EP0352301A1 - Esters of n-butyric acid and their use as drugs - Google Patents

Esters of n-butyric acid and their use as drugs

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Publication number
EP0352301A1
EP0352301A1 EP88908255A EP88908255A EP0352301A1 EP 0352301 A1 EP0352301 A1 EP 0352301A1 EP 88908255 A EP88908255 A EP 88908255A EP 88908255 A EP88908255 A EP 88908255A EP 0352301 A1 EP0352301 A1 EP 0352301A1
Authority
EP
European Patent Office
Prior art keywords
butanoyl
isopropylidene
compound
butyric acid
compound according
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP88908255A
Other languages
German (de)
French (fr)
Inventor
Pierre Joseph Villa
François PIERI
Gino Lino Ronco
Emile Pierre René SEGARD
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Universite de Picardie Jules Verne
Original Assignee
Universite de Picardie Jules Verne
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from FR8713294A external-priority patent/FR2621039B1/en
Application filed by Universite de Picardie Jules Verne filed Critical Universite de Picardie Jules Verne
Publication of EP0352301A1 publication Critical patent/EP0352301A1/en
Withdrawn legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H13/00—Compounds containing saccharide radicals esterified by carbonic acid or derivatives thereof, or by organic acids, e.g. phosphonic acids
    • C07H13/02—Compounds containing saccharide radicals esterified by carbonic acid or derivatives thereof, or by organic acids, e.g. phosphonic acids by carboxylic acids
    • C07H13/04—Compounds containing saccharide radicals esterified by carbonic acid or derivatives thereof, or by organic acids, e.g. phosphonic acids by carboxylic acids having the esterifying carboxyl radicals attached to acyclic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D317/00—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms
    • C07D317/08—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms having the hetero atoms in positions 1 and 3
    • C07D317/10—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms having the hetero atoms in positions 1 and 3 not condensed with other rings
    • C07D317/14—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms having the hetero atoms in positions 1 and 3 not condensed with other rings with substituted hydrocarbon radicals attached to ring carbon atoms
    • C07D317/18—Radicals substituted by singly bound oxygen or sulfur atoms
    • C07D317/24—Radicals substituted by singly bound oxygen or sulfur atoms esterified

Definitions

  • Esters of n-butyric acid and their use as medicaments are also useful as medicaments.
  • the present invention relates to derivatives of n-butyric acid and their use as a medicament.
  • the salts of n-butyric acid, and in particular sodium butyrate have interesting inhibitory actions on the growth of fibroblasts as a result of actions on histones and the structure of chromatin in cell nuclei.
  • This inhibitory action of cell division has proved particularly important to exploit in cancerology, in particular for the treatment of various leukemias or sarcomas and various carcinomas.
  • antiviral actions of sodium butyrate have also been characterized for the herpes virus.
  • n-arginine butyrate gives the following results: 2 min: 1.36 mmol / l; 5 min: 0.23 mmol / l; 30 min: 0.06 mmol / l; 60 min: 0 mmol / l.
  • the Applicant has set itself the goal of synthesizing chemical compounds containing in their structure at least one butanoyl radical linked by a covalent bond to an atoxic carrier compound so that these compounds in vivo under the action of enzyme systems in humans or animals slowly release n-butyric acid, resulting in a longer half-life than that of the above saline derivatives and therefore providing better bioavailability of the biologically active product.
  • W represents a radical of a product having at least one OH group esterifiable by n-butyric acid, n possibly varying from 1 to the total number of esterifiable OH groups in said product.
  • the product, of radical W plays the role of a temporary transporter of n-butyric acid and it is particularly advantageous to choose it from products which are the least toxic possible with respect to organisms. alive, to the extent that this product is released into the body and must therefore be eliminated or metabolized.
  • the ester compounds of the present invention will have a radical W forming part of products chosen from carbohydrates and their acetonide derivatives, hydrogenated carbohydrates and their acetonide derivatives, glyceryl esters of fatty acids, the derivative glycerol acetonide, serine and homoserine.
  • the compounds according to the present invention are intended for use as a medicament alone or in combination with other compounds, intended for use as n-butyric acid and its salts, for example for anti-tumor and / or antiviral use.
  • the collected filtrate is evaporated under reduced pressure.
  • the product obtained (31.69 g) is a yellow pasty liquid with rotary power
  • [ ⁇ ] D 22 - 28o (ethanol).
  • the CPV analysis (OV 17 ; 1 m; 200o C; 1.7 bar; retention time 3 min) indicates a purity of the order of 98%.
  • the entire product is then chromatographed on silica (Kielselgel 60, 150 g) using a heptane / ethyl ether gradient (fraction 90/20 v / v).
  • the mono-isopropylidene derivative is prepared from the di-isopropylidene derivative (compound I) prepared under the conditions of Example 1.
  • the vapor phase chromatography shows the disappearance of the butyrate-3 peak of di-isopropylidene glucose and the solution is neutralized with tripropylamine, the sulfate of which is insoluble in the medium.
  • the solvent is removed under reduced pressure.
  • the crude fraction obtained is purified by liquid chromatography on silica (Kielselgel 60 Merck 9585, 230 - 400 mesh, 150 g) using a heptane / acetone gradient (80/20 v / v fraction).
  • This quantity can be increased when the above acid hydrolysis is carried out at the higher temperature of 80oC and by increasing the contact time (3 hours).
  • This compound is prepared by isomerization of O-butanoyl-3 O-isopropylidene-1,2 ⁇ -D-glucofuranose (II) synthesized under the conditions of Example 2. It is dissolved. 30 g of II in 1 liter of methanol containing 10 ml of triethylamine; the mixture is stirred at room temperature for 2 hours. After evaporation under reduced pressure, the syrup obtained, fractionated by preparative HPLC on a WATERS Prep column. pack P / N 500-41.75 mesh (eluent hexane-acetone, 80/20 v / v), gives:
  • This compound is prepared from di-O-isopropylidene-1,2: 3,4 ⁇ -D galactopyrannose according to the method of Example 1 with a yield of 62% after recrystallization from hexane.
  • This compound is prepared from VIII by deprotection of the hydroxyls in 2 and 3, ie in the homogeneous phase according to the method of Example 2 slightly modified (6 hours at room temperature instead of 1 hour at 70 ° C); either in the heterogeneous phase by passage through a column of 25 ml of Amberlist resin. 15 Wet maintained at 30oC with a flow rate of 0.1 ml / min of VIII in 20% solution in an isopropanol-water mixture 90-10 v / v. The purification is carried out on a column of silica gel with ethyl ether as eluent.
  • This compound is prepared from compound IX according to the method of Example 1 with a yield of 72% after purification on silica gel.
  • This compound is prepared according to the following sequence: a) esterification with palmitoyl chloride of
  • Example 1 leading to O-palmitoyl-1 O-isopropylidene-2,3 propane; b) deprotection of the hydroxyls in 2 and 3 of the compound obtained in a) according to the methods described for the preparation of IX, leading to O-palmitoyl-1 propane diol-2,3; c) esterification with butanoyl chloride of the compound obtained in b) according to the method of Example 1 leading to XI, with a yield of 70% after purification on silica gel. Liquid, n D 20 1.4475.
  • a solution containing 0.600 g of the compound according to the invention is prepared in 2 ml of formal glycerol (mixture of dioxane - 1.3 ol-5 and dioxolane - 1.3 methanol-4).
  • formal glycerol mixture of dioxane - 1.3 ol-5 and dioxolane - 1.3 methanol-4.
