USE OF 3-CARBOXY-N-ETHYL-N,N-DIMETHYLPROPAN-1-AMINIUM OR A PHARMACEUTICALLY ACCEPTABLE SALT THEREOF IN THE PREVENTION AND TREATMENT OF DIABETES

UTILISATION DE 3-CARBOXY-N-ETHYL-N,N-DIMETHYLPROPAN-1-AMINIUM OU D'UN SEL PHARMACEUTIQUEMENT ACCEPTABLE DE CELUI-CI DANS LA PREVENTION ET LE TRAITEMENT DU DIABETE

English Abstract

Use of 3-carboxy-N-ethyl-N,N-dimethylpropan-1-aminium and its pharmaceutically acceptable salts to decrease blood plasma levels of insulin and glucose.

French Abstract

L'invention concerne l'utilisation de 3-carboxy-N-éthyl-N,N-diméthylpropan-1-aminium et de ses sels pharmaceutiquement acceptables pour diminuer les taux d'insuline et de glucose dans le plasma sanguin.

Claims

9
Claims
1. 3-Carboxy-N-ethyl-N,N-dimethylpropan-1-aminium or pharmaceutically
acceptable salts thereof for use in the prevention and treatment of diabetes.
2. 3-Carboxy-N-ethyl-N,N-dimethylpropan-1-aminium or pharmaceutically
acceptable salts for use according to claim 1, wherein the pharmaceutically
acceptable salt is 3-carboxy-N-ethyl-N,N-dimethylpropan-1-aminium hydrogen
fumarate.
3. 3-Carboxy-N-ethyl-N,N-dimethylpropan-1-aminium or pharmaceutically
acceptable salts for use according to claim 1, wherein the pharmaceutically
acceptable salt is 3-carboxy-N-ethyl-N,N-dimethylpropan-1-aminium
dihydrogen phosphate.

Description

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1
Description
Use of 3-carboxy-Abethyl-N,N-dimethylpropan-1-aminium or a pharmaceutically
acceptable
salt thereof in the prevention and treatment of diabetes
Technical Field
The present invention relates to 3-carboxy-N-ethyl-/V,N-dimethylpropan-1-
aminium or
a pharmaceutically acceptable salt thereof for use in the prevention and
treatment of
diabetes. Examples of pharmaceutically acceptable salts of 3-carboxy-N-ethyl-
/V,/V-
dimethylpropan-1-aminium are: 3-carboxy-N-ethyl-/V,N-dimethylpropan-1-aminium
hydrogen fumarate and 3-carboxy-N-ethyl-/V,/V-dimethylpropan-1-aminium
dihydrogen
phosphate.
Background Art
3-Carboxy-/V,/V,N-trimethylpropan-1-aminium (GBB) is known mostly as a bio-
precursor of carnitine, a key molecule in the regulation of myocardial energy
metabolism. It was primarily characterised as a toxic substance, which
accelerates
respiration, causes salivation and lacrimation, pupil dilation,
vasoconstriction and
cardiac arrest in diastole LINNEWEH W. Gamma- Butyrobetain, Crotonbetain und
Carnitin im tierischen Stoffwechsel. Hoppe-Seylers Zellschnft fur
physiologische
Chemie. 1929, vol.181, p.42-53. In later publications it has been shown 3-
carboxy-
N,N,N-trimethylpropan-1-aminium has extremely low toxicity (LD50 7000 mg/kg,
s.c.)

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ROTZSCH. W. Uber die Toxizitat des Carnitins und einiger verwandter Stoffe.
Acta
biol. med. germ. 1959, vol.3, p.28-36.
Methods for preparation of a new compound with cardioprotective activity - 3-
carboxy-
N-ethyl-/V,N-dimethylpropan-1-aminium has been disclosed in WO 2011/048201 A
(GRINDEKS, JSC) 28/04/2011 . Salts of 3-carboxy-N-ethyl-/V,N-dimethylpropan-1-
aminium as well as their use in the treatment of cardiovascular diseases has
been
disclosed in patent publication WO 2012/146736 A (GRINDEKS, JSC) 27/12/2012.
Diabetes mellitus is one of the leading causes of morbidity and mortality
globally, and
despite medical breakthroughs and treatment innovations, the number of adults
with
this condition has been increasing at a faster pace than previously expected
DANAEI
G., et al. National, regional, and global trends in fasting plasma glucose and
diabetes
prevalence since 1980: systematic analysis of health examination surveys and
epidemiological studies with 370 country-years and 2.7 million participants.
Lancet.
2011, vol.378, p.31-40. Type 2 diabetes mellitus (T2DM) comprises about 90% of
all
diabetes cases throughout the world. T2DM is a metabolic disorder
characterized by
elevated blood glucose levels in the context of insulin resistance and
relative insulin
deficiency. Insulin resistance contributes to hyperinsulinaemia which is often
seen in
early stages of T2DM. Furthermore, hyperinsulinaemia is predictive of
development of
T2DM, as it is known to precede T2DM by decades. T2DM causes dysfunctions in
multiple organs and tissues leading to disability and premature death of T2DM
patients. Diabetes-related expenses to healthcare systems and patients are
projected
to increase twice during next 25 years HUANG E.S., et al. Projecting the
future

