Silylated derivatives of resveratrol and the use thereof in neurodegenerative, neurological or inflammatory diseases

Abstract

The present invention relates to a group of compounds derived from resveratrol having as substituents at least one silyl group which, in turn, can be substituted by different groups. The invention also relates to the therapeutic use of these compounds in inflammatory, neurological, and neurodegenerative diseases.

Claims

1. A compound of formula (I′): ##STR00034## wherein R.sub.1, R.sub.2, and R.sub.3 are independently selected from the group consisting of H, a SiR.sub.4R.sub.5R.sub.6 group, —NH(CO)R.sub.7 group, and a carbohydrate, and wherein R.sub.4, R.sub.5, and R.sub.6 are independently selected from the group consisting of linear C.sub.1-C.sub.6alkyl, branched C.sub.1-C.sub.6 alkyl and a phenyl group and R.sub.7 is selected from the group consisting of a linear C.sub.1-C.sub.12 alkyl and branched C1-C.sub.12 alkyl, with the proviso that at least one of R.sub.1, R.sub.2, and R.sub.3 is a SiR.sub.4R.sub.5R.sub.6 group and that it is not one of the following compounds: (E)-(5-(4-(trimethylsilyloxy)styryl)-1,3-phenylene)bis(oxy)bis(trimethylsilane), (E)-4-(3,5-bis(triisopropylsilyloxy)styryl)phenol, (E)-3-(tert-butyldimethylsilyloxy)-5-(4-(tert-butyldimethylsilyloxy)styryl)phenol, (E)-4-(3,5-bis(tert-butyldimethylsilyloxy)styryl)phenol, (E)-3-(tert-butyldimethylsilyloxy)-5-(4-hydroxystyryl)phenol, (E)-5-(4-(tert-butyldimethylsilyloxy)styryl)benzene-1,3-diol, (E)-(5-(4-(tert-butyldimethylsilyloxy)styryl)-1,3-phenylene)bis(oxy)bis(tert-butyldimethylsilane), wherein R.sub.1 and R.sub.2 are a SiR.sub.4R.sub.5R.sub.6 group and R.sub.3 is selected from the group consisting of H, —NH(CO)R.sub.7, and the following carbohydrate: ##STR00035## and wherein R.sub.8 is selected from the group consisting of H, and —C(O)—R.sub.9, and R.sub.9 is selected from the group consisting of a C.sub.1-C.sub.22 alkyl and a C.sub.1-C.sub.22 alkenyl, or wherein R.sub.1 and R.sub.3 are a SiR.sub.4R.sub.5R.sub.6 group and R.sub.2 is selected from the group consisting of H, —NH(CO)R.sub.7, and the following carbohydrate: ##STR00036## and wherein R.sub.8 is selected from the group consisting of H, and —C(O)—R.sub.9, and R.sub.9 is selected from the group consisting of a C.sub.1-C.sub.22 alkyl and a C.sub.1-C.sub.22 alkenyl, or wherein R.sub.1, R.sub.2, and R.sub.3 are a SiR.sub.4R.sub.5R.sub.6 group, or wherein the compound is selected from the following group consisting of: ##STR00037## or the compound is selected from the following group consisting of: ##STR00038##

2. The compound of claim 1, wherein R.sub.4 and R.sub.5 are selected from the group consisting of methyl, ethyl, and isopropyl and R.sub.6 is selected from the group consisting of tert-butyl, ethyl, and isopropyl.

3. The compound of formula (I′) according to claim 1, wherein the compound is selected from the following group consisting of: ##STR00039## ##STR00040##

4. A pharmaceutical composition comprising the compound according to claim 1.

Description

BRIEF DESCRIPTION OF THE FIGURES

(1) FIG. 1. Cell viability in neuroblastoma SH-SY5Y after damage with H.sub.2O.sub.2 and treatment with the different compounds 2-9. The controls are DMSO: 1% DMSO; H.sub.2O.sub.2: H.sub.2O.sub.2 in 1% DMSO; RES 10 μM+H.sub.2O.sub.2: resveratrol in H.sub.2O.sub.2 in 1% DMSO.

(2) FIG. 2. Cell viability in neuroblastoma SH-SY5Y after damage with H.sub.2O.sub.2 and treatment with the different compounds 10-11. The controls are DMSO: 1% DMSO; H.sub.2O.sub.2: H.sub.2O.sub.2 in 1% DMSO; RES 10 μM+H.sub.2O.sub.2: resveratrol in H.sub.2O.sub.2 in 1% DMSO.

(3) FIG. 3. Cell viability in neuroblastoma SH-SY5Y after damage with H.sub.2O.sub.2 and treatment with the different compounds 12-13. The controls are DMSO: 1% DMSO; H.sub.2O.sub.2: H.sub.2O.sub.2 in 1% DMSO; RES 10 μM+H.sub.2O.sub.2: resveratrol in H.sub.2O.sub.2 in 1% DMSO.

(4) FIG. 4. Cell viability in neuroblastoma SH-SY5Y after damage with H.sub.2O.sub.2 and treatment with the different compounds 14-15. The controls are DMSO: 1% DMSO; H.sub.2O.sub.2: H.sub.2O.sub.2 in 1% DMSO; RES 10 μM+H.sub.2O.sub.2: resveratrol in H.sub.2O.sub.2 in 1% DMSO.

(5) FIG. 5. Cell viability in neuroblastoma SH-SY5Y after damage with H.sub.2O.sub.2 and treatment with the different compounds 16-20. The controls are DMSO: 1% DMSO; H.sub.2O.sub.2: H.sub.2O.sub.2 in 1% DMSO; RES 10 μM+H.sub.2O.sub.2: resveratrol in H.sub.2O.sub.2 in 1% DMSO.

(6) FIG. 6. Cell viability in RAW macrophages after inflammation produced with LPS and treatment with the different compounds 2-9. The controls are DMSO: 1% DMSO; LPS alone: LPS (100 ng/ml); LPS+RES 10 μM: LPS (100 ng/ml)+resveratrol.

