WO2005077963A1

WO2005077963A1 - Saccharide and itol derivatives having an o-alkyl group or an o-alkyl group and an o-n butanyl group, uses as medicines in tumoral or benign proliferative pathologies

Info

Publication number: WO2005077963A1
Application number: PCT/FR2004/000079
Authority: WO
Inventors: Gerard Andre Daniel Goethals; Vincent Yves Olivier Jules Lequart; Patrick Emile Marius Martin; Jean Claude Maziere; Cecile Maziere; Philippe Rene Michel Puillart; Pierre Joseph Villa
Original assignee: Institut Superieur Agricole De Beauvais
Priority date: 2004-01-16
Filing date: 2004-01-16
Publication date: 2005-08-25

Abstract

The invention concerns saccharide and itol derivatives having an O-alkyl group or an O-alkyl group and an O-n butanyl group, uses as medicines in tumoral or benign proliferative pathologies. One of the aims of the present invention is to provide for the synthesis of saccharide and itol derivatives having a O-alkyl group of general formula (I) or having an O-alkyl group and an O-n butanyl group of general formula (II) wherein: Su represents a saccharide derivative; R represents a C8-C18 n-alkyl or n-alkenyl chain; P represents a group of atoms bound to the oxygen atom of the hydroxy group; i represents the number of hydroxylated sites not bearing R as defined above, nor a butanyl group, said hydroxylated groups capable of being all (j=0) or partly free (0<j<i) or still protected in the form of O-P groups; j represents the number of protected hydroxylated groups in the form of O-P groups. In another embodiment, the invention concerns the use of said derivatives as compounds for preparing food or drug additives, useful in particular in the treatment of cancer and in the treatment of tumoral or benign proliferative pathologies or still in the preparation of potentiating agents enhancing the efficacy of antitumor photochemotherapy.

Description

Derivatives of saccharides and itols having an O-alkyl group or an O-alkyl group and an O-n-butanoyl group. Applications as medicaments in proliferative tumors or benign pathologies.

The present invention relates to saccharide and itol derivatives having an O-alkyl group or an O-alkyl group and an On-butanoyl group and their applications as medicaments in proliferative tumoral or benign pathologies. It is known that butyric acid and its salts have an antiproliferative action on certain types of tumors (Prasad, KN & al., 1980, Life Sciences, 27, 1351; Fisckhoff & al., Leukemia, 1990, 4, 302; Samid, D. & al., Cancer Res., 1992, 52, 1988). However, the rapid elimination of these substances by the body, as well as the need to maintain high plasma concentrations (Daniel, P. & al., Clin.Chim.Acta, 1989, 81, 255), impose serious constraints. which considerably limit the use of these drugs in oncology. However, these products having an undeniable potential interest in anticancer therapy, various strategies have been developed with the aim of modifying their pharmacokinetics, and in particular of lengthening their plasma lifespan. Among these strategies, it has been proposed to prepare derivatives of butyric esters of oses (glucose, mannose) and of glycerol (Pieri & al., Patent FR 871 294, 1987; patent FR 8809092, 1988). With this in mind, the butyric ester was conceived as the precursor of the active principle (butyric acid), de-esterification taking place in various compartments, in particular at the intracellular level, most of the cells having very active cytosolic esterases. It is also known that the cell membrane, due to its hydrophobic nature, is not very permeable to charged molecules, such as organic acids or proteins (Singer, SJ, Nicolson, GL 1972, Science, 175. 720; Bretscher, M., 1985, Sci. Am., 253, 100; Petty, HR, 1993, in Molecular Biology of Membranes: Structure and Functions, Plénum Press, NY; Yeagle, PL 1993, in The Membranes of Cells, Académie Press, San Diego) and that the presence of a lipophilic chain facilitates the transmembrane passage of charged substances, even bulky ones, such as proteins (Kabanov, AN. & al. 1989, Prot. Eng., 3, 39). Finally, it is known that certain derivatives of alkyl chains can act on the activity of membrane ion channels (Becaert, T. & al. 1996, Boll. Chim. Farm. 135. 204) some of which play a role in controlling the cell proliferation (Naur, S. & al., 1998, J. Cell Physiol., 177. 402; Wiecha, J. & al. 1998, J. Nasc. Res., 35, 363). One of the aims of the present invention is to provide the synthesis of new saccharide and itol derivatives having an O-alkyl group and corresponding to the general formula I and or also having an O-alkyl group and an O- n- group. butanoyl and corresponding to general formula II, the latter representing a subset of the type I compounds:

II in which Su represents a cyclic or unprotected saccharide derivative, protected or not, chosen for example from hexose, pentose, deoxyhexose, or else an itol derivative, cyclic or not, protected or not corresponding for example to glycerol, in which R represents an n-alkyl or n-alkenyl chain having 8 to 18 carbon atoms, in which P represents a group of atoms linked to the oxygen atom of the hydroxyl group and forming, via the latter, and with Su, an ether function, P being preferably chosen from benzyl or trityl groups, or alternatively forming an ester function, P being preferably chosen from acetyl and benzoyl groups, or alternatively forming an acetal of dioxolane type, preferably isopropylidene or benzylidene, in which i represents the number of hydroxylated sites not carrying R as defined above, these hydroxylated groups possibly being all Q = 0) or partly free (0 <j <i) or even prot aged in the form of OP groups with P as defined above, in which j represents the number of protected hydroxyl groups in the form of OP groups, the j groups P can be identical or not. The type II family represents a subset of type l compounds. The type I compounds in accordance with the invention, other than the alkyl glycosides, can be prepared according to the method of the literature (F. CHELLE, G. RONCO, P. VILLA, Patent FR 87 05277, PCT / FR8800128, WO8808000). The alkyl glycosides of general formula I are prepared with the β-D or α- L configuration, according to scheme 1,

