LU84808A1 - ENANTIOMERS OF HEXAHYDRO-2,3,3A, 4,5,6 1H-INDOLO (3,2-1-DE) (NAPHTYRIDINE-1,5), THEIR SEPARATION PROCESS AND THEIR APPLICATION IN THERAPEUTICS - Google Patents

Description

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The present invention relates to the enantiomers ce he-xahycrc-2,3,3a, 4,5,6 ΙΚ-inöclo [3, 2,1 -dej [n aphtyri cine-1, Si, their addition salts with pharmaceutically acceptable acids their oresration and their therapeutic application.

r * · * 5 Luxembourg patent No. 83,775 describes, among other things, the racemate that hexahydro-2,3,3a, 4,5,6 IH-incolo [3,2,1-de] [naphthricricin-1 , 5], which corresponds to formula (I)

(Z NH

mrVS

In the structure of this compound, the carbon atom in position 3a is asymmetrically substituted, which gives rise to two optical antipodes.

Each of these enantiomers can be isolated thanks to the intermediate preparation of the addition salt to one of the optically active antipodes of a chiral acid such as bis 15 (4-methyl-benzoyloxy) - 2,3 butanedioic acid, 1 * bromo-3 camphorsulf onic acid-.δ or-10, camphorsulf onic acid-l 0, tartaric acid, N - (<x-methylbenzyl) monoamice ce phthalic acid, maiic acid or l mancéliçue acid.

Salification takes place between room temperature and the boiling temperature of the solvent, using a mixture of enantiomers of formula (I), for example from racemate, in the form of this base, dissolved in a porous solvent such as an alcohol, preferably ethanol.

The two ciastereoisomeric salts thus obtained are then separated by fractional crystallization.

After separation, they can be separated from this acid / chiral, then returned to the form of an addition salt, starting from a base partially enriched in one of the enantiomers, originating for example from mother liquors of recrystallization from 'a mixture of salts or as téréciscmëres. 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 10 (CPV) or liquid chromatography (CLE?) According to a method described with regard to amino acids because 3. Halpern in Handbook of Derivatives for Chromatography, pp . 457-476, edited by K. Blau and G.S. King at Heyden.

The method consists in condensing the isomer this formula (I) 15 with the chloroformiere of {-) menthyl, of formula (II), prepared according to the method described in the cited work, d5! "A (II) C Cl

CH CW

C 3 3 so as to obtain the corresponding menthyl carbamate. The latter, examined by CPV or CLEP chromatography, gives rise to a single peak if there is indeed only one enantiomer of formula (I), and to two peaks if the resolution is incomplete.

The following examples illustrate the separation of these isomers from the compound this formula (I), and the verification of their purity.

Γ Ί EXAMPLE 1 (+) -hexahydrc-2,3,3a, 4,5,6 IH-indolo! 3,2,1-cs Γ 1 25 naphthyridine-1,5! and its methanesulfor.ate.

^ - i. ion je salts disse Précisorèjoas with arid '- / - ~ ^ -, § t h v 1-4 b e n r oy1c z y · - 2,3 buranedioïque.

_ or · 1 afTcVgr of 2 i §- ”iré d'tr. ari ratetr mecnéticue,

CL

,, _.-â'ri-éranc = reflux 5r c'ur. oil bath heating, G UH - · *, - ,, β 34.41 g of hexahy ~ r0 - 2,3,3a, 4,5,6 1 H - indolo D or * pi -'- '' - (--1 | 3 21 -de j naphthyridine -1.5 racemic under base horn, .η .- "obtained according to example 1 of the oily patent application

n ° 8024717 'sn solution in 250 ml of absolute ethanol. AT

"'. -A-e-ambient, and while stirring, we add slowly- tempera t,? 73 σ of acid (t) -bis (4-methvi-benzoyloxy) - 2,3 10 ment J- '' - - "p-io" cue in solution in 250 ml of absolute ethanol. It abuts “0-1 · - there is an abundant crystallization before the end -1, - -ai <-ion. The mixture is then heated slowly to 1 at reflux temperature, cooked, then another 15 200 ml of ethanol is added to obtain complete dissolution of: crystals · Elm ^ due to some insoluble crystals by filtration, the filtrate is heated and it is allowed to cool to room temperature. After a few hours of rest, the Iss crystals cormes are filtered on sintered glass, and they are dried under vacuum at 50 ° C. 43.56 g of white crystals are collected.