(7) FIG. 7. Cell viability in RAW macrophages after inflammation produced with LPS and treatment with the different compounds 10-11. The controls are DMSO: 1% DMSO; LPS alone: LPS (100 ng/ml); LPS+RES 10 μM: LPS (100 ng/ml)+resveratrol.

(8) FIG. 8. Cell viability in RAW macrophages after inflammation produced with LPS and treatment with the different compounds 12-13. The controls are DMSO: 1% DMSO; LPS alone: LPS (100 ng/ml); LPS+RES 10 μM: LPS (100 ng/ml)+resveratrol.

(9) FIG. 9. Cell viability in RAW macrophages after inflammation produced with LPS and treatment with the different compounds 14-15. The controls are DMSO: 1% DMSO; LPS alone: LPS (100 ng/ml); LPS+RES 10 μM: LPS (100 ng/ml)+resveratrol.

(10) FIG. 10. Cell viability in RAW macrophages after inflammation produced with LPS and treatment with the different compounds 16-20. The controls are DMSO: 1% DMSO; LPS alone: LPS (100 ng/ml); LPS+RES 10 μM: LPS (100 ng/ml)+resveratrol.

(11) FIG. 11. Concentration of TNF-alpha in culture medium after inflammation by LPS in RAW macrophages and treatment with the different compounds 2-9.

(12) FIG. 12. Concentration of NO in culture medium after inflammation by LPS in RAW macrophages and treatment with the different compounds 2-9.

(13) FIG. 13. Concentration of IL6 in culture medium after inflammation by LPS in RAW macrophages and treatment with the different compounds 2-9.

(14) FIG. 14. AChE activity regarding the control of the compounds RES, 6, 8, and 9. An ANOVA statistical test was carried out, followed by Dunnett's multiple comparison test. It is considered significant when #P<0.05 relative to the control; *P<0.05, **P<0.01 relative to the control+PTZ.

(15) FIG. 15. AChE activity regarding the control of the compounds RES, 11, 14, 15, and 17. An ANOVA statistical test was carried out, followed by Dunnett's multiple comparison test. It is considered significant when #P<0.05 relative to the control; *P<0.05, **P<0.01 relative to the control+PTZ.

(16) FIG. 16: Schematic of the design of the experiment conducted in example 6.

(17) FIG. 17: a) Evaluation of the motor capacity (sum of four movement parameters) of the groups of mice treated with the compounds RES and 15; b) Time (in seconds) in the rotarod of the groups of mice treated with the compounds RES and 15 on day 5; c) Average weight of the groups of mice treated with the compounds RES and 15 on day 5; d) Amount of interleukin IL-6 in plasma of the groups of mice treated with the compounds RES and 15.

(18) FIG. 18: a) Evaluation of the motor capacity of the groups of mice treated preventively with compound 15 and the EAE-T control; b) Evaluation of the motor capacity of the groups of mice treated preventively with the compound RES and the EAE control without T; c) Evaluation of the motor capacity of the groups of mice treated therapeutically with compound 15 and the EAE-T control; d) Evaluation of the motor capacity of the groups of mice treated therapeutically with the RES compound and the EAE control without T.

EXAMPLES

(19) The invention will be illustrated below by means of tests conducted by the inventors that demonstrate the effectiveness of the product of the invention.

Example 1: Synthesis of the Silylated Derivatives of Resveratrol

(20) ##STR00013##
General Method of Silylation.

(21) Resveratrol (1 eq.) and imidazole (2.5 eq.) were added to a round-bottom flask with DMF (3 ml/mmol resveratrol) under stirring and cooled to 0° C. The corresponding silyl chloride (1, 4-1, 55 eq.) was then added dropwise in two batches, the first half at=0 h and the second half at=3 h. The reaction was stirred for a total of 6 h at room temperature. The reaction mixture was then filtered, diluted in water, and extracted with ethyl acetate (3×20 ml). The combined organic phases were dried with MgSO.sub.4, filtered, concentrated to dryness, and purified by silica gel column chromatography, with elution being performed with hexane/ethyl acetate mixtures.

(22) Series of Tert-Butyldimethylsilyl Resveratrol Derivatives.

(23) Following the general method, resveratrol (830 mg, 3.64 mmol) and tert-butyldimethylsilyl chloride (849.55 mg, 5.64 mmol) were used to obtain compounds 1, 2, and 3 in addition to the monosilylated derivatives after purifying the reaction mixture by column chromatography using a gradient of hexane/ethyl acetate (8:1 to 2:1).

(24) ##STR00014##

(25) 3,4′,5-O-tri-tert-butyldimethylsilyl resveratrol, compound 1. Yield=5.8%; R.sub.f=0.9 (hexane:ethyl acetate—7:3).

(26) ##STR00015##

(27) 3,4′,-O-tri-tert-butyldimethylsilyl resveratrol, compound 2. Yield=22.1%; R.sub.f=0.65 (hexane:ethyl acetate—7:3).

(28) ##STR00016##

(29) 3,5,-O-tri-tert-butyldimethylsilyl resveratrol, compound 3. Yield=6.4%; R.sub.f=0.55 (hexane:ethyl acetate—7:3).

(30) Series of Triisopropylsilyl Resveratrol Derivatives.

(31) Following the general method, resveratrol (816 mg, 3.57 mmol) and triisopropylsilyl chloride (1.19, 5.54 mmol) were used to obtain compounds 4, 5, and 6 in addition to the monosilylated derivatives after purifying the reaction mixture by column chromatography using a gradient of hexane/ethyl acetate (10:1 to 1:1).

(32) ##STR00017##

(33) 3,4′,5-O-tri-triisopropylsilyl resveratrol, compound 4. Yield=19.4%; R.sub.f=0.9 (hexane:ethyl acetate—3:1). RMN of .sup.1H (400 MHz, CDCl.sub.3): δ=7.47 (d, J=8.3 Hz, 2H), 7.05 (d, J=16.2 Hz, 1H, CH), 6.95 (dd, J=12.4, 6.4 Hz, 3H, CH and H), 6.74 (s, 2H, H.sup.2 and H.sup.6), 6.44 (s, 1H, H.sup.4), 1.41-1.33 (m, 9H, CH—Si), 1.29-1.16 (m, 54H, CH.sub.3). RMN of .sup.13C (101 MHz, CDCl.sub.3): δ=157.10, 155.96, 139.44, 130.45, 128.39, 127.73, 126.83, 120.17, 111.26, 110.94, 18.02, 12.78.

