WO1996016957A1 - 3,4-dihydro-2,5,7,8-tetramethyl-benzopyran-6-ol derivatives for use as drugs - Google Patents

Abstract

Compounds of general formula (I), as defined in the description, salts, hydrates, solvates and therapeutically acceptable prodrugs thereof, as well as racemic forms and enantiomers thereof, are disclosed. A method for preparing the compounds, and pharmaceutical compositions containing said compounds as the active principle for treating and/or preventing acute or chronic inflammatory diseases, are also disclosed.

Description

 Derivatives of 3-4 dihydro 2, 5, 7, 8 tetramethyl-benzopyran-6-ol useful as medicaments

The present invention relates to new derivatives of 3,4 dihydro-2,5,7,8 tetramethyl-benzopyran-6-ol, their manufacturing process, the pharmaceutical compositions containing them and their use as medicaments. The generation of highly reactive oxygen metabolites is an essential feature of many components of normal metabolism (generation of energy metabolism by the mitochondrial respiration chain, detoxification of xenobiotics by cytochromes, destruction of microorganisms by phagocytosis, and even ovulation and fertilization).

Despite the existence of powerful endogenous antioxidant defense systems, the flow of locally generated oxidizing species can frequently exceed the body's natural defenses and lead to tissue destruction due to the peroxidation of the membrane lipids of cells and organelles, denaturation of structural and functional (enzymatic) proteins, radical cleavage of DNA or RNA strands, and alteration of the polysaccharide constituents of interstitial tissue and basement membranes. As a result, this local toxicity of free radicals and other toxic derivatives of oxygen constitutes a path common to numerous pathologies such as damage caused by radiotherapy, hyperoxygenation syndromes (for example neonatal bronchopulmonary dysplasia), inflammatory reactions (cf. GB Buckley, Surgery, 112, 479, 1993), autoimmune diseases, toxic damage during the post-ischemic reperfusion phases (JY Arigou et al., Arch. Mal. Coeur, 86, 105-109 , 1993; N. Kaul et al. J. Pharmacol. Tox. Methods, 2fl, 55-67, 1993) reperfusion of organs including transplanted organs, carcinogenesis (cf. H. Sies, Ang. Chem. Int. Ed. ., 25, 1058-1071, 1986), atherosclerosis (cf. D. Steinberg, New England J. Med. 22Û, 915-924, 1989; M. Aviram, Atherosclerosis, 28, 1-9, 1993) and aging processes at the cellular level (cf. E. Stadtman, Science, 252, 1220, 1992; B. Ames and M. Shigenagu, Ann. NY Acad. Sci ences, 85, 1993; PNAS, 9_A, 7915, 1993) or neuronal (C. Olanow, TINS, 1 £, 439, 1993; C. Smith et al. Ann. NY Acad. Sciences, 110, 1993). Numerous pharmacological and clinical studies suggest that certain derivatives with antioxidant properties may play a beneficial role in the treatment and / or prevention of the pathologies mentioned above. As examples include probucol, vitamin C, β-carotene, flavonoids such as quercetin or ebselen, (H. Sies, "Oxidative stress", Académie Press, 1991; A. Ong, L. Packer, "Lipid-soluble antioxidants: biochemistry and clinical applications", Birkhauser, 1992; B. Halliwell, Drugs, 42, 569, 1991; M. Santrucek, J. Krepelka, Drugs of the future, 13, 974, 1988), as well as 2H-1-benzopyrans derivatives such as vitamin E or tocopherols, trolox, tocotrienols, derivatives MDL 72720, 74366 and 74405 (Free Radical Biol. Med., 15, 209, 1993; Eur. J. Pharmacol. 122, 241, 1990 and 122, 383, 1991, Cardiovasc. Res., 28, 295, 1994), the derivative U-78518 (J. Appl. Physiol. 24, 2155, 1993), the derivative U-83836 [Clin. Res. 4fl (4): 828A-830A (1992); 41 (2): 327A, 387A (1993)] or the derivative WIN 62079 (PJ. Silver, Drug. Dev. Res. 22, 45, 1992). The Applicant has now discovered new derivatives of 3,4 dihydro-2,5,7,8-tetramethyl-2H-1-benzopyran-6-ol which have interesting pharmacological and therapeutic properties since these derivatives behave like antioxidants capable protect biological systems such as tissues, membranes, cells, structural and / or functional proteins and lipids from aggression due to free radicals and other toxic oxygen derivatives. The very good efficacy (greater than vitamin E) of the compounds of the invention as antioxidants, (in particular their capacity to protect human LDL from oxidative modifications induced by copper or endothelial cells and their capacity to trapping hydroxyl radicals) associated with their stability and low toxicity, makes them valuable for the treatment of pathologies in which peroxidation plays an initiating and / or aggravating role. In addition to the publications cited above, the prior state of the art in this field is illustrated in particular by:

US Patents 5,135,945 and EP 369,082 which describe tocopherol analogs having an alkylamino chain in position 2 on the 2H-1-benzopyran ring. The patents EP 293 078, EP 358 676, EP 487 510, US 5120843 and WO 8808424 which describe tocopherol analogs having a methyl amine (such as a substituted piperazine) in position 2 on the 2H-1-benzopyran ring. European patent application EP 587 499-A1 which describes thioamides derived from benzopyrans. The publication J. Org. Chem. 58, 3521 in 1993 which describes the antioxidant activity of aromatic ethers derived from benzopyran. The structural modifications made to the side chain of tocopherols have led to the derivatives of the present invention, which differ from the closest derivatives of the prior art not only by their original chemical structure but also by their biological profile and their therapeutic potential.

The subject of the present invention is new derivatives of 3,4-dihydro-2,5,7,8-tetramethyl-benzopyran-6-ol corresponding to the general formula (I)

in which n represents an integer between 1 and 10 R represents CH2OR1, CONR1R2, CH2NR1R2 or A in which Ri, R2, identical or different, each represent a hydrogen, a linear or branched alkyl radical comprising from 1 to 20 carbon atoms which can itself contain one or more unsaturations such as double or triple bonds and can be substituted by an aromatic residue (such as a phenyl, a pyridine or a benzopyran variously substituted) by a halogen (chlorine, bromine or fluorine), by an alkoxy, (OR4) by a thiother (SR4), by a silane (SÎR4R5R6) or by an amine (NR4R5) in which R4, R5 and R _Ô represent an alkyl or aryl radical.

R and R2, when attached to the same nitrogen atom, can also form a variously functionalized ring such as a 3.8 diazo-l-oxa-2-oxaspiro [4,5] decane.

Ar represents an aromatic radical such as a phenyl which can be variously substituted in various positions by a halogen (chlorine, bromine, fluorine or iodine), an amine (NR1R2), an alkoxy (OR), a thiother (SRi), a radical linear or branched alkyl comprising from 1 to 10 carbon atoms in a straight or branched chain which can itself be substituted by an alkoxy (ORi), an amine (NR1R2), or a thioether (SRi) and moreover, Ri and R2 can also form a cycle with the nitrogen to which they are attached. The compounds of formula (I) containing one or more asymmetric centers have isomeric forms. The racemates and then the enantiomers of these compounds are also part of this invention.

The invention also includes the salts, solvates (for example hydrates) and bioprecursors of these compounds acceptable for therapeutic use. The expression bioprecursors as used in the present invention applies to compounds which, when administered to an animal or to a human being, are converted in the organism into a compound of formula (I). It is thus and by way of nonlimiting example that derivatives of formula la

in which P (for example an acyl, a phosphoryl or a glycosyl) represents a bio-labile group (defined so that this compound la can be transformed into corresponding compound I within a living organism) must also be considered as part of the present invention.

The compounds of the present invention are generally prepared by condensation of an intermediate of formula (II)

in which P represents a protecting group for a phenol such as a methoxymethyl ether, a benzyloxymethyl ether, a methoxyethoxymethyl ether, a tetrahydropyranyl ether or alternatively a 2- (trimethyl-silyl) ethoxy methyl ether, with an appropriate electrophile which will depend on the nature of n and R: In the case of derivatives of general formula (I) in which R represents Aτ \ defined as above, a particularly appreciated synthesis method consists in condensing an intermediate of formula (II) with an electrophile of formula (III)

Y - (CH ₂ ) _n - Ari (III) in which Y represents a leaving group such as a halogen (bromine, chlorine or iodine) a mesylate, a tosylate or a triflate in the presence of an organic or inorganic base such as NaH, KH, ΦuOK, in an anhydrous polar solvent such as THF, DMF, DMSO ^or BuOH at a temperature between 0 and 60 ° C, followed by deprotection of the thus formed intermediate using appropriate methods depending of the nature of P and which are described in "Protective Groups in Organic Synthesis", TW Greene, P. W Wutz, J. Wiley & Sons, NY, 1991. Thus when P represents a 2- (trimethylsilyl) group ) -ethoxy methyl ether, a particularly preferred method consists in using sulfuric acid diluted in a methanol-THF mixture to carry out this transformation. In the particular case of derivatives of general formula (I) in which n = 1 and R represents an aromatic residue being substituted (in ortho, meta or para position) by a residue -CH2OR1 or -CH2NR1R2, an alternative synthesis method more particularly apreciated consists in first condensing the alcohol of formula (II) with a dibromo derivative of formula (TV)

in which the CH2Br groups are in various relative positions (ortho, meta or para) on the aromatic to which they are attached in the presence of NaH, in THF or DMF at a temperature between - 20 ° C and 0 ° C. This reaction leads to the formation of the intermediate (V) and of the product (VI) which after deprotection leads to the product VII which is also part of the present invention. Control of the experimental conditions (cf. examples x and y) makes it possible to promote the formation of one or the other product.

