Synthesis of novel diarylpyrimidine analogues and their antiviral activity against human immunodeficiency virus type 1

Article abstract of DOI:10.1021/jm040838n

This paper reports the synthesis and the antiviral properties of new diarylpyrimidine (DAPY) compounds as nonnucleoside reverse transcriptase inhibitors (NNRTIs). The synthesis program around this new DAPY series was further optimized to produce compounds displaying improved activity against a panel of eight clinically relevant single and double mutant strains of human immunodeficiency virus type 1 (HIV-1).

Full text of DOI:10.1021/jm040838n

2072
J. Med. Chem. 2005, 48, 2072-2079
Synthesis of Novel Diarylpyrimidine Analogues and Their Antiviral Activity
against Human Immunodeficiency Virus Type 1
Jerome Guillemont,*,† Elisabeth Pasquier,† Patrice Palandjian,† Daniel Vernier,† Sandrine Gaurrand,†
Paul J. Lewi,‡ Jan Heeres,‡ Marc R. de Jonge,‡ Lucien M. H. Koymans,‡ Frits F. D. Daeyaert,‡
Maarten H. Vinkers,‡ Eddy Arnold,§ Kalyan Das,§ Rudi Pauwels,| Koen Andries,| Marie-Pierre de Be´thune,|
Eva Bettens,| Kurt Hertogs,| Piet Wigerinck,| Philip Timmerman, and Paul A. J. Janssen‡,∇
Medicinal Chemistry Department, Johnson & Johnson Pharmaceutical Research and Development, Campus de Maigremont
BP615, Val de Reuil, France, Center for Molecular Design, Janssen Pharmaceutica NV, Vosselaar, Belgium, Tibotec,
General De Wittelaan L11B3, B-2800 Mechelen, Belgium, Pharmacokinetic Department, Johnson & Johnson Pharmaceutical
Research and Development, Tumhoutseweg 30 B-2340 Beerse, Belgium, and Center for Advanced Biotechnology and Medicine,
Chemistry Department, Rutgers University, 679 Hoes Lane, Piscataway, New Jersey 08854
Received May 24, 2004
This paper reports the synthesis and the antiviral properties of new diarylpyrimidine (DAPY)
compounds as nonnucleoside reverse transcriptase inhibitors (NNRTIs). The synthesis program
around this new DAPY series was further optimized to produce compounds displaying improved
activity against a panel of eight clinically relevant single and double mutant strains of human
immunodeficiency virus type 1 (HIV-1).
Introduction
The reverse transcriptase (RT) of the human immuno-
deficiency virus type 1 (HIV-1) is a key enzyme, which
plays an essential and multifunctional role in the
replication cycle. This enzyme is responsible for the
conversion of a single-stranded RNA genome of HIV-1
into a double-stranded DNA chain that subsequently
is incorporated into the DNA of the infected host cell.
Nonnucleoside inhibitors of HIV-1 reverse transcriptase1
(NNRTIs) inhibit the enzyme by occupation of an
induced allosteric binding site very close to the active
site.2 The emergence of resistant HIV viral strains is a
limitation for all therapeutic classes.3,4 The emerging
cross-resistance among the approved drugs5-7 (Figure
1) has particularly limited the use of the NNRTIs class.
In addition, there is the need to develop new NNRTIs
with enhanced side effect profiles and improved char-
acteristics influencing drug compliance.
Figure 1. NNRTIs approved by the FDA.
TMC125,8,9 4a (Figure 2), an early diarylpyrimidine
(DAPY) compound, was the first NNRTI demonstrating
a beneficial effect on HIV-infected patients with NNRTI-
resistant viruses. TMC1259 is currently in phase IIB
clinical trials.
However, criteria like drug compliance, adverse ef-
fects, and cross-resistance restrict the development of
new drug candidates. As a result of these factors, there
is a real medical need to develop new NNRTIs, which
do not give rise to cross-resistance and are effective
against clinically relevant mutant strains.
Figure 2. TMC125 (R165335) 4a and TMC120 (R147681) 4b.
modeling studies by use of available crystallographic
structures of RT complexes with NNRTIs and was
synthetically straightforward.10 This modeling approach
indicated that molecular interaction energy with certain
conserved key amino acids of HIV-RT could be improved
by replacement of the NH2 moiety with a hydrogen atom
on the central pyrimidine ring, which was subsequently
confirmed by in vitro HIV activity.10 Furthermore,
comparison of the binding energies of compounds 4b
(dapivirine) and 5 indicated that improving the interac-
tion between the para substituent on the trisubstituted
phenyl ring and the conserved W229 region within the
RT enzyme could give rise to novel variations of DAPY
analogues with improved activity against wt LAI virus
The DAPY compounds were developed from the DATA
series.10 The modifications were inferred from molecular
* To whom correspondence should be addressed: phone +33 2 32
61 74 58; fax +33 2 32 61 72 98; e-mail jguillem@prdfr.jnj.com.
† Johnson & Johnson Pharmaceutical Research and Development,
France.
‡ Janssen Pharmaceutica NV.
| Tibotec.
Johnson & Johnson Pharmaceutical Research and Development,
Belgium.
§ Rutgers.
∇ This paper is dedicated to the memory of Dr. Paul Adriaan Jan
Janssen (1926-2003).
10.1021/jm040838n CCC: $30.25 © 2005 American Chemical Society
Published on Web 11/18/2004

DAPY Analogues: Antiviral Activity against HIV-1
Journal of Medicinal Chemistry, 2005, Vol. 48, No. 6 2073
Ester 10 was prepared by mixing ethyl 4-amino-3,5-
dimethylbenzoate 8 and chloropyrimidine derivative 910
at 150 °C for 2 h. Thereupon carboxylic ester 10 was
reduced to alcohol 13 with lithium aluminum hydride
(LiAlH4) without affecting the cyano function present
on the scaffold. Manganese dioxide (MnO2) oxidation of
alcohol 13 furnished the corresponding aldehyde 14.
Bromine derivative 12 was synthesized according to the
same method as described for compound 10.
Two methods have been used to gain access to these
new DAPY compounds. Two basic methods were uti-
lized, either Wadsworth-Emmons13 or Wittig reaction
conditions starting from aldehyde 14 (Scheme 2) or
alternatively making use of the Heck reaction14 starting
from bromine derivative 12 as an intermediate (Scheme
3).
Figure 3. Introduction of G spacer group between the aryl
ring and the cyano group.