(34) ##STR00018##

(35) 3,4′,-O-di-triisopropylsilyl resveratrol, compound 5. Yield=26.1%; R.sub.f=0.65 (hexane:ethyl acetate—3:1). RMN of .sup.1H (400 MHz, CDCl.sub.3): b=7.34 (d, J=8.2 Hz, 2H), 6.96 (d, J=16.2 Hz, 1H), 6.83 (t, J=12.0 Hz, 3H), 6.64 (s, 1H), 6.55 (s, 1H), 6.33 (s, 1H), 1.30-1.22 (m, 6H), 1.11 (dd, J=16.0, 7.4 Hz, 36H). RMN of .sup.13C (101 MHz, CDCl.sub.3): δ=158.14, 157.12, 155.68, 139.63, 130.56, 127.98, 127.47, 126.57, 119.77, 109.57, 106.15, 106.06, 17.40, 17.37, 17.10, 12.62, 12.59, 12.32. TOF MS-ES, calculated mass: C.sub.32H.sub.51O.sub.3Si.sub.2 [M−H]=539.3377, measured mass: [M−H]=539.3390.

(36) ##STR00019##

(37) 3,5,-O-di-triisopropylsilyl resveratrol, compound 6. Yield=8.7%; R.sub.f=0.5 (hexane:ethyl acetate—3:1).

(38) Series of Triethylsilyl Resveratrol Derivatives.

(39) Following the general method, resveratrol (809 mg, 3.54 mmol) and ethylsilyl chloride (1.19, 5.49 mmol) were used to obtain compounds 7, 8, and 9 in addition to the monosilylated derivatives after purifying the reaction mixture by column chromatography using a gradient of hexane/ethyl acetate (10:1 to 1:1).

(40) ##STR00020##

(41) 3,4′,5-O-tri-triethylsilyl resveratrol, compound 7. Yield=3.8%; R.sub.f=0.95 (hexane:ethyl acetate—3:1). RMN of .sup.1H (400 MHz, CDCl.sub.3): δ=7.40 (d, J=8.5 Hz, 2H), 6.98 (d, J=16.2 Hz, 1H), 6.90-6.82 (m, 3H), 6.64 (d, J=2.0 Hz, 2H), 6.31 (t, J=2.0 Hz, 1H), 1.04 (td, J=7.8, 2.8 Hz, 26H), 0.78 (q, J=7.9 Hz, 18H). RMN of .sup.13C (101 MHz, CDCl.sub.3): =156.61, 155.45, 139.48, 130.60, 128.44, 127.73, 126.76, 120.17, 111.41, 110.93, 6.66, 5.03. Calculated mass: C.sub.32H.sub.55O.sub.3Si.sub.3 [M+H]=571.3459, measured mass: [M+H]=571.3460.

(42) ##STR00021##

(43) 3,4′,-O-tri-triethylsilyl resveratrol, compound 8. Yield=14.9%; R.sub.f=0.6 (hexane:ethyl acetate—3:1). RMN of .sup.1H (400 MHz, CDCl.sub.3): δ=7.40 (d, J=8.6 Hz, 2H), 6.98 (d, J=16.2 Hz, 1H), 6.91-6.82 (m, 3H), 6.62 (s, 1H), 6.61 (s, 1H), 6.32 (s, 1H), 1.09-1.02 (m, 18H), 0.80 (q, J=7.9 Hz, 12H). RMN of .sup.13C (101 MHz, CDCl.sub.3): δ=156.92, 156.69, 155.45, 139.94, 130.58, 128.82, 127.83, 126.47, 120.25, 110.96, 106.52, 106.40, 6.67, 6.64, 5.03. TOF MS-ES.sup.+, calculated mass: C.sub.26H.sub.41O.sub.3Si.sub.2 [M+H]=457.2594, measured mass: [M+H]=457.2593.

(44) ##STR00022##

(45) 3,5,-O-tri-triethylsilyl resveratrol, compound 9. Yield=6.8%; R.sub.f=0.5 (hexane:ethyl acetate—3:1). RMN of .sup.1H (400 MHz, CDCl.sub.3): 7.37 (d, J=8.3 Hz, 2H), 6.96 (d, J=16.2 Hz, 1H), 6.82 (t, J=12.6 Hz, 3H), 6.63 (s, 2H), 6.25 (s, 1H), 1.02 (t, J=7.9 Hz, 18H), 0.76 (q, J=7.9 Hz, 12H). RMN of .sup.13C (101 MHz, CDCl.sub.3): δ=157.11, 156.55, 140.04, 128.84, 128.66, 127.64, 127.55, 125.16, 115.19, 110.99, 110.08, 48.34, 48.13, 47.92, 47.70, 47.49, 47.28, 47.06, 5.77, 4.66. TOF MS-ES.sup.+, calculated mass: C.sub.26H.sub.41O.sub.3Si.sub.2 [M+H]=457.2594, measured mass: [M+H]=457.2586.

(46) Series of Triisopropylsilyl and Ethyl Carbamide Resveratrol Derivatives.

(47) Ethyl isocyanate (1.5 eq.) and triethylamine (2 eq.) were added to a solution of 3,4′-dithiisopropylsilyl resveratrol or 3,5-dithiisopropylsilyl resveratrol (1 eq.) in dichloromethane. After 1 h of reaction at room temperature, the reaction was concentrated and purified on a chromatography column, with elution being performed with hexane:ethyl acetate (from 2:1 to 0:1).