VI: P represents a protecting group

VII: P = H

The products of general formula (I) in which n = 1 and Ari represents an aromatic substituted by a CH2OR1 group are prepared by condensation of an alcohol of formula RiOH with the intermediate of formula (V) in the presence of a base such as NaH in a solvent such as THF, DMF or DMSO at a temperature between - 15 ° and 20 ° C, followed by deprotection of the phenol function as indicated above. The products of general formula (I) in which n = 1 and Ari represents an aromatic substituted by a group

CH2NR1R2 are prepared by condensation of an amine (R R2NH) with an intermediate of formula (V) in the presence of a base such as K2CO3, Na2CO3,

CS2CO3, DBU, DIPEA in the possible presence of Kl or BU4NI in a polar anhydrous solvent such as methyl ethyl atone, DMF, THF or DME at a temperature between 0 ° C and 60 ° C, followed by deprotection of the phenol as previously indicated.

The derivatives of formula (I) in which R represents CH2-OR1 are generally prepared from an ester of formula (VIII)

(in which P and R4 are described as above and n represents an integer between 1 and 10) obtained by condensation of an intermediate of formula II with an ester of formula (IX)

Y- (CH ₂ ) n ^

0-R „

(IX)

in which n, Y and R4 are described as above, in the presence of a base such as NaH, KH or ¹ BuOH in an anhydrous polar solvent such as DMSO, DMF or uOK.

The transformation of the esters of formula VIII into compounds of formula I in which R represents CH2-OR implements the reduction of the ester function to give the alcohol of formula (X) using reducers well known for this type of transformation ( such as for example lithium aluminum hydride in ether or THF) followed by condensation

of this alcohol (X) with an appropriate electrophile of formula Ri-Y in which n, Ri and Y are defined as above. The derivatives of formula (I) in which R represents ORi are finally isolated after deprotection of the phenol as indicated above. The compounds of formula (I) in which R represents -C (O) NRιR2 are prepared in 2 steps from an intermediate of formula (VIII) in which P, n and R4 are defined as above. This intermediate is first hydrolyzed to acid medium (a particularly preferred method for doing this consists in using an intermediate of formula (VIII) in which R is ^t_ butyl and in treating it with trifluoroacetic acid in dichloromethane) which leads to the simultaneous deprotection of phenol and product formation (XI)

qui sera lui-même transformé directement en composé de formule (I) dans laquelle R représente CONR1R2 après condensation avec une amine R1R2NH dans les conditions bien connues pour passer d'un acide à une amide telles que par exemple l'utilisation de carbonyl dimidazole, de pyBOP, de HOBT, de DCC en présence de bases diverses telles que la triéthylamine ou la N-méthyl morpholine dans un solvant polaire aprotique anhydre tel que le dichlorométhane, le THF ou le DME. Les composés de formule (I) dans laquelle R représente CH2NR1R2 sont préparés par condensation d'une amine R1R2NH avec un électrophile de formule (XII) dans laquelle P, n et Y sont définis comme précédemment,

which will itself be directly transformed into a compound of formula (I) in which R represents CONR1R2 after condensation with an amine R1R2NH under the conditions well known for passing from an acid to an amide such as for example the use of carbonyl dimidazole, pyBOP, HOBT, DCC in the presence of various bases such as triethylamine or N-methyl morpholine in an anhydrous aprotic polar solvent such as dichloromethane, THF or DME. The compounds of formula (I) in which R represents CH2NR1R2 are prepared by condensation of an amine R1R2NH with an electrophile of formula (XII) in which P, n and Y are defined as above,

in the presence of a base such as K2CO3, CS2CO3, DIPEA and optionally in the presence of an iodide such as K1 or BU4NI, followed by deprotection of the phenol as indicated above. The intermediates of formula (XII) are obtained from the alcohols of formula (X) by methods and techniques well known to those skilled in the art for transforming an alcohol into a leaving group, such as the use of SOCI2 or POCI3 for the preparation of products of formula XII in which Y = Cl, the use of PBr3 or Br2PCl3 for the preparation of derivatives of formula XII in which Y = Br, the use of PI3 or P2I4 for the preparation of derivatives of formula XII in which Y = I, the use of tosyl chloride for the formation of derivatives of formula XII in which Y represents a tosylate, the use of mesyl chloride for the preparation of products of formula (XII) in which Y represents a mesylate or also the use of triflic anhydride for the preparation of products of formula XII in which Y represents a triflate. These various transformations are generally carried out in an anhydrous aprotic polar solvent such as dichloromethane, dichloroethane or even THF, in the optional presence of a base such as a tertiary amine or a pyridine at a temperature of between -15 ° C. and 40 ° C. The intermediate alcohols of general formula (II) are prepared from the esters of 6-hydroxy-2,5,7,8-tetramethyl-chτoman-2-carboxylic acid of general formula (XIII)

in which R4 is defined as above, by initial introduction of the protective group on the phenol function according to the methods and techniques described in "Protective Groups in Organic Synthesis" by T.W. Greene and P. Wutz, J. Wiley & Sons, NY (1992) chosen according to the nature of P. The ester function of the intermediate thus obtained is then reduced to alcohol (formula II) by reaction with a lithium and aluminum hydride or a borane under conditions well known to those skilled in the art for this type of transformation.

The compounds of the present invention all have at least one asymmetric center located in position 2 on the pyrannic cycle. The preparation of the derivatives of formula (I) in the form of the pure isomers (R or S) at this asymmetric center (whether they are enantiomers or epimers) is carried out by substituting the racemic product (XIII) by its R enantiomer or by its S enantiomer for the preparation of the compounds of formula (I) by following the procedures described above and this, for each isomer. The pure enantiomers (R and S) of the esters of formula (XIII) are prepared by esterification of the corresponding optically pure acids (for example, the ethyl esters R and S of formula XIII which are particularly preferred for the preparation of the compounds of the present invention are obtained by reaction of the acids optically then R and S corresponding with ethanol in the presence of sulfuric acid) which are themselves accessible according to the methods described (for example J. Scott et al., J. Am. Oil Chem. Soc. 51, 200, 1974 and N. Cohen et al. J. Org. Chem. 4-6, 2445, 1981) . Also to be considered as an integral part of the present invention all the methods which make it possible to transform a derivative of formula (I) into another derivative of formula (I) by techniques and methods well known to those skilled in the art . Thus, and by way of example, the derivatives of formula (I) in which R represents CH2-NR1R2 can be prepared from a derivative of formula (I) in which R represents CONR1R2, by reduction to using agents derived from aluminum or boron such as lithium aluminum hydride in ether or THF.

When it is desired to isolate a compound of the present invention comprising basic nitrogen in the form of a salt, for example a salt by addition with an acid, this can be achieved by treating the free base of formula (I) with an appropriate acid of preferably in equivalent quantity. When the methods described above for preparing the compounds of the invention give mixtures of stereoisomers (in particular diastereoisomers) these isomers can be separated by conventional methods such as preparative chromatography. It will be understood that in certain reactions or sequences of chemical reactions which lead to the preparation of compounds of general formula (I) it is necessary or desirable to protect sensitive groups or functions in the synthesis intermediates in order to avoid undesirable side reactions. This can be achieved by the use of conventional protective groups such as those described in "Protective Groups in Organic Synthesis", TW Greene, J. Wiley & Jones, 1981 and "Protecting groups" by PJ Kocienski, Thieme Verlag, 1994. The adequate protective groups will therefore be introduced and then removed at the level of the most suitable synthetic intermediates for this and using the methods and techniques described in the references cited above. The following examples illustrate the invention without, however, limiting its scope. EXAMPLE 1 2 - [(benzyloxy) -methyl] -3,4-dihydro-2,5,7,8-tetramethyl-2H-1-benzopyran-6-ol 1

To a solution of Trolox® (20 g; 80 mmol) in absolute ethanol (800 ml) is added concentrated sulfuric acid (6 ml; 86 mmol; 1 J eq.) Then the resulting medium is stirred for 10 hours at reflux. The solvent is then evaporated and the solid obtained recrystallized from isopropanol to provide the expected ester in the form of a beige powder (16 g; 70%). m. p. = 120 ° C.

To a solution of ester obtained above (10 g; 36 mmol) in dichloromethane (70 ml) and maintained at 0 ° C. is added successively diisopropylethylamine (24.6 ml; 144 mmol; 4 eq.) Of N, N'- dimethylaminopyridine (220 mg; 1.8 mmol; 5%) and chloro-methyl-ethoxy-trimethylsilane (9.5 ml; 54 mmol; 1.5 eq.). The medium is then heated at reflux for 5 hours then cooled to room temperature, diluted in CH2Cl2 and washed with water until neutral pH. The organic phase is then dried over MgSO4, filtered and evaporated to give {3,4-dihydro-6 - [(2-trimethylsilylethoxy) -methoxy] -2.5, 7.8-tetramethyl-2H-1- ethyl benzopyran-2-yl} -carboxylate in the form of an orange oil sufficiently pure to be used without purification (14 g; ~ 100%).