First of all, we investigated the Wittig and
Wadsworth-Emmons reactions. Aldehyde 14 was con-
densed with either cyanomethyl triphenylphosphonium
chloride 15a (R1dH) (Scheme 2) or diethyl (1-cyano-
ethyl) phosphonate 15b (R1 ) Me) by use of t-BuOK in
tetrahydrofuran (THF) as condensing agent to give the
corresponding vinylcyanides 6, which were isolated with
the following stereoselectivity: 6a (E)/6b (Z) ) 80/20
and 6c (E)/6d (Z) ) 44/56. Hydrogenation of vinyl
cyanides 6 with palladium on activated carbon (Pd/C
10%) as a catalyst in methanol afforded the expected
reduced compounds 7. Target compound 6e (ratio E/Z
) 12/88) could be prepared by treatment of aldehyde
14 in THF with fumaronitrile 16a in the presence of
tri-n-butylphosphine.15
As a second method we used the Heck reaction
(Scheme 3) in order to introduce the acrylonitrile group
(either substituted or not) directly. Compounds 6a and
6b (ratio E/Z ) 90/10) have been obtained via this
method after treatment of aryl bromide 12 with acryl-
onitrile 16b in the presence of triethylamine and pal-
ladium acetate as a catalyst in acetonitrile. Under the
same conditions, compound 6f (E) has been prepared
from the crotononitrile reagent 16c. Compound 7c was
and probably should have beneficial effects on activity
against mutants. This observation also lead to the
introduction of a spacer group G between the cyano
group in 4-position and the trisubstituted phenyl ring
(Figure 3). Subsequently, DAPY compounds 6 and 7
were based both on structure-activity relationships
(SAR)10 and on molecular modeling studies11 of the 2-(4-
cyanoanilino)-4-(4-cyano-2,6-dimethylanilino)pyrimi-
dine 5 family by use of X-ray crystal structures studies
of the RT-R147681 (4b, dapivirine) complex.12 The
resulting compounds were tested for in vitro activities
against the wild type (WT) and against a panel of
clinically relevant single and double NNRTI mutant
strains of HIV activity.
These efforts resulted in the synthesis of a new class
of DAPY compounds, 6 and 7, which combine very high
potency against the wild type and a panel of single and
double mutant strains of HIV-1 with minimal in vitro
toxicity.
Chemistry
Newly designed compounds 6 and 7 were obtained
from key intermediates 12 and 14 as depicted in Scheme
1. Aldehyde 14 was easily synthesized in three steps.
Scheme 1a
a Conditions: (a) 150 °C, 2 h; (b) LiAlH4, THF, 0 °C to room temperature; (c) MnO2, CH2Cl2, room temperature, 20 h; (d) 150 °C, 1 h.

2074 Journal of Medicinal Chemistry, 2005, Vol. 48, No. 6
Guillemont et al.
Scheme 2a
a Conditions: (a) tBuOK, THF, cyanomethyl triphenylphosphonium chloride 15a, room temperature; (b) tBuOK, THF, diethyl (1-
cyanoethyl)phosphonate 15b, room temperature; (c) fumaronitrile 16a, P(Bu3), THF, reflux; (d) Pd/C, MeOH, H2 (3 bar).
Scheme 3a
a Conditions: (a) CH2dCH-CN (16b) or Me-CHdCH-CN (16c) or CH2dCH-CH2-CN (16d), Pd(OAc)2, P(o-tol)3, Et3N, CH3CN, 150
°C; (b) Pd/C, MeOH, H2 (3 bar).
Scheme 4a
a Conditions: (a) LDA, THF, CH3CN, -70 °C, 2 h; (b) Jones’ reagent, acetone, 0 °C, 2 h; (c) POCl3, 80 °C, 24 h.
Scheme 5a
prepared by catalytic hydrogenation with palladium on
activated carbon (Pd/C 10%) in methanol from com-
pounds 6f. E isomer compound 6g could specifically be
prepared via Heck reaction conditions starting from
allylcyanide 16d and 12. The synthesis of â-chloroacryl-
onitrile compound 6h is depicted in Scheme 4. Aldehyde
14 was reacted with the lithium salt of acetonitrile to
give the hydroxycyano intermediate 17 (indicated in
Scheme 4). Oxidation of 17 with Jones reagent in
acetone led to ketone 18. Subsequently this ketone was
then converted with phosphorus oxychloride to give the
desired compound 6h (ratio E/Z ) 70/30).
The synthesis of 2-furyl heterocycle derivatives 20,
21, and 7d is described in Scheme 5. The first step
involved a modified in situ Suzuki cross-coupling reac-
tion the coupling of 4-bromofurylcarboxaldehyde 19 with
the bromo derivative 12 by use of the tetraalkoxydiboron
derivative 22,16 which is thermally stable and can be
easily handled in air. Moreover it has been shown to be
a useful boron nucleophile for the cross-coupling reac-
tion with heterocycles halides containing base-sensitive
functionalities.17
a Conditions: (a) Bis(pinacolato)diboron 22, Pd(PPh3)4, K2CO3,
toluene, EtOH, H2O, reflux, 48 h; (b) NH2OH, HCl, 23, EtOH,
NaOH (5 N), 50 °C, 2 h; (c) CDI, THF, reflux, 9 h.
Furylaldehyde derivative 20 was transformed with
hydroxylamine chloride 23 in ethanol into oxime 21 in
the presence of sodium hydroxide in ethanol. Subse-
quently cyano derivative 7d was then obtained from 23

DAPY Analogues: Antiviral Activity against HIV-1
Journal of Medicinal Chemistry, 2005, Vol. 48, No. 6 2075
Scheme 6a
activity against the LAI virus and the panel of mutant
strains with a good selectivity index (SI).
The new series of DAPY compounds (6-7) was much
more potent against wild-type virus than nevirapine and
delavirdine and showed a comparable efficacy to the
cyano derivative 5 and efavirenz against this virus type.
Replacement of the cyano substituent (5) by an
acrylonitrile group 6a (G ) vinyl, pure E-isomer) led to
a very potent compound highly active against single and
double mutant strains, resulting in EC50 values equal
or lower than 2 nΜ against LAI virus and single
mutants. Moreover the 100I + 103N double mutant,
which is completely resistant to efavirenz, was strongly
inhibited (EC50 ) 7.95 nΜ). Compared to compound 5,
the acrylonitrile derivative 6b (Z-isomer) showed slightly
increased potency against single and double mutants
but was clearly less active than the corresponding
E-isomer 6a.
Differences of potency between 6a (E) and 6b (Z) were
in agreement with the docking studies, which have
suggested stronger interactions of the cyano group in
E-orientation with the W229 region in the binding site
of the RT enzyme but were less favorable for the
compound with the Z-orientation.
