FR2546062A1 - Pharmaceutical compositions based on symmetrical disulpholanes - Google Patents

Abstract

Pharmaceutical compositions having an inhibitory action on erythrocyte formation or destruction due to a genetic modification of haemoglobin or to parasites, based on a compound of formula: in which R represents hydrogen and R' represents an NH group, or alternatively R and R', taken with the nitrogen and carbon atoms to which they are attached, form a 1H-imidazol-2-yl, 1-methyl-1H-imidazol-2-yl, 4,5-dihydro-1H-imidazol-2-yl, 4,5-dihydro-1-methyl-1H-imidazol-2-yl, 2-thiazolyl, 4,5-dihydro-2-thiazolyl, 1H-triazol-3-yl, 1-methyl-1H-triazol-2-yl or 2-pyrimidinyl radical.

Description

Pharmaceutical compositions based on metallic disulfolanes.

 The present invention relates to pharmaceutical compositions for the inhibition of the malformation or destruction of erythrocytes due to a genetic modification of hemoglobin or parasites and therefore useful in the treatment of sickle cell anemia and malaria.

 Sickle cell disease is known to be a genetic disorder that causes an abnormality in the structure of hemoglobin, the amino acid Glu-6 of the beta chain being replaced by the amino acid Val to give hemoglobin S which polymerizes.

Said polymerization, during the deoxygenation of the red blood cell, causes the sickling of the latter which becomes rigid, circulates poorly and becomes blocked in the small vessels. Individuals carrying two sickle cell genes are thus under the permanent threat of a fatal complication
Certain disulfides have been described in the literature as inhibiting sickling; in particular, cystamine has been indicated as particularly active ("Developments of Therapeutic Agents for Sigle Cell Disease", Inserm Symposium, 1979,
North Holland: Amsterdam, Editors J. Rosa, Y. Beuzard, J. Hercules; p. 139-153).

dans laquelle R représente hydro gène, R' représente un groupe N.H2 ou bien R et R', pris avec les atomes d'azote et de carbone auxquels ils sont attachés, forment un radical 1H-imidazol-2-yle, l-méthyl
H-imidazol-2-yle, 4,5-dihydro-1H-imidazol-2-yle, 4,5-dihydro-1 méthyl-lH-imidazol-2-yle, thiazol-2-yle, 4,5-dihydrothiazol-2-yle, 1H-1, 2,4-triazol-3-yle, @-méthyl-1H-1, 2, 4-triazol-3-yle ou pyrimidin-2-yle, sont très actifs dans l'inhibition de la polymérisation de l'hémoglobine S et de la falciformation des globules rouges. It has now been found that symmetrical disulfolanes of formula I

in which R represents hydrogen gene, R 'represents an N.H2 group or else R and R', taken with the nitrogen and carbon atoms to which they are attached, form a 1H-imidazol-2-yl radical, l- methyl
H-imidazol-2-yl, 4,5-dihydro-1H-imidazol-2-yle, 4,5-dihydro-1 methyl-1H-imidazol-2-yl, thiazol-2-yl, 4,5-dihydrothiazol -2-yle, 1H-1, 2,4-triazol-3-yle, @ -methyl-1H-1, 2, 4-triazol-3-yle or pyrimidin-2-yle, are very active in inhibition hemoglobin S polymerization and red blood cell sickling.

 It has also been found that the disulfolanes @ ymetrics of formula I above are active on parasites affecting drythrocytes, such as Plasmodia and Babesiae. More particularly, they show schizonticidal activity in vitro on "Plasmodium falciparum" and in vi @ o eur "Pla @ m @ dium berghei".

 Therefore, the expression "inhibition of the ma @ - formation or destruction of erythrocytes", as used here, defines an inhibition which can either be said to be hemoglobin or indirect by inhibition of the growth of parasites in the red blood cell.

 The inhibition of the falcifoxation of human erythrocytes was evaluated according to the method which consists in washing and incubating, in a saline buffer (pR 7.40 - 0.15 M), the red blood cells of sickle cell for one hour at 370C in a bath. blends with circular stirring in the presence of the above product at different concentrations. In this operation, the product / hemoglobin molar ratio is initially tested in the range from 0.5 to 20.

