Styrylquinazoline derivatives as HIV-1 integrase inhibitors

Article abstract of DOI:10.1002/1521-4184(200208)335:6<277::AID-ARDP277>3.0.CO;2-A

Styrylquinazoline derivatives were prepared by Perkin condensation and evaluated for inhibitory activity against HIV-1 integrase. Among them, compound 5 c containing a free catechol ring was the most potent (IC₅₀ = 20.8 ± 1.9 μM) and showed 6-fold more potency than the corresponding styrylquinoline compound (IC₅₀ = 130.7 ± 8.6 μM).

Full text of DOI:10.1002/1521-4184(200208)335:6<277::AID-ARDP277>3.0.CO;2-A

Arch. Pharm. Pharm. Med. Chem. 2002, 6, 277–282
Styrylquinazoline Derivatives277
Jae Yeol Leea,
Styrylquinazoline Derivatives as HIV-1 Integrase
Inhibitors
Jang Hyun Parkb,
Sook Ja Leeb,
Hokoon Parka,
Yong Sup Leea
Styrylquinazoline derivativeswere prepared by Perkin condensation and evaluated
for inhibitory activity against HIV-1 integrase. Among them, compound 5 c contain-
a
Medicinal Chemistry
Research Center,
ing a free catechol ring wasthe most potent (IC
= 20.8 1.9 µM) and showed
50
Korea Institute of Science
& Technology, Cheongryang,
Seoul, Korea
6-fold more potency than the corresponding styrylquinoline compound (IC50
130.7 8.6 µM).
=
b
Department of Chemistry,
Hankuk University of
Foreign Studies, Yong-in,
Korea
Keywords: HIV; Integrase inhibitor; Styrylquinazoline; Perkin condensation; AIDS
Received: November 5, 2001 [FP647]
Introduction
The alarming spread of the acquired immune deficiency
syndrome (AIDS) epidemic has stimulated the discovery
of therapeutic agentsto inhibit the replication of the
causative virus, human immunodeficiency virus (HIV)
[1]. The advanced understanding of the viral cell cycle
has made it possible to define the targets to interrupt the
life cycle of the virus.Among them, one such target is the
viral integrase which is responsible for the integration of
proviral DNA into the host cell DNA. It catalyzes two dis-
tinct reactions: terminal cleavage at each 3Ј end of the
proviral DNA removing a pair of bases and strand trans-
fer which results in the joining of each 3Ј end to 5Ј-phos-
phatesin the target DNA [2]. Asthese reactionsare es-
sential for the life cycle of the virus, integrase represents
an attractive target for treatment of HIV infections.While
a large number of compoundsthat inhibit integrase have
already been identified [3], only a handful displayed anti-
viral activity in cell culture. To date, only G-rich oligonu-
cleotide (Zentivir), which was shown to inhibit integrase
activity both in vitro and in vivo, hasbeen undergoing
phase I/II clinical studies in AIDS patients [4]. However,
the clinical use of such an oligonucleotide has a limit be-
cause there is a problem of the capacity to produce
amountsof the compound compatible with therapeutic
exigencies[5].
Figure 1
Based upon this result and the various biological activi-
ties of the quinazoline ring system [7], we set out to syn-
thesize styrylquinazoline derivatives (II) as isosters of
styrylquinolines (I), with a view to inves tigating the effect
of the quinazoline subunit on the inhibitory activity
against HIV integrase and develop structurally simple in-
hibitorsof HIV integrase (Figure 1). In thispaper, we will
describe the synthesis of new styrylquinazoline deriva-
tives( II) and the inhibitory activity against HIV-1 inte-
gras e in 3Ј-processing reaction.
Recently, it wasfound that the styrylquinoline derivatives
(I) inhibited HIV-1 integrase in vitro and blocked HIV-1
replication in CEM cellsat nontoxic concentrationsas
shown in Figure 1 [6].
Correspondence: Yong Sup Lee, Medicinal Chemistry
Research Center, Korea Institute of Science & Technology,
P.O. Box 131 Cheongryang, Seoul 130-650, Korea.Phone:+82
2 958 5004, Fax: +82 2 958 5189, e-mail: yslee@kist.re.kr
© WILEY-VCH Verlag GmbH, 69451 Weinheim, 2002
0365-6233/06/0277 $ 17.50+.50/0

278 Lee et al.
Arch. Pharm. Pharm. Med. Chem. 2002, 6, 277–282
[8]. The Perkin condensation between 2-methylquinaz-
oline (2 a or 2 b) and variousaromatic aldehydes( 3) gave
styrylquinazolines (4 a–f) with pure E geometry under
reflux condition of acetic anhydride in 27–47 % yield, and
all hydroxy groupsof compounds 4 a–f were concomi-
tantly acetylated during thisreaction [9]. In the mean-
while, it iswell known to date that the most potent HIV in-
tegrase inhibitorssuch asL-chicoric acid and 4,5-di-
caffeoylquinic acid (4,5-DCQA) generally contain a free
Results and discussion
Chemistry
Two 2-methylquinazolines( 2 a: R1 = H, 2 b: R1 = OCH3)
asthe starting materialswere easily prepared from an-
thranilic acids( 1 a: R1 = H, 1 b: R1 = OCH3), respectively,
by known proceduresin order to investigate the effect of
the C-8 substituent on the quinazoline ring as well as the
quinazoline scaffold on the inhibitory activity (Scheme 1)
Table 1. Structuresand yieldsof styrylquinazoline derivatives( 4 a–f, 5 a–f, 6 a, and 6 b).