  • a prior injection 0.1 ml / kg
  • a heparin solution at a concentration of 5000 IU / ml is carried out in the marginal vein of the ear; the needle is left in place allowing administration, two minutes later, of a dose of 0.1 ml / kg (i.e. 23 mg / kg) of the solution prepared above.
  • Blood samples of 1 cm3 spread over time are taken from the marginal vein of the other ear. The blood is centrifuged (2,000 rpm for 3 min) and the serum is collected.
  • the concentration of potential serum butyric acid is determined by gas chromatography using a column of CHROMOSORB 101 or CARBOPACK C.
  • the sample to be analyzed by chromatography is prepared by the following treatment: - 80 ⁇ l of serum - 10 ⁇ l of pure formic acid
  • This treatment before chromatography has the effect of hydrolyzing the compound or its metabolites and releasing n-butyric acid.
  • the quantification is done by comparison of the respective surfaces of butyric acid and valeric acid, the response coefficients having been determined beforehand.
  • the serum concentration (in mmoles) of potential or releasable n-butyric acid is thus measured from the compound or its metabolites contained in the serum. These measurements make it possible to determine the apparent serum half-life of the injected ester compound.
  • the compounds according to the invention have a duration of
  • mice 3o In-vivo anti-tumor tests in mice.
  • the anti-tumor properties studies were carried out in mice, according to the protocol of CHANY and CERUTTI (Int. J. Cancer, 1982, 30, 489) on the compounds I to V above.
  • the TG 180 strain is used (ip injection of 10 6 cells for a 25 g male mouse and 0.1 ml of a solution of Corynebacterium Parvum Mérieux (CP)).
  • CP Corynebacterium Parvum Mérieux
  • mice 9 lots of 15 mice each. After injection on day D O of TG 180 cells and of CP, the procedure is as follows: The control batch No. 1 is. treated daily with a placebo injection. Control batch 2 is treated daily with an injection of CP (0.1 ml).
  • Control batch 4 is treated daily with CP and then interferon at the same doses as above.
  • the experimental batches are treated, alternately on D + 1 with the compound according to the invention (0.5 ml of an 8 mmolar solution in compound I to V), on D + 2 with interferon, on D + 3 by CP and this for three weeks.
  • Table 4 shows the very good therapeutic anti-tumor efficacy of the compounds I, II, III according to the invention, compared with the control batches 1, 2, 3 and 4.
  • the amount of potential n-butyric acid which can be released in the mouse body is in this protocol for compounds I, II, III of 14 mg per kilogram of mouse, which corresponds in theory to the optimal dose recommended for arginine butyrate for a 25 g mouse. Second experience.
  • Tests according to the same protocol as above to determine the dose-effect of the compounds according to the invention were carried out by keeping constant the amount of CP and of interferon from the previous experiment, but by varying the concentration of 0.25 to 50 millimoles of compound II per liter of solution for injection (0.5 ml of this solution being injected), ie the potential equivalent of 0.44 to 88 mg of n-butyric acid per kg of mouse.
  • the survival at 100 days is 1 mouse out of 45 for CP alone, 0 out of 45 for interferon alone, 30 out of 45 for compound II, whatever its concentration between 0.25 and 50 mmol / l, the compound II being administered alternately with CP and interferon according to the above protocol.
  • Another trial according to another protocol in which the interferon has been suppressed but in which one continues to administer alternately 0.1 ml of CP and 0.5 ml of a solution of compound II at a concentration varying from 0, 25 to 50 millimoles / l, shows that the number of mice surviving to 100 days is constant whatever the concentration of compound II and amounts on average to 20 out of 45 mice.
  • compositions are prepared for therapeutic use in humans or animals comprising 100 to 2000 mg of a compound according to the invention and any pharmaceutically acceptable excipient.
  • compound II is placed in isotonic solution with physiological serum or glucose to form an injectable drug dosed unitarily between 100 to 500 mg of compound II.
  • Compound II is optionally mixed with an excipient such as a polyethylene glycol (PEG) to form a medicament which can be administered orally or rectally and dosed individually between 1 and 2 g of compound II.
  • PEG polyethylene glycol
  • gastro-resistant capsules are provided to avoid acid hydrolysis in the stomach.

Abstract

Alors que les butyrates de sodium et d'arginine ont une durée de demi-vie inférieure à cinq minutes, dans le courant sanguin du lapin, à la suite d'une injection i.v, les esters de l'acide n-butyriques ont une durée de demi-vie beaucoup plus longue. Ils constituent ainsi une réserve potentielle en acide butyrique par hydrolyse enzymatique lente in-vivo. Des essais pharmacologiques sur des souris porteuses de tumeurs montrent que le dosage thérapeutique en mole de ces esters, par rapport aux butyrates salins, peut-être réduit d'environ 30 fois. Application de ces esters pour l'obtention de médicaments avec peu d'effets secondaires et destinés à une utilisation antitumorale et/ou antivirale.Whereas sodium and arginine butyrates have a half-life of less than five minutes, in the rabbit blood stream following iv injection, n-butyric acid esters have a half-life of less than five minutes. much longer half-life. They thus constitute a potential reserve of butyric acid by slow in-vivo enzymatic hydrolysis. Pharmacological tests on tumor-bearing mice show that the therapeutic dosage in moles of these esters, compared to saline butyrates, can be reduced by about 30 times. Application of these esters for obtaining drugs with few side effects and intended for antitumor and/or antiviral use.

Description

Esters de l'acide n-butyrique et leur utilisation comme médicaments. Esters of n-butyric acid and their use as medicaments.
La présente invention concerne des dérivés de l'acide n-butyrique et leur utilisation comme médicament. Il est connu que les sels de l'acide n-butyrique, et tout particulièrement le butyrate de sodium, possèdent des actions inhibitrices intéressantes sur la croissance des fibroblastes par suite d'actions sur les histones et la structure de la chromatine des noyaux cellulaires. Cette action inhibitrice de la division cellulaire s'est révélée particulièrement importante à exploiter en cancérologie, notamment pour le traitement de diverses leucémies ou de sarcomes et carcinomes divers. Outre ces actions antimitotiques au niveau des noyaux cellulaires, des actions antivirales du butyrate de sodium ont été également caractérisées pour le virus de l'herpès. Cependant, il est connu également à la suite des tests pharmacocinétiques que les sels minéraux de l'acide n-butyrique ont une durée de vie in vivo qui est extrêmement brève. Pour tenter d'accroître la demi-vie du produit injecté, on a proposé par ailleurs d'autres dérivés comme par exemple, le butyrate d'arginine (Ch. CHANY et coll., Int.J. Cancer, 1982, 30, 489; ibid. 1983, 32, 379 et Demande de brevet. EP-A1-0 069 659 (INSERM)). Ces dérivés ioniques ne permettent pas d'obtenir une durée de vie in vivo suffisante, pour une action pharmacologique satisfaisante, car ils sont éliminés très rapidement après leur administration par voie intravéneuse. Ainsi si du n-butyrate de sodium est injecté au temps t = 0 par voie iv à raison de 0,12 g par kilogramme de lapin, les dosages de l'acide n-butyrïque présent en mmol par litre de sérum sont au temps t = 2 min: de 1,66 mmol/l: 5 min : 0,87 mmol/l ; 18 min : 0,03 mmol/l; 30 min : 0,02 mmol/l.The present invention relates to derivatives of n-butyric acid and their use as a medicament. It is known that the salts of n-butyric acid, and in particular sodium butyrate, have interesting inhibitory actions on the growth of fibroblasts as a result of actions on histones and the structure of chromatin in cell nuclei. This inhibitory action of cell division has proved particularly important to exploit in cancerology, in particular for the treatment of various leukemias or sarcomas and various carcinomas. In addition to these antimitotic actions at the level of cell nuclei, antiviral actions of sodium butyrate have also been characterized for the herpes virus. However, it is also known from pharmacokinetic tests that the mineral salts of n-butyric acid have an extremely short lifespan in vivo. In an attempt to increase the half-life of the injected product, other derivatives have also been proposed such as, for example, arginine butyrate (Ch. CHANY et al., Int.J. Cancer, 1982, 30, 489 ; ibid. 1983, 32, 379 and Patent application. EP-A1-0 069 659 (INSERM)). These ionic derivatives do not make it possible to obtain a sufficient in vivo lifetime for a satisfactory pharmacological action, since they are eliminated very quickly after their administration by the intravenous route. Thus if sodium n-butyrate is injected at time t = 0 by the iv route at a rate of 0.12 g per kilogram of rabbit, the dosages of n-butyric acid present in mmol per liter of serum are at time t = 2 min: from 1.66 mmol / l: 5 min: 0.87 mmol / l; 18 min: 0.03 mmol / l; 30 min: 0.02 mmol / l.