3
diabetes population size and related costs for the U.S.. Diabetes Care. 2009,
vol.32,
p.2225-2229. , therefore anti-diabetes drugs are of high necessity and
economical
value.
Although several classes of anti-diabetic drugs are currently available, there
is a
significant need for novel therapies with added medical value and improved
side-effect
profile. Achieving and maintaining long-term glycaemic control by existing
drugs in
T2DM patients is often challenging, also due to insufficient insulinaemic
control
MANNINO G.C., SESTI G. Individualized Therapy for Type 2 Diabetes: Clinical
Implications of Pharmacogenetic Data. Mol Diagn Ther. 2012, vol.16, p.285-302.
,
SAFAVI M., et al. The importance of synthetic drugs for type 2 diabetes drug
discovery.
Expert Opin Drug Discov. Ahead of Print (doi:10.1517/17460441.2013.837883).
19.09.2013, p.1-25. Normalizing insulin levels is of particular preventive and
therapeutic
importance since uncontrolled insulin resistance and subsequent
hyperinsulinaemia are
major contributors in development of T2DM. Therefore, new pharmacotherapy
targeting
hyperglycaemia, insulin resistance and hyperinsulinaemia could be of
particular interest
for effective prevention and treatment of T2DM.
Summary of invention
The present invention is directed to prevention and treatment of diabetes by
decreasing
blood plasma levels of insulin and glucose.
The present invention relates to 3-carboxy-N-ethyl-N,N-dimethylpropan-1-
aminium or a
pharmaceutically acceptable salt thereof for use in the prevention and
treatment of
diabetes.
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During our research to find new active substances to treat hyperglycaemia an
unexpected glucose- and insulin- reducing effect of 3-carboxy-N-ethyl-/V,N-
dimethylpropan-1-aminium salts has been discovered.
The reduction of glucose and insulin concentration in blood is achieved by
treatment
with 3-carboxy-N-ethyl-N,N-dimethylpropan-1-aminium or its pharmaceutically
acceptable salts, for example but not limited to: 3-carboxy-N-ethyl-N,N-
dimethylpropan-1-aminium 2-(acetyloxy)benzoate, 3-carboxy-N-ethyl-/V,N-
dimethylpropan-1-aminium hydrogen fumarate, 3-carboxy-N-ethyl-/V,N-
dimethylpropan-1-aminium dihydrogen phosphate, 3-carboxy-N-ethyl-/V,N-
dimethylpropan-1-aminium 2,6-dioxo-1,2,3,6-tetrahydropyrimidine-4-carboxylate,
3-
carboxy-N-ethyl-N,N-dimethylpropan-1-aminium 3-carboxypropanoate, preferably 3-
carboxy-N-ethyl-/V,N-dimethylpropan-1-aminium hydrogen fumarate or 3-carboxy-N-
ethyl-N,N-dimethylpropan-1-aminium dihydrogen phosphate.
3-Carboxy-N-ethyl-/V,N-dimethylpropan-1-aminium can be used in pharmaceutical
preparations containing the compound, or a pharmaceutically acceptable salt
thereof,
in combination with a pharmaceutically acceptable carrier or diluent. Suitable
pharmaceutically acceptable carriers include inert solid fillers or diluents
and sterile
aqueous or organic solutions. Thus, for oral administration the compounds can
be
combined with a suitable solid or liquid carrier or diluent to form capsules,
tablets,
powders, syrups, solutions, suspensions and the like. The pharmaceutical
compositions may, if desired, contain additional components such as
flavorants,
sweeteners, excipients and the like. For parenteral administration the
compounds can

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be combined with sterile aqueous or organic media to form injectable solutions
or
suspensions. The injectable solutions can then be administered intravenously,
intraperitoneally, subcutaneously, or intramuscularly. 3-Carboxy-N-ethyl-N,N-
dimethylpropan-1-aminium or a pharmaceutically acceptable salt therefore can
also be
prepared for transdermal application, for example, in a form of adhesive
plaster.
A therapeutically effective amount of 3-carboxy-N-ethyl-/V,/V-dimethylpropan-1-

aminium or its pharmaceutically acceptable salt is about 0.01 to 500
mg/kg/day,
preferably 0.1 to 100 mg/kg/day.
Example
Female ApoE-/- mice weighing 18 - 20 g were maintained on a 12 h dark/12 h
light
cycle in air-conditioned rooms (22.5 0.5 C, 50 5% humidity) with unlimited
access to
food and water.
Mice were adapted to local conditions for one week before the beginning of the
study.
At the age of 8 weeks, mice were randomly assigned to five equally sized
groups (n =
10). Experimental animals of all groups were fed with WESTERN RD (P) diet (Cat
82316) from Special Diets Services (Great Britain) for 4 months. During these
4
months, mice from different experimental groups received following treatment:
Control group - drinking water;
GBB group - GBB 10mg/kg;
3-carboxy-N-ethyl-/V,N-dimethylpropan-1-aminium hydrogen fumarate group - 3-
carboxy-N-ethyl-N,N-dimethylpropan-1-aminium hydrogen fumarate 17.5 mg/kg;

