WO2003086414A1

WO2003086414A1 - Structural modification of resveratrol: sodium resverastatin phosphate

Info

Publication number: WO2003086414A1
Application number: PCT/US2003/011008
Authority: WO
Inventors: George R. Pettit; Matthew P. Grealish
Original assignee: Arizona Board Of Regents
Priority date: 2002-04-10
Filing date: 2003-04-10
Publication date: 2003-10-23
Also published as: US7705188B2; US20050240062A1

Abstract

Described herein are novel compounds having antineoplastic and antimicrobial activity, obtained via structural modifications of resveratrol and combretastatin A-4, methods for synthesis of these compounds, and their use in pharmaceutical composition and for use in the treatment of mammals having cancer. Examples of the novel compounds are: (Z)- and (E)- 3,4',5-trimethoxystilbene (4a, 4b); (Z)- and (E)-3,5-dimethoxy-4'-hydroxystilbene (14c, 14d); (Z)-and (E)-3-hydroxy-4',5-dimethoxystilbene (14g, 14h); (Z)-and (E)-3,5-dihydroxy-4'-methoxy-stilbene (14k, 14l); sodium resverastatin dibenzyl phosphate ((Z)-3,5-dimethoxy-4-[O-bis(benzyl)phosphoryl]-stilbene) (14m); and sodium resverastatin phosphate (14n).

Description

TITLE OF THE INVENTION Structural Modification of Resveratrol: Sodium Resverastatin Phosphate RELATED APPLICATION DATA This application is based on and claims the benefit of U.S. Provisional Patent Application No. 60/371,782 filed on April 10, 2002.

GOVERNMENT INTEREST Financial assistance for this invention was provided by the United States Government, Division of Cancer Treatment and Diagnosis, National Cancer Institute, Department of Health

and Human Services Outstanding Investigator Grant Numbers CA44344-05-12 and RO1 CA90441-01. Thus, the United States Government has certain rights in this invention.

FIELD OF THE INVENTION The present invention relates generally to antineoplastic compositions. More particularly this invention relates to derivatives of resveratrol, combretastatin A-4, and methods of synthesis thereof.

BACKGROUND OF THE INVENTION The antitumor properties of a series of structurally simple compounds derived from tropical and subtropical trees of the family Combretaceae are under ongoing investigation. The genus Combretum contains 25 species used in the traditional medical practices of Africa and India. The South African bush willow Combretum caffrum was used by the Zulu and other Southern African people as a charm to ward off enemies and in traditional medical practices.

Combretum caffrum (Eckl. Zehy.) Kuntze collected in 1973 and recollected in 1979 afforded extracts that showed activity in the astrocyte reversal (9 ASK) and murine lymphocytic leukemia screening assays of the National Cancer Institute of the U.S.A. Historically, this

1398874.1/12504.012 9 extract was the first to be successfully fractionated by means of the 9ASK system. In 1982 the isolation of the first member of the combretastatin series was disclosed. Also disclosed was its structure and later synthesis. (Pettit, G. R., et al., Isolation and Structure of Combretastatin, CAn. J. Chem. 1982, 60, 1374-1376; Pettit, G. R., et al., Synthesis of Natural (-) - combretastatin, J. Org. Chem. 1985, 50, 3404-3406.) Subsequently, a number of additional cancer cell line active constituents were isolated. These investigations eventually led to applicant's isolation, structure and synthesis of the -stilbene combretastatin A-4 (2a) and its phosphate prodrug (2b). (Pettit, G.R., et al., Isolation and Structure of the Strong Cell Growth and Tubulin Inhibitor Combretastatin A-4, Experentia, 1989, 45, 209-211; Pettit, G. R, et al., Antineoplastic agents 291. Isolation and Synthesis of Combretastatin A-4, A-5, and A-6, J. Med. Chem. 1995, 38, 1666-1672; Ndayikcngu kiyc, II., ct al., Alkoxylalcd p- phenylenevinylene Oligomers: Synthesis and Spectroscopic and Electrochemical Properties, Tetrahedron 1997, 53, 13811-13828.) The latter has been shown to selectively damage tumor neovasculature with induction of extensive blood flow shutdown in the metastatic tumor compared to normal tissues.

For example, six hours following treatment using the murine CaNT adenocarcinoma and a single i.p. injection of combretastatin A-4 prodrug (100 mg/kg), vascular function shutdown in the tumor was rapid, ineversible and extensive (8). (Ndayikengurukiye, H., et al., Alkoxylated p-phenylenevinylene Oligomers: Synthesis and Spectroscopic and Electrochemical Properties. Tetrahedron 1997, 53, 13811-13828.) In November, 1998 four Phase I human cancer trials were initiated, two in the United States and two in England. Cunent clinical trials (9) have been encouraging and Phase II human cancer clinical trials have been or will be initiated soon. (Cushman, M., et al., Synthesis and Evaluation of Analogues of (Z) - 1 - (4-methoxyphenyl) -2-

1398874.1/12504.012 7 (3, 4, 5-trimethoxyphenyl) ethane as Potential Cytotoxic and Antimitotic Agents, J. Med. Chem- 1992, 35, 2293-2306.)

The need for such compounds is both critical and ongoing. The isolation of such valuable compounds is the purpose of the present invention.

BRIEF SUMMARY OF THE INVENTION

Resveratrol (1), 3,4',5-trihydroxy-t «^-stilbene, is a phytoalexin found in grapes and certain other plants. (Orsini, F., et al., Isolation, Synthesis, and Antiplatelet Aggregation of Resveratrol 3-O-β-D-glucopyranoside and Related Compounds, J. Nat. Prod. 1997, 60, 1082- 1087; Siemann, E. H., et al., Concentration of the Phytoalexin Resveratrol in Wine, Amer. J. of Enology and Viticulture 1992, 43, 49-52; Goldberg, D., et al., A Global Survey of Trans- resveratrol Concentrations in Commercial Wines, Clin. Chem. 1995, 46, 159-1665; Adesanya, S.A., et al., Stilbene Derivatives from Cissus quandrangularis. J. Nat. Prod. 1999, 62, 1694- 1695; Arichi, H., et al., Effects of Stilbene Components of the Root of Polygonum cuspidatum Sieb. Et Zucc. On Lipid Metabolism, Chem. andPharm. Bull. 1982, 30, 1766-1770; Ingham, J., 3,5,4' - trihydroxystilbene as a Phytoalexin from Groundnuts (Arachis hypogaea). Phytochem. 1976, 15, 1791-T793.)

The compound exhibits a variety of useful biological properties including antileukemic, antibacterial, antifungal, antiplatelet aggregation, and coronary vasodilator activities. (Jeandet, P., et al., The Production of Resveratrol by Grape Berries in Different Developmental Stages, Am J. ofEnology and Viticulture 1991, 42, 41-46; Manila, E., et al., Anti-leukaemic compounds Derived from Stilbenes in Picea abies Bark, Phytochem. 1993, 33, 813-816; Kubo, M., et al., Shoyayugaku Zasshi 1981, 35, 58; Creasy, L., et al., Phytoalexin Production Potential of Grape Berries, J. of the Am. Soc. of Horticultural Science 1988, 113, 230-234; Langcake, C, et al.,

1398874.1/12504.012 Identification of Pterostilbene as a Phytoalexin from Vitis vinifera Leaves, Phytochemistry 1979, 18, 1025-1027; Langcake, R., et al., The Relationship of Resveratrol Production to Infection of Grapevine Leaves by Botrytis cenerea, Vitis 1979 18:244-253; Chung, M., et al., An Antiplatelet Principle of Veratrum formosanum, Planta Medica 1992, 58, 274-276; Inamori, Y., et al., The Ichthyotoxicity and Coronary Vasodilator Action of 3,3' - dihydroxy-α, β- diethylstilbene, Chem. Pharm. Bull. 1987, 35, 887-890.) This triphenolic stilbene (1) also has strong antioxidative and anti-inflammatory activities associated with chemopreventive properties. (Pattichis, K., et al., Inhibition of Human LDL Oxidation by Resveratrol, Lancet 1993, 1103-1108; Goldberg, D., More on Antioxidant Activity of Resveratrol in Red Wine, Clin. Chem. 1996, 42, 113-114; Pace-Asciak, C, et al., The Red Wine Phenolics Trans-resveratrol and Quercetin Block Human Platelet-Aggregation and Eicosanoid Synthesis-implications for Protection Against Coronary Heart-Disease, Clinical Chemica Acta 1995, 235, 207=219.) Resveratrol (1) has been suggested as a possible cancer chemopreventive agent based on inhibitory effects on tumor initiation, promotion, and progression. (Jang, M., et al., Cancer

Chemopreventive Activity of Resveratrol. A natural product Derived from Grapes, Science 1997, 275, 218-220; Uenobe, F., Antimutagenic Effects of Resveratrol Against Trp-P-1, Mutation Res. 1997, 373, 197-200.)

