Patent Application
20100069450
The present invention relates to fumarate salt of the compound of formula:
in all its stereochemical configurations, a process for its preparation and its use in treating mood disorders, disorders of anxiety, depression and convulsive conditions, in the improvement of learning ability, in the reversal of amnesia, in resolving the abstinence syndrome from drugs and drugs of abuse.
More specifically, the invention concerns fumarate salt of rel-(3R,3aS,7aS)-3-benzyl-2-methyl-2,3,3a,4,5,6,7,7a-octahydrobenzo[d]isoxazol-4-one.
The compound rel-(3R,3aS,7aS)-3-benzyl-2-methyl-2,3,3a,4,5,6,7,7a-octahydrobenzo[d]isoxazol-4-one, also known as BTG 1640, is described in the International application with publication number WO93/17004 and belongs to a family of new psychoactive compounds.
Specifically, in the patent application with publication number WO93/17004 the tranquilizing activity of the whole compound family is proposed and particularly, the activity of the compound BTG 1640 and the effects thereof in learning ability and in the reversal of amnesia are tested.
According to such a document, the compound BTG 1640 is prepared as a yellow oil and, in tests demonstrating the proposed activity, it is used after dilution in PEG and distilled water.
In pharmaceutical practises, it is known that oil substances have a lot of drawbacks mostly linked to the difficult handling and formulation. The oils are in fact hard to weight, basically less stable to temperature variations, less soluble in ordinary solvents and therefore technically harder to dose for the preparation of pharmaceutical formulations.
It is generally advisable that the form of active ingredient is the crystalline solid one which normally shows better characteristics, specifically in terms of handling for activities of pharmaceutical formulation.
With regard to this, the patent application WO93/17004 cites the possibility of preparing the salt form of new psychoactive compounds of general Formula (I), by treating the free base of a compound of Formula (I) with the suitable free acid. Specifically, document WO93/17004 describes the hydrochloride of BTG 1640 obtained in the form of white crystalline powders.
By taking such a salt as a reference, and wishing therefore to avoid the use of free base BTG 1640 in oil form, in the step of drug formulation, the stability of such a hydrochloride salt as active principle was evaluated, in order to state the storage and preservation conditions of the drug. It is known that a pharmaceutical substance is deemed stable if it maintains the same initial characteristics, when subjected to different temperature and humidity conditions in time. As it will be demonstrated in the examples, the stability analysis showed that the hydrochloride salt of BTG 1640 is unstable at temperatures higher than 30° C., thereby refrigerator preservation at temperatures of 2-8° C. becomes advisable, in precautional way, not only for the single salt as active substance, but also for the different pharmaceutical formulations, which are nowadays developed and based on such an active principle.
Such a refrigerator preservation condition, although necessary, results rather disadvantageous. As a matter of fact, the observation of such preservation precautions is scarcely accepted by the patient, who results evidently not accommodated in the daily practise and in his/her activities, by having to keep the drug in the refrigerator, out of necessity.
It is still felt the need of providing the compound 3-benzyl-2-methyl-2,3,3a,4,5,6,7,7a-octahydrobenzo[d]isoxazol-4-one in a form which does not require peculiar preservation conditions.
An object of the present invention is therefore to provide a form of 3-benzyl-2-methyl-2,3,3a,4,5,6,7,7a-octahydrobenzo[d]soxazol-4-one which does not require peculiar preservation and storage conditions.
It is a further object to provide a salt of BTG 1640 which shows bioavailability characteristics at least comparable to those of hydrochloride salt of known BTG 1640.
The inventors of the present invention carried out studies and researches and as a result they surprisingly found out that the fumarate salt of:
shows a highly improved stability so as not to require peculiar preservation conditions, even after six months of preservation at 40° C./75% RH.
Therefore the invention concerns fumarate salt as recited in claim 1, a new process of preparation and its use as a medicament, specifically in treating mood disorders, disorders of anxiety, depression and convulsive conditions, in the improvement of learning ability, in the reversal of amnesia, in resolving the abstinence syndrome from drugs and drugs of abuse.
Preferably, according to the invention fumarate of rel-(3R,3aS,7aS)-3-benzyl-2-methyl-2,3,3a,4,5,6,7,7a-octahydrobenzo[d]isoxazol-4-one is provided, comprising the molecule in the two enantiomeric forms and as racemic mixture.
Advantages and further characteristics are indicated in the dependent claims.
In the present invention the molecule 3-benzyl-2-methyl-2,3,3a,4,5,6,7,7a-octahydrobenzo[d]isoxazol-4-one so as represented in formula I:
comprises in its definition any stereochemical configuration associated to chiral centers, as well as racemic mixtures and enantiomers obtainable through separation techniques known to skilled men and any mixture of two or more stereochemical compounds.