With the introduction of a methyl group in the R or â
position to the cyano substituent in compound 6a,
activity against mutant strains was not much affected
except for the increased potency on the 100I +103N
[EC50 ) 7.95 nΜ (6c) and 2.51 nΜ (6f)] double mutant
strain. Upon comparison of 6c (E-isomer) with 6d (Z-
isomer), the data indicated a somewhat lower activity
level of the Z-isomer compared to the E-isomer, which
was somewhat more outspoken in the 188L, 103 + 181,
and 100I + 103N mutant strains with this modification.
Introduction of an acetonitrile group (compound 6e)
instead of a methyl group (compound 6d) had a decreas-
ing effect on activity against the panel of mutant strains
but on the other hand was more potent against LAI
virus (compound 6e).
a Conditions: (a) SOCl2, CH2Cl2, room temperature, 9 h; (b)
NaCN, DMF, 80 °C, 9 h; (c) 3-hydroxypropionitrile 25, 80 °C, 9 h.
by reaction with the 1,1′-carbonyldiimidazole (CDI) in
tetrahydrofuran (THF).
Alkyl or alkyloxy chains have been introduced as
spacer groups according to the synthesis pathway
described in Scheme 6. Compounds 7e and 7f were
easily synthesized from the chlorine derivative 24
derived from methyl alcohol 13.
Biological Results and Discussion
All compounds were tested for potency (EC50, nano-
molar), indicating concentrations required to achieve
50% protection of MT-4 cells from HIV-1 cytopathicity
according to the MTT method.18 The compounds were
tested against a panel consisting of wild-type LAI virus
(IIIB), single (103N, 181C, 188L, 100I, and 227C) and
double mutant (103 + 181, 100I + 103N, 227L + 106A)
strains derived from it. Cytotoxicity (CC50) was also
determined to assess selectivity of the antiviral effect.
Table 1 lists EC50 and CC50 values for the compounds
of formula 6 and 7. The ratio between the CC50 and EC50
values, which is called the selectivity index (SI), indi-
cates the specificity of the antiviral effect.
Table 1 shows the results of our SAR strategy.
Introduction of the G spacer groups into derivative 5
led to very potent compounds demonstrating high
The replacement of the methyl group in compound
6f by an electron-withdrawing group such as a chlorine
Table 1. Cytotoxicity and Inhibition of HIV-1
EC50 (nM)
103N +
100I +
227L +
cmpd
5
G-CN
E/Z LAI
CC50
SIa
103N
181C
6.3
188L
100I
227C
181C
103N
106A
-CN
0.4
100/0 0.5
0/100 0.6
100/0 0.8
0/100 0.8
5000 12 500
30 000 60 000
30 000 50 000
20 000 25 000
3000 3750
2000 5012
30 000 30 000
10 000 10 000
30 000 7500
3000 4782
1000 1578
10 000 3162
10 000 12 589
2
7.9
2
31
3.16
31.6
31.6
4
20
7.95
20
50
31.6
251
31
100
2
4
2.51
5
12.5
5
40
31
1
1000
12.6
1
6a -CHdCH-CN
6b -CHdCH-CN
6c -CHdC(Me)-CN
6d -CHdC(Me)-CN
0.3
1.6
0.6
1
1.26
5
1.6
4
6.31
2.51
31.6
6.31
12.5
2
40
25.1
31.6
2
0.4
6.3
0.6
7.95
794
31.6
39.8
2
4
4
3.16
20
>10 000
5
6e -CHdC(CH2-CN)-CN 12/88 0.4
6.31
1
2.51
2
0.4
0.63
7.95
7.95
3.16
31.6
0.8
40
ndb
nd
25.1
251
31.6
31.6
12.5
500
nd
nd
25.1
5
6f
6h -CHdCH-CH2-CN
6i -C(Cl)dCH-CN
-C(Me)dCH-CN
100/0
100/0
70/30
1
2.51
1
6.31
2.51
5
1
50
7.95
4
4
2.51 31.6
7.95
1
31.6
40
25.1
40
316
2510
7a -CH2-CH2-CN
7c -CH2-CH(Me)-CN
7d -2-furyl-5-CN
7e -CH2-CN
7f
2
0.63
0.63
4
4
6.31
31.6
10
10
160
nd
nd
3.16
31.6
25.1
125
25.1
nd
nd
0.8
nd
nd
-CH2-O-(CH2)2-CN
efavirenz
nevirapin
delavirdine
0.63 10 000 15 873
1
39.8
40
160
10 000 10 000
20 000 >500 6310
40
40
>10 000
nd
1
3
10 000
2000
>10 000
1260
>10 000
20 000
63.1 >20 000 >300 2510
nd
nd
a SI ) CC50/EC50 b nd, not determined.
.

2076 Journal of Medicinal Chemistry, 2005, Vol. 48, No. 6
Guillemont et al.
Chart 1. Comparison of Antiviral Activity between
Compound 6a and Efavirenz
Conclusion
This study clearly demonstrated that introduction of
new spacer groups G on DAPY compounds 6 and 7 was
crucial to generate very potent derivatives against the
HIV-1 wild-type and a number of clinically relevant
single and double mutants. The integrated structure-
activity relationship strategy has led to the identifica-
tion of 6a,c,f and 7c. These derivatives have displayed
an excellent in vitro potency with better antiviral
activity against HIV-1 resistant strains in comparison
with other currently approved NNRTIs tested. Several
members of this class of compounds are being considered
as promising candidates for clinical evaluation.
a pEC50 ) -log EC50
.
Experimental Section
atom (6h) resulted in a somewhat reduced potency. To
evaluate the influence of the chain length, a homologous
compound 6g derived from 6a was prepared and this
modification resulted in a reduced activity against the
panel of mutant strains.
When a 2-furyl heterocycle (7d) was inserted as G
spacer group instead of the double bond into compound
5, this also resulted in reduced activity.
The reduction of the double bond in 6a and 6c led to
very attractive compounds in terms of activity (7a and
7c). The potency against single and double mutant
strains was not affected by the presence of a methyl
group on the side chain. Nevertheless, these two com-
pounds were slightly less active than the corresponding
vinylcyanide derivatives 6a and 6c, especially on 188L
and 100I + 103N. Omission of one carbon in the side
chain (7e) or the incorporation of an alkyloxy group (7f)
caused a decrease of the potency against the mutant
strains when compared to 6a.
Structural analogues of 5, therefore, have demon-
strated high activity as inhibitors of HIV-1 reverse
transcriptase. The trend of the potency is
-CHdCH-CN (E) 6a > -CHdC(Me)-CN (E) 6c >
-C(Me)dCH-CN (E) 6f > -CH2-CH2-CN 7a.