In addition to antitumor promoting activity, resveratrol (1) has displayed cancer cell growth inhibition in vitro. (Chanvitayapongs, S., et al., Amelioration of Oxidative Stress by Antioxidants and Resveratrol in PC12 cells, NeuroReport 1997, 8, 1499-1502; Mgbonyebi, O., et al., Antiproliferative Effect of Synthetic Resveratrol on Human Breast Epithelial Cells, Int. J. Oncol. 1998, 12, 865-869; Jayatilake, G., et al., Kinase Inhibition From polygonum cuspidatum 1993, 56, 1805-1810; Chun, Y., et al., Resveratrol is a Selective Human Cytochrome P450 1A1

1398874.1/12504.012 *s inhibitor, Biochem. Biophys. Res. 1999, 262, 20-24.) Importantly, resveratrol (1) has recently been shown to induce apoptosis and decrease expression of Bcl-2 in the human leukemia HL-60 cell line. (Surh, Y. J., et al., Resveratrol, an Antioxidant Present in Red Wine, Induces Apoptosis in Human Promyelocytic Leukemia (HL-60) cells, Cancer Lett. 1999, 140, 1-10.) Furthermore, the resveratrol tetramers vatdiospyridol and resveratrol oligomers recently isolated from Asian plants have shown significant inhibition of the growth of several cancer cell lines. (Seo, H., et

al., Resveratrol Tetramers from Vatica Diospyoides. J. Org. Chem. 1999, 64, 6976-6983; Ohyama, M., et al., Antitumor Agents 200. Cytotoxicity of Naturally Occurring Resveratrol Oligomers and their Acetate Derivatives, Bioorg. Med. Chem. Lett. 1999, 9, 3057-3060.) Other biological properties of resveratrol (1) include activities targeting cyclooxygenase, tyrosine kinase (PTK), and protein kinase C (PKC), as well as selective human cytochrome P450 1 Al

inhibition and microbiological transformation to resveratrol 3-O-β-D-glucoside. (Jang, M., et al.,

Cancer Chemopreventive Activity of Resveratrol. A Natural Product Derived from Grapes, Science 1997, 275, 218-220; Jayatilake, G., et al., Kinase Inhibition from Polygonum cuspidatum, J. Nat. Prod. 1993, 56, 1805-1810; Chun, Y., et al., Resveratrol is a Selective Human Cytochrome P450 1 Al Inhibitor, Biochem. Biophys. Res. 1999, 262, 20-24; Cichewicz R. H., et al., Biotransformation of resveratrol to piceid by Bacillus cereus, J. Nat. Prod. 1998, 61, 1313-1314.)

The compounds in question were isolated/synthesized in the manner generally described below. These compounds were discovered to have the properties noted.

Accordingly the prime object of the present invention was to discover pharmaceutically active derivatives of resveratrol.

1398874.1/12504.012 Another object of the present invention was to discover pharmaceutically active derivatives of combretastatin A-4.

DESCRIPTION OF THE PREFERRED EMBODIMENTS The compounds discovered were obtained in the following manner, using the following lexicon that should be familiar to one of ordinary skill in the art.

All solvents were redistilled. Hunig's base and proton sponge refer respectively to N,N,- diisopropylcthylaminc and l,8-b-_.-(dimethylamino)-naphthalcnc. Both the course and products from reactions were monitored by thin-layer chromatography using Analtech silica gel GHLF uniplates. All reactions were carried out under an inert atmosphere. Solvent extracts of aqueous solutions were dried over anhydrous sodium sulfate unless otherwise noted. Flash column chromatography was performed using silica gel (230-400 mesh ASTM).

Melting points were recorded employing an Electrothermal 9100 digital melting point apparatus and are unconected. The IR spectra were obtained using a Mattson FTIR model 2020 instrument. Low-resolution mass spectral data were collected using a Varian MAT 312 instrument (EIMS). The high-resolution FAB spectra were obtained employing a Kratos MS-50 spectrometer. All H and C-ΝMR spectra were determined using a Varian Gemini 300 MHz instrument with CDC1₃ (TMS internal reference) as solvent unless otherwise noted. The ³¹P- ΝMR spectra were measured in CDC1₃ with 85% H₃PO₄ as an external standard employing a Varian Unity 500 MHz instrument.

1398874.1/12504.012 The following schemes and structural formulae of identified compounds, set forth at the end of this application, were used. Where practical general procedures for certain classes of compounds are also set forth below.

A starting material in stilbene synthesis is 4-methoxybenzyltriphenylphosphonium bromide (3) that was obtained as follows: To a solution of 4-methoxybenzyl bromide (22.4 g) in toluene (200 ml) was added triphenylphosphine (29.2 g). The solution was heated at reflux for 12 hours under argon. The resulting precipitate was collected and recrystallized from ethanol as colorless crystals (44.0 g, 85.3%) mp 235-237°C, (lit¹⁹ mp 234°C).

The general procedure for the stilbene syntheses was as follows. To the phosphonium bromide (1-35 mmol) in anhydrous tetrahydrofuran (5-200 ml) at -78°C was added w-butyl lithium (2.44 M, 1.0 equiv.), and the resulting red solution was stined under argon for 2-4 hours. A solution of the aldehyde (1.0 equiv.) in tetrahydrofuran was added dropwise over 30 minutes and the mixture stined for 6-15 hours. The resulting cream suspension was poured into water and extracted with dichloromethane. The organic phase was washed with water and removal of the solvent in vacuo afforded a tan oil. The oil was separated by flash column chromatography (49:1 hexane:ethyl acetate). The cis stilbene eluted first as a clear oil followed by the trans isomer as a colorless sold or oil (TBDMS protected).

By following the above general procedure, 3,4',5-trimethoxystilbene (4a,b) was obtained in the following manner. Reaction of phosphonium bromide (12.5 g) (3) and 3,5- dimethoxybenzaldehyde (4.5 g) led to cis stilbene (4a) as a clear oil (3.56 g) and the trans isomer (4b) as a colorless sold (3.08 g), 91% total yield: (Z) isomer (4a) IR neat, cm^"1 v_max 3449, 2957,

2836, 1591, 1250, 1065, 640; 1H-NMR δ 6.22 (2H, dd, J= 2.4, 8.7 Hz), 6.77 (2H, dd, J= 2.4, 8.7 Hz), 6.53 (1H, d, J= 12.0 Hz), 6.45 (1H, d, J= 12.0 Hz), 6.44 (2H, d, J= 2.1 Hz), 6.32 (1H,

1398874.1/12504.012 t, J= 2.1 Hz), 3.78 (3H, s, OCH₃), 3.67 (6H, s, OCH₃ x 2); (E) isomer (4b) mp 57-58°C, (lit²⁰ mp 55-56°C).

Resveratrol (1) was obtained from a stilbene in the following manner. To stilbene 4b (3.1 g) in anhydrous dichloromethane (150 ml) at -78°C was added (dropwise) boron tribromide (1.0 M, 34.5 ml), and the resulting red solution was stined under argon for 30 minutes. The solution was poured into water and extracted with dichloromethane. The organic phase was washed with water and removal of the solvent in vacuo afforded a tan oil, which was separated by flash column chromatography (l:l/hexane: ethyl acetate) to afford a colorless sold (2.26 g, 86%): mp 260°C (lit^12c mp 260°C).

The compound 4-(tert-butyldiphenylsilyloxy)-benzaldehyde (5a) was obtained in the following manner. To a solution of 4-hydroxybenzaldehyde (3.2 g) in dimethylformamide (50 ml) was added imidazole (1.9 g, 1.1 equiv.). The solution was stined for 15 minutes, tert- butyldiphenylsilyl chloride (7.4 ml, 1.1 equiv.) was added, and the light brown solution was stined for 3 hours. The reaction mixture was poured into water and extracted with ethyl acetate. Removal of the solvent in vacuo from the organic phase provided a brown oil. The oil was

separated by flash column chromatography (1:0 — 19:1 hexane:ethyl acetate) to afford the aldehyde (5a) as a colorless solid (6.4 g, 68%):mp 103-105°C IR (neat, cm^"1) v_max 3399, 2932,

2859, 1699, 1599, 1506, 1273, 1157, 910; 1H-NMR δ 9.80 (1H, s, CHO), 7.69 (4H, m, Ar-H),

7.64 (2H, d, J= 8.7 Hz), 7.40 (6H, m, Ar-H), 6.86 (2H, d, J=8.7 Hz), 1,11 (9Hm sm C(CH₃)₃), Anal. (C₂₃H₂₄O₂Si) C, H.

The compound 4-(tert-butyldiphenylsilyloxy)-benzyl alcohol (6) was obtained in the following manner. To a solution of aldehyde 5a (4.7 g) in methanol (100 ml) at 0°C was slowly added sodium borohydride (0.59 g, 1.2 eq.). After stirring for 2 hours the reaction mixture was

1398874.1/12504.012 poured into water, solvent reduced to a minimum, extracted with ethyl acetate and the solvent

removed in vacuo to give 4.1 g of a clear oil (88%); IR (neat, cm^"1) v_max 3346, 2932, 2859, 1609,

1510, 1427, 1256, 1113, 918; 1H-NMR δ 7.72 (4H, m, Ar-H), 7.39 (6H, m, Ar-H), 7.10 (2H, d, J = 8.7 Hz), 6.76 (2H, d, J= 8.7 Hz), 4.55 (2H, s, CH₂), 1.11 (9H, s, C(CH₃)₃). Anal.

(C₂₃H₂₆O₂Si) C, H.

The following procedure was used to obtain 4-(tert-butyldiphenylsilyloxy)-benzyl bromide (7). Phosphorus tribromide (0.5 ml) was slowly added to a solution of the alcohol 6 (4.0 g) in dichloromethane (75 ml) at 0°C, and stirring was continued for 12 hours. The reaction

mixture was poured into aqueous sodium bicarbonate, extracted with dichloromethane and the

solvent removed in vacuo to afford 4.3 g of a colorless solid (89%): EIMS m/z 426 (M+, ⁸¹Br),

424 (M+ ⁷⁹Br), 390, 369, 367, 345, 289, 135: IR (neat.cm^"1) v_max 3397, 3073, 2932, 2859, 1607,

1510, 1427, 1263, 1113, 918; ^]H-NMR δ 7.72 (4H, m, Ar-H), 7.39 (6H, m, Ar-H), 7.13 (2H, d, J

= 8.4 Hz) 6.72 (2H, d, J= 8.4 Hz), 4.42 (2H, s, CH₂), 1.11 (9H, s, C(CH₃)₃). ¹³C-NMR (75.5

MHz) δ 155.6, 135.4, 132.4, 130.1, 129.9, 127.7, 119.9, 34.0, 26.5, 19.5. Anal. (C₂₃H₂₅BrOSi) C, H.

