Polyanion inhibitors of human immunodeficiency virus and other viruses. 5. Telomerized anionic surfactants derived from amino acids

Article abstract of DOI:10.1021/jm960493b

ω-Acryloyl anionic surfactants, whose polar heads are derived from amino acids, have been telomerized to prepare polyanions of a predetermined molecular weight. The main goal of this study was to verify whether the antiviral activity is influenced by the degree of polymerization of the polyanions. The oligomeric polyanions were evaluated for their activity against human immunodeficiency virus (HIV-1 or HIV-2) and various other RNA and DNA viruses. With regard to their anti-HIV activity, a minimum number of anionic groups was necessary to achieve an inhibitory effect. Moreover, to be active the overall conformation of the polyanion must be such that the anionic groups are located on the external site of the molecule. With some of the polyanions, a 50% inhibition concentration (IC₅₀) as low as 1 μg/mL, or even 0.1 μg/mL, was noted against HIV-1 in CEM-4 and MT-4 cells, respectively. The most potent polyanions also proved active against human cytomegalovirus and herpex simplex virus at concentrations of 5-10 and 20-40 μg/mL, respectively. No activity was observed against any of the other viruses tested (i.e., vesicular stomatitis, Sindbis, Semliki forest, parainfluenza, Junin, Tacaribe, Coxsackie, polio, reo, and vaccinia). No toxicity for the host cells was observed at concentrations up to 200 μg/mL.

Full text of DOI:10.1021/jm960493b

342
J . Med. Chem. 1997, 40, 342-349
P olya n ion In h ibitor s of Hu m a n Im m u n od eficien cy Vir u s a n d Oth er Vir u ses. 5.
Telom er ized An ion ic Su r fa cta n ts Der ived fr om Am in o Acid s†
Alain Leydet,‡ Ve´ronique Barragan,‡ Bernard Boyer,‡ J ean Louis Monte´ro,‡ J ean Pierre Roque,*,‡
Myriam Witvrouw,§ J ose´ Este,§ Robert Snoeck,§ Graciela Andrei,§ and Erick De Clercq§
Laboratoire de Chimie Organique Physique, Universite´ de Montpellier II, Place E. Bataillon,
F34095 Montpellier Ce´dex 5, France, and Rega Institute for Medical Research, Katholieke Universiteit Leuven,
10, Minderbroedersstraat, B-3000 Leuven, Belgium
Received J une 27, 1996X
ω-Acryloyl anionic surfactants, whose polar heads are derived from amino acids, have been
telomerized to prepare polyanions of a predetermined molecular weight. The main goal of
this study was to verify whether the antiviral activity is influenced by the degree of
polymerization of the polyanions. The oligomeric polyanions were evaluated for their activity
against human immunodeficiency virus (HIV-1 or HIV-2) and various other RNA and DNA
viruses. With regard to their anti-HIV activity, a minimum number of anionic groups was
necessary to achieve an inhibitory effect. Moreover, to be active the overall conformation of
the polyanion must be such that the anionic groups are located on the external site of the
molecule. With some of the polyanions, a 50% inhibition concentration (IC50) as low as 1 µg/
mL, or even 0.1 µg/mL, was noted against HIV-1 in CEM-4 and MT-4 cells, respectively. The
most potent polyanions also proved active against human cytomegalovirus and herpex simplex
virus at concentrations of 5-10 and 20-40 µg/mL, respectively. No activity was observed
against any of the other viruses tested (i.e., vesicular stomatitis, Sindbis, Semliki forest,
parainfluenza, J unin, Tacaribe, Coxsackie, polio, reo, and vaccinia). No toxicity for the host
cells was observed at concentrations up to 200 µg/mL.
Sch em e 1
In tr od u ction
The inhibition of the binding of the human immuno-
deficiency virus (HIV) envelope glycoprotein to the CD4
receptor by polyanionic substances is now well estab-
lished.1 These compounds may offer an alternative
treatment strategy as their mode of action differs from
that of the nucleoside analogues which are used for the
treatment of prevention of AIDS. Polyanions can be
useful as vaginal microbicides2 and for topical applica-
tions against respiratory tract virus infection.3 Clinical
trials are under way with some of the polyanionic
compounds.4
In a previous article,5 we have shown that polyanions,
obtained by γ-polymerization in micellar solution of
ω-unsaturated anionic surfactants derived from the 11-
undecenoic acid (Scheme 1), are inhibitory to HIV,
without toxicity to the host cells (CEM-4 or MT-4). Anti-
HIV activities of these polyanions have been demon-
strated in vitro based on a reduction of the reverse
transcriptase activity in the culture supernatant or
inhibition of the virus-induced cytopathic effect, the 50%
inhibition concentration (IC50) being in the range of 0.1-
3.5 µg/mL.
Despite the interesting inhibitory effects observed in
vitro, the in vivo use of these compounds seems to be
compromised because of their poor bioavailability due
to their high molecular weight. Indeed, polymers have
the tendency to accumulate in tissues or organs, par-
ticularly when they have a high molecular weight.6,7 To
have access to a better bioavailability, we thought to
synthesize smaller similar polyanions of a molecular
weight lower than 5000 Da, a value more prone to
transfer through biological barriers. Indeed there is
evidence that polyanions of limited molecular weight,
such as the modified cyclodextrin sulfates, can cross
biological barriers and are orally available.8,9
For this purpose we decided to use various 11-(N-
acryloyl-N-alkylamino)undecanoic acid derivatives as
starting monomeric units. Indeed those monomers can
be submitted to a telomerization process. A telomer-
ization reaction differs from a polymerization reaction
by the presence in the reaction medium of a chain
transfer agent (telogen) whose role is to limit the length
of the macromolecular chain and to permit to obtain
polymers with low degrees of polymerization (DPn).
These oligomers are called telomers.10,11
We present here the synthesis and the telomerization
of several methyl esters derived from various 11-(N-
acryloyl-N-alkylamino)undecanoic acids, used as mono-
mers. The saponification of the obtained telomers led
to small polyanions whose antiviral activities are also
reported.
Mon om er Syn th esis
Only the methyl 11-(N-acryloylamino)undecanoate (1)
has been described in the literature and used for an IR
study of the amide bands.12 We have prepared this
compound from the commercially available 11-aminoun-
decanoic acid (Scheme 2).
† This work is a part of the thesis of V. Barragan.
‡ Universite´ de Montpellier II.
N-Alkyl analogues of 1 were prepared from the 11-
bromoundecanoic acid on which the bromine has been
substituted by a methylamino or an ethylamino group,
§ Katholieke Universiteit Leuven.
X Abstract published in Advance ACS Abstracts, J anuary 1, 1997.
