FR2722199A1 - New valproic acid ester(s) esp. of saccharide cpds. - Google Patents

Abstract

Valproic acid (VPA) ester derivs. of formula (I) W-(O-CO-CH(CH2CH2CH3)2)n are claimed. In (I), W is residue of cpd. (II) having at least one OH gp. esterifiable by VPA or labile part Y which can be substd. by a valproate gp.; and n is 1 to total number of esterifiable OH gps. or substitutable Y gps. in (II); with the proviso that W is derived from e.g 1,3-di-(palmitoyl, decanoyl, hexanoyl, butanoyl or acetyl)-2-valproylglycerol; 2-valproylglycerol, 1,3-divalproylglycerol; and 2-(acetyl, butanoyl or decanoyl)-1,3-divalpropylglycerol etc.. (II) is e.g. monosaccharide, polysaccharide or itol.

Description

Derivatives of valproic acid and their applications as drugs

  The present invention relates to valproic acid derivatives and their applications.

as medicines.

  It is known that valproic acid (2-propylpentanoic acid or dipropylacetic acid) and its salts and valpromide (2propylpentanamide or dipropylacetamide) possess antiepileptic properties. Valproic acid and its salts have a broad antiepileptic spectrum, they are used in the treatment of generalized epilepsies of the type Grand Mal, Petit Mal and myoclonic. They are also used in pediatrics for the treatment of convulsions

  fever and to prevent seizures in high-risk children.

  Valpromide versus valproic acid has superior anticonvulsant properties, but its overall toxicity and neurotoxicity are higher. For these reasons it is prescribed in the case of rebellious epilepsies but it is not recommended in pediatrics. The treatment of epilepsies is obligatorily chronic and the two molecules currently used in therapeutics, Depakine (valproic acid and its sodium salt) and Depamide (valpromide), require the taking of several daily doses; this causes, given their short plasma half-life, a significant fluctuation in their blood concentration between two doses. The consequences of this fluctuation are, on the one hand, less protection after a few hours and, on the other hand, a risk of overdose, just after taking, favoring side effects. The Applicant has set itself the goal of synthesizing chemical compounds containing in their structure at least one valroyl radical bonded, by a covalent bond, to an atoxic carrier compound so that these compounds, in-vivo, have a biological activity. after release or not of valproic acid with result as a longer half-life than that of saline valproates or amide derivatives above and allowing an extension of the antiepileptic effect and a lower variation of serum levels reducing so

the side effects.

  This object is achieved by the compounds according to the invention of formula W - (O-COCH (-CH 2 -CH 2 -CH 3) 2) n in which W represents a radical of a product having at least one ester-esterified OH group by the valproic acid or its derivatives or alternatively, a leaving part Y which may be substituted by the valproate group, n being able to vary from I to the total number of esterifiable OH groups or Y

substitutable in said product.

  In the context of the present invention, the radical product W acts as a valproic acid transporter and it is particularly advantageous to choose it from the least toxic products possible with respect to living organisms, in the extent o

  product is released into the body and must therefore be eliminated or metabolized.

  In a preferred manner, the ester or polyester compounds of the present invention will have a radical W having the precursor W-OH which may be a monosaccharide, a polysaccharide

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  or else an itol, functionalized or not, protected or not, and preferably chosen from

  D-glucose or its derivatives 1,2: 5,6-diacetoneglucose or 1,2: 3,5diacetoncglucose or 1,2-

  monoacetoneglucose, D-galactose or diacetonegalactose, Dfructose or its derivatives

  2,3,4,5-diacetonefructose or 1,2,4,5-diacetonefructose or 1,2monoacetonefructose or 2,3-diacetonefructose

  monoacetonefructose, D-mannose or its derivatives 2,3: 5,6-diacetonemannose or 2,3-

  monoacetonemannose, D-allose or its derivatives 1,2: 5,6-diacetonealllose or 1,2-

  monoacetone, D-xylose or 1,2-monoacetonexylose, xylitol or its derivatives 2,3: 4,5-

  diacetonexylitol or 2,3-monoacetonexylitol or 4,5-monoacetonexylitol or 2,4: 3,5-

  dimethylènexylitol or from disaccharides and polysaccharides that may result from the combination of two or more of the monosaccharides previously mentioned. Are excluded as

  precursor glycerol and solketal.

  The valproyl group or groups are grafted onto W either by esterification of one or more free OHs of the W-OH molecule or by substitution of one or more groups

  Carried by W according to the general methods of literature.

  The compounds according to the present invention are intended for use as a medicament alone or in combination with other compounds, such as valproic acid and its salts or amides, for example, for relaxing and / or hypnotic use and / or

  anticonvulsant and / or antiepileptic drug.

  In addition to the foregoing, the invention also comprises other provisions,

  which will emerge from the following description. It must be understood, however, that this

  description and the examples it contains are for illustration purposes only.

  the object of which they do not constitute in any way a limitation.

  Example n l. Preparation of 2-Propylpentanoic Acid Chloride

valproyle).

  In a three-necked flask of 500 ml, at room temperature, 30 g (2.5 × 10 -3) are introduced.

  mol) of thionyl chloride and then, dropwise, 30 g (2.1.10-1 mol) of 2-

  Propylpentanoic. At the end of the addition, the mixture is refluxed. After 30 minutes, the control CPV (OV17 column at 120 C length 75 cm) shows the total disappearance of valpr-oic acid. The distillation of the crude product makes it possible to eliminate successively the excess SOC12, as well as

  HCI and SOi formed and collect valproyl chloride (33.8 g yield: 98%).

  Example No. 2. Preparation of 3-O- (2'-propylpentanoyl) -1,2: 5,6-diisopropylidene

  a-D-glucofuranose (valproare of diaceroneglucose).

  In a knit flask was placed 32 g (1.2.10-1 mol) of 1,2: 5,6-di-Oisopropylidene-act-D-

  glucofuranose prepared by the known methods; 200 ml of chloroform, 15 ml of triethylamine (TEA) are then introduced and then dropwise, 22 g (1.4 × 10 -3 mol) of valproyl chloride dissolved beforehand in 100 ml of chloroform. We heat the whole

  with stirring until refluxing for 12h.

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  After returning the reaction mixture to ambient temperature, 250 ml are added.

  saturated aqueous solution of NaHCO3. The organic phase is decanted, washed successively with 2 × 150 mL of water, dried over anhydrous Na 2 SO 4 and then evaporated under reduced pressure. 54 g of reaction crude are obtained, which is purified by medium pressure chromatography on silica gel (Kielselgel 60, 500 ml). Is successively isolated: - after passage of 800 ml of hexane, 0.9 g (7.10-3 mol) of valproic acid; after passage of 1.5 L of hexane-ethyl ether mixture, 93: 7, (v / v), 30 g (yield: 63%) of diacetone glucose valproate which is in the form of a liquid

viscous pale yellow.

  [lt] O2 = -29.4 (c = 2.1; CHCl3) Crude formula: C20H3407 Elemental analysis: Calculated: C: 62.20%; H 8.90% Found: C: 62.40%; H: 8.90% The 1H and 13C NMR spectra are given in Tables A and B.

  Example No. 3 Preparation of 6-O- (2'-propylpentanoyl) -1,2; 3,4-di-O-isopropylidene

  α-D-galacropyranose (diacetonegalacrosis valproate).

  This compound is prepared from 1,2: 3,4-di-O-isopropylidene-α-D-galactopyranose according to the method of Example 2 with a yield of 74.8% after purification with

  medium pressure chromatography on silica gel [eluent: hexane-ethyl ether 93: 7 (v / v)].

  [X22 [a1] = -26.30 (c = 1.9; CHCl3) Crude formula: C20H3407 Elemental analysis: Calculated: C: 62.20%; H: 8.90% Found: C: 62.10%; H: 8.90% The RMIN H and 13C spectra are given in Tables C and D.

  Example No. 4 Preparation of 1-O- (2'-propylpentanoyl) -2,3,4,5-di-O-isopropylidene

  13-D-frucropyranose (diacetonefrucrose valproate).

  This compound is prepared from 2,3: 4,5-di-O-isopropylidene - [- D-fructopyranose according to the method of Example 2 with a yield of 56.1% after purification by

  medium pressure chromatography on silica gel [eluent: hexane-ethyl ether 93: 7 (v / v)].