Scheme 1

Arch. Pharm. Pharm. Med. Chem. 2002, 6, 277–282
catechol-type aromatic moiety [10]. Therefore, the hy-
Styrylquinazoline Derivatives279
hydroxy group (5 a, 5 d, and 6 a) were als o inactive irre-
drolysisof acetoxy groupsof styrylquinazolines(
4 a–f)
spective of C-8 substituents of quinazoline ring.
wasaccomplished with pyridine/water or 0.5 N NaOH
(aq) to afford free hydroxystyrylquinazolines (5 a–f) in
21–87 % yield.With a view to testing the effect of another
“free” hydroxy group of the quinazoline ring on the inhibi-
tory activity against HIV-1 integrase, we also accom-
plished BBr3-induced demethylation of C-8 methoxy
group of compounds 5 d and 5 f to provide 8-hydroxy-
quinazolines( 6 a and 6 b) in 21 and 24 % yield, respec-
tively, asshown in Scheme 1 [11].The yieldsof all reac-
tionswere summarized in Table 1.
The active compounds( 5 c, 5 f, and 6 b), in which a free
catechol ring is present, showed good inhibitory activi-
tieswith IC
valuesof 20.08 1.9, 31.6 6.1, and
50
57.1 9.3 µM, respectively.This observation means that
the free catechol moiety of styrylquinazoline compounds
isrequired for the inhibitory activity against HIV-1 inte-
grase. Among them, compound 5 c bearing no substitu-
ent at C-8 wasthe most potent (IC
= 20.08 1.9 µM)
50
and showed 6-fold more potency than the styrylquinoline
compound (I a, IC50 = 130.7 8.6 µM) under our assay
condition. Compound 5 f and 6 b, however, bearing
methoxy group and free hydroxy group, respectively, at
the C-8 position of the quinazoline ring, displayed lower
inhibitory activity (IC50 = 31.6 6.1 and 57.1 9.3 µM, re-
spectively) than the parent compound (5 c). Nonethe-
less, compound 5 f and 6 b were more potent than the
corresponding stryrylquinoline compound (I a). Unlike
styrylquinoline compounds (I), in which the hydroxy
group at C-8 wasimportant for the inhibitory activity
against HIV-1 integrase as well as the carboxyl group at
C-7 of the quinoline ring [6], the introduction of a hydroxy
group at C-8 in styrylquinazoline derivatives (II) resulted
in the reduction of the inhibitory activity. Therefore, it
seems that the binding mode of strylquinazoline deriva-
tivesto HIV-1 integrase would be different from that of
styrylquinoline derivatives [6].
HIV-1 integrase inhibitory activity
Styrylquinazoline derivatives( 5 a–f, 6 a, b) were evaluat-
ed for inhibitory activity against 3Ј-processing of HIV-1
integrase [12] and the results are summarized inTable 2.
To compare the data with those of standards, styrylquin-
oline compound (I a) in Figure 1 wasprepared and itsin-
hibitory activities under our assay conditions were in-
serted into the data set. In particular, the IC50 value
(130.7 8.6 µM) of styrylquinoline compound (I a) which
we obtained issignificant higher than the value found by
previousworkers(literature, IC
= 7.4 µM) [6]. It is
50
thought that the difference isat least partly attributable to
the nature of the enzyme and substrate as well as the
overall assay conditions. As shown in Table 2, most of
compounds were largely divided into two classes ac-
cording to the biological activity.For inactive compounds,
in which the catechol moiety is O-acylated (4 c and 4 f)
and/or O-methylated (5 b and 5 e), a complete lack of in-
hibitory activity (IC50 ӷ200 µM) wasobserved, and the
compoundscontaining monoacetoxy ( 4 a) or mono-
In conclusion, the new HIV-1 integrase inhibitors, which
have a quinazoline ring as a scaffold, were synthesized
and evaluated for integrase inhibitory activity. Among
them, the compound (5 c) bearing no substituent at C-8
of the quinazoline subunit was the most potent and
showed 6-fold greater potency than the corresponding
Table 2. HIV-1 integrase inhibitory activities of styrylquinazolines.