S0022-2623(96)00493-1 CCC: $14.00 © 1997 American Chemical Society

Polyanion Inhibitors of HIV and Other Viruses. 5
J ournal of Medicinal Chemistry, 1997, Vol. 40, No. 3 343
Sch em e 2
4) to prepare the N-ethylamino derivatives. Amino
acids were condensed as their methyl esters with methyl
11-(N-acryloyl-N-ethylamino)undecanoate by the dicy-
clohexylcarbodiimide/hydroxybenzotriazole couple.19,20
As direct coupling led to medium yields for the
previous N-ethyl derivatives, we tried to improve them
by using method B to prepare monomers derived from
the 11-aminoundecanoic acid. This method required
protection of the amine function of the 11-aminoundec-
anoic acid by a tert-butyloxycarbonyl (Boc) group.21-24
The N-protected acid25 was coupled to the amino acid
esters with the DCC/HOBt couple; then the acrylamide
function was introduced after deprotection by trifluo-
roacetic acid (Scheme 5). Actually obtained yields were
roughly in the same range as those with method A.
Sch em e 3
The structures of the monomers having an amino acid
residue as a polar head were determined without
ambiguity by the usual spectrometric methods. 1H
NMR spectroscopy (Scheme 6) revealed that the reso-
nance of the protons Ha, Hb, and Hc of the acrylamide
group appears as an ABC system.26 The proton Ha
shifted to the weak field because of the proximity of the
carbonyl group. Due to the rigidity of the acrylamide
group, protons Hb and Hc are nonequivalent and differ-
ently coupled from the proton Ha, and the observed
signals are split doublets.
Sch em e 4
Mon om er Telom er iza tion
Former monomers were telomerized so as to obtain
compounds with a controlled molecular weight and with
a maximum value near 5000 Da. The telomerization
reaction required a polymerizable monomer called a
taxogen and a transfer agent, most often a thiol, called
a telogen in the presence of a radical initiator.
by refluxing it with an aqueous solution of the corre-
sponding amine (Scheme 3). The amino acids obtained
after recrystallization in water are consistent with those
described in the literature.13 The carboxylic function
of the preceding acids was then protected by a methyl
ester function. The esterification has been realized by
reacting methanol in the presence of thionyl chlo-
ride.14,15 Methyl esters are amorphous white solids,
isolated in quantitative yields.
Monomers 2 and 3 were obtained by condensing at 0
°C the acryloyl chloride with methyl 11-(N-alkylamino)-
undecanoate in methylene chloride in the presence of
4-(dimethylamino)pyridine (DMAP) and triethylamine
used as catalyst.16,17 After purification by chromatog-
raphy on silica gel, the structure of the obtained acrylic
monomers was characterized by the usual methods. As
the anionic head, we decided to graft an amino acid unit.
We used glycine, â-alanine, and glutamic acid, as these
amino acids gave the best results in the case of the
polyanions obtained by γ-polymerization.18
All telomerizations were done with dodecanethiol as
the telogen agent. The radical initiator used, azobis-
(isobutyronitrile) (AIBN), has a first-order decomposi-
tion reaction,27 so it is progressively added to maintain
the radical concentration. Moreover, a weak amount
of AIBN is sufficient to initiate the reaction28 when the
telogen is an alkanethiol. The DPn of the telomers can
be adjusted to a desired value, depending upon the thiol/
monomer initial concentration ratio (Ro). Theoretically,
if the transfer constant is equal to unity, the DPn should
be equal to Ro.
Telomerization reactions were performed under ni-
trogen by refluxing in acetonitrile, used as solvent (80
°C), a mixture of monomer and dodecanethiol in fixed
concentrations. When the temperature was stabilized,
half of the AIBN was added at time t ) 0, the other
half being added 1 h after. At the end of the reaction,
the solvent was eliminated under reduced pressure and
Two synthesis strategies have been involved according
to the starting monomer. We used method A (Scheme
Sch em e 5

344 J ournal of Medicinal Chemistry, 1997, Vol. 40, No. 3
Leydet et al.
Ta ble 3. Telomerization of monomers 10-12a
Sch em e 6
104[M]o 105[T]o
DPn
monomers
telomers
(M)
(M)
Ro found
10, n ) 1
10T3
10T7
10T14
11T2
11T3
11T6
11T8
12T2
12T3
12T5
12T15
1.53
1.53
1.53
2.35
1.47
1.47
1.47
2.47
1.46
1.14
1.94
5.11
2.19
1.04
7.84
2.10
0.98
0.74
8.09
2.08
0.76
0.55
3
7
15
3
2.8
7.4
14
2.3
3.5
6.5
8
2.2
3.2
5.5
Sch em e 7
R ) H
(glycine)
11, n ) 2
R ) H
7
(â-alanine)
15
20
3
7
15
35
12, n ) 1
R ) (CH2)2CO2Me
(glutamic acid)
15
a
[M]o and [T]o: initial monomer and dodecanethiol concentra-
tions, respectively. Ro ) [M]o/[T]o.
Ta ble 1. Telomerization of Monomers 1-3a
in the case of the non-N-alkylated monomers. For the
N-methyl or N-ethyl monomers, the DPn values are
systematically lower than the Ro ones. This is obviously
the result of a steric effect.
Telomers as ester forms were saponified by 2 N
aqueous sodium hydroxide in excess under sonication;
the pH was then adjusted to 8.5-9 by an ion exchange
resin. Telomers as sodium salts were isolated by
lyophilization.
102[M]o
(M)
103[T]o
(M)
DPn
found
monomers
telomers
Ro
1, R ) H
1T4
1T6
1T11
1T20
2T5
2.48
2.48
2.48
2.48
0.470
0.410
1.77
1.30
4.0
4.93
3.09
2.06
1.21
0.470
0.273
0.707
0.367
8.0
5
4.3
6
11.2
20
5.3
9.5
12.7
20.3
4
8
12
20
10
15
25
35
5
2, R ) Me
3, R ) Et
2T9
Biologica l Resu lts
2T13
2T20
3T4
3T11
3T16
3T40
The tests performed in CEM-4 cells (Table 4) show
that about one-third of the telomeric polyanions exhibit
an inhibitory effect against HIV-1. At first glance, this
result is surprising since previous polyanions obtained
by polymerization of ω-unsaturated anionic surfactants
have systematically proven to be active against HIV.5,18
We think that this different behavior results from
structural differences between the two types of polyan-
ions (Scheme 8). Two factors differentiate the two
compound families, namely: (i) the presence of amide
groups close to the polymeric chain in the case of
telomers (Scheme 8a) and (ii) the number of lateral
chains bearing anionic groups.