  [C, 22 [a] 2 = -21.1 (c = 2.1; CHCl3) Crude formula: C20H3407 Elemental analysis: Calculated: C: 62.20%; H: 8.90% Found: C: 62.40%; H: 9.20% The spectra R.NIN IH and 13C are given in Tables E and F.

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  Example No. 05 Preparation of 1-0- (2'-propylpentanoyl) -2,3,4,5-diisopropylidene

  xylitol (valproaze of diaceronexylitol).

  This compound is prepared from 2,3: 4,5-di-O-isopropylidènexylitol according to the method of example 2 with a yield of 66.7% after purification by medium pressure chromatography on silica gel [eluent: hexane ethyl ether 93: 7 (v / v)]. [= -0.6 (c = 2.08, CHCl3) Crude formula: C19H340 Elemental analysis: Calculated: C: 63.70%; H: 9.60% Found: C: 63.60%; H: 9.70% The 1H and 13C NMR spectra are given in Tables G and H. Example No. 6. Preparation of 1-0- (2'-propylpentanoyl) -2,3: 5,6-di-O isopropylidene

  α-D-mannofuranoside (α-diacetonemannoside valproate) and 1-O- (2'propylpentanoyl)

  2,3: 5,6-di-O-isopropylidene-β-D-mannofuranoside (3-diacetonemannoside vaiproate).

  These compounds are prepared according to the method of Example 2, from a mixture of

  2,3: 5,6-di-O-isopropylidene-α-D-mannofuranose and 2,3: 5,6-di-O-isopropylidene-5-D-

  mannofuranose, in proportions ca / f = 75/25, also prepared according to the known methods. After purification of the crude by medium pressure chromatography on silica gel (Kielselgel 60), the following is isolated successively:

  after elution with hexane-ethyl ether mixture 93: 7 (v / v), the valproate of

  diacetonemannose, with a yield of 69.1% compared to the initial cadiacetonemannose.

  [OEID = -26.3 (c 1.59, CHCl3) Crude formula: C20H3407 Elemental analysis: Calculated: C: 62.20%; H: 8.90% Found: C: 63.20%; H: 9.30%

  after elurion with hexane-ethyl ether mixture, 88:12 (v / v), valproate I-

  diacetonemannoside with a yield of 80.6% compared to the initial [3diacetonemannose.

  Elemental analysis: Calculated: C, 62.20%; H: 8.90% - Found: C, 62.10%; H 9.10% The 1H and 13C NMR spectra are given in Tables J, K, L and M. Example No. 7. Preparation of 1-0- (2'-propylpentanoyl) -2,4: 3,5-di -O-methylene

  xylitol (dimethylennexylitol valproate).

  In a 500 ml three-neck flask containing 32 g (1.8 × 10 -3 mol) of dimethylenexylitol dissolved in 200 ml of pyridine, 38 g (2.4 × 10 -10 mol) of chloride are introduced dropwise at room temperature. of Valproyl in 100 mL of pyridine, and the mixture is refluxed. After 4:30 of stirring, the mixture is allowed to come to room temperature and then

  add 500 mL of water and 500 mL of hexane-ethyl ether, 50:50 (v / v).

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  The organic phase is decanted, washed successively with 200 ml of an aqueous solution of HCl (0.5 N) and twice with 200 ml of water, dried over anhydrous Na 2 SO 4, filtered and filtered.

evaporated under reduced pressure.

  The residue (72 g) is taken up in 500 ml of hexane and then brought to reflux with stirring. The residual dimethylennexylitol is filtered while hot and the filtrate is at 0 ° C. for 12 hours. After filtration, 40 g of white crystals are obtained which are recrystallized from 300 ml of hexane. 37 g (yield: 62.1%) of dimethylennexylitol valproate, which is in the form of white crystals, is isolated and has a melting point of mp = 86.5 ° C.

[c] 22 = -0.4 (c = 2.1, CHCl3).

  Elemental analysis: Calculated: C: 59.60%; H: 8.70% Found: C: 59.60%; H: 8.80% The spectra R..MN 1H and 13C are given in Tables N and P. The monoacetonide or non-ketalised compounds containing in their structure at least one valproyl radical can be prepared by partial or total deprotection of the compounds corresponding diacetonides, - either in homogeneous phase (organic solvent - H30 + or alkanol in an acid medium),

  - in the heterogeneous phase (acidic resin - organic solvent - H20 or acidic resin -

alkanol).

  Example No. 8 Preparation of 3-O- (2'-propylpenranoyl) -1,2-isopropylidene-a-D-

  glucofuranose (3-valproate monoaceroneglucose) and 3-O- (2'propylpentanoyl) -D-

  glucofuranose (glucose valproate).

  The compounds are prepared by passing 26 g of 3-O- (2'-propylpentanoyl) -1,2,5,6-

  di-O-isopropylidene-xD-glucofuranose in 10% (w / v) solution in dioxane-H2O solvent: 5 (v / v) on an Amberlist 15 H * wet resin column (250 mL), maintained at 70 C with a flow rate of 10 ml / min. After passing an additional solvent volume of 750 mL, the eluent is evaporated under reduced pressure. 25 g of crude reaction product are obtained, which is purified by

  medium pressure chromatography, silica gel.

  - Succession is obtained successively: after passing 400 ml of the hexane-ethyl ether mixture, 80:20 (v / v), 0.95 g of residual diacetoneglucose valproate; after passage of 400 ml of ethyl ether, 10 g of 3-valproate of

  monoacetoneglucose, which is in the form of a pale yellow viscous liquid.

  [aC] D2 = 9.00 (c = 1.5; CHCl3) Crude formula: C17H3007 Elemental analysis: Calculated C: 58.90%; H: 8.70% Found: C: 59.00%; H: 8, 90%

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  after passage of 400 ml of 50:50 (v / v) THF-ethyl ether mixture, 11 g of 3-

  glucose valproate in the form of white crystals, melt

F = 74.6 C.

  [] D2 = +55.1 (c = 1.90, CHCl3) Crude formula: C 14H2607 Elemental analysis: Calculated: C 54.90%; H: 8.60% Found: C: 53, 10%; H: 8.60% The proportion of 3-valproate of glucose can be increased when the above acid hydrolysis is carried out using a lower eluent flow rate in order to increase the contact time. Similarly, the monoacetonide compound can be obtained more selectively, when operating the acid hydrolysis, at a lower temperature or with a higher eluent flow rate,

  to reduce the contact time.

  The spectra R-M.N 1H and 13C are given in the tables Q-T.

  Example No. 9 Preparation of 6-0- (2'-propylpentanoyl) -α-D-galactopyranose

(galactose valproate).

  In a reactor thermostated at 45 ° C, place 80 ml of dioxane, 20 ml of solution

  aqueous 37% concentrated HCl and 10 g (2.6 × 10 -2 mol) of diacetone galactose valproate.

  After stirring for one hour, the mixture is cooled and the acid is neutralized by addition of 20 g of powdered Na.HCO3 and 20 ml of water. The organic phase is decanted and the aqueous phase reextracted with 50 ml of dioxane. The organic phases are combined, dried over anhydrous Na 2 SO 4, filtered and then evaporated under reduced pressure. The residue is taken up in 100 mL of THF

  then filtered and the filtrate evaporated under reduced pressure.

  10 g of crude reaction product are obtained which is purified by means of medium liquid chromatography.

pressure on silica gel.

  Is successively isolated: after passage of 300 ml of hexane-ethyl ether mixture, 80:20 (v / v), I g

  (2.6 × 10 -3 moles) of residual diaceronegalactose valproate.

  after passage of 300 ml of ethyl ether mixture -THF, 60:40 (v / v), 6.2 g

  (yield: 78.3%) of galactose valproate.

  [(c] 22 = +68.20 (c = 1.69, CHCl3) Crude formula: C14H2607 Elemental analysis: Calculated: C: 54.90%, H: 8.60% Found: C: 52.20%; H: 8.60% The RatN 13C spectrum is given in Table U.

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  Example No. 10 Preparation of 1-0- (2'-Propylpentanoyl) -2,3-Oisopropylid} - [- D-

  frucropyranose (monoacetone valproatefrucrose).

  In a reactor thermostatically controlled at 68 ° C., 25 μL of Amberlist 15H + wet acid resin and 4.5 g (1.16 × 10 -2 mol) of diacetonefructose valproate dissolved in 45 ml of ethanol-water mixture, 95: 5 ( v / v). After stirring for 6 hours, the mixture is cooled and then filtered. The filtrate

  is evaporated under reduced pressure and the residue obtained taken up in 100 mL of ethyl ether.