(48) ##STR00023##

(49) 3,4′,-O-ditriisopropylsilyl-5-ethyl carbamate resveratrol, compound 10. Yield=80.2%. R.sub.f=0.9 (hexane:ethyl acetate—5:1). RMN of .sup.1H (300 MHz, CDCl.sub.3): δ=7.39 (d, J=8.5 Hz, 2H), 7.02 (d, J=16.2 Hz, 1H), 6.89 (dd, J=14.6, 5.9 Hz, 5H), 6.60 (s, 1H), 3.40-3.26 (m, 2H), 1.30 (ddd, J=10.6, 7.4, 3.7 Hz, 9H), 1.15 (dd, J=7.0, 3.1 Hz, 36H). RMN of .sup.13C (75 MHz, CDCl.sub.3): δ=157.04, 156.29, 152.30, 139.75, 130.38, 129.38, 128.01, 126.25, 120.39, 115.12, 112.30, 36.38, 18.21, 18.18, 17.97, 15.40, 12.95, 12.93, 12.57. TOF MS-ES.sup.+, calculated mass: C.sub.35H.sub.57NO.sub.4Si.sub.2 [M+H]=612.3904, measured mass: [M+H]=612.3907.

(50) ##STR00024##

(51) 3,5-O-ditriisopropylsilyl-4-ethyl carbamate resveratrol, compound 11. Yield=85.6%. R.sub.f=0.9 (hexane:ethyl acetate—5:1). RMN of .sup.1H (300 MHz, CDCl.sub.3): δ=7.51 (d, J=8.0 Hz, 2H), 7.15 (d, J=8.0 Hz, 2H), 7.01 (d, J=16.4 Hz, 1H), 6.93 (d, J=16.1 Hz, 1H), 6.67 (s, 2H), 6.38 (s, 1H), 3.41-3.28 (m, 2H), 1.32-1.22 (m, 9H), 1.15 (d, J=7.2 Hz, 36H). RMN of .sup.13C (75 MHz, CDCl.sub.3): δ=157.31, 157.25, 139.15, 128.14, 128.00, 127.59, 121.99, 116.20, 116.04, 112.17, 111.61, 111.38, 45.95, 36.41, 18.20, 12.95. TOF MS-ES.sup.+, calculated mass: C.sub.35H.sub.57NO.sub.4Si.sub.2 [M+H]=612.3904, measured mass: [M+H]=612.3900.

(52) Series of Triisopropylsilyl Glucosyl Resveratrol Derivatives.

(53) Under stirring and an inert argon atmosphere, 3,4′-ditriisopropylsilyl resveratrol or 3,5-ditriisopropylsilyl resveratrol (1 eq.) was dissolved in 15 ml of anhydrous dichloromethane and peracetyl glucose trifluoroacetimidate (1.5 eq.) and boron trifluoride etherate (0.1 eq.) were added. After 30 min of reaction, 5 ml of triethylamine was added, concentrated, and added to a silica gel purification column, with elution being performed with a mixture of hexane and ethyl acetate (5:1). The obtained product was dissolved in a mixture of dichloromethane, water, and methanol (5 ml, 2:1:2), and sodium bicarbonate (3 eq.) was added. After deprotection of the acetate units of the glucose unit (24-48 h), the reaction was concentrated and purified by column chromatography, with elution being performed with hexane:ethyl acetate (from 1:1 to 1:3).

(54) ##STR00025##

(55) 3,4′-O-ditriisopropylsilyl-5-glucosyl resveratrol, compound 12. Yield=70.6%. R.sub.f=0.05 (hexane:ethyl acetate—1:3). RMN of .sup.1H (500 MHz, CD.sub.3OD): δ=7.43 (d, J=8.6 Hz, 2H), 7.05 (d, J=16.3 Hz, 1H), 6.98-6.89 (m, 2H), 6.87 (d, J=8.6 Hz, 2H), 6.70 (s, 1H), 6.55 (t, J=2.0 Hz, 1H), 4.92-4.88 (m, 1H), 3.94-3.88 (m, 1H), 3.75 (dd, J=11.9, 4.8 Hz, 1H), 3.52-3.38 (m, 4H), 1.36-1.23 (m, 6H), 1.18-1.10 (m, 36H). RMN of .sup.13C (126 MHz, CD.sub.3OD): δ=158.94, 156.93, 155.80, 139.83, 130.51, 128.55, 127.51, 126.04, 119.69, 111.96, 107.30, 107.07, 101.13, 76.86, 76.59, 73.48, 69.90, 61.01, 17.05, 16.99, 12.53, 12.51. TOF MS-ES.sup.+, calculated mass: C.sub.38H.sub.62O.sub.8Si.sub.2 [M+Na]=725.3881, measured mass: [M+Na]=725.3682.

(56) ##STR00026##

(57) 3,5-O-ditriisopropylsilyl-4′-glucosyl resveratrol, compound 13. Yield=75.3%. R.sub.f=0.05 (hexane:ethyl acetate—1:3). RMN of .sup.1H (500 MHz, CD.sub.3OD): 5=7.48 (d, J=8.7 Hz, 2H), 7.09 (d, J=8.7 Hz, 2H), 6.95 (q, J=16.3 Hz, 3H), 6.66 (d, J=2.0 Hz, 2H), 6.32 (t, J=2.1 Hz, 1H), 4.96-4.90 (m, 1H), 3.91 (dd, J=12.1, 1.9 Hz, 1H), 3.73 (dd, J=12.1, 5.3 Hz, 1H), 3.51-3.47 (m, 2H), 3.45 (dd, J=5.0, 1.7 Hz, 1H), 3.44-3.38 (m, 1H), 1.31-1.23 (m, 6H), 1.14 (d, J=7.3 Hz, 36H). RMN of .sup.13C (126 MHz, CD.sub.3OD): δ=157.41, 156.96, 139.63, 131.54, 128.05, 127.37, 126.65, 116.54, 110.93, 110.28, 100.83, 76.75, 76.57, 73.49, 69.94, 61.10, 17.05, 12.55. TOF MS-ES.sup.+, calculated mass: C.sub.38H.sub.62O.sub.8Si.sub.2 [M+Na]=725.3881, measured mass: [M+Na]=725.3907.

(58) Series of Triisopropylsilyl Octanoyl-Glucosyl Resveratrol Derivatives.

(59) 3,4′-O-ditriisopropylsilyl-5-glucosyl resveratrol or 3,5-O-dithiisopropylsilyl-4′-glucosyl resveratrol (1 eq.) was dissolved in methyl tert-butyl ether and vinyl octanoate (3 eq.) and the enzyme Lypozyme TL IM® (same amount in grams as the resveratrol derivative). After 3 days of reaction, the enzyme was filtered and washed with ethyl acetate and methanol. After concentration of the solvent, it was purified by chromatography on a silica column, with elution being performed with hexane:ethyl acetate (from 2:1 to 1:3).

