FR2527210A1 - ENANTIOMERS OF HEXAHYDRO-2,3,3A, 4,5,6 1H-INDOLO (3,2,1-DE) (NAPHTHYRIDINE-1,5), THEIR SEPARATION METHOD AND THEIR THERAPEUTIC APPLICATION - Google Patents

Abstract

ENANTIOMERS RESPONDING TO THE FORMULA: (CF DRAWING IN BOPI) ARE ISOLATED FROM THEIR MIXTURE BY PREPARING THEIR ADDITIONAL SALTS TO AN ENANTIOMER OF A CHIRAL ACID. THE DIASTEREOISOMERS OBTAINED ARE SEPARATED BY FRACTIONAL CRYSTALLIZATION, THEN RETURNED IN THE FORM OF BASE OR PHARMACEUTICALLY ACCEPTABLE SALTS. APPLICATION IN THERAPEUTICS.

Description

25272 10

  The present invention relates to the enantiomers of the

  xahydro-2,3,3a, 4,5,6 l H-indolo L 3,2,1-deilnaphthyridine-1,5 l

  their pharmaceutically acceptable acid addition salts

  their preparation and their application in therapeutics.

  French Patent Application No. 80 24717 describes, among others, the racemate of 2,3,3a hexahydro, 4,5,61 H-indolo, 2,1-del naphthyridine-1,5, which has the formula (1) 3 NH 3 1I) In the structure of this compound, the carbon atom at position 3a is substituted asymmetrically, which gives rise to

with two optical antipodes.

  Each of the two enantiomers can be isolated by means of

  intermediate partioning of the addition salt to one of the optically active antipodes of a chiral acid such as bis (methyl 4 benzoyloxy) 2,3 butanediol acid, 3-bromo-camphorsulfonic acid-8 or -10, camphorsulfonic acid-10,

  tartaric acid, N- (c-methylbenzyl) monoamide of

  phthalic acid, malic acid or mandelic acid.

  Salification is between room temperature and the boiling point of the solvent, from a mixture

  enantiomers of formula (I), for example from the raceme

  mate, in basic form, dissolved in a suitable solvent

  such as alcohol, preferably ethanol.

  The two diastereoisomeric salts thus obtained are then

  separated by fractional crystallization.

  After separation, they can be separated from the chiral acid and then returned as an addition salt to any

physiologically acceptable acid.

  An enantiomer of formula (I) can also be obtained at

  from a partially enriched base to one of the en

  from, for example, recrystalline mother liquors

  The use of a mixture of diastereoisomeric salts The chiral acid used for the separation of such an enriched mixture is then the antipode of that which made it possible to obtain said mixture. The purity of the optical isomers of formula (I) can be verified for example by gas chromatography (CPV) or liquid chromatography (HPLC) according to a method described for amino acids by B Halpern in Handbook of Derivatives for Chromatography, pp. 457-476. , edited by K Bla U and

G.S. King at Heyden.

  The method consists in condensing the isomer of formula (I) with (-) menthyl chloroformate, of formula (II), prepared according to the method described in the cited work, O Cl (II)

CH 3 CH 3

  so as to obtain the corresponding menthyl carbamate.

  The latter, examined by CPV or HPLC chromatography, gives rise to a single peak if one is in the presence of a single

  enantiomer of formula (I), and at two peaks if the

is incomplete.

  The following examples illustrate the separation of isomers

  of the compound of formula (I), and the verification of their purity.

  EXAMPLE 1 (+) - 2,3,3a hexahydro, 4,5,6H-indolo 3,2,1-d

  lnaphthyridine-I, and its methanesulfonate.

  a) Formation of diastereoisomeric salts with (+) -

  bis (4-methylbenzoylozy) 2,3 butanedioic acid.