The compound 4-(tert-butyldiphenylsilyloxy)-benzyltriphenylphosphonium bromide (8) was obtained in the following manner. To a solution of bromide (7) (4.3 g) in toluene (100 ml) was added triphenylphosphine (13.2 g). After heating at 100°C for 2 hours, the reaction mixture was cooled to room temperature, and the product was collected and recrystallized from ethanol to yield 6.1 g of a colorless solid (89%) mp 233°C; FABMS m/z 607.2583 (M+-Br), IR (neat, cm^"1) v_max 3385, 3054, 2934, 2859, 2787, 1607, 1512, 1437, 1273, 1111, 924; 1H-NMR δ 7.61

(19H, m, Ar-H), 7.30 (6H, m, Ar-H), 6.76 (2H, dd, J= 2.4, 8.7 Hz) 6.51 (2H, d, J= 8.7 Hz), 5.18

(2H, s, CH₂), 1.04 (9H, s, C(CH₃)₃). ¹³C-NMR (75.5 MHz) δ 155.7, 135.3, 134.7, 134.7, 134.3,

1398874.1/12504.012 J Q 132.3, 132.3, 130.0, 129.8, 127.6, 120.2, 120.1, 118.9, 118.9, 118.1, 117.3, 30.5, 30.1, 26.5, 19.4.

Anal. (C₄ιH₄₀BrOSi) C, H.

The compound 3,5-di(tert-butyldimethylsilyloxy)-benzaldehyde (9a) was obtained in the following manner. DIEA (7.7 ml, 2 equiv.) was added to a solution of 3,5- dihydroxybenzaldehyde (3.0 g) in dimethylformamide (30 ml), and the solution was stined for 15 minutes. The silyl chloride (7.5g) was added and the light brown solution stined for 16 hours. The mixture was poured into water and extracted with dichloromethane. Removal of the solvent in vacuo yielded a brown oil that was separated by flash column chromatography (9:1 hexane:ethyl acetate) to yield the disilylether as a tan oil (7.6 g, 94%): EIMS m/z 366 (M+), 309,

267, 239, 133,73; IR (KBr, cm^"1) v_max 2957, 2861, 2805, 2710, 1705, 1385, 831; 1H-NMR δ 9.85

(1H, s, CHO), 6.95 (2H, d, J= 2.1 Hz), 6.58 (1H, t, J= 2.1 Hz), 0.99 (18H, s, C(CH₃)₃ x 2), 0.22 (12H, s, Si(CH₃)₂ x 2).

The compound identified as cw-Resveratrol (10) was obtained as follows: the Wittig reaction was performed as summarized above using 5 mmol of phosphonium salt, and the TBDPS-protected stilbene isomers were isolated as a mixture. The mixture was dissolved in tetrahydrofuran and treated with TBAF (3.0 eq), being stined for 1 hour. The product was purified by gravity column chromatography (3:2 hexane:ethyl acetate) and yielded 0.21 g of the cw-isomer as a colorless solid and 0.24 g of a mixture of isomers (95.1%): mp 172-174°C (lit^12c mp 170-174°C).

Hunig's base (10.2 ml, 2 eq) was added to a solution of 4-hydroxybenzaldehyde (6.0 g) in dimethylformamide (50 ml) to obtain 4-(tert)-butyldimethylsilyloxy-benzaldehyde (5b). The solution was stined for 15 minutes, tert-butyldimethylsilyl chloride (8.9 g) was added, and the clear light brown solution was stined for 15 hours. The reaction mixture was poured into water,

1398874.1/12504.012 \ \ extracted with dichloromethane and the solvent removed in vacuo to afford a brown oil. The oil was separated by vacuum distillation to yield aldehyde 5b as a colorless oil (8.4 g, 73%): IR

(neat, cm^"1) v_max 3385, 2932, 2859, 1699, 1599, 1508, 1273, 1155, 909; 1H-NMR δ 9.88 (IH, s,

CHO), 7.79 (2H, d, J= 8.4 Hz), 6.94 (2H, d, J= 8.4 Hz), 0.98 (9H, s, C(CH₃)₃), 0.25 (6H, s, Si(CH₃)₂). Anal. (C,₃H₂₀O₂Si) C, H.

To obtain 3,5-dimethoxybenzyltriphenylphosphonium bromide (13) 3,5 - dimethoxybenzaldehyde (10 g) in methanol was reduced with sodium borohydride, the oily product 11, (9.6 g, 93% yield) was treated (0°, 12 hours) with phosphorous tribromide (2.7 ml), and to its resulting bromide 12 (11.6 g, 89%) in toluene (200 ml) was added triphenylphosphine (13.2 g). After heating at reflux for 12 hours the mixture was cooled to room temperature. The product was collected and recrystallized from ethanol to yield 22.8 g of a colorless solid (92%)

mp 275°C, lit^12c mp 266-268°C; Η-NMR δ 7.70 (15H, m), 6.33 (2H, d, J= 2.1 Hz), 6.30 (IH, t,

J= 2.1 Hz), 5.30 (2H, d, J= 14.4 Hz), 3.53 (6H, s, OCH₃ x 2).

To obtain 4'-(tert-butyldimethylsilyloxy)-3,5-dimethoxy-stilbene (14a,b), phosphonium bromide 13 (6.9 g) in anhydrous tetrahydrofuran (40 ml) at 78°C was treated with «-butyl lithium 2.5 M, 5.6 ml) and aldehyde 5b (3.3 g) in tetrahydrofuran (10 ml) according to the general Wittig-stilbene procedure (see above). (Z)-isomer (14a): EIMS m/z 370 (M+), 355, 313,

298,157; IR (KBr, cm^"1) v_max 2932, 2857, 1591, 1508, 1262, 1155, 914; 1H-NMR δ 7.14 (2H, d,

J= 8.5 Hz), 6.70 (2H, d, J= 8.5 Hz), 6.51 (IH, d, J= 12.0 Hz), 6.43 (IH, d, J= 12.0 Hz), 6.42 (2H, d, J= 2.0 Hz), 6.31 (1 H, t, J= 2.0 Hz), 3.65 (6H, s, OCH₃ x 2), 0.96 (9H, s, C(CH₃)₃), 0.17

(6H, s Si(CH₃)₂); ¹³C-NMR (75.5 MHz) δ 160.51, 154.87, 139.40, 130.27, 130.21, 128.76,

119.77, 106.57, 99.81, 55.17, 25.65, 18.22, -4.45. Anal. (C₂₂H₃₀O₃Si) C,H. (E) isomer (14b): EIMS m/z 370 M+), 355, 313, 255 165 73; IR (KBr, cm^"1) v_max 2955, 2859, 1595, 1508, 1263,

1398874,1/12504.012 12 1154, 914, 839; 1H-NMR δ 7.40 (2H, d, J= 8.5 Hz). 7.05 (IH, d, J= 16.0 Hz), 6.92 (IH, d, J= 16.0 Hz), 6.85 (2H, d, J= 8.5 Hz), 6.66 (2H, d, J= 2.5 Hz), 6.39 (IH, t, J= 2.5 Hz), 3.84 (6H, s, OCH₃ x 2), 1.01 (9H, s, C(CH₃)₃), 0.23 (6H, s, Si(CH₃)₂); ¹³C-NMR (75.5 MHz) δ 160.9, 155.7, 139.7, 130.5 128.8, 127.7, 126.7, 120.3, 104.3, 99.6, 55.3, 25.7, 18.2, -4.4. Anal. (C₂₂H₃₀O₃Si) C, H.

(Z) and (E)-3,5-dimethoxy-4'-hydroxy-stilbene (14c and d) and General Silyloxy Deportation Procedure were performed as follows. To a solution of the silyloxy protected (Z)- stilbene (14a, 1.2 g) in anhydrous tetrahydrofuran (20 ml) was added tetrabutylammonium fluoride (1 M, 3.4 ml). The clear light yellow solution was stined for 45 minutes, poured into water, extracted with dichloromethane and the solvent removed in vacuo to provide a tan oil. The oil was separated by gravity column chromatography (4: 1 hexane-ethyl acetate) to afford cis stilbene 14c as a yellow oil (88%): IR (neat, cm¹) v_max 3385, 3005, 2940, 2837, 1591, 1512, 1456, 1152 1065, 679; 1H-NMR δ 8.01 (IH, s, OH), 7.15 (2H, d, J= 8.7 Hz), 6.71 (2H, d, J=

8.7 Hz). 6.51 (IH, d, J= 12.6 Hz), 6.43 (2H, d, J= 2.5 Hz), 6.42 (IH, d, J= 12.6 Hz), 6.31 (2H, d, J= 2.5 Hz), 3.66 (6H s, OCH₃ x 2). Anal. (Cι₆Hι₆O₃) C, C, H. (E)-3,5-dimethoxy-4'- hydroxy-stilbene (14d) was similarly prepared from stilbene 14b (0.5 g) and tetrabutylammonium fluoride (1M, 1.3 ml) in anhydrous tetrahydrofuran (10 ml) was added to yield 0.8 g of yellow oil (90%): IR (neat, cm^"1) v_max 3385, 3005, 2940, 2837, 1591, 1512, 1456, 1152, 1065,

961; Η-NMR δ 8.01 (IH, s, OH), 7.44 (2H, d, J= 8.7 Hz), 7.18 (IH, d, J= 16.5 Hz), 6.98 (IH, d, J= 16.5 Hz), 6.85 (2H, d, J= 8.7 Hz), 6.73 (2H, d, J= 2.1 Hz), 6.38 (IH, t, J= 2.1 Hz), 5.25 (IH, bs, OH), 3.81 (6H, s, OCH₃ x 2).