Su (OH), Scheme 1. Synthesis of type I alkyl glycosides in which the carbon C-2 of hexose or deoxyhexose Su (OH) _ι is linked to R '= OH and to R "= H or at R '= H and at R "= OH, and in which ROH represents an n-alkanol having 1 to 18 carbon atoms: - by reacting at room temperature and with stirring an equivalent of hexose or deoxyhexose with five to ten equivalents of n-alkanol in the presence preferably of one to two equivalents of acetyl chloride. After ten to twenty hours of reaction, the mixture is brought to 0 ° C., a volume equivalent to 5 to 10 times the volume of the initially introduced n-alkanol, of a non-polar solvent is added and the following is collected by filtration: pure alkyl glycoside, of configuration β-D or -L. The non-polar solvent may be a linear or branched alkane preferably chosen from pentane, hexane or heptane or alternatively a cyclic or acyclic ether and preferably ethyl ether or a mixture of ethyl ether-hexane. This process has the advantage of being distinguished from the TAKEO method (K. TAKEO, M. NAKAGEN, Y. TERRAMOTO, Carbohydrate Research, 1990, 201. 261) in that ROH is an n-alkanol having 8 to 18 atoms carbon, instead of allyl alcohol and the products are obtained pure with the β-D or α-L configuration, without neutralization with a base (NaHCO ₃ ), neither washing with water, nor extraction with toluene, followed by evaporation and recrystallization from ethanol. The type II compounds can be prepared from the type I compounds, with the exclusion of alkyl glycosides, by regioselective grafting of the On-butanoyl group according to scheme 2,

Scheme 2. Synthesis of type II compounds One equivalent of type I compound is reacted at room temperature with stirring in a non-polar solvent, with one equivalent of n-butanoyl chloride in the presence of 1.1 equivalents of a weak mineral or organic base; the non-polar solvent may be a cyclic, acyclic, branched or unbranched alkane, or else an aromatic hydrocarbon or a mixture of the two or better toluene; the weak base can be either mineral chosen, for example, from alkaline earth carbonates or alkali carbonates and hydrogen carbonates, or organic chosen, for example, from cyclic or acyclic amines and preferably triethylamine. After consumption of the reagents, the solution is filtered and the solvent evaporated under reduced pressure and the type II product obtained, is purified by selective solubilization, or recrystallization or chromatography on neutral silica. When type II products are obtained from hexoses and pentitols, the reaction according to scheme 2 is carried out on type I compounds having the isopropylidene protective group. In order to then obtain the completely deprotected type II compounds (j = 0), an acid-catalyzed deacetalization is carried out in a hydroxylated solvent chosen for example from water or an alkanol preferably chosen from methanol or ethanol, or a water-alkanol mixture, or a mixture of water and non-polar solvent preferably chosen from dioxane or THF. The deacetalization can be carried out: - either in a homogeneous medium in the presence of a mineral acid preferably chosen from H ₂ SO ₄ or HC1 or in the presence of an organic acid preferably chosen from HOTs, CH ₃ COOH or CF ₃ COOH, followed by possible neutralization with a weak base then washing, evaporation of the solvent and purification, - either in a heterogeneous medium in the presence of acid resin chosen, preferably from DOWEX or AMBERLYST H ⁺ , followed by filtration, evaporation of the solvent and purification. The purification can be carried out by selective solubilization, recrystallization or chromatography on neutral silica. Another object of the present invention is the use of saccharide and itol derivatives having an O-alkyl group and corresponding to the general formula I or also having an On-butanoyl group and an O-alkyl group corresponding to the general formula II, the latter representing a subset of the type I compounds:

II with Su, R, P, i and j as defined above and in accordance with the present invention, as compounds for the preparation of food additives or of drugs, usable in particular in oncology and in the treatment of proliferative tumoral or benign pathologies or else the preparation of potentiating agents making it possible to improve the effectiveness of anti-tumor photochemotherapy. The drugs thus prepared may be included in the arterial prostheses. By way of examples and without this being considered as limiting, the following is a description of the synthesis of some derivatives in accordance with the invention as well as a toxicological study and antiproliferative tests on malignant and non-malignant cell lines. Example 1. Preparation of 10-alkyl-D, L-glycerols la-c (type I). 26.4 g (0.20 mol) of l, 2-O-isopropylidene-D, L-glycerol (solketal) are reacted with 0.22 mol of alkyl bromide in the presence of 28 g (0.5 mol ) of powdered KOH in 300 mL of the toluene-DMSO 80:20 v / v mixture at room temperature for 24 hours. After filtration, washing with an aqueous solution of NH ₄ C1 at 20%, the organic phase is evaporated under reduced pressure and the O-alkylated derivatives are isolated in the pure state by chromatography on neutral silica (eluent hexane-acetone 95: 5 v / v). Table 1.

¹³ C NMR; 71.7

6.2 (CMe ₂ ), 71.7 (C-α), 22.5, 31.7 (C-β, C-ω-1), 13.9 (C-ω). The O-alkylated derivatives are then deacetalized in water-ethanol 95 ° GL with 1 g of Amberlyst 15H ⁺ resin per gram of acetal, at 50 ° C for 4 h. After filtration, evaporation of the solvent, the compounds la-c are isolated in the pure state by chromatography on neutral silica (eluent hexane-acetone 80:20 v / v). Table 2.