The invention will be now described in greater detail, particularly by making reference to the annexed figures wherein:
The invention therefore concerns the fumarate salt of molecule 3-benzyl-2-methyl-2,3,3a,4,5,6,7,7a-octahydrobenzo[d]isoxazol-4- one.
According to the invention, such salt can be obtained by treating the free base 3-benzyl-2-methyl-2,3,3a,4,5,6,7,7a-octahydrobenzo[d]isoxazol-4-one with fumaric acid, or alternatively, from hydrochloride salt of 3-benzyl-2-methyl-2,3,3a,4,5,6,7,7a-octahydrobenzo[d]isoxazol-4-one through treatment directed to free the molecule 3-benzyl-2-methyl-2,3,3a,4,5,6,7,7a-octahydrobenzo[d]isoxazol-4-one as free base and subsequent reaction with fumaric acid.
In another aspect, the invention concerns a process for the preparation of fumarate salt of 3-benzyl-2-methyl-2,3,3a,4,5,6,7,7a-octahydrobenzo[d]isoxazol-4-one according to claim 5, comprising the following steps:
Alternatively the free base 3-benzyl-2-methyl-2,3,3a,4,5,6,7,7a-octahydrobenzo[d]is oxazol-4-one of step i) can be obtained in a preceding step to step i), which provides for freeing said base from correspondent hydrochloride salt.
Specifically, the hydrochloride salt of 3-benzyl-2-methyl-2,3,3a,4,5,6,7,7a-octahydrobenzo[d]isoxazol-4-one can be subjected to subsequent extractions in dichloromethane to obtain the molecule 3-benzyl-2-metil-2,3,3a,4,5,6,7,7a-octahydrobenzo[d]isoxazol-4-one as free base in form of a transparent oil of slightly brown colour. To such a solution, fumaric acid can be added in presence of an ice bath at about 2-8° C. in form of crystals. Further preparation processes can be of course provided for by a synthesis organic technician. The salt so obtained can be furthermore optionally subjected to purification methods, if necessary.
The fumarate salt so obtained resulted to be stable in different preservation conditions, as it will be demonstrated in examples, thus demonstrating itself more stable than hydrochloride salt.
The salt fumarate of 3-benzyl-2-methyl-2,3,3a,4,5,6,7,7a-octahydrobenzo[d]isoxazol-4-one resulted, surprisingly, not only more stable than the known hydrochloride, but, as it will be widely demonstrated in the following, it resulted more bioavailable, that is it showed a kinetic absorption profile in vivo better than the hydrochloride salt.
Furthermore, the fumarate salt of 3-benzyl-2-methyl-2,3,3a,4,5,6,7,7a-octahydrobenzo[d]isoxazol-4-one resulted surprisingly strongly less toxic than hydrochloride salt as it will be shown below.
Fumarate salt of the compound of formula I having improved stability can be combined to suitable excipients for formulating pharmaceutical compositions according to the invention and is capable to act as pharmaceutical active substance in the treatment of mood disorders, disorders of anxiety, depression and convulsive conditions, in the improvement of learning ability, in the reversal of amnesia generated for example by Alzheimer disease or vascular dementia, in resolving the abstinence syndrome from drugs and drugs of abuse.
The daily dose required to reach the effect in the treatment of the indicated pathology varies with the subject, by depending by age, body weight and health general state, but it can be provided for a dosage suitable for the oral or topic administration in the range from 1 to 100 mg, once or more times a day, and a dosage suitable for parental administration in the range from 0.1 to 100 mg, once or more times a day.
The fumarate salt of compound of formula I will be added to a pharmaceutically acceptable carrier and, optionally, to other excipients in order to obtain pharmaceutical compositions to be parenterally, orally or topically administered. By the term “pharmaceutically acceptable carrier” it is meant to include solvents, supporting agents, diluents and the like, which are used as additives in order to provide a carrier suitable to the administration of the salt of the invention.
Compositions of the present invention suitable for oral administration will be conveniently in the form of discrete units such as tablets, capsules, cachets, powders, or granules, or still as suspensions in a liquid.
More preferably, the composition of the invention for the oral administration will be in form of tablets.
The tablet according to the invention comprises preferably an amount from 1 to 100 mg, preferably from 1 to 50 mg, of fumarate salt of a compound of formula I per tablet unit. Advantageously, the tablet comprises also suitable excipients, such as pre-gel starch, microcrystalline cellulose, sodium starch glycolate, talc, lactose, magnesium stearate, sucrose, stearic acid and mannitol.
Preferably, the tablet comprises from 1.9% to 41.4% by weight of fumarate of 3-benzyl-2-methyl-2,3,3a,4,5,6,7,7a-octahydrobenzo[d]isoxazol-4-one, more preferably from 2.2% to 36% by weight with respect of the total weight of the tablet.