  The solution is dried over anhydrous Na 2 SO 4, filtered and then evaporated under reduced pressure.

  4.95 g of crude are obtained which is purified by medium pressure chromatography on silica gel. Is successively isolated: - after passing 500 ml of the hexane-ethyl ether mixture, 90:10 (v / v), 18 g of

  Residual diacetonefructose valproate.

  after passage of 500 ml of ethyl ether, 3.12 g (yield 69.2%) of valproate of

  monoacetonefructose which is in the form of a pale yellow viscous liquid.

  [o [] 22 = 7.410 (c = 2.8, CHCl3) Crude formula: C17H3007 Elemental analysis: Calculated: C: 58.90%; H: 8.70% Found C: 61.10%; The 13 C NMR spectrum is given in Table V. Example II. Preparation of 1 - O - (2'-propylpentanoyl) -2,3-O-isopropylidene xylitol (monoacetonexylitol valproate) and I-O- (2'-propylpentanoyl) xylitol (valproate

xylitol).

  In a thermostained reactor at 70 ° C., containing 63 ml of the dioxane water mixture, 80:20

  (v / v), 35 mL of acidic resin (AmberList 15H +) are added.

  After stirring for 2 hours under stirring, the mixture is cooled, filtered and the filtrate is evaporated under reduced pressure. The residue is taken up in ethyl ether, dried over Na 2 SO 4 and the solvent is evaporated under reduced pressure. The crude (10 g) is purified by medium pressure chromatography on silica gel (250 ml). Is successively isolated: - after passing 400 ml of hexane-ethyl ether mixture, 80:20 (v / v), 0.3 g of residual diacetonexylitol valproate; after passing 400 ml of ethyl ether 3.4 g of monoacetonexylitol valproate

(yield = 38.3%).

  [(X] = 0.01 (c = 1.90, CHCl3) Crude formula: C16H3006 Elemental analysis: Calculated: C: 60.40%, H: 9.50% Found: C 59.50%, H: 9 , 70%

"7B O

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  after passage of 400 ml of the mixture THF-ethyl ether, 50:50 (v / v), 4.6 g (yield

59.2%) of xylitol valproate.

  [?] 25 = -0.8 (c = 1.90, CHCl3) Crude formula: C13H2606 Elemental analysis: Calculated: C, 56.10%; H: 9.40% o Found C 55.60%; H: 9.50% 13C NMR spectra are given in Tables W and X.

  Example 12 Preparation of 1-0- (2'-propylpentanoyl) -2,3-isopropylidene-a-D-

  mannofuranoside (α-monoacetonemannoside valproare).

  In a reactor thermostated at 68 ° C., 35 ml of Amberlist 15H + wet acid resin and 8 g (2.1 × 10 -2 mol) of cetacetonemannoside valproate dissolved in 63 ml of

dioxane-water mixture, 80:20, v / v.

  After stirring for 30 minutes, the mixture is cooled and then filtered. The filtrate is evaporated off under reduced pressure and the residue obtained is taken up in 200 ml of ethyl ether. The solution is dried

  on anhydrous Na 2 SO 4, filtered and then evaporated under reduced pressure.

  To the 7 g of crude obtained, 100 ml of hexane are added, the mixture is brought to reflux and then filtered. The filtrate is removed and the solid phase containing the product is washed with 250 ml of hexane. The residue is dissolved in 100 ml of ethyl ether, filtered and then evaporated under pressure

scaled down.

  3.95 g (yield: 55.1%) of camonoacetonemannoside valproate are thus isolated in the form of white crystals of melting point F = 92.4 ° C.

[] 2 = 51.60 (c = 2.2, CHCl3).

  Crude formula: C 17H3007 Elemental analysis: Calculated: C: 58.90%; H: 8.70% Found C: 58.60% H: 8.70% NMR spectra 1H and 13C are given in Tables Y and Z.

  Example No. 13 Preparing 6-0- (2'-propylpentanoyl) -3-O-ocryl-1,2-O-

  isopropylidine-ca-D-glucofuranose (3-0O-ocrylmonoaceroneglucose 6-valproate).

  In a flask containing 100 ml of toluene, 6.8 g (18.10-2 mol) of valprooyl chloride and 4 ml (2.8 × 10 -2 mol) of triethylamine are added at 40 ° C. After 22 hours of reaction, the The reaction mixture is neutralized with a saturated aqueous solution of NH 4 Cl and then extracted twice with 40 ml of toluene. The organic phase is evaporated under reduced pressure. The crude (9 g) is chromatographed on silica gel with a hexane-acetone mixture, 80:20 (v / v). 4 g (yield: 65%) of 6-0-octyl-monoacetoneglucose 6valproate are thus isolated.

[c] iD2 = -31.2 (c = 1.4, CHCl3).

Gross formula C25H4607

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  Elemental analysis: Calculated: C: 65.50%; H: 10.10% Found: C: 61.00%; H: 9.80%

  Example No. 14 Preparation of 6-O- (3'-deoxy-6'-O-valproyl-1 ', 2'-O-

  isopropylidene-cz-D-glucofuranose-3'-yl) -3-0-oczyl-1,20-aD-glucofuranose isopropylidÀne-.

  In a flask containing 100 mL of toluene, 10.0 g (1.8 × 10 -6 mol) of 6-0- (3'-

  deoxy-1,2-O-isopropylidene-α-D-glucofuranose-3'-yl) -3-O-octyl-1,2-isopropylidene-α-D-

  glucofuranose, 3.6 g (2.10-2 mol) of valproyl chloride and 3.9 ml (2.8 × 10 -2 mol) of triethylamine which is heated to 40 ° C. After 22 hours of reaction, 50 ml are added. of an NH4Cl solution and then extracted with twice 40 ml of toluene. The organic phase is dried with anhydrous Na 2 SO 4 and then evaporated under reduced pressure. The crude (11 g) is chromatographed on silica gel with a hexane-acetone mixture, 80:20 (v / v). We thus isolate

  7.6 g (yield: 62%) of 6-O- (3'-deoxy-6'-O-valprooyl-1,2-Oisopropylidene-ac-D-

  glucofuranose-3'-yl) -3-O-octyl-1,2-O-isopropylidene-ca-D-glucofuranose.

F = 37-380C

[α] 2 = -31.2 (c = 1.4, CHCl 3).

  Tables A and B: NMR spectrum tH and 13C of DLACETONE GLUCOSE VALPROATE HI 5.59 d J1.2 = 3.6 Hz Cl 104.8 ppm H2 4.15 d J2.3-0 Hz C2 83.1 ppm H3 5 , 03 d J13.4 = 1.9 Hz C3 75.2 ppm H4 3, 91 dd J4.5 = 2.8 Hz C4 79.9 ppm H5 3.91 m J5.6 = 3.6 Hz C5 71.9 ppm H6 3.72 dd 16.6- = 8.5 Hz C6 67.3 ppm H6 '3.85 dd J5.6-5.6 Hz CH3 26.4 ppm CH3 1.25 s - CH3 26.4 ppm CH3 1.13 s - CH3 25, 9 ppm CH3 1.05 s - CH3 24.8 ppm CH3 1.03 s C- iso 112.9 ppm H1 "2.13 m C iso 109.0 ppm H2'- and H2 1.33 m Cl I 45, 0 ppm H3 1.13 m C2 '34.3 ppm H3' 1.13 m C2 '' 34.3 ppm H4 and H4 '' 0.64 m J = 6.5 Hz C3 20.2 ppm C3 '' 20.1 ppm C4 13, 6 ppm C4 "13.6 ppm COO 174.6 ppm