Unless otherwise noted the following intermediates and stilbene objectives were prepared by the preceding general methods for silyloxy protection, Wittig reaction and deprotection. The

1398874.1/12504.012 1 3 following procedure was followed to obtain 3-(tert-butyldimethylsilyloxy)-5- hydroxybenzaldehyde (9b). 3,5-dihydroxybenzaldehyde (1.1 g) in dimethylformamide (10 ml) was monosilylated using DIEA (1.9 ml, 1.4 equiv.) and the silyl chloride (1.2 g) with stirring for 3 hours. The oily product was separated by flash column chromatography (9:1 hexane:ethyl acetate) afford some disilylated product (0.7 g) and the desired monosilylated product as a colorless oil (0.8 g, 38.5%) that crystallized from ethanol: mp = 79.6-80°C; EIMS m/z 252 (M+), 195, 167, 58, 45; IR (KBr, cm^"1) v_max 3211, 2930, 2859, 1672, 1591, 1332, 841; 1H-NMR δ 9.84

(IH, s, CHO), 6.99 (IH, dd, J= 2.0, 1.0 Hz), 6.91 (IH, dd, J= 2.0, 1.0 Hz), 6.64 (IH, t, J= 2.0

Hz), 6.00 (IH, bs, OH), 0.97 (9H, s, C(CH₃)₃), 0.21 (6H, s, Si(CH₃)₂); ¹³C-NMR (75.5 MHz) δ

192.4, 157.6, 157.4, 138.3, 114.4, 114.0, 109.1, 25.6, 18.2, -4.47. Anal. (Cι₃H₂₀O₃Si) C, H

Also, 3-(tert-butyldimethylsilyloxy)-5-methoxybenzaldehyde (9c) was obtained in a like manner. To a solution of phenol 9b (0.7 g) in dichloromethane (10 ml) was added molecular sieves (4 A, 0.8 g), proton sponge (1.6 g, 2.5 eq.), and trimethyloxonium tetrafluoroborate (1.1 g, 2.5 eq), and the solution was stined for 15 hours. The solution was filtered, the sieves were rinsed with ethyl acetate and the solvent was removed from the combined filtrate in vacuo to yield a yellow oil. The oil was purified by flash column chromatography (10:1 hexane:ethyl acetate), yielding a colorless oil (0.6 g, 79%): EIMS m/z 266 (M+), 209, 181, 166, 89, 58, IR

(KBr, cm^"1) v_max 2932, 2859, 1703, 1593, 1468, 1337, 1059, 839; Η-NMR δ 9.86 (IH, s, CHO), 7.00 (IH, d, J= 2.0 Hz), 6.92 (IH, d, J= 2.0 Hz), 6.63 (IH, t, J= 2.0 Hz), 3.81 (3H, s, OCH₃),

0.97 (9H, s, C(CH₃)₃), 0.21 , (6H, Si(CH₃)₂); ¹³C-NMR (75.5 MHz) δ 191.8, 161.2, 157.3,

138.4, 114.5, 113.0, 106.6, 55.5, 25.6, 18.2, -4.5. Anal. (C_HH₂₂O₃Si) C,H.

In addition, 3-(tert-butyldimethylsilyloxy)-5,4'-dimethoxy-stilbene (14e and 14f) was obtained in a similar fashion. Reacting phosphonium bromide 8 (1.71 g) with aldehyde 9c (1.0

1398874.1/12504.012 \_\ g) led to stilbenes 14e and 14f (0.75 g, 55% total yield). (Z)-isomer (14e): EIMS m/z 370 (M+),

313, 298, 156, 89; IR (KBr, cm^"1) v_max 2955, 2859, 1588, 1510, 1433, 1252, 1159, 1034, 839,

679; 1H-NMR δ 7.21 (2H, d, J= 9.0 Hz), 6.77 (2H, d, J= 9.0 Hz), 6.52 (IH, d, J= 12.0 Hz),

6.45 (IH, s) 6.43 (IH d, J= 12.0 Hz), 6.36 (IH, d, J= 2.1 Hz), 6.27 (IH, t, J= 2.1 Hz), 3.78 (3H, s, OCH₃), 3.67 (3H, s, OCH₃), 0.95 (9H, s, C(CH₃)₃), 0.11 (6H, s, Si(CH₃)₂); ¹³C-NMR

(75.5 MHz) (75.5 MHz) δ 160.6, 158.8, 156.7, 139.4, 130.3, 128.8, 127.5, 126.3, 113.6,110.9,

105.4, 55.2, 25.3, 14.1, -4.5. Anal. (C₂₂H₃₀O₃Si) C,H. (E)-isomer (14f): EIMS m/z 370 (M+),

313, 298, 156, 89; IR (KBr, cm^"1) v_max 2955, 2859, 1588, 1510, 1433, 1252, 1159, 1034, 941,

839; 1H-NMR δ 7.45 (2H, d, J= 8.7 Hz), 7.01 (IH, d, J= 15.9 Hz), 6.90 (2H, d, J= 8.7 Hz),

6.87 (IH, d, J= 15.9 Hz), 6.66 (IH, s), 6.58 (IH, s), 6.31 (IH, t, J= 2.1 Hz), 3.83 (3H, s, OCH₃),

3.81 (3H, s, OCH₃), 1.00 (9H, s, C(CH₃)₃), 0.23 (6H, s, Si(CH₃)₂); ¹³C-NMR (75.5 MHz) δ 160.8, 159.4, 156.9, 139.6, 130.0, 128.5, 127.8, 126.6, 114.1, 110.9, 104.8, 55.3, 25.7, 14.1, - 4.4. Anal. (C₂₂H₃₀O₃Si) C, H.

A similar process was followed to obtain 3-hydroxy-4',5-dimethoxystilbene (14g and 14h). The preceding stilbene (0.75 g) isomeric mixture was deprotected and the products separated by gravity column chromatography (9:1 hexane:ethyl acetate). As usual the cis- stilbene (0.25 g) eluted first followed by the trans-isomer (0.26 g, 99% total yield): (_Z)-isomer

(14g): EIMS m/z 256 (M+), 225, 181, 152, 115; IR (KBr, cm^"1) v_max 3407, 3005, 2938, 2837,

1607, 1511, 1456, 1300, 1254, 1154 1057, ' H-NMR δ 7.20 (2H, d, J= 9.0 Hz), 6.77 (2H, d, J=

9.0 Hz), 6.52 (IH, d, J= 12.0 Hz), 6.42 (IH, s), 6.40 (IH d, J= 12.0 Hz), 6.33 (IH, d, J= 2.1 Hz), 6.27 (IH, t, J= 2.1 Hz), 3.79 (3H, s, OCH₃), 3.67 (3H, s, OCH₃). Anal. (Cι₆Hι₆O₃) C,H.

(EHsomer (14h): EIMS m/z 256 (M+), 225, 181, 152, 115; IR (KBr, cm^"1) v_max 3405, 2936,

2837, 1593, 1510, 1456, 1252, 1150,1057; and 1H-NMR δ 7.44 (2H, d, J= 8.7 Hz), 7.02 (IH, d,

1398874.1/12504.012 15 J= 15.9 Hz), 6.90 (2H, d, J= 8.7 Hz), 6.86 (IH, d, J= 15.9 Hz), 6.63 (1 H, t, J= 2.1 Hz), 6.58 (1 H, t, J= 2.1 Hz), 6.31 (1 H, t, J= 2.1 Hz), 3.83 (3H, s, OCH₃) 3.82 (3H, s, OCH₃).

In a like manner 3,5-di(tert-butyldimethylsilyloxy)-4'-methoxy-stilbene (14i and 14j) was obtained. Intermediate 9a and 3 served as starting material for preparing stilbene 14i (1.73 g)

and 14j (0.19 g). (Z)-isomer (14i); EIMS m/z 470 (M+), 455, 413, 147, 73; IR (KBr, cm^"1) v_max

2955, 2859, 1582, 1510, 1437, 1331, 1254, 1165, 1031, 678; 1H-NMR δ 7.17 (2H, d, J= 8.1

Hz), 6.75 (2H, d, J= 8.1 Hz), 6.49 (1 H, d, J= 12.0 Hz), 6.39 (IH, d, J= 12.0 Hz), 6.35 (2H, d, J = 2.4 Hz), 6.19 (IH, t, J= 2.4 Hz), 3.77 (3H, s, OCH₃), 0.93 (18H, s, C(CH₃)₃ x 2), 0.10 (12H, s, Si(CH₃)₂ x 2). Anal. (C₂₅H₄₂O₃Si₂) C.H. (E)-isomer (14j); EIMS m/z 470 (M+), 455, 413, 147, 73; IR (KBr, cm^"1) v_max 2955, 2859, 1582, 1510, 1437, 1331, 1254, 1165, 1031, 980; Η-NMR δ 7.44 (2H, d, -7= 8.1 Hz), 6.97 (IH, d, J= 16.2 Hz), 6.89 (2H, d, J= 8.1 Hz), 6.83 (1 H, d, J= 16.2 Hz), 6.59 (2H, d, J= 2.1 Hz), 6.24 (114, t, J= 2.1 Hz), 3.83 (3H, s, OCH₃), 1.00 (18H, s, C(CH₃)₃ x 2), 0.22 (12H, s, Si(CH₃)₃ x 2).

The preceding silyloxy protected stilbene isomers (14i and j) were deprotected to yield 3,5-dihydroxy-4'-methoxy-stilbene (14k and 1) 0.60 g and 0.05 g respectively. (Z)-isomer (14k);

EIMS m/z 242 (M+), 226, 211, 194, 181, 152, 137, IR (KBr, cm^"1) v_max 3356, 3009, 2971, 2837,

1605, 1510, 1254, 1154, 1005, 677; Η-NMR δ 7.20 (2H, d, J= 8.7), 6.77 (2H, d, J= 8.7 Hz),

6.50 (IH, d, J= 12.0 Hz) 6.36 (IH, d, J= 12.0 Hz), 6.32 (2H, d, J= 2.1 Hz), 6.22 (IH, t, J= 2.1 Hz), 4.89 (2H, bs, OH x2), 3.77 (3H, s, OCH₃). Anal. (Cι₅Hι₄O₃) C,H. E-Isomer (141); EIMS

m/z 242, (M+), 226, 211, 194, 181, 152,137; IR (KBr, cm^"1) v_max 3356, 3009, 2971, 2837, 1605,

1510, 1254, 1154, 1005, 974; Η-NMR δ 7.43 (2H, d, J= 8.1 Hz), 7.01, (IH, d, J = 15.9 Hz),

6.90 (2H, d, J= 8.1 Hz), 6.83 (IH, d, J= 15.9 Hz), 6.56 (2H, d, J= 2.4 Hz), 6.25 (IH, t, J= 2.4 Hz), 4.70 (2H, bs, OH x 2), 3.83 (3H, s, OCH₃).