280 Lee et al.
Arch. Pharm. Pharm. Med. Chem. 2002, 6, 277–282
methoxybenzaldehyde (135 mg, 2.1 mmol) afforded 4 b
(94 mg, 31 %) asa yellow solid:mp 110–112 °C;IR (KBr) 3008,
1764, 1600, 1506, 1376, 1214, 1018, 750, 470 cm–1; 1H NMR
(CDCl3) δ 9.30 (1 H, s, quinazoline-H4), 8.06 (1 H, d, J =
15.8 Hz, C–CH=CH-phenyl), 7.95 (1 H, d, J = 8.4 Hz, quinaz-
oline-H8), 7.84–7.81 (2 H, m, quinazoline-H5,7), 7.51–7.45
(2 H, m, quinazoline-H6 and phenyl-H6Ј), 7.36 (1 H, s, phenyl-
H2Ј), 7.26 (1 H, d, J = 15.8 Hz, C-CH=CH-phenyl), 6.96 (1 H, d,
J = 8.6 Hz, phenyl-H5Ј), 3.83 (3 H, s, –OCH3), 2.33 (3 H, s,
–OAc); 13C NMR (CDCl3) δ 168.9, 161.2, 160.3, 160.1, 151.8,
150.5, 140.0, 137.4, 134.2, 129.5, 128.0, 126.7, 123.3, 121.6,
112.4, 103.0, 55.9, 20.5.
styrylquinoline compound (I). On the other hand, styryl-
quinazoline (6 b) bearing a free hydroxy group at C-8 of
the quinazoline ring showed less potent activity than the
parent compounds( 5 c and 5 f). Therefore, it isneces-
sary to study further the binding mode of styrylquinaz-
oline derivativesto HIV-1 integrase for the development
of novel antiviral agentsbased upon quinazoline ring as
the scaffold.For more extensive QSAR and the develop-
ment of more potent compound, we are undertaking the
synthesis of styrylquinazolines containing a carboxyl
group at the C-7 position of the quinazoline ring.
(E)-2-[2-(3,4-Diacetoxyphenyl)ethenyl]quinazoline (4 c)
A similar procedure to that for the preparation of 4 a using 2-
methylquinazoline (2 a) (70 mg, 0.4 mmol) and 3,4-dihydroxy-
benzaldehyde (221 mg, 1.6 mmol) afforded 4 c (157 mg, 45 %)
asa yellow solid: mp 120–123 °C; IR (KBr) 2930, 2674, 1758,
1376, 1216, 1108, 1014, 754, 582 cm–1;1H NMR (CDCl3) δ 9.37
(1 H, s, quinazoline-H4), 8.12 (1 H, d, J = 15.9 Hz, C-CH=CH-
phenyl), 8.01 (1 H, d, J = 8.0 Hz, quinazoline-H8), 7.92–7.87
(2 H, m, quinazoline-H5,7), 7.63–7.60 (1 H, m, quinazoline-
H6), 7.58–7.56 (1 H, dd, J = 8.4, 2.0 Hz, phenyl-H2Ј), 7.50 (1 H,
d, J = 2.0 Hz, phenyl-H6Ј), 7.39 (1 H, d, J = 15.9 Hz, C-CH=CH-
phenyl), 7.25 (1 H, d, J = 8.4 Hz, phenyl-H5Ј), 2.33 (3 H, s,
-OAc), 2.31 (3 H, s, -OAc); 13C NMR (CDCl3) δ 168.6, 161.3,
160.7, 150.9, 142.9, 142.8, 137.0, 135.6, 134.7, 129.5, 128.6,
127.8, 127.6, 126.2, 124.2, 123.8, 122.7, 21.1.
Acknowledgements
We are grateful to Ministry of Science and Technology,
Korea for the support of this work (2N23260).
Experimental
Chemistry
General
1H and 13C NMR spectra were recorded on a Gemini Varian-
300 (300 and 75 MHz, respectively). Mass spectra (EI) were
determined on HP GC 5972 and HP MS 5988A system at
70 eV. Melting points(mp) were determined on a Thomas-
Hoover capillary melting apparatusand are uncorrected. Infra-
red (IR) spectra were recorded on Perkin Elmer 16F-PC FT-IR
and MIDAC 101025 using a potassium bromide pellet. Analyti-
cal thin layer chromatography (TLC) wascarried out on pre-
coated silica gel (E. Merck Kiesegel 60F254 layer thickness
0.25 mm). Flash column chromatography was performed with
Merck Kiesegel 60 Art 9385 (230–400 mesh).All solvents used
were purified according to standard procedures.