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  Tables C and D: 1H and 13C NMR spectra of DIACETONEGALACTOSE VALPROATE H1 5.25 d J1.2 = 5.0O Hz C1 95.9 ppm H2 4.04 dd J2.3 = 2.4 Hz C2 70.1 ppm H3 4.33 dd J3.4 = 7.9 Hz C3 70.4 ppm H4 3.95 dd J4 5 = 1.6 Hz C4 70.7 ppm H5 3.74 m J5.6 = 7.8 Hz C5 65.8 ppm H6 3.84 dd J6, 6- = 11.4 Hz C6 62.6 ppm H6 '4.11 dd J5.6 = 4.4 Hz CH3 25.5 ppm CH3 1.48 s - CH3 25.5 ppm CH3 1.37 s - CH3 24 , 5 ppm CH3 1.31 s - CH3 24.0 ppm CH3 1.31 s - C iso 109.1 ppm H "- 2.13 m C iso 108.2 ppm H2z and H2" 1.31 mC "44.8 ppm HY 1.13 m __C2 34.3 ppm H3 "1.13 m C2" 34.1 ppm H4a and H4 0.61 m J = 7.2 Hz C3 20.1 ppm C3 "20.1 pprn C4" 13.6 ppm C4- * 13.6 ppm COC 174.9 ppm Tables E and F: RIMN IH and 13C spectra of DIACETONEFRUCTOSE VALPROATE Hi 4.00 d | 1J.-11.5 Hz CI 64.8 ppm Hl 3.81 d C2 101, 2 ppm H3 3.96 d 13.4 = 7.8 Hz C3 70.5 ppm H4 4.33 dd J4.5 = 2.6 Hz C4 69.7 ppm H5 4.01 m J5.6 = 1.9 Hz C5 70, 1 ppm H6 3.64 dd J6.6- = 12.9 Hz C6 60.9 ppm H6 3.47 d J5.6T'0 Hz CH3 26.1 ppm CH3 1.27 s CH3 25.5 pprn CH3 1.19 s CH3 25.0 ppm CH3 1.15 s CH3 23.7 ppm CH3 1, 06 C iso 108.8 ppm H1 2.13 m C iso 108.3 ppm H2 and H2 "1.33 m C1 '44.8 ppm H3y 1.30 m C2' 34.2 ppm H3 1.06 m C2 "34, 0 ppm H4 and H4- 0.62 td I = 7.2 Hz C3" 20.2 ppm C3 "20.2 ppm C4" 13.6 ppm C4 "13.6 ppm COO 174.5 ppm Tables G and H: 1H NMR and 13C NMR spectra of the DIACEIONEXYIJIDL HI VALUE 4.15 dd J.1 = 13.5 Hz C1 63.5 ppm H1 4.03 dd I1.2 = 5.1 Hz C2 75.2 ppm H2 4 , 08 m 11J, 2 = 5.8 Hz C3 77.4 ppm H3 3.77 dd J2.3 = 7.2 Hz C4 74.9 ppm H4 4.09 dd 13.4 - = 4.4 Hz C 65, 4 ppmn H5 3.94 dd J4.5 = 6.8 Hz CH3 26.8 ppm H5 3.75 dd J5, -8.1 Hz CH3 26.7 ppm _ _ _ _ J4.5 = 8.0 HzCH3 25.9 ppm CH3 1, 30 s CH31.30sCH3 25.2 ppm CH3 1.30 s C io 1 ppm At C is 1098 ppm

CH3 1.30 s -

CH31.25 S-C iso 109.6 ppm

CH3 1.25 s -

  2Ct "145.0 ppm H1I 2.29 m3 C2 '34.4 ft H2 and H2.1.48 m = 7.8 Hz C2 34 pp C2" 34.3 ppm H3' I 1.30 m C3 '20.4 ppm Hy. 1.16 m C3. 20.4 ppm H4 'and H40.77 m J = 7.3 Hz C4' 13.8 ppm C4O 13.8 ppm COO 175.9 ppm

- '3 -

  Tables J and K: 1H NMR and 13C NMR spectra of the FDIACETONEMANNOSIDE VALPROATE 5.30 s J1.2 = 0 Hz C1 99.9 ppn H2 4.43 d J2.3 = 5.8 Hz C2 84.6 ppm H3 4 , 59 dd J3.4 = 3.5 Hz C3 79.1 ppm H4 3.69 dd J4.5 = 7.7 Hz C4 81.9 ppm H5 4.13 m J5.6-4.4 Hz C5 72 , 4 ppm H6 3.70 dd 16.6 = 8.9 Hz C6 66.6 ppm H6 3.83 dd J5.6 '= 6.3 Hz CH3 26.5 ppm CH3 1.22 s - CH3 25.6 ppm CH3 1.11 s - CH3 24.8 ppm CH3 1.11 s - CH3 24.4 ppm CH3 1.08 s - C iso 112.9 ppm H1 "2.1 m C iso 109.0 ppm H2 and H2 "C1 44.9 ppm H3" 1.04-1.31 C2 "34.2 ppm H3" C2 "34.0 ppm H4 and H4- 0.63 J = 7.2 Hz C3" 20.2 ppm C3 " 20.2 ppm C4 13.6 ppm C4 13.6 ppm COO 174.4 ppm -14 - Tables L and M: 1H NMR spectra and 13C of α-DIACETONEMANNOSIDE VALPROATE H1 5.70 s JI1.2 = 3 , 4 Hz C1 96.6 ppm H2 4.73 d J2.3 = 6.5 Hz C2 79.0 ppm H3 4.70 dd J3.4 = 3.4 Hz C3 79.4 ppm H4 3.66 dd J4 , 5 = 8.0 Hz C4 78.3 ppm H5 4.33 m J5.6 = 5.4 Hz C5 73.2 ppm H6, AAX J6.6- = 5.3 Hz C6 66.8 ppm H6 '3 , 96 J5,6 = 5.3 Hz CH3 26.9 ppm CH3 1.43 s - CH3 25.8 ppm CH3 1.32 s - CH3 25.4 ppm CH3 1.26 s - CH3 25.3 ppm CH3 1.26 s - C iso 113.9 ppm H1 I 2.32 m C iso 1090 ppm H2 'and H2CI- 45.1 ppm H3. 1.51-1.18 C2 '34.4 ppm H3 "C2-34.2 ppm H4' and H4 '0.78 J = 6.83 Hz C3' 20.4 ppm C3 '' 20.4 ppm C4 '13 , 9 ppm C4 - 13.9 ppm COO 174.0 ppmin - 1 Tables N and P: 1H and 13C NMR spectra of DIMETHYLENEXYLrOL H1 VALPROATE 4.29 decid J1,1- = 13.5 Hz Cl 62.3 ppm HI ' 4.17 dd J1'.2 = 5.1 Hz C2, 702 ppm H2 3.48 m J1.2 = 5.8 Hz C3 75.7 ppm H3 3.58 dec J2.3 = 7.2 Hz C4 69.8 ppm H4 3, 81 dd J3.4 = 4.4 Hz C5 69.2 ppm H5 4.10 dd J4.5 - = 6.8 Hz CH2 92.9 ppm H5 '3.76 dd J5.5 = 8.1 Hz CH2 92.7 ppm J4.5 '= 8.0 Hz CI' 44.9 ppm H (CH2) 5.16 s JH.H = 6.2-6A, 4 Hz C 34.4 pp C2 '34, 4 ppm H (CH2) 5.08 s JHH = 6.2-6.4 Hz C2-34.3 ppm H (CH2) 4.71 s JH.H - 6.2-6.4 Hz C 20 ppm Cy 20.3 ppm H (CH2) 4.65 s JHH = 6.2-6.4 Hz Cy. 20.3 ppm H21, 33 m c4 13.8 ppm H2 'and H2 "1, 52 m J = 7.8 Hz 138 pp C4 "- 13.8 ppm H3 1.37 m COO 176.0 ppm H3" 1.20 m H4- and H4 0.82 m J = 7.2 Hz

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  Tables Q and R: 1H NMR and 13 C NMR spectra of MONOACETONE GLUCOSE H1 5.77 d J1.2 = 3.7 Hz CI 104.8 ppm H2 4.37 d J2.3-0 Hz C2 83.1 ppm H3 5 , 17 d J3.4 = 2.5 Hz C3 76, 1 ppm H4 4.09 dd J45.3 = 8.65 Hz C4 78.9 ppm H5 no J5.6 = no C5 68.3 ppm C6 63, 9 ppm H6 3.75-3, J6.6 - Determine CH3 26.5 ppm H6. 15.6 = min. H6 - Js6'mine CH3 26.1 ppm