4.0
4.0
4.0
2.0
0.80
0.33
20
50
120
10.8
16
40
a
[M]o and [T]o: initial monomer and dodecanethiol concentra-
tions, respectively. Ro ) [M]o/[T]o.
Ta ble 2. Telomerization of Monomers 4-6a
104[M]o 105[T]o
DPn
found
monomers
telomers
(M)
(M)
Ro
Concerning the former factor, it is well known that a
small structural modification (i.e., changing a methyl
versus an ethyl29 or an N-H group versus an N-ethyl
group30) can drastically alter the self-assembling prop-
erties of a surfactant. In a similar way the introduction
of a single perfluorinated residue at the end of the
polymeric chain of a telomerized tenside modifies its
micellar properties by changing the hydrophilic/lipo-
philic balance (HLB) of the compound.31 So it seems
possible that the introduction of amide groups, by
decreasing the hydrophobic character near the poly-
meric chain, disturbs the conformation of some acrylic
polyanions: the globular structure of micellar type could
not be achieved if the HLB value is inadequate. It is
noteworthy that among the three first series of polyan-
ions (1T, 2T, and 3T), in which the substituent of the
nitrogen corresponds respectively to a hydrogen, a
methyl, or an ethyl group, only the polyanions of the
third series are active. In this last case, the presence
of ethyl groups may confer a sufficient hydrophobic
character in the vicinity of the polymeric chain, to
restore a conformation close to that in a micellar
aggregate.
4, n ) 1
4T3
4T5
4T15
5T5
5T8
5T30
5T50
6T3
6T10
6T17
6T25
1.41
1.41
1.73
1.22
1.45
1.90
6.79
1.82
1.82
1.82
11.3
2.82
1.41
0.69
1.74
0.97
0.50
0.57
3.63
1.50
0.91
1.03
5
2.7
5.4
15.3
5.5
8.3
29.7
R ) H
10
25
7
15
38
(glycine)
5, n ) 2
R ) H
(â-alanine)
120 >50
6, n ) 1
R ) (CH2)2CO2Me
(glutamic acid)
5
12
20
3
10
17
110 >25
a
[M]o and [T]o: initial monomer and dodecanethiol concentra-
tions, respectively. Ro ) [M]o/[T]o.
the reaction medium was purified by gel permeation
chromatography on a Sephadex LH20 column (CH2Cl2/
CH3OH, 1/1).
1
The DPn value was determined by H NMR, compar-
ing the integration of the methyl ester protons (Hx) with
that of the terminal methyl group of the dodecanethiol
(Hy): DPn ) Hx/Hy. For each telomer, reagent concen-
trations as well as Ro and observed DPn values are
reported in Tables 1-3. It appears that the DPn values
observed are close to the corresponding Ro values only

Polyanion Inhibitors of HIV and Other Viruses. 5
J ournal of Medicinal Chemistry, 1997, Vol. 40, No. 3 345
Ta ble 4. Anti-HIV Activity of the Telomers in CEM-4 Cellsa
Ta ble 5. Anti-HIV Activity of the Telomers in MT-4 Cells
b
IC50 (µg/mL)
IC50c (µg/mL)
syncitium formation
cytopathicity
a
CC50
compd
(µg/mL)
HIV-1
HIV-2
HIV-1
HIV-2
1T6
1T11
1T20
2T5
3T16
5T5
6T25
11T2
11T6
136
>250
>250
128
20
55
5
28
>16
13
0.1
6
47
69
1.3
70
>29
30
7
27
12
50
150
30
150
30
30
6
30
10
150
150
50
150
50
30
30
6
10
30
>250
>250
>250
>250
3
a
50% cytotoxic concentration, or compound concentration re-
quired to reduce the viability of mock-infected MT-4 cells by 50%.
b
50% inhibitory concentration, or compound concentration re-
quired to inhibit HIV-induced cytopathicity in MT-4 cells by 50%.
c 50% inhibitory concentration, or concentration required to inhibit
by 50% syncitium formation between MOLT-4 cells and HIV-1
(IIIB) or HIV-2 (ROD)-infected HUT-78 cells. All data represent
mean values of two separate experiments.
Some telomer polyanions were evaluated against
HIV-1 and HIV-2 in MT-4 cells (Table 5). They were
all found to be active against the virus-induced cyto-
pathic effect and syncitium formation, albeit at rather
high IC50 values. The best activities were seen with
compound 6T25, which inhibited the cytopathic effect of
HIV-1 at an IC50 of 0.1 µg/mL, and compound 1T20,
which inhibited the cytopathic effect of HIV-2 at an IC50
of 1.3 µg/mL. These two polyanions were nontoxic (CC50
> 250 µg/mL).
The activity of this last group of polyanions was also
determined against several other RNA or DNA viruses.
Their 50% inhibitory concentrations (IC50) and the
cytotoxicities as measured in different cell lines are
reported in Tables 6 and 7, respectively. Marked
antiviral effects were noted with some of the compounds
(i.e., 3T16 and 6T25) against HCMV (IC50 in the range
of 5-10 µg/mL) and with these and two other com-
pounds (i.e., 11T2 and 11T6) against HSV (IC50 in the
range of 20-40 µg/mL). No appreciable activity was
noted with any of the compounds against various other
viruses such as vaccinia, vesicular stomatitis, parain-
fluenza-3, reo-1, Sindbis, Semliki forest, J unin, Tacar-
ibe, Coxsackie B4, and polio-1 (Table 6).
a
All data represent the average value of at least two separate
b
experiments. 50% cytotoxic concentration, or compound concen-
tration required to reduce the viability of uninfected cells by 50%
after 5 days of incubation in the presence of the compound. c 50%
inhibitory concentration, or compound concentration required to
inhibit HIV-induced cytopathicity by 50% (based on the MTT
assay). NA ) not active.
The second parameter to take into account is the
polymerization degree (DPn) of the telomers. This term
governs the number of anionic groups. In the case of
vinylic polyanions (Scheme 8b), the DPn ranked between
30 and 60.32 With telomer type polyanions, the DPn
value has been modulated between 3 and 40. From
Table 4, we note that small polyanions (DPn < 10) are
generally inactive or weakly active. However, with DPn
values higher than 10, it is difficult to obtain a correla-
tion between activity and the number of anionic groups.
Indeed, the three most active compounds (6T25, 4T15,
and 3T11) possess respectively 50, 15, and 11 anionic
groups.
Con clu sion
The telomerization reaction intended to prepare a new
series of polyanions derived from amphiphilic monomers
permitted access to low molecular weight polymers, with
the possibility to vary their degree of polymerization.