(E)-8-Methoxy-2-[2-(4-acetoxyphenyl)ethenyl]quinazoline
(4 d)
A similar procedure to that for the preparation of 4 a using 2-
methyl-8-methoxyquinazoline (2 b) (120 mg, 0.7 mmol) and
4-hydroxybenzaldehyde (337 mg, 2.8 mmol) afforded 4 d
(77 mg, 47 %) asa yellow solid:mp 149–152 °C;IR (KBr) 3450,
2932, 2364, 1760, 1558 cm–1; 1H NMR (CDCl3) δ 9.35 (1 H, s,
quinazoline-H4), 8.15 (1 H, d, J = 16.0 Hz, C-CH=CH-phenyl),
7.69 (2 H, d, J = 8.6 Hz, phenyl-H2Ј,6Ј), 7.55–7.46 (3 H, m,
quinazoline-H5,7 and C-CH=CH-phenyl), 7.25 (1 H, dd, J = 7.3
Hz, 1.5 Hz, quinazoline-H6), 7.15 (1 H, d, J = 8.6 Hz,
Perkin condensation between quinazoline and various aro-
matic aldehydes
phenyl-H3Ј,5Ј), 4.12 (3 H, s, -OCH3), 2.32 (3 H, s, -OAc); 13
C
NMR (CDCl3) δ 169.7, 161.0, 160.4, 155.0, 151.5, 137.5,
134.5, 129.1, 128.9, 127.8, 124.6, 122.3, 119.1, 112.4, 56.6,
21.5.
(E)-2-[2-(4-Acetoxyphenyl)ethenyl]quinazoline (4 a)
To a solution of 2-methylquinazoline (2 a) (100 mg, 0.7 mmol) in
3 mL of acetic anhydride wasadded 4-hydroxybenzaldehyde
(341 mg, 2.1 mmol).The reaction mixture washeated under re-
flux for 3 days. After cooling to rt, the mixture was poured into a
mixture of saturated NaHCO3 solution and CH2Cl2.The organic
layer wasseparated and the aqueouslayer wasextracted with
(E)-8-Methoxy-2-[2-(3-acetoxy-4-methoxyphenyl)ethenyl]-
quinazoline (4 e)
The similar procedure for the preparation of 4 a using 2-methyl-
8-methoxyquinazoline (2 b) (250 mg, 1.4 mmol) and 3-hydroxy-
4-methoxybenzaldehyde (656 mg, 4.3 mmol) afforded 4 e
(372 mg, 73 %) asa bright yellow amorphousoslid: mp
60–65 °C; IR (KBr) 2930, 2366, 1766, 1610, 1512, 1268, 1020
CH2Cl2.The combined organic extractswasdried over MgSO
,
4
concentrated, and purified by flash column chromatography
(EtOAc : CH2Cl2 = 1 : 5) to give 4 a (54 mg, 27 %) asa yellow
solid: mp 108–112 °C; IR (KBr) 2934, 2552, 1756, 1686, 1426,
1372, 1198, 914, 758, 504 cm–1;1H NMR (CDCl3) δ 9.42 (1 H, s,
quinazoline-H4), 8.18–8.12 (3 H, m, C–CH=CH-phenyl, phe-
nyl-H2Ј6Ј), 8.06 (1 H, d, J = 8.8 Hz, quinazoline-H8), 7.92–7.88
(1 H, m, quinazoline-H7), 7.71 (1 H, d, J = 8.6 Hz, quinazoline-
H5), 7.62–7.57 (1 H, m, quinazoline-H6), 7.45 (1 H, d, J = 16.0
Hz, C–CH=CH-phenyl), 7.23–7.19 (2 H, d, J = 8.7 Hz, phenyl-
H3Ј,5Ј), 2.33 (3 H, s, –OAc); 13C NMR (CDCl3) δ 170.0, 160.9,
160.6, 138.2, 135.0, 134.8, 132.1, 129.3, 129.0, 128.1, 127.6,
122.8, 122.0, 121.8, 21.6.
1
cm–1; H NMR (CDCl3) δ 9.20 (1 H, s, quinazoline-H4), 7.97
(1 H, d, J = 15.9 Hz, C-CH=CH-phenyl), 7.41–7.32 (5 H, m,
quinazoline-H5,6,7, phenyl-H2Ј and C-CH=CH-phenyl), 7.09
(1 H, d, J = 8.4 Hz, phenyl-H6Ј), 6.89 (1 H, d, J = 8.4 Hz, phe-
nyl-H5Ј), 3.98 (3 H, s, -OCH3), 3.75 (3 H, s, -OCH3), 2.23 (3 H, s,
-OAc); 13C NMR (CDCl3) δ 169.4, 160.9, 160.3, 154.8, 152.1,
142.9, 140.3, 137.4, 129.9, 127.7, 127.4, 127.1, 124.4, 122.1,
119.1, 112.8, 112.3, 56.6, 56.3, 21.1.