CH3 1.41

C iso 112.3 ppm

CH3 1.14

  1C 1- "45.1 ppm Hr '2329 C2' 34.4 ppm H2 'and H2' 1.47 H3 .... 28 C2" 34.2 ppm

H3 '1.28

  C3 "20.5 ppm H3" 1.28 ___ _ _ _ _ _C3 "20.5pprn H4 'and H4" 0.79 J = 7.2 Hz C4 "13.9 ppm C4" 13.9 ppm COO 176.4 ppm Tables S and T: 13C NMR spectra of GLUCOSE VALPROATE form IC 93.9 ppm Ci 98.7 ppm C2 72.4 ppm C2 74.6 ppmnn C3 76.7 ppm C3 78.5 ppm C4 69.9 ppm C4 69.7 ppm C5 73.5 ppm C5 78.2 ppm C6 62.4 ppm C6 623 ppmn C 1 "46.0 ppm CI" 46.0 ppm C2 '35.1 ppm C2 35.1 pprn C2 "34 , 9 ppm C2- 34.9 ppm C3 '20.7 ppm C3' 20.7 ppmN C3 '20.7 ppm C3. 20.7 ppm C4 '14.2 ppm C4' 14.2 ppm C4 '' 14.2 ppm C4 '' 14.2 ppm COO 176.3 ppm COO 176.3 ppm

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  Table U: 13 C NMR spectrum of GALACTOSE VALPROATE (PYRANOSE FORM a) Ci 93.5 ppm C2 69.9 ppm C3 70.0 ppm C4 68.3 ppm C5 70.3 ppm C6 64.0 ppm C 1 -44, 6 ppm C2 '33.9 ppm C2' 33.9 ppm C3 '19.9 ppm C3' 19.9 ppm C4 '13.8 ppm C4' 13.8 ppm COO 175.4 ppm - 19- Table V: Spectra R.MN 13C MONOACETONEFRUCTOSE VALPROATE CI 62.2 ppm C2 101.0 ppm C3 76.4 ppm C4 66.8 ppm C5 63.8 ppm C6 62.8 ppm CH3 27.4 ppm CH3 25.8 ppm C iso 1103j ppm CI "145.1ppm C2" 34.3 ppm C2 "34.3 ppm C3 20.4 ppm C3" 20.4 ppm C4 "13.8 ppm C4" 13.86 ppm COO 176.4 ppm

- 2c -

  Table W: RQ-spectrum 13C of MONOACETONEXVLITOL VALPROATE CI 63.7 ppm C2 74.9 ppm C3 77.6 ppm C4 70.4 ppm C5 63.3 ppm C 1 "44.8 ppm C2 34.1 ppm C2 34.1 ppm C3 20.1 ppm C3y 20.1 ppm C4 '13.5 ppm C4' 13.5 ppm COO 176.1 ppm 2 '- Table X: 13C NMR spectrum of XYLTOL VALPROATE CI 65.2 ppm C2 70.4 ppm C3 71.7 ppm C4 69.4 ppm C5 62.5 ppm C 1 "x4.3 ppm C2 '33.9 ppm C2"; 33.9 ppm C3 19.7 ppm C3. 19.7 ppm C4 13.6 ppm C4 "13.6 ppm COO 175.3 ppm

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  Tables Y and Z: 1H NMR and 13C NMR spectrophoton spectrophoton HL 6.09s J1.2 = 0 Hz CI 100.1 ppm H2 4.61 d J2.3 = 6.0 Hz C2 84, 6 ppm H3 4 , 84 dd 3J3.4 = 3.0 Hz C3 79.8 ppm H4 3.93 dd J4, s = nd Hz C4 81.2 ppm H5 3.93 m J5.6 = 5.4 Hz C5 69.9 ppm H6 3.57 dd J6.6- = 12.6 Hz C6 64.2 ppm OH 2.67 sl I CH3 26.0 ppm H6 '3.77 dd J5.6- = 2.59 Hz CH3 24.8 ppm CH3 1.41 C iso 113.3 ppm CH3 1.27 s Cl "45.2 ppm H1" 2.25 m 1 = 5.1 Hz C2 '34.5 ppm H2 and H2- 1.45 m C2 "34.3 ppm H3y 1.33 m C3 20.5 ppm H3 1.23 m Cy. 20.4 ppm H4 and H4- 0.77 m J = 7.2 Hz C4 '13.9 ppm C4' 13.9 ppm COO 174.9 ppm

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  Example No. 15. Toxicity and pharmacokinetics.

  1. Toxicity 1. a. Determination of LD50 Acute toxicities in SWISS mice are determined according to standard nomenclature protocols.

  Table 1. Abbreviation and numbering of molecules used in biological studies.

  Molecules abbreviation numbering valproic acid VPA I valpromide VPD II diacetoneglucose valproate VPA-DGlu II monoacetoneglucose valproate VPA-MGlu IV glucose valproate VPA-Glu Vacetonegalactose valproate VPA-DGal VI galactose valproate VPA-Gal VII dirmethylénexylitol valproate VPA- DMXyl VII diacetonexylitol valproate VPA-DXyl IX monoacetonexylitol valproate VPA-MXyl Xylitol valproate VPA-Xyl XI diacetonemannose valproate VPA-DMan XII diacetonemannose valproate VPA-DMan xIII monoacetonemannose valproate c VPA-MMan a XIV cc cc XIV diacetonefructose valproate VPA-DFru XV monoacetone valproatefructose VPA-MFru XVI monoacetoneglucose valproate-OC-8 VPA-MGlu-OC8 XVII 3-0-octyl monoacetoneglucose (6 -> 3) -6'-O- VPA-SS-OC8 XVIII valproyl of monoacetoneglucose

- 24 -

  Table 2. LD 50 values for the orle channel.

  molecule graphic value LD 50 calculated LD value 50 g / kg in mmol / kg

I 1.8 1.8 0.17 12.5

I1 1.3 1.3 + 0.21 9.09

  II> 8 n d *> 20.7 IV> 8 nd> 23.1 V> 8 nd> 26.1 VI> 8 n d> 20.7

VII 6.5 6.5 + 0.8 20.0

VIII 6.0 6.0 + 0.73 19.8

  IX> 8 n d> 22.3 XII> 8 n d> 20.7 XIII> 8 n d> 20.7 XIV> 8 nd> 14.5 XV> 8 n d> 20.7

XVI 5.0 5.0 + 0.15 14.5

  Table 3. The LD 50 value for the intraperitoneal route. molecule graphic value LD 50 calculated LD value 50 g / kg g / kg in mmol / lcg

I 0.65 0.65 + 0.08 4.50

II 0.40 0.40 0.09 2.79

III 1.80 1.80 + 0.29 4.66

IV 0.85 0.85 + 0.06 2.45

V 0.65 0.65 + 0.12 2.12

VI 1.80 1.80 + 0.36 4.66

  VIIu 1.20 1.20 0.13 3.90 Vm 3.00 3.00 + 0.28 9.93

IX 1.95 1.95 0.53 5.44

X 0.60 0.60 + 0.11 1.73

XII 1.70 1.70 + 0.23 4.40

XIII 1.70 1.70 + 0.23 4.40

XV 2.10 2.10 + 0.45 5.44

XVI 1.20 1.20 + 0.25 3.40

XVII> 4 n d> 8.73

XVIII 0.70 0.70 + 0.11 0.92

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  I. b. Pharmacological observations during acute toxicity tests Observation after ingestion or injection of increasing doses of valproic derivatives (including valpromide (If) and valproic acid (T)) shows that the animals go through 4 successive phases. : - a stirring phase; - a phase of decline in activity; - a hypnotic phase (period o can occur death);

  - an awakening and recovery phase.

  The average duration of each phase varies according to the route of administration, the nature of the

  Valprol derivative and the dose administered.

  It is noted that the action of valproic monoesters derived from monosaccharides is delayed and that the duration of the first three phases is elongated with respect to valproic acid (I) and amnide (II). These results indicate that the esters are prodrugs of the acid

  valproic since we observe the same effects with only a shift in time.

  This interpretation is confirmed by the fact that at deadly doses the death of the animal occurs

  after 6 to 10 hours for esters versus 3 to 4 hours for valproic acid.

  Table 4. Mean duration of oral (min) phase: example of glucosic esters.

  rw molecules phase I phase II phase ImI phase IV

I VPA 5 10 900 960

II VPD 5 10 900 960

  M [VPA-DGlu 10 30 900 960 IV VPA-MGIu 10 25 900 960 X V VPA-Glu T 10 20 900 960

  Table 5. Mean time of the phases (min) I.P .: example of the glucosic esters.

  r? molecules phase I phase II phase I phase IV

I VPA 2 4 900 960

  VPD 3 2 900 960 LII VPA-DGlu 10 10 900 960 IV VPA-MGIu 10 10 900 960 V VPA-Glu 10 10 900 960

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  2. Pharmacokinetic Tests in the Rat.