To achieve activity against HIV-1 and HIV-2, the
number of anionic groups carried by the polyanion must
be higher than 10. However a suitable HLB of the
Sch em e 8

346 J ournal of Medicinal Chemistry, 1997, Vol. 40, No. 3
Leydet et al.
Ta ble 6. Inhibitory Effect of Several Telomers on the Replication of DNA and RNA Viruses
a
IC50 (µg/mL)
cell
line
dextran
sulfate
virus
1T6
1T11
1T20
2T5
3T16
5T5
70
20
150
50
20
>200
>200
>200
>200
>200
>200
>50
>50
>200
>200
6T25
11T2
11T6
HSV-1 (KOS)
HSV-2 (G)
E6SM
E6SM
E6SM
HEL
HEL
E6SM
E6SM
Vero
Vero
Vero
Vero
Vero
Vero
HeLa
HeLa
2
2
2
0.6
0.4
>200
>200
>200
>50
>100
>100
>100
>50
>100
20
>200
70
>40
20
20
>40
40
10
5
>40
>40
>10
>10
>10
>10
>50
>50
>200
>200
40
20
70
30
23
20
20
20
25
20
HSV-1 TK- (B2006)
HCMV (AD169)
HCMV (Davis)
vaccinia
>100
>200
>200
40
15
>50
9
9
35
>50
>50
100
2
>400
>400
40
>200
>200
>200
>200
>200
>200
>50
>50
>200
>200
>100
>100
>100
>100
>100
>100
>50
>100
>100
>200
>200
>200
>200
>50
>50
>200
>200
>200
>200
>200
>200
>200
>200
>50
>40
>40
>200
>200
>200
>200
20
>20
>200
>200
>200
>200
>200
>200
>200
>200
50
43
>200
>200
>200
>200
>200
>200
>200
>200
43
50
>200
>200
vesicular stomatitis
parainfluenza-3
reovirus-1
Sindbis
Semliki forest
J unin
Tacaribe
Coxsackie B4
polio-1
>400
8
7
>50
>200
>200
>50
>200
>200
>400
>400
a
50% inhibitory concentration, or compound concentration required to reduce virus-induced cytopathicity by 50%. Virus was added in
the presence of the compounds, and the cells were further incubated until the cytopathic effect (CPE) was scored.
Ta ble 7. Cytotoxicity of the Telomers for Different Cell Lines
MMCa (µg/mL)
cell
line
dextran
sulfate
1T6
1T11
1T20
2T5
3T16
5T5
6T25
11T2
11T6
E6SM
HEL
HeLa
Vero
>400
>200
>200
>200
>200
>50
>200
>200
>200
>50
>200
>100
200
>50
>200
>200
>400
>50
>200
>200
100
>50
>200
>200
>400
>50
>200
>200
100
>50
>200
>40
400
>50
>200
>200
>200
>50
>200
>200
a
Minimum cytotoxic concentration, or compound concentration required to cause a microscopically detectable alteration of normal cell
morphology. For HEL cells, the MCC corresponds to the 50% inhibitory concentration required to inhibit cell growth by 50%.
dissolved in 5 mL of CH2Cl2 was added dropwise. Stirring was
maintained for 3 h at 0 °C; then 100 mL of CH2Cl2 was added,
and the organic phase was successively washed twice with 50
mL of aqueous 1 N HCl, twice with 50 mL of aqueous 5%
NaHCO3, and finally with H2O. After usual workup of the
organic phase and recrystallization (ether/hexane, 1/1, v/v), 6.5
g of white powder was isolated for ester 1: yield 97%; mp 55
°C; Rf 0.73 (AcOEt/EtOH, 8/2, v/v); IR (KBr, ν, cm-1) 3282 (N-
H), 2926 (Csp2-H), 1731 (CdO), 1550 (N-H, δ), 1230-1173
compound seems indispensable to lead to an overall
conformation that makes of the anionic groups exposed
at the external part of the molecule, as is the case in a
micellar structure.
Exp er im en ta l P r otocols
Merck silica gel 60 F254 (0.25 mm) plates were employed
for analytical TLC. Compounds were revealed by UV (254
nm), iodine, 20% (in water) sulfuric acid, or 10% (in methanol)
ninhydrin sprayings.
1
(C-O); H NMR (250 MHz, CDCl3) δ 1.1-1.3 (m, 12H, H4-
H9), 1.4-1.6 (m, 4H, H3, H10), 2.25 (t, 2H, H2), 3.25 (q, 2H,
Merck silica gel 60H was used for fast silica gel column
chromatography. Melting points were determined using a
Bucchi 530 apparatus. Sonication reactions were performed
with a 72434 Bioblock Scientific Vibra cell apparatus. Infrared
spectra were obtained on a IR-FT Bonem MB-100 spectrom-
eter. 1H and 13C NMR were recorded on Bruker AC200 and
WP200SY spectrometers, respectively. For 1H and 13C NMR
we have used numbers reported in Scheme 6. Chemical shifts
are expressed in ppm (δ). Mass spectra were recorded on a
J eol DX 100 spectrometer. Used matrix was m-nitrobenzylic
alcohol (NOBA) or thioglycerol (GT). Microanalyses were
performed in the analytical department of the CNRS (ENSCM-
Montpellier). C, H, and N elemental analysis was done for
most of the monomers; the observed deviations to the indicated
formula were always less than 0.4%.
Meth yl 11-(N-Acr yloyla m in o)u n d eca n oa te (1). To a
suspension of 11-aminoundecanoic acid (5.0 g, 24.9 mM, 1
equiv) in 50 mL of methanol was added thionyl chloride (2.9
mL, 40 mM, 1.6 equiv). The mixture was magnetically stirred
at room temperature for 2 h and then refluxed for 2 h. The
solvent (methanol) was evaporated under reduced pressure,
and the white residue obtained was dissolved in anhydrous
ether. After filtration and drying, the ester (4.8 g) was isolated
as a white powder (yield 89%, mp 155-156 °C, Rf 0.45 (CHCl3/
EtOH, 8/2, v/v)).
H
11), 3.6 (s, 3H, CO2Me), 5.55 (dd, 1H, Hb), 5.65 (m, 1H exch,
NH), 6.0 (dd, 1H, Hc), 6.2 (dd, 1H, Ha); 13C NMR (50.32 MHz,
CDCl3) δ 25 (C3, C10), 27 (C4, C9), 30 (C5-C8), 34 (C2), 40 (C11),
51 (CH3, ester), 126 (Cb), 131 (Ca), 165 (CdO, acrylate), 174
(CdO, ester); MS (FAB+, NOBA) m/ z 270 [M + H]+, 539 [2M
+ H]+, 238 [M + H - MeOH]+. Anal. (C15H27NO3) C, H, N.