  In a 2 liter Erlenmeyer flask equipped with magnetic stirrer, reflux condenser and oil bath heating, 34.41 g of hexahydro 2,3,3 a, 4,5,6 1 H indolo l 3.21 -dellnaphthyridine -1.5 l racemic in oily base form obtained according to Example 1 of Patent Application No. 8024717, dissolved in 250 ml of absolute ethanol A

  room temperature, and while stirring, slowly add

  32.78 g of (+) - bis (4-methyl-benzoyloxy) -2,3-butanediolic acid in solution in 250 ml of absolute ethanol. An abundant crystallization occurs before the end of the addition. the mixture to the reflux temperature, then another 200 ml of ethanol is added to obtain the complete dissolution of the

  crystals Some insoluble crystals are removed by

  The filtrate is warmed up and allowed to cool to room temperature. After a few hours of rest,

  filters crystals formed on sintered glass, and separates them

  Vacuum at 50 ° C. 43.56 g of white crystals are collected. b) Recrystallization The crystals thus obtained are recrystallized several times in 95% ethanol. After each recrystallization, approximately 0.25 g of crystals are taken, the base of which is liberated with a mixture of dilute ammonia and chloroform, and the rotatory power is determined After four recrystallizations, the rotatory power of the base remains stable at approximately

2

LD = + 63.290 (c = 1 in MeOH).

  The salt from which this base was released then has a rotatory power of about

LOD = + 84.51 (c = 1 in MeOH).

c) Methanesulfonate.

  In a 250 ml Erlenmeyer flask equipped with a magnetic stirrer

  5.78 g of the salt obtained according to b) are placed with 100 ml of ammonia.

  10% and 100 ml of ethyl acetate After stirring for a few minutes, the organic phase is separated off, the aqueous phase is taken up in 100 ml of ethyl acetate, the two organic phases obtained are combined, wash with water, they are

  dried over magnesium sulphate, they are concentrated in a

  rotary porator and dried under vacuum The base is thus collected in the form of 2.96 g of a mixture of white crystals

and an oil.

  In a 250 ml flask, with magnetic stirring, this base is dissolved in 60 ml of ethyl ether and the equivalent quantity of methanesulphonic acid, ie 1.34 g, dissolved in 15 ml of absolute ethanol is added thereto. an abundant crystallization is immediately formed. The mixture is stirred for a further half hour at room temperature and then the crystals are filtered off.

  on sintered glass and dried under vacuum at 50 ° C. A recrystallizer

  in 60 ml of hot absolute ethanol, followed by a night of rest at room temperature, after filtration and drying in vacuo, 3.03 g of methanesulphonate crystals which mp 222-224 ° C. Rotatory power: ll = + 21.58 (c = 1 in MeOH) The elemental analysis and the NMR and IR spectra confirm the

structure of the compound.

d) Verification of purity.

  In a 100 ml flask equipped with a magnetic stirrer and

  140 mg of (+) - hexahydrate are placed in a calcium chloride

  2,3,3-a, 4,5,6 l H-indolol 3,2,1-del lnaphthyridine-1,5 len

  in 10 ml of ethyl acetate with 66.7 mg (1 equivalent) of triethylamine, and the mixture is cooled in a bath of

  ice and water Then add an equivalent of (-) - chloroform

  menthylmilate dissolved in toluene, ie 1 ml at 6.6 × 10 -4 mol / ml, and stirring is continued for 15 minutes at room temperature.

  0 C then 20 minutes at room temperature.

  soluble by filtration, and the organic phase is washed with water and then with a 5% aqueous sodium bicarbonate solution, then again with water Then it is dried over magnesium sulfate, concentrated in an evaporator then rotated under vacuum 251 mg of a yellowish oil is obtained.

  prepare a solution of 0.1 mg / ml in ethyl acetate.

  The gas chromatography of this solution provides

a curve with a single peak.

  EXAMPLE 2 (-) -hexahydro-2,3,3a, 4,5,6-LH-indolo [3,2,1-del] naphthyridine-1,5a) liberation of the base from the recrystallization mother liquor of the dextrorotatory isomer in the form of

  bis (4-methylbenzoyloxy) -2,3 butanedioate.