Derivative 6a was found to be extremely potent not
only against the wild-type virus (EC50 ) 0.5 nΜ) but
moreover against a panel of 15 clinically relevant
mutant strains (Chart 1).
Considering activity against single or double mutant
strain in this panel, the DAPY derivative 6a had a
better overall profile compared to efavirenz. The gain
in activity was substantial, especially on the double
mutant 100I + 103N (>1000 times more active than
efavirenz). In DAPY compounds, like TMC125 and
compound 6a, the number of mutations generally plays
a more important role in resistance than a single
particular mutation, and up to now no pattern could be
found, notwithstanding that the 181C mutation is part
of the picture. Compound 6a has been assessed against
a series of HIV-1 strains harboring one or two RT-
associated mutations (obtained by site-directed muta-
genesis), which are considered resistant to currently
approved NNRTIs. From the available data, only the
179F + 181C mutant strain showed a >10-fold increase
in EC50 for 6a. This combination of mutations was
observed upon in vitro selection experiments with
TMC125,19 and hence cross-resistance between the two
compounds is not altogether unexpected. It should be
noted that, in the case of NNRTIs, in vitro results rarely
are predictive for the clinical situation.
General. All analytically pure compounds were dried under
vacuum in a drying pistol by use of a Buchi Glass Oven B-580
apparatus. Melting points were determined on a Leica VMHB
apparatus and are uncorrected. TLC analyses were run on
silica gel 60 F254 plates (Merck) with a variety of solvent
systems and a fluorescent indicator for visualization. Spots
were visualized under 254 nm UV illumination. Column
chromatography was performed with silica gel 60 (Merck)
(0.015-0.040 mm) or Kromasyl (Akzo Nobel) (0.010 mm).
Proton NMR spectra were recorded on Bruker Avance 300 (300
MHz) and Bruker Avance 400 (400 MHz) spectrometers with
internal deuterium lock. Chemicals shifts are reported to
internal dimethyl sulfoxide (DMSO) (δ 2.54) in parts per
million (ppm) and coupling constants (J) are given in hertz
(Hz). Mass spectral data (MS) were collected on a Applied
Biosystems API100 electrospray/quadripole instrument. High-
resolution mass spectral data (HRMS) were recorded on a
Waters LCT electrospray/time-of-flight instrument. Elemental
analyses were performed on Thermo Electron Corporation
instruments EA 1110 or EA 1108 for C, H, N and the results
were within (0.4% of the theoretical values. Compound 6d
analysis showed the percent H found to be 0.5% lower than
percent H calculated. However, 1H NMR spectroscopic analysis
and TLC revealed a pure compound. Chemicals or solvents
were purchased from either Acros Co or Aldrich Chemical Co.
Yields refer to purified products and are not optimized.
4-[2-(4-Cyanophenylamino)pyrimidin-4-ylamino]-3,5-
dimethylbenzoic Acid Ethyl Ester 10. A mixture of 4-amino-
3,5-dimethylbenzoic acid ethyl ester 8 (3.48 g, 18 mmol) and
4-(4-chloropyrimidin-2-ylamino)benzonitrile 9(8) (4.15 g, 18
mmol) was stirred at 150 °C in an oil bath for 2 h. The residue
was crystallized from diisopropyl ether and CH2Cl2. Compound
10 (6.5 g, 93%) was obtained as a pale yellow solid, mp 200
1
°C; H NMR (DMSO-d6) δ 1.4 (3 H, t, J ) 7 Hz), 2.2 (6 H, s)
4.4 (2 H, q, J ) 7.0 Hz), 6.5 (1 H, br s), 7.2-7.6 (4 H, m), 7.8
(2 H, s), 8.07 (1 H, d, J ) 5.5 Hz), 10-10.6 (2 H, m). Anal.
(C22H21N5O2) C, H, N.
4-[4-(4-Bromo-2,6-dimethylphenylamino)pyrimidin-2-
ylamino]benzonitrile 12. A mixture of 4-(4-chloropyrimidin-
2-ylamino)benzonitrile 9(8) (3 g, 13 mol) and intermediate
4-bromo-2,6-dimethylphenylamine 11 (2.6 g, 13 mol) was
stirred at 150 °C for 1 h. The residue was poured into 10%
aqueous K2CO3 and extracted with 5% of CH3OH in CH2Cl2.
The combined organic layers were dried (MgSO4) and concen-
trated. The residue was crystallized from diisopropyl ether and
CH3CN to afford 12 (3.2 g, 62%) as a pale yellow solid, mp
1
221 °C; H NMR (DMSO-d6) δ 2.15 (6 H, s), 6.08 (1 H, br s),
7.38 (2 H, s), 7.45 (1 H, d, J ) 8.6 Hz), 7.77 (2 H, d, J ) 8.6
Hz), 8.02 (2 H, d, J ) 5.5 Hz), 8.56 (1 H, s), 9.3 (1 H, s). Anal.
(C19H16BrN5) C, H, N.
4-[4-(4-Hydroxymethyl-2,6-dimethylphenylamino)-
pyrimidin-2-ylamino]benzonitrile 13. Lithium aluminum
hydride (1.55 g, 41 mmol) was added portionwise at 5 °C to a
mixture of 10 (4.26 g, 11 mmol) in THF (100 mL) under
nitrogen. The mixture was stirred at 0 °C for 1 h and at room
temperature overnight. Ethyl acetate was added, and then
water, and the mixture was filtered over Celite. The organic

DAPY Analogues: Antiviral Activity against HIV-1
Journal of Medicinal Chemistry, 2005, Vol. 48, No. 6 2077
layer was separated, dried (MgSO4), and concentrated under
vacuum. The residue was crystallized from diisopropyl ether
to give 13 (3 g, 79%) as a white solid, mp 246 °C; 1H NMR
(DMSO-d6) δ 2.16 (6 H, s), 4.5 (2 H, s), 4.88 (1 H, s), 6 (1 H, br
s), 7.11 (2 H, s), 7.4-7.8 (4 H, m), 7.97 (1 H, d, J ) 5.5 Hz),
8.45 (1 H, s), 9.23 (1 H, s). HRMS: calcd for C20H20N5O (MH)+
m/z 346.1668, found 346.1663.
