Substituted trans-stilbenes, including analogues of the natural product resveratrol, inhibit the human tumor necrosis factor alpha-induced activation of transcription factor nuclear factor kappaB

Article abstract of DOI:10.1021/jm060630x

The transcription factor nuclear factor kappaB (NF-κB), which regulates expression of numerous antiinflammatory genes as well as genes that promote development of the prosurvival, antiapoptotic state is up-regulated in many cancer cells. The natural product resveratrol, a polyphenolic trans-stilbene, has numerous biological activities and is a known inhibitor of activation of NF-κB, which may account for some of its biological activities. Resveratrol exhibits activity against a wide variety of cancer cells and has demonstrated activity as a cancer chemopreventive against all stages, i.e., initiation, promotion, and progression. The biological activities of resveratrol are often ascribed to its antioxidant activity. Both antioxidant activity and biological activities of analogues of resveratrol depend upon the number and location of the hydroxy groups. In the present study, phenolic analogues of resveratrol and a series of substituted trans-stilbenes without hydroxy groups were compared with resveratrol for their abilities to inhibit the human tumor necrosis factor alpha-induced (TNF-α) activation of NF-κB, using the Panomics NF-κB stable reporter cell line 293/NF-κB-luc. A series of 75 compounds was screened to identify substituted trans-stilbenes that were more active than resveratrol. Dose-response studies of the most active compounds were carried out to obtain IC₅₀ values. Numerous compounds were identified that were more active than resveratrol, including compounds that were devoid of hydroxy groups and were 100-fold more potent than resveratrol. The substituted trans-stilbenes that were potent inhibitors of the activation of NFκB generally did not exhibit antioxidant activity. The results from screening were confirmed using BV-2 microglial cells where resveratrol and analogues were shown to inhibit LPS-induced COX-2 expression.

Full text of DOI:10.1021/jm060630x

Supporting Information
Substituted trans-Stilbenes, Including Analogs of the Natural Product Resveratrol,
Inhibit the Human Tumor Necrosis Factor Alpha-induced Activation of
Transcription Factor Nuclear Factor-kappa B
Justin J. Heynekamp,† Waylon M. Weber,† Lucy A. Hunsaker,‡ Amanda M. Gonzales, ‡
Robert A. Orlando, ‡ Lorraine M. Deck,*,† and David L. Vander Jagt*,‡
Department of Chemistry, University of New Mexico, Albuquerque, NM 87131;
Department of Biochemistry and Molecular Biology, University of New Mexico School
of Medicine, Albuquerque, NM 87131
* Corresponding authors: LMD, phone 505-277-5438; Email: ldeck@unm.edu; DLVJ,
phone 505-272-5788; Email: dlvanderjagt@salud.unm.edu
† Department of Chemistry, University of New Mexico
‡ Department of Biochemistry and Molecular Biology, University of New Mexico School
of Medicine
CONTENTS: Supporting information includes general synthetic procedures, list of
known compounds that were included in this study along with references.
S1

Unless otherwise noted all reagents were obtained from commercial sources and
used without further purification. All compounds that were isolated were greater than
13
90% pure by 1H and/or C NMR. Column chromatographic separations were performed
using EM Science type 60 silica gel (230-400 mesh). Melting points were determined on
a Thomas Hoover capillary melting point apparatus and are uncorrected. NMR spectra
were recorded on a Bruker AC250 (250 MHz) NMR spectrometer in CDCl3 unless
otherwise noted. Chemical shifts are reported in ppm (δ) relative to CHCl3 at 7.24 ppm
for 1H NMR and 77.0 for 13C NMR. High resolution mass spectra were performed at the
Mass Spectrometry Facility, University of New Mexico.
General method for the preparation of substituted benzyl phosphonic acid diethyl esters.
Substituted benzyl bromide was heated with excess triethylphosphite at 140ºC until the
evolution of bromoethane had ceased and complete dissolution occurred. The remaining
triethylphosphite was then removed by concentration of the solution in vacuo to afford the
product.
General method for the preparation of MOM protected aldehydes.
To a suspension of hexane rinsed sodium hydride (1.5 equivalents) in dimethyl formamide is
added a solution of the appropriate aldehyde (1 equivalent) in dimethyl formamide. After
stirring 2 hours at room temperature dichloromethylmethyl ether is added and the solution is
stirred an additional 3 hours at room temperature. The solution is quenched by pouring over
ice water and extracted with ether. The ether extracts are washed with 1M sodium hydroxide,
saturated sodium chloride and dried with magnesium sulfate, filtered and evaporated to give a
crude oil that is distilled bulb to bulb to afford the product as a solid.
S2