Example 2. Preparation of l-O-alkyl-2,3-O-isopropylidene-D, L-xylitols 2a-c and l-O-alkyl-D, L-xylitols 3a-c (type I). 23.2 g (0.10 mol) of 2,3: 4,5-di-O-isopropylidene-D, L-xylitol is reacted with

0.11 mol of alkyl bromide in the presence of 14 g (0.25 mol) of powdered KΟH in

300 mL of toluene-DMSO 80:20 v / v mixture at room temperature for 24 hours.

After filtration, washing with an aqueous solution of NH ₄ C1 at 20%, the organic phase is evaporated under reduced pressure and the O-alkylated derivatives are isolated in the pure state by chromatography on neutral silica (eluent hexane-acetone 95: 5 v / v).

Table 3.

The O-alkylated derivatives are then deacetalized in water-ethanol 95 ° GL with 2 g of Amberlyst 15H ⁺ resin per gram of diacetal, at 50 ° C. After disappearance of the diacetal, filtration and evaporation of the solvent, the compounds 2a-c and 3a-c are isolated at pure state by chromatography on neutral silica (hexane-acetone gradient 80:20 v / v to 20:80 v / v).

Example 3. Preparation of alkyl aL-fucopyranosides 4a-b, alkyl β-D-galactopyranosides 5a-b and alkyl β-D-glucopyranosides 6a-b (type I). 0.10 mol of L-fucose is reacted at room temperature with stirring with 9 equivalents of alkanol nC _n H _{2n +} ιOH in the presence of 2 equivalents of CH ₃ COCl with stirring for 20 h. The mixture is then brought to 0 ° C. and 50 ml of ethyl ether are added. The crystallized pure glycosidic derivatives of α-L configuration for fucose are separated by filtration. The β-D-galactopyranoside derivatives of alkyl 5a-b and β-D-glucopyranoside derivatives of alkyl 6a-b were prepared by the Schmidt method using peracetate. The β configuration derivatives of galactose and glucose are obtained by chromatographic purification.

Table 4.

NMR 4b (C ₅ D ₅ N). ¹³ C: 100.9 (C-1); 73.5 (C-4); 71.5 (C-3); 70.4 (C-2); 68.4 (C-α; 67.3 (C-5); 30.3 - 23.1 (CH ₂ ); 17.4 (C-6); 14.4 (CH ₃ ). 5a NMR (DMSO). ¹³ C: 104.3 (C-1; 75.9 (C-5); 74.3 (C-3); 71.4 (C-2); 69.3 (C-α);

69.0 (C-4); 61.2 (C-6); 32.1 - 22.9 (CH ₂ ) _n ; 14.8 (CH ₃ ).

6b NMR (DMSO). ¹³ C: 103.7 (C-1; 77.6 (C-3 and C-5); 74.3 (C-2); 70.9 (C-4); 69.4

(C-α); 61.9 (C-6); 32.1 - 22.9 (CH ₂ ) _n ; 14.8 (CH ₃ ).

Example 4. Preparation of l-O-alkyl-3-O-n-butanoyl-D, L-glycerols Butla-c (type II). 1.10 mol of compound la-c are reacted with stirring at room temperature with 10.7 g (0.1 mol) of n-butanoyl chloride in the presence of 11.1 g (0.11 mol) of triethylamine in 150 mL of anhydrous toluene. After 10 h of reaction, the solution is filtered and the toluene is evaporated under reduced pressure. The Butla-c products are isolated in the pure state by chromatography on neutral silica (eluent hexane-acetone 90:10 v / v). Table 5.

Example 5. Preparation of 10-alkyl-5-On-butanoyl-2,3-O-isopropylidene-D, L-xylitols But2a-c and of 10-alkyl-5-On-butanoyl-D, L-xylitols But3a- c (type II). 0.10 mol of compounds 2ac is reacted with stirring at room temperature with 10.7 g (0.1 mol) of n-butanoyl chloride in the presence of 11.1 g (0.11 mol) of triethylamine in 150 mL of anhydrous toluene. After 15 hours of reaction, the solution is filtered and the toluene is evaporated under reduced pressure. The But2a-c products are isolated in the pure state by chromatography on neutral silica (eluent hexane-acetone 90:10 v / v).

Table 6.

A fraction of But2a-c products is deacetalized in water-ethanol 95 ° GL with 2 g of Amberlyst 15H ⁺ resin per gram of acetal But2a-c, at 50 ° C. After disappearance of the acetal, filtration and evaporation of the solvent, the But3a-c compounds are isolated in the pure state by chromatography on neutral silica (hexane-acetone 60:40 v / v).

Table 7.

EXAMPLE 6 Study of Acute Toxicity in Mice

Animal protocol The toxicity by single administration in standard mice (5 males and 5 females EOPS per dose to be tested; 4 weeks of age; weight of 20 ± 5 g, OF1 mouse from IFFA CREDO) was conducted in accordance with the prescriptions of the OECD Guideline 401, over a 14-day observation period. The mouse is not rationed in food or water. The animals are weighed at the entrance to study and then at 7 ^ιeme and I4 ^Leme day. The average daily weight gain or GMQ, making it possible to judge the state of general metabolic form, is calculated for the periods D ₀ to D ₇ and D ₇ to D ₁ , by dividing the weight gain of the mouse over the period by the number of days of observation. Preparation of the products The products tested are: la-c, 2a-c, 3a-c, But2a-c, But3a-c, 4a, b and 5a, b. These products dissolved in an atoxic solvent tested on a batch of control animals (physiological serum-DMSO 60:40 v / v, or pure DMSO), or injected without solvent for those who presented in the liquid or syrupy state , were administered intraperitoneally (IP) and orally (orally, PO). Experimental results The mortality observed at lg.kg ^"1 by administration IP and at 3 g.kg ^{" 1} by administration PO is reported in table 8. In general, mortality is low at the doses tested and the average daily gain (ADG) of surviving in particular between 8 and 14 ^ιeme ^lth day is close to that observed with the control.