(E)-8-Methoxy-2-[2-(3,4-diacetoxyphenyl)ethenyl]quinazoline
(E)-2-[2-(3-Acetoxy-4-methoxyphenyl)ethenyl]quinazoline
(4 f)
(4 b)
A similar procedure to that for the preparation of 4 a using 2-
methyl-8-methoxyquinazoline (2 b) (70 mg, 0.4 mmol) and 3,4-
dihydroxybenzaldehyde (221 mg, 1.6 mmol) afforded 4 f
A similar procedure to that for the preparation of 4 a using 2-
methylquinazoline (2 a) (100 mg, 0.7 mmol) and 3-hydroxy-4-

Arch. Pharm. Pharm. Med. Chem. 2002, 6, 277–282
Styrylquinazoline Derivatives281
(E)-8-Methoxy-2-[2-(4-hydroxyphenyl)ethenyl]quinazoline
(5 d)
(103 mg, 47 %) asa yellow solid: mp 138–141 °C; IR (KBr)
2947, 1760, 1560, 1378, 1258, 1214, 1012, 898, 762 cm–1; 1H
NMR (CDCl3) δ 9.34 (1 H, s, quinazoline-H4), 8.10 (1 H, d, J =
16.0 Hz, C-CH=CH-phenyl), 7.56–7.44 (5 H, m, quinazoline-
H5,6,7 and phenyl-H2Ј, C-CH=CH-phenyl), 7.24 (2 H, m, phe-
nyl-H5Ј,6Ј), 4.11 (3 H, s, -OCH3), 2.33 (3 H, s, -OAc), 2.31 (3 H,
s , -OAc);13C NMR (CDCl3) δ 168.5, 160.7, 160.4, 155.0, 142.8,
136.7, 135.8, 129.8, 127.9, 126.1, 124.6, 124.1, 122.7, 119.1,
112.4, 56.6, 21.0.
A similar procedure to that for the preparation of 5 c using com-
pound 4 d (254 mg, 0.7 mmol) afforded 5 d (37 mg, 17 %) asa
yellow solid: mp 170–174 °C; IR (KBr) 2928, 1604, 1560, 1514
1
cm–1; H NMR (CDCl3) δ 9.28 (1 H, s, quinazoline-H4), 8.02
(1 H, d, J = 16.0 Hz, C-CH=CH-phenyl), 7.57–7.34 (4 H, m,
quinazoline-H5,6,7 and C-CH=CH-phenyl), 7.28–7.17 (2 H, m,
phenyl-H2Ј,6Ј), 6.82 (2 H, d, J = 8.5 Hz, phenyl-H3Ј, 5Ј), 4.05
(3 H, s, -OCH3); 13C NMR (CDCl3) δ 163.9, 163.2, 161.6, 157.3,
145.0, 142.0, 133.4, 132.4, 130.9, 130.6, 127.0, 121.9, 118.8,
115.6, 58.9.
General procedure for deacetylation of acetylated compounds
(4 a, 4 b, and 4 e)
(E)-2-[2-(4-Hydroxyphenyl)ethenyl]quinazoline (5 a)
(E)-2-[2-(4-Acetoxyphenyl)ethenyl]quinazoline (4 a) (46 mg,
0.2 mmol) was dissolved in 2 mL of 0.5 N NaOH, and the result-
ing solution was heated under reflux for 10 min. After cooling to
rt, the reaction mixture wasextracted with CH 2Cl2.The organic
phase was dried over MgSO4 and concentrated to give 5 a
(22 mg, 56 %) asa white solid: mp 227–230 °C (dec); IR (KBr)
3012, 2362, 1598, 1454, 1402, 1254, 1172, 974, 828, 762, 642
cm–1; 1H NMR (CD3OD) δ 9.42 (1 H, s, quinazoline-H4), 8.11
(1 H, d, J = 15.9 Hz, C-CH=CH-phe2nyl), 8.03 (1 H, d, J = 8.1
Hz, quinazoline-H8), 7.94–7.92 (2 H, m, quinazoline-H5,7),
7.66–7.61 (1 H, m, quinazoline-H6), 7.56 (2 H, d, J = 8.6 Hz,
phenyl-H2Ј,6Ј), 7.19 (1 H, d, J = 15.9 Hz, C-CH=CH-phenyl),
6.86 (2 H, d, J = 8.6 Hz, phenyl-H3Ј,5Ј); 13C NMR (CD3OD) δ
149.7, 149.3, 148.8, 138.4, 127.8, 123.1, 117.8, 117.4, 115.8,
115.3, 111.8, 111.5, 104.0, 103.3.
(E)-8-Methoxy-2-[2-(3-hydroxy-4-methoxyphenyl)ethenyl]-
quinazoline (5 e)
A similar procedure to that for the preparation of 5 a using com-
pound 4 e (50 mg, 0.2 mmol) afforded 5 e (30 mg, 65 %) asa
bright yellow solid: mp 197–200 °C (dec.); IR (KBr) 3456, 1624,
1572, 1466, 1344, 986, 762,474 cm–1;1H NMR (CD3OD) δ 9.30
(1 H, s, quinazoline-H4), 8.04 (1 H, d, J = 15.9 Hz, C-CH=CH-
phenyl), 7.54–7.45 (2 H, m, quinazoline-H5,7), 7.32–7.25 (2 H,
m, C-CH=CH-phenyl and quinazoline-H6), 7.11 (1 H, d, J =
1.6 Hz, phenyl-H2Ј), 6.89 (1 H, dd, J = 8.1, 1.6 Hz, phenyl-H6Ј),
6.81 (1 H, d, J = 8.2 Hz, phenyl-H5Ј), 4.30 (3 H, s, -OCH3), 4.10
(3 H, s, -OCH3); 13C NMR (CD3OD) δ 162.9, 161.8, 158.0,
155.7, 154.9, 143.7, 142.7, 131.2, 128.8, 125.5, 124.3, 120.4,
117.7, 117.1, 114.3, 112.0, 57.0, 56.3.
