WO2007003271A1

WO2007003271A1 - Salt of embusartan

Info

Publication number: WO2007003271A1
Application number: PCT/EP2006/005934
Authority: WO
Inventors: Hans-Georg Lerchen; Alfons Grunenberg; Jörg Keldenich; Ursula Krenz; Nicole Diedrichs; Andreas Knorr; Axel Kretschmer
Original assignee: Bayer Healthcare Ag
Priority date: 2005-06-30
Filing date: 2006-06-21
Publication date: 2007-01-11
Also published as: DE102005030522A1

Abstract

The invention relates to the diethanolamine salt of embusartan, methods for the production thereof, medicaments containing the same, and the use thereof for treating diseases.

Description

Salt of the Embusartan

The present invention relates to the diethanolamine salt of embusartan, to a process for its preparation, to medicaments containing it and to its use for the treatment of diseases.

Renin, a proteolytic enzyme, is known to cleave the decapeptide angiotensin I from the angiotensinogen in vivo, which in turn degrades in the lungs, kidneys or other tissues to the hypertensive octapeptide angiotensin II. The various effects of angiotensin II, such as vasoconstriction, Na ⁺ retention in the kidney, aldosterone release in the adrenal gland and increased tone of the sympathetic nervous system act synergistically in the sense of increasing blood pressure.

In addition, angiotensin II has the property of promoting the growth and proliferation of cells such as cardiomyocytes and smooth muscle cells, and these proliferate and proliferate in various disease states (e.g., hypertension, atherosclerosis, and heart failure).

One possible approach to engaging in the renin-angiotensin (RAS) system, in addition to inhibiting renin activity, is the inhibition of angiotensin converting enzyme (ACE) activity and the blockade of angiotensin II receptors.

Embusartane belongs to the structural class of the pyridone-substituted biphenyls disclosed in EP 0 542 059. Embusartane, which is methyl 6-butyl-1- {[3-fluoro-2 '- (1H-tetrazol-5-yl) biphenyl-4-yl] methyl} -2-oxo-1,2-dihydropyridine-4-carboxylate its preparation, specific angiotensin II antagonist activity and medical use is described in EP 0 594 019.

The object of the present invention is to provide a form of embusartan having an improved physicochemical profile, in particular solubility, while maintaining sufficient stability.

The acidic tetrazole residue of the embusartan lends itself to salt formation with bases, for example amine bases. However, the methyl ester group of the embusartan is easily cleaved under basic conditions. Various salts of embusartan with bases show cleavage of the methyl ester and / or do not form sufficiently stable crystals. The present invention is the diethanolamine salt of Embusartan 2-hydroxy-N- (2-hydroxyethyl) ethanaminium 5- (4 '- {[6-butyl-4- (methoxycarbonyl) -2-oxopyridin-1 (2H) -yl ] - methyl} -3'-fluorobiphenyl-2-yl) tetrazole-l-id and corresponds to the compound of formula (I)

Another object of the present invention is the compound of formula (I) in a crystalline form having a melting point of 136 ° C.

Surprisingly, the base-labile methyl ester group of the compound of formula (I) is not cleaved. The compound of formula (I) is characterized by a high solubility and stability.

The present invention furthermore relates to a process for the preparation of the compound of the formula (I). For this purpose, embusartane is mixed with diethanolamine, preferably 1 mol equivalent, based on the molar amount of embusartane used, in an inert solvent, for example dioxane, and the mixture is homogenized until a clear solution is formed. The mixture is optionally concentrated under reduced pressure. The mixture can also be lyophilized under reduced pressure. The isolated product is usually amorphous.

To prepare the crystalline form, the amorphous compound of the formula (I) is stirred in an inert solvent, preferably a mixture of n-butanol and water, for several days, preferably up to 7 days, the crystals are isolated and optionally dried. Figure 2 shows the X-ray diffraction of the crystalline form of the compound of formula (I).

The compound of the formula (I) can be suitably applied, e.g. oral, parenteral, pulmonary, nasal, sublingual, lingual, buccal, rectal, dermal, transdermal, conjunctival, otic or as an implant or stent.