1398874.1/12504.012 tfj Then (Z)-3,5-dimethoxy-4 -[O-bis(benzyl)phosphoryl]-stilbene (14m) was obtained as follows. A mixture of phenol 14c (3.9 g) and N,N-dimcthyla inopyridinc (0.2 g) in anhydrous acetonitrile (30 ml) was cooled to -10°C, and carbon tetrachloride (7.3 ml, 5 equiv.) and DIEA (5.5 ml, 2.1 equiv.) were added. The mixture was stined at -10°C for 30 minutes under argon, dibenzylphosphite (5.0 ml, 1.5 equiv.) was added, .and the solution was stined for 12 hours and then poured into 0.5 M monobasic potassium phosphate. The mixture was extracted with ethyl acetate and removal of solvent in vacuo from the organic phase yielded a tan oil. This was subjected to column chromatography (4:1 hexane:ethyl acetate), and the phosphate ester was recovered as a tan oil (6.6 g, 85%): EIMS m/z 516 (M+), 425, 334, 319, 255, 227, 91; IR (KBr, cm^"1) v_max 3443, 3007, 2959, 2837, 1591, 1505, 1456, 1289, 1208, 1155, 1015, 953: 1H-ΝMR δ 7.32 (10H, m, AR-H), 7.19 (2H d, J= 8.4 Hz), 7.01 (2H, d, J= 8.4 Hz), 6.52 (2H, s, H,_a>,._a), 6.36 (2H, d, J= 2.0 Hz), 6.31 (IH, t, J= 2.0 Hz), 5.11 (2H, s, Bn), 5.08 (2H, s, Bn), 3.62 (6H, s, OCH₃ x 2); ¹³C-NMR (75.5 MHz) δ 160.6, 149.5, 149.5, 138.8,135.4, 135.4, 134.2, 130.4, 130.3, 129.4, 128.6, 128.6, 128.0, 127.0, 119.8, 119.7, 106.6, 99.8, 70.0, 69.9, 55.2. Anal. (C₃₀H₂₉O₆P) C, H, P.

Sodium Resverastatin Phosphate (14n) was obtained as described below. To a solution of the dibenzyl phosphate (14m, 2.62 g) in anhydrous dichloromethane (15 ml) at 0°C was added bromotrimethylsilane (1.40 ml, 2.1 equiv.), and the mixture was stined for 2 hours. Water (10 ml) was added, the solution was stined for 1 hour and then washed with ethyl acetate, and the aqueous phase was freeze-dried to a white solid. To a solution of the solid in ethanol (30 ml) was added sodium methoxide (0.57 g), and the suspension was stined for 12 hours. Solvent was removed in vacuo and the resulting tan oil was dissolved in water. The solution was washed with ethyl acetate and then freeze-dried to afford 1.88 g of colorless solid (98%): HRFAB m/z;

1398874.1/12504.012 17 IR (KBr, cm^"1) v_max 3385, 2999, 2938, 2834, 1601, 1508, 1366, 1155, 1063, 683; 1H-NMR δ

6.93 (2H, d, J= 8.4 Hz), 6.85 (2H, d, J= 8.4 Hz), 6.29 (1 H, d, J= 12.4 Hz), 6.17 (2H, s, H_2>6),

6.15 (IH, d, J= 12.4 Hz), 6.09 (IH, s, H4),3.34 (6H, s, OCH₃ x 2); ¹³C-NMR (75.5 MHz) δ

171.1, 160.1, 139.9, 131.8, 130.8, 130.0, 129.0, 120.3, 107.3, 99.7, 55.6.

The following general procedure was used for benzhydrol formation. To the bromide (1- 10 mmol) in anhydrous tetrahydrofuran (5-35 ml) at -78°C was added n-butyllithium (2.5 M, 1.1 equiv.) and the resulting solution stined under argon for 15 minutes. A solution of aldehyde (1.0 equiv.) in tetrahydrofuran was added dropwise over 30 minutes and the mixture stined for 6 hours. The solution was poured into water and extracted with ethyl acetate. The organic phase was washed with water and removal of the solvent in vacuo afforded an oil that was purified by flash column chromatography (9:1 hexane-ethyl acetate).

Reacting 4-bromoanisole (2.3 g) and 3,5-dimethoxybenzaldehyde (2.1 g) led to 15a (2.7 g, 80.9%) as a colorless oil: EIMS m/z 274 (M+) 257, 243, 227, 165, 139, 135, 109, 77; IR

(KBr, cm^"1) v_max 3451, 3001, 2940, 2837, 1597, 1248, 1172, 1034; 1H-NMR δ 7.28 (2H, d, J=

8.8 Hz), 6.85 (2H, d, J= 8.8 Hz), 6.53 (2H, d, J= 2.0 Hz), 6.35 (IH, t, J= 2.0 Hz), 5.72 (IH, d, J

= 2.8 Hz), 3.78 (3H, s, OCH₃), 3.76 (3H, s, OCH₃); ¹³C-NMR (75.5 MHz) δ 160.9, 159.1, 146.5,

135.9, 127.9, 113.9, 104.4, 99.3, 75.8, 55.3, 55.3. Anal. (C_.6Hι₈O₄) C, H.

The following general procedure was followed for benzophenone formation. To a solution of the benzhydrol (1-10 mmol) in dichloromethane (5-35 ml) was added pyridinium dichromate (2.0 equiv.) and molecular sieves (4 A activated powder, same weight as PDC), and the resulting suspension was stined under argon for 24 hours. The reaction mixture was filtered through celite and the solvent removed in vacuo to yield a brown oil. The oil was subjected to

1398874.1/12504.012 l g gravity column chromatography (4:1 hexane-ethyl acetate) to yield the desired product in 75-

90% yield.

Further, 3,4',5-trimethoxybenzophenone (16a) was obtained. Alcohol 15a (1.1 g) provided a solid (0.8 g, 77.7%) that recrystallized from methanol in colorless needles: m.p. = 90.3-91.6°C (lit.²¹ m.p. = 97-98°C); EIMS m/z 272 (M⁺), 257, 241, 229, 165, 135, 92, 77; IR (KBr.cm^"1) v_max 3071, 2967, 2841, 1645, 1588, 1263, 1065; 1H-NMR δ 7.84 (2H, d, J= 9.0 Hz),

6.95 (2H, d, J= 9.0 Hz), 6.87 (2H, d, J= 2.0 Hz), 6.65 (IH, t, J= 2.0 Hz), 3.88 (3H, s, OCH₃),

3.82 (3H, s, OCH₃); ¹³C-NMR (75.5 MHz) δ 195.2, 163.3, 160.5, 140.2, 132.6, 130.1, 113.6,

107.6, 104.2, 55.6, Anal. (Cι₆Hι₆O₄) C, H.

Then l-bromo-4-O-(/ert-butyldimethylsilyloxy)benzene (17) was obtained. First, 4-

bromophenol (4.15 g) was dissolved in anhydrous dichloromethane (40 ml), then imidazole (1.63 g) and tert-butyldimethylsilyl chloride (3.61 g) were added. The cream suspension was stined under argon for 12 hours and the reaction was terminated with addition of water. The mixture was extracted with ethyl acetate. The organic phase was washed with water and the solvent was removed in vacuo to afford a yellow oil that was subjected to flash column chromatography (9:1 hexane:ethyl acetate) to yield a colorless oil (87%, 6.0g): EIMS m/z 274 (M⁺), 257, 243, 227,

165, 139, 135, 109, 77; IR (KBr, cm^"1) v_max 2957, 2859, 1588, 1487, 1458, 910, 839; 1H-NMR δ

7.32 (2H, d, J= 8.7 Hz), 6.71 (2H, d, J=8.7 Hz), 0.97 (9H, s, C(CH₃)₃), 0.18 (6H, s, Si(CH₃)₂). Anal (Cι₂H_]9BrOSi) C, H.

Protected bromophenol 17 (3.2 g) and 3,5-dimethoxybenzaldehyde (1.8 g) afforded 15b as a colorless oil (3.4 g, 83%): EIMS m/z 374 (M⁺), 317, 167, 151, 139; IR (KBr, cm^"1 v_max

3420, 2957, 2859, 1607, 1260, 1155, 1063, 839; 1H-NMR δ 7.21 (2H, d, J= 8.4 Hz), 6.80 (2H, d, J= 8.4 Hz), 6.53 (2H, d, J= 2.0 Hz), 6.36 (IH, t, J= 2.0 Hz), 5.67 (IH, s), 3.75 (6H, s, OCH₃

1398874.1/12504.012 19 x 2), 0.99 (9H, s, C(CH₃)₃), 0.20 (6H, s, Si(CH₃)₂); ¹³C-NMR (75.5 MHz) δ 160.7, 155.0, 146.5,

136.4, 127.8, 119.9, 104.4, 99.2, 75.7, 55.2, 25.6, 18.1, -4.5. Anal. (C₂ιH₃₀O₄Si) C, H.

Alcohol 15b (0.12g) was deprotected with TBAF as for the stilbenes above to provide 4'- Hydroxy-3,5-dimethoxybenzhydrol 15c as a colorless oil (0.03 g, 42%): EIMS m/z 260 (M⁺),

243, 165, 139, 121, 95; IR (KBr, cm^"1) v_max 3362, 2932, 2859, 1599, 1256, 1155, 1067; 1H-NMR

δ 7.16 (2H, d, J= 8.4 Hz), 6.72 (2H, d, J= 8.4 Hz), 6.52 (2H, d, J= 2.4 Hz), 6.34 (IH, t, J= 2.4

Hz), 5.66 (IH, s), 3.73 (6H, s, OCH₃ x 2); ¹³C-NMR (75.5 MHz) δ 160.6, 155.4, 146.3, 135.3, 128.0, 115.3, 104.4, 99.2, 75.8, 55.3. Anal. (Cι₅H,₆O₄) C, H.