  In a rotary evaporator the filtrate of the

  first recrystallization carried out according to Example 1b).

  It is then added 200 ml of 10% ammonia, then

  ml of ethyl acetate The mixture is stirred for a period of

  The organic phase is separated off, washed with water, dried over magnesium sulphate and concentrated in a rotary evaporator and then under vacuum. The result is thus obtained approximately 9.11 g.

an orange oil.

  b) formation of diastereoisomeric salts with (-) bis acid

  (4-methyl-benzoyloxy) -2,3-butanedioic acid.

  In a 500 ml Erlenmeyer flask equipped with a magnetic stirrer

  and a refrigerant and placed in an oil bath is introduced

  9.08 g of the oil obtained according to Example 2 a) are dissolved in 65 ml of absolute ethanol and then, at room temperature and over one hour, an acid solution is added thereto (- ) bis (methyl 4 benzoyloxy) 2,3 butanediol in ml absolute ethanol Abundant crystallization occurs even before the end of the addition Then the mixture is slowly heated to reflux temperature, and

  140 ml of absolute ethanol are added to give the dissolu-

  The crystals are removed completely.

  By filtration, the filtrate is warmed up and allowed to stand at room temperature. After crystallization, it is filtered on sintered glass and dried under vacuum at 50 ° C.

  thus collects 11.2 g of white crystals.

  c) recrystallization The crystals thus obtained are recrystallized several times in 95% ethenol. After each recrystallization, approximately 0.25 g is taken off and the base is liberated by means of

  ammonia and chloroform, in order to determine the

  At the end of five recrystallizations, the

  the rotation of the base remains stable at about lol = -63.38 (c = 1.39 in MeOH). The salt then has a rotatory power.

  about 1 = -89.21 (c = 1 in MeOH).

  EXAMPLE 3 (-) hexahydro-2,3,3a, 4,5,61 H-indolo l 3,2,1-d

  lnaphthyridine-1,1 and its methanesulfonate.

  a) formation of diastereoisomeric salts with (-) bis (methyl 4-benzoyloxy) -2,3-butanedioic acid Analogous to that of example 1a, 34.41 g of oily base are reacted in solution in 250 ml of absolute ethanol, with 32.78 g of (-) bis (4-methyl-benzoyloxy) -2,3-butanediolic acid in solution in 250 ml of absolute ethanol, all in a 2-liter Erlenmeyer flask with magnetic stirring,

  reflux condenser and oil-bath heating A crystal-

  The abundant solution is produced before the end of the addition of the diacid at room temperature. The mixture is then heated to reflux and 220 ml of absolute ethanol are added to the mixture to obtain the dissolution. Some insoluble crystals are removed by filtration. the filtrate and we

  let stand at room temperature for the crystal

  After this, the mixture is filtered on sintered glass and dried in vacuo at 50 ° C. and 35.47 g of crystals are collected. b) recrystallization The procedure is as in Example 1c) After four recrystallizations, the rotatory power of the base remains at pH D = 60.5 (c = 1.15 in MeOH) and that of the salt to

  about D = 87.9 (c = 1 in MeOH).

  c) methanesulphonate In a 250 ml flask fitted with magnetic stirring, 1.15 g of the enantiomer (oil) in solution in

  ml of ethyl ether and the equiva-

  0.52 g of methanesulphonic acid in solution in m of ethanol is formed. Crystals are formed, which are stirred for half an hour before being wrung and dried under vacuum at 50.degree. C. They are then recrystallized in 19 ml of absolute ethanol. Finally, 1, 12 g is collected.

  of methanesulfonate which melt at 219-221 C.

  Rotatory power: lEl = 22.0 (= 1 in MeOH).

  D Elemental analysis and NMR and IR spectra confirm

the structure of the compound.

  d) Verification of the Purity One proceeds as in Example 1 d The menthyl carbamate obtained gives, in gas chromatography, a

21 curve with a single peak.