For these routes of administration, the compound of the formula (I) can be administered in suitable administration forms. For oral administration are according to the prior art functioning rapidly and / or modified the compound of formula (I) donating application forms, such as tablets (uncoated or coated tablets, for example with enteric or delayed dissolving or insoluble coatings, the release the compound according to the invention), rapidly disintegrating tablets or films / wafers, films / lyophilisates, capsules (for example hard or soft gelatine capsules), dragees, granules, pellets, powders, suspensions or aerosols in the oral cavity.

Parenteral administration can be accomplished by bypassing a resorption step (e.g., intravenously, intraarterially, intracardially, intraspinal, or intralumbar) or by resorting to absorption (e.g., intramuscularly, subcutaneously, intracutaneously, percutaneously, or intraperitoneally). For parenteral administration are suitable as application forms u.a. Injection and infusion preparations in the form of suspensions, lyophilisates or sterile powders.

For the other routes of administration are suitable, for example Inhalation medicaments (including powder inhalers, nebulizers), lingual, sublingual or buccal tablets, films / wafers or capsules, suppositories, ear or ophthalmic preparations, vaginal capsules, aqueous suspensions (lotions, shaking mixtures), lipophilic suspensions, ointments, creams, transdermal therapeutic systems (such as patches), pastes, scattering powders, implants or stents.

The compounds according to the invention can be converted into the stated administration forms. This can be done in a conventional manner by mixing with inert, non-toxic, pharmaceutically suitable excipients. These adjuvants include, among others. Excipients (for example microcrystalline cellulose, lactose, mannitol), solvents (for example liquid polyethylene glycols), emulsifiers and dispersants or wetting agents (for example sodium dodecyl sulfate, polyoxysorbitol oleate), binders (for example polyvinylpyrrolidone), synthetic and natural polymers (for example albumin), stabilizers ( For example, antioxidants such as ascorbic acid), dyes (eg, inorganic pigments such as iron oxides) and flavor and / or odoriferous.

Another object of the present invention are pharmaceutical compositions containing at least the compound of formula (I), usually together with one or more inert, non-toxic, pharmaceutically suitable excipients such as binders, fillers, etc., as well as their use for the purposes mentioned below.

The compound of formula (I) has a specific angiotensin E antagonist effect, as it competitively inhibits the binding of angiotensin II to the receptors. She suppresses the Vasoconstrictor and aldosterone secretion stimulating effects of angiotensin H In addition, it inhibits the proliferation of smooth muscle cells.

The compound of the formula (I) can therefore be used in medicaments for the treatment and / or prophylaxis of arterial hypertension and atherosclerosis. In addition, it can be used for the treatment and / or prophylaxis of coronary heart disease, heart failure, brain disorders, ischemic brain disorders, peripheral circulatory disorders, kidney and adrenal function disorders, bronchospastic and vascular diseases of the respiratory tract, sodium retention and edema.

Because the compound of formula (I) surprisingly exhibits PPAR γ activity, the compound of formula (I) can therefore be used in blood sugar control drugs for the treatment and / or prophylaxis of diabetes mellitus or for the treatment of patients with abnormal PPAR γ function.

Due to PPAR γ activity, the compound of formula (I) can be used not only to control blood sugar levels in patients with diabetes mellitus or in pre-diabetic patients, but also to improve lipid metabolism or to treat arteriosclerosis.

Similarly, patients with metabolic syndrome, Syndrome X, impaired glucose tolerance (IGT), insulin resistance, or impaired fasting glucose (IFG). These patients may have elevated blood sugar levels of 110 mg / dL or higher. Examples of abnormal PPAR γ activity may be proliferative, autoimmune, immunomodulatory or inflammatory diseases.

embodiments

NMR, LC-MS and HPLC methods:

Method 1 (standard conditions for analytical HPLC):

Column: XTerra 3.9 x 150 WAT 186000478; Temperature: RT; Flow: 1 ml / min;

Eluent A: 10 ml 70% perchloric acid in 2.5 l water, eluent B: acetonitrile, gradient: 0.0 min 20% B → 1 min 20% B → 4 min 90% B → 9 min 90% B → 10 min 20% B. Method 2 (standard conditions for LC-MS, MHZ-ZI-HYD-I):