Dans les mêmes conditions le n - butyrate d'arginine donne les résultats suivants : 2 min : 1, 36 mmol/l ; 5 min: 0,23 mmol/l ; 30 min : 0,06 mmol/l ; 60 min : 0 mmol/l.Under the same conditions, n-arginine butyrate gives the following results: 2 min: 1.36 mmol / l; 5 min: 0.23 mmol / l; 30 min: 0.06 mmol / l; 60 min: 0 mmol / l.
Ces résultats pharmacocinétiques montrent que la demi-vie de ces composés dans le courant sanguin du lapin est très brève et inférieure à 5 min. En partant des conclusions actuellement, admises à partir des études réalisées, il est admis que, pour les n-butyrates salins ci-dessus, le produit actif biologiquement est l'acide n-butyrique.These pharmacokinetic results show that the half-life of these compounds in the rabbit's bloodstream is very brief and less than 5 min. Starting from the conclusions currently accepted from the studies carried out, it is accepted that, for the saline n-butyrates above, the biologically active product is n-butyric acid.
La Demanderesse s'est fixée comme but de synthétiser des composés chimiques contenant dans leur structure au moins un radical butanoyle lié par une liaison covalente à un composé porteur atoxique de façon à ce que ces composés in-vivo sous l'action des systèmes enzymatiques chez l'homme ou l'animal libèrent lentement de l'acide n-butyrique avec comme résultat une durée de demi-vie plus longue que celle des dérivés salins ci-dessus et procurant donc une meilleure biodisponibilité du produit biologiquement actif.The Applicant has set itself the goal of synthesizing chemical compounds containing in their structure at least one butanoyl radical linked by a covalent bond to an atoxic carrier compound so that these compounds in vivo under the action of enzyme systems in humans or animals slowly release n-butyric acid, resulting in a longer half-life than that of the above saline derivatives and therefore providing better bioavailability of the biologically active product.
Ce but est atteint par les composés selon 1 '-invention de formule : W-(O-CO-CH2-CH2-CH3)n dans laquelle W représente un radical d'un produit ayant au moins un groupe OH estérifiable par l'acide n-butyrique, n pouvant varier de 1 au nombre total des groupes OH estérifiables dans ledit produit. Dans le cadre de la présente invention le produit, de radical W, joue le rôle d'un transporteur temporaire d'acide n-butyrique et il est particulièrement avantageux de le choisir parmi des produits les moins toxiques possibles vis-à-vis des organismes vivants, dans la mesure où ce produit est libéré dans l'organisme et doit être par conséquent éliminé ou métabolisé.This object is achieved by the compounds according to the invention of formula: W- (O-CO-CH 2 -CH 2 -CH 3 ) n in which W represents a radical of a product having at least one OH group esterifiable by n-butyric acid, n possibly varying from 1 to the total number of esterifiable OH groups in said product. In the context of the present invention, the product, of radical W, plays the role of a temporary transporter of n-butyric acid and it is particularly advantageous to choose it from products which are the least toxic possible with respect to organisms. alive, to the extent that this product is released into the body and must therefore be eliminated or metabolized.
D'une manière préférée les composés ester (s) de la présente invention auront un radical W faisant parti de produits choisis parmi les carbohydrates et leurs dérivés acétonides, les carbohydrates hydrogénés et leurs dérivés acétonides, les esters glycéryliques d'acides gras, le dérivé acétonide du glycérol, la serine et l'homosérine. Ces radicaux W préférés sont métabolisés en composés assimilables par l'organisme avec éventuellement la formation in-situ d'acétone, composé peu toxique (DL50p.o. = 10,7 ml/kg de rat - Merck Index 10ième édition). Les composés dérivés de monosaccharide aldose ou cétose, cyclique ou acyclique, et leurs acétonides sont encore plus préférés car l'hydrolyse in-vivo libère avec éventuellement de l'acétone, de l'acide butyrique et un monosaccharide, élément nutritif courant de l'homme ou de l'animal, donc atoxique.Preferably, the ester compounds of the present invention will have a radical W forming part of products chosen from carbohydrates and their acetonide derivatives, hydrogenated carbohydrates and their acetonide derivatives, glyceryl esters of fatty acids, the derivative glycerol acetonide, serine and homoserine. These preferred W radicals are metabolized into compounds which can be assimilated by the body, possibly with the in situ formation of acetone, a slightly toxic compound (LD 50 in = 10.7 ml / kg of rat - Merck Index 10th edition). Compounds derived from cyclic or acyclic aldose or ketosis monosaccharide and their acetonides are even more preferred because the in-vivo hydrolysis releases with possibly acetone, butyric acid and a monosaccharide, a common nutrient of the man or animal, therefore non-toxic.
Les composés selon la présente invention sont prévus pour une utilisation comme médicament seul où en association avec d'autres composés, destiné(s) comme l'acide n-butyrique et ses sels, par exemple à une utilisation antitumorale et/ou antivirale.The compounds according to the present invention are intended for use as a medicament alone or in combination with other compounds, intended for use as n-butyric acid and its salts, for example for anti-tumor and / or antiviral use.
Outre les dispositions qui précèdent, l'invention comprend encore d'autres dispositions, qui ressortiront de là description qui va suivre. Il doit être bien entendu, toutefois, que cette description et les exemples qu'elle contient sont donnés uniquement à titre d'illustration de l'objet de l'invention dont ils ne constituent en aucune manière une limitation. Exemple 1.In addition to the foregoing provisions, the invention also comprises other provisions, which will emerge from the description which follows. It should be understood, however, that this description and the examples it contains are given only by way of illustration of the subject of the invention of which they do not in any way constitute a limitation. Example 1.
Préparation du O-butanoyl-3-di-O-isopropylidène 1,2 : 5,6α-D-glucofurannose (I).Preparation of O-butanoyl-3-di-O-isopropylidene 1,2: 5,6α-D-glucofuranose (I).