Table 8. Acute toxicity in Mice and Average Daily Gain (GMQ) of survivors between D ₈ to D ₄ Intraperitoneal route (IP), 1 g- g 'over 14 days Oral route (PO), 2 g.kg ^"1 of 14 days

% GMQ mortality (gj ^"1 )% GMQ mortality (d- ¹ )

Product F M F + M F M F M F + M F M

Control * 0 0 0 0.94 ± 0.20 0.98 ± 0.20 0 0 0 0.91 ± 0.12 0.97 ± 0.20

DMSO 0 0 0 0.88 ± 0.20 0.90 ± 0.18 la 40 20 30 0.90 ± 0.17 0.96 ± 0.22 lb 40 20 30 0.82 ± 0.29 0.93 ± 0.34

2b 0 0 0 0.97 ± 0.31 0.95 ± 0.26

3a 0 20 10 0.94 ± 0.31 0.93 ± 0.25

3b 20 20 20 0.97 ± 0.24 0.97 ± 0.20

4b 0 20 10 0.91 ± 0.22 0.94 ± 0.17

5b 0 0 0 0.92 ± 0.19 0.98 ± 0.26

Butla 20 20 20 0.96 ± 0.09 0.99 ± 0.24 20 0 10 0.91 ± 0.10 0.97 ± 0.27

Butlb 0 20 10 0.97 ± 0.23 0.96 ± 0.19 0 0 0 0.94 ± 0.14 0.89 ± 0.08

Butlc 0 0 0 0.94 ± 0.24 1.04 ± 0.26 0 0 0 0.95 ± 0.21 0.92 ± 0.17

But2a 20 20 20 0.95 ± 0.15 0.91 ± 0.17 60 60 60 * 0.90 ± 0.28 0.97 ± 0.41

Goal2b 0 0 0 0.91 ± 0.18 0.98 ± 0.22 0 0 0 0.97 ± 0.20 1.02 ± 0.24

But2c 0 0 0 0.95 ± 0.12 0.94 ± 0.28 0 0 0 0.91 ± 0.06 0.98 ± 0.22

But3a 0 0 0 0.90 ± 0.08 0.94 ± 0.20 0 0 0 0.90 ± 0.10 0.91 ± 0.30

Goal3b 20 40 30 0.92 ± 0.14 0.92 ± 0.28 0 0 0 0.94 ± 0.20 0.94 ± 0.08

But3c 40 40 40 0.85 ± 0.21 0.91 ± 0.34 40 40 40 0.75 ± 0.29 0.86 ± 0.21 * LD50 = 2.70 ± 0.559 g.kg ^" 'in the female and 2.7 ± 0.55 g.kg ^"1 in male Example 7. Control of dermatological safety in the Albino Rabbit. The purpose of the study is to determine in the albino rabbit the irritant properties and / or the degree of corrosion caused by a product during its single cutaneous application.

General protocol The dermal irritation and / or corrosion following the cutaneous application of a product to the back of the Rabbit are evaluated by rating the reactions observed, according to a determined scale (below). Experimentation leads to the calculation of the primary skin irritation index. Irritation index Class 0.00 <I.I.C. <0.50 Non Irritant (N.I.) 0.50 <I.I.C. <2.00 Slightly Irritant (L.I.) 2.00 <I.I.C. <5.00 Moderately Irritating (M.I.) 5.00 <I.I.C. <8.00 Severely Irritant (S.I.)

The protocol implemented, in accordance with the prescriptions of the OECD Guideline No. 404 (May 12, 1981) and the Official Journal of the French Republic (February 21, 1982), makes it possible to completely assess the reversible or irreversible nature of observed effects. The study is conducted in the spirit of Good Laboratory Practices.

Preparation of animals and products Albino ELCO rabbits are used, 3 animals of each sex per product. Their weight is between 1.5 and 1.7 kg at the start of the test, which begins with a 3-day acclimatization. The animals are housed in cages with standardized dimensions. The excrement is eliminated automatically. The cages are placed in an air-conditioned pet store (18-20 ° C), maintained at a humidity between 45 and 65% relative humidity. The air is absolutely filtered and renewed 10 times per hour. The semi-artificial nycthemeral cycle is 12 hours of light and 12 hours of darkness. The UAR “maintenance” food (referenced ISO 9002) is used. City water is distributed at will. The product used is applied to the dorsal skin region. To do this, the day before application, the dorsal region of each rabbit is carefully shaved over 2 areas of 14 cm by 5 cm symmetrical with respect to the vertebral axis. Everyone's left flank of animals is left intact, while the right flank is scarified with vaccinostyle, by three lines of 2.5 cm long and spaced 0.5 cm apart. The adjacent surfaces of the untreated skin of each animal serve as a control for the test. The test substance is placed on a square of gauze, which are held in place and protected by sterile compresses and a non-occlusive dressing. Experimental results After 24 hours of contact and 72 hours of contact respectively, are evaluated the possible lesions which may have appeared on the scarified zones or not of each of the animals of the two sexes, according to a scale relating to 5 levels of erythema and 5 levels of 'edema. The primary skin irritation index (CII) is calculated by adding the edema and erythema scores for 3 rabbits, for the 2 test areas. The total of these two figures is divided by 12 (2 observation times x 2 zones x 3 rabbits). The results, reported in Table 9, show that the products are classified NON-IRRITATING to SLIGHTLY IRRITANT.