Device type MS: Micromass ZQ; Device type HPLC: Waters Alliance 2795; Column: Phenomenex Synergi 2μ Hydro-RP Mercury 20mm x 4mm; Eluent A: 1 l of water + 0.5 ml of 50% formic acid, eluent B: 1 l of acetonitrile + 0.5 ml of 50% formic acid; Gradient: 0.0 min 90% A -> 2.5 min 30% A -> 3.0 min 5% A -> 4.5 min 5% A; Flow: 0.0 min 1 ml / min, 2.5 min / 3.0 min / 4.5 min 2 ml / min; Oven: 50 ^° C .; UV detection: 210 nm.

NMR:

NMR measurements were performed on a Bruker DMX500 with a proton frequency of 500.13 MHz. Samples were dissolved in DMSO-d ₆ and measured at a temperature of 302K.

Example 1:

2-hydroxy-N- (2-hydroxyethyl) ethanaminium 5- (4 '- {[6-butyl-4- (methoxycarbonyl) -2-oxo-pyridin-1 (2H) -yl] methyl} -3'-fluorobiphenyl 2-yl) tetrazol-l-id

a) amorphous form:

5 g (10.83 mmol) of methyl 6-butyl-1- {[3-fluoro-2 '- (1H-tetrazol-5-yl) biphenyl-4-yl] methyl} -2-oxo-1, 2 dihydropyridine-4-carboxylate (Embusartan) are taken up in 50 ml of dioxane and 5 ml of methanol and treated with 1.14 g (10.83 mmol) of diethanolamine. After 2 min treatment in an ultrasonic bath, a clear solution is formed. The mixture is concentrated under high vacuum at room temperature, whereby the dioxane freezes. The contents of the flask are then lyophilized. The lyophilisate is again taken up in dioxane and lyophilized again. The salt is obtained in quantitative yield.

HPLC (method 1): R ₁ = 5.28 min; LC-MS (Method 2): R ₁ = 2.21 min; m / z = 462 (M + H) ⁺

¹ H-NMR (500 MHz, DMSO-Cl ₆ ): δ = 0.88 (t, 3H), 1.32 (m, 2H), 1.52 (m, 2H), 2.66 (t, 2H), 3.0 (t, 4H), 3.63 (t, 4H), 3.88 (s, 3H), 5.3 (s, 2H), 6.53 (s, IH), 6.66 (t, IH), 6.84 (s, IH), 6.89 (d, IH), 6.93 (d, IH), 7.32 (m, IH), 7.4 (m, 2H), 7.6 (m, IH).

b) crystalline form:

6.1 g (10.9 mmol) of the salt prepared under a) are stirred in 3 ml of a mixture of 85% n-butanol and 15% water at RT. After 7 days, it is filtered with suction through a frit and Ih sucked dry with air. It is dried for a further 5 days in the air. 3.94 g (64%) of a crystalline substance are obtained.

TLC: acetonitrile / water (20/1); R _f = 0.26

HPLC (Method 1): R, = 5.28 min;

LC-MS (Method 2): R, = 2.21 min; m / z = 462 (M + H) ⁺

The NMR spectrum is identical to the NMR spectrum measured before stirring.

Crystallinity and hygroscopicity:

The high enthalpy of fusion (108 J / g) of Example 1 b) determined with a differential scanning calorimeter (DSC, Fig. 1) and the X-ray diffraction pattern (Fig. 2) show that Example 1 b) is present as a crystalline salt , The melting behavior (DSC melting point 136 ° C, Fig. 1, red curve) shows _? that the investigated salt has a high purity. Example 1 b) is not hygroscopic. The TGA thermogram (Thermogravimetric Analysis, Fig. 1, black curve) shows no mass loss between ambient temperature and 100 ⁰ C. During storage at high relative humidity (85% rh at 25 ° C, one week) takes the TGA mass loss for only 0.1% too.