Met h yl 11-(N-Acr yloyl-N-m et h yla m in o)u n d eca n oa t e
(2). To an aqueous solution of 40% methylamine (66 mL, 740
mM, 20 equiv) was added 11-bromoundecanoic acid (10 g, 37
mM, 1 equiv). The mixture was magnetically stirred for 7 h
at room temperature. The methylamine excess was eliminated
under reduced pressure. The obtained residue was recrystal-
lized in H2O; 4.37 g of white powder was isolated (yield 55%,
mp 136-137 °C, Rf 0.58 (CHCl3/EtOH, 1/1, v/v).
Esterification in CH3OH by thionyl chloride led to the
corresponding methyl ester (yield 96%, mp 136 °C, Rf 0.55
(CHCl3/EtOH, 8/2, v/v).
Following the procedure described for the ester 1, methyl
11-(N-methylamino)undecanoate (4 g, 17.2 mM, 1 equiv),
DMAP (100 mg), and acryloyl chloride (1.95 mL, 22.7 mM, 1.3
equiv) were used. After a short silica gel column (CH2Cl2/
EtOH, 95/5), 4.05 g of yellow oil was isolated for 2: yield 82%;
Rf 0.64 (CHCl3/EtOH, 95/5, v/v); IR (CHCl3, ν, cm-1) 2928-
1
2851 (C-H), 1735 (CdO); H NMR (250 MHz, CDCl3) δ 1.1-
1.3 (m, 12H, H4-H9), 1.4-1.6 (m, 4H, H3, H10), 2.2 (t, 2H, H2),
3.0 (2s, 3H, N-CH3), 3.3 (dt, 2H, H11), 3.6 (s, 3H, CO2Me), 5.6
(dd, 1H, Hb), 6.3 (m, 1H, Hc), 6.6 (dd, 1H, Ha); 13C NMR (50.32
MHz, CDCl3) δ 25 (C3, C10), 27 (C4, C9), 30 (C5-C8), 34 (C2), 36
(C1′), 47 (C11), 51 (CH3, ester), 127 (Cb), 128 (Ca), 166 (CdO,
To a solution of methyl 11-aminoundecanoate ester (3 g, 12
mM, 1 equiv) of 50 mL of anhydrous CH2Cl2 at 0 °C were
successively added 4-(N,N-dimethylamino)pyridine (DMAP; 90
mg) and triethylamine (3.6 mL, 26.4 mM, 2.2 equiv). The
freshly distilled acryloyl chloride (1.25 mL, 15.6 mM, 1.3 equiv)

Polyanion Inhibitors of HIV and Other Viruses. 5
J ournal of Medicinal Chemistry, 1997, Vol. 40, No. 3 347
acrylate), 175 (CdO, ester); MS (FAB+, NOBA) m/ z 284 [M +
H]+, 252 [M - MeOH]+, 228. Anal. (C16H29NO3) C, H, N.
Meth yl 11-(N-Acr yloyl-N-eth yla m in o)u n d eca n oa te (3).
According to the procedure described for the synthesis of 2,
methyl 11-(N-ethylamino)undecanoate (5.5 g, 22.6 mM, 1
equiv), DMAP (70 mg), 100 mL of CH2Cl2, and acryloyl chloride
(6 mL, 29.3 mM, 2.2 equiv) were used; 5.8 g of a yellow oil
was isolated for the ester 3: yield 89%; Rf 0.69 (CHCl3/EtOH,
95/5, v/v); IR (CHCl3, ν, cm-1) 2931-2856 (C-H), 1729 (CdO,
(C3, C10), 27.5 (C4, C9), 29 (C5-C8), 30 (C2′′, C3′′), 36 (C2), 41
and 42 (C1′), 46 and 47 (C11), 52 (2 CH3, esters), 126 and 127
(Ca, Cb), 166 (CdO, acrylate), 170 (CdO, amino acid), 173 and
174 (CdO, esters); MS (FAB+, NOBA) m/ z 441 [M + H]+, 266,
225, 176, 112. Anal. (C23H40N2O6), C, H, N.
Met h yl N-(11-(N′-(ter t-Bu t yloxyca r b on yl)a m in o)u n -
d eca n oyl)a m in o Acid Ester s 7-9. 11-(N-(ter t-Bu tyloxy-
ca r bon yl)a m in o)u n d eca n oic Acid . To a solution of tri-
ethylamine (0.9 mL, 6.55 mM, 1.5 equiv) of Boc-ON (1.18 g,
4.8 mM, 1.1 equiv) in 16 mL of a H2O/dioxane (50/50) mixture
was added 11-aminoundecanoic acid (1 g, 4.37 mM, 1 equiv).
Magnetic stirring was maintained for 3 h at room temperature;
then 20 mL of H2O and 25 mL of AcOEt were added. The
aqueous phase was acidified by adding aqueous 5% citric acid
till precipitation. After extraction with ethyl acetate, the
organic phase was concentrated under reduced pressure. After
recrystallization (hexane), 1.04 g of acid was obtained as a
white powder (yield 80%, mp 77-78 °C (lit.19 mp 79 °C), Rf
0.68 (AcOEt)).
1
ester), 1644 (CdO, amide), 1250-1170 (C-O); H NMR (250
MHz, CDCl3) δ 1.1 (m, 3H, H2′), 1.2-1.4 (m, 12H, H4-H9), 1.5-
1.6 (m, 4H, H3, H10), 2.3 (t, 2H, H2), 3.4 (m, 4H, H11, H1′), 3.7
(s, 3H, CO2Me), 5.7 (dd, 1H, Hb), 6.3 (dd, 1H, Hc), 6.6 (dd, 1H,
Ha) (J ac ) 16.7 Hz, J ab ) 8 Hz, J bc ) 2.3 Hz); 13C NMR (50.32
MHz, CDCl3) δ 13.5 and 15 (C2′), 25 (C3, C10), 27 (C4, C9), 30
(C5-C8), 34 (C2), 41 and 42.5 (C1′), 46 and 47.5 (C11), 51 (CH3,
ester), 126 (Cb), 131 (Ca), 166 (CdO, acrylate), 175 (CdO,
ester); MS (FAB+, GT) m/ z 298 [M + H]+, 406 [M + H + GT]+,
595 [2M + H]+, 242. Anal. (C17H31NO3) C, H, N.
Gen er a l Meth od . To a solution of methyl amino ester
hydrochloride (10 mM) in 40 mL of anhydrous THF at 0 °C
was successively added N-ethylmorpholine (10 mM) followed
by 11-(N-(tert-butyloxycarbonyl)amino)undecanoic acid (10
mM), HOBt (10 mM), and dicyclohexylcarbodiimide (10 mM).