The oil obtained previously (14 g; 34.3 mmol) is diluted in THF (300 ml) and brought to -30 ° C. The lithium aluminum hydride (1 M in THF; 34.3 mmol; 34.3 ml; 1 eq.) Is then added dropwise, then the temperature of the medium is brought back to 0 ° C. After one hour of reaction, the solution is hydrolyzed at 0 ° C with wet Na2SO4 then the resulting precipitate is filtered through celite and the solvent evaporated. The oil obtained is purified by "flash chromatography" on a silica column (50/50 Petroleum Ether / Ethyl Ether) to yield {3,4-dihydro-6 - [(2-irimethylsilyl-elhoxy) in the majority fraction. - methoxy] -2.5, 7.8-tetramethyl-2H-1-benzopyran-2-yl} -methanol Ha (P =

Me ₃ SiCH2CH ₂ OCH2) (9.8g; 78%). RN (CDCI3, 200 MHz, ppm): 4.96 (s, 2H); 3.85 (m, 2H); 3.66-3.48 (m, 2H); 2.59 (t, 6.9 Hz, 2H); 2J5 (s, 3H); 2.08 (s, 3H); 2.02 (s, 3H); 2.00-1.60 (m, 2H); 1.30 (m, 3H) .; 1.03-0.92 (m, 2H); 0.02 (s, 2H)

To a solution of ΝaH (55 mg; 1.75 mmol; 1.25 eq.) In 3 ml of DMF is added at 0 ° C the alcohol Ha (400 mg; 1.4 mmol) in 2 ml of DMF then the resulting medium is stirred 1 hour at room temperature. Benzyl bromide (250 μl; 2.8 mmol; 2 eq.) Is then added at 0 ° C and the reaction continued for 20 hours at room temperature. The medium is then diluted in ether, washed with water and then a saturated solution of ΝaCl. The organic phases are dried over MgSO4, filtered and evaporated. The oil obtained is purified by "flash chromatography" on a silica column (95/5 Petroleum Ether / Ethyl Ether) to provide the expected compound. Elementary Analysis for C27H39θ4Si Calculated: C: 71.16; H: 8.63 Found: C: 71.12; H: 8.68 To a solution of the silylated compound obtained above (334 mg; 0.73 mmol) in a mixture of THF (~ 12 ml) and MeOH (~ 12 ml) is added sulfuric acid (47 μl; 1 eq.) in solution in methanol (12 ml). The medium is stirred for 3 hours at room temperature and then neutralized with a saturated sodium bicarbonate solution. Part of the volatiles is evaporated and the resulting solution is diluted in water and extracted with ethyl acetate. The organic phases are dried over MgSθ4, filtered and evaporated. The oil obtained is purified by "flash chromatography" on a silica column (75/25 Petroleum Ether / Ethyl Ether) to provide product 1 (183 mg; 77%). NMR (CDCI3, 200 MHz, ppm): 7.40-7.20 (m, 5H); 4.60 (A ₂ B ₂ , 2H); 4.20 (br m, 1H); 3.48 (A ₂ B ₂ , 2H); 2.59 (t, 6.9 Hz, 2H); 2J5 (s, 3H); 2J0 (s, 6H); 2.08-1.92 (m, 1H); 1.85-1.68 (m, 1H); 1.30 (m, 3H). Elementary Analysis for C21H26O3 Calculated: C: 77.51; H: 7.74 Found: C: 77.01; H: 7.93 EXAMPLE 2

Synthesis of 3,3 '- [(3,4-dihydro-6-hydroxy-2,5,7,8-tetramethyl-2H-1-benzopyran-2-yl) methoxy-methyl] -benzene 2

To a solution Ha (3 g; 8.2 mmol) and of α, α'-dibromo-3-xylene (2.59 g, 9.85 mmol, 1.2 eq.) In a mixture of THF (~ 25 ml) and DMF (~ 25 ml) and maintained at -20 ° C is added NaH (360 mg; 9 mmol; 1 J eq.) under positive nitrogen pressure. The resulting solution is then stirred for 1 hour at -20 ° C and 5 hours at 0 ° C. At this time, the medium is diluted in ethyl acetate and hydrolyzed with water. The organic phase is then dried over MgSO4, filtered and evaporated. The oil obtained is purified by "flash chromatography" on a silica column (80/20 Petroleum Ether / Dichloromethane then 60/40 Petroleum Ether / Dichloromethane then 40/60 Petroleum Ether / Dichloromethane) to yield the majority fraction. product 2 - [(3-bromomethyl-benzyloxy) - methyl-3,4-dihydro-6 [(2-trimethylsilyl-ethoxy) -methoxy] -2, 5, 7, 8-tetramethyl-2H-1- benzopyran expected Via (P≈ Me3SiCH2CH2θCH2) (2.2 g; 50%). NMR (CDCl ₃ , 200 MHz, ppm): 7.32-7.23 (m, 4H); 4.96 (s, 2H); 4.57 (A ₂ B ₂ , 2H); 4.46 (s, 2H); 3.85 (m, 2H); 3.48 (A ₂ B ₂ , 2H); 2.59 (t, 6.9 Hz, 2H); 2J5 (s, 3H); 2.08 (s, 3H); 2.02 (s, 3H); 2.00-1.60 (m, 2H); 1.30 (m, 3H) .; 1.03-0.92 (m, 2H); 0.02 (s, 2H) Elementary Analysis for C28H4iBrθ4Si Calculated: C: 61.19; H: 7.52; Br: 14.53 Found: C: 61.06; H: 7.56; Br: 14.58

We obtain in the minority fraction the expected compound (1.08g 16%). If the stoichiometry of the reaction is 1 mole of alcohol per 0.5 of bromine derivative then the dimer is obtained almost exclusively (80%). The deprotection of the trimethylsilyl-ethoxy-methyl group to lead to 2 (88%) is carried out according to a procedure identical to that used in the last step of the synthesis of 1. NMR (CDCl ₃ , 200 MHz, ppm): 7.32-7.23 (m, 4H); 4.60 (AB ₂ , 4H); 4.18 (s, 2H); 3.48 (A ₂ B ₂ , 4H); 2.59 (t, 6.9 Hz, 4H); 2J5 (s, 6H); 2.08 (s, 12H); 2.02-1.92 (m, 2H); 1.85-1.60 (m, 2H); 1.30 (m, 6H). Elementary Analysis for Calculated C3gH5θOs: C: 75.23; H: 8.07 Found: C: 75.24; H: 8J 1

EXAMPLE 3

Synthesis of N, N'-diethyl-3 - [(3,4-dihydro-6-hydroxy-2,5,7,8-tetramethyl-2H-1-benzopyran-2-yl) -methoxy-methyl hydrochloride ] -benzylamine 3

To a solution of the brominated compound Via (400 mg; 0.73 mmol) and potassium carbonate (300 mg; 2.2 mmol, 3 eq.) In DMF (10 ml) is added at 0 ° C. diethylamine (98 μl; 0.95 mmol; 1.3 eq.). The resulting solution is stirred at room temperature for 16 hours then diluted in ether and washed with saturated NaCl solution until neutral pH. The organic phase is then dried over MgSO4, filtered and evaporated. The oil obtained is purified by "flash chromatography" on a silica column (95 / 4.5 / 0.5 Dichloromethane / MeOH / NH4θH) to provide in the majority fraction the compound N, N'-diethyl-3 - [(3,4- dihydro-6 - [(2-trimethylsilyl-ethoxy) -methoxy] -

2,5,7,8-tetramethyl-2H-1-benzopyran-2-yl) -methoxy-methyl-benzylamine (364 mg; 92%). Elementary Analysis for C32H5iNθ4Si Calculated: C: 70.93; H: 9.49; N: 2.59 Found C: 70.15; H: 9.43; N: 2.59

The deprotection of the trimethylsilyl-ethoxy-methyl group to lead to 3 (67%) is carried out according to a procedure identical to that used in the last stage of the synthesis of 1. The compound obtained in the form of the free base is hydrochloride with using a saturated solution of hydrochloric acid in ether. Melting point: 113 ° C IR (KBr, cm-): 3250, 2644, 2588, 1480, 1187, 796 NMR (CDCI3, 200 MHz, ppm): 12.27 (br m, 1H); 7.70-7.35 (m, 4H); 4.70-4.50

(m, 3H); 4.06 (s, 2H); 3.48 (A ₂ B ₂ , 2H); 3.15-2.75 (m, 4H); 2.59 (t, 6.9 Hz,

2H); 2J5 (s, 3H); 2J0 (s, 6H); 2.08-1.92 (m, 1H); 1.85-1.68 (m, 1H); 1.39

(t, 7.2 Hz, 6H); 1.30 (m, 3H)

Elementary Analysis for C26H38CINO3

Calculated: C: 69.70; H: 8.55; N: 3.13; Cl: 7.91 Found C: 68.45; H:

8.59; N: 3.06; Cl: 7.83

The examples from 4 to 9 which follow were carried out according to the same reaction sequence as for compound 3 by replacing diethylamine successively with methyl-aniline, (2-methoxy-phenyl) -piperazine, morpholine, l, 4-dioxa-8-azaspiro-4,5-decane, (E) -N-ethyl-N- (6,6-dimethyl-2-hepten-4-ynyl) -amine and l-oxa-2- oxo-3,8-diazaspiro- [4,5] -decane.