Table 9. Control of dermatological safety in the Albino Rabbit.

^a Not significant difference with the corresponding base ^b The protocol implemented complies with the prescriptions of the guideline n ° 404 of the OCDED of May 12, 1981. The calculation of the index of primary cutaneous irritation is carried out under the conditions provided for in the OJ of 21 February 1982. ^c Reminder of the classifications; 0.00 <IIC <0.50 non-irritating (NI) 0.50 <IIC <2.00 mildly irritating (LI) 2.00 <IIC <5.00 mildly irritating (MI) 5.00 <IIC <8, 00 severely irritating (SI) EXAMPLE 8 Antiproliferative Effect of Various Compounds Against Cancer Lines and of a Line of Human Keratinocytes with Type II Products: Cancer Lines: Line HT1080 (human fibrosarcoma; line established in 1972 from a human bone tumor), - Line EMT6 (murine bladder tumor; line supplied by Pr JDChapman, Fox Chase Cancer Institute, Philadelphia, USA). Human keratinocytes: - Line NCTC 2544 (Perry, VP, Sandford, KK, Hyatt, GW, Earle, WR Establishment of epithelial cells from human skin. J. Natl. Cancer Inst. (1957) 18: 707-717.

Figure 1 attached (Figure 1) illustrates the effect of various On-butanoyl-O-alkyl type II derivatives, at a concentration of 10 ^~ M, on the proliferation of HT 1080 cells, in comparison with sodium butyrate ( BuNa) at the same concentration (typical experience); influence of the length of the alkyl chain (experimental conditions: 48 hours of treatment with the products in DMEM medium supplemented with 5% fetal calf serum). The examination of Figure 1 (in the appendix) makes it possible to make several observations: 1 / Sodium butyrate, at the concentration of 10 ^"4 M, has no antiproliferative effect. This result is already known, butyrates do not being effective only above 10 ^" 3M, a concentration ten times higher. The concentration of 10 ^"4 M was chosen in view of preliminary experiments showing a notable efficacy of the claimed compounds, at this concentration, while the butyrates have no significant effect (results not shown). It can therefore be stated that the compounds butlb , but2b and but3b are at least 5 to 10 times more effective than sodium butyrate, a compound already used in the treatment of certain tumors. 2 / For the same substrate (1, 2 or 3), the length of the chain grafted alkyl is very critical for the antiproliferative effect, so an 8-carbon chain is almost completely ineffective regardless of the itol (butla, but2a, but3a). obtained with a chain with 12 carbon atoms (butlb, but2b, but3b), and decreases beyond (butlc, but2c). This observation strongly suggests that the studied compounds can, by an insertion at a precise level in the memb cell line, depending on the length of the alkyl chain, alter specific membrane processes. Indeed, a simple facilitating effect of the alkyl chain on cell penetration should not cause such a "critical" effect of the chain length to appear. These observations lead us to reconsider the potential mechanisms of action. Thus, it seems that in this concentration zone, the characteristics of the O-alkyl chain are much more important than the presence of the On-butanoyl group (butyrate itself having no effect at this concentration, other mechanisms should be considered). These observations gave rise to new experiments, carried out with the corresponding type I compounds, that is to say the O-alkyl compounds lacking an O-butanoyl group. Figure 2 (in the appendix) shows some examples of the results obtained, still on the human cancer line HT1080.

Figure 2 attached (Figure 2) illustrates the effect of various type I derivatives on the proliferation of HT1080 cells, in comparison with sodium butyrate (BuNa) and with the corresponding type II derivatives (concentration: 10 ^'4 M , treatment time 48h).

It can thus be seen that the three derivatives tested of type I above, that is to say the O-alkyl derivatives devoid of butyrate are at least as active as their type II analogs, which confirms that the antiproliferative effect n is not linked to the presence of the butanoyl group. From this observation, experiments were carried out with O-alkyl derivatives of type I, by varying the nature of the substrate (ose or itol) more widely. From the results presented in Figure 3 (in the appendix), we were able to conclude that, for the same length of the alkyl chain (Cι ₂ ), the order of effectiveness is as follows: α-L-Fucose>β-D-Glucose> β-D-Galactose and β-D-Galactose = D, L-Xylitol = D, L-Glycerol

Figure 3 appended (Figure 3) illustrates the effect of various type I derivatives with a Ci ₂ chain, on the viability of human tumor cells HT 1080. (concentration: 10 ^' M, treatment time: 48h).

For O-alkyl most active derivatives (chain Cι _2), the nature of the monosaccharide is not unimportant. Figure 4 (in the appendix) shows an example, in phase contrast optical microscopy, of the powerful cytotoxic effect of the fucose derivative having a C ₁₂ alkyl chain (4b). Figure 4 appended (Figure 4) illustrates the phase-shift contrast microscopy examination of cultures of human cancer cells (HT 1080 line) control or treated for 48 hours with the O-alkyl derivative of fucose (4b) at different concentrations. A: witnesses (not treated); B: 10 ^'5 M; C: 510 ^'5 M; D: 10 ^'4 M.