Dans un flacon tricol, on place 26 g (0,1 mole) de di-O-isopropylidène-1,2 : 5,6α-D-glucofurannose préparé par ailleurs selon les procédés connus. On introduit 100 ml d'heptane, puis 19 ml (0,1 mole) de tripropylamine. On chauffe l'ensemble sous agitation jusqu'à reflux et on ajoute alors goutte à goutte 10 ml de chlorure de butanoyle (0,1 mole) en solution dans le heptane (50 ml).In a three-necked flask, 26 g (0.1 mole) of di-O-isopropylidene-1,2: 5,6α-D-glucofuranose prepared in addition according to known methods are placed. 100 ml of heptane are introduced, then 19 ml (0.1 mole) of tripropylamine. The whole is heated with stirring until reflux and then 10 ml of butanoyl chloride (0.1 mol) in solution in heptane (50 ml) are added dropwise.
On poursuit le reflux pendant 60 mn après la fin de l'addition. Après retour du mélange réactionnel à la température ambiante, ce dernier est filtré afin d'éliminer les cristaux de chlorhydrate de tripropylamine formés et les restes éventuels de di-O-isopropylidène 1,2 : 5 , 6 α -D-glucofurannose.The reflux is continued for 60 min after the end of the addition. After returning the reaction mixture to room temperature, the latter is filtered in order to remove the crystals of tripropylamine hydrochloride formed and the residues di-O-isopropylidene 1,2: 5, 6 α -D-glucofuranose.
Le filtrat recueilli est évaporé sous pression réduite. Le produit obtenu (31,69 g) est un liquide pâteux jaune de pouvoir rotatoireThe collected filtrate is evaporated under reduced pressure. The product obtained (31.69 g) is a yellow pasty liquid with rotary power
[α]D 22 = - 28º (éthanol). L'analyse C.P.V. (OV17;1 m ; 200º C ; 1,7 bar ; temps de rétention 3 mn) indique une pureté de l'ordre de 98 % . La totalité du produit est ensuite chromatographiée sur silice (Kielselgel 60, 150 g) à l'aide d'un gradient heptane/ether éthylique (fraction 90/20 v/v).[α] D 22 = - 28º (ethanol). The CPV analysis (OV 17 ; 1 m; 200º C; 1.7 bar; retention time 3 min) indicates a purity of the order of 98%. The entire product is then chromatographed on silica (Kielselgel 60, 150 g) using a heptane / ethyl ether gradient (fraction 90/20 v / v).
Le produit purifié (26,75 g) se présente sous la forme d'un liquide visqueux jaune très pâle de pouvoir rotatoire [α]D 22 = - 32 , 67º (éthanol), d'indice de réfraction n = 1,477 à 23º C, et peu soluble dans l'eau.The purified product (26.75 g) is in the form of a very pale yellow viscous liquid with rotary power [α] D 22 = - 32, 67º (ethanol), with a refractive index n = 1.477 at 23º C , and sparingly soluble in water.
Le rendement global est de 81 % en produit ainsi purifié. Les spectres RMN 1H et 13C sont donnés en annexe sur les tableaux 1 et 2. Exemple 2 :The overall yield is 81% of product thus purified. The 1 H and 13 C NMR spectra are given in the appendix to Tables 1 and 2. Example 2:
Préparation du O-butanoyl 3 - O - isopropylidene-1,2 α- D-glucofurannose (II)Preparation of O-butanoyl 3 - O - isopropylidene-1,2 α- D-glucofuranose (II)
Dans cet exemple, on prépare le dérivé mono-isopropylidène à partir du dérivé di-isopropylidène (composé I)préparé dans les conditions de l'exemple 1.In this example, the mono-isopropylidene derivative is prepared from the di-isopropylidene derivative (compound I) prepared under the conditions of Example 1.
On place 33 g(0,1 mole) de 0-butanoyl - 3 di - O isopropylidene 1,2 : 5,6αD glucofurannose (butyrate - 3 de diacétone glucose) dans une enceinte thermostatée à 70ºC contenant 500 ml d'H2SO4 0,1 N dans un mélange isopropanol/eau (95/5 v/v).33 g (0.1 mole) of 0-butanoyl - 3 di - O isopropylidene 1.2: 5.6αD glucofuranose (butyrate - 3 of diacetone glucose) are placed in a chamber thermostatically controlled at 70ºC containing 500 ml of H 2 SO 4 0.1 N in an isopropanol / water mixture (95/5 v / v).
Au bout d'une heure, on constate en chromatographie en phase vapeur (C.P.V.) la disparition du pic de butyrate - 3 de di-isopropylidène glucose et la solution est neutralisée par de la tripropylamine dont le sulfate est insoluble dans le milieu. Le solvant est éliminé sous pression réduite. La fraction brute obtenue est purifiée par chromatographie liquide sur silice (Kielselgel 60 Merck 9585, 230 - 400 mesh, 150 g) à l'aide d'un gradient heptane/acétone (fraction 80/20 v/v).After one hour, the vapor phase chromatography (CPV) shows the disappearance of the butyrate-3 peak of di-isopropylidene glucose and the solution is neutralized with tripropylamine, the sulfate of which is insoluble in the medium. The solvent is removed under reduced pressure. The crude fraction obtained is purified by liquid chromatography on silica (Kielselgel 60 Merck 9585, 230 - 400 mesh, 150 g) using a heptane / acetone gradient (80/20 v / v fraction).
Après évaporation sous pression réduite on obtient 20,20 g d'un liquide légèrement jaunâtre qui cristallise F =50 -52ºC et dont le pouvoir rotatoire est de[α]D 22 = + 17,34º (chloroforme).After evaporation under reduced pressure, 20.20 g of a slightly yellowish liquid are obtained which crystallizes F = 50 -52ºC and whose rotary power is [α] D 22 = + 17.34º (chloroform).
Le passage à un gradient eau/THF permet d'isoler une petite quantité de O-butanoyl-3 D-glucopyrannose.The transition to a water / THF gradient makes it possible to isolate a small amount of O-butanoyl-3 D-glucopyranose.
Cette quantité peut être augmentée lorsque l'on opère l'hydrolyse acide ci-dessus à la température plus élevée de 80ºC et en augmentant le temps de contact (3 heures).This quantity can be increased when the above acid hydrolysis is carried out at the higher temperature of 80ºC and by increasing the contact time (3 hours).
Le dérivé] parfaitement caractérisé en spectroscopie RMN (Tableaux 1 et 2) est soluble dans l'eau à raison de plus de 100g/l d'eau à température ambiante. Exemple 3 : Préparation du O-butanoyl-6¬O-isopropylidène-1,2α-D-glucofurannose (III).The derivative] perfectly characterized in NMR spectroscopy (Tables 1 and 2) is soluble in water at a rate of more than 100 g / l of water at room temperature. Example 3: Preparation of O-butanoyl-6¬O-isopropylidene-1,2α-D-glucofuranose (III).
Ce composé est préparé par isomérisation du O-butanoyl-3 O-isopropylidène-1,2α-D-glucofurannose (II) synthétisé dans les conditions de l'exemple 2. On dissout. 30 g de II dans 1 litre de méthanol contenant 10 ml de triéthylamine ; le mélange est agité à température ambiante pendant 2 heures. Après évaporation sous pression réduite, le sirop obtenu, fractionné par HPLC préparative sur colonne WATERS Prep. pack P/N 500-41,75 mesh (éluant hexane-acétone, 80/20 v/v), donne :This compound is prepared by isomerization of O-butanoyl-3 O-isopropylidene-1,2α-D-glucofuranose (II) synthesized under the conditions of Example 2. It is dissolved. 30 g of II in 1 liter of methanol containing 10 ml of triethylamine; the mixture is stirred at room temperature for 2 hours. After evaporation under reduced pressure, the syrup obtained, fractionated by preparative HPLC on a WATERS Prep column. pack P / N 500-41.75 mesh (eluent hexane-acetone, 80/20 v / v), gives:
22 g de produit III, F= 85-86ºC,[α]D 20= -29º (chloroforme)22 g of product III, F = 85-86ºC, [α] D 20 = -29º (chloroform)
5 g de mixte (II + III) 2 g de produit de départ (II), F = 50-52º,[α]D 20 = 17,3º (chloroforme)5 g mixed (II + III) 2 g starting material (II), F = 50-52º, [α] D 20 = 17.3º (chloroform)
La structure du dérivé III est confirmée par RMN (Tableaux 1 et 2).The structure of derivative III is confirmed by NMR (Tables 1 and 2).