4-[4-(4-Formyl-2,6-dimethylphenylamino)pyrimidin-2-
ylamino]benzonitrile 14. To a solution of 13 (5.4 g, 16 mmol)
in CH2Cl2 (100 mL) was added MnO2 (5.4 g, 62 mol). The
mixture was stirred at room temperature for 20 h, filtered
through Celite, and concentrated under vacuum. The residue
was crystallized from 10% AcOEt in diisopropyl ether to give
14 (3.8 g, 69%) as a pale yellow solid, mp 228 °C; 1H NMR
(DMSO-d6) δ 2.3 (6 H, s), 6.15 (1 H, d, J ) 5.6 Hz), 7.4 (2 H,
d, J ) 8.6 Hz), 7.7-7.75 (4 H, m), 8.04 (1 H, d, J ) 5.6 Hz),
8.82 (1 H, s), 9.3 (1 H, s), 10.02 (1 H, s). Anal. (C20H17N5O) C,
H, N.
(E)- and (Z)-4-{4-[4-(2-Cyanovinyl)-2,6-dimethylphe-
nylamino]pyrimidin-2-ylamino}benzonitrile 6a and 6b:
First Method (Scheme 2). Potassium terbutoxide (0.25 g,
2.2 mmol) was added at 5 °C under nitrogen to a solution of
cyanomethyl triphenylphosphonium chloride 15a (0.74 g, 2.2
mmol) in THF (7 mL). The mixture was stirred at 5 °C for 30
min and then at room temperature for 30 min. A solution of
the aldehyde 14 (0.5 g, 1.5 mmol) in THF (7 mL) was added
dropwise, and the reaction was stirred at room temperature
for 8 h, followed by addition of H2O and extraction with AcOEt.
The organic layer was separated, dried (MgSO4), and concen-
trated. Compounds 6a and 6b were separated by silica gel
column chromatography (96:4:0.1 toluene/iPrOH/NH4OH) and
crystallized from CH3CN and Et2O.
H, s), 7.4-7.75 (6 H, m), 8.05 (1 H, d, J ) 5.5 Hz), 8.9 (1 H, s),
9.65 (1 H, s). Anal. (C23H20N6) C, H, N.
2-{4-[2-(4-Cyanophenylamino)pyrimidin-4-ylamino]-
3,5-dimethylbenzylidene}succinonitrile 6e (E/Z 12/88).
Tributylphosphine (0.44 mL, 1.75 mmol) was added at room
temperature to a solution of (2E)-but-2-enedinitrile 16a (0.14
g, 1.75 mmol) in THF (7 mL), and the mixture was refluxed
for 2 h. A solution of 14 (0.2 g, 0.58 mmol) in THF (5 mL) was
added. The mixture was refluxed overnight, followed by
addition of H2O and extraction with CH2Cl2. The combined
organic layers were dried (MgSO4) and concentrated. The
residue was silica gel column chromatographed (99:1 CH2Cl2/
CH3OH). Crystallization from diisopropyl ether and CH3CN
(50:50) gave 6e (0.036 g, 15%) as a white solid, mp 204 °C; 1H
NMR (DMSO-d6) δ 2.2 (6 H, s), 4 (2 H, s), 6.35 (1 H, br s),
7.28-7.8 (7 H, m), 8.02 (1 H, d, J ) 5.6 Hz), 8.98 (1 H, br s),
9.63 (1 H, br s). HRMS: calcd for C24H20N7 (MH)+ m/z 406.1780,
found 406.1791.
(E)-4-{4-[4-(2-Cyano-1-methylvinyl)-2,6-dimethylphe-
nylamino]pyrimidin-2-ylamino}benzonitrile 6f. A mix-
ture of 12 (0.6 g, 1.52 mmol), crotononitrile 16c (E/Z ) 33/66)
(0.87 mL, 10.68 mmol), Pd(OAc)2 (0.066 g, 0.29 mmol), Et3N
(0.216 mL, 1.52 mmol), and tri-o-tolylphosphine (0.462 g, 1.52
mmol) in CH3CN (15 mL) was stirred in a sealed vessel at
150 °C for 72 h. H2O was added and the mixture was extracted
with CH2Cl2. The combined organic layers were dried (MgSO4)
and concentrated. The residue was silica gel column chro-
matographed (98:2 CH2Cl2/CH3OH) and only the E isomer was
isolated. Crystallization from diisopropyl ether and CH3CN
(50:50) gave 6f (0.032 g, 11%) as a white powder, mp 224 °C;
1H NMR (DMSO-d6) δ 2.2 (6 H, s), 2.45 (3 H, s), 5.5-6.4 (2 H,
m), 7.3-7.8 (6 H, m), 8.02 (1 H, d, J ) 5.6 Hz), 8.92 (1 H, s),
9.6 (s, 1 H). Anal. (C23H20N6) C, H, N.
Compound 6a (0.262 g, 48%) white solid, mp 245 °C; 1H
NMR (DMSO-d6) δ 2.2 (6 H, s), 6.1 (1 H, d, J ) 5.6 Hz), 6.35
(1 H, d, J ) 16.7 Hz), 7.43 (2 H, d, J ) 7.7 Hz), 7.46 (2 H, s),
7.58 (1 H, d, J ) 16.7 Hz), 7.77 (2 H, d, J ) 7.7 Hz), 8.02 (1 H,
d, J ) 5.6 Hz), 8.6 (1 H, br s), 9.2 (1 H, br s). Anal. (C22H18N6)
C, H, N.
Compound 6b (0.127 g, 23%) as a white solid, mp 258 °C;
1H NMR (DMSO-d6) δ 2.2 (6 H, s), 5.9 (1 H, d, J ) 12.1 Hz),
6.35 (1 H, br s), 7.35-7.67 (7 H, m), 8 (1 H, d, J ) 5.6 Hz), 9.0
(1 H, br s), 9.6 (1 H, br s). HRMS: calcd for C22H19N6 (MH)+
m/z 367.1871, found 367.1682.
Second Method (Scheme 3). A mixture of 12 (0.08 g, 0.21
mmol), acrylonitrile 16b (0.14 mL, 2.13 mmol), Pd(OAc)2 (0.009
g, 0.043 mmol), Et3N (0.03 mL, 0.21 mmol), and tri-o-
tolylphosphine (0.064 g, 0.21 mmol) in CH3CN (7 mL) was
stirred in a sealed vessel at 150 °C overnight. H2O was added
and the mixture was extracted with CH2Cl2. The combined
organic layers were dried (MgSO4) and concentrated. The
residue was silica gel column chromatographed (80:20
CH2Cl2/AcOEt) to give a residue of 6a and 6b (E/Z ) 80/20).
Crystallization from CH3CN and Et2O afforded 6a (0.035 g,
55%).