b) Recrystallization

Several crystals are recrystallized from the crystals thus obtained in 95% etnanoi. After each recrystallization, approximately 0.25 g of crystals are removed, the base of which is released by a mixture of dilute ammonia and this chloroform, and the rotary power is determined. At the end of cuatre • r write-because nsa hold the rotary power qp the base remains stable at around 30 LJd = -r 63.29 (c = i in MeOH).

The salt from which this base was released then has a rotary power of about // \ L ^ JD = - 84.51 °! C = l in MeCH).

5.73 g of the salt obtained according to b) are placed in a 250 ml erisrmeysr fitted with a macrnetic stirrer with 100 ml of 10¾ ammonia and 100 ml of ethyl acetate. After stirring for a few minutes, the organic phase is separated, the aqueous phase is taken up in 100 ml of ethyl acetate, the two organic phases obtained are combined, washed with water, dried over sulphate. magnesium, they are concentrated in a rotary evaporator 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 methanesulfonic acid, ie 1.34 g, is added, dissolved in 15 ml of ethanol absolute. An abundant crystallization is immediately formed. The mixture is further stirred for half an hour at room temperature, then the crystals are filtered on sintered glass and dried under vacuum at 50 ° C. Recrystallization from 60 ml of hot absolute ethanol, followed by an overnight rest at room temperature provides, after filtration and drying under defect, 3.03 g of methanesulfonate crystals which melt at 222-224 ° C. Rotatory power: J 1 20 = + 21.58 ° (c = l in MeOK)

JD

25 Elementary analysis and the RKN and IF spectra. confirm the structure of the compound.

d) Verification of purity.

140 mg of (^) - hexahy-30 dro-2,3,3a, 4,5,6 IH-indoloj3, are placed in a 100 ml flask fitted with a magnetic stirrer and a calcium chloride guard. 2,1-deJ j ^ naphtyridir.e-1,5 eu solution in 10 ml of ethyl acetate with 66.7 mg (1 equivalent.) This triethylamine, and the mixture is cooled in an ice bath and of water. We then add an equivalent of '-) - cnxororcr-ft miats de nentnvie in solution in toluene, that is to say 1 ml to v \

_ AT

6.6 Mo / mi CI, and gold. continue stirring 15 minutes at 0 ° C and then 20 minutes at room temperature. The insoluble material is removed by filtration, and the organic phase is washed with water, then with a 5% aqueous solution of this sodium bicarbonate, then again with this water. Then gold. dry on sulfate ce; magnesium, it is concentrated in a rotary evaporator then I under vacuum. We get 251 me of a yellowish oil which we | prepare a solution of 0.1 mg / ml in ethyl acetate.

l | Gas chromatography of this solution provides a curve showing a single peak.

: (| EXAMPLE 2 (-) -hexahydrc-2,3,3a, 4,5,6 IH-indolo | 3,2,1-de J

¢: Γ -η “LJ

►j naphthyridine-1.5 I? [

P

II 'a) release of the base from the mother liquor of the recrystallization of the dextrorotatory isomer in the form of \ 15 bis (4-methvl-2,3-benzovloxvj-2,3 butanedioate.

i; i‘j jj In a rotary evaporator, the filtrate of the? First recrystallization carried out according to example 1b).

S.

! Then add 200 ml of 10% ammonia, then 200 ml of ethyl acetate. We have the melance tending cuel- | I - "| 20 eues minutes then it is filtered on sintered glass.

They are organic phase, washed with water, dried over magnesium sulphate and concentrated in a rotary evaporator and then under vacuum. This gives approximately 9.11 g of an orange oil.