EXAMPLE 4 Synthesis of N-methyl-N-phenyl-3 - [(3,4-dihydro-6-hydroxy-2,5,7,8-tetramethyl-2H-1-benzopyran-2-yl) hydrochloride - methoxy-methyl] -benzylamine 4

Yield (1st stage 83% / 2nd stage 63%)

Melting point: 129 ° C

IR (KBr; cm ^"1 ): 3470, 2928, 1599, 1263, 1116

NMR (CDCI3, 200MHz, ppm): 7.45-7.05 (m, 9H); 4.60-4.30 (m, 5H); 3.48

(A ₂ B ₂ , 2H); 3.08 (s, 2H); 2.59 (t, 6.9 Hz, 2H); 2J5 (s, 3H); 2.10 (s, 6H); 2.00-1.65 (m, 2H); 1.30 (m, 3H)

Elementary Analysis for C29H36CINO3

Calculated: C: 72.26; H: 7.53; N: 2.90; Cl: 7.35 Found C: 70.73; H:

7.53; N: 2.77; Cl: 7.12 EXAMPLE 5

Synthesis of l - {[(3,4-dihydro-6-hydroxy-2,5,7,8-tetramethyl-2H-1-benzopyran-2-yl) - (3-benzy! Oxy-methyl)} - 4- (2-methoxy-phenyl) -piperazine 5

Yield (1st stage 67% / 2nd stage 70%) IR (KBr; cm "!): 3420, 2928, 2824, 1500, 1242, 1159, 748 NMR (CDCl ₃ , 200MHz, ppm): 7.40-7.20 (m, 4H); 7.05-6.80 (m, 4H); 4.60 (A ₂ B ₂ , 2H); 3.90 (s, 3H); 3.65-3.40 (m, 4H); 3.09 (brs, 4H); 2.75-2.55 (m , 6H); 2J5 (s, 3H); 2J0 (s, 6H); 2.00-1.65 (m, 2H); 1.30 (m, 3H) Elemental Analysis for C33H42N2O4

Calculated: C: 74.69; H: 7.98; N: 5.28 Found C: 74.03; H: 8.07; N: 5.09

EXAMPLE 6

Synthesis of l - [(3,4-dihydro-6-hydroxy-2,5,7,8-tetramethyl-) hydrochloride

2H-1-benzopyran-2-yl) -methoxy-methyl] -morpholine 6

Yield (1st stage 70% / 2nd stage 83%) Melting point: 168 ° C IR (KBr; cπr ¹ ): 3491, 2934, 2858, 2459, 1460, 1095

NMR (CDCI3, 200MHz, ppm): 13.15 (m, 1H); 7.60-7.35 (m, 4H); 4.64 (A ₂ B ₂ , 2H); 4.40-3.80 (m, 6H); 3.52 (A ₂ B ₂ , 2H); 3.25-3.05 (m, 2H); 2.85-2.55 (m, 4H); 2J5 (s, 3H); 2.10 (s, 6H); 2.00-1.65 (m, 2H); 1.30 (s, 3H) Elementary Analysis for C26H36CINO4

Calculated: C: 67.59; H: 7.85; N: 3.03; Cl: 7.67 Found C: 67.13; H

7.94; N: 3.02; Cl: 7.61

EXAMPLE 7

Synthesis of hydrochloride of α, α '- [(3,4-dihydro-6-hydroxy-2,5,7,8- tetramethyl-2H-1-benzopyran-2-yl) -methoxy] - (1,4 dioxa-8-azaspiro-4,5-decane) -xylene. 7

Yield (1st stage 82% / 2nd stage 70%)

Melting point: 107 ° C

IR (KBr; cm " ¹ ): 3383, 2930, 2542, 1452, 1257, 1116, 937

NMR (CDC13, 200MHz, ppm): 7.60-7.27 (m, 4H); 4.68 (brs, 1H); 4.60 (A ₂ B ₂ , 2H); 4.00-3.86 (m, 6H); 3.48 (A ₂ B ₂ , 2H); 3.35-3.15 (m, 2H); 2.95-2.80 (m,

2H); 2.65-2.30 (m, 6H); 2J5 (s, 3H); 2J0 (s, 6H); 2.00-1.65 (m, 2H); 1.30

(s, 3H)

Elementary Analysis for C9H40CINO5JH2O

Calculated: C: 64.97; H: 7.90; N: 2.61; Cl: 6.61 Found C: 64.81; H: 7.70; N: 2.56; Cl: 6.52

EXAMPLE 8

Synthesis of [(E) - N-ethyl-N- (6,6-dimethyl-2-hepten-4-ynyl] -3 [(3,4-dihydro-6-hydroxy-2,5,7,8 -tetramethyl-2H-1-benzapyran-2-yl) -methoxy-methyl] - benzylamine 8

Le spectre RMNiH du produit obtenu après purification sur colonne de silice est conforme à la structure du composé attendu.

The 1 H NMR spectrum of the product obtained after purification on a silica column conforms to the structure of the expected compound.

Elementary Analysis for C33H45NO3

Calculated: C: 78.68; H: 9.00; N: 2.78 Found C: 79.13; H: 9.04;

N: 2.69

EXAMPLE 9

Synthesis of α, α '- [(3,4-dihydro-6 - [(2-trimethylsiIyl-ethoxy) -methoxy] - (l-oxa-2-oxo-3,8-diazaspiro- [4,5] - decane) -xylene 9

The H NMR spectrum of the product obtained after purification on a silica column conforms to the structure of the expected compound.

EXAMPLE 10

Synthesis of 2- {3 - [(4-methyl-3-pentyn-1-oxy) -methyl] - (benzyloxy) -methyI} -3,4- dihydro-2,5,7,8-tetramethyI-2H- l-benzopyran-6-ol 10

To a suspension of NaH (41 mg; 1 mmol; 1.4 eq.) In 3 ml of THF is added at 0 ° C a solution of 4-methyl-3-pentene-1-ol (103 μl; 0.88 mmol; 1.2 eq .) in 5 ml of THF. The reaction is continued for 30 minutes at ambient temperature and then again cooled to 0 ° C. to add the bromine derivative Via (400 mg; 0.73 mmol) in solution in 2 ml of THF. The resulting medium is then stirred for 20 hours at room temperature then diluted in ether and washed with water to neutral pH. The organic phases were then dried over MgS04, ^'filtered and evaporated. The oil obtained is purified by "flash chromatography" on a silica column (85/15 Petroleum Ether / Ethyl acetate) to yield the product derived from 10 having a trimethylsilyl-ethoxy-methyl group (146 mg;

35%). Its deprotection is carried out according to a procedure identical to the synthesis of 1 (last step) and leads to 10 with a yield of 90%.

IR (KBr; cπr ¹ ): 3450, 2926, 2542, 1450, 1255, 1109

NMR (CDCl ₃ , 200MHz, ppm): 7.32-7.20 (m, 4H); 5.12 (t, 7.0 Hz, 1 H); 4.60

(A ₂ B ₂ , 2H); 4.49 (s, 2H); 4J9 (s, 1H); 3.53-3.39 (m, 4H); 2.60 (t, 6.8Hz,

2H); 2.35-2.25 (m, 2H); 2.15 (s, 3H); 2J0 (s, 6H) ^' ; 2.00-1.65 (m, 2H); 1.68

(s, 3H); 1.60 (s, 3H); 1.30 (s, 3H).

Elementary Analysis for C28H38O4

Calculated: C: 76.68; H: 8.73 Found C: 75.89; H: 9.04

EXAMPLE 11

Synthesis of 2- [2- (benzyloxy) -ethoxy-methyl] -3,4-dihydro-2,5,7,8-tetramethyl- 2H-1-benzopyran-6-ol 11

To a suspension of NaH (60% in oil; 520 mg; 13 mmol; 1.2 eq.) In DMF (40 ml) at - 30 ° C is added alcohol lia (4 g; 10.9 mmol) in solution in the DMF. The medium is stirred for 1 hour at -30 ° C. then the tert-butyl bromo-acetate (1.94 ml; 12 mmol; 1 J eq.) Is added slowly at this same temperature. The resulting solution is stirred for 24 hours at room temperature then the medium is diluted in ether and washed until neutral pH. The organic phases are then dried over MgSθ4, filtered and evaporated. The oil obtained is purified by "flash chromatography" on a silica column (90/10 Petroleum Ether / Ethyl Ether then 75/25 Petroleum Ether / Ethyl Ether) to provide the expected ester (3.26 g; 62%). NMR (CDCI3, 200MHz, ppm): 4.85 (s, 2H); 4.12 (A ₂ B ₂ , 2H); 3.90-3.75 (m, 2H); 3.60 (A ₂ B ₂ , 2H); 2.58 (t, 6.8 Hz, 2H); 2.21 (s, 3H); 2J5 (s, 3H); 2.10 (s, 3H); 2.00-1.65 (m, 2H); 1.30 (s, 3H); 1.02-0.93 (m, 2H); 0.02 (s, 9H) To a solution of the ester obtained above (2.6 g; 5.4 mmol) in 50 ml of THF is added at 0 ° C lithium aluminum hydride (1 M in THF ; 6 ml; 6 mmol; 1.1 eq). The reaction is stirred for 45 minutes at 0 ° C. and then hydrolyzed with wet Na2Sθ4. The volatiles are then evaporated and the oil obtained is purified on a silica column (80/20 Petroleum Ether / Ethyl Ether) to yield, in the majority fraction, the compound [(3,4-dihydro-6 - [(2-trimethylsilyl -ethoxy) - methoxy -2.5, 7,8-tetramethyl-2H-1-benzopyran-2-yl) -hydroxymethyl ethanol (1.5; 68%).