(E)-3,4-Dimethoxystilbene (4a). mp 108-109 °C [lit.1 112-113 °C].
(E)-4-Methoxystilbene (4b). mp 134-135 °C [lit.2 135-136 °C].
(E)-4-Chlorostilbene (4c). mp 129-130 °C [lit.3 129-131 °C].
(E)-4-Methylstilbene (4d). mp 114-116 °C [lit.3 118-120 °C].
(E)-4-Cyanostilbene (4e). mp 116-117 °C [lit.3 115-118 °C].
(E)-3,5-Dimethoxystilbene (4f). mp 53-55 °C [lit.4 54-55 °C].
(E)-3-Chlorostilbene (4h). mp 74-76 °C [lit.5 71-72.5 °C].
(E)-3-Methylstilbene (4i). mp 50-51 °C [lit.6 48-49 °C].
(E)-2-Chlorostilbene (4j). mp 64-66 °C [lit.7 37-38 °C].
(E)-4-Ethoxystilbene (4k). mp 124-126 °C [lit.8 77-78 °C].
(E)-4-Hydroxystilbene (4l). mp 183-185 °C [lit.9 188 °C].
(E)-4-Fluorostilbene (4m). mp 123-124 °C [lit.10 124 °C].
(E)-3-Fluorostilbene (4n). mp 74-76 °C [lit.11 70-72 °C].
(E)-2,3-Dimethoxystilbene (4o). mp 37-39 °C [lit.12 38-39 °C].
S3

(E)-2-Fluorostilbene (4p). mp 103-105 °C [lit.13 102-103 °C].
(E)-4-Hydroxy-3-methoxystilbene (4q). mp 133-134 °C [lit.9 138 °C].
(E)-3-Methoxystilbene (4r). mp 34-35 °C [lit.2 34-35 °C].
(E)-3-Hydroxystilbene (4t). mp 119-121 °C [lit.14 119-120 °C].
(E)-2,4-Dimethoxystilbene (4u). mp 64-65 °C [lit.15 64.5-65 °C].
(E)-2-Methylstilbene (4v). mp 35-36 °C [lit.2 28-29 °C].
(E)-3-Trifluoromethylstilbene (4w). mp 67-68 °C [lit.16 66-67 °C].
(E)-4-Trifluoromethylstilbene (4x). mp 133-134 °C [lit.2 134-135 °C].
(E)-2,5-Dimethoxystilbene (4y). oil; 1H NMR: δ 3.81 (s, 3H), 3.83 (s, 3H), 6.80 (m, 2H),
7.09 (d, 1H, J = 16.48 Hz), 7.16 (d, 1H, J = 2.38 Hz), 7.25 ( m, 1H), 7.34 (t, 2H, J = 7.35
Hz), 7.47 (d, 1H, J= 16.49 Hz), 7.53 (d, 2H, J = 7.15 Hz).
(E)-2,4,6-Trimethylstilbene (4aa). mp 56-57 °C [lit.17 49-50 °C].
(E)-2-Methoxystilbene (4bb). mp 58-59 °C [lit.2 56-57 °C].
(E)-4-N,N-Dimethylaminostilbene (4cc). mp 144-146 °C [lit.18 150 °C].
S4

(E)-3,4-Dihydroxystilbene (4ee). mp 167-168 °C [lit.19 168-169 °C].
(E)-3,4,5-Trimethoxystilbene (4ff). mp 107-108 °C [lit.20 105-106 °C].
(E)-2,3,4-Trimethoxystilbene (4gg). mp 80-83 °C [lit.20 79-82 °C].
(E)-3,4,4’-Trimethoxystilbene (6a). mp 136-138 °C [lit.4 136-138 °C].
(E)-4,4’-Dimethoxystilbene (6b). mp 212-213 °C [lit.2 214-216 °C].
(E)-4-Chloro-4’-methoxystilbene (6c). mp 181-183 °C [lit.4 181-184 °C].
(E)-4’-Methoxy-4-methylstilbene (6d). mp 160-162 °C [lit.21 166-167 °C].
(E)-4-Cyano-4’-methoxystilbene (6e). mp 141-143 °C [lit.22 141-142 °C].
(E)-3,4’,5-Trimethoxystilbene (6f). mp 53-55 °C [lit.4 53-56 °C].
(E)-3-Chloro-4’-methoxystilbene (6h). mp 93-94 °C [lit.21 96 °C].
(E)-4’-Methoxy-3-methylstilbene (6i). mp 110-111 °C [lit.21 98 °C].
(E)-2-Chloro-4’-methoxystilbene (6j). mp 52-53 °C [lit.23 59-60 °C].
(E)-4-Ethoxy-4’-methoxystilbene (6k). mp 194-195 °C [lit.24 165-167 °C].
(E)-3,4,4’,5-Tetramethoxystilbene (6l). mp 157-159 °C [lit.20 152-155 °C].
S5