The DSC and TGA thermograms were taken with a Perkin Elmer DSC 7 and TGA 7 analysis system. The X-ray diffractogram was recorded using a Stoe transmission diffractometer.

stability studies

pH stability:

0.3 mg of Example 1 b) are weighed into a 2 ml HPLC vial and treated with 0.5 ml of acetonitrile. To dissolve the substance, the sample vial is placed in an ultrasonic bath for approx. 10 seconds dipped. Subsequently, 0.5 ml of the respective buffer solution is added and the sample is again treated in an ultrasonic bath.

Used buffer solutions:

pH 2: 0.03 M citric acid; 0.061 M sodium chloride; 0.0082 M hydrochloric acid

pH 4: 0.056 M citric acid; 0.044 M sodium chloride; 0.068 M sodium hydroxide solution

pH 6: 0.06 M citric acid; 0.160 M sodium hydroxide solution

pH 6.5: (6.186 g sodium chloride + 3.954 g sodium dihydrogen phosphate) / 1 adjusted to pH 6.5 with 1N NaOH

pH 7.5: (1.23 g citric acid anhydrous + 22.65 g disodium hydrogen phosphate) / 1

pH 8: 0.013 M borax; 0.021 M hydrochloric acid

pH 9: 0.013 M borax; 0.0046 M hydrochloric acid

Over a period of 24 h at 37 ° C., in each case 5 .mu.l of the sample solution are analyzed by HPLC for their content of active ingredient per hour. Quantification is made over the area percentages of the peaks.

HPLC method: Agilent 1100 with DAD (Gl 314A), binary pump (Gl 312A), autosampler (Gl 329A), column oven (G1316A); Thermostat (G1330A)

Column: Kromasil TUO Cl 8 60 x 2.1 3.5μ;

Column temperature: 37 ° C;

Eluent A: water + 5 ml perchloric acid / l

Eluent B: acetonitrile;

Gradient: 0-1.0 min 98% A, 2% B; 1, 0-9.0 min 2%, A 98% B; 9.0-13.0 min 2% A, 98% B; 13.0-13.5 min 98% A, 2% B; 13.5-15.0 98% A, 2% B.

Flow rate: 0.75 ml / min;

Detection: 210 nm.

Results: The stability of Example 1 b) is characterized as follows:

Buffer pH content in% after 24 h at 37 ⁰ C 2.0 100.0 after 24 h at 37 ⁰ C 4.0 100.0 after 24 h at 37 ⁰ C 6.0 100.0 after 24 h at 37 ⁰ C 6.5 100.0 after 24 h at 37 ^° C. 7.5 97.0 after 24 h at 37 ^° C. 8.0 83.0 after 24 h at 37 ^° C. 9.0 4.0

Thermal stability:

10 mg of Example 1 b) are transferred to a 10 ml amber glass bottle and sealed tight with a rubber stopper including crimp cap. The bottle is stored in the drying oven of WTB binder (VD23) at 9O ⁰ C for one week. After this time, 0.3 mg are removed and dissolved in 1.0 ml of water / acetonitrile 1: 1. This sample is analyzed by HPLC under the conditions described above.

The chromatogram of the test compound at time t ₀ before storage serves as comparison.

Results:

For Example 1 b) results after storage of 7 days at 90 ⁰ C, a content of 93.6% active ingredient based on 3Sn starting value of 100%.

In comparison, results for the known Embusartan potassium salt, described in EP 0 594 019, after storage for 7 days at 9O ⁰ C, a content of 33% of active ingredient based on the initial value of 97%.

Determination of solubility:

The test compound is dissolved in water or aqueous buffer (specified in the solubility results). This solution is shaken for 24 h at room temperature. After ultra-centrifugation at 224 000 g for 30 min. The supernatant is diluted with DMSO and analyzed by HPLC. Quantification is via a two-point calibration curve of the test compound in DMSO. The solubility is expressed in mg / l. HPLC method: Agilent 1 100 with DAD (G1315A), quat. Pump (G1311A), autosampler CTC HTS PAL, degasser (G1322A) and column thermostat (G1316A); Column: Zorbax Extend-C18 3.5μ; Temperature: 40 ° C; Eluent A: water / phosphoric acid pH2; Eluent B: acetonitrile; Flow rate: 0.7 mL / min; Gradient: 0-0.5 min 85% A, 15% B; Ramp: 0.5-3 min 10%, A 90% B; 3-3.5 min 10% A, 90% B; Ramp: 3.5-4 min 85% A, 15% B; 4-5 minutes 85% A, 15% B.