  These tests were carried out with the products I, II, IV and V as follows: the rats are distributed in batches of 6 after implantation of catheters. The doses used are 0.1 mmol / kg, corresponding respectively to 16.6 mg / kg for I (VPA): 38.6 mg for mI (VPA-DGlu), 34.6 mg / kg for IV (VPA- MGIu), and 30.6 mg / kg for V (VPA-Glu). The four products are administered intravenously slowly in volumes not

  not exceeding 0.1 mL for a 100g rat.

  The samples (200 μL) are made on heparin at times: 5, 15, 30, 45 minutes

* and 1, 2, 4, 6, 8, 12, 24, 48 and 72 hours.

  The blood is centrifuged (2000 rpm) for 3 minutes; 100 g of plasma are collected and stored by immediate freezing. The pellet is taken up in 100.l saline and reinjected to the animal. The concentration of circulating valproic acid is determined specifically by an enzyme immunoassay technique (EMIT test). With this assay method and for products delivered at this dose, valproic acid is detectable for 60 minutes. In addition, for the glucosic esters, the levels are very low, of the order of 1 gt / ml or in the form of traces of 2 to 4 hours depending on the case. For VPA, the concentrations are very low after 45 minutes and can not be detected later. The plasma half-life of the 3 sucroesters is 20 minutes for VPADG1u (In), minutes for VPAMAGIu (IV) and 1 hour for VPAGIu (V) respectively, versus 10 minutes for VPA.

(I).

  Example No. 16 Evidence of anriconvulsant properties in animals:

remained in penerrrazole (PTZ).

  1. Experimental protocol and results for the intraperitoneal route.

  In these experiments, we seek to trigger tonicoclonic seizures with loss of balance without causing death of the animal. In our tests, the dose of pentetrazole (> TZ) systematically induced convulsions in 97% of animals (DC 97), after

  subcutaneous injection, is 90 mg / kg.

  We use for this test male mice of 19 to 23 g (type O F I Laboratory IFFA CREDO). The animals are received at least 24 hours before the start of the tests. They are then divided into lots of 10 and each animal serves only one experiment. During studies by

  intapteroneal way animals have no access to drink or food.

  For a correct estimate of the anticonvulsant potential, it is necessary to determine the maximum peak of action of each molecule tested. The maximum peak of action is the moment o after the injection, the molecule acts most efficiently on the PZ test. The animals are distributed in batches of 10 and after intraperitoneal injection of the test compound, the mT is delivered for each lot, at a different time, at regular intervals. The number of animals with or without convulsions is then determined. Other parameters are also taken into account as the time of onset of generalized seizures and the number and intensity of signs

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  convulsive. Expression of results: the activity of a substance is expressed as an effective dose 50 (DE). This DE 50 corresponds to the theoretical dose of substance capable of suppressing maximal tonic-clonic convulsions with loss of equilibrium in 50% of the animals of the lot. It is determined graphically (logarithm-probit paper) according to the method of MILLER and

TAINTER.

  After injection of PTZ, an animal is considered to be protected if it does not show a generalized attack during a period of 30 minutes. The assessment by a criterion of all or

  nothing allows for each batch to calculate the percentage of animals with seizures.

  Drugs and solutions: the products are injected intraperitoneally in volumes not exceeding 0.20 ml for a mouse of 20 g. The solvents used are physiological saline (S (p) for water-soluble molecules, S (p) supplemented with 0.4% Tween 80 for water-insoluble liquids (S (pT) and pure olive oil (HO) for products

solid and insoluble in water.

  A solution based on DMSO (physiological serum-DMSO 90:10 (v / v)) was also used for VPD (II) and for VPA-S-S-OC8 (XVIII) as well as for the support molecules

  diacetoneglucose (DaGlu), monoacetoneglucose (MaGlu) and dimethylethylnexylitol (DMXyl).

  Valproic acid is tested in two forms; the acid form (VPA) insoluble in water

  and the sodium salt form (VPA-Na) soluble in water.

  The solubilization conditions of the molecules are described in Table 6.

  Pentetrazole is solubilized just before injection because of its poor storage in solution. It is dissolved in physiological saline at a concentration of 0.9 mg / mL, which

  corresponds to the injection of a volume of 0.20 mL subcutaneously for a mouse of 20g.

  Table 6. Solutions and concentrations of products used in the PTZ (IP) test.

  Adjuvant Products Active Ingredient mg / mL; VPA-Na concentration (1) S (p 10 - 40 VPA (I) SqpT 10-40 VPD (i) DMSO 10% 5- 20 VPA-DGIu (If) S <pT 20VPA-DGlu (III) HO 20 - 80 VPA-MGlu (IV) SqpT 20-80 VPA-Glu (V) Sp 20 80 VPA-DGal (VI) S (pT 20-80 VPA-Gal (VII) SP 20-80 VPA-DMXvI (V0) HO 20 - 80

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  VPA-DXIIIl (IX) SpT 20-80 VPA-DMan cc (X) S (pT 20-80 VPA-DMan, (XII) SpT 20-80 VPA-MGlu-OC8 (XV) SoDT 60

DM50 10%

  MaGlu-O-C8 DMOlo25 MaGlu-OC8 Tween 0.5% HO MaGIu-O-C8 S (pT 20VPA-SS-OC8 (XVIII) S (pT 50 DaGlu DMSO 10% 60 MaGlu DMSO 10% 60 DMXvl DMSO 10% 80 Results: Determination of maximum peak of action was performed for each product

(Table 7).

  Table 7. IP PTZ test Appearance of the maximum peak of action. [r Molecules Action Pic in minutes

I VPA 10

  VPA-Na 10 i, VPD 10 III VPA-DGlu (T) 30 ai VPA-DGlu (HO) 45 IV VPA-MGlu 15 V VPA-Glu 45 VI VPA-DGal 30 VII VPA-Gal 45 IX VPA-DXyl 30 The results show that all the esters have a maximum peak of action.

  delayed (15 to 45 minutes) relative to the VPA and VPD reference molecules (10 minutes).

  The values of the effective dose 50 (DE 50) at maximum peak of action are collected

in table 8.

  The valproic esters derived from saccharides have a real potential anticonvulsant vis-à-vis

  In the PTZ test, the effective doses 50 for the valproic esters ranged from 380 to 680 mg / kg, ie 1.09 mmol / kg to 1.59 mmol / kg, for the reference molecules the ED 50 are

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  respectively 150, 140 and 140 mg / kg for sodium valproate, valproic acid and valpromide. The vectors (W) delivered at high dose and at different times show no anticonvulsant action on the PIZ test, this indicates that these support molecules are not involved in the action. Valproic sucroesters significantly reduce pentetrazole-induced maximal convulsions, but in order to better evaluate their anticonvulsant potential, it is necessary to determine the protection index or the therapeutic index (ratio of effective dose 50 (ED 50) of the molecule on a toxic aspect of it, lethal dose 50 (LD 50) or

  toxic dose 50 (DT 50) for one of the tests performed).

  Table 8. IP PTZ test: Effective dose 50 measured at the peak of action.

  n Molecules graphic value DE 50 Calculated Value of 50 mg / kg. m / kg in mmol / kg I VPA-Na 150 150 + 8.2 0.9

I VPA-OH 140 140 5.1 0.95

[I VPD 65 140 4.5 0.45

  [II VPA-DGluT 450 450 27 1.11 mV VPA-DGlu HO 500 500 + 84 1.29 IV VPAMGlu 410 410 + 32 1.09 V VPA-Glu 430 430 34 1.38 VI VPA-DGal 470 470 + 19 1.23 VII VPA-Gal 380 380 + 19 1.31 VIII VPA-DMXvYl 440 440 37 1.59 IX VPA-DXyvl 480 480 83 1.21 XII VPA-DMan and 500 500 27 1.24 XII VPA-DMan f3 680 680 59 1, 42 XVII VPA-MGlu-OC8 No Ac * nd ** nd ** MaGlu-OC8 .... __

XVIII VPA-S-S-OC8. . T. "

  DaGlu MaGLU ..... DMXyi No Ac: a molecule that has not demonstrated anticonvulsant activity for this test

At * n d: not determined.

  ú5 Calculation of the protection index is commonly used in the evaluation of the anticonvulsant activity of new molecules. This criterion is in particular advocated by "the

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  Andiepileptic Drug Development Program of the Epilepsy Branch of the NIH "Aneurysm Epilepsy Development Oferogram of the US National Institutes of Health." A new antiepileptic must exert its anticonvulsant effects (ED 50) at doses

less than his TD 50.