Alcohol 15b (0.58 g) led to 4'-(ter/-butyldimethylsilyloxy)-3,5-dimethoxybenzophenone 16b as a colorless oil (0.46 g, 80%): EIMS m/z 372 (M⁺), 315, 165, 157, 137, 28; IR (KBr, cm^"1)

v_max 2957, 2859, 1657, 1260, 1067, 910, 841; Η-NMR δ 7.78 (2H, d, J= 9.2 Hz), 6.89 (2H, d, J

= 9.2 Hz), 6.87 (2H, t, J= 2.0 Hz), 6.63 (IH, t, J= 2.0 Hz), 3.80 (6H, s, OCH₃ x 2), 0.98 (9H, s,

C(CH₃)₃), 0.23 (6H, s, Si(CH₃)₂); ¹³C-NMR (75.5 MHz) δ 195.0, 160.3, 159.8, 140.0, 132.3,

130.5, 119.6, 107.5, 104.1, 55.5, 25.6, 18.3, -4.3. Anal. (C₂.H₂₈O₄Si) C;H.

The general procedure for benzophenone deprotection with TBAF is set forth below. To a solution of the protected phenol (0.3-3.5 mmol) in anhydrous tetrahydrofuran (5-25 ml) was added tetrabutylammonium fluoride (1 M, 1.0 equiv. per TBDMS), and the pale yellow solution was stined for 45 minutes. The mixture was poured into water and extracted with ethyl acetate. Removal of the solvent in vacuo from the organic phase afforded a tan oil that was subjected to gravity column chromatography (9:1 hexane:ethyl acetate) to afford the product (70-93% yield).

Protected benzophenone 16b (0.71 g) led to 16c as a white solid (0.35 g, 72%): EIMS

m/z 258 (M⁺), 243, 227, 199, 165, 121, 45; IR (KBr, cm^"1) v_max 3424, 2940, 1640, 1591, 1454,

1206, 1157, 1065; 1H-NMR δ 7.68 (2H, d, J= 8.8 Hz), 6.80 (2H, d, J= 8.8 Hz), 6.75 (2H, d, J=

1398874.1/12504.012 20 2.0 Hz), 6.53 (IH, t, J= 2.0 Hz), 3.70 (6H, s, OCH₃ x 2);¹³C-NMR (75.5 MHz) δ 196.0, 162.3,

160.4, 140.2,133.1, 128.6, 115.6, 107.5, 104.2, 55.5. Anal. (Cι₅H,₄O₄) C, H.

Combining 4-bromoanisole (0.43 g) and aldehyde 9a (0.76 g) provided 3,5-di-(tert- butyldimethylsilyloxy)-4'-methoxybenzhydrol 15d as a faint yellow oil (0.80 g, 82%): EIMS

m/z 474 (M⁺), 459, 417, 361, 343, 73; IR (KBr, cm^"1) v_max 3420, 2932, 2859, 2361, 1591, 1451,

1252, 1163, 1026, 833; 1H-NMR δ 7.29 (2H, d, J= 9.04 Hz), 6.89 (2H, d, J= 9.0 Hz), 6.52 (2H,

d, J= 2.0 Hz), 6.28 (IH, t, J= 2.0 Hz), 5.70 (IH, s), 3.83 (3H, s, OCH₃), 1.00 (18H, s, C(CH₃)₃ x

2), 0.20 (12H s, Si(CH₃)₂ x 2); ¹³C-NMR (75.5 MHz) δ 159.0, 156.5, 146.1, 136.0, 127.9, 113.7,

111.5, 111.0, 75.5, 55.3, 25.7, 18.2, -4.4. Anal. (C₂₆H_,₂O₄Si₂) C, H.

Alcohol 15d (0.72 g) provided 3,5-di-(tert-butyldimethylsilyloxy)-4'- methoxybenzophenone 16d as a colorless oil (0.64 g, 90%): EIMS m/z 472 (M⁺), 457, 415, 359,

135, 73, IR (KBr, cm^"1) v_max 2957, 2932, 2859, 1657, 1586, 1437, 1339, 1254, 1169, 831; Η-

NMR δ 7.82 (2H, d, J= 9.0 Hz), 6.94 (2H, d, J= 9.0 Hz), 6.80 (2H, d, J= 1.5 Hz), 6.52 (IH, t, J

= 1.5 Hz), 3.88 (3H, s, OCH₃), 0.97 (18H, s, C(CH₃)₃ x 2), 0.19 (12H, s, Si(CH₃)₂ x 2); ¹³C-

NMR (75.5 MHz) δ 195.0, 163.2, 156.3, 140.1, 132.5, 130.2, 115.6, 114.7, 113.5, 55.5, 25.7,

18.2, -4.4. Anal. (C₂₆H₄₀O₄Si₂) C, H.

Protected benzophenone 16d (0.54 g) led to 16e as a white solid (0.36 g, 93%): EIMS

m/z 244 (M⁺), 227, 135, 107, 92; IR (KBr, cm^"1) v_max 3300, 2972, 2841, 2361, 1692, 1591, 1451,

1350, 1263,1171,1030; 1H-NMR δ 8.60 (2H, bs, OH x 2), 7.80 (2H, d, J= 9.0 Hz), 7.06 (2H, d, J

= 9.0 Hz), 6.69 (2H, d, J= 2.5 Hz), 6.59 (IH, t, J= 2.5 Hz), 3.91 (3H, s, OCH₃); ¹³C-NMR (75.5

MHz) δ 195.4, 164.6, 159.7, 141.8, 133.4, 131.5, 114.8, 109.3, 107.2, 56.4. Anal. (C_!4H_I2O ) C, H.

1398874.1/12504.012 21 Protected bromophenol 17 (0.49 g) and aldehyde 9a (0.62 g) afforded 15e as a faint

yellow oil (0.76 g, 78%): EIMS m/z 574 (M⁺), 559, 517, 461, 443, 73; IR (KBr, cm^"1) v_max 3420,

2932, 2859, 2361, 1591, 1451, 1252, 1163, 1026, 833; 1H-NMR δ 7.18 (2H, d, J= 8.5 Hz), 6.79

(2H, d, J= 8.5 Hz), 6.48 (2H, d, J= 2.0 Hz), 6.25 (IH, t, J= 2.0 Hz), 5.63 (IH, s, CH), 0.99 (9H, s, C(CH₃)₃), 0.97 (18H, s, C(CH₃)₃ x 2), 0.19 (6H, s, Si(CH₃)₂ x 2), 0.17 (12H, s, Si(CH₃)₂ x2);

¹³C-NMR (75.5 MHz) δ 156.4, 155.0, 146.2, 136.7, 127.9, 119.9, 111.6, 111.0, 75.4, 25.7, 18.2, -

4.4, -4.5. Anal. (C₃ιH₅₄O₄Si₃) C, H.

Alcohol 15e (0.50 g) led to 16f as a colorless oil (0.44 g, 89%): EIMS m/z 572 (M⁺),

515, 459, 323, 193, 73; IR (KBr, cm^"1) v_max 2932, 2861, 1661, 1589, 1437, 1339, 1256, 1167,

831; lH-NMR δ 7.74 (2H, d, J= 8.5 Hz), 6.88 (2H, d, J= 8.5 Hz), 6.80 (2H, d, J= 2.5 Hz), 6.52

(IH, t, J= 2.5 Hz), 0.99 (9H, s, C(CH₃)₃), 0.97 (18H, s, C(CH₃)₃ x 2), 0.24 (6H, s, Si(CH₃)₂),

0.19 (12H, s, Si(CH₃)₂ x 2); ¹³C-NMR (75.5 MHz) δ 195.2, 159.9, 156.3, 140.0, 132.4, 130.7,

119.7, 115.7, 114.7, 25.7, 25.6, 18.3, 18.2, -4.4. Anal. (C₃ιH₅₂O₄Si₃) C,H.

Protected benzophenone 16f (0.42 g) led to 3,4',5-trihydroxybenzophenone 16g as a

white solid (0.12 g, 73%): EIMS m/z 230 (M⁺, 137, 121, 93, 65, 28; IR (KBr, cm^"1) v_max 3300,

2974, 1692, 1593, 1346, 1260, 1167; 1H-NMR δ 7.73 (2H, d, J= 8.5 Hz), 6.95 (2H, d,_.J= 8.5

Hz), 6.68 (2H, d, J= 2.0 Hz), 6.57 (IH, t, J= 2.0 Hz); ¹³C-NMR (75.5 MHz) δ 195.0, 162.4,

159.2, 141.6, 133.3, 130.2, 115.9, 108.8, 106.7. Anal. (Cι₃Hι₀O₄) C, H.

Protected bromophenol 17 (1.07 g) and aldehyde 9c (0.99 g) provided 15f as a faint

yellow oil (1.19 g, 67%): EIMS m/z 474 (M⁺), 459, 417, 361, 343, 73; IR (KBr, cm^"1) v_max 3397,

2932, 2859, 1595, 1508, 1256, 1159, 839; Η-NMR δ 7.19 (2H, d, J= 8.4 Hz), 6.78 (2H, d, J= 8.4 Hz), 6.55 (IH, s), 6.42 (IH, s), 6.29 (IH, s), 5.67 (IH, s, CH), 3.74 (3H, s, OCH₃), 0.96 (9H, s, C(CH₃)₃), 0.95 (9H, s, C(CH₃)₃), 0.17 (6H, s, Si(CH₃)₂), 0.15 (6H, s, Si(CH₃)₂); ¹³C-NMR

1398874.1/12504.012 22 (75.5 MHz) δ 160.5, 156.6, 155.0, 146.2, 136.4, 127.8, 119.9, 110.8, 105.1, 105.0, 75.7, 55.3,

25.7,18.3), -4.3. Anal. (C₂₆H₄₂O Si₂) C, H.