Exemple 4 : Préparation du tri-O-butanoyl 3,5,6-0 isopropylidene-1,2α-D-glucofurannose (IV). Ce composé est préparé à partir duExample 4: Preparation of tri-O-butanoyl 3,5,6-0 isopropylidene-1,2α-D-glucofuranose (IV). This compound is prepared from
O-isopropylidène-1,2 α-D-glucofurannose selon la méthode de l'exemple 1 avec un rendement de 62 % après purification sur gel de silice.O-isopropylidene-1,2 α-D-glucofuranose according to the method of Example 1 with a yield of 62% after purification on silica gel.
F = 35-36ºC,[α]D 22 = 10,ºº (chloroforme).F = 35-36ºC, [α] D 22 = 10, ºº (chloroform).
La structure du dérivé IV est confirmée par RMN (Tableaux 1 et 2).The structure of derivative IV is confirmed by NMR (Tables 1 and 2).
Exemple 5 : Préparation du O-n-butanoyl-6 di-O-isopropylidène-1,2: 3,4α-D-galactopyrannose (V).Example 5: Preparation of O-n-butanoyl-6 di-O-isopropylidene-1,2: 3,4α-D-galactopyrannose (V).
Ce composé est préparé à partir du di-O-isopropylidène-1,2 : 3,4α-D galactopyrannose selon la méthode de l'exemple 1 avec un rendement de 62 % après recristallisation dans l'hexane. F = 77-78ºC;[α]D 22 = 40º (chloroforme).This compound is prepared from di-O-isopropylidene-1,2: 3,4α-D galactopyrannose according to the method of Example 1 with a yield of 62% after recrystallization from hexane. F = 77-78ºC; [α] D 22 = 40º (chloroform).
La structure du dérivé 5 est confirmée par RMN (Tableaux 1 et 2). Exemple 6: Préparation du penta O-n-butanoyl-1,2,3,4,6 D-galactopyrannose (VI).The structure of derivative 5 is confirmed by NMR (Tables 1 and 2). Example 6: Preparation of penta O-n-butanoyl-1,2,3,4,6 D-galactopyrannose (VI).
Ce composé est préparé à partir du D-galactose selon la méthode de l'exemple 1, avec un rendement de 83 % après . purification sur gel de silice. nD 20 = 1,488 ;[α]D 22 = + 37º (chloroforme).This compound is prepared from D-galactose according to the method of Example 1, with a yield of 83% afterwards. purification on silica gel. n D 20 = 1.488; [α] D 22 = + 37º (chloroform).
La structure du dérivé VI est confirmée par RMN qui indique en outre la prédominance de l'anomère α (90%) (Tableaux 1 et 2).The structure of derivative VI is confirmed by NMR which also indicates the predominance of the α anomer (90%) (Tables 1 and 2).
Exemple 7: Préparation du penta-O-n butanoyl-1,2,3,4,6 D-glucopyrannose (VII).Example 7: Preparation of penta-O-n butanoyl-1,2,3,4,6 D-glucopyranose (VII).
Ce composé est préparé à partir du D-glucose selon la méthode de l'exemple 1 avec un rendement de 83 % après purification sur gel de silice. nD 20 = 1,4516 ;[α]D 22 = +18,3º (chloroforme). La structure du dérivé VII est confirmée par RMN qui indique en outre la prédominance de l'anomèreα(90%) (Tableaux 1 et 2).This compound is prepared from D-glucose according to the method of Example 1 with a yield of 83% after purification on silica gel. n D 20 = 1.4516; [α] D 22 = + 18.3º (chloroform). The structure of derivative VII is confirmed by NMR which also indicates the predominance of the α anomer (90%) (Tables 1 and 2).
Exemple 8: Préparation du O-n-butanoyl-1-isopropylidène-2,3 glycéryle (VIII). Ce composé est préparé à partir duExample 8: Preparation of O-n-butanoyl-1-isopropylidene-2,3 glyceryl (VIII). This compound is prepared from
O-isopropylidène-1,2 propanol-3 selon la méthode de l'exemple 1, avec .un rendement de 51 % après purification sur gel de silice. liquide, nD 20 = 1,4295,O-isopropylidene-1,2 propanol-3 according to the method of Example 1, with a yield of 51% after purification on silica gel. liquid, n D 20 = 1.4295,
La structure du dérivé VIII est confirmée par RMN (Tableaux 1 et 2).The structure of derivative VIII is confirmed by NMR (Tables 1 and 2).
Exemple 9 : Préparation du O-n butanoyl-1 propane diol-2,3 (IX).Example 9: Preparation of O-n butanoyl-1 propane diol-2,3 (IX).
Ce composé est préparé à partir de VIII par déprotection des hydroxyles en 2 et 3 soit en phase homogène selon la méthode de l'exemple 2 légèrement modifiée (6 heures à température ambiante au lieu de 1 heure à 70ºC) ; soit en phase hétérogène par passage sur une colonne de 25 ml de résine Amberlist. 15 Wet maintenue à 30ºC avec un débit de 0,1 ml/mn de VIII en solution à 20 % dans un mélange isopropanol-eau 90-10 v/v. La purification s'effectue sur colonne de gel de silice avec comme éluant l'éther éthylique.This compound is prepared from VIII by deprotection of the hydroxyls in 2 and 3, ie in the homogeneous phase according to the method of Example 2 slightly modified (6 hours at room temperature instead of 1 hour at 70 ° C); either in the heterogeneous phase by passage through a column of 25 ml of Amberlist resin. 15 Wet maintained at 30ºC with a flow rate of 0.1 ml / min of VIII in 20% solution in an isopropanol-water mixture 90-10 v / v. The purification is carried out on a column of silica gel with ethyl ether as eluent.
Les rendements en produit final sont de 53 % en phase homogène et. de 85 % en phase hétérogène, après purification sur gel de silice. liquide, nD 20 = 1,4465.The yields of final product are 53% in the homogeneous phase and. 85% in heterogeneous phase, after purification on silica gel. liquid, n D 20 = 1.4465.
La structure du dérivé IX est confirmée par RMN (Tableaux 1 et 2).The structure of the derivative IX is confirmed by NMR (Tables 1 and 2).
Exemple 10. Préparation du O-butanoyl-3 di-O-palmitoyl-1,2 glycéryle (X).Example 10. Preparation of O-butanoyl-3 di-O-palmitoyl-1,2 glyceryl (X).
Ce composé est préparé à partir du composé IX selon la méthode de l'exemple 1 avec un rendement de 72 % après purification sur gel de silice.This compound is prepared from compound IX according to the method of Example 1 with a yield of 72% after purification on silica gel.
F = 54ºC. La structure du dérivé X est confirmée par RMNF = 54ºC. The structure of the derivative X is confirmed by NMR
(Tableaux 1 et 2).(Tables 1 and 2).
Exemple 11 : Préparation du di-O-butanoyl-2,3 O-palmitoyl-1-glycéryle (XI).Example 11: Preparation of di-O-butanoyl-2,3 O-palmitoyl-1-glyceryl (XI).