Elementary Analysis for C22H38θ5Si Calculated: C: 64.35; H: 9.33 Found C: 63.92; H: 9.24 Compound 11 is prepared by first acting on benzyl bromide on the alcohol obtained above and then deprotecting the trimethylsilyl-ethoxy-methyl group with sulfuric acid using procedures identical to the last two steps of the synthesis of 1. Yield (1st step 57% / 2nd step 81%) IR (film; cm " ¹ ): 3464, 2928, 1456, 1259, 1113, 736 NMR (CDC13, 200MHz, ppm): 7.30-7.20 (m, 5H); 4.56 (s, 2H); 4J9 (s, IH); 3.75-3.40 (m, 8H); 2.60 (t, 6.8 Hz, 2H); 2.15 (s, 3H); 2.10 (s, 6H); 2.05-1.65 (m, 2H); 1.30 (s, 3H) Elemental Analysis for C23H30O4 Calculated: C: 74.56; H: 8.16; Found C: 73.95; H: 8.10 The following examples 12 and 13 were carried out according to the same reaction sequence as for compound 11, replacing the benzyl bromide by geranyl bromide or farnesyl bromide.

EXAMPLE 12 Synthesis of 2- [2- (3,7-dimethyl-octa-2,6-dienyloxy) -ethoxy-methyl] -3,4-dihydro- 2,5,7,8-tetramethyl-2H-1- benzopyran-6-ol 12

Yield (1st stage 75% / 2nd stage 83%)

IR (film; cm " ¹ ): 3450, 2930, 1480, 1113

NMR (CDC13, 200MHz, ppm): 5.42-5.32 (m, 1H); 5.17-5.05 (m, 1H); 4.24 (s,

IH); 4.05 (d, 6.4 Hz, ppm); 3.78-3.68 (m, 2H); 3.62-3.44 (m, 4H); 2.65 (t, 6.8

Hz, 2H); 2J5 (s, 3H); 2J0 (s, 6H); 2.05-1.58 (m, 13H); 1.30 (s, 3H)

Elementary Analysis for C22H39O4

Calculated: C: 75.14; H: 9.46; Found C: 74.28; H: 9.67

EXAMPLE 13

Synthesis of 2- [2- (3,7, ll-trimethyl-dodeca-2,6,10-trienyloxy-ethoxymethyl] -3,4- dihydro-2,5,7,8 tetramethyl-2H-1-benzopyran- 6-ol 13

Yield (1st step 50% / 2nd step 90%) IR (film; cm " ¹ ): 3439, 2924, 1450, 1261, 1087

NMR (CDC13, 200MHz, ppm): 5.42-5.32 (m, 1H); 5.17-5.05 (m, 2H); 4.20 (s, 1H); 4.03 (d, 6.4 Hz, 2H); 3.75-3.35 (m, 6H); 2.60 (t, 6.8 Hz, 2H); 2.20-1.50 (M, 31H); 1.30 (s, 3H) Elementary Analysis for C31H48O4 Calculated: C: 76.82; H: 9.98; Found C: 77.05; H: 10.07 EXAMPLE 14

Synthesis of 2- [2- (3,7-dimethyl-octa-2,6-dienyl-amino) -ethoxymethyI] -3,4- dihydro-2,5,7,8-tetramethyI-2H-1-benzopyran- 6-ol 14

To a solution of alcohol 1 (lg; 1.2 eq.) In THF (5 ml) at 0 ° C is added triethylamine (1.4 ml; 10 mmol, 5 eq.) Then mesyl chloride (0.22 ml; 1.4 eq.). After 1 h of stirring at 0 ° C., the medium is brought to room temperature and filtered through celite, washing with ethyl ether. Before evaporation,

5 ml of DMF are added to the filtrate, then the THF and the ether are eliminated in the cold on a rotary evaporator.

The remaining solution is then added to the sodium anion of N-trifluoroacetyl-geranylamine generated by the action of sodium hydride (60% dispersion in oil; 96 mg) on N-trifluoroacetyl-geranylamine (500 mg; 2 mmol) in DMF (5 ml) at 0 ° C. The resulting medium is heated at 60 ° C for 12 h then diluted in ether and extracted with water until neutral pH. The organic phases are then dried over MgSO4, filtered and evaporated. The oil obtained is purified by "flash chromatography" on a silica column (50/50 Petroleum Ether / Ethyl acetate) provides the amide with a yield of 55%.

The deprotection of the trifluoroacetyl group is carried out by the action of KOH (46 mg;

6 eq) on a solution of amide (280 mg; 0.44 mmol) in methanol (4 ml) at room temperature for 1 h. After evaporation of the methanol, dilution in ether and washing with water until pH - 8-9. The organic phases are then dried over MgSθ4, filtered and evaporated. The oil obtained is purified by "flash chromatography" on a silica column (95 / 4.5 / 0.5 CH2Cl2 / MeOH / NH4θH) to provide the expected amine (84%). The deprotection of the trimethylsilyl-ethoxy-methyl group is carried out according to the standard conditions described in the synthesis of 1 to lead after purification (95 / 4.5 / 0.5 CH ₂ Cl2 / MeOH / NH ₄ OH) to 14 (92%). IR (film; cm-1): 3420, 2922, 1452, 1257, 1089 NMR (CDCI3, 200MHz, ppm): 5.42-5.32 (m, 1H); 5J7-5.05 (M, 1H); 3.65-

3.55 (m, 2H); 3.45 (A ₂ B ₂ , 2H); 3.20 (d, 6.4 Hz, 2H); 2.75 (t, 5.0 Hz, 2H);

2.60 (t, 6.8 Hz, 2H); 2.20-1.50 (M, 22H); 1.30 (s, 3H)

Elementary Analysis for C26H41NO3

Calculated: C: 75.14; H: 9.94; N: 3.37 Found C: 74.31; H: 9.92; NOT :

3.63

EXAMPLE 15

Synthesis of 2- [2- (N-ethyl-3,7-dimethyl-octa-2,6-dienyl-amino) -ethoxy-methyl] -

3,4-dihydro-2,5,7,8-tetramethyl-2H-1-benzopyran-6-ol 15

NOTE: The synthesis of this compound requires the use of N-acetylgeranylamine which is prepared by the action of acetic anhydride on geranyamine.

The synthesis of this compound is carried out according to the same reaction sequence as for

14. Only the step of deprotection of the trifluoroacetyl group is replaced by the reduction of the acetyl group with lithium aluminum hydride in THF for 1 h at room temperature.

Yield (1st stage 55% / 2nd stage 60% / 3rd stage 85%)

IR (film; cm-1): 3420, 2922, 1452, 1257, 1089

NMR (CDC13, 200MHz, ppm): 5.42-5.32 (m, 1H); 5J7-5.05 (M, 1H); 3.65-

3.55 (m, 2H); 3.45 (A ₂ B ₂ , 2H); 3.20 (d, 6.4 Hz, 2H); 2.70-2.50 (m, 6H); 1.30 (s, 3H); 1.03 (t, 7.4 Hz, 3H)

Elementary Analysis for C28H45NO3

Calculated: C: 75.80; H: 10.22; N: 3.16 Found C: 75.25; H: 10.23;

N.3.20 EXAMPLE 16

Synthesis of 2 - [(E) -N-ethyl-N- (6,6-dimethyl-2-hepten-4-ynyl) -amino-ethoxymethyl] -3,4-dihydro-2,5,7,8- tetramethyl-2H-1-benzopyran-6-ol 16

The synthesis of derivative 16 is carried out according to the same reaction sequence (step # 1 and

# 3) only for 14.

However, in the first step, the amine is reacted with a solution of the preformed mesylate in DMF in the presence of K1 (1 eq.) And potassium carbonate (4 eq.) Then the resulting medium is heated to 100 ° C for 8 h.

Yield (1st stage 32% / 2nd stage 60%)

IR (film; cm-1): 3420, 2972, 1468, 1381, 1095

NMR (CDC13, 200MHz, ppm): 6J2 (dt, 15.8 Hz / 6.5 Hz, 1H); 5.67 (d, 15.8 Hz, 1 H); 3.70-3.35 (m, 4H); 3J2 (d, 6.5 Hz, 2H); 2.68-2.45 (m, 6H); 2J5 (s, 3H)

; 2J0 (s, 6H); 2.06-1.95 (m, 1H); 1.87-1.73 (m, 1H); 1.30 (s, 3H); 1.25 (s,

9H); 1.05 (t, 7Hz, 3H)

Elementary Analysis for C27H41NO3

Calculated: C: 75.83; H: 9.66; N: 3.28 Found C: 75.20; H: 9.64; N: 3.04

EXAMPLE 17

Synthesis of {[(3,4-dihydro-6-hydroxy-2,5,7,8-tetramethyl-2H-1-benzopyran-2-yl) -methoxy methyljcarbonyl} -

[(N, N-dimethyl-amino) -ethyl] -amine 17

17 Une solution d'ester (4,4 g ; 9J2 mmoles) dans le dichlorométhane (45 ml) en présence d'acide trifluoroacétique (14 ml ; 182 mmoles ; 20 éq.) est agitée pendant

17 A solution of ester (4.4 g; 9J2 mmol) in dichloromethane (45 ml) in the presence of trifluoroacetic acid (14 ml; 182 mmol; 20 eq.) Is stirred for

6 h at room temperature. The trifluoroacetic acid is then co-evaporated with 3 x 300 ml of toluene and the oil obtained purified on a silica column (50/50

Petroleum Ether / Ethyl acetate then 100 Ethyl acetate) to yield 2 - [(3,4-dihydro-6-hydroxy-2,5,7,8-tetramethyl-2H-1-benzopyran acid) -2-yl) - methoxy] acetic Xla (n = 1) (75%).