The composition for oral administration preferably will comprise from 1 to 100 mg of fumarate salt of 3-benzyl-2-methyl-2,3,3a,4,5,6,7,7a-octahydrobenzo[d]isoxazol-4-one.
Compositions for the parenteral administration will comprise conveniently sterile aqueous preparations.
The compositions for parenteral administration preferably will comprise from 0.1 to 100 mg of fumarate salt of 3-benzyl-2-methyl-2,3,3a,4,5,6,7,7a-octahydrobenzo[d]isoxazol-4- one.
Compositions for topical administration will be conveniently in form of creams, oils, ointments, emulsions, gels, aqueous solutions, spray solutions and plasters.
The compositions for topical administration preferably will comprise from 1 to 100 mg of fumarate of 3-benzyl-2-methyl-2,3,3a,4,5,6,7,7a-octahydrobenzo[d]isoxazol-4-one.
The invention will now described in greater details in the following examples, by way of non-limiting example of the invention and relating to the preparation of fumarate of BTG 1640 and to the evaluation of the stability, absorption efficacy and toxicity of the hydrochloride salt of the prior art and of the fumarate salt of the invention.
A. Preparation of the Free Base
10 g of BTG 1640 hydrochloride were solubilized in 70 ml of sodium bicarbonate at 5%. The obtained solution was transferred into a separatory funnel of 250 ml and 50 ml of dichloromethane were added. After strong stirring for 30 seconds, the solution was left to rest until the separation of two layers. The phase containing dichloromethane was then transferred in an Erlenmeyer flask and further 20 ml of dichloromethane were added to the separatory funnel. Strong stirring of the solution contained in the separatory funnel was then again carried out for 30 seconds and the solution was left to rest until a new separation of the two layers. Once dichloromethane phase was separated, which was added to the Erlenmeyer flask, the extraction with further 20 ml of dichloromethane was repeated. Three aliquots of dichloromethane were then anhydrified with sodium solphate, filtered on paper filter and evaporated at rotavapor at a temperature below 35° C. The residue obtained as a slightly brown coloured oil corresponded to the free base of BTG 1640.
B. Preparation of Fumarate Salt
1 g of free base BTG 1640 obtained as indicated in part A was then solubilized in 20 ml of methanol. 10 ml of a methanolic solution comprising 473 mg of fumaric acid (PM=116.07) were then added dropwise. During the addition, the solution of BTG 1640 was kept submerged in an ice bath.
5 ml of hexane (alternatively 5 ml of ether were employed) were then added and the solution was subjected to concentration until a residue. The residue was then retaken with 36 ml of a mixture consisting of methanol/diisopropilic ether in a ratio of 1:2 and the so obtained solution was transferred into refrigerator at 2-8° C. for 48 hours. At the end of 48 hours, the formed crystals were isolated through filtration and finally dried into a stove under vacuum at 35° C. Mother liquor was again concentrated to a residue and the residue was then retaken with 36 ml of diisopropilic ether. The obtained solution was then again placed into the refrigerator at 2-8° C. for 48 hours. At the end of 48 hours, the formed crystals were isolated through filtration and finally dried into a stove under vacuum at 35° C. The overall yield of the process was 89%.
The so obtained crystals were subjected to characterization in order to demonstrate that they corresponded to fumarate of BTG 1640.
A sample of crystals was then subjected to elementary analysis and the obtained results corresponded to the values calculated on the basis of the proposed chemical formula.
The spectrum 1H-NMR, which confirmed the formation of fumarate of BTG 1640, was then obtained.
1H-NMR (200 MHz, DMSO-d6) δ: 1.65-2.40 (m, 6H) for H5 H6 e H7; 2.41 (s, 3H) for N—CH3; 2.70-2.92 (m, 2H) for benzylic CH2; 2.92-2.98 (m, 1H) for H3a; 3.35 (br, 1H) H3; 4.25 (br, 1H) for H7a; 6.63 (s, 1H) for —CH═CH— fumarate; 7.22-7.40 (m, 5H) for aromatic structure.
Samples of 100 mg each of BTG 1640 hydrochloride were subjected to the following preservation conditions:
a) 5° C.;
b) 25° C. and 60% RH;
c) 30° C. and 65% RH;
d) 40° C. and 70% RH.
The purities were then analyzed periodically, initially every three months, in order to underline possible variations.
The results in the following Table 1 were obtained:
The obtained results have been represented in
Contemporaneously, the impurity analysis in the conditions a)-d) of T/% RH were carried out.
The results in the following Table 2 were obtained:
The results were then represented in
Samples of 100 mg each of BTG 1640 fumarate prepared according to example 1C were subjected to the following preservation conditions:
a) 5° C.;
b) 25° C. and 60% RH;
c) 30° C. and 65% RH;
d) 40° C. and 75% RH.