  Table 9 gives the protection index (DE 50 / LD 50) when the test molecules are injected intraperitoneally. In this table, the reference molecule is valproate

  of sodium (activity = 1) which is compared to the activity of the esters.

  Table 9. Protection index of the molecules tested for the IP route.

  nf molecules Protection class Activity *

DL 50 / DE 50

  I VPA-Na 4.76 1 y VPD 6.25 1.31 III VPA-DGlu 4.00 0.84 IV VPA-MGIu 2.08 0.44 V VPA-Glu 1.51 0.31 VI VPA-DGal 3.85 0.81 VII VPA-Gal 3.13 0.67 VIII VPA-DMXvl 7.14 1.50 IX VPA-DXyI 4.00 0.84 XII VPA-DMan cc 2.44 0.51 XIII VPA- DMan [3 1.75 0.37 * Activity compared to that of valproic acid Of the esters tested only VPA-DMXyl has a higher protection index than VPA and even VPD for the IP route (1 , 5 against 1), the other sucroesters have a

index from 20% to 60% lower.

  If the protection index is calculated in the same way taking into account the results obtained in the neurotoxicity tests (ratio of TD 50 and ED 50), we can see, for the pathway

  [P, no molecule has a higher index than that of VPA-Na (Table 10).

  Table 10. Protection indices of the molecules tested for the IP route.

TD 50 / DE 50 TD50 / ED 50 TD 50 / DE 50

  chimney test rotorod test tensile test I VPA-Na 2.38 2.86 2.63

II VPD 0.66 0.97 0.97

III VPA-DGIu 1,11 1,88 2,63.

  IV VPA-MGlu 0.52 0.52 0.47 V VPA-Glu 1.25 1.70 1.72 VIII VPA-DMXyI 1.16 1.25 1.25

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  The esters have an index twice less than valproic acid regardless of the test chosen for the calculation of the index. The superior neurotoxicity and lower anticonvulsant properties of valproic sucroesters intraperitoneally explain this difference.

  2. Experimental protocol and results for the oral route.

  Oral assays were performed on the three glucose esters and dimethylennexylitol valproate. These tests make it possible to evaluate the duration of the anticonvulsant effect and of

  quantify the action of the product after its oral absorption.

  Mice fasted for 12 hours are weighed and divided into a batch of 6 to 10 and are force-fed the sailor between 8 and 9 hours. Food and drink are available 6 hours after force-feeding. The animals are observed the next 7 days in order to detect

  potential longer-term toxic effects.

  The drugs are administered by gastric cannulation at a dose of 10 mmol / kg. VPA-DGIu and VPA-MGlu are added with 50% olive oil. VPA-Glu is dissolved in water at 300 g / L and for VPA-DMXyl, a suspension at 12 g / L of the ester is

  carried out in a solution based on agar-agar 40g / L.

  PTZ is injected subcutaneously, in saline, every hour

for a period of 26 hours.

  The protocol for VPA is different because one can not absorb a dose equivalent to that of esters at once, its LD50 of 1.3 g / kg being too important. The animals are force-fed for this reason twice with a dose of 550 mg / kg at six o'clock

  intervals. VPA is dissolved in water at a concentration of 55 g / L.

  The results in Table 11 indicate that the maximum effect of action is delayed for sucroesters compared to valproic acid and that the duration of action is 2-3 times longer

for sucroesters.

  Table 11. Oral PTZ protection.

  molecule molecule finder Protection 80% d of the effect _________________ d __ __ __ _ total protection _ _ _ _ _ [VPA 40 minutes qq minutes 5h30 III VPA-DAGlu I hour 6 hours 16 hours [V VPA-M .IAGIu 3 hours I hour 11 hours V VPA-Glu 3 hours I hour 15 hours VIEI VPA-DMXvl 1h30 2 hours 12 hours

Example No. 15. In Vitro Tests.

  The purpose of the in vitro tests is to highlight the possible action of sucroesters - 2 -

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  before release of valproic acid.

  The study was performed on RatL hippocampal slices. Two types of epileptiform activities were induced in these slices, either by decreasing the level of calcium in the extracellular medium (Low Calcium test), or by infusing a solution containing pentetrazole, chemical convulsant previously used (PTZ test).

  l. General experimental protocol.

  The techniques described below are common to both tests:

  slices, their conservation, and the methods of registration.

  Realization of the slices: the rats are of WISTAR type male 250 to 300 g (breeding CHARLES RIVER). The animal is beheaded using a guillotine. After opening the cranial box, the hard mother is incised and folded. The brain is then extracted and immersed in a standard cooled Ringer solution saturated with carbogen (gaseous mixture 95% oxygen, 5% carbon dioxide). The hippocampus is isolated through a series of incisions after removing the overlying cortex and then placed on a receptacle. The slices are made by means of a Mc ILWAIN vertical slicer according to a cutting plane perpendicular to the major axis of the hippocampus. The hippocampal slices 450 iAm thick are ruises in a storage chamber filled with standard Ringer's solution, oxygenated continuously by bubbling of

carbogen.

  Implementation of the slices: the slices (four maximum) are placed in an electrophysiological recording chamber of the HAAS interface type. They are placed in a small chamber, the bottom of which is lined with filter paper (WHAIlTMAN No. 105) and are infused with a standard Ringer's solution. Bath temperature is maintained at 33 + 0.2

   C and the oxygenation remains constant throughout the experiment.

  Infusion solutions: a minimum recovery time of one hour, is left to the slices in the vat, infused with a standard Ringer solution; before the start of the experiments, the infusion rate is 1 mL / min. Then the different activities are induced by infusion of the solution called Low Calcium (R Ca 0) in one case and a solution containing 2 mM PTZ (R K ptz) in the other. The active ingredients to be tested are then mixed with the solutions in the form of an emulsion with Tween 80 (0.305 mM) for liquid insoluble products and in the form of a suspension with hydroxypropyl-3-cyclodextrin (HP 3 CD) 2.66 mM for solid insoluble products. These additives have been tested and they do not interfere with the electrical activities of the wafers either for Low Calcium tests or for

P'TZ tests.

  Recording: the epileptiform activity induced by the previously described solutions

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  is recorded using glass microelectrodes placed in the extracellular space. They

  are made from glass tubes (CLARK Electromedical Instruments ref GC150F-

  ), using a vertical stretching machine NARISHI [GE. They are filled with a solution of

  150 mM NaCl and connected to an amplifier and a recorder.

  The state of the slices, after the acclimation period, is evaluated by recording responses to electrical stimulation delivered by a bipolar tungsten semi-microelectrode at afferents (collaterals of Schaffer) that form synapses on neurons.

  recorded. Slices are in good condition if they give a quick and ample answer.

  Records of extracellular field potentials are obtained in the CA I region of the hippocampus (area n0 I Amon's Horn) at the level of the cell layer

  pyramid. The slices are then perfused by the solutions triggering the activities.

  2. Low Calcium Experiment When calcium is omitted from the perfusion solutions delivered, epileptiform spontaneous activities develop in the absence of synaptic transmission after one hour. These activities appear in the area CA 1, this recurrently with a frequency

  stable. They are characterized by a negative potential during extracellular recording.

  Procedure: After infusing the slices with a Low Calcium solution for an average period of 45 minutes, spontaneous epileptiform

  of varied intensities and frequencies.

  The products to be studied solubilized in the R Ca 0, are mixed with the infusion which is maintained until the activities cease or, in the other cases, during a period

maximum of 20 minutes.

  The slices are then again infused in Low Calcium until the return of the epileptiform activity. This washing is observed for a shorter or shorter period, depending on the

products and their concentration.

  Each of the substances was tested at different concentrations; for each of

  concentrations, we used at least 5 slices.

  Analysis of the data: the paroxysmal activities are magnified 1000 times, recorded in the frequency zones located between 10 Hz and 10 KHz and followed on an oscilloscope. Signals are digitized (CED 1401 scanning interface) and stored in memory

  computer. The SPIKE 2 program analyzes the data and allows deferred processing.

  For all the molecules tested, the minimum inhibitory concentration is determined and

  the total suppression time of the activity is recorded.

  Drugs and solurions: four valproic esters were tested: diacetoneglucose valproate (VPA-DGlu), monoacetoneglucose valproate (VPA-MGlu), valproate 34-

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  Glucose (VPA-GIu) and dimethylenexylitol valproate (VPA-DMXyl) as well as four standard antiepileptic drugs, sodium valproate (VPA-Na), valpromide (VPD), phenobarbital (PILE) and phenytoin (DPH). ). The solubilization conditions are described in

the tables below.