Ce composé est préparé selon la séquence suivante : a) estérification par le chlorure de palmitoyle duThis compound is prepared according to the following sequence: a) esterification with palmitoyl chloride of
O-isopropylidène-1,2 propanol-3, selon la méthode de O-isopropylidene-1,2 propanol-3, according to the method of
l'exemple 1 conduisant au O-palmitoyl-1 O-isopropylidène-2,3 propane ; b) déprotection des hydroxyles en 2 et 3 du composé obtenu en a) selon les méthodes décrites pour la préparation de IX, conduisant au O-palmitoyl-1 propane diol-2,3 ; c) estérification par le chlorure de butanoyle du composé obtenu en b) selon la méthode de l'exemple 1 conduisant à XI, avec un rendement de 70 % après purification sur gel de silice. Liquide, nD 20 = 1,4475.Example 1 leading to O-palmitoyl-1 O-isopropylidene-2,3 propane; b) deprotection of the hydroxyls in 2 and 3 of the compound obtained in a) according to the methods described for the preparation of IX, leading to O-palmitoyl-1 propane diol-2,3; c) esterification with butanoyl chloride of the compound obtained in b) according to the method of Example 1 leading to XI, with a yield of 70% after purification on silica gel. Liquid, n D 20 = 1.4475.
La structure du dérivé XI est confirmée par RMN (Tableaux 1 et 2)The structure of derivative XI is confirmed by NMR (Tables 1 and 2)
Tests pharmacologigues et pharmacocinétiques. 1º TOXICITE - les toxicités aiguës chez la souris SWISS et chroniques chez le rat albinos sont déterminées selon les protocoles normalisés habituels :Pharmacological and pharmacokinetic tests. 1 ° TOXICITY - acute toxicities in SWISS mice and chronic toxicities in albino rats are determined according to the usual standardized protocols:
COMPOSES TOXICITETOXICITY COMPOUNDS
DL50 en g/kg voie i.p. voie oraleLD50 in g / kg i.p. oral
I 1,75 ± 0,39 > 8I 1.75 ± 0.39> 8
II 4,2 ± 0,4 > 8II 4.2 ± 0.4> 8
III 3,00 ± 0,35 > 8III 3.00 ± 0.35> 8
IV > 8 > 8IV> 8> 8
V 2,5 ± 0,4 > 8V 2.5 ± 0.4> 8
VI > 8 > 8VI> 8> 8
VII > 8 > 8VII> 8> 8
VIII 5,4 ± 0,3 > 8VIII 5.4 ± 0.3> 8
IX 2,25 ± 0,23 > 8IX 2.25 ± 0.23> 8
X > 2 > 6X> 2> 6
XI > 8 > 8XI> 8> 8
L'administration de 1,12 g/kg de Composé I chez le rat ne provoque aucune létalité. 2º tests pharmacocinétiques chez le lapin.The administration of 1.12 g / kg of Compound I in rats does not cause any lethality. 2nd pharmacokinetic tests in rabbits.
Ces tests sont réalisés de la manière suivante : On prépare une solution contenant 0,600 g du composé selon l'invention dans 2 ml de glycérol formai (mélange de dioxanne - 1,3 ol-5 et de dioxolanne - 1,3 méthanol-4). On procède sur les lapins d'expérience, à une injection préalable (0,1 ml/kg) d'une solution d'héparine à la concentration de 5000 U.I./ml dans la veine marginale de l'oreille ; l'aiguille est laissée en place permettant d'administrer, deux minutes plus tard, une dose de 0,1 ml/kg (soit 23 mg/kg) de la solution préparée ci-dessus. Des prélèvement sanguins de 1 cm3 échelonnés dans le temps sont effectués au niveau de la veine marginale de l'autre oreille. Le sang est centrifugé (2 000 tours/min durant 3 min) et le sérum est recueilli.These tests are carried out as follows: A solution containing 0.600 g of the compound according to the invention is prepared in 2 ml of formal glycerol (mixture of dioxane - 1.3 ol-5 and dioxolane - 1.3 methanol-4). On experimental rabbits, a prior injection (0.1 ml / kg) of a heparin solution at a concentration of 5000 IU / ml is carried out in the marginal vein of the ear; the needle is left in place allowing administration, two minutes later, of a dose of 0.1 ml / kg (i.e. 23 mg / kg) of the solution prepared above. Blood samples of 1 cm3 spread over time are taken from the marginal vein of the other ear. The blood is centrifuged (2,000 rpm for 3 min) and the serum is collected.
La concentration en acide butyrique potentiel sérique est déterminée par chromatographie en phase gazeuse à l'aide d'une colonne de CHROMOSORB 101 ou de CARBOPACK C.The concentration of potential serum butyric acid is determined by gas chromatography using a column of CHROMOSORB 101 or CARBOPACK C.
L'échantillon à analyser par chromatographie est préparé par le traitement suivant : - 80 ul de sérum - 10 ul d'acide formique purThe sample to be analyzed by chromatography is prepared by the following treatment: - 80 μl of serum - 10 μl of pure formic acid
10 ul d'acide valérique (étalon interne) à la concentration de 50 mg/1.10 μl of valeric acid (internal standard) at a concentration of 50 mg / 1.
Ce traitement avant la chromatographie, a pour effet d'hydrolyser le composé ou ses métabolites et de libérer l'acide n-butyrique.This treatment before chromatography has the effect of hydrolyzing the compound or its metabolites and releasing n-butyric acid.
La quantification se fait par comparaison des surfaces respectives de l'acide butyrique et de l'acide valérique, les coefficients de réponse ayant été déterminés préalablement. On mesure ainsi la concentration sérique (en mmole) d'acide n-butyrique potentiel ou libérable à partir du composé ou ses métabolites contenus dans le sérum. Ces mesures permettent de déterminer la demi-vie sérique apparente du composé ester injecté.The quantification is done by comparison of the respective surfaces of butyric acid and valeric acid, the response coefficients having been determined beforehand. The serum concentration (in mmoles) of potential or releasable n-butyric acid is thus measured from the compound or its metabolites contained in the serum. These measurements make it possible to determine the apparent serum half-life of the injected ester compound.
Ces mesures donnent les résultats suivants consignés dans le tableau 3.These measurements give the following results, recorded in Table 3.
Les composés selon l'invention ont une durée de The compounds according to the invention have a duration of
demi-vie, dans le courant sanguin du lapin, beaucoup plus longue que celle des butyrates salins. Il constituent ainsi une réserve potentielle en acide butyrique, par hydrolyse enzymatique lente à l'aide des nombreuses estérases présentes chez l'animal et l'homme.half-life in rabbit bloodstream much longer than that of saline butyrates. They thus constitute a potential reserve of butyric acid, by slow enzymatic hydrolysis using the many esterases present in animals and humans.