Depending on the results obtained, this report is modified. At the end of the incubation, the excess product is removed by washing. The cell suspension adjusted to a hematocrit of 5% is transferred to an Erlenmeyer flask and incubated at 370C under a stream of a humidified mixture of nitrogen and oxygen. The oxygen concentration is regulated by a gas mixture pump. The effluent gas is transferred to another tube containing formaldehyde. At the end of the incubation, the cell suspension is transferred to formaldehyde by simply inverting the flask.

The proportion of deformed cells and those with sickle cell characteristics (with two filiform extensions) is evaluated using a microscope having an interference optic of NOMARSKY. The sickling inhibition was calculated according to the following formula:
% sickle cells -% sickle cells in
witnesses presence of drugs
% sickle cells from control
Table I shows the percentage inhibition of erythrocyte sickling for certain compounds included in formula I above in comparison with cystamine.

TABLE I
Compound% inhibition cystamine 33% dithiodiformamidine 100% 2,2'-dithiodi-1H-imidazole 80% 2,2'-dithiodi (4,5-dihydrothiazole) 90% 3,3'-dithiodi-1H-1,2,4 -triazole 100% 2,2'-dithiodipyrimidine 50 7.

 From this table it appears that the compounds which are included in formula I above inhibit sickle cell sickle cell erythrocyte sickling and are much more active than the reference compound.

 Thus, the present invention relates to pharmaceutical compositions for the inhibition of the malformation or destruction of erythrocytes due to a genetic modification of hemoglobin or to parasites containing, as active ingredients, disulfolanes symmetrical of formula I above, as well as their possible pharmaceutically acceptable addition salts.

 In the pharmaceutical compositions of the present invention for oral, sublingual, subcutaneous, intramuscular, intravenous, transdermal or rectal administration, the active ingredients of formula I above can be administered in unit administration forms, as a mixture with conventional pharmaceutical carriers, animals and humans for the treatment of sickle cell disease, malaria or babeiosis.

Among the suitable unit forms of administration are the oral forms such as tablets, capsules, powders, granules and oral solutions or suspensions and the forms of sublingual and oral administration, as well as forms of subcutaneous, intramuscular or intravenous administration, possibly by infusion, and rectal.

 In order to obtain the desired effect, the dose of active principle can vary between 0.1 and 100 mg per kg of body weight per day.

 Each unit dose can contain from 10 to 1000 mg of active ingredient in combination with a pharmaceutical carrier.

This unit dose can be administered 1 to 4 times a day, possibly by infusion, for the treatment for example of sickle cell anemia, malaria and babébiosis.

 The powders are simply prepared for oral administration by grinding the active compound to a suitable fineness and mixing with a similarly ground diluent. The diluent can be an edible carbohydrate material such as starch. it is advantageous that a sweetening agent or a sugar, as well as a flavoring oil, be present.

 The granules intended for the reconstitution of an oral liquid preparation are prepared using water-soluble diluents, the active compound and a water-soluble diluent such as sucrose, glucose, etc. are wetted with a binder such as acacia mucilage, gelatin solution, methylcellulose solution and passed through a sieve, forcing to form granules which are dried. It is advantageous to introduce into the composition a suspending agent such as tragacanth.

 The capsules are made by preparing a powder mixture as described above and putting it in shaped gelatin sheaths. As an adjunct to the filling operation, it is advantageous to add a lubricant such as talc, magnesium stearate and calcium stearate to the powder mixture before the filling operation.

 The tablets are made by preparing a powder mixture, granulating or cutting, adding a lubricant and compressing to form tablets. The pulverulent mixture is prepared by mixing the desired compound, suitably ground, with a diluent or a base such as starch, sucrose, kaolin, bicalcium phosphate, etc. The pulverulent mixture can be granulated by wetting with the binder such as syrup, starch paste or acacia mucilage and by forcing through a sieve. As a method other than granulation, the pulverulent mixture can be cut up, that is to say, pass it through the tablet machine and break the resulting tablets imperfectly formed into fragments (sections).