B) formation of diastereoisomeric salts with the acid (-} - bis I _ --- a imethvl-4 benzovloxv ^ -2,3 butar.ecioiaue.

--------- I 'Ü 0 In a 500 ml Erlenmever fitted with a magnetic stirrer 1 “" ü and a re-cooler and placed in an oil bath, we cook 9 0.08 g of the oil obtained according to Example 2a) dissolved pi 3 0 in 65 ml of absolute ethanol then, at room temperature and ï.} Ij over the course of an hour, a solution of 'acid f! (-) - bis (methvl - 4 ber.zovloxv) - 2,3 bufcanediorcue in: I / 65 ml of absolute ethanol. An abundant crystallization occurs i "before the end of the addition. Next we heat! slowly mix the mixture until this reflux is reached, and 140 ml of absolute ethanol are added to obtain the cissclu-; total crystals. One eliminates eueloues crystals insc-! ** *; 5 fats by filtration, the filtras are heated and left to stand at room temperature. After crystallization, Ί is filtered on sintered glass and dried under vacuum at 50 ° C. 11.2 g of this white crystals are thus collected.

'1; i

Il c) tecristall Isa tion il il il 10 The crystals thus obtained are recrystallized several times: l in 95% ethenol. Acres chaaue recrystallization on! * * I sn taken approximately 0.25 g of which the base is released by means of ammonia and chloroform, in order to determine the power: see rotary. After five recrystallizations, the rotational power of the base remains stable at around U / Ί = -63.38 ° 4 (c = 1.39 in MeOH). The salt then has a rotary power 1 of approximately [c ^ ^ = -89.21 ° (c = i in MeOH).

I EXAMPLE 3 (-) - hexahydro-2,3,3a, 4,5,6 IH-indolo j3,2, l-d ^ J

Ij J naphthyridine-1.51 and its methanesulfonate.

'' ί! 120 a) formation of diastereoisomeric salts with acid (-) - bis (methyl - 4 benzoyloxv) -2,3 butanediolgue

In a similar way to that in example la, we react | * 34.41 g of oily base in solution in 250 ml of ethanol d absolute, with 32.78 g of acid (-) - bis (4-methyl-benzoyloxy) '25 -2.3 butanedioic acid in solution in 250 ml of absolute ethanol, i * all in a 2 1 erlennsyer with magnetic agitation,! reflux refrigerant and oil bath heating. A criscal- | abundant addition occurs 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 in order to obtain dissolution. Some insoluble crystals are removed by filtration. We reheat the filter and we leave it -j - let reoeser at room temperature during the crystal-: S / ~ \ / _ --icts and read, after which we return to green- *. c> 3 5 f d ^ ^ ss vacuum drying at 50 ° C. Gn collects aïo-- crystals.

b) recrystallization 5 We proceed COITuTiS in Example 1 c). -'-? r. the base remains recrystallizations, the rotary power ^ * Γ 71 ”it salt at àlçxlp = _- 60.5 (c = l, l5 in MeOH) and Cw approximately [p (j D = - 87.9 ° (c = 1 in MeOH) - c) methanesulfonate 10 In a 250 ml flask fitted with a 3 ^ -01 ^ agil, 1/15 g of the enantiomer (oil) and solution are placed in 2q ml of ethyl ether and · 1 equivalent is quickly added, that is to say 0.52 g of methanesulfonxrous acid dissolved in 6 ml of ethanol. Crystals are formed, which is continued to stir for half an hour before wringing them out and drying them under vacuum at 50 ° C. Then they are recrystallized from 19 ml of absolute ethanol. 1.12 g of methanesulfonate are collected, which melts at 219-221 ° C.

25

Rotatory power: j ^ j = ~ 22.0 ° (c. = 1 in MeOH).

Elemental analysis and NMR and IR spectra confirm the structure of the compound.

d) purity check

The procedure is as in Example 1 d. The carbamate this r.enthyle obtained gives, in gas chromatography, a 21 curve orssing a single goose.