(E)-4-{4-[4-(3-Cyanopropenyl)-2,6-dimethylphenyl-
amino]pyrimidin-2-ylamino}benzonitrile 6g. A mixture
of 12 (0.2 g, 0.51 mmol), allylcyanide 16d (0.41 mL, 5.1 mmol),
Pd(OAc)2 (0.023 g, 0.1 mmol), Et3N (0.14 mL, 0.1 mmol) and
tri-o-tolylphosphine (0.155 g, 0.51 mmol) in CH3CN (10 mL)
was stirred in a sealed vessel at 150 °C overnight. H2O was
added and the mixture was extracted with CH2Cl2. The
combined organic layers were dried (MgSO4) and concentrated.
The residue was silica gel column chromatographed (99:1
CH2Cl2/CH3OH). Crystallization from 5% AcOEt in diisopropyl
ether yielded 6g (0.04 g, 21%) as a white solid, mp 213 °C; 1H
NMR (DMSO-d6) δ 2.18 (6 H, s), 3.52 (2 H, dd, J ) 6.1 and 1.5
Hz) 6.0 (1 H, br s), 6.23 (1 H, dt, J ) 15.7 and 6.1 Hz), 6.7 (1
H, d, J ) 15.7 Hz), 7.25 (2 H, s), 7.45 (2 H, d, J ) 8.1 Hz), 7.8
(2 H, d, J ) 8.1 Hz), 8.0 (1 H, d, J ) 6.1 Hz), 8.5 (1 H, s), 9.3
(1 H, s). Anal. (C23H20N6) C, H, N.
4-{4-[4-(2-Cyano-1-hydroxyethyl)-2,6-dimethylphenyl-
amino]pyrimidin-2-ylamino}benzonitrile 17. n-Butyllith-
ium (1.6 M in hexane, 2.4 mL, 3.8 mmol) was added dropwise
at -70 °C to a solution of diisopropylamine (0.5 mL, 3.8 mmol)
in THF (5 mL) under nitrogen. The mixture was stirred at
-20 °C for 30 min and recooled at -70 °C before dropwise
addition of a solution of CH3CN (0.2 mL, 3.8 mmol) in THF (2
mL). The mixture was stirred at -20 °C for 1 h and cooled
again at -70 °C. A solution of 14 (0.3 g, 0.95 mmol) in THF (2
mL) was added. The mixture was stirred for 2 h, followed by
addition of H2O and extraction with AcOEt. The combined
organic layers were dried (MgSO4) and concentrated. The
residue was silica gel column chromatographed (98:2 CH2Cl2/
CH3OH). Crystallization from diisopropyl ether and CH3CN
(50:50) afforded 17 (0.18 g, 49%) as a white solid, mp 252 °C;
1H NMR (DMSO-d6) δ 2.20 (6 H, s), 2.82-3.97 (2 H, m), 4.91
(1 H, br s), 5.68 (1 H, br s), 6.0 (1 H, br s), 7.21 (2 H, s), 7.48
(2 H, d, J ) 8.0 Hz), 7.29 (2 H, d, J ) 8.0 Hz), 7.98 (1 H, d, J
) 5.5 Hz), 8.48 (1 H, s), 9.25 (1 H, s). HRMS: calcd for
C22H21N6O (MH)+ m/z 385.1777, found 385.1805.
(E)- and (Z)-4-{4-[4-(2-Cyano-2-methylvinyl)-2,6-dim-
ethylphenylamino]pyrimidin-2-ylamino}benzonitrile 6c
and 6d. Potassium terbutoxide (2.21 g, 19.6 mmol) was added
at 5 °C under nitrogen to a solution of diethyl cyanoethylphos-
phonate 15b (3.44 mL, 19.6 mL) in THF (45 mL). The mixture
was stirred at 5 °C for 30 min and then at room temperature
for 30 min. A solution of the aldehyde 14 (4.5 g, 13 mmol) in
THF (45 mL) was added dropwise, and the reaction was stirred
at room temperature for 3 h, followed by addition of H2O and
extraction with AcOEt. The organic layer was separated, dried
(MgSO4), and concentrated. Compounds 6c and 6d were
separated by silica gel column chromatography (85:15:1 cy-
clohexane/iPrOH/NH4OH) and crystallized from 5% iPrOH in
diisopropyl ether.
Compound 6c (1.23 g, 25%) white solid, mp 201 °C; 1H NMR
(DMSO-d6) δ 2.15 (3 H, s), 2.2 (6 H, s), 6.2 (1 H, br s), 7.25 (2
H, s), 7.38-7.83 (5 H, m), 8.0 (1 H, d, J ) 5.5 Hz), 8.85 (1 H,
s), 9.55 (1 H, s). Anal. (C23H20N6) C, H, N.
Compound 6d (1.56 g, 31%) white solid, mp 246 °C; 1H NMR
(DMSO-d6) δ 2.15 (3 H, s), 2.2 (6 H, s), 6.2 (1 H, br s), 7.25 (1

2078 Journal of Medicinal Chemistry, 2005, Vol. 48, No. 6
Guillemont et al.
4-{4-[4-(2-Cyanoacetyl)-2,6-dimethylphenylamino]-
pyrimidin-2-ylamino}benzonitrile 18. Jones’s reagent (3
M, 1.87 mL, 5.6 mmol) was added at 5 °C to a mixture of
intermediate 17 (1.11 g, 2.9 mmol) in 2-propanone (20 mL)
under nitrogen. The mixture was stirred at 0 °C for 2 h and
then poured out into H2O, basified with 10% aqueous NaHCO3,
and extracted with CH2Cl2. The organic layer was separated,
dried (MgSO4), and concentrated. The residue was silica gel
column chromatographed (98:2:0.1 CH2Cl2/CH3OH/NH4OH)
and the concentrated product fractions were crystallized from
the mixture was refluxed overnight. The residue was silica gel
column chromatographed (99:1 CH2Cl /CH OH 99/1). Concen-
2
3
tration of the pure fractions provided 20 (0.13 g, 6%) as a pale
yellow oil.