The mixture was magnetically stirred for 1 h at 0 °C and then
for 18 h at room temperature. The solvent was eliminated
under reduced pressure and substituted by 50 mL of AcOEt.
The DCU was eliminated by filtration on Celite, and the
organic phase was successively washed twice with 25 mL of
saturated aqueous NaHCO3, twice with 25 mL of aqueous 10%
citric acid solution, and finally twice with 25 mL of H2O. The
compound was purified by chromatography under reduced
pressure (CH2Cl2).
Meth ylN-(11-(N ′-Acr yloyl-N′-eth yla m in o)u n d eca n oyl)-
a m in o Acid Ester . Gen er a l Meth od . To a suspension of
methyl amino ester hydrochloride (10 mM) in 50 mL of CH2-
Cl2 at 0 °C were successively added N-ethylmorpholine (1.27
mL, 10 mM, 1 equiv), hydroxybenzotriazole (1.35 g, 10 mM, 1
equiv), and 11-(N-acryloyl-N-ethylamino)undecanoic acid (2.84
g, 10 mM, 1 equiv) obtained by saponification under sonication
of the monomer 3. Then dicyclohexylcarbodiimide (2.06 g, 10
mM, 1 equiv) was fractionally added with a catalytic amount
of DMAP (100 mg). Magnetic stirring was maintained for 2 h
at 0 °C and then for 6 h at room temperature. The DCU
precipitate was eliminated by filtration on Celite, and the
volume of the reaction medium was diluted with 50 mL of CH2-
Cl2. The organic phase was successively washed twice with
50 mL of aqueous 1 N HCl, twice with 50 mL of saturated
aqueous NaHCO3, and finally twice with 50 mL of H2O. After
elimination of CH2Cl2 under reduced pressure, the residue was
purified by silica gel column chromatography (CH2Cl2/EtOH,
95/5, v/v).
Met h yl N-(11-(N′-(ter t-b u t yloxyca r b on yl)a m in o)u n -
d eca n oyl)glycin a te (7): yield 55%; mp 74-75 °C; Rf 0.56
(CHCl3/EtOH, 95/5, v/v); IR (KBr, ν, cm-1) 3264 (N-H), 2922-
2848 (C-H), 1761-1679 (CdO), 1521 (N-H, δ), 1415-1370
1
(C-H, tBu); H NMR (250 MHz, CDCl3) δ 1.2 (m, 12H, H4-
H9), 1.4 (m, 9H, tBu), 1.5 (m, 4H, H10, H3), 2.15 (t, 2H, H2), 3.0
(m, 2H, H11), 3.5 (s, 3H, CO2Me), 4.0 (d, 2H, H2′′), 4.4 (m, 1H,
NH′), 5.9 (m, 1H, NH); 13C NMR (50.32 MHz, CDCl3) δ 25 and
25.5 (C3, C10), 26.5 (C4, C9), 28.5-30.5 (C5-C8), 30 (tBu), 34.5
(C2), 36 (C2′′), 41 (C11), 52.5 (CH3, ester), 80 (C2′), 171 (CdO,
ester), 174 (CdO, amino acid); MS (FAB+, NOBA) m/ z 373
[M + H]+, 273, 90, 57 [tBu]+. Anal. (C19H36N2O5) C, H, N.
Met h yl N-(11-(N′-(ter t-b u t yloxyca r b on yl)a m in o)u n -
d eca n oyl)-â-a la n in a te (8): yield 50%; mp 62-63 °C; Rf 0.58
(CHCl3/EtOH, 95/5, v/v); IR (KBr, ν, cm-1) 3333 (N-H), 2921-
2855 (C-H), 1741-1678-1645 (CdO), 1420-1365 (C-H, tBu);
1H NMR (250 MHz, CDCl3) δ 1.2 (m, 12H, H4-H9), 1.4 (m,
9H, tBu), 1.5-1.6 (m, 4H, H10, H3), 2.1 (t, 2H, H2), 2.4 (t, 2H,
H2′′), 3.0 (m, 2H, H11), 3.6 (s, 3H, CO2Me), 3.45 (m, 2H, H3′′),
4.6 (m, 1H, NH′), 6.1 (m, 1H, NH); MS (FAB+, NOBA) m/ z
387 [M + H]+, 410 [M + Na + H]+, 287, 57 [tBu]+. Anal.
(C20H38N2O5), C, H, N.
Meth yl N-(11-(N′-a cr yloyl-N′-eth yla m in o)u n d eca n oyl)-
glycin a te (4): yield 57%; mp 29-30 °C; Rf 0.88 (CHCl3/EtOH,
8/2, v/v); IR (KBr, ν, cm-1) 3300 (N-H), 2922-2848 (Csp3-
H), 1742 (CdO), 1648 (CdC), 1557 (N-H); 1H NMR (250 MHz,
CDCl3) δ 1.1 (m, 3H, H2′), 1.15-1.3 (m, 12H, H4-H9), 1.5-1.7
(m, 4H, H3, H10), 2.2 (t, 2H, H2), 3.1-3.4 (m, 4H, H11, H1′), 3.65
(s, 3H, CO2Me), 3.9 (d, 2H, H2′′), 5.6 (dd, 1H, Hb) 6.3 (dd, 1H,
Hc), 6.45 (m, 1H, NH), 6.5 (dd, 1H, Ha); 13C NMR (50.32 MHz,
CDCl3) δ 13 and 14.5 (C2′), 25 (C3, C10), 27 (C4, C9), 30 (C5-
C8), 34.5 (C2), 36 (C2′′), 42 and 43 (C1′), 46 and 47 (C11), 51.5
(CH3, ester), 128 (Ca and Cb), 166 (CdO, acrylate), 170 (CdO,
ester), 174 (CdO, amino acid); MS (FAB+, GT) m/ z 355 [M +
H]+, 463 [M + GT + H]+, 210, 112. Anal. (C19H34N2O4) C, H,
N.