Alcohol 15f (0.91 g) led to 16h as a colorless oil (0.71 g, 78%): EIMS m/z 472 (M⁺),

415, 223, 193, 179, 73; IR (KBr, cm^"1) v_max 2932, 2859, 1659, 1595, 1507, 1258, 1163, 839; 1H-

NMR δ 7.76 (2H, d, J= 8.8 Hz), 6.90 (IH, t, J= 2.4 Hz), 6.88 (2H, d, J= 8.8 Hz), 6.77 (IH, t, J

= 2.4 Hz), 6.58 (IH, t, J= 2.4 Hz), 3.80 (3H, s, OCH₃), 0.99 (9H, s, C(CH₃)₃), 0.97 (9H, s,

C(CH₃)₃), 0.24 (6H, s, Si(CH₃)₂), 0.20 (6H, s, Si(CH₃)₂); ¹³C-NMR (75.5 MHz) δ 195.0, 160.3,

159.8, 156.3, 140.0, 132.3, 130.6, 119.6, 114.2, 110.1, 107.6, 55.5, 25.7, 25.7, 18.3, 18.3, -4.3.

Anal. (C₂₆H₄oO₄Si₂) C, H.

Protected benzophenone 16h (0.61 g) led to 3,4'-dihydroxy-5-methoxybenzophenone 16i as a white solid (0.26 g, 82%): mp 179-180°C; EIMS m/z 244 (M⁺), 229, 213, 151, 121, 93; IR (KBr, cm^"1) v_max 3333, 2961, 2841, 1690, 1589, 1435, 1346, 1165, 1059, 849; Η-NMR δ 9.21 IH bs, OH), 8.66 (IH, bs, OH), 7.75 (2H, d, J= 9.0 Hz), 6.96 (2H, d, J= 9.0 Hz), 6.77 (2H, t, J = 2.0 Hz), 6.74 (IH, t, J= 2.0 Hz), 6.63 (IH, t, J= 2.0 Hz), 3.80 (3H, s, OCH₃); ¹³C-NMR (75.5 MHz) δ 195.0, 162.6, 161.8, 159.2, 141.6, 133.4, 130.1, 116.0, 109.9, 107.0, 105.50, 55.8. Anal.

(Cι₄H]₂O₄) C, H.

Combining 4-bromoanisole (0.59 g) and aldehyde 9c (0.82 g) provided compound 15g as a faint yellow oil (0,59 g, 52%): EIMS m/z 374 (M⁺), 359, 317, 299, 243, 75; IR (KBr, cm^"1)

v_max 3418, 2932, 2859, 1595, 1462, 1250, 1157, 1036, 837; 1H-NMR δ 7.30 (2H, d, J= 8.4 Hz), 6.89 (2H, d, J= 8.4 Hz), 6.58 (IH, s), 6.49 (IH, s), 6.32 (IH, s), 5.72 (IH, s, CH), 3.82 (3H, s, OCH₃), 3.77 (3H, s OCH₃), 0.99 (9H, s, C(CH₃)₃), 0.20 (6H, s, Si(CH₃)₂); ¹³C-NMR (75.5 MHz)

δ 160.5, 158.9, 156.6, 146.2, 135.9, 127.8, 113.8, 110.7, 105.1, 104.9, 75.6, 55.3, 25.8, 18.3, - 4.3. Anal. (C₂ιH₃₀O₄Si) C,H.

1398874.1/12504.012 23 Alcohol 15g (0.54 g) led to colorless oil 3'-(tert-butyldimethylsilyloxy)-4',5- dimethoxybenzophenone 16j (0.41 g, 76%): EIMS m/z 372 (M⁺), 315, 272, 135; IR (KBr, cm^"1) v_max 2932, 2859, 1657, 1593, 1454, 1429, 1339, 1254, 1161, 1034, 839; Η-NMR δ 7.83 (2H, d, J= 8.8 Hz), 6.95 (2H, d, J= 8.8 Hz), 6.89 (IH, t, J= 1.2 Hz), 6.77 (IH, t, J= 1.6 Hz), 6.58 (IH, t, J= 2.2 Hz), 3.88 (3H, s, OCH₃), 3.80 (3H, s, OCH₃), 0.97 (9H, s, C(CH₃)₃), 0.20 (6H, s,

Si(CH₃)₂); ¹³C-NMR (75.5 MHz) δ 194.9, 163.1, 160.4, 156.3, 140.1, 132.5, 130.0, 114.2, 113.4,

110.1, 107.6, 55.6, 55.5, 25.7, 18.1, -4.3. Anal. (C₂ιH₂₈O₄Si) C,H.

Protected benzophenone 16j (0.37 g) led to 4',5-dimethoxy-3-hydroxybenzophenone 16k

as a colorless solid (0.18 g, 70%): EIMS m/z 258 (M⁺), 227, 135, 92, 77; IR (KBr, cm^"1) v_max

3354, 3005, 2938, 2841, 1692, 1636, 1593, 1433, 1346, 1256, 1171, 1030; Η-NMR δ 8.67 (IH,

bs, OH), 7.80 (2H, dd, J= 6.8, 2.0 Hz), 7.05 (2H, dd, J= 6.8, 2.0 Hz), 6.77 (IH, dd, J= 2.0, 1.6 Hz), 6.74 (IH, dd, J= 2.4, 1.2 Hz), 6.64 (IH, t, J= 2.4 Hz), 3.90 (3H, s, OCH₃), 3.80 (3H, s,

OCH₃); ¹³C-NMR (75.5 MHz) δ 194.6, 164.0, 161.5, 159.0, 141.1, 132.8, 130.8, 114.3, 109.7,

106.9, 105.4, 55.9, 55.7. Anal. (C₁₅Hι₄O₄) C, H.

The following results lead to some observations. Because of the interesting biological properties of resveratrol (1), combined with the remarkable in vivo anticancer activity of combretastatin A-4 (2a) and its sodium phosphate prodrug (2b), a series of resveratrol structural modifications were investigated as an extension of applicant's combretastatin and phenstatin (2c) SAR research. (Ndayikengurukiye, H., et al., Alkoxylated p-phenylenevinylene Oligomers: Synthesis and Spectroscopic and Electrochemical Properties, Tetrahedron 1997, 53, 13811- 13828.) Suitable application of the experimental procedures already developed for synthesis of combretastatin A-4 and its prodrug derivatives was extended to obtaining the cis and trans stilbenes as well as the benzophenones. (Pettit, G. R., et al., Antineoplastic agents 291. Isolation

1398874.1/12504.012 24 and synthesis of Combretastatin A04, A-5, and A-6, J. Med. Chem. 1995, 38, 1666-1672; Pettit, G. R., et al., Antineoplastic agents 443. Synthesis of the Cancer Cell Growth Inhibitor Hydocyphenstatin and its Sodium Diphosphate Prodrug, J. Med. Chem. 2000, 43, 2731-2727.)

The cytotoxicity data from the resveratrol stilbenes was determined. The cΛs-isomer of resveratrol (lb) exhibited slightly less inhibitory effects on the cancer cell lines tested than did the tranj-isomer. The trimethoxy stilbenes 4a,b are between 10-100 fold more active against tumor cell lines than the parent compound resveratrol (1), with compound 4a (the α^'s-stilbene) far more active than 4b.

Demethylation at any position yielded much less cytotoxic compounds than 4a, but the cw-stilbenes 14c, 14g and 14k all retained anticancer activity comparable to that of resveratrol. The conesponding traws-isomers 14d, 14h and 141 were all slightly less active than their cis counterparts.

Previous work by applicant had found that phenstatin (2c) retained most of the cytotoxic properties of combretastatin A-4 (2a); this suggested that the properties of compound 16a, as well as additional benzophenones might possibly be worthy of further investigation. (Pettit, G. R., et al., Antineoplastic Agents. 379. Synthesis of Phenstatin Phosphate, J. Med. Chem. 1998, 41, 1688-1695.) While the cytotoxic properties of most of these compounds were quite similar to those of the stilbene derivatives of resveratrol the trimethoxy derivative was again the most potent of the series. However, it was 10- 100-fold less active than compound 4a, the analogous cw-stilbene.

General conclusions arising from the structure-activity relationship study of resveratrol based on cytotoxic effects on P-388 and human tumor cell lines can be summarized as: 3,4',5- OCH₃»resveratrol,4'-OH,prodrug,3-OH>3,5-OH,3,4'-OH»3,4',5-OH. The trimethoxy

1398874.1/12504.012 25 derivatives (4a,b,16a) were all superior in activity to resveratrol (1) as well as the remaining variations. Resveratrol (1), however, was comparable in activity to several of the other cis- stilbenes described here, including the 4'-hydroxy (14g) and the 3-hydroxy (14c) -derivatives.

For this reason the synthesis of the prodrug of the 4'-hydroxystilbene 14c and the 3- hydroxystilbene 14g was attempted. The synthesized prodrug 14n was essentially identical to 14c in its effects on the growth of cancer cells. From the known antimitotic activity of combretastatin A-4 (2a) and phenstatin (2c) and their potent interactions with tubulin, it seemed possible that the most cytotoxic compounds prepared in the cunent series would also inhibit this important cellular protein.

Several of the newly synthesized compounds were therefore examined for inhibitory effects on tubulin assembly, in a direct comparison with 2a and 2c. Compound 4a proved to be more inhibitory than 2a, while, in contrast, 16a was somewhat less potent than 2c. Resveratrol (1) was inactive as an inhibitor of tubulin assembly. Compound 14c, representing demethylation at position 3 of 4a, was 16-fold less active than 4a.

Combretastatin A-4 (2a) binds in the colchicine site of tubulin and is exceptionally potent as an inhibitor of the binding of radiolabeled colchicine to tubulin. (Lin, C. M., et al., Antimitotic natural products Combretastatin A-4 and Combretastatin A-2: Studies on the Mechanism of Their Inhibition of the Binding of Colchicine to Tubulin, Biochem. 1989, 28, 6984-6991.) The two new active compounds, 4a and 4c, were compared to 2a and phenstatin (2c) for their effects on colchicine binding to tubulin. Combretastatin A-4 (2a) displayed its usual potency, inhibiting colchicine binding by 98% when the two drugs were present in equimolar (5 μM) concentrations and by 91% when 2a was present at 2 μM (the tubulin

concentration in these experiments was 1.0 μM).