3º Tests antitumoraux in-vivo chez la souris. Les études des propriétés antitumorales ont été menées chez la souris, selon le protocole de CHANY et CERUTTI (Int. J. Cancer, 1982, 30, 489) sur les composés I à V ci-dessus. Pour obtenir des souris avec tumeur (s) on utilise la souche TG 180 (injection par voie i.p. de 106 cellules pour une souris mâle de 25 g et de 0,1 ml d'une solution de Corynebacterium Parvum Mérieux (CP)). Première expérience Cette expérience porte sur 135 souris SWISS divisée en3º In-vivo anti-tumor tests in mice. The anti-tumor properties studies were carried out in mice, according to the protocol of CHANY and CERUTTI (Int. J. Cancer, 1982, 30, 489) on the compounds I to V above. To obtain mice with tumor (s), the TG 180 strain is used (ip injection of 10 6 cells for a 25 g male mouse and 0.1 ml of a solution of Corynebacterium Parvum Mérieux (CP)). First experiment This experiment concerns 135 SWISS mice divided into
9 lots de 15 souris chacun. Après injection au jour JO de cellules TG 180 et de CP, on procède de la manière suivante: Le lot témoin nº 1 est. traité quotidiennement par une injection d'un placebo. Le lot témoin nº 2 est traité quotidiennement par une injection de CP (0,1 ml).9 lots of 15 mice each. After injection on day D O of TG 180 cells and of CP, the procedure is as follows: The control batch No. 1 is. treated daily with a placebo injection. Control batch 2 is treated daily with an injection of CP (0.1 ml).
Le lot témoin nº 3 " " " par une injection d'interféron EAT-113, 0,5 ml.Control lot 3 "" "by an injection of interferon EAT-113, 0.5 ml.
Le lot témoin nº 4 est traité chaque jour alternativement par du CP puis de l'interféron aux-mêmes doses que ci-dessus.Control batch 4 is treated daily with CP and then interferon at the same doses as above.
Les lots expérimentaux sont traités, alternativement à J + 1 par le composé selon l'invention (0,5 ml d'une solution 8 mmolaire en composé I à V), à J+2 par de l'interféron, à J+3 par du CP et ceci pendant trois semaines.The experimental batches are treated, alternately on D + 1 with the compound according to the invention (0.5 ml of an 8 mmolar solution in compound I to V), on D + 2 with interferon, on D + 3 by CP and this for three weeks.
On note le nombre de survivants et la présence ou non de tumeur (tumeur + ou tumeur -) chez ces survivants cinq semaines après JO. Le tableau 4 ci-après donne les résultats observés : Le tableau 4 montre la très bonne efficacité thérapeutique antitumorale des composés I, II, III selon l'invention, par rapport aux lots témoins nº 1, 2, 3 et 4 . La quantité d'acide n-butyrique potentiel pouvant se libérer dans l'organisme de la souris est dans ce protocole pour les composés I, II, III de 14 mg par kilogramme de souris, ce qui correspond en théorie à la dose optimale préconisée pour le butyrate d'arginine pour une souris de 25 g. Deuxième expérience.Note the number of survivors and the presence or absence of tumor (tumor tumor + or -) in these survivors five weeks after J O. Table 4 below gives the results observed: Table 4 shows the very good therapeutic anti-tumor efficacy of the compounds I, II, III according to the invention, compared with the control batches 1, 2, 3 and 4. The amount of potential n-butyric acid which can be released in the mouse body is in this protocol for compounds I, II, III of 14 mg per kilogram of mouse, which corresponds in theory to the optimal dose recommended for arginine butyrate for a 25 g mouse. Second experience.
Des essais selon le même protocole que ci-dessus pour déterminer l'effet-dose des composés selon l'invention ont été effectués en maintenant constante la quantité de CP et d'interféron de l'expérience précédente, mais en faisant varier la concentration de 0,25 à 50 millimoles en composé II par litre de solution injectable (0,5 ml de cette solution étant injectée) soit l'équivalent potentiel de 0,44 à 88 mg d'acide n-butyrique par kg de souris.Tests according to the same protocol as above to determine the dose-effect of the compounds according to the invention were carried out by keeping constant the amount of CP and of interferon from the previous experiment, but by varying the concentration of 0.25 to 50 millimoles of compound II per liter of solution for injection (0.5 ml of this solution being injected), ie the potential equivalent of 0.44 to 88 mg of n-butyric acid per kg of mouse.
La survie à 100 jours est de 1 souris sur 45 pour le CP seul, 0 sur 45 pour l' interféron seul, 30 sur 45 pour le composé II, quel que soit sa concentration entre 0,25 et 50 mmol/l, le composé II étant administré en alternance avec CP et interféron selon le protocole ci-dessus.The survival at 100 days is 1 mouse out of 45 for CP alone, 0 out of 45 for interferon alone, 30 out of 45 for compound II, whatever its concentration between 0.25 and 50 mmol / l, the compound II being administered alternately with CP and interferon according to the above protocol.
Un autre essai, selon un autre protocole où 1' interféron a été supprimé mais où l'on continue à administrer en alternance 0,1 ml de CP et 0,5 ml d'une solution de composé II à une concentration variant de 0,25 à 50 millimoles/l, montre que le nombre de souris survivantes à 100 jours est constant quelle que soit la concentration en composé II et s'élève en moyenne à 20 sur 45 souris.Another trial, according to another protocol in which the interferon has been suppressed but in which one continues to administer alternately 0.1 ml of CP and 0.5 ml of a solution of compound II at a concentration varying from 0, 25 to 50 millimoles / l, shows that the number of mice surviving to 100 days is constant whatever the concentration of compound II and amounts on average to 20 out of 45 mice.
Ces résultats montrent qu'une dose de composé II au plus égale à l'équivalent de 0,44 mg d'acide n-butyrique potentiel par kilogramme de souris est suffisant pour obtenir l'effet thérapeutique maximum. Cette dose est environ 32 fois inférieure à la dose optimale préconisée de butyrate d'arginine (14 mg en acide butyrique) et par conséquent permet d'éviter des doses These results show that a dose of compound II at most equal to the equivalent of 0.44 mg of potential n-butyric acid per kilogram of mouse is sufficient to obtain the maximum therapeutic effect. This dose is approximately 32 times lower than the optimal recommended dose of arginine butyrate (14 mg in butyric acid) and therefore makes it possible to avoid doses
thérapeutiques massives chez l'homme. Rappelons que la nécessité actuelle de dose massive de butyrate d'arginine entraîne des effets secondaires chez l'homme, du genre hyperammoniémie avec baisse de la sécrétion d'insuline et perturbations du cycle de l'urée et du métabolisme du glucose (c.f Brazier et coll, Chimica Clinica Acta page 261, vol 148, 1988).massive therapeutic in humans. Recall that the current need for a massive dose of arginine butyrate causes side effects in humans, such as hyperammonemia with reduced insulin secretion and disturbances in the urea cycle and glucose metabolism (see Brazier and coll, Chimica Clinica Acta page 261, vol 148, 1988).
Le fait également que l'association composé II plus CP seul donne d'aussi bons résultats thérapeutiques que l'association interferon plus CP plus butyrate d'arginine permet d'envisager grâce aux composés selon l'invention une suppression de l'interferon, alors que l'association du CP plus le butyrate d'arginine sans l'interferon, a une efficacité thérapeutique faible (8 souris survivantes sur 45 à 100 jours).The fact also that the combination of compound II plus CP alone gives as good therapeutic results as the combination of interferon plus CP plus arginine butyrate makes it possible to envisage, thanks to the compounds according to the invention, a suppression of interferon, then that the association of CP plus arginine butyrate without interferon, has a low therapeutic efficacy (8 surviving mice over 45 to 100 days).