(60) ##STR00027##

(61) 3,4′-O-ditriisopropylsilyl-5-(-6-octanoyl)glucosyl resveratrol, compound 14. Yield=75.3%. R.sub.f=0.05 (hexane:ethyl acetate—1:3). RMN of .sup.1H (500 MHz, CDCl.sub.3): =.sup.1H NMR (500 MHz, CdCl.sub.3) δ=7.37 (d, J=8.5 Hz, 2H), 7.12 (d, J=8.5 Hz, 1H), 6.98 (d, J=16.1 Hz, 2H), 6.87 (d, J=8.5 Hz, 2H), 6.75 (s, 1H), 6.73 (s, 1H), 4.94-4.87 (m, 1H), 4.75-4.70 (m, 1H), 4.59-4.48 (m, 2H), 3.69-3.60 (m, 3H), 1.28-1.25 (m, 18H), 1.15-1.07 (m, 36H), 0.91-0.89 (m, 3H). RMN of .sup.13C (126 MHz, CDCl.sub.3): δ=171.07, 139.75, 130.16, 127.72, 126.19, 120.16, 119.68, 115.09, 112.90, 109.58, 107.50, 106.00, 104.78, 102.13, 100.89, 66.80, 60.38, 45.71, 38.73, 34.00, 31.91, 30.40, 29.69, 29.35, 28.91, 24.47, 23.78, 22.68, 21.03, 20.82, 17.94, 17.89, 17.86, 14.18, 14.10, 12.67, 12.62, 12.54, 10.96, 8.76. TOF MS-ES−, calculated mass: C.sub.46H.sub.76O.sub.9Si.sub.2 [M−H]=827.4950, measured mass: [M−H]=827.4922.

(62) ##STR00028##

(63) 3,5-O-ditriisopropylsilyl-4′-(6-octanoyl)glucosyl resveratrol, compound 15. Yield=15.5%. R.sub.f=0.07 (hexane:ethyl acetate—1:2). RMN of .sup.1H (300 MHz, CDCl.sub.3): δ 7.45 (d, J=8.5 Hz, 2H), 7.07 (d, J=8.4 Hz, 2H), 6.93 (q, J=16.2 Hz, 2H), 6.66 (s, 2H), 6.37 (s, 1H), 4.96 (d, J=6.0 Hz, 1H), 4.43 (m, 2H), 3.77-3.55 (m, 4H), 2.36 (t, J=7.5 Hz, 2H), 1.63 (d, J=6.8 Hz, 2H), 1.29-1.24 (m, 8H), 1.14 (d, J=7.0 Hz, 36H), 0.92-0.84 (m, 3H). RMN of .sup.13C (75 MHz, CDCl.sub.3): δ=174.78, 157.28, 139.28, 132.63, 127.93, 117.30, 111.52, 74.32, 73.59, 34.47, 31.88, 29.97, 29.36, 29.17, 25.12, 22.84, 18.18, 14.29, 12.94. TOF MS-ES.sup.+, calculated mass: C.sub.46H.sub.76O.sub.9Si.sub.2 [M+Na]=851.4926, measured mass: [M+Na]=851.4966.

(64) Series of Triethylsilyl Resveratrol Derivatives with Acyl Groups.

(65) 3,5-O-triethylsilyl resveratrol (9) (1 eq.) was dissolved in tert-butanol, and the vinyl ester of the corresponding fatty acid (6 eq.) and Novozyme 435® (approx. 100 mg) were added. The reaction was allowed to progress for 60 h at 50° C. under orbital shaking. After this period, the reaction was filtered in order to remove the enzyme and washed with a little methanol. The obtained crude was purified by means of silica gel column chromatography with a gradient using a mixture of hexane and ethyl acetate as the mobile phase (100:0-1:1).

(66) ##STR00029##

(67) 3,5-O-triethylsilyl-4′-propanoyl resveratrol, compound 16. Yield=34.0%. R.sub.f=0.35 (hexane:ethyl acetate—8:1). RMN of .sup.1H (400 MHz, CDCl.sub.3): δ=7.37 (d, J=8.3 Hz, 2H), 6.94 (d, J=16.2 Hz, 1H), 6.87-6.73 (m, 3H), 6.60 (d, J=2.3 Hz, 2H), 6.28 (t, J=2.1 Hz, 1H), 2.33 (q, 2H), 1.66 (t, 3H), 1.01 (t, J=7.9 Hz, 18H), 0.75 (q, J=7.9 Hz, 12H). RMN of .sup.13C (101 MHz, CDCl.sub.3): δ=180.10 (CO), 156.55 (2×Cq), 155.26 (Cq), 139.39 (Cq), 130.20 (Cq), 128.24 (CH arom), 127.91 (2×CH arom), 126.60 (CH arom), 115.56 (2×CH arom), 111.38 (2×CH arom), 110.93 (CH arom), 29.67 (CH.sub.2CO), 14.06 (CH.sub.3CH.sub.2CO), 6.60 (6×CH.sub.2Si), 5.00 (6×CH.sub.3CH.sub.2Si).

(68) ##STR00030##

(69) 3,5-O-triethylsilyl-4′-butanoyl resveratrol, compound 17. Yield=39.0%. R.sub.f=0.29 (hexane:ethyl acetate—8:1). RMN of .sup.1H (400 MHz, CDCl.sub.3): δ=7.37 (d, J=8.0 Hz, 2H), 6.93 (d, J=16.2 Hz, 1H), 6.87-6.76 (m, 3H), 6.60 (d, J=2.2 Hz, 2H), 6.27 (t, J=2.1 Hz, 1H), 2.34 (t, 2H), 1.66 (m, 2H), 1.00 (t, J=7.9 Hz, 18H), 0.94 (t, 3H), 0.74 (q, J=7.9 Hz, 12H). RMN of .sup.13C (101 MHz, CDCl.sub.3): δ=179.26 (CO), 156.55 (2×Cq), 155.32 (Cq), 139.40 (Cq), 130.14 (Cq), 128.25 (CH arom), 127.91 (2×CH arom), 126.56 (CH arom), 115.57 (2×CH arom), 111.38 (2×CH arom), 110.91 (CH arom), 29.65 (CH.sub.2CO), 22.66 (CH.sub.2CH.sub.3), 14.06 (CH.sub.3), 6.60 (6×CH.sub.2Si), 4.99 (6×CH.sub.3CH.sub.2Si).