Meth yl N-(11-(N′-a cr yloyl-N′-eth yla m in o)u n d eca n oyl)-
â-a la n in a te (5): yield 52%; Rf 0.85 (CHCl3/EtOH, 8/2, v/v);
IR (CHCl3, ν, cm-1) 3303 (N-H), 2925-2851 (C-H), 1741
(CdO), 1641 (CdC), 1544 (N-H); 1H NMR (250 MHz, CDCl3)
δ 1.1 (m, 3H, H2′), 1.15-1.3 (m, 12H, H4-H9), 1.4-1.65 (m,
4H, H3, H10), 2.1 (t, 2H, H2), 2.6 (t, 2H, H2′′), 3.2-3.4 (m, 4H,
H1′, H11), 3.45 (m, 2H, H3′′), 3.6 (s, 3H, CO2Me), 5.6 (dd, 1H,
Hc), 6.1 (dd, 1H, NH), 6.2 (dd, 1H, Hb), 6.4 (dd, 1H, Ha); 13C
NMR (50.32 MHz, CDCl3) δ 12.5-14.5 (C2′), 25 (C3, C10), 26
(C4, C9), 30 (C5-C8), 34 (C3′′), 35 (C2), 36.5 (C2′′), 41 and 42 (C1′),
46 and 47 (C11), 51 (CO2Me), 126 and 127 (Ca, Cb), 166 (CdO,
acrylate), 170 (CdO, amino acid), 174 (CdO, ester); MS (FAB+,
GT) m/ z 369 [M + H]+, 477 [M + H + GT]+, 845 [2M + H +
GT]+. Anal. (C20H36N2O4) C, H, N.
Met h yl N-(11-(N′-(ter t-b u t yloxyca r b on yl)a m in o)u n -
d eca n oyl)-L-glu ta m a te (9): yield 54.5%; mp 64-65 °C; Rf
0.61 (CHCl3/EtOH, 95/5, v/v); 1H NMR (250 MHz, CDCl3) δ
1.1 (m, 12H, H4-H9), 1.4 (m, 9H, tBu), 1.6 (m, 4H, H10, H3),
1.9 (m, 2H, H3′′), 2.1 (t, 2H, H2), 2.4 (m, 2H, H2′′), 3.0 (m, 2H,
H11
), 3.6 and 3.7 (2s, 6H, 2CO2Me), 4.5 (m, 1H, H4′′), 4.6 (m,
1H, NH), 6.1 (m, 1H, NH); MS (FAB
+, NOBA) m/ z 459 [M +
H]+, 449, 302, 202. Anal. (C23H42N2O7) C, H, N.
Met h yl N-(11-(N′-Acr yloyla m in o)u n d eca n oyl)a m in o
Acid Ester s 10-12. Gen er a l Meth od . The preceding Boc-
amino esters (5 mM) were dissolved in 15 mL of a mixture
composed of trifluoroacetic acid and CH2Cl2 (1/1, v/v). After
stirring for 1 h at room temperature, the reaction mixture was
evaporated under reduced pressure. The residue was tritu-
rated several times with anhydrous ether; the trifluoroacetate
salts of the corresponding methyl N-(11-(N′-acryloylamino)-
undecanoyl)amino esters were isolated as a white powder in
a roughly quantitative yield.
Meth yl N-(11-(N′-a cr yloyl-N′-eth yla m in o)u n d eca n oyl)-
L-glu ta m a te (6): yield 49%; Rf 0.80 (CHCl3/EtOH, 8/2, v/v);
IR (CH3Cl, ν, cm-1) 3428 (N-H), 2931-2856 (C-H), 1736
1
(CdO, ester), 1663-1630 (CdO, amide); H NMR (250 MHz,
CDCl3) δ 1.1 (m, 3H, H2′), 1.15-1.3 (m, 12H, H4-H9), 1.4-1.6
(m, 4H, H3, H10), 2.0 (m, 2H, H3′′), 2.2 (t, 2H, H2), 2.4 (m, 2H,
H2′′), 3.2-3.5 (m, 4H, H1′, H11), 3.7 and 3.75 (2s, 6H, 2CO2Me),
4.6 (m, 1H, H4′′), 5.6 (dd, 1H, Hb), 6.4 (m, 1H, NH), 6.45 (dd,
In 50 mL of CH2Cl2 at 0 °C were added the trifluoroacetate
of methyl N-(11-aminoundecanoyl)amino ester (2 mM, 1 equiv),
triethylamine (0.61 mL, 4.4 mM, 2.2 equiv), and a catalytic
1H, Hc), 6.5 (dd, 1H, Ha) (J ac ) 16.7 Hz, J ab ) 12.5 Hz, J bc
)
2.28 Hz); 13C NMR (50.32 MHz, CDCl3) δ 12.5-14.5 (C2′), 26

348 J ournal of Medicinal Chemistry, 1997, Vol. 40, No. 3
Leydet et al.
amount of DMAP; then acryloyl chloride (0.21 mL, 2.6 mM,
1.3 equiv) dissolved in 5 mL of CH2Cl2 was added dropwise.
The reaction mixture was maintained under magnetic stirring
for 3 h at 0 °C. After addition of 50 mL of CH2Cl2, the organic
phase was successively washed twice with 50 mL of CH2Cl2.
The organic phase was successively washed twice with 50 mL
of aqueous 1 N HCl, twice with 50 mL of H2O, twice with 50
mL of aqueous 5% NaHCO3, and finally with H2O. After
evaporation under reduced pressure, the obtained residue was
recrystallized (ether).
2 (HIV-2, strain LAV-2ROD), herpex simplex virus type 1 (HSV-
1, strain KOS), thymidine kinase deficient (TK-) HSV-1 (strain
B2006), herpes simplex virus type 2 (HSV-2, strain G), human
cytomegalovirus (HCMV, strains AD169 and Davis), vaccina
virus, vesicular stomatitis virus, parainfluenza virus type 3,
reovirus type 1, J unin virus, Tacaribe virus, Sindbis virus,
Semliki forest virus, Coxsackie B4 virus, and poliovirus type
1, has been described previously.33,34
Cytotoxicity measurements were based on either microscopic
examination of alteration of normal cell morphology or inhibi-
tion of cell growth. The cell lines used for both the antiviral
activity and cytotoxicity assays were CEM-4, MT-4, Vero,
HeLa, human embryonic lung (HEL), and human embryonic
skin-muscle (E6SM) fibroblasts.
Meth yl N-(11-(N′-a cr yloyla m in o)u n d eca n oyl)glycin a te
(10): yield 47%; mp 61-62 °C; Rf 0.73 (AcOEt); IR (CHCl3, ν,
cm-1) 3298 (N-H), 2923-2848 (Csp3-H), 1742 (CdO, ester),
1
1640 (CdO, amide); H NMR (250 MHz, CDCl3) δ 1.15-1.35
(m, 12H, H4-H9), 1.4-1.65 (m, 4H, H3, H10), 2.15 (t, 2H, H2),
3.2 (q, 2H, H11), 3.65 (s, 3H, CO2Me), 4.0 (d, 2H, H2′′), 5.5 (dd,
1H, Hb), 5.55 (m, 1H, NH), 6.0 (m, 1H, NH), 6.05 (dd, 1H, Hc),
6.2 (dd, 1H, Ha); 13C NMR (50.32 MHz, CDCl3) δ 25.5 (C3, C10),
27 (C4, C9), 29 (C5-C8), 36 (C2), 39 (C2′′), 41 (C11), 52 (CH3,
ester), 126 (Cb), 131 (Ca), 166 (CdO, acrylate), 171 (C1′′), 175
(C1); MS (FAB+, NOBA) m/ z 327 [M + H]+, 238, 184, 90. Anal.