(E)-4-Fluoro-4’-methoxystilbene (6m). mp 148-150 °C [lit.25 147-149 °C].
(E)-3-Fluoro-4’-methoxystilbene (6n). mp 108-110 °C [lit.11 108-110 °C].
(E)-2,3,4’-Trimethoxystilbene (6o). mp 70-72 °C [lit.26 73-74 °C].
(E)-2-Fluoro-4’-methoxystilbene (6p). mp 100-101 °C [lit.13 102-103 °C].
(E)-2,4,4’,5-Tetramethoxystilbene (6q). mp 106-107 °C [lit.27 110 °C].
(E)-3,4’-Dimethoxystilbene (6r). mp 107-108 °C [lit.26 107-108 °C].
(E)-4-Bromo-4’-methoxystilbene (6t). mp 200-201 °C [lit.28 177-179 °C].
(E)-2,4,4’-Trimethoxystilbene (6u). mp 94-95 °C [lit.29 89 °C].
(E)-3,4-Dihydroxy-4’-methoxystilbene (6x). mp d 186 °C [lit.30]; 1H NMR: (DMSO-d6)
δ 3.75 (s, 3H), 6.70 (d, 1H, J = 8.94 Hz), 6.87 (m, 6H), 7.45 (d, 2H, J = 8.34 Hz), 8.88 (s,
1H), 9.00 (s, 1H).
1
(E)-2,4’,5-Trimethoxystilbene (6y). mp 67-68 °C [lit.31 oil]; H NMR: δ 3.80 (s, 3H),
3.81 (s, 3H), 3.82 (s, 3H), 6.80 (m, 2H), 6.88 (d, 2H, J = 8.74 Hz), 7.04 (d, 1H, J = 16.48
Hz), 7.13 (d, 1H, J = 2.78 Hz), 7.32 (d, 1H, J = 16.48 Hz), 7.47 (d, 2H, J = 8.74 Hz).
(E)-2,4’-Dimethoxystilbene (6bb). mp 89-90 °C [lit.32 85-86 °C].
S6

(E)-4’-Methoxy-4-N,N-dimethylaminostilbene (6dd). mp 182-183 °C [lit.33 185-186
°C].
(E)-2,6-Dichloro-4’-methoxystilbene (6ee). mp 56-60 °C; 1H NMR: δ 3.83 (s, 3H), 6.92
(d, 2H, J = 8.74 Hz), 7.05 (m, 3H), 7.33 (d, 2H, J = 7.94 Hz), 7.49 (d, 2H, J = 8.54 Hz).
(E)-3-Stilbazole (8a). mp 81-82 °C [lit.34 83-85 °C].
(E)-4-Stilbazole (8b). mp 124-126 °C [lit.35 128 °C].
(E)-2-Stilbazole (8c). mp 89-91 °C [lit.36 93 °C].
(E)-2-Styrylthiophene (8d). mp 111-112 °C [lit.36 112-113 °C].
(E)-2-Styrylnaphthalene (8e). mp 147-148 °C [lit.37 147-148 °C].
(E)-1-Styrylnaphthalene (8f). mp 71-72 °C [lit.38 71-72 °C].
(E)-2-(4-Methoxystyryl)naphthalene (8g). mp 172-173 °C [lit.39 142 °C].
(E)-1-(4-Methoxystyryl)naphthalene (8h). mp 93-94 °C [lit.40 92-93 °C].
(E)-2-(4-Methoxystyryl)thiophene (8i). mp 133-134 °C [lit.41 134-135 °C].
(E)-4’-Methoxy-3-stilbazole (8j). mp 98-100 °C [lit.42 99-100 °C].
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