You can lubricate the sections to prevent them from sticking to the tablets by forming cubes, adding a stearate salt, talc or mineral oil. The lubricant mixture is then compressed to form tablets.

 The tablets can advantageously be provided with a protective coating consisting of a gummelac sealer coating, a coating of sugar and methylcellulose, and a varnish coating of carnauba wax, or else they can be treated in another way. so that they have a prolonged or delayed activity and that they continuously release a predetermined amount of active ingredient.

Fluids for oral administration are prepared in unit dosage forms such as syrups where each teaspoon of composition contains a predetermined amount of the active compound to be administered
A preparation in the form of a syrup or elixir may contain the active ingredient together with a sweetener, methylparaben and propylparaben as antiseptics as well as a flavoring agent and an appropriate color
For rectal application, suppositories are used which are prepared with binders which melt at rectal temperature, for example cocoa butter or polydthylene glycols.

 For parenteral application, aqueous suspensions, isotonic saline solutions or sterile injectable solutions are used which contain pharmacologically compatible dispersing and / or wetting agents, for example propylene glycol or butylene glycol.

 The active principle can also be formulated in the form of microcapsules, optionally with one or more carriers or additives.

 The compounds of formula I above are known in the literature or they can be easily prepared by oxidation of the corresponding thiols according to well-known synthetic methods (Houben-Weil, Methoden der Organiachen Chimie, Vol. IX, p. 19, 4th Ed ., G. Thieme-Verlag, Stuttgart).

 The following examples illustrate the invention without, however, limiting it.

Preparation
The compound of formula I above where R represents hydrogen and R 'represents an NH2 group (dithiodiformamidine) is a well known product.

 The other disulfolanes of formula I are also described in the literature and are prepared by oxidation of the corresponding thiols with iodine, with bromine or in air. By way of example, the preparation of 2,2'-dithiodiimidazole is given below.

 In a flask fitted with a condenser, mercaptoimidazole 1 g is placed in a mixture of dichloromethane (10 ml) and 5% sodium bicarbonate (20 ml). The flask is immersed in a water bath and a bromine solution (0.26 ml) is added very slowly to the well-stirred mixture. During the addition the yellow color of the bromine is disparate. The solution is left stirring for another hour after all the bromine has been added.

The solution is filtered and an insoluble product is recovered. 230 mg of 2,2'-dithiodiimidazole are thus obtained.

Yield: 23.2 7; F. 158 C.

Example 1
Tablets having the following composition: 2,2'-dithiodi-1H-imidazole 500 mg glycine 120 mg microgranular cellulose 70 mg precipitated silica 18 mg carboxymethyl starch 30 mg magnesium stearate ll mg talc 11 mg
The calculated amounts of the components are mixed for 30 minutes, then granulated to dryness and passed through the mixture through a 1.6 mm mesh screen. It is then compressed using a punch having the shape of a small stick. This gives tablets each weighing 760 ing and each containing 500 mg of active ingredient.

Example 2
Tablets are prepared according to Example 1. The tablets thus obtained are coated with a suspension of dibutyl phthalate, polymethyl butyl dimethylaminoethyl, polyethylene glycol 1500, precipitated silica, titanium dioxide and talc in a 1: 1 acetone: isopropanol mixture having a dry residue concentration of approximately 10%. Coated tablets are thus obtained each weighing 780 mg and each containing 500 mg of active principle.

Example 3
A granule is prepared intended for the reconstitution of an oral liquid preparation having the following composition 2,2'-dithiodi (4,5-dihydrothiazole) 3.60 g sucrose 50.00 g carboxymethyl thylcellulose 0.80 g citric acid 0 , 10 g trisodium citrate 0.90 g sodium benzoate 0.25 g sodium saccharin 0.15 g flavoring 0.50 g
The volume of granule thus obtained is brought to 100 ml with water for syrups. A unit dose of 5 ml of the extemporaneous syrup thus obtained contains 180 mg of active principle.

 To prepare the granule, the calculated quantities of all the components except the sucrose are sprayed, then the powder thus obtained is mixed with the sucrose until a homogeneous granule is obtained.