(70) ##STR00031##

(71) 3,5-O-triethylsilyl-4′-hexanoyl resveratrol, compound 18. Yield=66.0%. R.sub.f=0.29 (hexane:ethyl acetate—8:1). RMN of .sup.1H (400 MHz, CDCl.sub.3): δ=7.36 (d, J=8.0 Hz, 2H), 6.93 (d, J=16.2 Hz, 1H), 6.85-6.76 (m, 3H), 6.59 (d, J=2.2 Hz, 2H), 6.26 (t, J=2.2 Hz, 1H), 2.33 (t, J=7.5 Hz, 2H), 1.70-1.56 (m, 2H), 1.32 (m, 2H), 1.00 (t, J=7.9 Hz, 18H), 0.92-0.84 (m, 3H), 0.79-0.69 (m, 12H). RMN of .sup.13C (101 MHz, CDCl.sub.3): δ=180.26 (CO), 156.54 (2×Cq), 155.39 (Cq), 139.41 (Cq), 130.07 (Cq), 128.28 (CH arom), 127.88 (2×CH arom), 126.49 (CH arom), 115.57 (2×CH arom), 111.36 (2×CH arom), 110.88 (CH arom), 34.02 (CH.sub.2CO), 31.16 (CH.sub.2CH.sub.2), 24.33 (CH.sub.2CH.sub.2), 22.24 (CH.sub.2CH.sub.3), 13.80 (CH.sub.3), 6.58 (6×CH.sub.2Si), 4.98 (6×CH.sub.3CH.sub.2Si).

(72) ##STR00032##

(73) 3,5-O-triethylsilyl-4′-octanoyl resveratrol, compound 19. Yield=63.0%. R.sub.f=0.25 (hexane:ethyl acetate—8:1). RMN of .sup.1H (400 MHz, CDCl.sub.3): δ=7.36 (d, J=8.3 Hz, 2H), 6.93 (d, J=16.3 Hz, 1H), 6.87-6.72 (m, 3H), 6.59 (d, J=2.2 Hz, 2H), 6.26 (t, J=2.2 Hz, 1H), 2.33 (t, J=7.5 Hz, 2H), 1.62 (q, J=7.4 Hz, 2H), 1.29 (m, 8H), 1.00 (t, J=7.9 Hz, 18H), 0.86 (t, J=6.6 Hz, 3H), 0.74 (q, J=7.9 Hz, 12H). RMN of .sup.13C (101 MHz, CDCl.sub.3): δ=180.38 (CO), 156.54 (2×Cq), 155.43 (Cq), 139.40 (Cq), 130.04 (Cq), 128.27 (CH arom), 127.87 (2×CH arom), 126.48 (CH arom), 115.56 (2×CH arom), 111.34 (2×CH arom), 110.87 (CH arom), 34.07 (CH.sub.2CO), 31.58 (CH.sub.2CH.sub.2), 28.97 (CH.sub.2CH.sub.2), 28.85 (CH.sub.2CH.sub.2), 24.64 (CH.sub.2CH.sub.2), 22.54 (CH.sub.2CH.sub.3), 13.98 (CH.sub.3), 6.58 (6×CH.sub.2Si), 4.98 (6×CH.sub.3CH.sub.2Si).

(74) ##STR00033##

(75) 3,5-O-triethylsilyl-4′-decanoate resveratrol, compound 20. Yield=89.0%. R.sub.f=0.25 (hexane:ethyl acetate—8:1). RMN of .sup.1H (400 MHz, CDCl.sub.3): δ=7.36 (d, J=8.2 Hz, 2H), 6.93 (d, J=16.2 Hz, 1H), 6.88-6.74 (m, 3H), 6.59 (d, J=2.2 Hz, 2H), 6.26 (t, J=2.2 Hz, 1H), 2.33 (t, J=7.5 Hz, 2H), 1.62 (q, J=7.4 Hz, 2H), 1.41-1.18 (m, 12H), 1.00 (t, J=7.9 Hz, 18H), 0.86 (t, J=6.8 Hz, 3H), 0.74 (q, J=7.8 Hz, 12H). RMN of .sup.13C (101 MHz, CDCl.sub.3): δ=180.43 (CO), 156.54 (2×Cq), 155.48 (Cq), 139.42 (Cq), 130.00 (Cq), 128.30 (CH arom), 127.86 (2×CH arom), 126.44 (CH arom), 115.57 (2×CH arom), 111.34 (2×CH arom), 110.86 (CH arom), 34.08 (CH.sub.2CO), 31.82 (CH.sub.2CH.sub.2), 29.35 (CH.sub.2CH.sub.2), 29.21 (CH.sub.2CH.sub.2), 29.20 (CH.sub.2CH.sub.2), 29.02 (CH.sub.2CH.sub.2), 24.64 (CH.sub.2CH.sub.2), 22.62 (CH.sub.2CH.sub.3), 14.02 (CH.sub.3), 6.57 (6×CH.sub.2Si), 4.98 (6×CH.sub.3CH.sub.2Si).

Example 2: Viability and Neuroprotection Assays

(76) The SH-SY5Y neuroblastoma cell line was cultured in petri dishes pre-treated with collagen (100 ng/ml) with F12 medium supplemented with penicillin/streptomycin and 10% inactivated fetal bovine serum.

(77) Cell viability assays with neurons were prepared in 96-well plates pre-treated with collagen by seeding 20,000 cells/well in a volume of 100 μL and incubating the cells for 24 h before the addition of the compounds. The compounds to be tested were dissolved in DMSO and added in three different concentrations (1, 10, and 100 μM) in order to determine their toxicity. The final percentage of DMSO in each well was adjusted to 1%. The cell viability was evaluated 24 h after the addition of the compounds by means of the MTT assay according to the manufacturer's method. Mean values and standard deviations were calculated from at least eight different measurements from several independent experiments.

