Simple and efficient synthesis of benzofuran derivatives from tyrosol

Article abstract of DOI:10.1080/00397911.2015.1133828

A convenient strategy for the preparation of compounds bearing the benzofuran skeleton starting from tyrosol, a phenol largely present in olive oil production waste, with no biological importance, is reported. A bromination/methoxylation sequence, already described for the synthetic transformation of naturally occurring compounds, was exploited. Depending on the solvent used for the methoxylation reaction together with the presence of a 4-phenol moiety respect to the side chain, benzodihydrofurans or a benzofurans derivative can be obtained.

Full text of DOI:10.1080/00397911.2015.1133828

Synthetic Communications
An International Journal for Rapid Communication of Synthetic Organic
Chemistry
ISSN: 0039-7911 (Print) 1532-2432 (Online) Journal homepage: http://www.tandfonline.com/loi/lsyc20
Simple and Efficient Synthesis of Benzofuran
Derivatives from Tyrosol
Paolo Bovicelli, Fabrizio Bottaro, Carla Sappino, Michela Tomei, Valentina
Nardi, Ilaria Proietti Silvestri, Beatrice Macchi, Caterina Frezza & Giuliana
Righi
To cite this article: Paolo Bovicelli, Fabrizio Bottaro, Carla Sappino, Michela Tomei, Valentina
Nardi, Ilaria Proietti Silvestri, Beatrice Macchi, Caterina Frezza & Giuliana Righi (2015): Simple
and Efficient Synthesis of Benzofuran Derivatives from Tyrosol, Synthetic Communications,
DOI: 10.1080/00397911.2015.1133828
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Date: 02 February 2016, At: 01:34

SIMPLE AND EFFICIENT SYNTHESIS OF BENZOFURAN DERIVATIVES
FROM TYROSOL
Paolo Bovicelli1,, Fabrizio Bottaro1, Carla Sappino1, Michela Tomei1, Valentina Nardi1,
Ilaria Proietti Silvestri1, Beatrice Macchi2, Caterina Frezza2, Giuliana Righi1
1C.N.R. Institute of Biology and Molecular Patologies (IBPM), Department of
Chemistry, Sapienza University of Rome, Rome, Italy
2Department of System Medicine, “Tor Vergata” University, Rome, Italy
Address correspondence to Paolo Bovicelli . E-mail: paolo.bovicelli@cnr.it
Abstract
A convenient strategy for the preparation of compounds bearing the benzofuran skeleton
starting from tyrosol, a phenol largely present in olive oil production waste, with no
biological importance, is reported. A bromination/methoxylation sequence, already
described for the synthetic transformation of natural occurring compounds, was
exploited. Depending on the solvent used for the methoxylation reaction together with the
presence of a 4-phenol moiety respect to the side chain, benzodihydrofurans or a
benzofurans derivative can be obtained.
KEYWORDS: Benzofurans; polyphenols; antioxidants; heterocyclization
INTRODUCTION

In the last years we have largely investigated a method to oxyfunctionalize the aromatic
ring of natural occurring compounds in order to enhance their biological activities.[1] The
method involves a methoxylation of an aromatic bromide, a reaction optimized by us and
already reported.[2] The bromination of activated aromatic rings can be performed by a
number of methods,[3] but we found the system NaBr/Oxone to be the most efficient and
clean. In this way the bromination can be modulated to easily obtain the desired products.
The most interesting example is the bromination of tyrosol (4-hydroxyphenethyl alcohol)
which can be selectively transformed to the monobromo or dibromo derivatives, allowing
to access to aromatic compounds with different degree of oxygenation.
The main example of the potentiality of this synthetic pathway for the preparation of
compounds bearing the benzofuran skeleton is the easily and convenient transformation
of tyrosol into hydroxytyrosol.[4]
Other methods are reported in literature for etherification of aryl bromides, by catalysis of
palladium using complex and costly ligands,[5] or copper under conditions which extend
our method,[6] but our goal is to exploit this strategy to produce compounds with high
value starting from waste of industry. Tyrosol is a polluting chemical, largely present in
the waste of the olear industry, with no biological importance, meanwhile hydroxytyrosol
is one of the strongest antioxidants in nature, with many potential uses in medicine, as
integrator, in protective and anti-age creams, and more.[7] By this methodology we were
able to conveniently produce rare natural compounds.[8] The advantage is to make those
compounds available for biological investigations at low cost.