Example 4
A granule is prepared according to Example 3 intended for the reconstitution of an oral liquid preparation having the following composition 3.3'-dithiodi-1H-1,2,4-triazole 7.00 g sucrose 46.60 g sodium carboxymethylcellulose 0.90 g citric acid 0.10 g trisodium citrate 0.90 g sodium benzoate 0.25 g sodium saccharin 0.15 g flavoring 0.50 g
A unit dose of 5 ml of the extemporaneous syrup thus obtained contains 350 mg of active principle.

Example 5
A granule is prepared according to Example 3 for the reconstitution of an oral liquid preparation having the following composition 2,2'-dithiodipyrimidine 8.00 g sucrose 45.60 g sodium carboxymethylcellulose 1.00 g citric acid 0.10 g citrate trisodium 0.90 g sodium benzoate 0.25 g sodium saccharin 0.15 g flavoring. 0.50 g
A unit dose of 5 ml of the extemporaneous syrup thus obtained contains 400 mg of active principle.

Example 6
Tablets having the following composition are prepared dithiodi formamidine 350 mg microcrystalline cellulose 100 mg lactose 125 mg magnesium stearate 10 mg talc 15 mg
The powders are passed through a 0.3 mm sieve, then the ingredients are mixed until a homogeneous mixture is obtained which is compressed and granulated. The granules thus obtained are used to form tablets by compression.

Weight of one tablet: 600 mg.

Example 7
Tablets having the following composition 2,2'-dithiodi (1-methyl-1H-imidazole) 150 mg microcrystalline cellulose 75 mg talc 15 mg polyvinylpyrrolidone 30 mg precipitated silica 25 mg magnesium stearate 5 mg
All the ingredients, except the lubricant, are intimately mixed in a mixer-kneader for 15 minutes, then kneaded by gradual addition of water. The mass is passed through a 1.25 mm sieve. The granulate is dried in an oven with forced ventilation until a relatively low residual humidity (approximately 2%) is obtained. The granule is made uniform, the lubricant is added and tablets are formed by compression.

Weight of one tablet: 300 mg.

 In the same way, tablets containing 250 mg of active principle are prepared.

Example 8
Tablets are prepared having the following composition: 2,2'-dithiodi (4,5-dihydrothiazole) 150 mg carboxylated thyl starch 10 mg microcrystalline cellulose 85 mg lactose 135 mg hydrogenated castor oil 10 mg magnesium stearate 5 mg operating as described in Example 8. The tablets thus obtained are covered with a coating having the following composition butyl phthalate 0.300 mg dimethylaminoethyl butyl methacrylate 1.850 mg polyethylene glycol 1500 0.080 mg precipitated silica 0.020 mg talc 0.900 mg titanium dioxide 1.850 mg dissolved in a solvent which is eliminated by evaporation in an oven with forced ventilation. Weight of one tablet: 400 mg.

Example 9
Suppositories having the following composition 2,2'-dithiodi-1H-imidazole 300 mg mass for suppositories 1,450 mg
The finely sprayed active substance is suspended in the mass for suppositories at 370C and the mixture is poured into molds which have been slightly cooled beforehand. Weight of one suppository: 1,750 mg.

Claims (3)

CLAIM 1. Pharmaceutical composition for the inhibition of malformation or destruction of erythrocytes due to a genetic modification of hemoglobin or to parasites, characterized in that it contains, as active principle, a compound of formula

 in which R represents hydrogen, R 'represents an NH2 group or else R and R', taken with the nitrogen and carbon atoms to which they are attached, form a radical 1H-imidazol-2-yl, 1-methyl1H-imidazol -2-yl, 4,5-dihydro-1H-imidazol-2-yl, 4,5-dihydro-1methyl-1H-imidazol-2-yl, thiazol-2-yl, 4,5-dihydrothiazol-2-yl , 1H-1,2,4-triazol-3-yl, 1-methyl-1H-1,2,4-triazol-3-yl or pyrimidin 2-yl, optionally in the form of a pharmaceutically acceptable salt. 2. Coinposition according to claim 1, characterized in that it is in the form of metering unit. 3. Composition according to claim 2, characterized in that each dosage unit contains from 10 to 1000 mg of active principle.