(78) For the neuroprotection assay, the neurons were cultured and seeded in the same manner as for the cell viability assay. The compounds to be tested were dissolved in DMSO and added in three different concentrations (1, 10, and 100 μM) and, after 10-minute incubation, hydrogen peroxide (100 μM) was added to the medium. The final percentage of DMSO in each well was adjusted to 1%. The cell viability was evaluated 24 h after the addition of the compounds by means of the MTT assay according to the manufacturer's method. Mean values and standard deviations were calculated from at least eight different measurements from several independent experiments. Neuronal recovery was calculated by normalizing the results of the neuronal viability experiments after the addition of the compounds of the invention and H.sub.2O.sub.2 to the positive control of each experiment (neurons+H.sub.2O.sub.2).

(79) It is observed that the RES 10 μM control recovers up to 50% of cell viability (FIGS. 1 to 5, indicated by the broken line). In contrast, many of the silylated derivatives of the invention recover between 80 and 120% viability at concentrations between 1 and 100 μM. Some appear to exhibit toxicity at 100 μM.

Example 3: Inflammation Assay

(80) RAW 264.7 macrophages were cultured in P75 with high-glucose DMEM supplemented with penicillin/streptomycin and 10% inactivated fetal bovine serum.

(81) The cell viability assays with RAW macrophages were prepared in 96-well plates by seeding 25,000 cells/well in a volume of 100 μL and incubating the cells for 4 h before the addition of the compounds. The compounds to be tested were dissolved in DMSO and added in three different concentrations (1, 10, and 100 μM) in order to determine their toxicity. The final percentage of DMSO in each well was adjusted to 1%. The cell viability was evaluated 24 h after the addition of the compounds by means of the MTT assay according to the manufacturer's method. Mean values and standard deviations were calculated from at least eight different measurements from several independent experiments.

(82) For the testing of mitigation of damage caused by the addition of LPS, the RAW 264.7 macrophages were cultured according to the procedure described above. The compounds to be tested were dissolved in DMSO and added in three different concentrations (1, 10, and 100 μM) and, after 10-minute incubation, LPS (100 ng/ml) was added to the medium. The final percentage of DMSO in each well was adjusted to 1%. The cell viability was evaluated 24 h after the addition of the compounds by means of the MTT assay according to the manufacturer's method. Mean values and standard deviations were calculated from at least eight different measurements from several independent experiments.

(83) In this assay, it is observed that the 10 μM resveratrol control recovers up to 62% of cell viability (FIGS. 6 to 10, indicated by the broken line). In contrast, several of the silylated derivatives of the invention recover greater cell viability at concentrations between 1 and 100 μM. Some appear to exhibit toxicity at 100 μM.

Example 4: Measurements of Inflammation Parameters in Assay with LPS

(84) To determine the production of cytokines, 5×10.sup.5 RAW 264.7 macrophages were seeded in 24-well plates (0.5 ml per well). The compounds to be tested were then added (10 μM), and the macrophages were either stimulated or not through the addition of LPS (1 μg/ml) to the culture medium. After 24 hours, the levels of IL-6 and TNF-α were measured in the supernatants by ELISA using the capture and biotinylated antibodies from BD PharMingen and PrepoTech following known protocols. The levels of NO in the supernatants at 24 hours were measured indirectly by determining the nitrite concentration in the medium using the Griess reagent according to established protocol. A minimum of two independent experiments and three replicates per experiment were performed for each measured value. The values are expressed as the mean±standard deviation.

(85) In the previous assay, the levels of various inflammation parameters were measured (TNF-α, NO, and IL-6) by ELISA after treatment with RES or with some of the compounds of the invention (2, 3, 5, 6, 8, and 9).

(86) It is observed that the control of RES 10 μM significantly decreases inflammatory parameters (TNF-α, NO, and IL-6) (FIGS. 11, 12, and 13, respectively, indicated by the bar in bold). In contrast, several of the silylated derivatives of the invention improve RES and decrease these parameters even further.

Example 5: Evaluation of the Neuroprotective Capacity of Various Silylated Compounds in a Model of Neurodecqeneration in Zebrafish Larvae Induced by Pentylenetetrazole (PTZ)

(87) The objective of this assay was to analyze the protective effect of various derivatives of resveratrol in a model of neurotoxicity induced by the neurotoxin pentylenetetrazole (PTZ). As an experimental model, the zebrafish (Danio rerio) was used to study the effect of the compounds on acetylcholinesterase activity (AChE) in larvae at 5 days post-fertilization (dpf).

(88) Studies of the central nervous system (CNS) in zebrafish show that, at 24 hours of development, the brain of the embryo has already segmented and already has some structures such as the neural tube, the notochord, and the somites (muscle, and bone precursors). At 5 days post-fertilization (5 dpf), the animal has formed sensory organs such as eyes and otoliths. In addition, the heart, liver, kidneys, and pancreas, as well as the circulatory, digestive, and nervous systems, are fully functional. At this time, the animal is able to respond to visual, olfactory, and mechanical stimuli and begins the search for food.

(89) Zebrafish embryos were seeded in 50 ml of dilution water (AD) in a Petri dish and grown to 5 dpf (larval stage). Only those larvae that did not exhibit any type of external anomaly were used to perform the assay. Next, the larvae were transferred using a Pasteur pipette to a 24-well microplate, so that each well contained five larvae, making ten replicates per condition. First, the pre-treatment of the 5 dpf larvae was performed. For this, the larvae were incubated at 26±1° C. for 1 hour in a volume of 2 ml of AD for the two control groups (Control and Control+PTZ), of physostigmine (Phys) 20 μM, which is a commercial inhibitor of the enzyme AChE for the Phys group, and of the test compounds at a concentration of 10 μM. A medium exchange was then carried out, and the larvae were incubated with the compounds in combination with 5 mM PTZ for 6 hours at 26±1° C. After this incubation period, all of the larvae were examined, and it was determined that the general state of the larvae was totally normal, without any visible anomaly or anomalous behavior. Finally, the larvae were processed for the analysis of the AChE activity.