  VPA-DGIu (Em) Molecules VPA-.MGIGu (IV) VPA-Glu (V) VPA-DMXyl (VIi) Concentration 1 to 5 MM 1 to 5 rm 1 to 5 M0, 1 to 1 mmn Solution R Cao R Cao R Ca (R) Ca (Additive Tween Tween HP J CD HP D CD Molecules VPA-Na (I) VPD (II) PHE DPH Concentration I at 5 mI.M l at 5 mM 0, l at m, II mM Solution R Cao R Cao R Cao R Ca (Additive ... HP _ CD

  Choice of the animal: the rats are of type Wistar whose weight is lower than 300 g.

  Influence of the potassium concentration: the optimal potassium concentration has been previously determined and is between 6 and 7 mM. The value for our experiments was set at

6.5 mM.

  Results: after the introduction of the active molecule, there is a gradual decrease in the amplitude of the field potentials that can go to their complete disappearance. The minimum effective doses (DEM) capable of completely inhibiting in less than 20 minutes the low calcium activities are: 1 mM for glucose valproate (V): 1 mM for monoacetone glucose valproate (IV); - 0.25 mM for diacetone glucose valproate (II);

  0.1 mM for dimethylennexylitol valproate (VI).

  reference antiepileptics with an EMR of: -> 10 mM for VPA (I): - 2.5 mM for VPD (IU); - I MNM for phenobarbital:

- 0.25mam for phenitoine.

3. Experiences with the PTZ.

  The action of PTZ in vitro is studied on the rat hippocampus because it induces discharges

  paroxysmal. This test allows the evaluation of the activity of anticonvulsant molecules.

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  Operating mode: the protocol for setting up and registering the slices is the same as that described previously. Slices are infused with R K PTZ solution. The paroxysmal spontaneous discharges appear in 20 minutes on average. After a period of time to stabilize the activity, the drugs are infused. The products are applied for 20 to 60 minutes depending on the doses, then the slices are again infused with the R K PTZ until a total return of the initial activity. The tests are at each

  repeated on at least four different slices.

  Drugs and solutions. the substances to be studied are then applied and the effect on the frequency of activities analyzed. Hydroxy-3cyclodextrin (HP 1 CD) is used at the

concentration of 2.66 mM.

  VPA-Na (I) VPD (II) VPA-Glu (V) VPA-DMXyl (VE) Molecules Concentration 2 mM 1 and 2 rmv 1 and 2 rnM 0.1 and 0.5 rnmM RK solution PTZ RK pTZ RK Pz RK ETZ Additive --- .. HP CD HP C CD Data analysis. The burst discharges are amplified 1000 times and recorded in the frequency zones located between 10 Hz and 10 KHz and observed on an oscilloscope. Signals are recorded on a paper recorder (GOULD) with bandwidth

reduced (10-200Hz).

  The evolution of the number of discharges per time interval is noted dug all the test.

  The values are then transcribed in a table and translated graphically to obtain the

final curves.

  Results: The in-vitro tests show that the suppression of the activities induced by PTZ is obtained at the dose of: -0.1 μm for VPA-dimethyvinylnexylitol (VIII) (95% inhibition after 40 min of infusion); 0.5 mM for VPA-dimethylennexylitol (VIII) (100% inhibition after 20 min of infusion); -2 mMl for VPA-diacetoneglucose (III) (100% inhibition after 34 min infusion); 2 mM for VPD (II) (100% inhibition after 12 min infusion); At 2 mM there is no inhibition by sodium valproate (I) (the percentage

  inhibition is less than 50%).

  In all cases, the activity induced by PTZ is found after stopping the infusion of the

pnrinclpe active.

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Claims (14)

claims   1. Compounds corresponding to the general formula: W- (OCO-CH (CH 2 CH 2 CH 3) 2) n in which W represents a radical of a product having at least one OH group esterifiable by valproic acid or its derivatives, or Y leaving part may be substituted by the valproate group, n may vary from I to the total number of esterifiable OH groups or Y substitutable in said product. The ester or polyester compounds of the present invention have a radical W having for its precursor W-OH which may be a monosaccharide, a polysaccharide or an itol, functionalized or otherwise, protected or not, and preferably chosen from D-glucose.   or its derivatives 1,2,5,6-diaceconeglucose or 1,2: 3,5-diacetoneglucose or 1,2-   monoacetoneglucose, D-galactose or diacetonegalactose, Dfructose or its derivatives   2,3,4,5-diacetonefructose or 1,2,4,5-diacetonefructose or 1,2-monoacetonefructose or 2,3-diacetonefructose   monoacetonefructose, D-mannose or its derivatives 2,3: 5,6-diacetonemannose or 2,3-   monoacetonemannose, D-allose or its derivatives 1,2: 5,6-diacetonealllose or 1,2-   monoacetone, D-xylose or 1,2-monoacetonexylose, xylitol or its derivatives 2,3: 4,5-   diacetonexylitol or 2,3-monoacetonexylitol or 4,5-monoacetonexylitol or 2,4: 3,5-   dimethylènexylitol or from disaccharides and polysaccharides that may result from the combination of two or more of the monosaccharides previously mentioned. The precursor is glycerol and soalketal. Compoundable compounds are excluded valproic esters corresponding to the above general formula in which W represents the unit   glycerol or solketal, 1,3-dipalmitoyl-2-valproyl-glycerol, 1,3-didecanoyl-2-valproyl-   glycerol, 1,3-dihexanoyl-2-valproyl-glivcerol, 1,3-dibutanoyl-2valproyl-glycerol, 1,3-   diacetyl-2-valproyl-glycerol, 2-valproyl-glycerol, 1,3-divalproylglycerol, 2-acetyl-1,3-   divalproyl-glycerol, 2-butanoyl-1,3-divalproyl-glycerol, 2-butanoyl-1I, 3-divalproyl-   glycerol, 1,3-divalproyl-glycerol.   2. 3-O-valproyl-D-glucose and its derivatives 3-O-valproyl-1,2: 5,6-di-O-isopropylidene   t-D-glucofuranose and 3-O-valproyl-1,2-O-isopropylidene-α-D-glucuranose.   3. 6-O-Valproyl-D-galactose and the 6-O-valproyl-1,2: 3,4-di-O-isopropylidene derivative .alpha.-D-galactopyranose.   4. 1-O-valproyl-D-mannoside and its derivatives 1-O-valproyl-2,3-Oisopropylidene-D-   mannofuranoside and I-O-valproyl-2,3: 5,6-di-O-isopropylidene-Dmannofiranoside.   5. 1-O-valproyl-5-D-frucrose and its derivatives 1-O-valproyl-2,3-isopropylidene-p-D-   fructopyranose and 1-O-valproyl-2,3 :: 4,5-di-O-isopropylidene-5-dfructopyranose.   6. 3-O-valprovl-5-D-fructose and its derivatives 3-O-valproyl-1,2-isopropylidene-3-D-   fiructopyranose and 3-O-valproyl-1,2,4,5-di-O-isopropylidene-1-D-fructopyranose.   7. 1-O-valprovl-xylitol and its derivatives 1-O-valproyl-2,4-isopropylidene-xylitol, 1- - ORIGINAL BA BAD ORIGINAL   O-Valproyl-3,5-O-Isopropylidene-xylitol and 1-O-Valproyl-2,4: 3,5di-Oisopopylid ne-xylitol   8. 1-O-Valproyl-2,3,4,5-di-O-methylen-xylitol.   9. 6-O-Valproyl-3-O-octyl-D-glucose and 6-O-valproyl-3-O-octyl-1,2-O- isopropylidene-α-D-glucofuranose.   10. 6-0- (3'-Deoxv-6'-O-valproyl) -1 ', 2'-O-isopropylidene-α-D-Glucofuranose-3'-   yivl) -3-O-octyl-1 '2-O-isopropylidene-α-D-glucofuranose.   11. Relaxing medicaments comprising the valproic esters according to any one of   Claims I to 10, used alone or in combination with other compounds.   12. Hypnotic medicinal products comprising valproic esters, whichever   Claims I to 10 used alone or in combination with other compounds.   13. Anticonvulsant medicaments comprising the empiric esters according to one of   any of claims I to 10, used alone or in combination with other compounds.   14. Antiepileptic drugs comprising valproic esters according to one of   any of claims I to 10, used alone or in combination with other compounds. BAD ORIGINAL