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3-carboxy-N-ethyl-/V,/V-dimethylpropan-1-aminium dihydrogen phosphate group -
3-
carboxy-N-ethyl-/V,N-dimethylpropan-1-aminium dihydrogen phosphate 16.8 mg/kg.
GBB, 3-carboxy-N-ethyl-/V,N-dimethylpropan-1-aminium hydrogen fumarate or 3-
carboxy-N-ethyl-/V,N-dimethylpropan-1-aminium dihydrogen phosphate were added
to
the drinking water. The doses of GBB, 3-carboxy-N-ethyl-/V,N-dimethylpropan-1-
aminium hydrogen fumarate and 3-carboxy-N-ethyl-/V,/V-dimethylpropan-1-aminium
dihydrogen phosphate were equimolar to the 10 mg/kg dose (62.5 inium d4-
(ethyl(dimethyl)ammonio)butanoate. The dosing of the test compounds was
confirmed
by measuring the consumption of drinking water every 2 days and adjusting the
concentration of supplemented substances.
At the age of 8 and 16 weeks blood was collected into EDTA/diamide- and
protease
inhibitor (PMSF, pepstatin, leupeptin, aprotinin)-containing tubes. Blood
glucose
concentration was measured using a MediSense Optium Xceed blood glucose meter
and strips. Plasma was obtained by centrifugation at 3000 x g and stored
frozen (-80
C) until analysis. Plasma insulin concentrations were determined with a RIA
kit
(Biotrend, Germany).
Treatment-effect on plasma glucose level is presented in Table 1. Both 3-
carboxy-N-
ethyl-/V,N-dimethylpropan-1-aminium hydrogen fumarate and 3-carboxy-N-ethyl-
/V,N-
dimethylpropan-1-aminium dihydrogen phosphate reduced plasma glucose levels in
a
statistically significant manner, while GBB lacked an effect.

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Table 1
Effects of test compounds at a dose of 10mg/kg on plasma glucose concentration
in
fed and fasted ApoE-/- mice
Week 8 Week 16
Fed Fasted Fed Fasted
Control group 8,0 0,42 5,1 0,28 10,0 0,55
5,4 0,33
3-carboxy-N-ethyl-N,N-
dimethylpropan-1-aminium
dihydrogen phosphate group 8,2 0,61 3,7 0,18* 8,2 0,39*
4,3 0,16*
3-carboxy-N-ethyl-N,N-
dimethylpropan-1-aminium
hydrogen fumarate group 7,6 0,27 3,9 0,08* 9,2 0,34
4,0 0,22*
GBB group 8,1 0,25 4,4 0,39 8,4 0,29
4,5 0,31
Each value represents the mean S.E.M. of 8-10 animals. *Significantly
different from
the control group (Tukey's test P< 0.05).
Insulin levels were decreased by 3-carboxy-N-ethyl-N,N-dimethylpropan-1-
aminium
hydrogen fumarate and 3-carboxy-N-ethyl-/V,/V-dimethylpropan-1-aminium
dihydrogen
phosphate (Table 2).
Table 2
Effects of test compounds at a dose of 10mg/kg on plasma insulin concentration
in
fasted ApoE-i- mice
Week 8 Week 16
Control group 1,39 0,13 1,26 0,05
3-carboxy-/Vethyl-N,Aklimethylpropan-1-
aminium dihydrogen phosphate group 0,60 0,06* 0,97 0,11*
3-carboxy-/Vethyl-N,Aklimethylpropan-1-
aminium hydrogen fumarate group 0,62 0,06* 0,85 0,06*
GBB group 1,30 0,11 1,20 0,10

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s
Each value represents the mean S.E.M. of 8-10 animals. *Significantly
different from
the control group (Tukey's test P< 0.05).
Results presented in Table 1 and Table 2 show that 3-carboxy-N-ethyl-N,N-
dimethylpropan-1-aminium hydrogen fumarate and 3-carboxy-N-ethyl-N,N-
dimethylpropan-1-aminium dihydrogen phosphate have a glucose and insulin level
lowering properties, thus demonstrating efficacy in prevention and treatment
of
diabetes.