RESULTS AND SISCUSSION
In this work we describe a facile and straightforward synthesis of benzofuran derivatives
starting from tyrosol using the bromination/methoxylation approach. The synthetic
pathway for obtaining benzofuran and benzodihydrofuran derivatives is summarized in
Scheme 1. Such as for the transformation of tyrosol into hydroxytyrosol, the aim of this
work is to valorize an industrial waste.
The first step of the synthesis is a monobromination of tyrosol, which was performed as
previously reported[4] paying attention to the reaction conditions to avoid the further
halogenation. In an improved procedure with respect to that previously reported by us,[4]
the methoxylation was performed in DMF at 100°C stirring overnight, but we observed
that the reaction occurs also in eco-sustainable dimethylcarbonate (DMC) at lower
temperature, namely 90°C.[1]
Our purpose is always to use conditions with lower environmental impact, but in the first
methoxylation step the use of DMC as solvent was not possible due to the competing
carboxymethylation of the side chain and this was not compatible with further steps.
The second bromination was performed on the homovanillic alcohol protected as
diacetate (4) to avoid its decomposition in the oxidative media.
Finally, the ring closure was obtained under the same conditions as the methoxylation.
The probable mechanism of the reaction involves an oxidative addition of the copper to

the aromatic ring. The primary alcohol of the side chain can act as competitive ligand for
the metal leading to an intramolecular insertion after the reductive elimination. In this
case the process is so favorable that it occurs at lower temperature.
The reaction was carried out in DMC and good yield of the benzodihydrofuran derivative
was obtained. In more drastic conditions (DMF, 120°C, 8 hrs) the reaction shifted
towards the benzofuran derivative 9. This occurs only when a para free phenol moiety
with respect to the side chain is present. Although the mechanism at the basis of the
process is still under investigation a way to produce 6 or 9 selectively is now available.
Compounds 7 and 8 were obtained by methylation and demethylation of 6 respectively.
10 was produced from 9 by methylation meanwhile the demethylation reaction led to a
complex mixture of products, due to the liability of catechol compound.
By this pathway, a number of variously substituted benzodihydrofurans and benzofurans
can be synthesised staring from tyrosol. An example is reported in Scheme 2 for
compounds 15 and 16 with a fully oxygenated aromatic ring. In the methoxylation of 14
the two bromines were substituted by the ethanolic chain and the methoxy group
respectively. The ring closure was complete before the methoxylation step since the
monobrominated dihydrobenzofuran can be isolated.
BIOLOGICAL ASSAYS

The biological activity of 6, 7, 8, 9, 10, 15Ac, 16Ac, 18 was assessed by measuring their
ability to induce cytotoxicity by inhibiting cell metabolism and cell death by apoptosis in
a monocytoid cell line U937. Toxicity of the drugs was evaluated by inhibition of cell
metabolic activity, measured by reduction of MTS (3-(4,5-dimethylthiazol-2-yl)-5- (3-
carboxymethoxyphenyl)-2- (4-sulfophenyl)-2H tetrazolium) to formazan, using a
commercial colorimetric as previously described.[9] The results were expressed as the
drug concentration required to inhibit 50 % of the metabolic activity (CC50).Apoptosis
was expressed as percentage of hypodiploid nuclei following staining with Propidium
iodide (PI), and evaluation through cytofluorimetric analysis.[1][0] The results in Table 1,
show that the compounds used at the concentration of 250 µM, variably induced in vitro
cell death by apoptosis in U937 cells. In particular the compound 8, was the most
efficacious in inducing 71% of hypodiploid nuclei, while 9 and 15Ac induced 10% and
19% hypodiploid nuclei respectively. However 8, 9 and 15Ac, were between 25 and 200
times less active than the chemotherapeutic drug etoposide, SN38 used as reference
control. On the other side all the tested compounds did not highly inhibit the metabolic
activity in U937 cells, showing IC50 between 1000 and 500 µM, except 8 that exhibited
IC50 of 253 µM. Taken together these data indicated that these compounds were less
toxic in vitro in comparison with SN38 (IC50 14 µM), nevertheless their pro apoptotic
activity was lower than the reference compound. Therefore this class of molecules could
be lead compounds to be undergoing further modification aimed to increase their
specific activity.
CONCLUSIONS