(E)-2-[2-(3-Hydroxy-4-methoxyphenyl)ethenyl]quinazoline
(5 b)
(E)-8-Methoxy-2-[2-(3,4-dihydroxyphenyl)ethenyl]quinazoline
(5 f)
A similar procedure to that for the preparation of 5 a using com-
pound 4 b (50 mg, 0.2 mmol) afforded 5 b (42 mg, 89 %) asa
yellow solid: mp 107–110 °C; IR (KBr) 3142, 1608, 1506, 1398,
A similar procedure to that for the preparation of 5 c using com-
pound 4 f (80 mg, 0.3 mmol) afforded 5 f (80 mg, 63 %) asa yel-
low solid: mp 204–208 °C; IR (KBr) 3198, 2930, 1562,
1518 cm–1;1H NMR (DMSO-d6) δ 9.47 (1 H, s, quinazoline-H4),
7.93 (1 H, d, J = 15.9 Hz, C-CH=CH-phenyl), 7.61–7.56 (2 H, m,
quinazoline-H5,7), 7.41 (1 H, d, J = 7.3 Hz, quinazoline-H6),
7.13–7.04 (3 H, m, phenyl-H2Ј,6Ј, C-CH=CH-phenyl), 6.81
(1 H, d, J = 8.1 Hz, phenyl-H5Ј), 4.11 (3 H, s, -OCH3); 13C NMR
(DMSO-d6) δ 160.6, 160.4, 154.5, 147.5, 145.9, 138.4, 127.7,
124.9, 124.0, 120.7, 119.1, 116.2, 114.5, 113.3, 55.2.
1272, 1030, 978, 758 cm–1; H NMR (CDCl3) δ 9.35 (1 H, s,
1
quinazoline-H4), 8.09 (1 H, d, J = 15.9 Hz, C-CH=CH-phenyl),
8.00 (1 H, d, J = 8.8 Hz, quinazoline-H8), 7.90–7.85 (2 H, m,
quinazoline-H5,7), 7.59–7.55 (1 H, m, quinazoline-H6), 7.29
(1 H, s, phenyl-H2Ј), 7.24–7.16 (1 H, m, C-CH=CH-phenyl and
phenyl-H6Ј), 6.89 (1 H, d, J = 8.3 Hz, phenyl-H5Ј), 3.91 (3 H, s,
-OCH3); 13C NMR (CDCl3) δ 161.8, 159.4, 139.8, 137.7, 135.6,
133.5, 129.5, 128.4, 127.4, 126.3, 125.5, 122.3, 120.2, 114.4,
112.1, 110.0, 55.4.
General procedure for deacetylation of acetylated compounds
(4 c, 4 d, and 4 f)
General procedure for demethylation of compounds (5 d and
5 f)
(E)-2-[2-(3,4-Dihydroxyphenyl)ethenyl]quinazoline (5 c)
(E)-8-Hydroxy-2-[2-(4-hydroxyphenyl)ethenyl]quinazoline
(E)-2-[2-(3,4-Diacetoxyphenyl)ethenyl]quinazoline 4 c
(254 mg, 0.7 mmol) was dissolved in 1.5 mL of pyridine, and
the resulting solution was heated under reflux for 4 h. 1 mL of
water wasthen added to the mixture and the reaction mixture
wasfurther heated under reflux for 1 h. After cooling to rt, the
reaction mixture wasextracted with CH 2Cl2.The organic phase
wasdried over Na 2SO4 and concentrated to leave a solid,
which wasrecrystallized from EtOAc and ether to provide 5 c
(209 mg, 60 %) asa yellow solid: mp 204–208 °C (dec.); IR
(KBr) 3545, 3066, 2378, 1608, 1406 cm–1; 1H NMR (acetone-
d6) δ 9.56 (1 H, s, quinazoline-H4), 8.17 (1 H, d, J = 15.9 Hz, C-
CH=CH-phenyl), 8.16 (1 H, d, J = 8.0 Hz, quinazoline-H8),
8.06–7.99 (2 H, m, quinazoline-H5,7), 7.75–7.70 (1 H, m,
quinazoline-H6), 7.36 (1 H, d, J = 2.0 Hz, phenyl-H2Ј), 7.28
(1 H, d, J = 15.9 Hz, C-CH=CH-phenyl), 7.23 (1 H, dd, J =
8.2 Hz, 2.0 Hz, phenyl-H6Ј), 7.00 (1 H, d, J = 8.2 Hz,
phenyl-H5Ј); 13C NMR (acetone-d6) δ 162.9, 161.6, 151.9,
148.1, 146.7, 139.7, 135.4, 129.9, 129.1, 128.8, 128.1, 126.5,
124.6, 122.1, 116.8, 115.1.