Results:

Example 1 b) is characterized for solubility as follows:

pH SOL [mg / 1]

6.6 420

Degradation is not observed.

In comparison, embusartan for solubility is characterized as follows:

pH SOL [mg / 1]

2 4

4 1 1

6 240

Above pH 6, significant degradation of the methyl ester to the carboxylic acid is observed.

In further comparison, the Embusartan potassium salt for solubility is characterized as follows:

pH SOL [mg / l]

2.0 4.6

Above pH 2, degradation of the methyl ester to the carboxylic acid is already observed.

method

A) Radiotelemetric blood pressure measurement on awake rats

A commercially available telemetry system from DATA SCIENCES INTERNATIONAL DSI, USA is used for the blood pressure measurement on awake rats described below.

The system consists of 3 main components:

1. Implantable transmitter 2. receivers, which have a multiplexer with a

3. Data acquisition computer are connected.

The telemetry system allows a continuous recording of blood pressure heart rate and body movement on awake animals in their habitual habitat.

animal material

The tests are carried out on adult female rats weighing> 200 g. The experimental animals are kept individually in Makrolon cages type 3 after transmitter implantation. You have free access to standard food and water.

The day - night rhythm in the experimental laboratory is changed by room lighting at 6:00 in the morning and at 19:00 in the evening.

transmitter implantation

The rat telemetry transmitters (TAM PA - C40, DS) are surgically implanted into the experimental animals under aseptic conditions at least 14 days before the first trial. The animals so instrumented are repeatedly used after healing of the wound and implant ing.

For implantation, the fasting animals are anesthetized with pentobabital (Nembutal, Sanofi: 50 mg / kg i.p.) and shaved and disinfected on the ventral side. After opening the abdominal cavity along the alba line, the liquid-filled measuring catheter of the system above the bifurcation is inserted cranially into the descending aorta and secured with tissue adhesive (VetBonD ™, 3M). The transmitter housing is fixed intraperitoneally to the abdominal wall musculature and the wound is closed in layers.

Postoperatively, an antibiotic is administered for infection prevention (Tardomyocel COMP Bayer 1ml / kg s.c.)

Substances and solutions

Unless otherwise described, the substances to be tested are each administered orally to a group of animals (n = 6) by gavage. According to an application volume of 5 ml / kg body weight, the test substances are dissolved in suitable solvent mixtures or suspended in 0.5% Tylose.

A solvent-treated group of animals is used as a control. experimental procedure

The existing telemetry measuring device is configured for 24 animals. Each trial is registered under a trial number (VYear month day).

The instrumented rats living in the plant each have their own receiving antenna (1010 receivers, DSI).

The implanted transmitters can be activated externally via a built-in magnetic switch. They will be put on the air during the trial run. The emitted signals can be recorded online by a data acquisition system (Dataquest ™ A.R.T. for Windows, DSI) and processed accordingly. The storage of the data takes place in each case in a folder opened for this purpose which carries the test number.

In the standard procedure duration is measured over every 10 seconds

• Systolic blood pressure (SBP)

Diastolic blood pressure (DBP)

• Mean arterial pressure (MAP)

• heart rate (HR)

• Activity (ACT)

The measured value acquisition is repeated computer-controlled in 5-minute intervals. The source data collected as absolute values are corrected in the diagram with the currently measured barometric pressure and stored in individual data. Further technical details can be found in the extensive documentation of the manufacturer (DSI).

Unless otherwise stated, the administration of the test substances will take place at 9 o'clock on the day of the experiment. Following the application, the parameters described above are measured for 24 hours.

evaluation

After the end of the test, the collected individual data are sorted with the analysis software (DATAQUEST TM ART TM ANALYSIS). The blank value is assumed to be 2 hours before the application, so that the selected data record covers the period from 7:00 am on the test day to 9:00 am on the following day. The data is smoothed over a presettable time by averaging (15 minutes average, 30 minutes average) and transferred as a text file to a disk. The presorted and compressed measured values are transferred to Excel templates and displayed in tabular form. The filing of the collected data takes place per experiment day in a separate folder that bears the test number. Results and test reports are sorted in folders and sorted by paper.