Finally, Figure 5 (in the appendix) shows an example of an experiment carried out with product 4b, showing the dose-dependence as well as the dependence of the cytotoxic effect on the treatment time, for the same concentration.

Figure 5 attached (Figure 5) illustrates the cytotoxic effect of product 4b on human tumor cells HT 1080, depending on the concentration (10 ^' M to 10 ^' M) and the treatment time (1 to 4 days: Jl at D4). OJ = controls (100%).

Example 9. Effects of compounds of type I and II on the viability of the tumor line EMT6 (murine bladder cancer). Figure 6 (in the appendix) presents only part of the results obtained.

Figure 6 attached (Figure 6) illustrates the effect of type I derivatives on the proliferation of EMT6 cells, in comparison with sodium butyrate (BuNa) and with the corresponding On-butanoyl-O-alkyl derivatives (type II) (butlb, but2b, but3b), at a concentration of 10 ^' M (treatment time: 48h).

It can be seen that, overall, the results concerning these compounds are fairly close to those obtained for the HT1080 line. However, there is a greater sensitivity to O-alkyl derivatives (type I), which suggests that the efficacy of the products may vary depending on the tumor line studied and perhaps depending on the differences in composition and physical characteristics of the membranes depending on the cell type considered. Differences in efficiency regarding the length of the alkyl chain were also found.

Example 10. Effects of Type II Compounds on the Human Keratinocyte Line

NCTC 2544.

Figures 7 and 8 (in the appendix) show some of the results obtained with the line (non-tumor) of human keratinocytes NCTC 2544, considered to be fairly representative of the proliferative layer (basal stratum) of the epidermis. We can see that, overall, the results go in the same direction as those presented for the tumor lines, but with slight differences, in particular a maximum efficiency for but2b, which confirms the existence of differences in sensitivity depending on the cell type. There are also significant differences depending on the length of the alkyl chain (C ₈ , C ₁₂ or Cι ₆ ). Finally, the efficiency of the O-alkyl derivatives (Figure 8; in the appendix) is roughly equivalent to that of the O-n-butanoyl-O-alkyl derivatives (Figure 7; in the appendix).

Figure 7 attached (Figure 7) illustrates the effect of various type II derivatives on the viability of human keratinocytes NCTC 2544 in culture (concentration: 10 ^' M, treatment time: 48h). (butla, but3a and but2a: C alkyl chain; butlb, but3b and but2b: C12; butlc, but3c and but2c: Cm).

The attached figure (Figure 8) illustrates the effect of some type II derivatives with a Cn alkyl chain (butlb, but3b, but2b), compared with that of corresponding type I derivatives (1b, 3b and 2b). (concentrations 10 ^' M, duration of treatment: 48h).

Example 11. Potentiation of the photocytotoxic effect of Photofrin II ^R by the product of type I, 4b.

This example concerns photochemotherapy of tumors (PCT). This method, in full development, is based on the photosensitized generation of activated oxygen species, leading to the destruction of tumor cells and to the vascular supply of the tumor (Dougherty, TJ 1989, Oncology, 3, 67; Dougherty, TJ , 1993, Photochem. Photobiol. 58, 895; Pass, HI & al, 1993, J. Natl. Cancer. Inst, 85, 443). The main advantage of this method is selectivity, since only the illuminated area, i.e. the tumor (illuminated externally for skin tumors, or endoscopically for tumors of hollow organs ) is concerned with the photocytotoxic effect. Despite its advantages, MDT still faces certain obstacles: poorly vascularized tumors (the effectiveness of treatment depends on the oxygen concentration of the tissues: Chapman, JD & al. 1991, Radiât. Res., 126, 73), or certain cases of resistance of various origins (re-excretion of the photosensitizer etc.). To overcome these problems, potentiation strategies combining a photosensitizer with a non-photosensitizer molecule increasing the cellular response, have been developed (Biade, S. & al, 1993, Photochem. Photobiol., 57, 371; Biade, S. & al., 1993, Cell Pharmacol., 1, 37; Berg, K. & al., 1998, Biochim. Biophys. Acta, 1370. 317). In this example, we present results showing a potentiation of the photocytotoxic effect of Photofrin II ^R , photosensitizer already used in human therapy (phase III) and one of the type I derivatives described above, the L-fucose derivative, 4b. The HT1080 tumor cells were pretreated for 48 h with 4b at non-cytotoxic doses (10 ^" and 5 10 ^" M). According to FIG. 9, we show that a 48h pretreatment with one of the 0-alkyl derivatives previously described, 4b, at concentrations having no specific cytotoxic effect (10 ^" and 5x10 ^" M), potentiates a factor close to 2 the photocytotoxic effect of Photofrin II ^R , a photosensitizer already used in human therapy for the photochemotherapy of various tumors ((Dougherty, TJ 1989, Oncology, 3, 67).

Figure 9 attached (Figure 9) illustrates the potentiation of the photocytotoxic effect of Photofrin Ir (IPSEN Biotech) by non-cytotoxic concentrations of the compound of type I, 4b. HT 1080 cells are pretreated for 48 hours with low concentrations of 4b (10 ^' or 5 10 ^' M) which do not affect cell viability. The

Photofrin if is then added (final concentration in the culture medium: lμg.mL) and the incubation extended for 24 hours. The cells are then washed, then undergo an illumination dose of 0.3J.cm in UVA (Vilber Lourmat table TFL 35). (T = control; FI = 4b only; P2 = Photofrin if * only; P2F1 = 4b + Photofrin if ¹ ).