(6 a)
To a mixture of (E)-8-methoxy-2-[2-(4-hydroxyphenyl)ethenyl]-
quinazoline (5 d) (55 mg, 0.2 mmol) in 8 mL of CH2Cl2 wasadd-
ed BBr3 (1 M solution in n-hexane, 1.7 mL, 1.7 mmol) under N2
atmosphere.The reaction mixture was stirred at rt for 18 h and
quenched with water.The organic layer wasseparated, washed
with saturated NaHCO3 solution and brine successively, and
dried over MgSO4.After concentration, the residue was purified
by flash column chromatography (EtOAc : n-hexane = 3 : 2) to
afford 6 a (11 mg, 21 %) asa brown-yellow oslid: mp
197–200 °C (dec.); IR (KBr) 3148, 2930, 1602, 1560 cm–1; 1H
NMR (CD3OD) δ 9.22 (1 H, s, quinazoline-H4), 8.01 (1 H, d, J =
16.0 Hz, C-CH=CH-phenyl), 7.46 (2 H, d, J = 8.5 Hz,
phenyl-H2Ј,6Ј), 7.37 (2 H, m, quinazoline-H5,7), 7.21 (1 H, m,
quiazoline-H6), 7.17 (1 H, d, J =16.0 Hz, C-CH=CH-phenyl),
6.76 (1 H, d, J = 8.5 Hz, phenyl-H3Ј,5Ј); 13C NMR (CD3OD) δ
161.7, 160.3, 154.0, 142.8, 140.2, 130.8, 129.6, 129.5, 125.5,
119.2, 117.6, 117.2, 116.2.

282 Lee et al.
Arch. Pharm. Pharm. Med. Chem. 2002, 6, 277–282
(E)-8-Hydroxy-2-[2-(3,4-dihydroxyphenyl)ethenyl]quinazoline
(6 b)
References and notes
[1] F. Barre-Sinoussi, J. C. Chermann, F. Rey, M. T. Nugeyre,
S.Chemaret, J.Gruest, C.Dauguet, C.Axler-Blin, F.Byun-
Vezinet, C. Rouzioux, W. Rozenbaum, L. Montagnier, Sci-
ence, 1983, 220, 868–871.
A similar procedure to that for the preparation of 6 a using com-
pound 5 f (117 mg, 0.3 mmol) afforded 6 b (22 mg, 24 %) asa
yellow solid: mp 160 °C (dec.); IR (KBr) 3390, 2364, 1562,
1470, 1392, 1278, 762 cm–1; 1H NMR (CD3OD) δ 9.22 (1 H, s,
quinazoline-H4), 7.95 (1 H, d, J = 15.9 Hz, C-CH=CH-phenyl),
7.36–7.34 (2 H, m, quinazoline-H5,7), 7.19 (1 H, t, J = 4.7 Hz,
quinazoline-H6), 7.10 (1 H, d, J = 15.9 Hz, C-CH=CH-phenyl),
7.07 (1 H, d, J = 1.8 Hz, phenyl-H2Ј), 6.93 (1 H, dd, J = 8.2 Hz,
J = 1.7 Hz, phenyl-H6Ј), 6.72 (1 H, d, J = 8.2 Hz, phenyl-H5Ј);
13C NMR (CD3OD) δ 160.3, 160.28, 152.7, 147.3, 145.7, 141.3,
139.2, 128.7, 128.0, 124.1, 123.9, 121.0, 117.7, 116.1, 115.5,
113.7.
[2] P. O. Brown, Curr. Top. Microbiol. Immunol. 1990, 157,
19–48.
[3] Y. Pommier, N. Neamati, Adv. Virus Res. 1999, 52,
427–458.
[4] A. Mazumder, N. Neamati, J.-P. Sommadossi, G. Gosse-
lin, R. F. Schinazi, J.-L. Imbach,Y. Pommier, Mol. Pharma-
col. 1996, 49, 621–628; A. Mazumder, N. Neamati, J. O.
Ojwang, S.Sunder, R.F.Rando,Y.Pommier, Biochemistry
1996, 35, 13762–13771; J. O. Ojwang, R. W. Buckheit, Y.
Pommier, A. Mazumder, K. de Vreese, J. A. Este, D.
Reymen, L.A.Pallansch, C.Lackman-Smith,T.L.Wallace,
Antimicrob. Agents Chemother. 1995, 39, 2426–2435; J.