(90) In order to determine the AChE levels, larvae processing was carried out according to the established technical study protocol once the experimental period was completed. The larvae were homogenized mechanically, and the samples were centrifuged to obtain the supernatant, which were used to determine the levels of the AChE enzyme as a function of the treatments administered. In addition, the determination of total protein of each experimental group was carried out according to the established technical study protocol. Finally, the AChE levels determined in the control group were taken as a reference measurement and deemed to be 100%.

(91) The results of this assay showed that silylated derivatives 9 (3,5-dithyrylsilyl resveratrol) and 15 (3,5-O-ditriisopropylsilyl-4′-(6-octanaoyl)glucosyl resveratrol) significantly prevent the decrease in AChE activity induced by PTZ in 5 dpf larvae, exhibiting a clear neuroprotective effect (see FIGS. 14 and 15). Silylated derivatives 8, 14, and 17 show a lower neuroprotective effect similar to that observed for resveratrol (RES).

Example 6: Testing of Compound 15 in Animal Model of Huntington's Disease

(92) Compound 15 was investigated as a possible treatment in a mouse model of Huntington's disease. The compound resveratrol (RES) was added as a reference.

(93) In this model, increasing amounts of 3-nitro-propionic acid (3NP) are injected into the mice, causing lesions with a phenotype very similar to that of Huntington's disease, both mechanically and pathologically. Once the damage was caused, the mice were treated with RES, compound 15, or vehicle, and we studied the effect thereof on the mouse on the basis of different parameters. Behavioral studies (rotarod test), an assessment was made of motor capacity (tabulated on a scale of 0 normal to 4 incapable, in several parameters such as general mouse dystonia, ability to flip over, and the ability to explore), weight was monitored, and subsequent measurements were taken such as of the inhibition of pro-inflammatory cytokines in plasma or brain.

(94) The design of the experiment is shown in FIG. 16. #=Behavioral study and rotarod test; ϕ Administration of the compound; W=weeks of age of the mice; D=Day of the experiment.

(95) The results are shown in FIG. 17. In all cases, it is observed how compound 15 is capable of improving the phenotype caused by 3NP either by decreasing the severity of the motor lesions or the levels of pro-inflammatory cytokines such as IL-6, or by increasing the weight of the animals. The differences between compound 15 and RES are not great, probably because this animal model is short and very aggressive. It would be advisable to test compound 15 in a model in which the neuronal, motor, and inflammatory damage was slower and more progressive, because it would be closer to the reality of the patient and because the treatment is perhaps much more efficient in a more progressive model.

Example 7: Testing of Compound 15 in Animal Model of Multiple Sclerosis

(96) Compound 15 was investigated as a possible treatment in a mouse model of multiple sclerosis. The compound resveratrol (RES) was added as a reference. An experimental allergic encephalomyelitis model, EAE, was used as an animal model of multiple sclerosis.

(97) In this model, mice are injected with myelin oligodendrocyte glycoprotein (MOG, MOG.sub.35-55, is the 21 amino acid peptide corresponding to the sequence from positions 35 to 55 of the MOG glycoprotein) and virus pertussis (pertussis toxin, PTX) in order to induce the disease. Two days after induction, a new dose of the virus was given again, because some mice were not developing symptoms of EAE. Once the damage was caused, the mice were treated with RES, compound 15, or vehicle, and we studied the effect thereof on the mouse on the basis of different motor parameters on a scale from 0 normal to 4 incapable.

(98) The design of the experiment is shown below:

(99) Experimental Groups:

(100) Vehicle without tween-80 emulsifier (control EAE without T) Vehicle with tween-80 emulsifier (EAE-T control) Preventive RES (without tween-80 emulsifier) (RES-PREV) Therapeutic RES (no tween-80 emulsifier) (RES-TERAP) Preventive compound 15 (with tween-80 emulsifier) (compound 15-PREV) Therapeutic compound 15 (with tween-80 emulsifier) (compound 15-TERAP)

(101) Preventive Regimen (Pretreatment. PREV):

(102) The (intraperitoneal) administration of the compounds (resveratrol, compound 15, and vehicle, with or without tween-80) is started 5 days after induction. 250 μL of the compound in question (20 mg/kg) were injected twice a week and the treatment was maintained for 3 weeks. The treatment was then stopped, but the state of the mice was monitored for another 3 weeks (47 days total).

(103) Therapeutic Regimen (Therapeutic. TERAP):

(104) The (intraperitoneal) administration of the compounds (resveratrol, compound 15, and vehicle, with or without tween-80) is started 12 days after induction. 250 μL of the compound in question (20 mg/kg) were injected twice a week and the treatment was maintained for 2 weeks. The treatment was then stopped, but the state of the mice was monitored for another 3 weeks (47 days total).

(105) When performing a preventive treatment (see FIGS. 18a and 18b), it is observed that compound 15 does not have a great effect on motor capacity (a). On the other hand, RES does reduce damage on a motor level with this preventive treatment (b).

(106) When performing a therapeutic treatment (see FIGS. 18c and 18d), it is observed that compound 15 improves the clinical classification of the motor capacity of the mice compared to the control, especially between days 20-30 of the test (second week of administration of the compound). It is also striking that this improvement in the clinical classification was maintained during days 30-47, when the administration of the drug had already been interrupted. In contrast, the mice do not appear to improve in terms of the damage produced when RES is administered in therapeutic mode.

(107) It should also be highlighted that the tween-80 emulsifier appears to have a pharmacological effect per se (see controls of the preventive treatment groups, a vs. b), which would be masking, at least partially, the possible effect of compound 15 in reducing the damage in the multiple sclerosis model.

(108) It is important to note that 20 mg/kg of each compound (RES and compound 15) was administered, but given the large differences in molecular weight of the two compounds (MW compound 15=3×MW RES), the administered doses are not comparable. Thus, the dose administered (in μmol/kg) of RES≅3× dose of compound 15, so that although the two compounds show a similar efficacy in the treatment of EAE, compound 15 does so at a much lower concentration than RES and is therefore much more effective.