Elementary Analysis for C16H22NO5

Calculated: C: 65.29; H: 7.53 Found C: 64.42; H: 7.55

1J'-carbonyldiimidazole (303 mg; 1.87 mmol; 1 J eq.) Is added to a solution of Xla acid previously synthesized (500 mg; 1.7 mmol) and THF

(10 ml). After stirring for 1 hour at room temperature, an amine solution

(185 μl; 1.70 mmol; 1 eq.) In THF (8 ml) is added slowly to the medium.

The resulting solution is stirred for 24 h at room temperature then diluted in ether and washed with a saturated solution of sodium chloride.

The organic phases are then dried over MgSθ4, filtered and evaporated.

The oil obtained is purified by "flash chromatography" on a silica column

(85/15 CH2θ2 / MeOH) to provide a compound (422 mg); whose spectrum

NMR H corresponds to the structure of product 17.

Examples 18 and 19 which follow were carried out according to the same procedure as for the preparation of compound 17, replacing N, N'-dimethyl-amino-ethyl-amine with phenethylamine or 3-aminomethyl-pyridine (3- picolylamine).

EXAMPLE 18

Synthesis of {[(3,4-dihydro-6-hydroxy-2,5,7,8-tetramethyl-2H-1-benzopyran-2-yl) -methoxy-methyl] -carbonyl} -phenethyl-amine 18

18 Rendement (84%)

18 Yield (84%)

IR (film, cm ' ¹ ): 3395, 2930, 1674, 1541, 1454, 1257, 1089 NMR (CDC13. 200MHz, ppm): 7.35-7.10 (m, 5H); 4.37 (OJH); 4.01 (A2B2, 2H); 3.65-3.40 (m, 4H); 2.79 (t, 7.1 Hz, 2H); 2.60 (t, 6.8 Hz, 2H); 2J6 (s, 3H); 2.12 (s, 3H); 2J0-2.05 (m, 5H); 1.95-1.60 (m, 2H); 1.20 (s, 3H) Elemental Analysis for C24H31NO4-I / 3C4H8O2

Calculated: C: 71.28; H: 7.95; N: 3.28 Found C: 70.98; H: 7.87; N: 3.42

EXAMPLE 19

Synthesis of {[(3,4-dihydro-6-hydroxy-2,5,7,8-tetramethyl-2H-1-benzopyran-2-yl) -methoxy-methyl jcarbonyl} -3-pyridyl-hydrochloride -amine 19

Melting point: 100 ° C

Yield (74%)

IR (KBr, cm ' ¹ ): 3391, 2928, 2361, 1665, 1541, 1118

NMR (dβ-DMSO, 200MHz. Ppm): 8.79 (brs, 2H); 8.45-8.25 (m, 2H); 8.0-7.85

(m, 1H); 4.48 (d, 6 Hz, 2H); 4.07 (A ₂ B ₂ , 2H); 3.60 (A ₂ B ₂ , 2H); 2.51 (m,

2H); 2.04 (s, 3H); 2.02 (s, 3H); 2.0-1.60 (m, 5H); 1.21 (s, 3H)

Elementary Analysis for C22H29CIN2O4-I.3H2O

Calculated: C: 59.47; H: 7.17; N: 6.30; Cl: 7.98 Found C: 59.18; H:

7.19; N: 6.39; Cl: 8.19 EXAMPLE 20

Synthesis of 2- [2- (3-pyridyl-methyl-amino) -ethoxy-methyl] hydrochloride -

(3,4-dihydro-2,5,7,8-tetramethyl-2H-1-benzopyran-6-ol 20

Compound 20 (226 mg; 0.58 mmol) is prepared by reduction of 19 with lithium aluminum hydride (1 M in THF; 1.2 ml; 1.2 mmol; 2 eq) for 2 h at room temperature. After hydrolysis with wet Na2SO4 and evaporation, the oil collected and purified on a silica column (95 / 4.5 / 0.5

CH2Cl2 MeOH / NH4θH) to yield the expected compound (47%).

NMR (CDCI3. 200MHz. Ppm): 8.58 (brs, 1H); 8.36 (brs, 1H); 8.04 (m, 1H);

7.60 (brs, 1 H); 4.60-4.52 lm. _. 2H); 4.32-4.20 (m, 2H); 4.05 (A ₂ B ₂ , 2H); 3.55

(A ₂ B ₂ , 2H); 2.60 (t, 6.8Hz, 2H) 2J0 (s, 3H); 2.07 (s, 3H); 2.04 (s, 3H); 2.01-1.65 (m, 2H); 1.30 (s, 3H)

Elementary Analysis for C22H32N2O3-I .7HCI

Calculated: C: 61.08; H: 7.45; N: 6.47 Found C: 61.70; H: 7.31; NOT :

6.40

EXAMPLE 21

Synthesis of [l-oxa-2-oxo-3,8-diazaspiro [4,5] -decane-8-yl] -ethoxymethyl] -3,4- dihydro-2,5,7,8-tetramethyl-2H- l-benzopyran-6-ol 21

This compound is prepared from the acid Xla according to the same sequence of steps as for the preparation of compound 20 by first reacting Xla with l-oxa-2-oxo-3,8-diazaspiro [4,5] -decane then by reducing the intermediate formed by LiAlH4.

The NMR * H spectrum of the product obtained after purification on a silica column conforms to the structure of the expected compound.

BIOLOGICAL ACTIVITIES

Pharmacological results

The antioxidant activity of the derivatives of the present invention has been more particularly demonstrated on microsomes of rat livers, following a chemical peroxidation induced by ferric ions and on human LDL. The inhibitory action of the compounds of the present invention with respect to the oxidation of human LDL has been demonstrated whether it is following a chemical oxidation by copper sulphate or following a biological oxidation by human endothelial cells from umbilical veins.

1. Inhibition of the oxidation of microsomes of rat liver by ferric lesions The study was carried out according to the technique described in the reference "Antioxidative properties of harmane and carboline alkaloids"; S. Y. H. Tse, I-T. Mak, B-J. Dickens, Biochem Pharmacol. 42 (3), 459-464, 1991. The activity of the compounds was tested in comparison with vitamin E. Many compounds of the present invention showed an activity superior to vitamin E in this test as indicated some examples in the table below:

COMPOUNDS IC ₅₀ (μM)

Vitamin E 2.3

Example 10 0.3

Example 13 0J

Example 16 0.03

Example 12 0.2

Example 4 0.06 2. Inhibition of oxidation of human LDL by copper sulphate The studies were carried out with human LDL incubated with copper sulphate (10 μM) for 6 hours at 37 ° C. with or without test compound. Lipid peroxidation is evaluated by the TBARS technique as described by C. Breugnot et al., Biochem. Pharmacol. 4fl, 1975-1980 (1990). The antioxidant activity of the compounds of the present invention has been studied in comparison with vitamin E. The few illustrative examples given in the table below show that the derivatives of the present invention have a favorable antioxidant profile vis- with regard to vitamin E:

COMPOUNDS IC ₅₀ (μM)

Vitamin E 7.4

Example 13 3.00

Example 16 1J8

Example 8 0.84

Example 12 0.3

Example 15 0.3

3. Inhibition of oxidation of human LDL by endothelial cells of human umbilical vein in culture Human LDL are stored at 4 ° C. in a buffer (150 mM NaCl, 0.01% EDTA, pH = 7.4) at 4 °. vs. Before use, one volume of LDL is dialyzed for 24 hours at 4 ° C. against 5 x 1000 volumes of buffer (NaCl 136.8 mM, KC1 2.68 mM, KH2PO4 1.47 mM, Na ₂ HPθ4 8.09 mM, pH = 7). LDL are used at the final protein concentrations (Coomassie, Pierce test) of 25 or 50 μg / ml.

The endothelial cells, line ECV304 (ATCC CRL-1998) are cultivated in wells of 35 mm in diameter in a DMEM-HEther of Petroleum ES- Glutamax I medium supplemented with a fetal calf serum 10%, penicillin- streptomycin 100 IU / ml - 100 μg / ml, and fungizone 2.5 μg / ml up to a sub-state meet. The cells are then incubated for 24 hours in DMEM-HEther of PetroleumES-Glutamax I medium containing ultroser G 2%; then, the oxidation of LDL is triggered by their contact with the cells for 24 hours in the nutrient medium HAM F-10 (Breugnot C, Maziere C, Salmon S., Auclair M., Santus R., Morliere P. , Lenaers A. and Mazière JC: Phenothiazines inhibit copper and endothelial cell-induced peroxidation of lo density lipoprotein. A comparative study with probucol, butylated hydroxy toluene and vitamin E. Biochemical Pharmacology, 1990, 40: 1975-1980). The culture supernatant is collected, centrifuged (2000 xg, 10 minutes, 4 ° C), and the reaction with thiobarbituric acid (0.12 N, in 10 mM TRIS buffer; pH = 7) is carried out on the supernatant after acidification (final pH = 2-3) with 15% trichloroacetic acid (Yagi K.: Lipid peroxides and human diseases. Chemistry and physics of lipids, 1987, 45: 337-351). The samples are heated for 30 minutes at 95 ° C., the TBARs are extracted with butanol-1 and quantified by fluorimetry (excitation wavelengths 515 nm, emission wavelength 548 nm, Perkin Elmer LS-50B spectrofluorimeter). The results are expressed in nmol malondialdehyde equivalent / mg LDL proteins.

The antioxidant activity of ^~ compounds of the present invention in this model was studied in comparison with vitamin E. As shown by some illustrative examples in the table below, the compounds of the present invention were very much superior to vitamin E in this test.