K. Buolamwini, H. Assefa, J. Med. Chem. 2002, 45,
841–852; Aronex Pharmaceutical, Woodland, TX and
New York Hospital.
Biological test
HIV-1 integrase
Recombinant human immunodeficiency virustype 1 (HIV-1) in-
tegrase was expressed in Escherichia coli and purified using a
nickel-chelated column in an one-step manner.Aliquots of HIV-
1 integrase of 0.5 mg/mL as stock solutions were stored at
–70 °C until used.
[5] M. Thomas, L. Brady, TIBTECH 1997, 15, 167–172.
Oligonucleotide substrates
Two 20-mer oligonucleotides whose sequences resemble the
end of U5-LTR were obtained from Korea Biotech. Inc.: K16
(U5-LTR, +strand), 5Ј-TGTGGAAAATCTCTAGCAGT-3Ј; K17
(U5-LTR, -strand), 5Ј-ACTGCTAGA-GATTTTCCACA-3Ј. The
oligonucleotideswere purified with 20 % polyacrylamide gel
before use. In order to construct oligonucleotide substrate, oli-
gonucleotide K16 of 30 pmol waslabeled at the 5 Ј end using
[γ-32P]-ATP of 250 µCi (3,000 Ci/mmol; 1 Ci = 37 GBq; Amer-
sham) and T4 polynucleotide kinase (T4 PNK, New England
Biolabs) of 10 units in 40 µL of reaction buffer (70 mM Tris-HCl
[pH 7.6], 10 mM MgCl2, 5 mM dithiothreitol) at 37 °C for 15 min.
The labeling reaction wassubjected to 10 mM EDTA, and heat-
ed to 85 °C for 15 min to inactivate T4 PNK. After addition of
complementary oligonucleotide K17 of 30 pmol, the reaction
mixture wasboiled for 3 min and cooled down slowly. Labeled
substrate was separated from unincorporated nucleotide by
passage through a Biospin 6 (Bio-Rad).
[6] K. Mekouar, J.-F. Mouscadet, D. Desmaële, F. Subra, H.
Leh, D. Savouré, C. Auclair, J. d’Angelo, J. Med. Chem.
1998, 41, 2846–2857; F. Zouhiri, J. Mouscadet, K.
Mekouar, D.Desmaële, D.Savouré, H.Leh, F.Subra, M.L.
Bret, C. Auclair, J. d’Angelo, J. Med. Chem. 2000, 43,
1533–1540.
[7] A.R.Katritzky, C.W.Rees, E.F.V.Scriven, Comprehensive
Heterocyclic Chemistry II, Vol. 6 (Ed.: A. J. Boulton),
Pergamon, New York, 1996, chapter 2.
[8] D.J. Brown, Fused Pyrimidines: Part I Quinazolines.Inter-
science Publisher, New York, 1967; W. L. F. Armarego, J.
Chem. Soc. 1962, 561–572; E. S. Hand, D. C. Baker, Can.
J. Chem. 1984, 62, 2570–2577; A. B. Sen, S. K. Gupta, J.
Indian. Chem. Soc. 1962, 39, 368–372.
[9] H. Irving, A.R. Pinnington, J. Chem. Soc. 1954,
3782–3785.
HIV-1 integrase reaction
[10] R. L. Lafemina, P. L. Graham, K. LeGrow, J. C. Hastings, A.
Wolfe, S. D.Young, E. A. Emini, D. J. Hazuda, Antimicrob.
Agents Chemother. 1995, 39, 320–324.
A standard reaction assay of the endonucleolytic activity was
carried out in the presence of potential inhibitor containing
0.1 pmol of duplex oligonucleotide substrate and 15 pmol of
HIV-1 integrase in 15 mM Tris-HCl (pH 7.4), 100 mM NaCl,
1 mM MnCl2, 2 mM 2-mercaptoethanol, 2.5 mM CHAPS,
0.1 mM EDTA, 0.1 mM PMSF, 1 % glycerol, and 10 mM imid-
azole in a total volume of 10 µL. Inhibitorsor drugswere dis-
solved in 100 % DMSO and added to the reaction to be 5 %
DMSO in the final volume.Reaction mixtureswere incubated at
33 °C for 90 min and stopped by addition of 4 µL of 95 % form-
amide, 20 mM EDTA, 0.05 % bromophenol blue, 0.05 % xylene
cyanol FF. The reactionswere heated to 90 °C for 3 min and
electrophoresed on a 20 % denaturing polyacrylamide gel. Re-
action productswere visualized by autoradiography of the wet
gel. IC50 wascalculated by scanning bandson Kodak-5 film
(Image Master VDS, Pharmacia Biotech).
[11] A. S. Kende, J. P. Rizzi, Tetrahedron Lett. 1981, 22,
1779–1782.
[12] The enzyme inhibition assay of compounds against HIV-1
integrase was carried out as described previously: J.-W.
Oh, C.-G. Shin, Mol. Cells 1996, 6, 96–100.