Quarterly and in an overall period of nine months its purity was then analyzed in order to underline possible variations. The results in the following Table 3 were obtained:
The results were then represented in
As a confirmation of such evidence, a test for the evaluation of the impurity percentage of the sample subjected to the same conditions a)-d) of T/% RH was parallely carried out. The experimental values in Table 4 were obtained.
The obtained results were represented in
Therefore it is evident from the tests carried out the higher stability of the fumarate salt in compare to hydrochloride salt of BTG 1640.
As comparison other salts of BTG 1640 were prepared, and exactly:
The stabilities were then evaluated by subjecting samples of 100 mg of each salt to the following analogous preservation conditions:
a) 5° C.;
b) 25° C. and 60% RH;
c) 30° C. and 65% RH;
d) 40° C. and 70% RH.
Quarterly and in an overall period of nine months their purity was then analyzed in order to underline possible variations.
The obtained results, together with those obtained in Example 2 for hydrochloride, in Example 3 for fumarate in order to make easier the comparison, were in Table 5 and represented in
As it is seen from
The methanesulphonate salt begins to degrade already after three months of stability in the cited conditions b)-d), in amount of course higher as the conditions become severer, i.e. from a) to d). Maleate salt shows an acceptable profile in conditions a) and b), while it shows considerable degradation if subjected to conditions c) already at sixth month and degrades in unacceptable way in condition d). Succinate salt, at the end, shows degradation at all observation conditions a)-d), already from the third month and considerably as higher as severer are preservation conditions, i.e. from a) to d).
Consistently to these results, the impurity profile of salt samples subjected to such preservation conditions showed an unacceptable increase of the same as the salt itself degraded, as shown by patterns represented in
Surprisingly, therefore fumarate showed a different and improved stability with respect to known hydrochloride and other prepared acid addition salts, thus turning out to be the best ingredient for formulating pharmaceutical compositions, which does not require peculiar preservation conditions.
The present invention succeeded in solving the technical problem of obtaining a form of stable BTG 1640 through identification of the fumarate salt.
Absorption extent and rate of molecule BTG 1640 in its forms of fumarate and hydrochloride salts were evaluated.
With this intent, the two salts were dispersed in an aqueous suspension of arabic gum at 5% thus obtaining two formulations, each one orally administered through oesophageous gavage at dose of 10 mg/kg (expressed as free base of BTG 1640) to five rats Sprague Dawley.
Plasma concentration of molecule BTG 1640 was determined at different time points.
The results in Table 7 were obtained.
From the table 7, it is evident that fumarate, even if provides a not significantly higher concentration of free and unchanged active principle BTG 1640 with respect to hydrochloride after two minutes from its administration, it provides a much higher concentration of free base after 5 minutes with respect to hydrochloride.
All in all, the availability of the active principle in the circulation is more favourable after administration of fumarate salt thus it can be expected an higher pharmacological activity.
Acute oral toxicity of salts hydrochloride and fumarate of BTG-1640 was evaluated.
The two salts were dispersed in an aqueous suspension, thus obtaining two formulations, each one orally administered through oesophageous gavage at dose of 2000 mg/kg bw (expressed as free base of BTG 1640) to one of two groups comprising three female CD-1 mice per group.
The observation lasted 14 days.
In all the animals of each group a reduction of the spontaneous locomotory activity was recorded in the first minutes after the treatment and quickly disappeared. Lethargy, ataxia, aggressiveness and vocalization were observed in the mice treated with hydrochloride BTG 1640 and two of them died in the first day following the administration of the entire dose. The body weight remained quite stable over the entire period of observation in all the experimental groups. Necropsy revealed the absence of alterations, anyway kidneys were sampled from each animal, in order to assess the possible renal toxicity of the salts. The histopathological examination of the group administered with hydrochloride showed small foci of acute tubular necrosis in the cortex, balloon degeneration of the epithelium and dilatation of convoluted tubules. In the group administered with fumarate only a mild dilatation of convoluted tubules was observed. On the basis of the discussed results, the administration of 2000 mg/kg bw, in female mice, of fumarate BTG 1640 resulted strongly less toxic, being followed by quickly reversible neurological signs, while the same dose of hydrochloride BTG 1640 was lethal in two animals out of three.
Some formulative examples for the preparation of tablets having weight of about 120 mg of pharmaceutical composition according to the invention comprising fumarate salt of BTG 1640 and directed to oral administration are indicated in the following.
| Number | Date | Country | Kind |
|---|---|---|---|
| MI2006A002102 | Nov 2006 | IT | national |
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/IB07/03291 | 10/31/2007 | WO | 00 | 5/1/2009 |