Figure 10 (in the appendix) shows the appearance, in phase contrast optical microscopy, of the cells thus treated (for a 4b concentration of 10 ^"6 M). The appended figure 10 (Figure 10) illustrates the examination in optical microscopy of HT1080 cells pretreated for 48 h with FI at 10 ^'6 M then with Photofrin if ¹ at lμg.mL ^' for 24h A: Photofrin II alone, no illumination; B: 4b alone, no illumination (it is noted that there is no effect of 4b on the cells at this concentration); C: Photofrin II alone, UVA 0.3 J. cm ² ; D: Photofrin II + 4b, UVA 0.3 J. cm ² .

By comparing D (Photofrin II + 4b) and C (Photofrin II alone), a marked increase in cellular damage after illumination is observed. The appearance of damaged cells evokes a process of apoptosis (retraction of cell coars, refraction, many very dense debris).

Example 12 Curative Effects of Type II Compounds on Psoriasis The compound 3-O-n-butanoyl-1-O-n-hexadecylglycerol (Butlc) was tested in a hospital environment on a male patient suffering from severe psoriasis. The product was applied in solution of 0.1% (m / m) in ethyl oleate. It was checked beforehand on 100 healthy subjects that the application on the right arm of this solution and on the left arm of ethyl oleate alone for three months did not cause any skin reaction. The allergenicity test was carried out on the Rabbit and revealed no reaction. In view of this result, the patient was treated by applying the above solution daily to the patient's legs for two and a half months. Figure 11a (in the appendix) shows the patient's condition before the treatment and Figure 11b (in the appendix) shows the disappearance of the lesions initially present on this same patient fifteen days after the end of the treatment. No recurrence was noted in the following six months.

Figures 11a and 11b attached (Figure 11a and Figure 11b) (a and b). illustrates the effect of the Butlc product in daily application for two and a half months on a patient with acute psoriasis. (Figure 11a) Patient before treatment. (Figure 11b) Patient fifteen days after the end of treatment.

Claims

Claims:

1- New saccharide and itol derivatives with an O-alkyl group and corresponding to the general formula I

in which Su represents a cyclic or unprotected saccharide derivative, protected or not, or alternatively an itol derivative, cyclic or not, protected or unprotected, in which R represents an n-alkyl or n-alkenyl chain having 8 to 18 atoms carbon, in which P represents a group of atoms linked to the oxygen atom of the hydroxyl group and forming, via the latter, and with Sub, an ether function, or even forming an ester function, or even forming an acetal of dioxolane type, in which i represents the number of hydroxylated sites not carrying R as defined above, these hydroxylated groups being able to be all (j = 0) or partly free (0 <j <i) or even protected in the form of groups OP with P as defined above, in which j represents the number of hydroxyl groups protected in the form of OP groups, the j groups P can be identical or not. Excluded from this claim are the compounds derivatives of the 3-On-alkyl-1,2: 5,6-di-O-isopropylidene-α-D-glucofuranose, 3-On-alkyl-1,2-O-isopropylidene type. -α- D-glucofuranose, 3-On-alkyl-α-D-glucopyranose and 3-On-alkyl-β-D-glucopyranose. Also excluded from this claim are derivatives of D-xylose, D-xylitol and L-xylitol carrying an n-alkyl chain and not carrying an On-butanoyl group. Also excluded from this claim are the α-D-glucopyranoside derivatives of n-alkyl protected or not and the β-D-glucopyranosides of n-alkyl protected or not bearing no butanoyl group. Also excluded from this claim are the α-D-galacopyranosides of protected or unprotected n-alkyl and the β-D-galacopyranosides of protected or unprotected n-alkyl not bearing a butanoyl group. Also excluded from this claim are glycerol derivatives not carrying a butanoyl group. Also excluded from this claim are compounds not bearing butanoyl group and of which at least one of the P groups is represented by an acetyl group.

2- New saccharide and itol derivatives according to claim 1 having an O-alkyl group and an O-n-butanoyl group corresponding to the general formula

II

II in which Su represents a cyclic or unprotected saccharide derivative, protected or not, or alternatively an itol derivative, cyclic or not, protected or not, in which R represents an n-alkyl or n-alkenyl chain having 8 to 18 atoms carbon, in which P represents a group of atoms linked to the oxygen atom of the hydroxyl group and forming, via the latter, and with Sub, an ether function, or even forming an ester function, or even forming a dioxolane-type acetal, in which i represents the number of hydroxylated sites not carrying R as defined above, these hydroxylated groups being able to be all (j = 0) or partly free (0 <j <i) or even protected in the form of OP groups with P as defined above, in which j represents the number of protected hydroxyl groups in the form of OP groups, one P group of which is represented by the butanoyl group.

3 - Compounds, derivatives of saccharides and itols according to claims 1 and 2, in which the substrate Sub is chosen for example from hexose, pentose, deoxyhexose, or even an itol derivative, cyclic or not, protected or not corresponding for example glycerol.