COMPOUNDS IC ₅₀ (μM)

Vitamin E 15

Example 16 0.03

Example 8 0.3

Example 12 0, 1

Example 15 0.05

Example 14 0.1 These results clearly indicate the greater potency of certain derivatives forming part of the present invention compared to vitamin E as lipid protectors such as microsomes or human LDL against oxidative modifications and in particular during biological oxidation mediated by human endothelial cells.

The oxidative changes in LDL today clearly appear to play an important role in the genesis and development of vascular lesions such as atheromas. Consequently, the antioxidant properties, in particular at the LDL level, of the derivatives of the present invention allow their use as medicaments in the treatment and / or prevention of atherosclerosis including its various peripheral vascular localizations, coronary or cerebral, diseases due vascular complications linked to lipoproteins, but also pathologies in which a membrane lipid peroxidation plays an initiating and / or aggravating role such as ischemical heart disease, organ reperfusion, including transplant, ischemical traumatic pathologies of the central nervous system or peripheral, autoimmune diseases and metabolic diseases such as diabetes.

Furthermore, the derivatives of the present invention have been shown to be particularly effective in neutralizing hydroxyl radicals (OH): this is how, in a test making it possible to determine the ability of compounds to trap the hydroxyl radical (method using the assay of the radical 2-hydroxy-5,5 dimethyl-1-pyrroline-N-oxide by RPE according to Miyagawa et al., J. Clin. Biochem. Nutr. 5, 1-7, 1988), certain products of the present invention are are shown to be more active than vitamin E or BHT; by way of example, the table below gives the comparative activities of these two reference products and of example 8 of the present invention.

COMPOUNDS CI ₃₀ (μM)

Vitamin E 65

BHT 46

Example 8 6.4 Hydroxyl radicals constitute a source of extremely aggressive radicals at the cellular and molecular levels and, therefore, the compounds of the present invention which have the advantage of being both inhibitors of lipid peroxidation and free radical scavengers are also found. their usefulness in dermatology and dermo-cosmetology and in the treatment of various pathologies such as certain forms of cancer (in particular of tumors linked to ionizing radiation such as certain skin cancers), the aging of tissues (in particular the aging of the skin), and certain forms of degeneration such as Parkinson's or Alzheimer's disease. As such, various antioxidants such as vitamin E have shown a protective effect in a model of MPTP-induced neurotoxicity (cf. TL Perry et al, Neurosc. Letters, 6fl, 109, 1985) model considered as predictive of neuro diseases - degenerative (cf. Neuroscience Facts, 2 (23), 89, 1992). The greater inhibitory activity of oxidative processes observed with many derivatives of the present invention compared to vitamin E therefore makes them particularly useful for the treatment and / or prevention of neurodegenerative diseases.

The present invention also relates to pharmaceutical compositions containing as active ingredient a compound of general formula I or one of its acceptable salts for pharmaceutical use, mixed or associated with a suitable excipient. These compositions can take, for example, the form of solid, liquid compositions, emulsions, lotions or creams. As solid compositions for oral administration, tablets, pills, powders (gelatin capsules, cachets) or granules can be used. In these compositions, the active principle according to the invention is mixed with one or more inert diluents, such as starch, cellulose, sucrose, lactose or silica, under a stream of argon. These compositions can also comprise substances other than diluents, for example one or more lubricants such as magnesium stearate or talc, a dye, a coating (dragees) or a varnish.

As liquid compositions for oral administration, there may be used pharmaceutically acceptable solutions, suspensions, emulsions, syrups and elixirs containing inert diluents such as water, ethanol, glycerol, vegetable oils or oil paraffin. These compositions can include substances other than diluents, for example wetting, sweetening, thickening, flavoring or stabilizing products. The sterile compositions for parenteral administration can preferably be aqueous or non-aqueous solutions, suspensions or emulsions. As solvent or vehicle, water, propylene glycol, a polyethylene glycol, vegetable oils, in particular olive oil, injectable organic esters, for example ethyl oleate or other suitable organic solvents, can be used. These compositions can also contain adjuvants, in particular wetting agents, isotonizers, emulsifiers, dispersants and stabilizers. Sterilization can be done in several ways, for example by aseptic filtration, by incorporating sterilizing agents into the composition, by irradiation or by heating. They can also be prepared in the form of sterile solid compositions which can be dissolved at the time of use in sterile water or any other sterile injectable medium. The compositions for rectal administration are suppositories or rectal capsules which contain, in addition to the active product, excipients such as cocoa butter, semi-synthetic glycerides or polyethylene glycols.

The compositions for topical administration can be, for example, creams, lotions, eye drops, mouthwashes, nasal drops or aerosols. The doses depend on the desired effect, on the duration of the treatment and on the route of administration used; they are generally between 0.001 g and 1 g (preferably between 0.005 g and 0.25 g) per day, preferably orally for an adult with unit doses ranging from 0J mg to 500 mg of active substance, preferably from 1 mg to 50 mg.

In general, the doctor will determine the appropriate dosage based on age, weight and all other factors specific to the subject to be treated. The following examples illustrate compositions according to the invention

[in these examples, the term "active component" denotes one or more

(generally one) of the compounds of formula (I) according to the present invention]:

Tablets

They can be prepared by direct compression or by passing through wet granulation. The direct compression procedure is preferred, but it may not be suitable in all cases depending on the doses and the physical properties of the active component. A - By direct compression mg for 1 tablet active component 10.0 microcrystalline cellulose PCB 89.5 magnesium stearate 0.5

100.0 The active component is passed through a sieve with a mesh opening of 250 μm side, it is mixed with the excipients and it is compressed using 6.0 mm punches. Tablets with other mechanical strengths can be prepared by varying the compression weight with the use of appropriate punches.

B - Wet granulation mg for one tablet active component 10.0 lactose Codex 74.5 starch Codex 10.0 corn starch pregelatinized Codex 5.0 magnesium stearate L5.

Compression weight 100.0 The active component is passed through a sieve with a mesh opening of 250 μm and mixed with lactose, starch and pregelatinized starch. The mixed powders are moistened with purified water, they are granulated, dried, sieved and mixed with magnesium stearate. The lubricated granules are put into tablets as for the formulas by direct compression. A coating film can be applied to the tablets using suitable film-forming materials, for example methylcellulose or hydroxypropyl-methyl-cellulose, according to conventional techniques. Sugar tablets can also be coated.

Mg capsules for one active component capsule 10.0

* starch 1500 89.5 magnesium stearate Codex £ 5 Filling weight 100.0 * a form of directly compressible starch from the firm Colorcon Ltd, Orpington, Kent, United Kingdom.

The active component is passed through a sieve with a mesh opening of 250 μm and mixed with the other substances. The mixture is introduced into hard gelatin capsules No. 2 on an appropriate filling machine. Other dosage units can be prepared by changing the filling weight and, if necessary, changing the size of the capsule.

Syrup mg per 5 ml dose active ingredient 10.0 sucrose Codex 2750.0 glycerin Codex 500.0 buffer) flavor) color) q.s. preservative) distilled water 5.0

The active component, the buffer, the flavor, the color and the preservative are dissolved in part of the water and the glycerin is added. The remainder of the water is heated to 80 ° C. and the sucrose is dissolved therein and then cooled. The two solutions are combined, the volume is adjusted and mixed. The syrup obtained is clarified by filtration.

Suppositories

Active ingredient 10.0 mg

* Witepsol H 15 supplement to 1.0 g

* Brand sold for Adeps Solidus of the European Pharmacopoeia.

A suspension of the active component in Witepsol H 15 is prepared and introduced into a suitable machine with 1 g suppository molds.

Liquid for administration by intravenous injection g / 1 active component 2.0 water for injection Codex supplement to 1000.0 Sodium chloride can be added to adjust the tone of the solution and adjust the pH to maximum stability and / or to facilitate the dissolution of the active component by means of a dilute acid or alkali or by adding buffer salts. appropriate. The solution is prepared, clarified and introduced into ampoules of appropriate size which are sealed by melting the glass. The liquid for injection can also be sterilized by heating in an autoclave according to one of the acceptable cycles. The solution can also be sterilized by filtration and introduced into a sterile ampoule under aseptic conditions. The solution can be introduced into the ampoules in a gaseous atmosphere.

Cartridges for inhalation g / cartridge active component micronized 1, 0 lactose Codex 39.0 The active component is micronized in a fluid energy mill and brought to the state of fine particles before mixing with lactose for tablets in a high blender energy. The powder mixture is introduced into hard gelatin capsules No. 3 on an appropriate encapsulating machine. The contents of the cartridges are administered using a powder inhaler.

mg / dose for 1 box of micronized active ingredient 0.500 120 mg oleic acid Codex 0.050 12 mg trichlorofluoromethane for pharmaceutical use 22.25 5.34 g

dichlorodifluoromethane for pharmaceutical use 60.90 14.62g The active component is micronized in a fluid energy mill and placed in the form of fine particles. The oleic acid is mixed with the trichlorofluoromethane at a temperature of 10-15 ° C. and the micronized drug is introduced into the solution using a mixer with a high shearing effect. The suspension is introduced in measured quantity into aluminum aerosol cans on which are fixed appropriate metering valves delivering a dose of 85 mg of the suspension; dichlorodifluoromethane is introduced into the boxes by injection through the valves.