D'autres effets thérapeutiques ont été mis en évidence pour les composés selon l'invention en particulier le composé II c'est ainsi que : a) Le composé II montre in vivo une potentialisation de l'action de l'interféron EAT-113 sur le virus Herpès hominis type II ; b) l'association du composé II et de l' interferon EAT-113 a une action in vitro plus forte que l'association butyrate d'arginine et interferon EAT-113 sur les cellules mammaires néoplasiques (souche NCF 410) ; c) le composé II à une action plus forte in vitro sur les cellules infectées par le virus du sida murin TR-26 que le butyrate d'arginine ; d) les composés selon l'invention peuvent être associés à d'autres produits, notamment le methotrexate, en vue du traitement thérapeutique des tumeurs.Other therapeutic effects have been demonstrated for the compounds according to the invention, in particular compound II. This is how: a) Compound II shows in vivo a potentiation of the action of the interferon EAT-113 on Herpes hominis type II virus; b) the combination of compound II and the interferon EAT-113 has a stronger in vitro action than the combination of arginine butyrate and interferon EAT-113 on neoplastic breast cells (strain NCF 410); c) compound II has a stronger action in vitro on cells infected with the murine AIDS virus TR-26 than arginine butyrate; d) the compounds according to the invention can be combined with other products, in particular methotrexate, for the therapeutic treatment of tumors.
En fonction des résultats pharmacologiques exposés ci-dessus, des compositions sont préparées en vue d'une utilisation thérapeutique, chez l'homme ou l'animal comprenant 100 à 2000 mg d'un composé selon l'invention et tout excipient pharmaceutiquement acceptable. Ainsi par exemple le composé II est mis en solution isotonique avec du sérum physiologique ou glucose pour former un médicament injectable dosé unitairement entre 100 à 500 mg de composé II. Le composé II est mélangé éventuellement à un excipient tel qu'un polyéthylène glycol(PEG) pour former un médicament administrable par voie orale ou rectale et dosé unitairement entre 1 et 2 g de composé II. Pour l'administration orale il est prévu des gélules gastrorésistantes pour éviter l'hydrolyse acide dans l'estomac. Depending on the pharmacological results set out above, compositions are prepared for therapeutic use in humans or animals comprising 100 to 2000 mg of a compound according to the invention and any pharmaceutically acceptable excipient. Thus, for example, compound II is placed in isotonic solution with physiological serum or glucose to form an injectable drug dosed unitarily between 100 to 500 mg of compound II. Compound II is optionally mixed with an excipient such as a polyethylene glycol (PEG) to form a medicament which can be administered orally or rectally and dosed individually between 1 and 2 g of compound II. For oral administration, gastro-resistant capsules are provided to avoid acid hydrolysis in the stomach.

Claims

REVENDICATIONS 1. Composé de formule :CLAIMS 1. Compound of formula:
W-(O-CO-CH2-CH2-CH3)n dans laquelle W représente le radical d'un produit ayant au moins un groupe OH estérifiable par l'acide n-butyrique, n pouvant varier de 1 au nombre total des groupes OH estérifiables dans ledit produit.W- (O-CO-CH 2 -CH 2 -CH 3 ) n in which W represents the radical of a product having at least one OH group esterifiable by n-butyric acid, n possibly varying from 1 to the total number esterifiable OH groups in said product.
2. Composé selon la revendication 1, caractérisé en ce que ledit produit est choisi parmi les carbohydrates et leurs dérivés acétonides, les carbohydrates hydrogénés et leurs dérivés acétonides, les esters glycéryliques d'acides gras, le dérivé acétonide du glycérol, la sérine et l'homosérine.2. Compound according to claim 1, characterized in that said product is chosen from carbohydrates and their acetonide derivatives, hydrogenated carbohydrates and their acetonide derivatives, glyceryl esters of fatty acids, acetonide derivative of glycerol, serine and l 'homoserine.
3. Composé selon la revendication 1 ou 2, caractérisé en ce que le carbohydrate est un monosaccharide aldose ou cétose, cyclique ou acyclique.3. Compound according to claim 1 or 2, characterized in that the carbohydrate is an aldose or ketose, cyclic or acyclic monosaccharide.
4. L'O-butanoyl-3-di-O-isopropylidène 1,2 : 5,6 α-D-glucofurannose.4. O-butanoyl-3-di-O-isopropylidene 1,2: 5,6 α-D-glucofuranose.
5. L'O-butanoyl-3-O-isopropylidène-1,2 α-D-glucofurannose. 5. O-butanoyl-3-O-isopropylidene-1,2 α-D-glucofuranose.
6. L'O-butanoyl-3-D-glucopyrannose.6. O-butanoyl-3-D-glucopyranose.
7. L'O-butanoyl-6-O-isopropylidène-1,2 α-D-glucofurannose.7. O-butanoyl-6-O-isopropylidene-1,2 α-D-glucofuranose.
8. Le tri-O-butanoyl-3,5,6 O-isopropylidène-1 , 2 α-D-glucofurannose. 8. Tri-O-butanoyl-3,5,6 O-isopropylidene-1,2,2 α-D-glucofuranose.
9. Le O-butanoyl-6 di-O-isopropylidène 1,2 : 3,4 α-D-galactopyrannose.9. O-butanoyl-6 di-O-isopropylidene 1,2: 3,4 α-D-galactopyrannose.
10. Le penta O-butanoyl-1,2,3,4,6 D-galactopyrannose.10. The penta O-butanoyl-1,2,3,4,6 D-galactopyrannose.
11. Le penta-O-butanoyl-1,2,3,4,6 D-glucopyrannose.11. Penta-O-butanoyl-1,2,3,4,6 D-glucopyranose.
12. Le O-butanoyl-1-isopropylidène-2,3 glycéryle. 12. O-butanoyl-1-isopropylidene-2,3 glyceryl.
13. Le O-butanoyl-3-di-0-palmitoyl-1,2 glycéryle.13. O-butanoyl-3-di-0-palmitoyl-1,2 glyceryl.
14. Le di-n butanoyl-2,3 O-palmitoyl-1-glycéryle.14. Di-n butanoyl-2,3 O-palmitoyl-1-glyceryl.
15. Composé selon l'une quelconque des revendications précédentes, caractérisé en ce que sa demi-vie sérique chez le lapin est comprise entre 1,75 heure et 24 heures. 15. Compound according to any one of the preceding claims, characterized in that its serum half-life in rabbits is between 1.75 hours and 24 hours.
16. Composé selon l'une quelconque des revendications précédentes pour une. utilisation comme médicament.16. Compound according to any one of claims previous for one. use as medicine.
17. Composé selon l'une quelconque des revendications 1 à 15, associé a au moins une substance choisie parmi le corynebactérium parvum, l'interféron (EAT 113), le methotrexate, pour une utilisation comme médicament.17. Compound according to any one of claims 1 to 15, associated with at least one substance chosen from corynebacterium parvum, interferon (EAT 113), methotrexate, for use as a medicament.
18. Utilisation d'un composé selon l'une quelconque des revendications 1 à 15 avec tout excipient pharmaceutiquement acceptable pour l'obtention d'un médicament destiné à une utilisation antitumorale et/ou antivirale. 18. Use of a compound according to any one of claims 1 to 15 with any pharmaceutically acceptable excipient for obtaining a medicament intended for antitumor and / or antiviral use.
EP88908255A 1987-09-25 1988-09-26 Esters of n-butyric acid and their use as drugs Withdrawn EP0352301A1 (en)

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FR8713294 1987-09-25
FR8713294A FR2621039B1 (en) 1987-09-25 1987-09-25 NON-TOXIC N-BUTYRIC ACID DERIVATIVES WITH DELAYED THERAPEUTIC ACTIONS
FR8809092 1988-07-05
FR8809092A FR2633929B2 (en) 1987-09-25 1988-07-05 NON-TOXIC N-BUTYRIC ACID DERIVATIVES WITH DELAYED THERAPEUTIC ACTIONS

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