to 4 wherein the glycerol
derivative is glycerin triacetate.
6. The composition according to any one of claims 1 to 5 wherein the saccharin
compound is saccharin.
7. The liquid composition according to any one of claims 1 to 6 wherein the
composition comprises or consists of a salicylate compound, glycerin triacetate, and
saccharin.
8. The composition according to any one of claims 1 to 7 wherein the composition
comprises or consists of aspirin, glycerin triacetate, and saccharin.
9. The composition according to any one of claims 1 to 8 wherein the composition
comprises or consists of aspirin, glycerin triacetate, saccharin and a flavouring agent.
10. The composition according to any one of the preceding claims wherein the
flavouring agent comprises or consists of mint oil.
11. The composition according to any one of the preceding claims, wherein the
concentration of the salicylate compound is 0.5 to 3% and/or the concentration of the
glycerol derivative, preferably glycerin triacetate, is 94 to 99%.
2. The composition according to any one of the preceding claims, wherein the
concentration of saccharin compound, preferably saccharin, is 0.1 to 3%.
13. The composition according to any one of the preceding claims wherein the
glycerol derivative, preferably glycerin triacetate, is obtainable by passing through
activated earth.
14. The composition according to any one of the preceding claims having a
salicylate compound degradation rate at 25°C of less than 0.04%/day and/or less than
0.006 mg/g/day.
15. The composition according to any one of the preceding claims having a
salicylate compound degradation rate at 25°C of less than 0.02%/day and/or less than
0.004 mg/g/day
16. The composition according to any one of the preceding claims which is free of
particulates and/or in which the salicylate compound is completely soluble.
17. The composition according to any one of the preceding claims which is suitable
for o al use.
18. The composition according to any one of the preceding claims which is
formulated for intravenous or intra-arterial administration.
9. The composition according to any one of the preceding claims which is
formulated for inhalation or insufflation administration.
20. A method of preparing a liquid composition comprising admixing a salicylate
compound, glycerol derivative, and a saccharin compound.
2 1. The method according to claim 20 further comprising admixing a flavouring
agent.
22. The method according to claim 20 or 2 1 wherein the salicylate compound is
selected from the group consisting of: aspirin, triflusal, diflunisal, salsalate and salicylic
acid.
23. The method according to any one of claims 20 to 22 wherein the salicylate
compound is aspirin.
24. The method according to any one of claims 20 to 23 wherein the glycerol
derivative is glycerin triacetate.
25. The method according to any one of claims 20 to 24 wherein the saccharin
compound is saccharin.
26. The method according to any one of claims 20 to 25 wherein the method
comprises admixing aspirin, glycerin triacetate and saccharin.
27. The method according to any one of claims 20 to 36 wherein the flavouring
agent is mint oil.
28. The method according to any one of claims 20 to 27, wherein the concentration
of the salicylate compound is 0.5 to 3% and/or the concentration of the glycerol
derivative, preferably glycerin triacetate, is 94 to 99%.
29. The method according to any one of claims 20 to 28, wherein the concentration
of saccharin compound, preferably saccharin, is 0.1 to 3%.
30. A packaged article comprising the composition as defined in any one of claims
1 to 19 which is sealed therein.
3 . The packaged article according to claim 30 selected from the group consisting
of a bottle, pipette, syringe, vial, sachet, stick shot and liquid gel capsule.
32. A method of taking aspirin which comprises orally administering a liquid
composition according to any one of claims 1 to 19.
33. A method of taking aspirin which comprises oral, rectal, nasogastric, parenteral
preferably intravenous or intrarterial, inhalation or insufflation administration of the
liquid composition according to any one of claims 1 to 19.
34. A liquid composition according to any one of claims 1 to 19 for use in a method
of medical treatment.
35. The liquid composition for use in a method of treatment according to claim 34,
wherein the method of medical treatment involves oral, rectal, nasogastric, parenteral
preferably intravenous or intrarterial, inhalation or insufflation administration of the
liquid composition.
36. The liquid composition according to any one of claims 1 to 19, for use in a
method of treatment of a cardiovascular disease, cerebrovascular disease or cancer.
37. The liquid composition for use in a method of treatment according to claim 36,
wherein the cardiovascular disease is selected from angina pectoris; heart failure (HF);
left or right ventricular failure; pulmonary heart disease; ischaemic heart disease (IHD);
cardiomyopathy; cardiac dysrhythmia; stenosis of a heart valve; hypertrophic
cardiomyopathy (HC-M); coronary heart disease; paediatric cardiovascular disease
(e.g. Kawasaki disease); and congenital heart disease.
38. The liquid composition for use in a method of treatment according to claim 36,
wherein the cerebrovascular disease is selected from stroke, cerebral ischaemia, brain
ischaemia, transient ischaemic attack (TIA) and vascular dementia.
39. The liquid composition for use in a method of treatment according to claim 36,
wherein the cancer is a cancer occurring in the central nervous system, a brain cancer,
a gastrointestinal cancer or glioma.
40. The liquid composition according to any one of claims 1 to 19, for use in a
method of treatment of pain, fever or inflammation.