In this work a synthetic pathway to obtain bicyclic oxygenated compounds from tyrosol
is reported. In DMC and controlling reaction times benzodihydrofurans can be selectively
obtained. Their dehydrogenated derivatives can be produced using DMF as the solvent
and prolonging the reaction time.
All polyoxygenated compounds easily prepared by the reported methodology were
investigated for their potential biological properties.
EXPERIMENTAL
General Procedures.
Bromination reaction. In a 50 mL round bottom flask equipped with magnetic stirring 1
mmol of substrate and 1 mmol of NaBr (102 mg) were dissolved in 7.5 mL of acetone. A
solution of 900 mg of oxone in 7.5 mL of water was slowly added under stirring until
completion of reaction (monitoring by TLC). 3 mL of saturated solution of sodium
thiosulfate in water were then added. Acetone was evaporated under vacuum and the
aqueous layer was extracted with ethyl acetate (3 x 20 mL). The organic layer was
washed with brine (3 x 15 mL) and dried with anhydrous sodium sulfate and the solvent
evaporated under vacuum. Normally the product was utilized for the further reactions
without any other purification.
The same procedure was used to obtain the dibrominated analog, increasing the
equivalents of NaBr and oxone.

Methoxylation reaction. In a 25 mL round bottom flask an excess of complex solution
was prepared dissolving in DMF (or DMC) 168 mg of CuBr and 10 mL of 25% w
solution of CH3ONa in methanol. After few minutes under stirring the mixture became
blue. This solution was added in small portions to a solution of substrate (1 mmol) in
DMF or DMC (2 mL) heated at the appropriate temperature (90°C for DMC and 100°C
for DMF). The reaction was monitored by TLC, at the end the mixture was cooled in an
ice bath and acidified with a 2M HCl solution. Methanol was evaporated under reduced
pressure and the residue extracted with Et2OAc (3 x 20 mL), washed with brine (3 x 15
mL), dried with anhydrous Na2SO4 and the solvent evaporated under vacuum. The crude
was purified by flash chromatography (hexane/Et2OAc 4:1).
Methylation reaction. 1 mmol of substrate was dissolved in 7 mL of acetone, 3 mmol of
K2CO3 and 2 mmol of dimethyl sulfate were added for every phenol moiety. The mixture
was stirred overnight at r.t. The reaction was quenched with a 10% solution of NH4OH in
water (5 mL) then acetone was removed under vacuum. The residue was dissolved with
Et2OAc and washed with a 2M solution of HCl in water until acidity. The product was
obtained in substantially quantitative yield.
Demethylation reaction. BBr3 (4.5 mL) was added to 1 mmol of substrate dissolved in
dichloromethane (7 mL) at -20°C. After the reaction was complete, monitoring it by
TLC, the mixture was quenched with water. The crude was extracted with diethyl ether (3
x 10 mL), the organic layers was washed with brine (3 x 5 mL) and dried with anhydrous

Na2SO4. After evaporation of the solvent under vacuum the product was obtained pure
enough to be used for further reactions.
In example compounds 8 and 10 were synthetized through a sequence of reaction as
above.
2,3-Dihydro-benzofuran-5,6-diol (8). Light brown solid, 74%. M.p. 107.2-109°C. IR
(CHCl3, cm-[1]): 3210, 1615, 1275, 840. [1]H -NMR D6-DMSO ppm): 6.92 (1H, s, ArH),
6.72 (1H, s, ArH), 4.86 (3H, s, OCH3), 3.67 (2H, t, J=7.3 Hz, CH2O), 2.79 (2H, t, J=7.3
Hz, CH2Ar). [1][3]C -NMR D--DMSO ppm,): 144.6, 144.5, 128.6, 118.7, 117.3, 112.1,
61.4, 38.2. Analysis, calc. for C8H8O3: C 63.15, H 5.30; found C 63.12, H 5.27.
5,6-Dimethoxy-benzofuran (10). White solid, 35%. M.p. 54.5-56.5°C. [1]H -NMR
ppm): 7.50 (1H, d, J=1.9 Hz, C=CH-O), 7.04 (1H, s, ArH), 7.00 (1H, s, ArH), 6.64
(1H, d, J=1.9 Hz, C=CH), 3.89 (3H, s, OCH3). [1][3]C -NMR ppm): 149.5, 147.8, 146.4,
143.8, 119.0, 106.4, 102.2, 95.2, 56.2, 56.0. Analysis, calc. for C10H10O3: C 67.41, H
5.66; found: C 67.42, H 5.64.
ACKNOWLEDGMENTS
We thank for financial support MIUR (Ministry of University and Research – Rome)
(PRIN 2010-2011: Design and stereoselective synthesis of compounds active towards
proteic targets involved in viral and tumoral pathologies) and Italian Ministry for
Education, University and Research in the framework of the Flagship Project NanoMAX.