4- Compounds, derivatives of saccharides and itols according to claims 1 and 2, in which P is preferably chosen from benzyl or trityl, acetyl, benzoyl groups, or else forms an acetal of dioxolane type, preferably isopropylidene or benzylidene . 5- octyl α-L-fucopyranoside

6- dodecyl α-L-fucopyranoside 7- hexadecyl α-L-fucopyranoside

8- 3-O-n-butanoyl- 1 -O-n-octyl-D, L-glycerol

9- 3-O-n-butanoyl-1-O-n-dodecyl-D, L-glycerol

10- 3 -O-n-butanoyl- 1 -O-n-hexadecyl-D, L- glycerol '

11- 5-O-n-butanoyl-2,3-O-isopropylidene-1-O-n-octyl-D, L-xylitol and 5-O-n-butanoyl-1- O-n-octyl-D, L-xylitol

12- 5-O-n-butanoyl-1-O-n-dodecyl-2,3-O-isopropylidene-D, L-xylitol and 1-O-n-dodecyl- 5-O-n-butanoyl-D, L-xylitol

13- 5-O-n-butanoyl-1-O-n-hexadecyl-2,3-O-isopropylidene-D, L-xylitol and 1-O-n-hexadecyl-5-O-n-butanoyl-D, L-xylitol

14- Use of one of the compounds, derived from saccharides and itols, having an O-alkyl group and corresponding to the general formula I

in which Su represents a cyclic or unprotected saccharide derivative, protected or not, or alternatively an itol derivative, cyclic or not, protected or unprotected, in which R represents an n-alkyl or n-alkenyl chain having 8 to 18 atoms carbon, in which P represents a group of atoms linked to the oxygen atom of the hydroxyl group and forming via it, and with Sub, an ether function, or also forming an ester function, or even forming an acetal of dioxolane type, in which i represents the number of hydroxylated sites not carrying R as defined above, these hydroxylated groups being able to be all (j = 0) or partly free (0 <j <i) or even protected in the form of OP groups with P as defined above, in which j represents the number of hydroxyl groups protected in the form of OP groups, the j groups P can be identical or not, a group P can be represented by the butano group yle, for the preparation of anti-cancer drugs locally (skin tumors) or general.

15- Use of one of the compounds, derived from saccharides and itols, having an O-alkyl group and corresponding to the general formula I

in which Su represents a cyclic or unprotected saccharide derivative, protected or not, or alternatively an itol derivative, cyclic or not, protected or unprotected, in which R represents an n-alkyl or n-alkenyl chain having 8 to 18 atoms carbon, in which P represents a group of atoms linked to the oxygen atom of the hydroxyl group and forming, via the latter, and with Sub, an ether function, or even forming an ester function, or even forming an acetal of dioxolane type, in which i represents the number of hydroxylated sites not carrying R as defined above, these hydroxylated groups being able to be all (j = 0) or partly free (0 <j <i) or even protected in the form of groups OP with P as defined above, in which j represents the number of hydroxyl groups protected in the form of OP groups, the j groups P can be identical or not, a group P can be represented by the butan group oyle, for the preparation of antiproliferative drugs in benign tumors, in particular benign skin tumors.

16- Use of one of the compounds, derived from saccharides and itols, having an O-alkyl group and corresponding to the general formula I

in which Su represents a cyclic or unprotected saccharide derivative, protected or not, or alternatively an itol derivative, cyclic or not, protected or unprotected, in which R represents an n-alkyl or n-alkenyl chain having 8 to 18 atoms carbon, in which P represents a group of atoms linked to the oxygen atom of the hydroxyl group and forming, via the latter, and with Sub, an ether function, or also forming an ester function, or alternatively forming an acetal of dioxolane type, in which i represents the number of hydroxylated sites not carrying R as defined above, these groups hydroxylated which may be all (j = 0) or partly free (0 <j <i) or else protected in the form of OP groups with P as defined above, in which j represents the number of hydroxylated groups protected in the form of OP groups, the j groups P can be identical or not, a group P which can be represented by the butanoyl group, for the preparation of antiproliferative drugs in non-tumor proliferative pathologies, in particular psoriasis.

17- Use of one of the compounds, derived from saccharides and itols, having an O-alkyl group and corresponding to the general formula I

I in which Su represents a cyclic or unprotected saccharide derivative, protected or not, or also an itol derivative, cyclic or not, protected or unprotected, in which R represents an n-alkyl or n-alkenyl chain having 8 to 18 atoms carbon, in which P represents a group of atoms linked to the oxygen atom of the hydroxyl group and forming, via the latter, and with Sub, an ether function, or also forming an ester function, or even forming a dioxolane-type acetal, in which i represents the number of hydroxylated sites not carrying R as defined above, these hydroxylated groups being able to be all (j = 0) or partly free (0 <j <i) or even protected in the form of OP groups with P as defined above, in which j represents the number of hydroxylated groups protected in the form of OP groups, the j P groups can be identical or not, a P group can be represented by the butan group oyle, for the preparation of potentiating agents making it possible to improve the effectiveness of anti-tumor photochemotherapy. 18- Use of one of the compounds, derived from saccharides and from itols, having an O-alkyl group and corresponding to the general formula I

I in which Su represents a cyclic or unprotected saccharide derivative, protected or not, or also an itol derivative, cyclic or not, protected or unprotected, in which R represents an n-alkyl or n-alkenyl chain having 8 to 18 atoms carbon, in which P represents a group of atoms linked to the oxygen atom of the hydroxyl group and forming, via the latter, and with Sub, an ether function, or also forming an ester function, or even forming a dioxolane-type acetal, in which i represents the number of hydroxylated sites not carrying R as defined above, these hydroxylated groups being able to be all (] -0) or partly free (0 <j <i) or even protected in the form of OP groups with P as defined above, in which j represents the number of hydroxylated groups protected in the form of OP groups, the j P groups may or may not be identical, a P group may be represented by the goal group anoyl, for the preparation of drugs included in arterial prostheses, in particular for the purpose of reducing cell proliferation.