Result

After administration of 30 mg / kg p.o. the compound of formula (I) the maximum effect is increased by 50% compared to Embusartan (30 mg / kg p.o.). The duration of action tends to be prolonged, but the reflex tachycardia is not increased.

B) Cellular Transactivation Assay:

Principle of the test: A cellular assay is used to identify activators of the peroxisome proliferator-activated receptor gamma (PPAR-gamma).

Because mammalian cells contain various endogenous nuclear receptors that could complicate unambiguous interpretation of the results, an established chimera system is used in which the ligand-binding domain of the human PPARγ receptor is fused to the DNA binding domain of the yeast transcription factor GAL4. The resulting GAL4-PPARγ chimera is co-transfected into CHO cells with a reporter construct and stably expressed.

cloning:

The GALΦPPARγ expression construct contains the ligand binding domain of PPARγ (amino acids 203-506), which is PCR amplified and cloned into the vector pcDNA3.1. This vector already contains the GAL4 DNA binding domain

(Amino acids 1-147) of the vector pFC2-dbd (Stratagene). The reporter construct, which is five

Copies of the GAL4 binding site, upstream of a thymidine kinase promoter, results in

Expression of firefly luciferase (Photinus pyralis) after activation and binding of GAL4-PPARγ. Transactivation Assav: luciferase reporter

CHO (Chinese hamster ovary) cells are grown in DMEM / F12 medium (BioWhittaker) supplemented with 10% fetal calf serum, 1% penicillin / streptomycin (GIBCO), with a cell density of 2 x 10 ³ cells per well in a 384 Well plate (Greiner) sown. After culturing for 48 h at 37 ° C, the cells are stimulated. For this purpose, the substances to be tested are taken up in CHO-A-SFM medium (GIBCO) supplemented with 2.5% fetal calf serum, 1% penicillin / streptomycin (GIBCO) and added to the cells. After a stimulation time of 24 hours, luciferase activity is measured using a video camera. The measured relative light units result in a sigmoidal stimulation curve as a function of the substance concentration. The ECso values are calculated using the computer program GraphPad PRISM (version 3.02).

Result:

The compound of formula (I) shows an EC ₅₀ of 17 μM.

Claims

claims:

1. 2-Hydroxy-N- (2-hydroxyethyl) ethanaminium 5- (4 '- {[6-butyl-4- (methoxycarbonyl) -2-oxopyridin-1 (2H) -yl] methyl} -3' -fluorobiphenyl-2-yl) tetrazole-l-id corresponding to the compound of the formula (I)

2. A process for the preparation of the compound of formula (I), characterized in that Embusartan is reacted with diethanolamine in an inert solvent.

3. Medicaments containing at least the compound of formula (I) one or more inert, non-toxic, pharmaceutically suitable excipients.

4. Medicament according to claim 3 for the treatment and / or prophylaxis of arterial

Hypertension, atherosclerosis, coronary heart disease, heart failure, brain disorders, ischemic brain disorders, peripheral circulatory disorders, kidney and adrenal function disorders, bronchospastic and vascular diseases of the respiratory tract, sodium retention and edema.

5. Use of the compound of formula (I) for the manufacture of a medicament for the treatment and / or prophylaxis of arterial hypertension, atherosclerosis, coronary heart disease, heart failure, disorders of brain function, ischemic brain disorders, peripheral circulatory disorders, kidney and adrenal function, bronchospastic and vascular conditioned respiratory diseases, sodium retention and edema.

6. A method for the treatment and / or prophylaxis of arterial hypertension, atherosclerosis, coronary heart disease, heart failure, disorders of brain function, ischemic brain disorders, peripheral circulatory disorders, kidney and adrenal function disorders, bronchospastic and vascular diseases of the respiratory tract, Sodium retention and edema by administration of a pharmaceutically effective amount of the compound of formula (I).

7. Use of the compound of formula (I) for the manufacture of a medicament for the control of the blood sugar level, for the treatment and / or prophylaxis of diabetes mellitus or for the treatment of patients with abnormal PPAR γ function.