Claims

1. Compounds corresponding to general formula I

in which n represents an integer between 1 and 10 R represents CH2OR1, CONR1R2, CH2NR1R2 or Ari ⁱⁿ which Ri, R2, which are identical or different, each represent a hydrogen, a linear or branched alkyl radical comprising from 1 to 20 carbon atoms which may itself contain one or more unsaturations such as doubles or triple bonds and can be substituted by an aromatic residue (such as a phenyl, a pyridine or a benzopyran which is variously substituted) by a halogen (chlorine, bromine or fluorine), by an alkoxy, (OR4) by a thiother (SR4) , with a silane (SiR4R5R6) or with an amine (NR4R5) in which R4, R5 and Rβ represent an alkyl or aryl radical and, RI and R2, when attached to the same nitrogen atom can also form a variously functionalized cycle such as a 3.8 diazo-l-oxa-2-oxaspiro [4,5] decane, Ari represents an aromatic radical such as a phenyl which can be variously substituted in various positions by a halogen (chlorine, bromine, fluorine or iodine), an amine (NR1R2), an alkoxy (OR), a thiother (SRi), a radical linear or branched alkyl comprising from 1 to 10 carbon atoms in a straight or branched chain which can itself be substituted by an alkoxy (ORi), an amine (NR1R2), or a thioether (SRi) and moreover, Ri and R2 can also form a cycle with the nitrogen to which they are attached, as well as their salts, hydrates, solvates and bioprecursors acceptable for therapeutic use, the compounds of formula (I) containing 1 or more centers asymmetrical, which may take the form of racemic mixtures or enantiomers. 2. A compound of formula (I) according to claim 1 selected from 2 - [(benzy loxy) -methyl] -3, 4-dihydro-2, 5, 7, 8-tetramethy 1-2H- 1 - benzopyran-6-ol 3, 3 '- [(3, 4-dihydro-6-hydroxy-2, 5.7, 8-tetramethyl-2H- 1 -benzopyran-2-yl) methoxy-methyl] -benzene N, N'-diethyl-3 - [(3,4-dihydro-6-hydroxy-2,5,7,8Jetramethyl-2H-1-benzopyran-2-yl) -methoxy-methyl] -benzylamine N-methyl 1 -N-pheny l-3 - [(3, 4-dihydro-6-hydroxy-2, 5, 7, 8-tetramethy 1-2H- 1 - benzopyran-2-yl) -methoxy-methyl] -benzylamine 1 - {[(3, 4-dihydro-6-hydroxy-2, 5, 7, 8-tetramethyl-2H- 1 -benzopyran-2-y 1) - (3-benzyloxy-methyl)} - 4- (2-methoxy-phenyl) -piperazine 1 - [(3, 4-dihydro-6-hydroxy-2, 5, 7, 8-tetramethy 1-2H- 1 -benzopyran-2-y 1) - methoxy-methylj-morpholine α, α '- [(3 , 4-dihydro-6-hydroxy-2,5,7,8-tetramethyl-2H-1-benzopyran-2-yl) - methoxy] - (1, 4-dioxa-8-azaspiro-4, 5-decane) -xylene [(E) - N-ethyl-N- (6,6-dimethyl-2-hepten-4-ynyl] -3 [(3,4-dihydro-6-hydroxy-2,5,7,8-tetramethyl- 2H-1-benzapyran-2-yl) -methoxy-methyl] - benzylamine α, α '- [(3, 4 -dihydro-6 - [(2-trimethylsilyl-ethoxy) -methoxy] - (1 -oxa-2 - oxo-3,8-diazaspiro [4,5] -decane) -xylene 2- {3 - [(4-methyl-3-pentyn-1-oxy) -methyl] - (benzyloxy) -methyl} -3,4- dihydro-2, 5, 7, 8-tetramethy 1-2H- 1 -benzopyran-6-ol 2- [2- (benzyloxy) -ethoxy-methyl] -3,4-dihydro-2,5,7,8-tetramethyl-2H-1- benzopyran-6-ol 2- [2- (3,7-dimethyl-octa-2,6-dienyloxy) -ethoxy-methyl] -3,4-dihydro- 2,5,7,8-tetramethy 1-2H- 1 -benzopyran-6-ol 2- [2- (3,7J l-trimethyl-dodeca-2,6J0-trienyloxy-ethoxymethyl] -3,4- dihydro-2,5,7,8 tetramethyl-2H-l-benzopyran-6-ol 2- [2- (3,7-dimethyl-octa-2,6-dienyl-amino) -ethoxymethyl] -3,4-dihydro- 2,5,7,8-tetramethy 1-2H- 1 -be nzopy ran-6-ol 2- [2- (N-ethyl-3,7-dimethyl-octa-2,6-dienyl-amino) -ethoxy-methyl] -3,4- dihydro-2, 5, 7, 8-tetramethyl-2H- 1 -benzopyran-6-ol 2 - [(E) -N-ethyl-N- (6,6-dimethyl-2-hepten-4-ynyl) -amino-ethoxymethyl] -3,4- dihydro-2,5 , 7,8-tetramethyl-2H-1-benzopyran-6-ol {[((3, 4-dihydro-6-hydroxy-2, 5, 7, 8-tetramethyl-2H- 1 -benzopy ran-2-yl) - methoxy methyl] carbonyl} - [(N, N-dimethyl-amino ) -ethyl] -amine {[(3, 4-dihydro-6-hydroxy-2, 5, 7, 8-tetramethyl-2H- 1 -benzopyran-2-y 1) - methoxy-methylj-carbonyl} -phenethy 1 -amine {[(3,4-dihydro-6-hydroxy-2,5,7,8-tetramethyl-2H-1-benzopyran-2-yl) - methoxy-methyl 1] carbonyl} -3-pyridyl-methyl 1 -amine 2- [2- (3-pyridyl-methyl-amino) -ethoxy-methyl] - (3,4-dihydro-2,5,7,8- tetramethyl-2H- 1 -benzopy ran-6-ol [l-oxa-2-oxo-3,8-diazaspiro [4,5] -decane-8-yl] -ethoxymethyl] -3,4-dihydro- 2,5,7,8-tetramethyl-2H- 1 - benzopyran-6-ol 3. Compounds according to claim 1 characterized in that n represents 1. 4. Compounds according to claim 1 characterized in that R represents CH ₂ OR !. 5. Compounds according to claim 1, characterized in that R represents CH ₂ NRιR ₂ or CONRιR ₂ . 6. Compounds according to claim 1 characterized in that R represents Ar ^ 7. Compounds according to claim 6 characterized in that Ari represents a phenyl substituted by a methylamine (CH2NR1R2). 8. Compounds according to claim 6, characterized in that Ari represents a phenyl substituted with a methyl ether (CH2OR1). 9. A method of manufacturing the compounds of formula I according to claims 1 to 8 characterized in that one transforms an intermediate of formula I '

in which n and R are defined as above and P represents a protective group in compound of formula (I) by the methods and techniques appropriate for this type of transformation depending on the nature of P. 10. A method of manufacturing the compounds of formula (I ') according to claim 9 wherein R represents Ari characterized in that an alcohol of formula (II) is condensed

with an electrophile of formula Y (CH2) _n rι in which Y represents a leaving group such as a halogen (bromine, chlorine or iodine), a tosylate, a mesylate or a triflate in the presence of an organic or inorganic base. 11. Method for manufacturing the compounds of formula (T) according to claim 9 but in which R represents CH2OR1 characterized in that an alcohol of formula (X) is condensed

with an electrophile of formula Y - Ri in which n, Ri and Y are defined as above in the presence of an organic or inorganic base. 12. Process for the preparation of the compound of formula (I) according to claim 1 but in which R represents C (O) NRιR2 characterized in that an acid of formula (XI) or an activated ester derived from this acid is condensed

with an amine HNR1R2 according to the methods and techniques appropriate for the preparation of an amide from an acid. 13. A method of preparing the compounds of formula (I ') according to claim 1 but in which R represents CH2NR1R2 characterized in that an amine of formula R1R2NH is condensed with an electrophile of formula (XII) in which Y remains a group therefore, in the presence of an organic or inorganic base. 14. Process for the preparation of the compounds of formulas I characterized in that a compound of general formula I or I ′ or a salt or a derivative comprising a protecting group of such a compound is converted into another compound of general formula (I). 15. Pharmaceutical compositions containing as active ingredient a derivative according to claims 1 to 8 with suitable pharmaceutical excipients. 16. Pharmaceutical compositions containing, as active ingredient, a derivative of formula (I) according to claims 1 to 8, in combination with a pharmaceutical carrier acceptable for the treatment and / or prevention of dyslipidemia and atherosclerosis. 17. Pharmaceutical compositions containing, as active ingredient, a derivative of formula (I) according to claims 1 to 8, in combination with a pharmaceutical vehicle acceptable for the treatment and / or prevention of ischemical heart disease. 18. Pharmaceutical compositions containing, as active ingredient, a derivative of formula (I) according to claims 1 to 8, in combination with a pharmaceutical vehicle acceptable for the treatment and / or prevention of traumatic or degenerative ischemical pathologies of the system central or peripheral nervous. 19. Pharmaceutical compositions containing, as active ingredient, a derivative of formula (I) according to claims 1 to 8, in combination with a pharmaceutical vehicle acceptable for the treatment and / or prevention of acute or chronic inflammatory diseases. 20. Pharmaceutical compounds containing, as active principle, a derivative of formula (I) according to claims 1 to 8, in combination with a pharmaceutical carrier acceptable for the preventive and curative treatment of certain cancers such as metaplasias and epithelial dysplasias , photo-induced skin cancers (spino-cellular or basal-cell epithelioma) as well as skin precancerous bonds (actinic keratoses).