U

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The compounds of the invention have been subjected to pharmacological tests intended to demonstrate their interest in anerapeutians.

Hypoxia hvnobare 5 CD strain mice! are maintained in an oxygen-depleted atmosphere, by producing a partial vacuum (190 mm of mercury corresponding to 5.25% of oxygen).

The survival time of the animals is noted. This time is increased by agents capable of promoting oxygenation of the tissue and in particular of the brain. The test compounds are administered, in several doses, intraperitoneally, 10 minutes before the test. The percentages of increase in survival time compared to the values obtained in the control animals are calculated. The average active dose which increases the survival time by 100% (DA10q) is determined graphically. The is from 21 to 23 mg / kg. The AD ^ q is 7 to 10 mg / kg.

Global Ischemia Test in Mice The survival time of the test animals is measured after having injected 0.1 ml of a saturated magnesium chloride solution into the tail vein. The resulting cardiac arrest causes cerebral ischemia. "Survival time" * is the time interval between the injection of magnesium chloride and the last inspiratory movement of each mouse, considered to be 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 magnesium chloride, are compared with the survival times of these control animals to which no ad- / minister that the vehicle these active substances.

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* '<, Mice studied by rump of 10, the averages of the results of each group make it possible to draw a curve, thanks to which the effective dose is graphically determined, expressed in mg of active substance per kg of body weight, which extend this survival time for 3 seconds (DE-)

An increase in survival time of 3 seconds is both statistically significant and reproducible.

The DE2 of the compounds of the invention are from 7 to 10 mg / kg.

The pharmacological study of the compounds of the invention shows that they have an antianoxic activity and that they can be used in therapy for the treatment of disorders of alertness, in particular for combating the behavioral disorders attributable to this damage. cerebrovascular and cerebral sclerosis, in geriatrics, as well as for the treatment of absences due to head trauma, for the treatment of these metabolic encephalopathies and for the treatment of depressive states.

The invention therefore includes all pharmaceutical compositions containing the compounds of the invention or their salts as active ingredients, in combination with any excipients suitable for their administration, in particular by oral or parenteral route.

* The routes of administration can be the oral and parenteral routes.

* »25 The daily dosage can range from 10 to 100 mg.

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Claims (5)

1. Process for the separation of the enantiomers of hexahvcrc- Ί Γ,. 71; 2,3,3a, 4,5,6 IH-indolo | 3,2, i-dej jjiapr.tyricrne-1, m, charac terized in this eus d! Cn raid rsacir un —ancance of 1 enantiomers of formula (I ) ii '"-Z r in base form, with one of these enantiomers of a chiral acid, one separates one of these two diastereoisomeric salts thus obtained by fractional crystallization and it is returned to base form and, if if desired, the latter is converted into an acceptable addition salt to a pharmaceutically acceptable acid * * 2. The process according to claim 1, characterized in that the chiral acid is bis (4-methylbenzovloxy) acid - 2,3! | Butanedioic J 15 3. Method according to one of claims 1 and 2rcaractérisé i in that the mixture of enantiomers of rorniule (I) is the racemate I. 4. Method according to one of claims 1 and 2, characterized in that the mixture of enantiomers of formula (I) consists of the mother liquors of a previous separation. 4 5. Method according to any one of claims 1 to 4, characterized in that salification is carried out with chiral acid and recrystallization from ethanol. i _6. La (-J-) -hexahydro-2,3,3a, 4,5,6 1K indolo, 2,1-dej I naphthyridine-1,5; and its addition salts with acids 1 L— ->! nharmaceuticuemenc acceptable. l / \ *; · - '................. Γ .1 / · ν; ..; r, -i, j, ^,,, c ..: *. r; GC_o j ^ i, Z. f 1 ce j: - naphtyriàir.s-1,5; and its aid salts with pharra- acids! There! ceuticue.T.enr aceeotables. 8. Coir.T3osit.iori pharraceuticus, characterized er. that it: 5 contains a corrocated according to any one of the claims; 6 and Ί. t i: <| ij 5 Ji | ! j i - i = * i i