4-(4-{4-[5-(Hydroxyiminomethyl)furan-2-yl]-2,6-
dimethylphenylamino}pyrimidin-2-ylamino)benzoni-
trile 21. Aqueous NaOH (5 N, 2 mL) was added dropwise at
50 °C to a mixture of 20 (0.13 g, 0.3 mmol) and hydroxylamine
chloride 23 (0.03 g, 0.41 mmol) in EtOH (10 mL). The mixture
2
was stirred at 50 °C for 2 h and
/ of this mixture was
3
i
10% PrOH in diisopropyl ether to give 18 (0.056 g, 5%) as a
concentrated before addition of H2O and then extraction with
CH2Cl2. The organic layer was washed with 10% aqueous K2-
CO3, dried over MgSO4, filtered, and concentrated. Intermedi-
ate 21 (0.21 g) was obtained as a pale yellow oil.
white powder, mp 150 °C; 1H NMR (DMSO-d6) δ 2.25 (6 H, s),
4.7 (2 H, br s), 6.12 (1 H, d, J ) 5.5 Hz), 7.45 (2 H, d, J ) 8.5
Hz), 7.73 (4 H, m), 8.05 (1 H, d, J ) 5.5 Hz), 8.8 (1 H, s), 9.35
(1 H, s). HRMS: calcd for C22H19N6O (MH)+ m/z 383.1620,
found 383.1599.
4-{4-[4-(1-Chloro-2-cyanovinyl)-2,6-dimethylphenyl-
amino]pyrimidin-2-ylamino}benzonitrile 6h (E/Z 70/30).
A solution of 18 (0.2 g, 0.52 mmol) in phosphorus oxychloride
(1.5 mL) was stirred at 80 °C for 24 h. The mixture was then
poured into ice water, basified by use of solid K2CO3, and
extracted with 10% CH3OH in CH2Cl2. The residue was silica
gel column chromatographed (99:1 CH2Cl2/CH3OH). The pure
fractions were concentrated to afford 6h (0.026 g, 13%) as a
white powder; 1H NMR (DMSO-d6) δ 2.2 (6 H, s), 6.0-6.95 (2
H, m), 7.4-8.05 (7 H, m), 9.0 (1 H, s), 9.65 (1 H, br s). MS
(ESI): m/z 401 (MH)+.
5-{4-[2-(4-Cyanophenylamino)pyrimidin-4-ylamino]-
3,5-dimethylphenyl}-furan-2-carbonitrile7d. 1,1-Carbon-
yldiimidazole (0.16 g, 1.3 mmol) was added to a mixture of 21
(0.32 mmol) in THF (20 mL). The mixture was refluxed
overnight, poured out into H2O, and extracted with CH2Cl2.
The combined organic layers were dried (MgSO4) and concen-
trated. The residue was silica gel column chromatographed
(98:2 CH2Cl2/CH3OH). Crystallization from 5% AcOEt in Et2O
provided 7d (0.05 g, 38%) as white solid, mp > 260 °C; 1H NMR
(DMSO-d6) δ 2.2 (6 H, s), 6.3 (1 H, m), 7.25 (1 H, d, J ) 3.5
Hz), 7.3-7.7 (6 H, m), 7.75 (1 H, d, J ) 3.5 Hz), 8.02 (1 H, s),
8.92 (1 H, s), 9.6 (1 H, s). Anal. (C24H18N6O2) C, H, N. HRMS:
calcd for C24H19N6O (MH)+ m/z 407.1620, found 407.1630.
4-[4-(4-Chloromethyl-2,6-dimethylphenylamino)pyri-
midin-2-ylamino]benzonitrile 24. Thionyl chloride (1 mL,
14 mmol) was added dropwise at 5 °C to a solution of 13 (0.85
g, 3 mmol) in CH2Cl2 (20 mL). The mixture was stirred at room
temperature overnight and concentrated to afford 24 (1 g,
4-{4-[4-(2-Cyano-ethyl)-2,6-dimethylphenylamino]-
pyrimidin-2-ylamino}benzonitrile 7a. A solution of 6a and
6b (E/Z ) 50/50) (0.15 g, 0.41 mmol) in CH3OH (10 mL) was
hydrogenated under 3 atm of H2 for 5 h at room temperature,
with Pd/C 10% (0.07 g) as the catalyst. The catalyst was
removed by filtration through Celite and washed with CH2-
Cl2, and the filtrate was concentrated. After crystallization
from diisopropyl ether, the residue was silica gel column
chromatographed (99:1 CH2Cl2/CH3OH). Crystallization from
CH3CN and diisopropyl ether afforded 7a (0.04 g, 27%) as a
white powder, mp 208 °C; 1H NMR (DMSO-d6) δ 2.15 (6 H, s),
2.82 (2 H, t, J ) 7.7 Hz), 2.9 (2 H, t, J ) 7.7 Hz), 6.0 (1 H, br
s), 7.1 (2 H, s), 7.45 (2 H, d, J ) 8 Hz), 7.8 (2 H, d, J ) 8 Hz),
7.95 (1 H, d, J ) 6.1 Hz), 8.4 (1 H, s), 9.2 (1 H, s). Anal.
(C23H20N6) C, H, N.
1
100%) as a pale yellow oil; H NMR (DMSO-d6) δ 2.15 (6 H,
s), 4.82 (2 H, s), 6.53 (1 H, d, J ) 5.5 Hz), 7.31 (2 H, s), 7.38,
(2 H, d, J ) 8.6 Hz), 7.50 (2 H, d, J ) 8.6 Hz), 8.12 (1 H, d, J
) 5.5 Hz), 10.82 (1 H, s), 11.3 (1 H, s).
4-[4-(4-Cyanomethyl-2,6-dimethylphenylamino)pyri-
midin-2-ylamino]benzonitrile 7e. A mixture of 24 (0.36 g,
0.99 mmol) and sodium cyanide (0.097 g, 1.9 mmol) in DMF
(7 mL) was stirred at 80 °C overnight and poured out into
water. After extraction with CH2Cl2, the organic layer was
dried (MgSO4) and concentrated. The residue was silica gel
column chromatographed (60:40 CH3OH/NH4, HCO3). Crystal-
lization from CH3CN and diisopropyl ether yielded 7e (0.035
g, 10%) as a white powder, mp 210 °C; 1H NMR (DMSO-d6) δ
2.18 (6 H, s), 3.97 (2 H, s), 6.05 (1 H, br s), 7.16 (2 H, s), 7.46
(2 H, d, J ) 8.6 Hz), 7.76 (2 H, d, J ) 8.6 Hz), 8.0 (1H, d, J )
5.5 Hz), 8.57 (1 H, s), 9.28 (1 H, s). Anal. (C21H18N6) C, H, N.
4-{4-[4-(2-Cyano-2-methylethyl)-2,6-dimethylphenyl-
amino]pyrimidin-2-ylamino}benzonitrile7b. This com-
pound was prepared from 6c and 6d (E/Z ) 50/50) (0.35 g,
0.92 mmol) as described for 7a. Compound 7b (0.147 g, 42%)
1
was obtained as a white solid, mp 187 °C; H NMR (DMSO-
d6) δ 1.3 (3 H, m), 2.13 (6 H, s), 2.8 (2 H, m), 3.2 (1 H, br s),
6.3 (1 H, s), 7.1 (2 H, s), 7.35-8.1 (5 H, m), 8.7 (1 H, m), 9.6 (1
H, m). Anal. (C23H22N6) C, H, N.