SUPPLEMENTAL MATERIAL
Supplemental data for this article can be accessed on the publisher’s website.
REFERENCES
1. Barontini, M.; Proietti Silvestri, I.; Nardi, V.; Crisante, F.; Pepe, G.; Pari, L.; Gallucci,
F.; Bovicelli, P.; Righi, G. Med. Chem. Res. 2013, 22(2), 674-680.
2. Bovicelli, P.; Antonioletti, R.; Barontini, M.; Borioni, G.; Bernini, R.; Mincione, E.
Tetrahedron Lett. 2005, 46(8), 1255-1257.
3. Sternbach, L.H. Prog. Drug Res. 1978, 44, 1340–1344. Goldberg, Y.; Alper, H. J. Mol.
Cat. 1994, 88, 377. Roy, S.C.; Guin, C.; Rana, K.K.; Maiti, G. Tetrahedron Lett. 2001,
42, 6941–6942.
4. Bovicelli, P.; Antonioletti, R.; Mancini, S.; Causio, S.; Borioni, G.; Ammendola, S.;
Barontini, M. Synth. Commun. 2007, 37(23), 4245-4252.
5. Kubawe, S.; Torraca, K. E.; Buchwald, S. L. J. Am. Chem. Soc. 2000, 122, 12907-
12908; 2001, 123, 12202-12206.
6. Huang, J.; Chen, Y.; Chan, J.; Ronk, M. L.; Larsen, r. D.; Faul, M. M. Synlett 2011, 10,
1419-1422.
7. De Leonardis, A.; Aretini, A.; Alfano, G.; Macciola, V.; Ranalli, G. Eur. Food Res.
Technol. 2008, 226(4), 653–659. Achmon, Y. ; Fishman, A. App. Microb. Biotec. 2015,
99(3), 1119-1130.
8. Bovicelli, P. J. Pharm. Pharmac. 2007, 59(12), 1703-1710.

9. Balestrieri, E.; Pizzimenti, F.; Ferlazzo, A.; Giofre, S.V.; Iannazzo, D.; Piperno, A.;
Romeo, R.; Chiacchio, M.A.; Mastino, M.; Macchi, B. Bioorg. & Med. Chem. 2011, 19,
2084-2089.
10. Matteucci, C.; Minutolo, A.; Balestrieri, E.; Marino-Merlo, F.; Bramanti, P.; Garaci,
E.; Macchi B.; Mastino, A. Cell Death in Dis. 2010, 1, 1-8.

Table 1. a) Apoptosis was evaluated in U937 cells as a percentage of hypodiploid nuclei
by flow cytometry analysis after 24 h of incubation. b) MACC50 is the metabolic activity
cytotoxic inhibitory concentration 50%, evaluated in the U937 cell line by MTS assay
Compound
Conc. (µM)
250
% hypodiploid nuclei(a)
MACC50 (µM)(b)
6
5
818
7
250
4
> 1000
253
8
250
71
10
7
9
250
744
10
250
737
15Ac
16Ac
18
250
19
4
557
250
> 1000
803
250
9
SN38
10
80
14

Scheme 1. Synthesis of dioxygenated benzofuran derivatives

Scheme 2. Synthesis of tetraoxygenated benzofuran derivatives