1398874.1/12504.012 26 Compound 4a was essentially equivalent to 2a as an inhibitor of colchicine binding. Phenstatin (2c), despite its greater inhibition of polymerization, was less potent than combretastatin A-4 (2a) as an inhibitor of colchicine binding, while 16a had the least activity in both assays. The reasons for these discrepancies are at present unknown but do not appear to derive from salt or temperature differences in the reaction condition.

There is also some indication that Resveratrol (1) contained in the roots of Polygonum cuspidatum has apparently been used in Chinese and Japanese traditional medicine as a treatment for gononhea. (Kubo, M., et al., Shoyayugaku Zasshi 1981, 35, 58.) In the present study, its activity against the etiologic agent of gononhea, Neisseria gonorrhoeae, was demonstrated in broth microdilution assays (Table IV). Stilbene production has been conelated with the resistance of grape leaves to fungal infection. (Langcake, R., et al., The Relationship of Resveratrol Production to Infection of Grapevine Leaves by Botrytis cenerea, Vitis 1979 18:244- 253.) Most of the stilbenes evaluated for antimicrobial action had antifungal activity (Table IV). The dimethyl derivative of cw-resveratrol (14c), with both antibacterial and antifungal activities, was the most active of the stilbenes and benzophenones tested (Table IV). Further biological evaluation of such resveratrol structural modifications should include chemopreventive potential targets such as COX-1 and COX-2.

1398874.1/12504.012 27 Table I. Cytotoxicity Data of Resveralrol (1), Combrclastalin A-4 (2a), Resvcraslalin (14c), the Oisodium Resverastatin Prodnig (14n) and Related Stilbenes.

Leukemia I'uncrcas-a Drcust CNS lung-NSC Colon I'loslalc

Compound P388 UXl'C-3 MCF-7 SF-268 NCI-11460 KM20L2 DU-145

EDjoμg/mL GIjoμg/mL

4.49 j .3 3. 9 4.1 3 . - TTT — I7T-

TT 24.4 1S . S 14.8 5.0 13.2 22.0 10.2

Ta 3.0x10^'* 0.39 - -1.0x10^"'' 6.0X10-' 0.34 8.0x10'

Ta 2.62xl0^'i 3.4xl0^"3 - 4.4X10^"1 2.8xl0^"J 5.4x10'

TIT 2.77 3 .5X10^'1 - 5.6x10-' 6.2x10^'' 1.5

ΈΓ 2.95 12.6 3 . 0 2.2 2.8 11.9 2.3

T?Γ 4.87 6.5 2.3 4.3 2.7 3.4 2.7

-149- 3.82 3.3 - 1.5 7.2x10" 2.6 2.5 lAh.

31.5 12.7 - 10.3 11.2 13.1 6.0

______

2.75 14.1 12.7 15.6 6.4 17.7 10.6

______

28.9 6.3 3 .4 3.1 6 .8 3 .8 2.7

______

4.45 5.4 9.0 38.5 10.7 24.7 35.6

1 1"

2.81 6.6 7.3X10^'1 1.9 3 .6 11.8 3.7

1398874.1/12504.012 28 Table II. Cytoloxicily Data ot Resveratrol (1), Phenstatin (2c) and Relaled Benzophenones and Benzhydrols.

Compound Leukemia Pαiicreas-a Breast CNS Lung-NSC Colon Pros

P388 BXPC-3 MCF-7 SH-268 NCI-H460 KM20L2 DU- l-D_₀μg/nιL GIjoμg/mL

4.49 3.3 3.9 4.1 3.6 13.1

2C 3.3xl0^'J - - 5.2x10"' 5.7xlO^'J 4.0X10"¹

Tϊn .

20.2 16.1 - 23.3 21.7 21.0 2

1.23 4.2X10^'1 - 8.6x10^"' 5.3x10" 4.2X10'¹

_L5c_

>10 1.3 2.0 6.5x10" 2.3 5.3

.fir

24.8 4.0 2.2 3.8 3.0 3.6

______

17.8 21.2 21.7 24.5 16.6 24.5 1

>100 20.5 30.6 22.3 25.0 24.5 1

■ fit

26.2 21.7 26.8 26.4 12.1 31.6 ]

_!£--_

13.4 15.0 14.2 13.9 10.3 16.2 ]

1398874.1/12504.012 29 Table III. Interactions of stilbenes .and benzophenones with tubulin. Compound Inhibition of tubulin Inhibition of colchicine

Polymerization binding

IC₅₀ (μM) ±SD % Inhibition

2μM inhibitor 5μM inhibitor

Resveratrol (1) >40

Combretastatin A-4 (2a) 2.0 ± 0.2 91 ± 3 98 ± 1

Phenstatin (2c) 1.1 ± 0.1 73 ± 1 85 ± 2

4a 1.8 ± 0.3 88 ± 1 95 ± 1

Resverastatin (14c) 29 ± 9

16a 2.6 ± 0.3 47 ± 5 69 ± 0.3

The tubulin polymerization assay was performed as previously described.²⁴ The tubulin concentration was 10 μM, with varying drug concentrations, as required, to obtain IC₅o values. Extent of assembly after 20 min at 30°C was the parameter measured. The colchicine binding assay was performed as earlier reported.²⁷ The tubulin concentration was 1.0 μM, the [3H] colchicine concentration was 5.0 μM, and the potential inhibitor concentrations were as indicated. Binding of colchicine was measured following a 10 min incubation at 37°C.

04.012 30 Table IV. Antimicrobial activities of resveratrol (1) and related stilbenes and benzophenones

Minimum inhibitory concentration (μg/ml)

Microorganism 1 4a 14c 14h 14k 14u 15c 16a 16c 16e I6g 16! 16k

Cryptocaccus neofonnaiis * * 16 16 * 64 * * * * * * 64

Candida albicans * * 64 ♦ * * * * * * * * *

Staphylocυccus aureus * * 32 * * * * * * * * * *

Streptococcus pneumoniae * * 16 * * * 16 * 64 * 64 64 32-64

Enterococcus faecalis * * 32-64 * * * * * * * *

Microcυccus luteus * * 8 16-64 • 32-64 + * * *

Escherichla coll * * * * * * * + * * * *

Enterococcus faecalis * « * * * * * * ♦ * ♦ * *

Enterobacter cloacae * * * * * + • * •*• * * *

Slenυtrυphυmonas * * * * * + * * * * * * * nialtophilia

Neisseria gonorrhoeae 16-32 * 8 8 16 8-32 2-4 32 32-64 * * 32 16-32

\ no inhibition at 64 μg/ml

4.1/12504.012 31

Claims

What is claimed is: 1. A resveratrol derivative having the formula:

selected from the group consisting of compounds wherein: a) Ri = R₂= R₃ = -OCH_3; and b) Ri = OH and R₂= R₃ = -OCH₃.

2. A resveratrol derivative having the formula:

wherein R₄ = R₅ = -OCH₃ and R^ = -OH.

3. A resveratrol derivative having the formula:

R*. wherein R₇ = R₈ = -OH and R₉ = -OCH₃.

1398874.1/12504.012 32

4. A resveratrol derivative having the formula:

selected from the group consisting of compounds wherein: a) Rio = Ri i = -OCH₃ and R_i2 = -O(PO)(OBn)₂; and b) R₁₀ = Rn = -OCH₃ and R_i2 = -O(PO)(ONa)₂.

5. A method for synthesizing the resveratrol derivative of claim 1 wherein Ri = R₂ = R₃ -OCH , comprising reacting phosphonium bromide and 3,5-dimethylbenzaldehyde.

6. A method for synthesizing the resveratrol derivative of claim 1 wherein Ri = -OH and R₂ = R₃ = -OCH₃, comprising the following steps:

5a 6

1398874.1/12504.012 33

9a

14b -*^■ 14d (£)

90%

14e,f 14g,

34

7. A method for synthesizing the resveratrol derivative of claim 2 comprising the following steps:

4a (Z),4b(£)

9a

1398874.1/12504.012 35

8. A method for synthesizing the resveratrol derivative of claim 3 comprising the following steps:

4a (2),4b(£)

9a

5b

1398874.1/12504.012 36

14b - 14d (E)

90%

141 (£)

37

9. A method for synthesizing the resveratrol derivative of claim 4 wherein Rio = Ri ι = -OCH₃ and Rι₂ = -O(PO)(OBn)₂, comprising the following steps:

4a (Z),4b(£)

9a

1398874.1/12504.012 38

14b * 14d (£)

90%

14e,f 14g,h

39

141 (£)

10. A method for synthesizing the resveratrol derivative of claim 4 wherein Rio = Ri ₁ -OCH3 and -O(PO)(ONa)₂, comprising the following steps:

Λa (Z),4b(-=)

SPD

9a

1398874.1/12504.012 40

^14b „_»B ' 4 (E) 90%

141 (£)

1398874.1/12504.012 41

11. A pharmaceutical composition comprising the compound of claim 1, or a pharmaceutically acceptable salt thereof.

12. A pharmaceutical composition comprising the compound of claim 2, or a pharmaceutically acceptable salt thereof.

13. A pharmaceutical composition comprising the compound of claim 3, or a pharmaceutically acceptable salt thereof.

14. A pharmaceutical composition comprising the compound of claim 4, or a pharmaceutically acceptable salt thereof.

15. A method for treating humans and mammals afflicted with cancer, comprising administering a physiologically effective amount of the compound of claim 1, or a pharmaceutically acceptable salt thereof.

1398874.1/12504.012 42

16. A method for treating humans and mammals afflicted with cancer, comprising administering a physiologically effective amount of the compound of claim 2, or a pharmaceutically acceptable salt thereof.

17. A method for treating humans and mammals afflicted with cancer, comprising administering a physiologically effective amount of the compound of claim 3, or a pharmaceutically acceptable salt thereof.

18. A method for treating humans and mammals afflicted with cancer, comprising administering a physiologically effective amount of the compound of claim 4, or a pharmaceutically acceptable salt thereof.

1398874.1/12504.012 43