4-{4-[4-(2-Cyanoethoxymethyl)-2,6-dimethylphenyl-
amino]pyrimidin-2-ylamino}benzonitrile 7f. A solution of
24 (0.2 g, 5.5 mmol) in 3-hydroxypropionitrile 25 (2 mL) was
stirred at 80 °C overnight, poured out into H2O, and extracted
with CH2Cl2. The combined organic layer were dried (MgSO4)
and concentrated. The residue was silica gel column chro-
matographed (99:1:0.1 CH2Cl2/CH3OH/NH4OH). The residue
was washed with 3 N HCl, extracted with CH2Cl2, dried
(MgSO4), and concentrated. After crystallization from 5%
AcOEt in diisopropyl ether, 7f (0.039 g, 18%) was obtained as
4-{4-[4-(2-Cyano-1-methylethyl)-2,6-dimethylphenyl-
amino]pyrimidin-2-ylamino}benzonitrile 7c. A solution of
6f (0.065 g, 0.171 mmol) in CH3OH (5 mL) and THF (5 mL)
was hydrogenated under 3 atm of H2 for 48 h at room
temperature, with Pd/C 10% (0.065 g) as the catalyst. The
catalyst was removed by filtration through Celite and washed
with CH2Cl2, and the filtrate was concentrated. The residue
was silica gel column chromatographed (99:1 CH2Cl2/CH3OH)
1
a white solid, mp > 260 °C; H NMR (DMSO-d6) δ 2.17 (6 H,
1
to give 7c (0.035 g, 54%) as a white powder, mp 208 °C; H
s), 2.82 (2 H, t, J ) 6.1 Hz), 3.71 (2 H, t, J ) 6.1 Hz), 4.54 (2
H, s), 6.0 (1 H, br s), 7.15 (2 H, s), 7.43-7.82 (4 H, m), 7.98
(1H, d, J ) 6 Hz), 8.61 (1 H, s), 9.38 (1 H, s). Anal. (C23H22N6O)
C, H, N.
NMR (DMSO-d6) δ 1.38 (3 Η, d, J ) 7 Hz), 2.18 (6 H, s), 2.81
(2 H, d, J ) 7 Hz), 3.15 (1 H, st, J ) 7 Hz), 6.0 (1 H, br s), 7.12
(2 H, s), 7.45 (2 H, d, J ) 8 Hz), 7.80 (2 H, d, J ) 8 Hz), 7.98
(1 H, d, J ) 5.5 Hz), 8.46 (1 H, s), 9.28 (1 H, s). Anal. (C23H22N6)
C, H, N.
Test Method. The antiviral activity of compounds was
determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltet-
razolium bromide (MTT) colorimetric assay method.17 Different
concentrations of the test compounds were added to wells of a
flat-bottom microtiter plate. Then, virus and MT-4 cells were
introduced to a final concentration of 200-250 50% cell culture
infectious doses (CCID50)/well and 30,000 cells/well, respec-
tively. Cytotoxicity of the test compound was determined in
parallel by use of mock-infected cell cultures containing an
identical compound concentration range but no virus. After 5
4-{4-[4-(5-Formylfuran-2-yl)-2,6-dimethylphenylamino]-
pyrimidin-2-ylamino}benzonitrile 20. A mixture of 12 (2
g, 5.1 mmol), bis(pinacolato)diboron 22 (1.4 g, 5.5 mmol),
tetrakis(triphenylphosphine) palladium(0) (0.29 g, 0.3 mmol),
and K2CO3 (2.8 g, 20 mmol) in toluene (100 mL), EtOH (5 mL),
and H2O (10 mL) was refluxed for a weekend. 5-Bromofuran-
2-carbaldehyde 19 (0.98 g, 5.5 mmol) and K2CO3 (1.4 g, 10
mmol) were sequentially added to the previous solution and

DAPY Analogues: Antiviral Activity against HIV-1
Journal of Medicinal Chemistry, 2005, Vol. 48, No. 6 2079
(9) Gazzard, B. G.; Pozniak, A. L.; Rosenbaum, W.; Yeni, G. P.;
Staszewski, S.; Arasteh, K.; De Dier, K.; Peeters, M.; Woodfall,
B.; Stebbing, J.; Vant’ Klooster, G. A. An open-label assessment
of TMC 125-a new, next-generation NNRTI, for 7 days in HIV-1
infected individuals with NNRTI resistance. AIDS (London,
England) 2003, 17 (18), F49-54.
days of incubation at 37 °C with 5% CO2, the viability of the
HIV and mock-infected cells was assessed by the MTT
method.17 This method allows the calculation of both the 50%
effective concentration for inhibition of viral cytopathicity
(EC50) and the 50% cytotoxic concentration (CC50).
(10) Ludovici, D. W.; De Corte, B. L.; Kukla, M. J.; Ye, H.; Ho, C. Y.;
Lichenstein, M. A.; Kavash, R. W.; Andries, K.; De Bethune, M.
P.; Azijn, H.; Pauwels, R.; Lewi, P. J.; Heeres, J.; Koymans, L.
M. H.; De Jonge, M. R.; Van Aken, K. J. A.; Daeyaert, F. F. D.;
Das, K.; Arnold, E.; Janssen, P. A. J. Evolution of Anti-HIV Drug
Candidates. Part 3: Diarylpyrimidine (DAPY) Analogues. Bioorg.
Med. Chem. Lett. 2001, 11, 2235-2239.
(11) Lewi, P. J.; De Jonge, M. R.; Daeyaert, F. F. D.; Koymans, L.
M. H.; Vinkers, M. H.; Heeres, J.; Janssen, P. A. J.; Arnold, E.;
Das, K.; Clark, A. D., Jr.; Hughes, S. H.; Boyer, P. L.; De
Be´thune, M.-P.; Pauwels, R.; Andries, K.; Kukla, M.; Ludovici,
D.; De Corte, B.; Kavash, R. W.; Ho, C. Y. On the detection of
multiple-binding modes of ligands to proteins, from biological,
structural, and modeling data. J. Comput.-Aided Mol. Des. 2003,
17, 129-134.
Acknowledgment. We are grateful to Sandrine
Jolly and Sebastien Thomas for purification of all
products and Sophie Merillon for all elemental analyses.
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