  The compounds of the invention have been subjected to phar-

  to highlight their interest in therapeutics. Hypobaric Hypoxia CD1 strain mice are maintained in a depleted oxygen atmosphere, by performing a partial vacuum

  (190 mm of mercury corresponding to 5.25% of oxygen).

  The survival time of animals is noted This time is increased

  agents capable of promoting tissue oxygenation.

  The compounds studied are administered, at several doses, intraperitoneally, minutes before the test. The percentages of increase of the survival time with respect to the values obtained in the control animals are calculated. The mean active dose which increases the survival time of 100% (DA 100) is determined graphically. The DA 100 is 21 to 23 mg / kg. The DA 50 is

from 7 to 10 mg / kg.

  Overall Ischemia Test in the Mouse The survival time of the test animals was measured after injecting 0.1 ml of a saturated solution of magnesium chloride into the tail vein. The resulting cardiac arrest causes "cerebral ischemia" The "survival time" is the time interval between the injection of magnesium chloride and the last inspiratory movement of each mouse, considered the ultimate index of a function of the

central nervous system.

  The survival times of the animals treated with the

  compounds of the invention administered intraperitoneally

  10 minutes before the injection of the magnesium chloride, and

  the survival times of the control animals to which no

  only the vehicle of the active substances.

  The mice being studied in groups of 10, the averages of the results of each group make it possible to draw a curve, by which the Effective Dose, expressed in mg of active substance per kg of body weight, which prolongs the survival time, is graphically determined. of 3 seconds (DE 3) An increase in the survival time of 3 seconds is at the

  once significant statistically and reproducibly.

  DE 3 of the compounds of the invention are 7 to 10 mg / kg.

  The pharmacological study of the compounds of the invention shows that they possess an anti-anoxic activity and that they can be used therapeutically for the treatment of the disorders

  vigilance, in particular to fight against

  behavioral problems attributable to cerebrovascular disease and cerebral sclerosis, in geriatrics, as well as

  for the treatment of absences due to traumatic cru-

  for the treatment of metabolic encephalopathies

  and for the treatment of depressive states.

  The invention therefore includes any pharmaceutical compositions

  containing the compounds of the invention or their

  salts as active ingredients, in association with all exci-

  appropriate for their administration, in particular by

oral or parenteral.

  The routes of administration may be the oral and parenteral routes.

  The daily dosage can range from 10 to 100 mg.

Claims (8)

claims   1 Process for separating enantiomers from hexahydro-   2,3,3 a, 4,5,6 l H-indolo L, 2,1-d Lnaohtyridine-t, 5,   in that a mixture of enantiomers of formula (I) in the form of a base is reacted with one of the enantiomers of a chiral acid, one of the two diastereoisomeric salts thus obtained is separated by fractional crystallization and it is returned to the base form and, if desired, converted into a pharmaceutically acceptable acid addition salt acceptable.   2 Process according to claim 1, characterized in that the chiral acid is bis (4-methyl-benzoyloxy) -2,3-butanediol acid.   3 Method according to one of claims 1 and 2, characterized   in that the mixture of enantiomers of formula (I) is the racemate.   4 Process according to one of claims 1 and 2, characterized   in that the mixture of enantiomers of formula (I) is   constituted by the mother waters of a previous separation.   Process according to any one of claims 1 to 4,   characterized in that salification is carried out with   chiral acid and recrystallization from ethanol.   (+) 2,3,3-hexahydro-4,5,6 1 H indolo l 3,2,1-naphthyridine-1,1 and its addition salts with acids pharmaceutically acceptable.   7 (-) - hexahydro-2,3,3 a, 4,5,6 1 H-indolo l 3,2,1-de   lnaphthyridine-1, and its addition salts with pharmaceutical acids ceutically acceptable.   8 Pharmaceutical composition, characterized in that   contains a compound according to any one of the claims 6 and 7