(C17H30N2O4) C, H, N.
Ack n ow led gm en t. The work carried out at UMII
was supported by the Agence Nationale de Recherche
sur le SIDA (ANRS). Also V.B. is grateful to ANRS for
a grant. We thank Dr. A. Bousseau from Rhoˆne-Poulenc
Rorer for the RT assays and anti-HIV assays in CEM-4
cells. Work at the Rega Institute was supported in part
by the Biomedical Research Programme of the European
Commission, and Belgian Nationaal Fonds voor Weten-
schappelijk Onderzoek (NFWO), and the Belgian Fonds
voor Geneeskundig Wetenschappelijk Onderzoek
(FGWO). We thank Anita Camps, Frieda De Meyer,
Anja Van Cauwenberge, Anita Van Lierde, and Barbara
Van Remoortel for excellent technical assistance.
Meth yl N-(11-(N′-a cr yloyla m in o)u n d eca n oyl)-â-a la n i-
n a te (11): yield 48% mp 103 °C; Rf 0.59 (CHCl3/EtOH, 95/5,
v/v); IR (CHCl3, ν, cm-1) 3032-3018 (C-H), 1724-1709 (CdO);
1H NMR (250 MHz, CDCl3) δ 1.1-1.3 (m, 12H, H4-H9), 1.4-
1.6 (2t, 4H, H10, H3), 2.1 (t, 2H, H2), 2.5 (t, 2H, H2′′), 3.3 (q,
2H, H11), 3.45 (q, 2H, H3′′), 3.6 (s, 3H, CO2Me), 5.5 (dd, 1H,
Hb), 6.0 (m, 1H, NH), 6.1 (m, 1H, Hc), 6.2 (dd, 1H, Ha); 13C
NMR (50.32 MHz, CDCl3) δ 25 (C3, C10), 26 (C4, C9), 29.5 (C5-
C8), 34 (C2′′), 36 (C3′′), 39 (C11), 51 (CH3), 126 (Cb), 130 (Cb),
165 (CdO, acrylate), 174 (CdO, ester); MS (FAB+, GT) m/ z
341 [M + H]+, 449 [M + H + GT]+, 287. Anal. (C18H32N2O4)
C, H, N.
Refer en ces
(1) Ito, M.; Baba, M.; Pauwels, R.; De Clercq, E.; Shigeta, S.
Inhibitory effect of dextran sulfate and heparin on the replication
of human immunodeficiency virus (HIV) in vitro. Antiviral Res.
1987, 7, 361-367. Ueno, R.; Kuno, S. Dextran sulfate, a potent
anti-HIV agent in vitro having synergism with zidovudine.
Lancet 1987, 1, 1379. Baba, M.; Nakajima, M.; Schols, D.;
Pauwels, R.; Balzarini, J .; De Clercq, E. Pentosanpolysulfate, a
sulfated oligosaccharide, is a potent and selective anti-HIV agent
in vitro. Antiviral Res. 1988, 9, 335-343. Yoshida, O.; Na-
kashima, H.; Toshida, T.; Kanedo, Y.; Matsuzaki, K.; Uryu, T.;
Yamamoto, N. Sulfatation of the immunomodulating polysac-
charide Lentinan: A novel strategy for antivirals to human
immunodeficiency virus (HIV). Biochem. Pharmacol. 1988, 37,
2887-2889. Baba, M.; Schols, D.; De Clercq, E.; Pauwels, R.;
Nagy, M.; Gyo¨rgyi-Edele´nyi, J .; Lo¨w, M.; Go¨ro¨g, S. Novel sulfated
polymers as highly potent and selective inhibitors of human
immunodeficiency virus replication and giant cell formation.
Antimicrob. Agents Chemother. 1990, 34, 134-138. Cushman,
M.; Wang, P.; Chang, S. H.; Wild, C.; De Clercq, E.; Schols, D.;
Goldman, M. E.; Bowen, J . A. Preparation and anti-HIV activi-
ties of aurintricarboxylic acid fractions and analogues: direct
correlation of antiviral potency with molecular weight. J . Med.
Chem. 1991, 34, 329-337.
Meth yl N-(11-(N′-a cr yloyla m in o)u n d eca n oyl)-L-glu ta -
m a te (12): yield 47%; mp 70 °C; Rf 0.64 (CHCl3/EtOH, 90/
10, v/v); IR (CHCl3, ν, cm-1) 3511-3410 (N-H), 2931-2855
1
(C-H), 1734-1724-1716 (CdO), 1624 (CdC); H NMR (250
MHz, CDCl3) δ 1.1-1.3 (m, 12H, H4-H9), 1.45 (t, 2H, H3), 1.55
(t, 2H, H10), 1.9 (sx, 2H, H3′′), 2.15 (t, 2H, H2), 2.4 (m, 2H, H2′′),
3.2 (t, 2H, H11), 3.6 and 3.7 (2s, 6H, 2CO2Me), 4.55 (m, 1H,
H
4′′), 5.5 (dd, 1H, Hb), 6.0 (m, 1H, NH), 6.1 (m, 1H, Hc), 6.2
(dd, 1H, Ha), 6.3 (m, 1H, NH); 13C NMR (50.32 MHz, CDCl3) δ
26 (C3, C10), 27 (C4, C9), 28 (C3′′), 29 (C5-C8), 30 (C2′′), 36 (C2),
39.5 (C11), 51 (C4′′), 52 (2CH3), 125 (Cb), 131 (Ca), 166 (CdO,
amide), 174 (CdO, esters); MS (IE) m/ z 412 [M]+, 352 [M -
CO2Me]+, 238. Anal. (C21H36N2O6) C, H, N.
Telom er iza tion Rea ction . Gen er a l P r oced u r e. The
reaction was realized in acetonitrile as solvent under nitrogen.
AIBN (Aldrich) was recrystallized twice from ethanol. The
monomer was dissolved in acetonitrile (dried and distilled from
CaH2); then the telogen (dodecanethiol, previously distilled
from CaO) was added, and the reaction mixture was heated
to 80 °C before adding half of the amount of AIBN. The AIBN
remaining was added 1 h later. The reaction medium was
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